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Sample records for activity-dependent bulk endocytosis

  1. Actin- and dynamin-dependent maturation of bulk endocytosis restores neurotransmission following synaptic depletion.

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    Tam H Nguyen

    Full Text Available Bulk endocytosis contributes to the maintenance of neurotransmission at the amphibian neuromuscular junction by regenerating synaptic vesicles. How nerve terminals internalize adequate portions of the presynaptic membrane when bulk endocytosis is initiated before the end of a sustained stimulation is unknown. A maturation process, occurring at the end of the stimulation, is hypothesised to precisely restore the pools of synaptic vesicles. Using confocal time-lapse microscopy of FM1-43-labeled nerve terminals at the amphibian neuromuscular junction, we confirm that bulk endocytosis is initiated during a sustained tetanic stimulation and reveal that shortly after the end of the stimulation, nerve terminals undergo a maturation process. This includes a transient bulging of the plasma membrane, followed by the development of large intraterminal FM1-43-positive donut-like structures comprising large bulk membrane cisternae surrounded by recycling vesicles. The degree of bulging increased with stimulation frequency and the plasmalemma surface retrieved following the transient bulging correlated with the surface membrane internalized in bulk cisternae and recycling vesicles. Dyngo-4a, a potent dynamin inhibitor, did not block the initiation, but prevented the maturation of bulk endocytosis. In contrast, cytochalasin D, an inhibitor of actin polymerization, hindered both the initiation and maturation processes. Both inhibitors hampered the functional recovery of neurotransmission after synaptic depletion. Our data confirm that initiation of bulk endocytosis occurs during stimulation and demonstrates that a delayed maturation process controlled by actin and dynamin underpins the coupling between exocytosis and bulk endocytosis.

  2. Quantitative monitoring of activity-dependent bulk endocytosis of synaptic vesicle membrane by fluorescent dextran imaging

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    Clayton, Emma Louise; Cousin, Michael Alan

    2012-01-01

    Activity-dependent bulk endocytosis (ADBE) is the dominant synaptic vesicle (SV) retrieval mode in central nerve terminals during periods of intense neuronal activity. Despite this fact there are very few real time assays that report the activity of this critical SV retrieval mode. In this paper we report a simple and quantitative assay of ADBE using uptake of large flourescent dextrans as fluid phase markers. We show that almost all dextran uptake occurs in nerve terminals, using co-localisation with the fluorescent probe FM1-43. We also demonstrate that accumulated dextran cannot be unloaded by neuronal stimulation, indicating its specific loading into bulk endosomes and not SVs. Quantification of dextran uptake was achieved by using thresholding analysis to count the number of loaded nerve terminals, since monitoring the average fluorescence intensity of these nerve terminals did not accurately report the extent of ADBE. Using this analysis we showed that dextran uptake occurs very soon after stimulation and that it does not persist when stimulation terminates. Thus we have devised a simple and quantitative method to monitor ADBE in living neurones, which will be ideal for real time screening of small molecule inhibitors of this key SV retrieval mode. PMID:19766140

  3. Synaptic Vesicle Endocytosis in Different Model Systems

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    Quan Gan

    2018-06-01

    Full Text Available Neurotransmission in complex animals depends on a choir of functionally distinct synapses releasing neurotransmitters in a highly coordinated manner. During synaptic signaling, vesicles fuse with the plasma membrane to release their contents. The rate of vesicle fusion is high and can exceed the rate at which synaptic vesicles can be re-supplied by distant sources. Thus, local compensatory endocytosis is needed to replenish the synaptic vesicle pools. Over the last four decades, various experimental methods and model systems have been used to study the cellular and molecular mechanisms underlying synaptic vesicle cycle. Clathrin-mediated endocytosis is thought to be the predominant mechanism for synaptic vesicle recycling. However, recent studies suggest significant contribution from other modes of endocytosis, including fast compensatory endocytosis, activity-dependent bulk endocytosis, ultrafast endocytosis, as well as kiss-and-run. Currently, it is not clear whether a universal model of vesicle recycling exist for all types of synapses. It is possible that each synapse type employs a particular mode of endocytosis. Alternatively, multiple modes of endocytosis operate at the same synapse, and the synapse toggles between different modes depending on its activity level. Here we compile review and research articles based on well-characterized model systems: frog neuromuscular junctions, C. elegans neuromuscular junctions, Drosophila neuromuscular junctions, lamprey reticulospinal giant axons, goldfish retinal ribbon synapses, the calyx of Held, and rodent hippocampal synapses. We will compare these systems in terms of their known modes and kinetics of synaptic vesicle endocytosis, as well as the underlying molecular machineries. We will also provide the future development of this field.

  4. Signaling induced by hop/STI-1 depends on endocytosis

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    Americo, Tatiana A.; Chiarini, Luciana B.; Linden, Rafael

    2007-01-01

    The co-chaperone hop/STI-1 is a ligand of the cell surface prion protein (PrP C ), and their interaction leads to signaling and biological effects. Among these, hop/STI-1 induces proliferation of A172 glioblastoma cells, dependent on both PrP C and activation of the Erk pathway. We tested whether clathrin-mediated endocytosis affects signaling induced by hop/STI-1. Both hyperosmolarity induced by sucrose and monodansyl-cadaverine blocked Erk activity induced by hop/STI-1, without affecting the high basal Akt activity typical of A172. The endocytosis inhibitors also affected the sub-cellular distribution of phosphorylated Erk, consistent with blockade of the latter's activity. The data indicate that signaling induced by hop/STI-1 depends on endocytosis. These findings are consistent with a role of sub-cellular trafficking in signal transduction following engagement by PrP C by ligands such as hop/STI-1, and may help help unravel both the functions of the prion protein, as well as possible loss-of-function components of prion diseases

  5. Enhanced clathrin-dependent endocytosis in the absence of calnexin.

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    Hao-Dong Li

    Full Text Available Calnexin, together with calreticulin, constitute the calnexin/calreticulin cycle. Calnexin is a type I endoplasmic reticulum integral membrane protein and molecular chaperone responsible for the folding and quality control of newly-synthesized (glycoproteins. The endoplasmic reticulum luminal domain of calnexin is responsible for lectin-like activity and interaction with nascent polypeptide chains. The role of the C-terminal, cytoplasmic portion of calnexin is not clear.Using yeast two hybrid screen and immunoprecipitation techniques, we showed that the Src homology 3-domain growth factor receptor-bound 2-like (Endophilin interacting protein 1 (SGIP1, a neuronal specific regulator of endocytosis, forms complexes with the C-terminal cytoplasmic domain of calnexin. The calnexin cytoplasmic C-tail interacts with SGIP1 C-terminal domains containing the adaptor complexes medium subunit (Adap-Comp-Sub region. Calnexin-deficient cells have enhanced clathrin-dependent endocytosis in neuronal cells and mouse neuronal system. This is reversed by expression of full length calnexin or calnexin C-tail.We show that the effects of SGIP1 and calnexin C-tail on clathrin-dependent endocytosis are due to modulation of the internalization of the receptor-ligand complexes. Enhanced clathrin-dependent endocytosis in the absence of calnexin may contribute to the neurological phenotype of calnexin-deficient mice.

  6. Dynamin-dependent amino acid endocytosis activates mechanistic target of rapamycin complex 1 (mTORC1).

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    Shibutani, Shusaku; Okazaki, Hana; Iwata, Hiroyuki

    2017-11-03

    The mechanistic target of rapamycin complex 1 (mTORC1) is a master regulator of protein synthesis and potential target for modifying cellular metabolism in various conditions, including cancer and aging. mTORC1 activity is tightly regulated by the availability of extracellular amino acids, and previous studies have revealed that amino acids in the extracellular fluid are transported to the lysosomal lumen. There, amino acids induce recruitment of cytoplasmic mTORC1 to the lysosome by the Rag GTPases, followed by mTORC1 activation by the small GTPase Ras homolog enriched in brain (Rheb). However, how the extracellular amino acids reach the lysosomal lumen and activate mTORC1 remains unclear. Here, we show that amino acid uptake by dynamin-dependent endocytosis plays a critical role in mTORC1 activation. We found that mTORC1 is inactivated when endocytosis is inhibited by overexpression of a dominant-negative form of dynamin 2 or by pharmacological inhibition of dynamin or clathrin. Consistently, the recruitment of mTORC1 to the lysosome was suppressed by the dynamin inhibition. The activity and lysosomal recruitment of mTORC1 were rescued by increasing intracellular amino acids via cycloheximide exposure or by Rag overexpression, indicating that amino acid deprivation is the main cause of mTORC1 inactivation via the dynamin inhibition. We further show that endocytosis inhibition does not induce autophagy even though mTORC1 inactivation is known to strongly induce autophagy. These findings open new perspectives for the use of endocytosis inhibitors as potential agents that can effectively inhibit nutrient utilization and shut down the upstream signals that activate mTORC1. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Prominin-2 expression increases protrusions, decreases caveolae and inhibits Cdc42 dependent fluid phase endocytosis

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    Singh, Raman Deep, E-mail: Takhter.Ramandeep@mayo.edu; Schroeder, Andreas S.; Scheffer, Luana; Holicky, Eileen L.; Wheatley, Christine L.; Marks, David L., E-mail: Marks.david@mayo.edu; Pagano, Richard E.

    2013-05-10

    Highlights: •Prominin-2 expression induced protrusions that co-localized with lipid raft markers. •Prominin-2 expression decreased caveolae, caveolar endocytosis and increased pCav1. •Prominin-2 expression inhibited fluid phase endocytosis by inactivation of Cdc42. •These endocytic effects can be reversed by adding exogenous cholesterol. •Caveolin1 knockdown restored fluid phase endocytosis in Prominin2 expressing cells. -- Abstract: Background: Membrane protrusions play important roles in biological processes such as cell adhesion, wound healing, migration, and sensing of the external environment. Cell protrusions are a subtype of membrane microdomains composed of cholesterol and sphingolipids, and can be disrupted by cholesterol depletion. Prominins are pentaspan membrane proteins that bind cholesterol and localize to plasma membrane (PM) protrusions. Prominin-1 is of great interest as a marker for stem and cancer cells, while Prominin-2 (Prom2) is reportedly restricted to epithelial cells. Aim: To characterize the effects of Prom-2 expression on PM microdomain organization. Methods: Prom2-fluorescent protein was transfected in human skin fibroblasts (HSF) and Chinese hamster ovary (CHO) cells for PM raft and endocytic studies. Caveolae at PM were visualized using transmission electron microscopy. Cdc42 activation was measured and caveolin-1 knockdown was performed using siRNAs. Results: Prom2 expression in HSF and CHO cells caused extensive Prom2-positive protrusions that co-localized with lipid raft markers. Prom2 expression significantly decreased caveolae at the PM, reduced caveolar endocytosis and increased caveolin-1 phosphorylation. Prom2 expression also inhibited Cdc42-dependent fluid phase endocytosis via decreased Cdc42 activation. Effects on endocytosis were reversed by addition of cholesterol. Knockdown of caveolin-1 by siRNA restored Cdc42 dependent fluid phase endocytosis in Prom2-expressing cells. Conclusions: Prom2 protrusions primarily

  8. CD82 endocytosis and cholesterol-dependent reorganization of tetraspanin webs and lipid rafts

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    Xu, Congfeng; Zhang, Yanhui H.; Thangavel, Muthusamy; Richardson, Mekel M.; Liu, Li; Zhou, Bin; Zheng, Yi; Ostrom, Rennolds S.; Zhang, Xin A.

    2009-01-01

    Tetraspanin CD82 suppresses cell migration, tumor invasion, and tumor metastasis. To determine the mechanism by which CD82 inhibits motility, most studies have focused on the cell surface CD82, which forms tetraspanin-enriched microdomains (TEMs) with other transmembrane proteins, such as integrins. In this study, we found that CD82 undergoes endocytosis and traffics to endosomes and lysosomes. To determine the endocytic mechanism of CD82, we demonstrated that dynamin and clathrin are not essential for CD82 internalization. Depletion or sequestration of sterol in the plasma membrane markedly inhibited the endocytosis of CD82. Despite the demand on Cdc42 activity, CD82 endocytosis is distinct from macropinocytosis and the documented dynamin-independent pinocytosis. As a TEM component, CD82 reorganizes TEMs and lipid rafts by redistributing cholesterol into these membrane microdomains. CD82-containing TEMs are characterized by the cholesterol-containing microdomains in the extreme light- and intermediate-density fractions. Moreover, the endocytosis of CD82 appears to alleviate CD82-mediated inhibition of cell migration. Taken together, our studies demonstrate that lipid-dependent endocytosis drives CD82 trafficking to late endosomes and lysosomes, and CD82 reorganizes TEMs and lipid rafts through redistribution of cholesterol.—Xu, C., Zhang, Y. H., Thangavel, M., Richardson, M. M., Liu, L., Zhou, B., Zheng, Y., Ostrom, R. S., Zhang, X. A. CD82 endocytosis and cholesterol-dependent reorganization of tetraspanin webs and lipid rafts. PMID:19497983

  9. Coupling of exocytosis and endocytosis at the presynaptic active zone.

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    Maritzen, Tanja; Haucke, Volker

    2018-02-01

    Brain function depends on the ability of neurons to communicate with each other via the regulated exocytosis of neurotransmitter-containing synaptic vesicles (SVs) at specialized presynaptic release sites termed active zones (AZs). The presynaptic AZ comprises an assembly of large multidomain proteins that link the machinery for vesicle fusion to sites of voltage-dependent Ca 2+ entry. Following SV fusion at AZ release sites SV membranes are retrieved by compensatory endocytosis, and SVs are reformed. Recent data suggest that Ca 2+ -triggered SV exocytosis at AZs and endocytic retrieval of SVs may be functionally and physically linked. Here we discuss the evidence supporting such exo-endocytic coupling as well as possible modes and mechanisms that may underlie coupling of exocytosis and endocytosis at and around AZs in presynaptic nerve terminals. As components of the exo-endocytic machinery at synapses have been linked to neurological and neuropsychiatric disorders, understanding the mechanisms that couple exocytosis and endocytosis at AZs may be of importance for developing novel therapies to treat these diseases. Copyright © 2017 Elsevier Ireland Ltd and Japan Neuroscience Society. All rights reserved.

  10. CD4- and dynamin-dependent endocytosis of HIV-1 into plasmacytoid dendritic cells

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    Pritschet, Kathrin; Donhauser, Norbert; Schuster, Philipp; Ries, Moritz; Haupt, Sabrina; Kittan, Nicolai A.; Korn, Klaus [Institute of Clinical and Molecular Virology, National Reference Centre for Retroviruses, Friedrich-Alexander-Universitaet Erlangen-Nuernberg, 91054 Erlangen (Germany); Poehlmann, Stefan [Institute of Virology, Hannover Medical School, 30625 Hannover (Germany); Holland, Gudrun; Bannert, Norbert [Robert Koch-Institute, Center for Biological Security 4, 13353 Berlin (Germany); Bogner, Elke [Institute of Virology, Charite University Hospital, 10117 Berlin (Germany); Schmidt, Barbara, E-mail: baschmid@viro.med.uni-erlangen.de [Institute of Clinical and Molecular Virology, National Reference Centre for Retroviruses, Friedrich-Alexander-Universitaet Erlangen-Nuernberg, 91054 Erlangen (Germany)

    2012-02-20

    Chronic immune activation, triggered by plasmacytoid dendritic cell (PDC) interferon (IFN)-alpha production, plays an important role in HIV-1 pathogenesis. As the entry of HIV-1 seems to be important for the activation of PDC, we directly characterized the viral entry into these cells using immuno-electron microscopy, cellular fractionation, confocal imaging, and functional experiments. After attachment to PDC, viruses were taken up in an energy-dependent manner. The virions were located in compartments positive for caveolin; early endosomal antigen 1; Rab GTPases 5, 7 and 9; lysosomal-associated membrane protein 1. PDC harbored more virus in endocytic vesicles than CD4+ T cells (p < 0.05). Blocking CD4 inhibited the uptake of virions into cytosolic and endosomal compartments. Dynasore, an inhibitor of dynamin-dependent endocytosis, not the fusion inhibitor T-20, reduced the HIV-1 induced IFN-alpha production. Altogether, our morphological and functional data support the role of endocytosis for the entry and IFN-alpha induction of HIV-1 in PDC.

  11. Lysosomal enzyme delivery by ICAM-1-targeted nanocarriers bypassing glycosylation- and clathrin-dependent endocytosis.

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    Muro, Silvia; Schuchman, Edward H; Muzykantov, Vladimir R

    2006-01-01

    Enzyme replacement therapy, a state-of-the-art treatment for many lysosomal storage disorders, relies on carbohydrate-mediated binding of recombinant enzymes to receptors that mediate lysosomal delivery via clathrin-dependent endocytosis. Suboptimal glycosylation of recombinant enzymes and deficiency of clathrin-mediated endocytosis in some lysosomal enzyme-deficient cells limit delivery and efficacy of enzyme replacement therapy for lysosomal disorders. We explored a novel delivery strategy utilizing nanocarriers targeted to a glycosylation- and clathrin-independent receptor, intercellular adhesion molecule (ICAM)-1, a glycoprotein expressed on diverse cell types, up-regulated and functionally involved in inflammation, a hallmark of many lysosomal disorders. We targeted recombinant human acid sphingomyelinase (ASM), deficient in types A and B Niemann-Pick disease, to ICAM-1 by loading this enzyme to nanocarriers coated with anti-ICAM. Anti-ICAM/ASM nanocarriers, but not control ASM or ASM nanocarriers, bound to ICAM-1-positive cells (activated endothelial cells and Niemann-Pick disease patient fibroblasts) via ICAM-1, in a glycosylation-independent manner. Anti-ICAM/ASM nanocarriers entered cells via CAM-mediated endocytosis, bypassing the clathrin-dependent pathway, and trafficked to lysosomes, where delivered ASM displayed stable activity and alleviated lysosomal lipid accumulation. Therefore, lysosomal enzyme targeting using nanocarriers targeted to ICAM-1 bypasses defunct pathways and may improve the efficacy of enzyme replacement therapy for lysosomal disorders, such as Niemann-Pick disease.

  12. The function of endocytosis in Wnt signaling.

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    Brunt, Lucy; Scholpp, Steffen

    2018-03-01

    Wnt growth factors regulate one of the most important signaling networks during development, tissue homeostasis and disease. Despite the biological importance of Wnt signaling, the mechanism of endocytosis during this process is ill described. Wnt molecules can act as paracrine signals, which are secreted from the producing cells and transported through neighboring tissue to activate signaling in target cells. Endocytosis of the ligand is important at several stages of action: One central function of endocytic trafficking in the Wnt pathway occurs in the source cell. Furthermore, the β-catenin-dependent Wnt ligands require endocytosis for signal activation and to regulate gene transcription in the responding cells. Alternatively, Wnt/β-catenin-independent signaling regulates endocytosis of cell adherence plaques to control cell migration. In this comparative review, we elucidate these three fundamental interconnected functions, which together regulate cellular fate and cellular behavior. Based on established hypotheses and recent findings, we develop a revised picture for the complex function of endocytosis in the Wnt signaling network.

  13. Endophilin, Lamellipodin, and Mena cooperate to regulate F-actin-dependent EGF-receptor endocytosis.

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    Vehlow, Anne; Soong, Daniel; Vizcay-Barrena, Gema; Bodo, Cristian; Law, Ah-Lai; Perera, Upamali; Krause, Matthias

    2013-10-16

    The epidermal growth factor receptor (EGFR) plays an essential role during development and diseases including cancer. Lamellipodin (Lpd) is known to control lamellipodia protrusion by regulating actin filament elongation via Ena/VASP proteins. However, it is unknown whether this mechanism supports endocytosis of the EGFR. Here, we have identified a novel role for Lpd and Mena in clathrin-mediated endocytosis (CME) of the EGFR. We have discovered that endogenous Lpd is in a complex with the EGFR and Lpd and Mena knockdown impairs EGFR endocytosis. Conversely, overexpressing Lpd substantially increases the EGFR uptake in an F-actin-dependent manner, suggesting that F-actin polymerization is limiting for EGFR uptake. Furthermore, we found that Lpd directly interacts with endophilin, a BAR domain containing protein implicated in vesicle fission. We identified a role for endophilin in EGFR endocytosis, which is mediated by Lpd. Consistently, Lpd localizes to clathrin-coated pits (CCPs) just before vesicle scission and regulates vesicle scission. Our findings suggest a novel mechanism in which Lpd mediates EGFR endocytosis via Mena downstream of endophilin.

  14. Equine arteritis virus is delivered to an acidic compartment of host cells via clathrin-dependent endocytosis

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    Nitschke, Matthias; Korte, Thomas; Tielesch, Claudia; Ter-Avetisyan, Gohar; Tuennemann, Gisela; Cardoso, M. Cristina; Veit, Michael; Herrmann, Andreas

    2008-01-01

    Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae. Infection by EAV requires the release of the viral genome by fusion with the respective target membrane of the host cell. We have investigated the entry pathway of EAV into Baby Hamster Kindey cells (BHK). Infection of cells assessed by the plaque reduction assay was strongly inhibited by substances which interfere with clathrin-dependent endocytosis and by lysosomotropic compounds. Furthermore, infection of BHK cells was suppressed when clathrin-dependent endocytosis was inhibited by expression of antisense RNA of the clathrin-heavy chain before infection. These results strongly suggest that EAV is taken up via clathrin-dependent endocytosis and is delivered to acidic endosomal compartments

  15. Size-dependent endocytosis of gold nanoparticles studied by three-dimensional mapping of plasmonic scattering images

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    Lee Chia-Wei

    2010-12-01

    Full Text Available Abstract Background Understanding the endocytosis process of gold nanoparticles (AuNPs is important for the drug delivery and photodynamic therapy applications. The endocytosis in living cells is usually studied by fluorescent microscopy. The fluorescent labeling suffers from photobleaching. Besides, quantitative estimation of the cellular uptake is not easy. In this paper, the size-dependent endocytosis of AuNPs was investigated by using plasmonic scattering images without any labeling. Results The scattering images of AuNPs and the vesicles were mapped by using an optical sectioning microscopy with dark-field illumination. AuNPs have large optical scatterings at 550-600 nm wavelengths due to localized surface plasmon resonances. Using an enhanced contrast between yellow and blue CCD images, AuNPs can be well distinguished from cellular organelles. The tracking of AuNPs coated with aptamers for surface mucin glycoprotein shows that AuNPs attached to extracellular matrix and moved towards center of the cell. Most 75-nm-AuNPs moved to the top of cells, while many 45-nm-AuNPs entered cells through endocytosis and accumulated in endocytic vesicles. The amounts of cellular uptake decreased with the increase of particle size. Conclusions We quantitatively studied the endocytosis of AuNPs with different sizes in various cancer cells. The plasmonic scattering images confirm the size-dependent endocytosis of AuNPs. The 45-nm-AuNP is better for drug delivery due to its higher uptake rate. On the other hand, large AuNPs are immobilized on the cell membrane. They can be used to reconstruct the cell morphology.

  16. alpha-Adducin mutations increase Na/K pump activity in renal cells by affecting constitutive endocytosis: implications for tubular Na reabsorption.

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    Torielli, Lucia; Tivodar, Simona; Montella, Rosa Chiara; Iacone, Roberto; Padoani, Gloria; Tarsini, Paolo; Russo, Ornella; Sarnataro, Daniela; Strazzullo, Pasquale; Ferrari, Patrizia; Bianchi, Giuseppe; Zurzolo, Chiara

    2008-08-01

    Genetic variation in alpha-adducin cytoskeletal protein is implicated in the polymerization and bundling of actin and alteration of the Na/K pump, resulting in abnormal renal sodium transport and hypertension in Milan hypertensive rats and humans. To investigate the molecular involvement of alpha-adducin in controlling Na/K pump activity, wild-type or mutated rat and human alpha-adducin forms were, respectively, transfected into several renal cell lines. Through multiple experimental approaches (microscopy, enzymatic assays, coimmunoprecipitation), we showed that rat and human mutated forms increased Na/K pump activity and the number of pump units; moreover, both variants coimmunoprecipitate with Na/K pump. The increased Na/K pump activity was not due to changes in its basolateral localization, but to an alteration of Na/K pump residential time on the plasma membrane. Indeed, both rat and human mutated variants reduced constitutive Na/K pump endocytosis and similarly affected transferrin receptor trafficking and fluid-phase endocytosis. In fact, alpha-adducin was detected in clathrin-coated vesicles and coimmunoprecipitated with clathrin. These results indicate that adducin, besides its modulatory effects on actin cytoskeleton dynamics, might play a direct role in clathrin-dependent endocytosis. The constitutive reduction of the Na/K pump endocytic rate induced by mutated adducin variants may be relevant in Na-dependent hypertension.

  17. Activity-Dependent Ubiquitination of GluA1 Mediates a Distinct AMPAR Endocytosis and Sorting Pathway

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    Schwarz, Lindsay A.; Hall, Benjamin J.; Patrick, Gentry N.

    2010-01-01

    The accurate trafficking of AMPA receptors (AMPARs) to and from the synapse is a critical component of learning and memory in the brain, while dysfunction of AMPAR trafficking is hypothesized to be an underlying mechanism of Alzheimer’s disease. Previous work has shown that ubiquitination of integral membrane proteins is a common post-translational modification used to mediate endocytosis and endocytic sorting of surface proteins in eukaryotic cells. Here we report that mammalian AMPARs become ubiquitinated in response to their activation. Using a mutant of GluA1 that is unable to be ubiquitinated at lysines on its carboxy-terminus, we demonstrate that ubiquitination is required for internalization of surface AMPARs and their trafficking to the lysosome in response to the AMPAR agonist AMPA, but not for internalization of AMPARs in response to the NMDA receptor (NMDAR) agonist NMDA. Through over-expression or RNAi-mediated knockdown, we identify that a specific E3 ligase, Nedd4-1, is necessary for this process. Finally, we show that ubiquitination of GluA1 by Nedd4-1 becomes more prevalent as neurons mature. Together, these data show that ubiquitination of GluA1-containing AMPARs by Nedd4-1 mediates their endocytosis and trafficking to the lysosome. Furthermore, these results provide insight into how hippocampal neurons regulate AMPAR trafficking and degradation with high specificity in response to differing neuronal signaling cues, and suggest that changes to this pathway may occur as neurons mature. PMID:21148011

  18. Endocytosis and Enamel Formation

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    Cong-Dat Pham

    2017-07-01

    Full Text Available Enamel formation requires consecutive stages of development to achieve its characteristic extreme mineral hardness. Mineralization depends on the initial presence then removal of degraded enamel proteins from the matrix via endocytosis. The ameloblast membrane resides at the interface between matrix and cell. Enamel formation is controlled by ameloblasts that produce enamel in stages to build the enamel layer (secretory stage and to reach final mineralization (maturation stage. Each stage has specific functional requirements for the ameloblasts. Ameloblasts adopt different cell morphologies during each stage. Protein trafficking including the secretion and endocytosis of enamel proteins is a fundamental task in ameloblasts. The sites of internalization of enamel proteins on the ameloblast membrane are specific for every stage. In this review, an overview of endocytosis and trafficking of vesicles in ameloblasts is presented. The pathways for internalization and routing of vesicles are described. Endocytosis is proposed as a mechanism to remove debris of degraded enamel protein and to obtain feedback from the matrix on the status of the maturing enamel.

  19. Adaptor protein complex 2-mediated, clathrin-dependent endocytosis, and related gene activities, are a prominent feature during maturation stage amelogenesis.

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    Lacruz, Rodrigo S; Brookes, Steven J; Wen, Xin; Jimenez, Jaime M; Vikman, Susanna; Hu, Ping; White, Shane N; Lyngstadaas, S Petter; Okamoto, Curtis T; Smith, Charles E; Paine, Michael L

    2013-03-01

    Molecular events defining enamel matrix removal during amelogenesis are poorly understood. Early reports have suggested that adaptor proteins (AP) participate in ameloblast-mediated endocytosis. Enamel formation involves the secretory and maturation stages, with an increase in resorptive function during the latter. Here, using real-time PCR, we show that the expression of clathrin and adaptor protein subunits are upregulated in maturation stage rodent enamel organ cells. AP complex 2 (AP-2) is the most upregulated of the four distinct adaptor protein complexes. Immunolocalization confirms the presence of AP-2 and clathrin in ameloblasts, with strongest reactivity at the apical pole. These data suggest that the resorptive functions of enamel cells involve AP-2 mediated, clathrin-dependent endocytosis, thus implying the likelihood of specific membrane-bound receptor(s) of enamel matrix protein debris. The mRNA expression of other endocytosis-related gene products is also upregulated during maturation including: lysosomal-associated membrane protein 1 (Lamp1); cluster of differentiation 63 and 68 (Cd63 and Cd68); ATPase, H(+) transporting, lysosomal V0 subunit D2 (Atp6v0d2); ATPase, H(+) transporting, lysosomal V1 subunit B2 (Atp6v1b2); chloride channel, voltage-sensitive 7 (Clcn7); and cathepsin K (Ctsk). Immunohistologic data confirms the expression of a number of these proteins in maturation stage ameloblasts. The enamel of Cd63-null mice was also examined. Despite increased mRNA and protein expression in the enamel organ during maturation, the enamel of Cd63-null mice appeared normal. This may suggest inherent functional redundancies between Cd63 and related gene products, such as Lamp1 and Cd68. Ameloblast-like LS8 cells treated with the enamel matrix protein complex Emdogain showed upregulation of AP-2 and clathrin subunits, further supporting the existence of a membrane-bound receptor-regulated pathway for the endocytosis of enamel matrix proteins. These data

  20. Cyclin dependent kinase 5 regulates endocytosis in nerve terminals via dynamin I phosphorylation

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    Tan, T.C.; Hansra, G.; Calova, V.; Cousin, M.; Robinson, P.J.

    2002-01-01

    Full text: Synaptic vesicle endocytosis (SVE) in nerve terminals is essential for normal synaptic transmission and for memory retrieval. Dynamin I is a 96kDa nerve terminal phosphoprotein necessary for synaptic vesicle endocytosis in the nerve terminal. Dynamin I is dephosphorylated and rephosphorylated in a cyclical fashion with nerve terminal depolarisation and repolarisation. A number of kinases phosphorylate dynamin I in vitro including PKC, MAP kinase and cdc2. PKC phosphorylates dynamin in the proline rich domain on Ser 795 and is also thought to be the in vivo kinase for dynamin I. Another candidate is the neuron specific kinase cdk5, crucial for CNS development. The aim of this study is to identify the kinase which phosphorylates dynamin I in intact nerve terminals. Here we show that cyclin-dependent kinase 5 (cdk5) phosphorylates dynamin I in the proline-rich tail on Ser-774 or Ser-778. The phosphorylation of these sites but not Ser-795 also occurred in intact nerve terminals suggesting that cdk5 is the physiologically relevant enzyme for dynamin I. Synaptosomes prepared from rat brains (after cervical dislocations) and labelled with 32 Pi, were incubated with 100 M roscovitine (a selective inhibitor of cdks), 10 M Ro 31-8220 (a selective PKC inhibitor) and 100 M PD 98059 (a MEK kinase inhibitor). Dynamin rephosphorylation during repolarisation was reduced in synaptosomes treated with roscovitine and Ro 38-8220 but not in synaptosomes treated with PD 98059. Fluorimetric experiments on intact synaptosomes utilising FM-210 (a fluorescent dye) indicate that endocytosis was reduced in synaptosomes treated with 100 M roscovitine. Our results suggest that dynamin phosphorylation in intact nerve terminals may not be regulated by PKC or MAP kinase and that dynamin phosphorylation by cdk5 may regulate endocytosis. Copyright (2002) Australian Neuroscience Society

  1. Regulation of NKG2D-Dependent NK Cell Functions: The Yin and the Yang of Receptor Endocytosis

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    Rosa Molfetta

    2017-08-01

    Full Text Available Natural-killer receptor group 2, member D (NKG2D is a well characterized natural killer (NK cell activating receptor that recognizes several ligands poorly expressed on healthy cells but up-regulated upon stressing stimuli in the context of cancer or viral infection. Although NKG2D ligands represent danger signals that render target cells more susceptible to NK cell lysis, accumulating evidence demonstrates that persistent exposure to ligand-expressing cells causes the decrease of NKG2D surface expression leading to a functional impairment of NKG2D-dependent NK cell functions. Upon ligand binding, NKG2D is internalized from the plasma membrane and sorted to lysosomes for degradation. However, receptor endocytosis is not only a mechanism of receptor clearance from the cell surface, but is also required for the proper activation of signalling events leading to the functional program of NK cells. This review is aimed at providing a summary of current literature relevant to the molecular mechanisms leading to NKG2D down-modulation with particular emphasis given to the role of NKG2D endocytosis in both receptor degradation and signal propagation. Examples of chronic ligand-induced down-regulation of NK cell activating receptors other than NKG2D, including natural cytotoxicity receptors (NCRs, DNAX accessory molecule-1 (DNAM1 and CD16, will be also discussed.

  2. Activity-dependent ubiquitination of GluA1 mediates a distinct AMPA receptor endocytosis and sorting pathway.

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    Schwarz, Lindsay A; Hall, Benjamin J; Patrick, Gentry N

    2010-12-08

    The accurate trafficking of AMPA receptors (AMPARs) to and from the synapse is a critical component of learning and memory in the brain, whereas dysfunction of AMPAR trafficking is hypothesized to be an underlying mechanism of Alzheimer's disease. Previous work has shown that ubiquitination of integral membrane proteins is a common posttranslational modification used to mediate endocytosis and endocytic sorting of surface proteins in eukaryotic cells. Here we report that mammalian AMPARs become ubiquitinated in response to their activation. Using a mutant of GluA1 that is unable to be ubiquitinated at lysines on its C-terminus, we demonstrate that ubiquitination is required for internalization of surface AMPARs and their trafficking to the lysosome in response to the AMPAR agonist AMPA but not for internalization of AMPARs in response to the NMDA receptor agonist NMDA. Through overexpression or RNA interference-mediated knockdown, we identify that a specific E3 ligase, Nedd4-1 (neural-precursor cell-expressed developmentally downregulated gene 4-1), is necessary for this process. Finally, we show that ubiquitination of GluA1 by Nedd4-1 becomes more prevalent as neurons mature. Together, these data show that ubiquitination of GluA1-containing AMPARs by Nedd4-1 mediates their endocytosis and trafficking to the lysosome. Furthermore, these results provide insight into how hippocampal neurons regulate AMPAR trafficking and degradation with high specificity in response to differing neuronal signaling cues and suggest that changes to this pathway may occur as neurons mature.

  3. Cellular entry of G3.5 poly (amido amine) dendrimers by clathrin- and dynamin-dependent endocytosis promotes tight junctional opening in intestinal epithelia.

    Science.gov (United States)

    Goldberg, Deborah S; Ghandehari, Hamidreza; Swaan, Peter W

    2010-08-01

    This study investigates the mechanisms of G3.5 poly (amido amine) dendrimer cellular uptake, intracellular trafficking, transepithelial transport and tight junction modulation in Caco-2 cells in the context of oral drug delivery. Chemical inhibitors blocking clathrin-, caveolin- and dynamin-dependent endocytosis pathways were used to investigate the mechanisms of dendrimer cellular uptake and transport across Caco-2 cells using flow cytometry and confocal microscopy. Dendrimer cellular uptake was found to be dynamin-dependent and was reduced by both clathrin and caveolin endocytosis inhibitors, while transepithelial transport was only dependent on dynamin- and clathrin-mediated endocytosis. Dendrimers were quickly trafficked to the lysosomes after 15 min of incubation and showed increased endosomal accumulation at later time points, suggesting saturation of this pathway. Dendrimers were unable to open tight junctions in cell monolayers treated with dynasore, a selective inhibitor of dynamin, confirming that dendrimer internalization promotes tight junction modulation. G3.5 PAMAM dendrimers take advantage of several receptor-mediated endocytosis pathways for cellular entry in Caco-2 cells. Dendrimer internalization by dynamin-dependent mechanisms promotes tight junction opening, suggesting that dendrimers act on intracellular cytoskeletal proteins to modulate tight junctions, thus catalyzing their own transport via the paracellular route.

  4. The Small GTPase Rac1 Contributes to Extinction of Aversive Memories of Drug Withdrawal by Facilitating GABAA Receptor Endocytosis in the vmPFC.

    Science.gov (United States)

    Wang, Weisheng; Ju, Yun-Yue; Zhou, Qi-Xin; Tang, Jian-Xin; Li, Meng; Zhang, Lei; Kang, Shuo; Chen, Zhong-Guo; Wang, Yu-Jun; Ji, Hui; Ding, Yu-Qiang; Xu, Lin; Liu, Jing-Gen

    2017-07-26

    Extinction of aversive memories has been a major concern in neuropsychiatric disorders, such as anxiety disorders and drug addiction. However, the mechanisms underlying extinction of aversive memories are not fully understood. Here, we report that extinction of conditioned place aversion (CPA) to naloxone-precipitated opiate withdrawal in male rats activates Rho GTPase Rac1 in the ventromedial prefrontal cortex (vmPFC) in a BDNF-dependent manner, which determines GABA A receptor (GABA A R) endocytosis via triggering synaptic translocation of activity-regulated cytoskeleton-associated protein (Arc) through facilitating actin polymerization. Active Rac1 is essential and sufficient for GABA A R endocytosis and CPA extinction. Knockdown of Rac1 expression within the vmPFC of rats using Rac1-shRNA suppressed GABA A R endocytosis and CPA extinction, whereas expression of a constitutively active form of Rac1 accelerated GABA A R endocytosis and CPA extinction. The crucial role of GABA A R endocytosis in the LTP induction and CPA extinction is evinced by the findings that blockade of GABA A R endocytosis by a dynamin function-blocking peptide (Myr-P4) abolishes LTP induction and CPA extinction. Thus, the present study provides first evidence that Rac1-dependent GABA A R endocytosis plays a crucial role in extinction of aversive memories and reveals the sequence of molecular events that contribute to learning experience modulation of synaptic GABA A R endocytosis. SIGNIFICANCE STATEMENT This study reveals that Rac1-dependent GABA A R endocytosis plays a crucial role in extinction of aversive memories associated with drug withdrawal and identifies Arc as a downstream effector of Rac1 regulations of synaptic plasticity as well as learning and memory, thereby suggesting therapeutic targets to promote extinction of the unwanted memories. Copyright © 2017 the authors 0270-6474/17/377096-15$15.00/0.

  5. Endocytosis-independent function of clathrin heavy chain in the control of basal NF-κB activation.

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    Man Lyang Kim

    Full Text Available BACKGROUND: Nuclear factor-κB (NF-κB is a transcription factor that regulates the transcription of genes involved in a variety of biological processes, including innate and adaptive immunity, stress responses and cell proliferation. Constitutive or excessive NF-κB activity has been associated with inflammatory disorders and higher risk of cancer. In contrast to the mechanisms controlling inducible activation, the regulation of basal NF-κB activation is not well understood. Here we test whether clathrin heavy chain (CHC contributes to the regulation of basal NF-κB activity in epithelial cells. METHODOLOGY: Using RNA interference to reduce endogenous CHC expression, we found that CHC is required to prevent constitutive activation of NF-κB and gene expression. Immunofluorescence staining showed constitutive nuclear localization of the NF-κB subunit p65 in absence of stimulation after CHC knockdown. Elevated basal p65 nuclear localization is caused by constitutive phosphorylation and degradation of inhibitor of NF-κB alpha (IκBα through an IκB kinase α (IKKα-dependent mechanism. The role of CHC in NF-κB signaling is functionally relevant as constitutive expression of the proinflammatory chemokine interleukin-8 (IL-8, whose expression is regulated by NF-κB, was found after CHC knockdown. Disruption of clathrin-mediated endocytosis by chemical inhibition or depletion of the μ2-subunit of the endocytosis adaptor protein AP-2, and knockdown of clathrin light chain a (CHLa, failed to induce constitutive NF-κB activation and IL-8 expression, showing that CHC acts on NF-κB independently of endocytosis and CLCa. CONCLUSIONS: We conclude that CHC functions as a built-in molecular brake that ensures a tight control of basal NF-κB activation and gene expression in unstimulated cells. Furthermore, our data suggest a potential link between a defect in CHC expression and chronic inflammation disorder and cancer.

  6. Adaptor Protein Complex 2 (AP-2) Mediated, Clathrin Dependent Endocytosis, And Related Gene Activities, Are A Prominent Feature During Maturation Stage Amelogenesis

    Science.gov (United States)

    LACRUZ, Rodrigo S.; BROOKES, Steven J.; WEN, Xin; JIMENEZ, Jaime M.; VIKMAN, Susanna; HU, Ping; WHITE, Shane N.; LYNGSTADAAS, S. Petter; OKAMOTO, Curtis T.; SMITH, Charles E.; PAINE, Michael L.

    2012-01-01

    Molecular events defining enamel matrix removal during amelogenesis are poorly understood. Early reports have suggested that adaptor proteins (AP) participate in ameloblast-mediated endocytosis. Enamel formation involves the secretory and maturation stages, with an increase in resorptive function during the latter. Here, using real time PCR, we show that the expression of clathrin and adaptor protein subunits are up-regulated in maturation stage rodent enamel organ cells. AP-2 is the most up-regulated of the four distinct adaptor protein complexes. Immunolocalization confirms the presence of AP-2 and clathrin in ameloblasts with strongest reactivity at the apical pole. These data suggest that the resorptive functions of enamel cells involve AP-2 mediated, clathrin dependent endocytosis, thus implying the likelihood of a specific membrane-bound receptor(s) of enamel matrix protein debris. The mRNA expression of other endocytosis-related gene products is also up-regulated during maturation including: lysosomal-associated membrane protein 1 (Lamp1), cluster of differentiation 63 and 68 (Cd63 and Cd68), ATPase, H+ transporting, lysosomal V0 subunit D2 (Atp6v0d2), ATPase, H+ transporting, lysosomal V1 subunit B2 (Atp6v1b2), chloride channel, voltage-sensitive 7 (Clcn7) and cathepsin K (Ctsk). Immunohistological data confirms the expression of a number of these proteins in maturation stage ameloblasts. The enamel of Cd63-null mice was also examined. Despite increased mRNA and protein expression in the enamel organ during maturation, the enamel of Cd63-null mice appeared normal. This may suggest inherent functional redundancies between Cd63 and related gene products, such as Lamp1 and Cd68. Ameloblast-like LS8 cells treated with the enamel matrix protein complex Emdogain® showed up-regulation of AP-2 and clathrin subunits, further supporting the existence of a membrane-bound receptor regulated pathway for the endocytosis of enamel matrix proteins. These data together

  7. Constitutive endocytosis and turnover of the neuronal glycine transporter GlyT2 is dependent on ubiquitination of a C-terminal lysine cluster.

    Directory of Open Access Journals (Sweden)

    Jaime de Juan-Sanz

    Full Text Available Inhibitory glycinergic neurotransmission is terminated by sodium and chloride-dependent plasma membrane glycine transporters (GlyTs. The mainly glial glycine transporter GlyT1 is primarily responsible for the completion of inhibitory neurotransmission and the neuronal glycine transporter GlyT2 mediates the reuptake of the neurotransmitter that is used to refill synaptic vesicles in the terminal, a fundamental role in the physiology and pathology of glycinergic neurotransmission. Indeed, inhibitory glycinergic neurotransmission is modulated by the exocytosis and endocytosis of GlyT2. We previously reported that constitutive and Protein Kinase C (PKC-regulated endocytosis of GlyT2 is mediated by clathrin and that PKC accelerates GlyT2 endocytosis by increasing its ubiquitination. However, the role of ubiquitination in the constitutive endocytosis and turnover of this protein remains unexplored. Here, we show that ubiquitination of a C-terminus four lysine cluster of GlyT2 is required for constitutive endocytosis, sorting into the slow recycling pathway and turnover of the transporter. Ubiquitination negatively modulates the turnover of GlyT2, such that increased ubiquitination driven by PKC activation accelerates transporter degradation rate shortening its half-life while decreased ubiquitination increases transporter stability. Finally, ubiquitination of GlyT2 in neurons is highly responsive to the free pool of ubiquitin, suggesting that the deubiquitinating enzyme (DUB ubiquitin C-terminal hydrolase-L1 (UCHL1, as the major regulator of neuronal ubiquitin homeostasis, indirectly modulates the turnover of GlyT2. Our results contribute to the elucidation of the mechanisms underlying the dynamic trafficking of this important neuronal protein which has pathological relevance since mutations in the GlyT2 gene (SLC6A5 are the second most common cause of human hyperekplexia.

  8. Bovine parvovirus uses clathrin-mediated endocytosis for cell entry.

    Science.gov (United States)

    Dudleenamjil, Enkhmart; Lin, Chin-Yo; Dredge, Devin; Murray, Byron K; Robison, Richard A; Johnson, F Brent

    2010-12-01

    Entry events of bovine parvovirus (BPV) were studied. Transmission electron micrographs of infected cells showed virus particles in cytoplasmic vesicles. Chemical inhibitors that block certain aspects of the cellular machinery were employed to assess viral dependency upon those cellular processes. Chlorpromazine, ammonium chloride, chloroquine and bafilamicin A1 were used to inhibit acidification of endosomes and clathrin-associated endocytosis. Nystatin was used as an inhibitor of the caveolae pathway. Cytochalasin D and ML-7 were used to inhibit actin and myosin functions, respectively. Nocodazole and colchicine were employed to inhibit microtubule activity. Virus entry was assessed by measuring viral transcription using real-time PCR, synthesis of capsid protein and assembly of infectious progeny virus in the presence of inhibitor blockage. The results indicated that BPV entry into embryonic bovine trachael cells utilizes endocytosis in clathrin-coated vesicles, is dependent upon acidification, and appears to be associated with actin and microtubule dependency. Evidence for viral entry through caveolae was not obtained. These findings provide a fuller understanding of the early cell-entry events of the replication cycle for members of the genus Bocavirus.

  9. Anchored but not internalized: shape dependent endocytosis of nanodiamond

    Science.gov (United States)

    Zhang, Bokai; Feng, Xi; Yin, Hang; Ge, Zhenpeng; Wang, Yanhuan; Chu, Zhiqin; Raabova, Helena; Vavra, Jan; Cigler, Petr; Liu, Renbao; Wang, Yi; Li, Quan

    2017-04-01

    Nanoparticle-cell interactions begin with the cellular uptake of the nanoparticles, a process that eventually determines their cellular fate. In the present work, we show that the morphological features of nanodiamonds (NDs) affect both the anchoring and internalization stages of their endocytosis. While a prickly ND (with sharp edges/corners) has no trouble of anchoring onto the plasma membrane, it suffers from difficult internalization afterwards. In comparison, the internalization of a round ND (obtained by selective etching of the prickly ND) is not limited by its lower anchoring amount and presents a much higher endocytosis amount. Molecular dynamics simulation and continuum modelling results suggest that the observed difference in the anchoring of round and prickly NDs likely results from the reduced contact surface area with the cell membrane of the former, while the energy penalty associated with membrane curvature generation, which is lower for a round ND, may explain its higher probability of the subsequent internalization.

  10. Endocytosis of desmosomal plaques depends on intact actin filaments and leads to a nondegradative compartment

    DEFF Research Database (Denmark)

    Holm, Pernille K.; Hansen, Steen H.; Sandvig, Kirsten

    1993-01-01

    Cellebiologi, human epithelial cell line, growth inhibition, desmosomes, clathrin-independent endocytosis, cytoskeleton, nondegradative compartment......Cellebiologi, human epithelial cell line, growth inhibition, desmosomes, clathrin-independent endocytosis, cytoskeleton, nondegradative compartment...

  11. Voltage Dependence of a Neuromodulator-Activated Ionic Current123

    Science.gov (United States)

    2016-01-01

    Abstract The neuromodulatory inward current (IMI) generated by crab Cancer borealis stomatogastric ganglion neurons is an inward current whose voltage dependence has been shown to be crucial in the activation of oscillatory activity of the pyloric network of this system. It has been previously shown that IMI loses its voltage dependence in conditions of low extracellular calcium, but that this effect appears to be regulated by intracellular calmodulin. Voltage dependence is only rarely regulated by intracellular signaling mechanisms. Here we address the hypothesis that the voltage dependence of IMI is mediated by intracellular signaling pathways activated by extracellular calcium. We demonstrate that calmodulin inhibitors and a ryanodine antagonist can reduce IMI voltage dependence in normal Ca2+, but that, in conditions of low Ca2+, calmodulin activators do not restore IMI voltage dependence. Further, we show evidence that CaMKII alters IMI voltage dependence. These results suggest that calmodulin is necessary but not sufficient for IMI voltage dependence. We therefore hypothesize that the Ca2+/calmodulin requirement for IMI voltage dependence is due to an active sensing of extracellular calcium by a GPCR family calcium-sensing receptor (CaSR) and that the reduction in IMI voltage dependence by a calmodulin inhibitor is due to CaSR endocytosis. Supporting this, preincubation with an endocytosis inhibitor prevented W7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride)-induced loss of IMI voltage dependence, and a CaSR antagonist reduced IMI voltage dependence. Additionally, myosin light chain kinase, which is known to act downstream of the CaSR, seems to play a role in regulating IMI voltage dependence. Finally, a Gβγ-subunit inhibitor also affects IMI voltage dependence, in support of the hypothesis that this process is regulated by a G-protein-coupled CaSR. PMID:27257619

  12. Simian hemorrhagic fever virus cell entry is dependent on CD163 and uses a clathrin-mediated endocytosis-like pathway.

    Science.gov (United States)

    Caì, Yíngyún; Postnikova, Elena N; Bernbaum, John G; Yú, Shu Qìng; Mazur, Steven; Deiuliis, Nicole M; Radoshitzky, Sheli R; Lackemeyer, Matthew G; McCluskey, Adam; Robinson, Phillip J; Haucke, Volker; Wahl-Jensen, Victoria; Bailey, Adam L; Lauck, Michael; Friedrich, Thomas C; O'Connor, David H; Goldberg, Tony L; Jahrling, Peter B; Kuhn, Jens H

    2015-01-01

    Simian hemorrhagic fever virus (SHFV) causes a severe and almost uniformly fatal viral hemorrhagic fever in Asian macaques but is thought to be nonpathogenic for humans. To date, the SHFV life cycle is almost completely uncharacterized on the molecular level. Here, we describe the first steps of the SHFV life cycle. Our experiments indicate that SHFV enters target cells by low-pH-dependent endocytosis. Dynamin inhibitors, chlorpromazine, methyl-β-cyclodextrin, chloroquine, and concanamycin A dramatically reduced SHFV entry efficiency, whereas the macropinocytosis inhibitors EIPA, blebbistatin, and wortmannin and the caveolin-mediated endocytosis inhibitors nystatin and filipin III had no effect. Furthermore, overexpression and knockout study and electron microscopy results indicate that SHFV entry occurs by a dynamin-dependent clathrin-mediated endocytosis-like pathway. Experiments utilizing latrunculin B, cytochalasin B, and cytochalasin D indicate that SHFV does not hijack the actin polymerization pathway. Treatment of target cells with proteases (proteinase K, papain, α-chymotrypsin, and trypsin) abrogated entry, indicating that the SHFV cell surface receptor is a protein. Phospholipases A2 and D had no effect on SHFV entry. Finally, treatment of cells with antibodies targeting CD163, a cell surface molecule identified as an entry factor for the SHFV-related porcine reproductive and respiratory syndrome virus, diminished SHFV replication, identifying CD163 as an important SHFV entry component. Simian hemorrhagic fever virus (SHFV) causes highly lethal disease in Asian macaques resembling human illness caused by Ebola or Lassa virus. However, little is known about SHFV's ecology and molecular biology and the mechanism by which it causes disease. The results of this study shed light on how SHFV enters its target cells. Using electron microscopy and inhibitors for various cellular pathways, we demonstrate that SHFV invades cells by low-pH-dependent, actin

  13. Cellular Response to Titanium Dioxide Nanoparticles in Intestinal Epithelial Caco-2 Cells is Dependent on Endocytosis-Associated Structures and Mediated by EGFR

    Science.gov (United States)

    Krüger, Kristin; Schrader, Katrin; Klempt, Martin

    2017-01-01

    Titanium dioxide (TiO2) is one of the most applied nanomaterials and widely used in food and non-food industries as an additive or coating material (E171). It has been shown that E171 contains up to 37% particles which are smaller than 100 nm and that TiO2 nanoparticles (NPs) induce cytotoxicity and inflammation. Using a nuclear factor Kappa-light-chain enhancer of activated B cells (NF-κB) reporter cell line (Caco-2nfkb-RE), Real time polymerase chain reaction (PCR), and inhibition of dynamin and clathrin, it was shown that cellular responses induced by 5 nm and 10 nm TiO2 NPs (nominal size) depends on endocytic processes. As endocytosis is often dependent on the epithelial growth factor receptor (EGFR), further investigations focused on the involvement of EGFR in the uptake of TiO2 NPs: (1) inhibition of EGFR reduced inflammatory markers of the cell (i.e., nuclear factor (NF)-κB activity, mRNA of IL8, CCL20, and CXCL10); and (2) exposure of Caco-2 cells to TiO2 NPs activated the intracellular EGFR cascade beginning with EGFR-mediated extracellular signal-regulated kinases (ERK)1/2, and including transcription factor ELK1. This was followed by the expression of ERK1/2 target genes CCL2 and CXCL3. We concluded that TiO2 NPs enter the cell via EGFR-associated endocytosis, followed by activation of the EGFR/ERK/ELK signaling pathway, which finally induces NF-κB. No changes in inflammatory response are observed in Caco-2 cells exposed to 32 nm and 490 nm TiO2 particles. PMID:28387727

  14. Cellular Response to Titanium Dioxide Nanoparticles in Intestinal Epithelial Caco-2 Cells is Dependent on Endocytosis-Associated Structures and Mediated by EGFR.

    Science.gov (United States)

    Krüger, Kristin; Schrader, Katrin; Klempt, Martin

    2017-04-07

    Titanium dioxide (TiO₂) is one of the most applied nanomaterials and widely used in food and non-food industries as an additive or coating material (E171). It has been shown that E171 contains up to 37% particles which are smaller than 100 nm and that TiO₂ nanoparticles (NPs) induce cytotoxicity and inflammation. Using a nuclear factor Kappa-light-chain enhancer of activated B cells (NF-κB) reporter cell line (Caco-2 nfkb-RE ), Real time polymerase chain reaction (PCR), and inhibition of dynamin and clathrin, it was shown that cellular responses induced by 5 nm and 10 nm TiO₂ NPs (nominal size) depends on endocytic processes. As endocytosis is often dependent on the epithelial growth factor receptor (EGFR), further investigations focused on the involvement of EGFR in the uptake of TiO₂ NPs: (1) inhibition of EGFR reduced inflammatory markers of the cell (i.e., nuclear factor (NF)-κB activity, mRNA of IL8, CCL20, and CXCL10); and (2) exposure of Caco-2 cells to TiO₂ NPs activated the intracellular EGFR cascade beginning with EGFR-mediated extracellular signal-regulated kinases (ERK)1/2, and including transcription factor ELK1. This was followed by the expression of ERK1/2 target genes CCL2 and CXCL3. We concluded that TiO₂ NPs enter the cell via EGFR-associated endocytosis, followed by activation of the EGFR/ERK/ELK signaling pathway, which finally induces NF-κB. No changes in inflammatory response are observed in Caco-2 cells exposed to 32 nm and 490 nm TiO₂ particles.

  15. The Effect of Substrate Elasticity and Actomyosin Contractility on Different Forms of Endocytosis

    Science.gov (United States)

    Missirlis, Dimitris

    2014-01-01

    Substrate mechanical properties have emerged as potent determinants of cell functions and fate. We here tested the hypothesis that different forms of endocytosis are regulated by the elasticity of the synthetic hydrogels cells are cultured on. Towards this objective, we quantified cell-associated fluorescence of the established endocytosis markers transferrin (Tf) and cholera toxin subunit B (CTb) using a flow-cytometry based protocol, and imaged marker internalization using microscopy techniques. Our results demonstrated that clathrin-mediated endocytosis of Tf following a 10-minute incubation with a fibroblast cell line was lower on the softer substrates studied (5 kPa) compared to those with elasticities of 40 and 85 kPa. This effect was cancelled after 1-hour incubation revealing that intracellular accumulation of Tf at this time point did not depend on substrate elasticity. Lipid-raft mediated endocytosis of CTb, on the other hand, was not affected by substrate elasticity in the studied range of time and substrate elasticity. The use of pharmacologic contractility inhibitors revealed inhibition of endocytosis for both Tf and CTb after a 10-minute incubation and a dissimilar effect after 1 hour depending on the inhibitor type. Further, the internalization of fluorescent NPs, used as model drug delivery systems, showed a dependence on substrate elasticity, while transfection efficiency was unaffected by it. Finally, an independence on substrate elasticity of Tf and CTb association with HeLa cells indicated that there are cell-type differences in this respect. Overall, our results suggest that clathrin-mediated but not lipid-raft mediated endocytosis is potentially influenced by substrate mechanics at the cellular level, while intracellular trafficking and accumulation show a more complex dependence. Our findings are discussed in the context of previous work on how substrate mechanics affect the fundamental process of endocytosis and highlight important

  16. Weak Molecular Interactions in Clathrin-Mediated Endocytosis

    Directory of Open Access Journals (Sweden)

    Sarah M. Smith

    2017-11-01

    Full Text Available Clathrin-mediated endocytosis is a process by which specific molecules are internalized from the cell periphery for delivery to early endosomes. The key stages in this step-wise process, from the starting point of cargo recognition, to the later stage of assembly of the clathrin coat, are dependent on weak interactions between a large network of proteins. This review discusses the structural and functional data that have improved our knowledge and understanding of the main weak molecular interactions implicated in clathrin-mediated endocytosis, with a particular focus on the two key proteins: AP2 and clathrin.

  17. Endocytosis of Corn Oil-Caseinate Emulsions In Vitro: Impacts of Droplet Sizes

    Science.gov (United States)

    Fan, Yuting; Yokoyama, Wally; Yi, Jiang

    2017-01-01

    The relative uptake and mechanisms of lipid-based emulsions of three different particle diameters by Caco-2 cells were studied. The corn oil-sodium caseinate emulsions showed little or no cytotoxicity even at 2 mg/mL protein concentration for any of the three droplet size emulsions. Confocal laser scanning microscopy (CLSM) of Nile red containing emulsions showed that the lipid-based emulsions were absorbed by Caco-2 cells. A negative correlation between the mean droplet size and cellular uptake was observed. There was a time-dependent and energy-dependent uptake as shown by incubation at different times and treatment with sodium azide a general inhibitor of active transport. The endocytosis of lipid-based emulsions was size-dependent. The internalization of nanoemulsion droplets into Caco-2 cells mainly occurred through clathrin- and caveolae/lipid raft-related pathways, while macropinocytosis route played the most important role for 556 nm emulsion endocytosis as shown by the use of specific pathway inhibitors. Permeability of the emulsion through the apical or basal routes also suggested that active transport may be the main route for lipid-based nanoemulsions. The results may assist in the design and application of lipid-based nanoemulsions in nutraceuticals and pharmaceuticals delivery. PMID:29072633

  18. Endocytosis of Corn Oil-Caseinate Emulsions In Vitro: Impacts of Droplet Sizes

    Directory of Open Access Journals (Sweden)

    Yuting Fan

    2017-10-01

    Full Text Available The relative uptake and mechanisms of lipid-based emulsions of three different particle diameters by Caco-2 cells were studied. The corn oil-sodium caseinate emulsions showed little or no cytotoxicity even at 2 mg/mL protein concentration for any of the three droplet size emulsions. Confocal laser scanning microscopy (CLSM of Nile red containing emulsions showed that the lipid-based emulsions were absorbed by Caco-2 cells. A negative correlation between the mean droplet size and cellular uptake was observed. There was a time-dependent and energy-dependent uptake as shown by incubation at different times and treatment with sodium azide a general inhibitor of active transport. The endocytosis of lipid-based emulsions was size-dependent. The internalization of nanoemulsion droplets into Caco-2 cells mainly occurred through clathrin- and caveolae/lipid raft-related pathways, while macropinocytosis route played the most important role for 556 nm emulsion endocytosis as shown by the use of specific pathway inhibitors. Permeability of the emulsion through the apical or basal routes also suggested that active transport may be the main route for lipid-based nanoemulsions. The results may assist in the design and application of lipid-based nanoemulsions in nutraceuticals and pharmaceuticals delivery.

  19. Anti-Inflammatory Action of an Antimicrobial Model Peptide That Suppresses the TRIF-Dependent Signaling Pathway via Inhibition of Toll-Like Receptor 4 Endocytosis in Lipopolysaccharide-Stimulated Macrophages.

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    Do-Wan Shim

    Full Text Available Antimicrobial peptides (AMPs, also called host defense peptides, particularly those with amphipathic helical structures, are emerging as target molecules for therapeutic development due to their immunomodulatory properties. Although the antimicrobial activity of AMPs is known to be exerted primarily by permeation of the bacterial membrane, the mechanism underlying its anti-inflammatory activity remains to be elucidated. We report potent anti-inflammatory activity of WALK11.3, an antimicrobial model peptide with an amphipathic helical conformation, in lipopolysaccharide (LPS-stimulated RAW264.7 cells. This peptide inhibited the expression of inflammatory mediators, including nitric oxide, COX-2, IL-1β, IL-6, INF-β, and TNF-α. Although WALK11.3 did not exert a major effect on all downstream signaling in the MyD88-dependent pathway, toll-like receptor 4 (TLR4- mediated pro-inflammatory signals were markedly attenuated in the TRIF-dependent pathway due to inhibition of the phosphorylation of STAT1 by attenuation of IRF3 phosphorylation. WALK11.3 specifically inhibited the endocytosis of TLR4, which is essential for triggering TRIF-mediated signaling in macrophage cells. Hence, we suggest that specific interference with TLR4 endocytosis could be one of the major modes of the anti-inflammatory action of AMPs. Our designed WALK11 peptides, which possess both antimicrobial and anti-inflammatory activities, may be promising molecules for the development of therapies for infectious inflammation.

  20. Substrate-induced ubiquitylation and endocytosis of yeast amino acid permeases.

    Science.gov (United States)

    Ghaddar, Kassem; Merhi, Ahmad; Saliba, Elie; Krammer, Eva-Maria; Prévost, Martine; André, Bruno

    2014-12-01

    Many plasma membrane transporters are downregulated by ubiquitylation, endocytosis, and delivery to the lysosome in response to various stimuli. We report here that two amino acid transporters of Saccharomyces cerevisiae, the general amino acid permease (Gap1) and the arginine-specific permease (Can1), undergo ubiquitin-dependent downregulation in response to their substrates and that this downregulation is not due to intracellular accumulation of the transported amino acids but to transport catalysis itself. Following an approach based on permease structural modeling, mutagenesis, and kinetic parameter analysis, we obtained evidence that substrate-induced endocytosis requires transition of the permease to a conformational state preceding substrate release into the cell. Furthermore, this transient conformation must be stable enough, and thus sufficiently populated, for the permease to undergo efficient downregulation. Additional observations, including the constitutive downregulation of two active Gap1 mutants altered in cytosolic regions, support the model that the substrate-induced conformational transition inducing endocytosis involves remodeling of cytosolic regions of the permeases, thereby promoting their recognition by arrestin-like adaptors of the Rsp5 ubiquitin ligase. Similar mechanisms might control many other plasma membrane transporters according to the external concentrations of their substrates. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  1. Timely Endocytosis of Cytokinetic Enzymes Prevents Premature Spindle Breakage during Mitotic Exit.

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    Cheen Fei Chin

    2016-07-01

    Full Text Available Cytokinesis requires the spatio-temporal coordination of membrane deposition and primary septum (PS formation at the division site to drive acto-myosin ring (AMR constriction. It has been demonstrated that AMR constriction invariably occurs only after the mitotic spindle disassembly. It has also been established that Chitin Synthase II (Chs2p neck localization precedes mitotic spindle disassembly during mitotic exit. As AMR constriction depends upon PS formation, the question arises as to how chitin deposition is regulated so as to prevent premature AMR constriction and mitotic spindle breakage. In this study, we propose that cells regulate the coordination between spindle disassembly and AMR constriction via timely endocytosis of cytokinetic enzymes, Chs2p, Chs3p, and Fks1p. Inhibition of endocytosis leads to over accumulation of cytokinetic enzymes during mitotic exit, which accelerates the constriction of the AMR, and causes spindle breakage that eventually could contribute to monopolar spindle formation in the subsequent round of cell division. Intriguingly, the mitotic spindle breakage observed in endocytosis mutants can be rescued either by deleting or inhibiting the activities of, CHS2, CHS3 and FKS1, which are involved in septum formation. The findings from our study highlight the importance of timely endocytosis of cytokinetic enzymes at the division site in safeguarding mitotic spindle integrity during mitotic exit.

  2. ER network homeostasis is critical for plant endosome streaming and endocytosis

    Science.gov (United States)

    Stefano, Giovanni; Renna, Luciana; Lai, YaShiuan; Slabaugh, Erin; Mannino, Nicole; Buono, Rafael A; Otegui, Marisa S; Brandizzi, Federica

    2015-01-01

    Eukaryotic cells internalize cargo at the plasma membrane via endocytosis, a vital process that is accomplished through a complex network of endosomal organelles. In mammalian cells, the ER is in close association with endosomes and regulates their fission. Nonetheless, the physiological role of such interaction on endocytosis is yet unexplored. Here, we probed the existence of ER–endosome association in plant cells and assayed its physiological role in endocytosis. Through live-cell imaging and electron microscopy studies, we established that endosomes are extensively associated with the plant ER, supporting conservation of interaction between heterotypic organelles in evolutionarily distant kingdoms. Furthermore, by analyzing ER–endosome dynamics in genetic backgrounds with defects in ER structure and movement, we also established that the ER network integrity is necessary for homeostasis of the distribution and streaming of various endosome populations as well as for efficient endocytosis. These results support a novel model that endocytosis homeostasis depends on a spatiotemporal control of the endosome dynamics dictated by the ER membrane network. PMID:27462431

  3. Alpha-synuclein induces lysosomal rupture and cathepsin dependent reactive oxygen species following endocytosis.

    Directory of Open Access Journals (Sweden)

    David Freeman

    Full Text Available α-synuclein dysregulation is a critical aspect of Parkinson's disease pathology. Recent studies have observed that α-synuclein aggregates are cytotoxic to cells in culture and that this toxicity can be spread between cells. However, the molecular mechanisms governing this cytotoxicity and spread are poorly characterized. Recent studies of viruses and bacteria, which achieve their cytoplasmic entry by rupturing intracellular vesicles, have utilized the redistribution of galectin proteins as a tool to measure vesicle rupture by these organisms. Using this approach, we demonstrate that α-synuclein aggregates can induce the rupture of lysosomes following their endocytosis in neuronal cell lines. This rupture can be induced by the addition of α-synuclein aggregates directly into cells as well as by cell-to-cell transfer of α-synuclein. We also observe that lysosomal rupture by α-synuclein induces a cathepsin B dependent increase in reactive oxygen species (ROS in target cells. Finally, we observe that α-synuclein aggregates can induce inflammasome activation in THP-1 cells. Lysosomal rupture is known to induce mitochondrial dysfunction and inflammation, both of which are well established aspects of Parkinson's disease, thus connecting these aspects of Parkinson's disease to the propagation of α-synuclein pathology in cells.

  4. Recording the dynamic endocytosis of single gold nanoparticles by AFM-based force tracing.

    Science.gov (United States)

    Ding, Bohua; Tian, Yongmei; Pan, Yangang; Shan, Yuping; Cai, Mingjun; Xu, Haijiao; Sun, Yingchun; Wang, Hongda

    2015-05-07

    We utilized force tracing to directly record the endocytosis of single gold nanoparticles (Au NPs) with different sizes, revealing the size-dependent endocytosis dynamics and the crucial role of membrane cholesterol. The force, duration and velocity of Au NP invagination are accurately determined at the single-particle and microsecond level unprecedentedly.

  5. Characterization of endocytosis and exocytosis of cationic nanoparticles in airway epithelium cells

    Energy Technology Data Exchange (ETDEWEB)

    Dombu, Christophe Youta; Kroubi, Maya; Zibouche, Rima; Matran, Regis; Betbeder, Didier, E-mail: dbetbeder@aol.com [EA 4483, IFR 114, Laboratoire de Physiologie, Faculte de Medecine Pole Recherche, Universite de Lille 2, 1 place de Verdun, 59045 Lille Cedex (France)

    2010-09-03

    A major challenge of drug delivery using colloids via the airway is to understand the mechanism implied in their interactions with epithelial cells. The purpose of this work was to characterize the process of endocytosis and exocytosis of cationic nanoparticles (NPs) made of maltodextrin which were developed as a delivery system for antigens in vaccine applications. Confocal microscopy demonstrated that these NP are rapidly endocytosed after as little as 3 min incubation, and that the endocytosis was also faster than NP binding since most of the NPs were found in the middle of the cells around the nuclei. A saturation limit was observed after a 40 min incubation, probably due to an equilibrium becoming established between endocytosis and exocytosis. Endocytosis was dramatically reduced at 4 deg. C compared with 37 deg. C, or by NaN{sub 3} treatment, both results suggesting an energy dependent process. Protamine pretreatment of the cells inhibited NPs uptake and we found that clathrin pathway is implied in their endocytosis. Cholesterol depletion increased NP uptake by 300% and this phenomenon was explained by the fact that cholesterol depletion totally blocked NP exocytosis. These results suggest that these cationic NPs interact with anionic sites, are quickly endocytosed via the clathrin pathway and that their exocytosis is cholesterol dependent, and are similar to those obtained in other studies with viruses such as influenza.

  6. Simian Hemorrhagic Fever Virus Cell Entry Is Dependent on CD163 and Uses a Clathrin-Mediated Endocytosis-Like Pathway

    OpenAIRE

    Caì, Yíngyún; Postnikova, Elena N.; Bernbaum, John G.; Yú, Shuǐqìng; Mazur, Steven; Deiuliis, Nicole M.; Radoshitzky, Sheli R.; Lackemeyer, Matthew G.; McCluskey, Adam; Robinson, Phillip J.; Haucke, Volker; Wahl-Jensen, Victoria; Bailey, Adam L.; Lauck, Michael; Friedrich, Thomas C.

    2014-01-01

    Simian hemorrhagic fever virus (SHFV) causes a severe and almost uniformly fatal viral hemorrhagic fever in Asian macaques but is thought to be nonpathogenic for humans. To date, the SHFV life cycle is almost completely uncharacterized on the molecular level. Here, we describe the first steps of the SHFV life cycle. Our experiments indicate that SHFV enters target cells by low-pH-dependent endocytosis. Dynamin inhibitors, chlorpromazine, methyl-β-cyclodextrin, chloroquine, and concanamycin A ...

  7. State-dependent bulk-boundary maps and black hole complementarity

    NARCIS (Netherlands)

    Papadodimas, Kyriakos; Raju, Suvrat

    2014-01-01

    We provide a simple and explicit construction of local bulk operators that describe the interior of a black hole in the AdS/CFT correspondence. The existence of these operators is predicated on the assumption that the mapping of CFT operators to local bulk operators depends on the state of the CFT.

  8. Herpes simplex virus internalization into epithelial cells requires Na+/H+ exchangers and p21-activated kinases but neither clathrin- nor caveolin-mediated endocytosis.

    Science.gov (United States)

    Devadas, Deepika; Koithan, Thalea; Diestel, Randi; Prank, Ute; Sodeik, Beate; Döhner, Katinka

    2014-11-01

    Herpes simplex virus 1 (HSV-1) is an alphaherpesvirus that has been reported to infect some epithelial cell types by fusion at the plasma membrane but others by endocytosis. To determine the molecular mechanisms of productive HSV-1 cell entry, we perturbed key endocytosis host factors using specific inhibitors, RNA interference (RNAi), or overexpression of dominant negative proteins and investigated their effects on HSV-1 infection in the permissive epithelial cell lines Vero, HeLa, HEp-2, and PtK2. HSV-1 internalization required neither endosomal acidification nor clathrin- or caveolin-mediated endocytosis. In contrast, HSV-1 gene expression and internalization were significantly reduced after treatment with 5-(N-ethyl-N-isopropyl)amiloride (EIPA). EIPA blocks the activity of Na(+)/H(+) exchangers, which are plasma membrane proteins implicated in all forms of macropinocytosis. HSV-1 internalization furthermore required the function of p21-activated kinases that contribute to macropinosome formation. However, in contrast to some forms of macropinocytosis, HSV-1 did not enlist the activities of protein kinase C (PKC), tyrosine kinases, C-terminal binding protein 1, or dynamin to activate its internalization. These data suggest that HSV-1 depends on Na(+)/H(+) exchangers and p21-activated kinases either for macropinocytosis or for local actin rearrangements required for fusion at the plasma membrane or subsequent passage through the actin cortex underneath the plasma membrane. After initial replication in epithelial cells, herpes simplex viruses (HSVs) establish latent infections in neurons innervating these regions. Upon primary infection and reactivation from latency, HSVs cause many human skin and neurological diseases, particularly in immunocompromised hosts, despite the availability of effective antiviral drugs. Many viruses use macropinocytosis for virus internalization, and many host factors mediating this entry route have been identified, although the

  9. Gap-related trapped magnetic flux dependence between single and combined bulk superconductors

    International Nuclear Information System (INIS)

    Deng, Z.; Miki, M.; Felder, B.; Tsuzuki, K.; Shinohara, N.; Uetake, T.; Izumi, M.

    2011-01-01

    Highlights: → Rectangular YBCO bulks to realize a compact combination. → The gap effect was added to consider in the trapped flux density mapping. → The trapped-flux dependence between single and combined bulks is gap related. → It is possible to estimate the total magnetic flux of bulk combinations. - Abstract: Aiming at examining the trapped-flux dependence between single and combined bulk superconductors for field-pole applications, three rectangular Y 1.65 Ba 2 Cu 3 O 7-x (YBCO) bulks with a possibly compact combination were employed to investigate the trapped-flux characteristics of single and combined bulks with a field-cooling magnetization (FCM) method. A gap-related dependence was found between them. At lower gaps of 1 mm and 5 mm, the peak trapped fields and total magnetic flux of combined bulks are both smaller than the additive values of each single bulk, which can be ascribed to the demagnetization influences of the field around the bulk generated by the adjacent ones. While, at larger gaps like 10 mm, the situation becomes reversed. The combined bulks can attain bigger peak trapped fields as well as total magnetic flux, which indicates that the magnetic field by the bulk combination can reach higher gaps, thanks to the bigger magnetic energy compared with the single bulk. The presented results show that, on one hand, it is possible to estimate the total trapped magnetic flux of combined bulks by an approximate additive method of each single bulk while considering a demagnetization factor; on the other hand, it also means that the performance of combined bulks will be superior to the addition of each single bulk at larger gaps, thus preferable for large-scaled magnet applications.

  10. Gap-related trapped magnetic flux dependence between single and combined bulk superconductors

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Z., E-mail: zgdeng@gmail.co [Laboratory of Applied Physics, Department of Marine Electronics and Mechanical Engineering, Tokyo University of Marine Science and Technology, Tokyo 135-8533 (Japan); Miki, M.; Felder, B.; Tsuzuki, K.; Shinohara, N.; Uetake, T.; Izumi, M. [Laboratory of Applied Physics, Department of Marine Electronics and Mechanical Engineering, Tokyo University of Marine Science and Technology, Tokyo 135-8533 (Japan)

    2011-05-15

    Highlights: {yields} Rectangular YBCO bulks to realize a compact combination. {yields} The gap effect was added to consider in the trapped flux density mapping. {yields} The trapped-flux dependence between single and combined bulks is gap related. {yields} It is possible to estimate the total magnetic flux of bulk combinations. - Abstract: Aiming at examining the trapped-flux dependence between single and combined bulk superconductors for field-pole applications, three rectangular Y{sub 1.65}Ba{sub 2}Cu{sub 3}O{sub 7-x} (YBCO) bulks with a possibly compact combination were employed to investigate the trapped-flux characteristics of single and combined bulks with a field-cooling magnetization (FCM) method. A gap-related dependence was found between them. At lower gaps of 1 mm and 5 mm, the peak trapped fields and total magnetic flux of combined bulks are both smaller than the additive values of each single bulk, which can be ascribed to the demagnetization influences of the field around the bulk generated by the adjacent ones. While, at larger gaps like 10 mm, the situation becomes reversed. The combined bulks can attain bigger peak trapped fields as well as total magnetic flux, which indicates that the magnetic field by the bulk combination can reach higher gaps, thanks to the bigger magnetic energy compared with the single bulk. The presented results show that, on one hand, it is possible to estimate the total trapped magnetic flux of combined bulks by an approximate additive method of each single bulk while considering a demagnetization factor; on the other hand, it also means that the performance of combined bulks will be superior to the addition of each single bulk at larger gaps, thus preferable for large-scaled magnet applications.

  11. Excessive Cellular S-nitrosothiol Impairs Endocytosis of Auxin Efflux Transporter PIN2

    Directory of Open Access Journals (Sweden)

    Min Ni

    2017-11-01

    Full Text Available S-nitrosoglutathione reductase (GSNOR1 is the key enzyme that regulates cellular levels of S-nitrosylation across kingdoms. We have previously reported that loss of GSNOR1 resulted in impaired auxin signaling and compromised auxin transport in Arabidopsis, leading to the auxin-related morphological phenotypes. However, the molecular mechanism underpinning the compromised auxin transport in gsnor1-3 mutant is still unknown. Endocytosis of plasma-membrane (PM-localized efflux PIN proteins play critical roles in auxin transport. Therefore, we investigate whether loss of GSNOR1 function has any effects on the endocytosis of PIN-FORMED (PIN proteins. It was found that the endocytosis of either the endogenous PIN2 or the transgenically expressed PIN2-GFP was compromised in the root cells of gsnor1-3 seedlings relative to Col-0. The internalization of PM-associated PIN2 or PIN2-GFP into Brefeldin A (BFA bodies was significantly reduced in gsnor1-3 upon BFA treatment in a manner independent of de novo protein synthesis. In addition, the exogenously applied GSNO not only compromised the endocytosis of PIN2-GFP but also inhibited the root elongation in a concentration-dependent manner. Taken together, our results indicate that, besides the reduced PIN2 level, one or more compromised components in the endocytosis pathway could account for the reduced endocytosis of PIN2 in gsnor1-3.

  12. Endocytosis and Endosomal Trafficking in Plants.

    Science.gov (United States)

    Paez Valencia, Julio; Goodman, Kaija; Otegui, Marisa S

    2016-04-29

    Endocytosis and endosomal trafficking are essential processes in cells that control the dynamics and turnover of plasma membrane proteins, such as receptors, transporters, and cell wall biosynthetic enzymes. Plasma membrane proteins (cargo) are internalized by endocytosis through clathrin-dependent or clathrin-independent mechanism and delivered to early endosomes. From the endosomes, cargo proteins are recycled back to the plasma membrane via different pathways, which rely on small GTPases and the retromer complex. Proteins that are targeted for degradation through ubiquitination are sorted into endosomal vesicles by the ESCRT (endosomal sorting complex required for transport) machinery for degradation in the vacuole. Endocytic and endosomal trafficking regulates many cellular, developmental, and physiological processes, including cellular polarization, hormone transport, metal ion homeostasis, cytokinesis, pathogen responses, and development. In this review, we discuss the mechanisms that mediate the recognition and sorting of endocytic and endosomal cargos, the vesiculation processes that mediate their trafficking, and their connection to cellular and physiological responses in plants.

  13. Interaction among Saccharomyces cerevisiae pheromone receptors during endocytosis

    Directory of Open Access Journals (Sweden)

    Chien-I Chang

    2014-03-01

    Full Text Available This study investigates endocytosis of Saccharomyces cerevisiae α-factor receptor and the role that receptor oligomerization plays in this process. α-factor receptor contains signal sequences in the cytoplasmic C-terminal domain that are essential for ligand-mediated endocytosis. In an endocytosis complementation assay, we found that oligomeric complexes of the receptor undergo ligand-mediated endocytosis when the α-factor binding site and the endocytosis signal sequences are located in different receptors. Both in vitro and in vivo assays suggested that ligand-induced conformational changes in one Ste2 subunit do not affect neighboring subunits. Therefore, recognition of the endocytosis signal sequence and recognition of the ligand-induced conformational change are likely to be two independent events.

  14. Chirality-dependent friction of bulk molecular solids.

    Science.gov (United States)

    Yang, Dian; Cohen, Adam E

    2014-08-26

    We show that the solid-solid friction between bulk chiral molecular solids can depend on the relative chirality of the two materials. In menthol and 1-phenyl-1-butanol, heterochiral friction is smaller than homochiral friction, while in ibuprofen, heterochiral friction is larger. Chiral asymmetries in the coefficient of sliding friction vary with temperature and can be as large as 30%. In the three compounds tested, the sign of the difference between heterochiral and homochiral friction correlated with the sign of the difference in melting point between racemate (compound or conglomerate) and pure enantiomer. Menthol and ibuprofen each form a stable racemic compound, while 1-phenyl-1-butanol forms a racemic conglomerate. Thus, a difference between heterochiral and homochiral friction does not require the formation of a stable interfacial racemic compound. Measurements of chirality-dependent friction provide a unique means to distinguish the role of short-range intermolecular forces from all other sources of dissipation in the friction of bulk molecular solids.

  15. Endocytosis regulates membrane localization and function of the fusogen EFF-1.

    Science.gov (United States)

    Smurova, Ksenia; Podbilewicz, Benjamin

    2017-07-03

    Cell fusion is essential for sexual reproduction and formation of muscles, bones, and placenta. Two families of cell fusion proteins (Syncytins and FFs) have been identified in eukaryotes. Syncytins have been shown to form the giant syncytial trophoblasts in the placenta. The FFs are essential to fuse cells in the skin, reproductive, excretory, digestive and nervous systems in nematodes. EFF-1 (Epithelial Fusion Failure 1), a member of the FF family, is a type I membrane glycoprotein that is essential for most cell fusions in C. elegans. The crystal structure of EFF-1 ectodomain reveals striking structural similarity to class II fusion glycoproteins from enveloped viruses (e.g. dengue and rubella) that mediate virus to cell fusion. We found EFF-1 to be present on the plasma membrane and in RAB-5-positive early endosomes, with EFF-1 recycling between these 2 cell compartments. Only when EFF-1 proteins transiently arrive to the surfaces of 2 adjacent cells do they dynamically interact in trans and mediate membrane fusion. EFF-1 is continuously internalized by receptor-mediated endocytosis via the activity of 2 small GTPases: RAB-5 and Dynamin. Here we propose a model that explains how EFF-1 endocytosis together with interactions in trans can control cell-cell fusion. Kontani et al. showed that vacuolar ATPase (vATPase) mutations result in EFF-1-dependent hyperfusion. 1 We propose that vATPase is required for normal degradation of EFF-1. Failure to degrade EFF-1 results in delayed hyperfusion and mislocalization to organelles that appear to be recycling endosomes. EFF-1 is also required to fuse neurons as part of the repair mechanism following injury and to prune dendrites. We speculate that EFF-1 may regulate neuronal tree like structures via endocytosis. Thus, endocytosis of cell-cell fusion proteins functions to prevent merging of cells and to sculpt organs and neurons.

  16. Size-dependent internalisation of folate-decorated nanoparticles via the pathways of clathrin and caveolae-mediated endocytosis in ARPE-19 cells.

    Science.gov (United States)

    Langston Suen, Wai-Leung; Chau, Ying

    2014-04-01

    We aim to quantify the effect of size and degree of folate loading of folate-decorated polymeric nanoparticles (NPs) on the kinetics of cellular uptake and the selection of endocytic pathways in retinal pigment epithelium (RPE) cells. In this study, methoxy-poly(ethylene glycol)-b-polycaprolactone (mPEG-b-PCL) and folate-functionalized PEG-b-PCL were synthesized for assembling into nanoparticles with sizes ranging from 50 nm to 250 nm. These nanoparticles were internalized into ARPE-19 (human RPE cell line) via receptor-mediated endocytosis. A two-step endocytosis process mathematical model was adopted to quantify binding affinity and uptake kinetics of nanoparticles in RPE cells in uptake and inhibition studies. Nanoparticles with 100% folate loading have highest binding affinity and uptake rate in RPE cells. Maximum uptake rate (Vmax) of nanoparticles increased as the size of particles decreased from 250 nm to 50 nm. Endocytic pathway study was studied by using chlorpromazine and methyl-β-cyclodextran (MβCD), which are clathrin- and caveolae-mediated endocytosis inhibitors, respectively. Both chlorpromazine and MβCD inhibited the uptake of folate-decorated nanoparticles. Inhibition constant (Ki) and maximum uptake rate (Vmax) revealed that 50 nm and 120 nm folate-decorated nanoparticles were found to be internalized via both clathrin- and caveolae-mediated endocytosis. The 250 nm folate-decorated nanoparticles, however, were only internalized via caveolae-mediated pathway. Increased uptake rate of folate-decorated NPs into RPE cells is observed with increasing degree of folate modification. These NPs utilize both clathrin- and caveolae-mediated receptor-mediated endocytosis pathways to enter RPE cells upon size variation. The 50 nm NPs are internalized the fastest, with clathrin-mediated endocytosis as the preferred route. Uptake of 250 nm particles is the slowest and is dominated by caveolae-mediated endocytosis. © 2013 Royal Pharmaceutical

  17. Bladder uptake of liposomes after intravesical administration occurs by endocytosis.

    Directory of Open Access Journals (Sweden)

    Bharathi Raja Rajaganapathy

    Full Text Available Liposomes have been used therapeutically and as a local drug delivery system in the bladder. However, the exact mechanism for the uptake of liposomes by bladder cells is unclear. In the present study, we investigated the role of endocytosis in the uptake of liposomes by cultured human UROtsa cells of urothelium and rat bladder. UROtsa cells were incubated in serum-free media with liposomes containing colloidal gold particles for 2 h either at 37°C or at 4°C. Transmission Electron Microscopy (TEM images of cells incubated at 37°C found endocytic vesicles containing gold inside the cells. In contrast, only extracellular binding was noticed in cells incubated with liposomes at 4°C. Absence of liposome internalization at 4°C indicates the need of energy dependent endocytosis as the primary mechanism of entry of liposomes into the urothelium. Flow cytometry analysis revealed that the uptake of liposomes at 37°C occurs via clathrin mediated endocytosis. Based on these observations, we propose that clathrin mediated endocytosis is the main route of entry for liposomes into the urothelial layer of the bladder and the findings here support the usefulness of liposomes in intravesical drug delivery.

  18. Gap-related trapped magnetic flux dependence between single and combined bulk superconductors

    Science.gov (United States)

    Deng, Z.; Miki, M.; Felder, B.; Tsuzuki, K.; Shinohara, N.; Uetake, T.; Izumi, M.

    2011-05-01

    Aiming at examining the trapped-flux dependence between single and combined bulk superconductors for field-pole applications, three rectangular Y 1.65Ba 2Cu 3O 7-x (YBCO) bulks with a possibly compact combination were employed to investigate the trapped-flux characteristics of single and combined bulks with a field-cooling magnetization (FCM) method. A gap-related dependence was found between them. At lower gaps of 1 mm and 5 mm, the peak trapped fields and total magnetic flux of combined bulks are both smaller than the additive values of each single bulk, which can be ascribed to the demagnetization influences of the field around the bulk generated by the adjacent ones. While, at larger gaps like 10 mm, the situation becomes reversed. The combined bulks can attain bigger peak trapped fields as well as total magnetic flux, which indicates that the magnetic field by the bulk combination can reach higher gaps, thanks to the bigger magnetic energy compared with the single bulk. The presented results show that, on one hand, it is possible to estimate the total trapped magnetic flux of combined bulks by an approximate additive method of each single bulk while considering a demagnetization factor; on the other hand, it also means that the performance of combined bulks will be superior to the addition of each single bulk at larger gaps, thus preferable for large-scaled magnet applications.

  19. Simulation of Field Dependence of Critical Current Densities of Bulk High Tc Superconducting Materials regarding Thermally Activated Flux Motion

    Science.gov (United States)

    Santosh, M.; Naik, S. Pavan Kumar; Koblischka, M. R.

    2017-07-01

    In the upcoming generation, bulk high temperature superconductors (HTS) will play a crucial and a promising role in numerous industrial applications ranging from Maglev trains to magnetic resonance imaging, etc. Especially, the bulk HTS as permanent magnets are suitable due to the fact that they can trap magnetic fields being several orders of magnitude higher than those of the best hard ferromagnets. The bulk HTS LREBa2Cu3O7-δ (LREBCO or LRE-123, LRE: Y, Gd, etc.,) materials could obtain very powerful compact superconducting super-magnets, which can be operated at the cheaper liquid nitrogen temperature or below due to higher critical temperatures (i.e., ∼90 K). As a result, the new advanced technology can be utilized in a more attractive manner for a variety of technological and medical applications which have the capacity to revolutionize the field. An understanding of the magnetic field dependence of the critical current density (J c(H)) is important to develop better adapted materials. To achieve this goal, a variety of Jc (H) behaviours of bulk LREBCO samples were modelled regarding thermally activated flux motion. In essence, the Jc (H) curves follows a certain criterion where an exponential model is applied. However, to fit the complete Jc (H) curve of the LRE-123 samples an unique model is necessary to explain the behavior at low and high fields. The modelling of the various superconducting materials could be understood in terms of the pinning mechanisms.

  20. Endocytosis of HERG is clathrin-independent and involves arf6.

    Directory of Open Access Journals (Sweden)

    Rucha Karnik

    Full Text Available The hERG potassium channel is critical for repolarisation of the cardiac action potential. Reduced expression of hERG at the plasma membrane, whether caused by hereditary mutations or drugs, results in long QT syndrome and increases the risk of ventricular arrhythmias. Thus, it is of fundamental importance to understand how the density of this channel at the plasma membrane is regulated. We used antibodies to an extracellular native or engineered epitope, in conjunction with immunofluorescence and ELISA, to investigate the mechanism of hERG endocytosis in recombinant cells and validated the findings in rat neonatal cardiac myocytes. The data reveal that this channel undergoes rapid internalisation, which is inhibited by neither dynasore, an inhibitor of dynamin, nor a dominant negative construct of Rab5a, into endosomes that are largely devoid of the transferrin receptor. These results support a clathrin-independent mechanism of endocytosis and exclude involvement of dynamin-dependent caveolin and RhoA mechanisms. In agreement, internalised hERG displayed marked overlap with glycosylphosphatidylinositol-anchored GFP, a clathrin-independent cargo. Endocytosis was significantly affected by cholesterol extraction with methyl-β-cyclodextrin and inhibition of Arf6 function with dominant negative Arf6-T27N-eGFP. Taken together, we conclude that hERG undergoes clathrin-independent endocytosis via a mechanism involving Arf6.

  1. Endocytosis of G protein-coupled receptors is regulated by clathrin light chain phosphorylation.

    Science.gov (United States)

    Ferreira, Filipe; Foley, Matthew; Cooke, Alex; Cunningham, Margaret; Smith, Gemma; Woolley, Robert; Henderson, Graeme; Kelly, Eamonn; Mundell, Stuart; Smythe, Elizabeth

    2012-08-07

    Signaling by transmembrane receptors such as G protein-coupled receptors (GPCRs) occurs at the cell surface and throughout the endocytic pathway, and signaling from the cell surface may differ in magnitude and downstream output from intracellular signaling. As a result, the rate at which signaling molecules traverse the endocytic pathway makes a significant contribution to downstream output. Modulation of the core endocytic machinery facilitates differential uptake of individual cargoes. Clathrin-coated pits are a major entry portal where assembled clathrin forms a lattice around invaginating buds that have captured endocytic cargo. Clathrin assembles into triskelia composed of three clathrin heavy chains and associated clathrin light chains (CLCs). Despite the identification of clathrin-coated pits at the cell surface over 30 years ago, the functions of CLCs in endocytosis have been elusive. In this work, we identify a novel role for CLCs in the regulated endocytosis of specific cargoes. Small interfering RNA-mediated knockdown of either CLCa or CLCb inhibits the uptake of GPCRs. Moreover, we demonstrate that phosphorylation of Ser204 in CLCb is required for efficient endocytosis of a subset of GPCRs and identify G protein-coupled receptor kinase 2 (GRK2) as a kinase that can phosphorylate CLCb on Ser204. Overexpression of CLCb(S204A) specifically inhibits the endocytosis of those GPCRs whose endocytosis is GRK2-dependent. Together, these results indicate that CLCb phosphorylation acts as a discriminator for the endocytosis of specific GPCRs. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Endocytosis of collagen by hepatic stellate cells regulates extracellular matrix dynamics.

    Science.gov (United States)

    Bi, Yan; Mukhopadhyay, Dhriti; Drinane, Mary; Ji, Baoan; Li, Xing; Cao, Sheng; Shah, Vijay H

    2014-10-01

    Hepatic stellate cells (HSCs) generate matrix, which in turn may also regulate HSCs function during liver fibrosis. We hypothesized that HSCs may endocytose matrix proteins to sense and respond to changes in microenvironment. Primary human HSCs, LX2, or mouse embryonic fibroblasts (MEFs) [wild-type; c-abl(-/-); or Yes, Src, and Fyn knockout mice (YSF(-/-))] were incubated with fluorescent-labeled collagen or gelatin. Fluorescence-activated cell sorting analysis and confocal microscopy were used for measuring cellular internalization of matrix proteins. Targeted PCR array and quantitative real-time PCR were used to evaluate gene expression changes. HSCs and LX2 cells endocytose collagens in a concentration- and time-dependent manner. Endocytosed collagen colocalized with Dextran 10K, a marker of macropinocytosis, and 5-ethylisopropyl amiloride, an inhibitor of macropinocytosis, reduced collagen internalization by 46%. Cytochalasin D and ML7 blocked collagen internalization by 47% and 45%, respectively, indicating that actin and myosin are critical for collagen endocytosis. Wortmannin and AKT inhibitor blocked collagen internalization by 70% and 89%, respectively, indicating that matrix macropinocytosis requires phosphoinositide-3-kinase (PI3K)/AKT signaling. Overexpression of dominant-negative dynamin-2 K44A blocked matrix internalization by 77%, indicating a role for dynamin-2 in matrix macropinocytosis. Whereas c-abl(-/-) MEF showed impaired matrix endocytosis, YSF(-/-) MEF surprisingly showed increased matrix endocytosis. It was also associated with complex gene regulations that related with matrix dynamics, including increased matrix metalloproteinase 9 (MMP-9) mRNA levels and zymographic activity. HSCs endocytose matrix proteins through macropinocytosis that requires a signaling network composed of PI3K/AKT, dynamin-2, and c-abl. Interaction with extracellular matrix regulates matrix dynamics through modulating multiple gene expressions including MMP-9

  3. Temperature dependence of bulk viscosity in water using acoustic spectroscopy

    International Nuclear Information System (INIS)

    Holmes, M J; Parker, N G; Povey, M J W

    2011-01-01

    Despite its fundamental role in the dynamics of compressible fluids, bulk viscosity has received little experimental attention and there remains a paucity of measured data. Acoustic spectroscopy provides a robust and accurate approach to measuring this parameter. Working from the Navier-Stokes model of a compressible fluid one can show that the bulk viscosity makes a significant and measurable contribution to the frequency-squared acoustic attenuation. Here we employ this methodology to determine the bulk viscosity of Millipore water over a temperature range of 7 to 50 0 C. The measured attenuation spectra are consistent with the theoretical predictions, while the bulk viscosity of water is found to be approximately three times larger than its shear counterpart, reinforcing its significance in acoustic propagation. Moreover, our results demonstrate that this technique can be readily and generally applied to fluids to accurately determine their temperature dependent bulk viscosities.

  4. Sample dependent response of a LaCl{sub 3}:Ce detector in prompt gamma neutron activation analysis of bulk hydrocarbon samples

    Energy Technology Data Exchange (ETDEWEB)

    Naqvi, A.A., E-mail: aanaqvi@kfupm.edu.sa [Department of Physics, King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Al-Matouq, Faris A.; Khiari, F.Z. [Department of Physics, King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Isab, A.A. [Department of Chemistry, King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Khateeb-ur-Rehman,; Raashid, M. [Department of Physics, King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia)

    2013-08-11

    The response of a LaCl{sub 3}:Ce detector has been found to depend upon the hydrogen content of bulk samples in prompt gamma analysis using 14 MeV neutron inelastic scattering. The moderation of 14 MeV neutrons from hydrogen in the bulk sample produces thermal neutrons around the sample which ultimately excite chlorine capture gamma rays in the LaCl{sub 3}:Ce detector material. Interference of 6.11 MeV chlorine gamma rays from the detector itself with 6.13 MeV oxygen gamma rays from the bulk samples makes the intensity of the 6.13 MeV oxygen gamma ray peak relatively insensitive to variations in oxygen concentration. The strong dependence of the 1.95 MeV doublet chlorine gamma ray yield on hydrogen content of the bulk samples confirms fast neutron moderation from hydrogen in the bulk samples as a major source of production of thermal neutrons and chlorine gamma rays in the LaCl{sub 3}:Ce detector material. Despite their poor oxygen detection capabilities, these detectors have nonetheless excellent detection capabilities for hydrogen and carbon in benzene, butyl alcohol, propanol, propanic acid, and formic acid bulk samples using 14 MeV neutron inelastic scattering.

  5. The temperature dependence of the isothermal bulk modulus at 1 bar pressure

    International Nuclear Information System (INIS)

    Garai, J.; Laugier, A.

    2007-01-01

    It is well established that the product of the volume coefficient of thermal expansion and the bulk modulus is nearly constant at temperatures higher than the Debye temperature. Using this approximation allows predicting the values of the bulk modulus. The derived analytical solution for the temperature dependence of the isothermal bulk modulus has been applied to ten substances. The good correlations to the experiments indicate that the expression may be useful for substances for which bulk modulus data are lacking

  6. Endocytosis of hERG Is Clathrin-Independent and Involves Arf6

    Science.gov (United States)

    Abuarab, Nada; Smith, Andrew J.; Hardy, Matthew E. L.; Elliott, David J. S.; Sivaprasadarao, Asipu

    2013-01-01

    The hERG potassium channel is critical for repolarisation of the cardiac action potential. Reduced expression of hERG at the plasma membrane, whether caused by hereditary mutations or drugs, results in long QT syndrome and increases the risk of ventricular arrhythmias. Thus, it is of fundamental importance to understand how the density of this channel at the plasma membrane is regulated. We used antibodies to an extracellular native or engineered epitope, in conjunction with immunofluorescence and ELISA, to investigate the mechanism of hERG endocytosis in recombinant cells and validated the findings in rat neonatal cardiac myocytes. The data reveal that this channel undergoes rapid internalisation, which is inhibited by neither dynasore, an inhibitor of dynamin, nor a dominant negative construct of Rab5a, into endosomes that are largely devoid of the transferrin receptor. These results support a clathrin-independent mechanism of endocytosis and exclude involvement of dynamin-dependent caveolin and RhoA mechanisms. In agreement, internalised hERG displayed marked overlap with glycosylphosphatidylinositol-anchored GFP, a clathrin-independent cargo. Endocytosis was significantly affected by cholesterol extraction with methyl-β-cyclodextrin and inhibition of Arf6 function with dominant negative Arf6-T27N-eGFP. Taken together, we conclude that hERG undergoes clathrin-independent endocytosis via a mechanism involving Arf6. PMID:24392021

  7. Endocytosis of Cytotoxic Granules Is Essential for Multiple Killing of Target Cells by T Lymphocytes.

    Science.gov (United States)

    Chang, Hsin-Fang; Bzeih, Hawraa; Schirra, Claudia; Chitirala, Praneeth; Halimani, Mahantappa; Cordat, Emmanuelle; Krause, Elmar; Rettig, Jens; Pattu, Varsha

    2016-09-15

    CTLs are serial killers that kill multiple target cells via exocytosis of cytotoxic granules (CGs). CG exocytosis is tightly regulated and has been investigated in great detail; however, whether CG proteins are endocytosed following exocytosis and contribute to serial killing remains unknown. By using primary CTLs derived from a knock-in mouse of the CG membrane protein Synaptobrevin2, we show that CGs are endocytosed in a clathrin- and dynamin-dependent manner. Following acidification, endocytosed CGs are recycled through early and late, but not recycling endosomes. CGs are refilled with granzyme B at the late endosome stage and polarize to subsequent synapses formed between the CTL and new target cells. Importantly, inhibiting CG endocytosis in CTLs results in a significant reduction of their cytotoxic activity. Thus, our data demonstrate that continuous endocytosis of CG membrane proteins is a prerequisite for efficient serial killing of CTLs and identify key events in this process. Copyright © 2016 by The American Association of Immunologists, Inc.

  8. Endocytosis of Integrin-Binding Human Picornaviruses

    Directory of Open Access Journals (Sweden)

    Pirjo Merilahti

    2012-01-01

    Full Text Available Picornaviruses that infect humans form one of the largest virus groups with almost three hundred virus types. They include significant enteroviral pathogens such as rhino-, polio-, echo-, and coxsackieviruses and human parechoviruses that cause wide range of disease symptoms. Despite the economic importance of picornaviruses, there are no antivirals. More than ten cellular receptors are known to participate in picornavirus infection, but experimental evidence of their role in cellular infection has been shown for only about twenty picornavirus types. Three enterovirus types and one parechovirus have experimentally been shown to bind and use integrin receptors in cellular infection. These include coxsackievirus A9 (CV-A9, echovirus 9, and human parechovirus 1 that are among the most common and epidemic human picornaviruses and bind to αV-integrins via RGD motif that resides on virus capsid. In contrast, echovirus 1 (E-1 has no RGD and uses integrin α2β1 as cellular receptor. Endocytosis of CV-A9 has recently been shown to occur via a novel Arf6- and dynamin-dependent pathways, while, contrary to collagen binding, E-1 binds inactive β1 integrin and enters via macropinocytosis. In this paper, we review what is known about receptors and endocytosis of integrin-binding human picornaviruses.

  9. Endocytosis of heat-denatured albumin by cultured rat Kupffer cells

    International Nuclear Information System (INIS)

    Brouwer, A.; Knook, D.L.

    1982-01-01

    Purified Kupffer cells were obtained by centrifugal elutriation of sinusoidal cells isolated by pronase treatment of the rat liver. The endocytosis of radioactively labeled heat-aggregated colloidal albumin (CA 125 I) was investigated in maintenance cultures of the purified Kupffer cells. The endocytic capacity of the cells was studied during 4 days of culture. Maximum uptake was observed after 24 hr of culture, with a gradual decline during the following days. When the uptake was measured after incubation with increasing concentrations of CA 125 I, a saturation effect was observed. This finding and the observed high rate of uptake are strong indications that receptor sites on the cell membrane are involved in the mechanism of endocytosis. The uptake of CA 125 I by Kupffer cells was inhibited by the metabolic inhibitors fluoride and antimycin A, indicating that endocytosis of CA 125 I is dependent on energy derived from both glycolysis and mitochondrial respiration. The mechanism of internalization may also require the action of microfilaments as well as intact microtubules, since both cytochalasin B and colchicine inhibited the uptake of CA 125 I. The intracellular degradation of CA 125 I by Kupffer cells was strongly inhibited by chloroquine but not by colchicine. The degradation of ingested CA 125 I occurred within the Kupffer cell lysosomes

  10. Protein C inhibits endocytosis of thrombin-thrombomodulin complexes in A549 lung cancer cells and human umbilical vein endothelial cells

    International Nuclear Information System (INIS)

    Maruyama, I.; Majerus, P.W.

    1987-01-01

    We investigated the effect of protein C on the endocytosis of thrombin-thrombomodulin complexes. We previously showed that exposure of umbilical vein endothelial cells to thrombin stimulated the internalization and degradation of thrombin. A similar internalization was stimulated by a monoclonal antithrombomodulin antibody. We have repeated these studies in the presence of protein C and found that endocytosis of 125 I-thrombin-thrombomodulin complexes, but not 125 I-antithrombomodulin-thrombomodulin complexes, is inhibited. Activated protein C did not inhibit endocytosis of thrombin-thrombomodulin complexes. Protein C inhibited both internalization and degradation of 125 I-thrombin and diisopropylphosphoryl (DIP) 125 I-thrombin in human lung cancer cells (A549). These effects were observed at protein C concentrations found in human plasma. Protein S had no effect on the inhibition of endocytosis of thrombin-thrombomodulin complexes by protein C. We propose that protein C may regulate the rate of endocytosis of thrombin-thrombomodulin complexes in vivo and thereby control the capacity for endothelium to activate protein C

  11. Clathrin to Lipid Raft-Endocytosis via Controlled Surface Chemistry and Efficient Perinuclear Targeting of Nanoparticle.

    Science.gov (United States)

    Chakraborty, Atanu; Jana, Nikhil R

    2015-09-17

    Nanoparticle interacts with live cells depending on their surface chemistry, enters into cell via endocytosis, and is commonly trafficked to an endosome/lysozome that restricts subcellular targeting options. Here we show that nanoparticle surface chemistry can be tuned to alter their cell uptake mechanism and subcellular trafficking. Quantum dot based nanoprobes of 20-30 nm hydrodynamic diameters have been synthesized with tunable surface charge (between +15 mV to -25 mV) and lipophilicity to influence their cellular uptake processes and subcellular trafficking. It is observed that cationic nanoprobe electrostatically interacts with cell membrane and enters into cell via clathrin-mediated endocytosis. At lower surface charge (between +10 mV to -10 mV), the electrostatic interaction with cell membrane becomes weaker, and additional lipid raft endocytosis is initiated. If a lipophilic functional group is introduced on a weakly anionic nanoparticle surface, the uptake mechanism shifts to predominant lipid raft-mediated endocytosis. In particular, the zwitterionic-lipophilic nanoprobe has the unique advantage as it weakly interacts with anionic cell membrane, migrates toward lipid rafts for interaction through lipophilic functional group, and induces lipid raft-mediated endocytosis. While predominate or partial clathrin-mediated entry traffics most of the nanoprobes to lysozome, predominate lipid raft-mediated entry traffics them to perinuclear region, particularly to the Golgi apparatus. This finding would guide in designing appropriate nanoprobe for subcellular targeting and delivery.

  12. Distinct Functions of Endophilin Isoforms in Synaptic Vesicle Endocytosis

    Directory of Open Access Journals (Sweden)

    Jifeng Zhang

    2015-01-01

    Full Text Available Endophilin isoforms perform distinct characteristics in their interactions with N-type Ca2+ channels and dynamin. However, precise functional differences for the endophilin isoforms on synaptic vesicle (SV endocytosis remain unknown. By coupling RNA interference and electrophysiological recording techniques in cultured rat hippocampal neurons, we investigated the functional differences of three isoforms of endophilin in SV endocytosis. The results showed that the amplitude of normalized evoked excitatory postsynaptic currents in endophilin1 knockdown neurons decreased significantly for both single train and multiple train stimulations. Similar results were found using endophilin2 knockdown neurons, whereas endophilin3 siRNA exhibited no change compared with control neurons. Endophilin1 and endophilin2 affected SV endocytosis, but the effect of endophilin1 and endophilin2 double knockdown was not different from that of either knockdown alone. This result suggested that endophilin1 and endophilin2 functioned together but not independently during SV endocytosis. Taken together, our results indicate that SV endocytosis is sustained by endophilin1 and endophilin2 isoforms, but not by endophilin3, in primary cultured hippocampal neurons.

  13. Non-Ligand-Induced Dimerization is Sufficient to Initiate the Signalling and Endocytosis of EGF Receptor

    Directory of Open Access Journals (Sweden)

    George Kourouniotis

    2016-07-01

    Full Text Available The binding of epidermal growth factor (EGF to EGF receptor (EGFR stimulates cell mitogenesis and survival through various signalling cascades. EGF also stimulates rapid EGFR endocytosis and its eventual degradation in lysosomes. The immediate events induced by ligand binding include receptor dimerization, activation of intrinsic tyrosine kinase and autophosphorylation. However, in spite of intensified efforts, the results regarding the roles of these events in EGFR signalling and internalization is still very controversial. In this study, we constructed a chimeric EGFR by replacing its extracellular domain with leucine zipper (LZ and tagged a green fluorescent protein (GFP at its C-terminus. We showed that the chimeric LZ-EGFR-GFP was constitutively dimerized. The LZ-EGFR-GFP dimer autophosphorylated each of its five well-defined C-terminal tyrosine residues as the ligand-induced EGFR dimer does. Phosphorylated LZ-EGFR-GFP was localized to both the plasma membrane and endosomes, suggesting it is capable of endocytosis. We also showed that LZ-EGFR-GFP activated major signalling proteins including Src homology collagen-like (Shc, extracellular signal-regulated kinase (ERK and Akt. Moreover, LZ-EGFR-GFP was able to stimulate cell proliferation. These results indicate that non-ligand induced dimerization is sufficient to activate EGFR and initiate cell signalling and EGFR endocytosis. We conclude that receptor dimerization is a critical event in EGF-induced cell signalling and EGFR endocytosis.

  14. Dynamic bio-adhesion of polymer nanoparticles on MDCK epithelial cells and its impact on bio-membranes, endocytosis and paracytosis.

    Science.gov (United States)

    He, Bing; Yuan, Lan; Dai, Wenbing; Gao, Wei; Zhang, Hua; Wang, Xueqing; Fang, Weigang; Zhang, Qiang

    2016-03-21

    Nowadays, concern about the use of nanotechnology for biomedical application is unprecedentedly increasing. In fact, nanosystems applied for various potential clinical uses always have to cross the primary biological barrier consisting of epithelial cells. However, little is really known currently in terms of the influence of the dynamic bio-adhesion of nanosystems on bio-membranes as well as on endocytosis and transcytosis. This was investigated here using polymer nanoparticles (PNs) and MDCK epithelial cells as the models. Firstly, the adhesion of PNs on cell membranes was found to be time-dependent with a shift of both location and dispersion pattern, from the lateral adhesion of mainly mono-dispersed PNs initially to the apical coverage of the PN aggregate later. Then, it was interesting to observe in this study that the dynamic bio-adhesion of PNs only affected their endocytosis but not their transcytosis. It was important to find that the endocytosis of PNs was not a constant process. A GM1 dependent CDE (caveolae dependent endocytosis) pathway was dominant in the preliminary stage, followed by the co-existence of a CME (clathrin-mediated endocytosis) pathway for the PN aggregate at a later stage, in accordance with the adhesion features of PNs, suggesting the modification of PN adhesion patterns on the endocytosis pathways. Next, the PN adhesion was noticed to affect the structure of cell junctions, via altering the extra- and intra-cellular calcium levels, leading to the enhanced paracellular transport of small molecules, but not favorably enough for the obviously increased passing of PNs themselves. Finally, FRAP and other techniques all demonstrated the obvious impact of PN adhesion on the membrane confirmation, independent of the adhesion location and time, which might lower the threshold for the internalization of PNs, even their aggregates. Generally, these findings confirm that the transport pathway mechanism of PNs through epithelial cells is rather

  15. Dynamic bio-adhesion of polymer nanoparticles on MDCK epithelial cells and its impact on bio-membranes, endocytosis and paracytosis

    Science.gov (United States)

    He, Bing; Yuan, Lan; Dai, Wenbing; Gao, Wei; Zhang, Hua; Wang, Xueqing; Fang, Weigang; Zhang, Qiang

    2016-03-01

    Nowadays, concern about the use of nanotechnology for biomedical application is unprecedentedly increasing. In fact, nanosystems applied for various potential clinical uses always have to cross the primary biological barrier consisting of epithelial cells. However, little is really known currently in terms of the influence of the dynamic bio-adhesion of nanosystems on bio-membranes as well as on endocytosis and transcytosis. This was investigated here using polymer nanoparticles (PNs) and MDCK epithelial cells as the models. Firstly, the adhesion of PNs on cell membranes was found to be time-dependent with a shift of both location and dispersion pattern, from the lateral adhesion of mainly mono-dispersed PNs initially to the apical coverage of the PN aggregate later. Then, it was interesting to observe in this study that the dynamic bio-adhesion of PNs only affected their endocytosis but not their transcytosis. It was important to find that the endocytosis of PNs was not a constant process. A GM1 dependent CDE (caveolae dependent endocytosis) pathway was dominant in the preliminary stage, followed by the co-existence of a CME (clathrin-mediated endocytosis) pathway for the PN aggregate at a later stage, in accordance with the adhesion features of PNs, suggesting the modification of PN adhesion patterns on the endocytosis pathways. Next, the PN adhesion was noticed to affect the structure of cell junctions, via altering the extra- and intra-cellular calcium levels, leading to the enhanced paracellular transport of small molecules, but not favorably enough for the obviously increased passing of PNs themselves. Finally, FRAP and other techniques all demonstrated the obvious impact of PN adhesion on the membrane confirmation, independent of the adhesion location and time, which might lower the threshold for the internalization of PNs, even their aggregates. Generally, these findings confirm that the transport pathway mechanism of PNs through epithelial cells is rather

  16. Adaptor protein complex 2-mediated endocytosis is crucial for male reproductive organ development in Arabidopsis.

    Science.gov (United States)

    Kim, Soo Youn; Xu, Zheng-Yi; Song, Kyungyoung; Kim, Dae Heon; Kang, Hyangju; Reichardt, Ilka; Sohn, Eun Ju; Friml, Jirí; Juergens, Gerd; Hwang, Inhwan

    2013-08-01

    Fertilization in flowering plants requires the temporal and spatial coordination of many developmental processes, including pollen production, anther dehiscence, ovule production, and pollen tube elongation. However, it remains elusive as to how this coordination occurs during reproduction. Here, we present evidence that endocytosis, involving heterotetrameric adaptor protein complex 2 (AP-2), plays a crucial role in fertilization. An Arabidopsis thaliana mutant ap2m displays multiple defects in pollen production and viability, as well as elongation of staminal filaments and pollen tubes, all of which are pivotal processes needed for fertilization. Of these abnormalities, the defects in elongation of staminal filaments and pollen tubes were partially rescued by exogenous auxin. Moreover, DR5rev:GFP (for green fluorescent protein) expression was greatly reduced in filaments and anthers in ap2m mutant plants. At the cellular level, ap2m mutants displayed defects in both endocytosis of N-(3-triethylammonium-propyl)-4-(4-diethylaminophenylhexatrienyl) pyridinium dibromide, a lypophilic dye used as an endocytosis marker, and polar localization of auxin-efflux carrier PIN FORMED2 (PIN2) in the stamen filaments. Moreover, these defects were phenocopied by treatment with Tyrphostin A23, an inhibitor of endocytosis. Based on these results, we propose that AP-2-dependent endocytosis plays a crucial role in coordinating the multiple developmental aspects of male reproductive organs by modulating cellular auxin level through the regulation of the amount and polarity of PINs.

  17. Endocytosis and exocytosis of nanoparticles in mammalian cells

    Science.gov (United States)

    Oh, Nuri; Park, Ji-Ho

    2014-01-01

    Engineered nanoparticles that can be injected into the human body hold tremendous potential to detect and treat complex diseases. Understanding of the endocytosis and exocytosis mechanisms of nanoparticles is essential for safe and efficient therapeutic application. In particular, exocytosis is of significance in the removal of nanoparticles with drugs and contrast agents from the body, while endocytosis is of great importance for the targeting of nanoparticles in disease sites. Here, we review the recent research on the endocytosis and exocytosis of functionalized nanoparticles based on various sizes, shapes, and surface chemistries. We believe that this review contributes to the design of safe nanoparticles that can efficiently enter and leave human cells and tissues. PMID:24872703

  18. Actin and Endocytosis in Budding Yeast

    Science.gov (United States)

    Goode, Bruce L.; Eskin, Julian A.; Wendland, Beverly

    2015-01-01

    Endocytosis, the process whereby the plasma membrane invaginates to form vesicles, is essential for bringing many substances into the cell and for membrane turnover. The mechanism driving clathrin-mediated endocytosis (CME) involves > 50 different protein components assembling at a single location on the plasma membrane in a temporally ordered and hierarchal pathway. These proteins perform precisely choreographed steps that promote receptor recognition and clustering, membrane remodeling, and force-generating actin-filament assembly and turnover to drive membrane invagination and vesicle scission. Many critical aspects of the CME mechanism are conserved from yeast to mammals and were first elucidated in yeast, demonstrating that it is a powerful system for studying endocytosis. In this review, we describe our current mechanistic understanding of each step in the process of yeast CME, and the essential roles played by actin polymerization at these sites, while providing a historical perspective of how the landscape has changed since the preceding version of the YeastBook was published 17 years ago (1997). Finally, we discuss the key unresolved issues and where future studies might be headed. PMID:25657349

  19. Receptor-Mediated Endocytosis and Brain Delivery of Therapeutic Biologics

    Directory of Open Access Journals (Sweden)

    Guangqing Xiao

    2013-01-01

    Full Text Available Transport of macromolecules across the blood-brain-barrier (BBB requires both specific and nonspecific interactions between macromolecules and proteins/receptors expressed on the luminal and/or the abluminal surfaces of the brain capillary endothelial cells. Endocytosis and transcytosis play important roles in the distribution of macromolecules. Due to the tight junction of BBB, brain delivery of traditional therapeutic proteins with large molecular weight is generally not possible. There are multiple pathways through which macromolecules can be taken up into cells through both specific and nonspecific interactions with proteins/receptors on the cell surface. This review is focused on the current knowledge of receptor-mediated endocytosis/transcytosis and brain delivery using the Angiopep-2-conjugated system and the molecular Trojan horses. In addition, the role of neonatal Fc receptor (FcRn in regulating the efflux of Immunoglobulin G (IgG from brain to blood, and approaches to improve the pharmacokinetics of therapeutic biologics by generating Fc fusion proteins, and increasing the pH dependent binding affinity between Fc and FcRn, are discussed.

  20. Endocytosis of ABCG2 drug transporter caused by binding of 5D3 antibody: trafficking mechanisms and intracellular fate.

    Science.gov (United States)

    Studzian, Maciej; Bartosz, Grzegorz; Pulaski, Lukasz

    2015-08-01

    ABCG2, a metabolite and xenobiotic transporter located at the plasma membrane (predominantly in barrier tissues and progenitor cells), undergoes a direct progressive endocytosis process from plasma membrane to intracellular compartments upon binding of 5D3 monoclonal antibody. This antibody is specific to an external epitope on the protein molecule and locks it in a discrete conformation within its activity cycle, presumably providing a structural trigger for the observed internalization phenomenon. Using routine and novel assays, we show that ABCG2 is endocytosed by a mixed mechanism: partially via a rapid, clathrin-dependent pathway and partially in a cholesterol-dependent, caveolin-independent manner. While the internalization process is entirely dynamin-dependent and converges initially at the early endosome, subsequent intracellular fate of ABCG2 is again twofold: endocytosis leads to only partial lysosomal degradation, while a significant fraction of the protein is retained in a post-endosomal compartment with the possibility of at least partial recycling back to the cell surface. This externally triggered, conformation-related trafficking pathway may serve as a general regulatory paradigm for membrane transporters, and its discovery was made possible thanks to consistent application of quantitative methods. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Neutron activation analysis of bulk samples from Chinese ancient porcelain to provenance research

    International Nuclear Information System (INIS)

    Jian Zhu; Wentao Hao; Jianming Zhen; Tongxiu Zhen; Glascock, M.D.

    2013-01-01

    Neutron activation analysis (NAA) is an important technique to determine the provenance of ancient ceramics. The most common technique used for preparing ancient samples for NAA is to grind them into a powder and then encapsulate them before neutron irradiation. Unfortunately, ceramic materials are typically very hard making it a challenge to grind them into a powder. In this study we utilize bulk porcelain samples cut from ancient shards. The bulk samples are irradiated by neutrons alongside samples that have been conventionally ground into a powder. The NAA for both the bulk samples and powders are compared and shown to provide equivalent information regarding their chemical composition. Also, the multivariate statistical have been employed to the analysis data for check the consistency. The findings suggest that NAA results are less dependent on the state of the porcelain sample, and thus bulk samples cut from shards may be used to effectively determine their provenance. (author)

  2. Critical current characteristics and history dependence in superconducting SmFeAsOF bulk

    International Nuclear Information System (INIS)

    Ni, B; Ge, J; Kiuchi, M; Otabe, E S; Gao, Z; Wang, L; Qi, Y; Zhang, X; Ma, Y

    2010-01-01

    The superconducting SmFeAsO 1-x F x (x=0.2) polycrystalline bulks were prepared by the powder-in-tube (PIT) method. The magnetic field and temperature dependences of critical current densities in the samples were investigated by resistive and ac inductive (Campbell's) methods. It was found that a fairly large shielding current density over 10 9 A/m 2 , which is considered to correspond to the local critical current density, flows locally with the perimeter size similar to the average grain size of the bulk samples, while an extremely low transport current density of about 10 5 A/m 2 corresponding to the global critical current density flows through the whole sample. Furthermore, a unique history dependence of global critical current density was observed, i.e., it shows a smaller value in the increasing-field process than that in the decreasing-field process. The history dependence of global critical current characteristic in our case can be ascribed to the existence of the weak-link property between the grains in SmFeAsO 1-x F x bulk.

  3. Effect of endocytosis inhibitors on Coxiella burnetii interaction with host cells

    International Nuclear Information System (INIS)

    Tujulin, E.; Macellaro, A.; Norlander, L.; Liliehoeoek, B.

    1998-01-01

    The obligate intracellular rickettsia Coxiella burnetii has previously been reported to reach the intra-vacuolar compartment of host cells by phagocytosis. With the aim to further examine the mechanisms of C. burnetii internalisation, macrophage monolayers were treated with well characterised inhibitors of endocytosis. The treatment with two general inhibitors, colchicine and methylamine, resulted in a pronounced dose-dependent decrease of radiolabelled phase II rickettsiae retained from the intracellular fraction. A third inhibitor used, amiloride, has been reported to reduce effectively clathrin-independent pinocytic pathways. The internalisation of C. burnetii was shown to be substantially reduced also by amiloride and the effect was dependent on its concentration. The passive role of C. burnetii in the internalisation was verified by using heat-killed C. burnetii. Host cells treated with either of the three inhibitors (amiloride, colchicine and methylamine) showed a similar reduction of intracellular C. burnetii after exposure to killed as weal as live organisms. The data presented indicate that different endocytic mechanisms, pinocytosis as well as phagocytosis, may mediate the uptake of C. burnetii by a host cell. Key words: Coxiella burnetii; internalisation; endocytosis (authors)

  4. Ultrasound Targeted Microbubble Destruction Stimulates Cellular Endocytosis in Facilitation of Adeno-Associated Virus Delivery

    Directory of Open Access Journals (Sweden)

    Lian-Fang Du

    2013-05-01

    Full Text Available The generally accepted mechanism for ultrasound targeted microbubble destruction (UTMD to enhance drug and gene delivery is through sonoporation. However, passive uptake of adeno-associated virus (AAV into cells following sonoporation does not adequately explain observations of enhanced transduction by UTMD. This study investigated alternative mechanisms of UTMD enhancement in AAV delivery. UTMD significantly enhanced transduction efficiency of AAV in a dose-dependent manner. UTMD stimulated a persistent uptake of AAV into the cytoplasm and nucleus. This phenomenon occurred over several hours, suggesting that some viral particles are endocytosed by cells rather than exclusively passing through pores created by sonoporation. Additionally, UTMD enhanced clathrin expression and accumulation at the plasma membrane suggesting greater clathrin-mediated endocytosis following UTMD. Transmission electron microscopy (TEM revealed that UTMD stimulated formation of clathrin-coated pits (CPs and uncoated pits (nCPs. Furthermore, inhibition of clathrin-mediated endocytosis partially blocked the enhancement of AAV uptake following UTMD. The results of this study implicate endocytosis as a mechanism that contributes to UTMD-enhanced AAV delivery.

  5. The translocation of fullerenic nanoparticles into lysosome via the pathway of clathrin-mediated endocytosis

    International Nuclear Information System (INIS)

    Li Wei; Chen Chunying; Ye Chang; Zhao Yuliang; Chen Zhen; Meng Huan; Gao Yuxi; Yuan Hui; Xing Genmei; Zhao Feng; Chai Zhifang; Wei Taotao; Zhang Xujia; Yang Fuyu; Lao Fang; Han Dong; Tang Xianhua; Zhang Yingge

    2008-01-01

    Manufactured fullerene nanoparticles easily enter into cells and hence have been rapidly developed for biomedical uses. However, it is generally unknown which route the nanoparticles undergo when crossing cell membranes and where they localize to the intracellular compartments. Herein we have used both microscopic imaging and biological techniques to explore the processes of [C 60 (C(COOH) 2 ) 2 ] n nanoparticles across cellular membranes and their intracellular translocation in 3T3 L1 and RH-35 living cells. The fullerene nanoparticles are quickly internalized by the cells and then routed to the cytoplasm with punctate localization. Upon entering the cell, they are synchronized to lysosome-like vesicles. The [C 60 (C(COOH) 2 ) 2 ] n nanoparticles entering cells are mainly via endocytosis with time-, temperature- and energy-dependent manners. The cellular uptake of [C 60 (C(COOH) 2 ) 2 ] n nanoparticles was found to be clathrin-mediated but not caveolae-mediated endocytosis. The endocytosis mechanism and the subcellular target location provide key information for the better understanding and predicting of the biomedical function of fullerene nanoparticles inside cells

  6. Single Event Resolution of Plant Plasma Membrane Protein Endocytosis by TIRF Microscopy.

    Science.gov (United States)

    Johnson, Alexander; Vert, Grégory

    2017-01-01

    Endocytosis is a key process in the internalization of extracellular materials and plasma membrane proteins, such as receptors and transporters, thereby controlling many aspects of cell signaling and cellular homeostasis. Endocytosis in plants has an essential role not only for basic cellular functions but also for growth and development, nutrient delivery, toxin avoidance, and pathogen defense. The precise mechanisms of endocytosis in plants remain quite elusive. The lack of direct visualization and examination of single events of endocytosis has greatly hampered our ability to precisely monitor the cell surface lifetime and the recruitment profile of proteins driving endocytosis or endocytosed cargos in plants. Here, we discuss the necessity to systematically implement total internal reflection fluorescence microcopy (TIRF) in the Plant Cell Biology community and present reliable protocols for high spatial and temporal imaging of endocytosis in plants using clathrin-mediated endocytosis as a test case, since it represents the major route for internalization of cell-surface proteins in plants. We developed a robust method to directly visualize cell surface proteins using TIRF microscopy combined to a high throughput, automated and unbiased analysis pipeline to determine the temporal recruitment profile of proteins to single sites of endocytosis, using the departure of clathrin as a physiological reference for scission. Using this 'departure assay', we assessed the recruitment of two different AP-2 subunits, alpha and mu, to the sites of endocytosis and found that AP2A1 was recruited in concert with clathrin, while AP2M was not. This validated approach therefore offers a powerful solution to better characterize the plant endocytic machinery and the dynamics of one's favorite cargo protein.

  7. Temperature dependence of bulk respiration of crop stands. Measurement and model fitting

    International Nuclear Information System (INIS)

    Tani, Takashi; Arai, Ryuji; Tako, Yasuhiro

    2007-01-01

    The objective of the present study was to examine whether the temperature dependence of respiration at a crop-stand scale could be directly represented by an Arrhenius function that was widely used for representing the temperature dependence of leaf respiration. We determined temperature dependences of bulk respiration of monospecific stands of rice and soybean within a range of the air temperature from 15 to 30degC using large closed chambers. Measured responses of respiration rates of the two stands were well fitted by the Arrhenius function (R 2 =0.99). In the existing model to assess the local radiological impact of the anthropogenic carbon-14, effects of the physical environmental factors on photosynthesis and respiration of crop stands are not taken into account for the calculation of the net amount of carbon per cultivation area in crops at harvest which is the crucial parameter for the estimation of the activity concentration of carbon-14 in crops. Our result indicates that the Arrhenius function is useful for incorporating the effect of the temperature on respiration of crop stands into the model which is expected to contribute to a more realistic estimate of the activity concentration of carbon-14 in crops. (author)

  8. Interrelated temperature dependence of bulk etch rate and track length saturation time in CR-39 detector

    International Nuclear Information System (INIS)

    Azooz, A.A.; Al-Jubbori, M.A.

    2013-01-01

    Highlights: • New empirical parameterization of CR-39 bulk etch rate. • Bulk etch rates measurements using two different methods give consistent results. • Temperature independence of track saturation length. • Two empirical relation between bulk etch rate and temperature are suggested. • Simple inverse relation between bulk etch rate and track saturation time. -- Abstract: Experimental measurements of the etching solution temperature dependence of bulk etch rate using two independent methods revealed a few interesting properties. It is found that while the track saturation length is independent of etching temperature, the etching time needed to reach saturation is strongly temperature-dependent. It is demonstrated that there is systematic simple inverse relation between track saturation time, and etching solution temperature. In addition, and although, the relation between the bulk etch rate and etching solution temperature can be reasonably described by a modified form of the Arrhenius equation, better fits can be obtained by another equation suggested in this work

  9. Internalization of titanium dioxide nanoparticles by glial cells is given at short times and is mainly mediated by actin reorganization-dependent endocytosis.

    Science.gov (United States)

    Huerta-García, Elizabeth; Márquez-Ramírez, Sandra Gissela; Ramos-Godinez, María Del Pilar; López-Saavedra, Alejandro; Herrera, Luis Alonso; Parra, Alberto; Alfaro-Moreno, Ernesto; Gómez, Erika Olivia; López-Marure, Rebeca

    2015-12-01

    Many nanoparticles (NPs) have toxic effects on multiple cell lines. This toxicity is assumed to be related to their accumulation within cells. However, the process of internalization of NPs has not yet been fully characterized. In this study, the cellular uptake, accumulation, and localization of titanium dioxide nanoparticles (TiO2 NPs) in rat (C6) and human (U373) glial cells were analyzed using time-lapse microscopy (TLM) and transmission electron microscopy (TEM). Cytochalasin D (Cyt-D) was used to evaluate whether the internalization process depends of actin reorganization. To determine whether the NP uptake is mediated by phagocytosis or macropinocytosis, nitroblue tetrazolium (NBT) reduction was measured and the 5-(N-ethyl-N-isopropyl)-amiloride was used. Expression of proteins involved with endocytosis and exocytosis such as caveolin-1 (Cav-1) and cysteine string proteins (CSPs) was also determined using flow cytometry. TiO2 NPs were taken up by both cell types, were bound to cellular membranes and were internalized at very short times after exposure (C6, 30 min; U373, 2h). During the uptake process, the formation of pseudopodia and intracellular vesicles was observed, indicating that this process was mediated by endocytosis. No specific localization of TiO2 NPs into particular organelles was found: in contrast, they were primarily localized into large vesicles in the cytoplasm. Internalization of TiO2 NPs was strongly inhibited by Cyt-D in both cells and by amiloride in U373 cells; besides, the observed endocytosis was not associated with NBT reduction in either cell type, indicating that macropinocytosis is the main process of internalization in U373 cells. In addition, increases in the expression of Cav-1 protein and CSPs were observed. In conclusion, glial cells are able to internalize TiO2 NPs by a constitutive endocytic mechanism which may be associated with their strong cytotoxic effect in these cells; therefore, TiO2 NPs internalization and their

  10. Cancer cell-selective, clathrin-mediated endocytosis of aptamer decorated nanoparticles

    Science.gov (United States)

    Engelberg, Shira; Modrejewski, Julia; Walter, Johanna G.; Livney, Yoav D.; Assaraf, Yehuda G.

    2018-01-01

    Lung cancer is the leading cause of cancer mortality worldwide, resulting in 88% deaths of all diagnosed patients. Hence, novel therapeutic modalities are urgently needed. Single-stranded oligonucleotide-based aptamers (APTs) are excellent ligands for tumor cell targeting. However, the molecular mechanisms underlying their internalization into living cells have been poorly studied. Towards the application of APTs for active drug targeting to cancer cells, we herein studied the mechanism underlying S15-APT internalization into human non-small cell lung cancer A549 cells. We thus delineated the mode of entry of a model nanomedical system based on quantum dots (QDs) decorated with S15-APTs as a selective targeting moiety for uptake by A549 cells. These APT-decorated QDs displayed selective binding to, and internalization by target A549 cells, but not by normal human bronchial epithelial BEAS2B, cervical carcinoma (HeLa) and colon adenocarcinoma CaCo-2 cells, hence demonstrating high specificity. Flow cytometric analysis revealed a remarkably low dissociation constant of S15-APTs-decorated QDs to A549 cells (Kd = 13.1 ± 1.6 nM). Through the systematic application of a series of established inhibitors of known mechanisms of endocytosis, we show that the uptake of S15-APTs proceeds via a classical clathrin-dependent receptor-mediated endocytosis. This cancer cell-selective mode of entry could possibly be used in the future to evade plasma membrane-localized multidrug resistance efflux pumps, thereby overcoming an important mechanism of cancer multidrug resistance. PMID:29765515

  11. Phosphorylation of the Usher syndrome 1G protein SANS controls Magi2-mediated endocytosis.

    Science.gov (United States)

    Bauß, Katharina; Knapp, Barbara; Jores, Pia; Roepman, Ronald; Kremer, Hannie; Wijk, Erwin V; Märker, Tina; Wolfrum, Uwe

    2014-08-01

    The human Usher syndrome (USH) is a complex ciliopathy with at least 12 chromosomal loci assigned to three clinical subtypes, USH1-3. The heterogeneous USH proteins are organized into protein networks. Here, we identified Magi2 (membrane-associated guanylate kinase inverted-2) as a new component of the USH protein interactome, binding to the multifunctional scaffold protein SANS (USH1G). We showed that the SANS-Magi2 complex assembly is regulated by the phosphorylation of an internal PDZ-binding motif in the sterile alpha motif domain of SANS by the protein kinase CK2. We affirmed Magi2's role in receptor-mediated, clathrin-dependent endocytosis and showed that phosphorylated SANS tightly regulates Magi2-mediated endocytosis. Specific depletions by RNAi revealed that SANS and Magi2-mediated endocytosis regulates aspects of ciliogenesis. Furthermore, we demonstrated the localization of the SANS-Magi2 complex in the periciliary membrane complex facing the ciliary pocket of retinal photoreceptor cells in situ. Our data suggest that endocytotic processes may not only contribute to photoreceptor cell homeostasis but also counterbalance the periciliary membrane delivery accompanying the exocytosis processes for the cargo vesicle delivery. In USH1G patients, mutations in SANS eliminate Magi2 binding and thereby deregulate endocytosis, lead to defective ciliary transport modules and ultimately disrupt photoreceptor cell function inducing retinal degeneration. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  12. Adaptor Protein Complex 2–Mediated Endocytosis Is Crucial for Male Reproductive Organ Development in Arabidopsis[W

    Science.gov (United States)

    Kim, Soo Youn; Xu, Zheng-Yi; Song, Kyungyoung; Kim, Dae Heon; Kang, Hyangju; Reichardt, Ilka; Sohn, Eun Ju; Friml, Jiří; Juergens, Gerd; Hwang, Inhwan

    2013-01-01

    Fertilization in flowering plants requires the temporal and spatial coordination of many developmental processes, including pollen production, anther dehiscence, ovule production, and pollen tube elongation. However, it remains elusive as to how this coordination occurs during reproduction. Here, we present evidence that endocytosis, involving heterotetrameric adaptor protein complex 2 (AP-2), plays a crucial role in fertilization. An Arabidopsis thaliana mutant ap2m displays multiple defects in pollen production and viability, as well as elongation of staminal filaments and pollen tubes, all of which are pivotal processes needed for fertilization. Of these abnormalities, the defects in elongation of staminal filaments and pollen tubes were partially rescued by exogenous auxin. Moreover, DR5rev:GFP (for green fluorescent protein) expression was greatly reduced in filaments and anthers in ap2m mutant plants. At the cellular level, ap2m mutants displayed defects in both endocytosis of N-(3-triethylammonium-propyl)-4-(4-diethylaminophenylhexatrienyl) pyridinium dibromide, a lypophilic dye used as an endocytosis marker, and polar localization of auxin-efflux carrier PIN FORMED2 (PIN2) in the stamen filaments. Moreover, these defects were phenocopied by treatment with Tyrphostin A23, an inhibitor of endocytosis. Based on these results, we propose that AP-2–dependent endocytosis plays a crucial role in coordinating the multiple developmental aspects of male reproductive organs by modulating cellular auxin level through the regulation of the amount and polarity of PINs. PMID:23975898

  13. Contributions of herpes simplex virus type 1 envelope proteins to entry by endocytosis

    Science.gov (United States)

    Herpes simplex virus (HSV) proteins specifically required for endocytic entry but not direct penetration have not been identified. HSVs deleted of gE, gG, gI, gJ, gM, UL45, or Us9 entered cells via either pH-dependent or pH-independent endocytosis and were inactivated by mildly acidic pH. Thus, the ...

  14. ApoER2 expression increases Aβ production while decreasing Amyloid Precursor Protein (APP endocytosis: Possible role in the partitioning of APP into lipid rafts and in the regulation of γ-secretase activity

    Directory of Open Access Journals (Sweden)

    Bu Guojun

    2007-07-01

    Full Text Available Abstract Background The generation of the amyloid-β peptide (Aβ through the proteolytic processing of the amyloid precursor protein (APP is a central event in the pathogenesis of Alzheimer's disease (AD. Recent studies highlight APP endocytosis and localization to lipid rafts as important events favoring amyloidogenic processing. However, the precise mechanisms underlying these events are poorly understood. ApoER2 is a member of the low density lipoprotein receptor (LDL-R family exhibiting slow endocytosis rate and a significant association with lipid rafts. Despite the important neurophysiological roles described for ApoER2, little is known regarding how ApoER2 regulates APP trafficking and processing. Results Here, we demonstrate that ApoER2 physically interacts and co-localizes with APP. Remarkably, we found that ApoER2 increases cell surface APP levels and APP association with lipid rafts. The increase of cell surface APP requires the presence of ApoER2 cytoplasmic domain and is a result of decreased APP internalization rate. Unexpectedly, ApoER2 expression correlated with a significant increase in Aβ production and reduced levels of APP-CTFs. The increased Aβ production was dependent on the integrity of the NPxY endocytosis motif of ApoER2. We also found that expression of ApoER2 increased APP association with lipid rafts and increased γ-secretase activity, both of which might contribute to increased Aβ production. Conclusion These findings show that ApoER2 negatively affects APP internalization. However, ApoER2 expression stimulates Aβ production by shifting the proportion of APP from the non-rafts to the raft membrane domains, thereby promoting β-secretase and γ-secretase mediated amyloidogenic processing and also by incrementing the activity of γ-secretase.

  15. Electric field dependence of the electron mobility in bulk wurtzite ZnO

    Indian Academy of Sciences (India)

    Electric field dependence of the electron mobility in bulk wurtzite ZnO. K ALFARAMAWI ... tion to ultraviolet light emitters, gas sensors, surface acoustic wave devices and ..... Dorkel J M and Leturcq P H 1981 Solid-State Electron. 24 8211.

  16. Emission-energy dependence of ultrafast P-emission decay in ZnO from bulk to nanofilm

    International Nuclear Information System (INIS)

    Wakaiki, Shuji; Ichida, Hideki; Bamba, Motoaki; Kawase, Toshiki; Kawakami, Masaki; Mizoguchi, Kohji; Kim, DaeGwi; Nakayama, Masaaki; Kanematsu, Yasuo

    2014-01-01

    We have performed time-resolved photoluminescence (PL) spectroscopy for ZnO thin films with thicknesses of 90, 460, and 2800 nm under intense excitation condition. We clearly observed the P emission due to inelastic exciton–exciton scattering. It was found that, in the 460- and 2800-nm thick samples, the decay time of the P emission considerably depends on the detection energy inversely proportional to the group velocity of the polariton in a bulk crystal with each factor of proportionality. In contrast, the energy dependence is less remarkable in the 90-nm thick sample. The decay times are basically shortened with a decrease in the film thickness. The thickness dependence of the P-emission-decay profiles is explained by considering the crossover from the polariton modes in the 2800-nm thick sample (bulk-like film) to the exciton-/photon-like modes in the 90-nm thick sample (nanofilm). - Highlights: • We clearly observed the P-PL dynamics due to inelastic exciton–exciton scattering. • The P-PL decay times are basically shortened with a decrease in the film thickness. • The P-PL decay time depends on the detection energy in the bulk-like sample. • The energy dependence of the P-PL decay time almost disappears in the 90-nm sample. • The thickness dependence is explained by the crossover between exciton and photon

  17. Emission-energy dependence of ultrafast P-emission decay in ZnO from bulk to nanofilm

    Energy Technology Data Exchange (ETDEWEB)

    Wakaiki, Shuji, E-mail: s.wakaiki@mls.eng.osaka-u.ac.jp [Department of Material and Life Science, Division of Advanced Science and Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan); Ichida, Hideki [Department of Material and Life Science, Division of Advanced Science and Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan); Science and Technology Entrepreneurship Laboratory, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan); Bamba, Motoaki [Department of Physics, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043 (Japan); Kawase, Toshiki; Kawakami, Masaki [Department of Applied Physics, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585 (Japan); Mizoguchi, Kohji [Department of Physical Science, Graduate School of Science, Osaka Prefecture University, 1-1 Gakuen, Naka-ku, Sakai, Osaka 599-8531 (Japan); Kim, DaeGwi; Nakayama, Masaaki [Department of Applied Physics, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585 (Japan); Kanematsu, Yasuo [Department of Material and Life Science, Division of Advanced Science and Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan); Science and Technology Entrepreneurship Laboratory, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan)

    2014-08-01

    We have performed time-resolved photoluminescence (PL) spectroscopy for ZnO thin films with thicknesses of 90, 460, and 2800 nm under intense excitation condition. We clearly observed the P emission due to inelastic exciton–exciton scattering. It was found that, in the 460- and 2800-nm thick samples, the decay time of the P emission considerably depends on the detection energy inversely proportional to the group velocity of the polariton in a bulk crystal with each factor of proportionality. In contrast, the energy dependence is less remarkable in the 90-nm thick sample. The decay times are basically shortened with a decrease in the film thickness. The thickness dependence of the P-emission-decay profiles is explained by considering the crossover from the polariton modes in the 2800-nm thick sample (bulk-like film) to the exciton-/photon-like modes in the 90-nm thick sample (nanofilm). - Highlights: • We clearly observed the P-PL dynamics due to inelastic exciton–exciton scattering. • The P-PL decay times are basically shortened with a decrease in the film thickness. • The P-PL decay time depends on the detection energy in the bulk-like sample. • The energy dependence of the P-PL decay time almost disappears in the 90-nm sample. • The thickness dependence is explained by the crossover between exciton and photon.

  18. Mobility of tethering factor EEA1 on endosomes is decreased upon stimulation of EGF receptor endocytosis in HeLa cells

    Energy Technology Data Exchange (ETDEWEB)

    Kosheverova, Vera V., E-mail: kosheverova_vera@incras.ru [Institute of Cytology of RAS, 4, Tikhoretsky Ave, St. Petersburg, 194064 (Russian Federation); Kamentseva, Rimma S., E-mail: rkamentseva@yandex.ru [Institute of Cytology of RAS, 4, Tikhoretsky Ave, St. Petersburg, 194064 (Russian Federation); St. Petersburg State University, 7-9, Universitetskaya nab, St. Petersburg, 199034 (Russian Federation); Gonchar, Ilya V., E-mail: ample@mail.ru [Institute of Cytology of RAS, 4, Tikhoretsky Ave, St. Petersburg, 194064 (Russian Federation); Kharchenko, Marianna V., E-mail: mariannakharchenko@gmail.com [Institute of Cytology of RAS, 4, Tikhoretsky Ave, St. Petersburg, 194064 (Russian Federation); Kornilova, Elena S., E-mail: lenkor@mail.cytspb.rssi.ru [Institute of Cytology of RAS, 4, Tikhoretsky Ave, St. Petersburg, 194064 (Russian Federation); St. Petersburg State University, 7-9, Universitetskaya nab, St. Petersburg, 199034 (Russian Federation); Department of Medical Physics, Peter the Great St. Petersburg Polytechnic University, 29, Polytechnicheskaya, St.Petersburg, 195251 (Russian Federation)

    2016-04-22

    Tethering factor EEA1, mediating homotypic fusion of early endosomes, was shown to be localized in membrane-bound state both in serum-deprived and stimulated for EGF receptor endocytosis cells. However, it is not known whether dynamics behavior of EEA1 is affected by EGF stimulation. We investigated EEA1 cytosol-to-membrane exchange rate in interphase HeLa cells by FRAP analysis. The data obtained fitted two-states binding model, with the bulk of membrane-associated EEA1 protein represented by the mobile fraction both in serum-starved and EGF-stimulated cells. Fast recovery state had similar half-times in the two cases: about 1.6 s and 2.8 s, respectively. However, the recovery half-time of slowly cycled EEA1 fraction significantly increased in EGF-stimulated comparing to serum-starved cells (from 21 to 99 s). We suppose that the retardation of EEA1 fluorescence recovery upon EGF-stimulation may be due to the increase of activated Rab5 on endosomal membranes, the growth of the number of tethering events between EEA1-positive vesicles and their clustering. - Highlights: • EEA1 mobility was compared in serum-starved and EGF-stimulated interphase HeLa cells. • FRAP analysis revealed fast and slow components of EEA1 recovery in both cases. • Stimulation of EGFR endocytosis did not affect fast EEA1 turnover. • EGF stimulation significantly increased half-time of slowly exchanged EEA1 fraction.

  19. Mobility of tethering factor EEA1 on endosomes is decreased upon stimulation of EGF receptor endocytosis in HeLa cells

    International Nuclear Information System (INIS)

    Kosheverova, Vera V.; Kamentseva, Rimma S.; Gonchar, Ilya V.; Kharchenko, Marianna V.; Kornilova, Elena S.

    2016-01-01

    Tethering factor EEA1, mediating homotypic fusion of early endosomes, was shown to be localized in membrane-bound state both in serum-deprived and stimulated for EGF receptor endocytosis cells. However, it is not known whether dynamics behavior of EEA1 is affected by EGF stimulation. We investigated EEA1 cytosol-to-membrane exchange rate in interphase HeLa cells by FRAP analysis. The data obtained fitted two-states binding model, with the bulk of membrane-associated EEA1 protein represented by the mobile fraction both in serum-starved and EGF-stimulated cells. Fast recovery state had similar half-times in the two cases: about 1.6 s and 2.8 s, respectively. However, the recovery half-time of slowly cycled EEA1 fraction significantly increased in EGF-stimulated comparing to serum-starved cells (from 21 to 99 s). We suppose that the retardation of EEA1 fluorescence recovery upon EGF-stimulation may be due to the increase of activated Rab5 on endosomal membranes, the growth of the number of tethering events between EEA1-positive vesicles and their clustering. - Highlights: • EEA1 mobility was compared in serum-starved and EGF-stimulated interphase HeLa cells. • FRAP analysis revealed fast and slow components of EEA1 recovery in both cases. • Stimulation of EGFR endocytosis did not affect fast EEA1 turnover. • EGF stimulation significantly increased half-time of slowly exchanged EEA1 fraction.

  20. Correlated Fluorescence-Atomic Force Microscopy Studies of the Clathrin Mediated Endocytosis in SKMEL Cells

    Science.gov (United States)

    Smith, Steve; Hor, Amy; Luu, Anh; Kang, Lin; Scott, Brandon; Bailey, Elizabeth; Hoppe, Adam

    Clathrin-mediated endocytosis is one of the central pathways for cargo transport into cells, and plays a major role in the maintenance of cellular functions, such as intercellular signaling, nutrient intake, and turnover of plasma membrane in cells. The clathrin-mediated endocytosis process involves invagination and formation of clathrin-coated vesicles. However, the biophysical mechanisms of vesicle formation are still debated. We investigate clathrin vesicle formation mechanisms through the utilization of tapping-mode atomic force microscopy for high resolution topographical imaging in neutral buffer solution of unroofed cells exposing the inner membrane, combined with fluorescence imaging to definitively label intracellular constituents with specific fluorescent fusion proteins (actin filaments labeled with green phalloidin-antibody and clathrin coated vesicles with the fusion protein Tq2) in SKMEL (Human Melanoma) cells. Results from our work are compared against dynamical polarized total internal fluorescence (TIRF), super-resolution photo-activated localization microscopy (PALM) and transmission electron microscopy (TEM) to draw conclusions regarding the prominent model of vesicle formation in clathrin-mediated endocytosis. Funding provided by NSF MPS/DMR/BMAT award # 1206908.

  1. Epidermal Growth Factor Enhances Cellular Uptake of Polystyrene Nanoparticles by Clathrin-Mediated Endocytosis.

    Science.gov (United States)

    Phuc, Le Thi Minh; Taniguchi, Akiyoshi

    2017-06-19

    The interaction between nanoparticles and cells has been studied extensively, but most research has focused on the effect of various nanoparticle characteristics, such as size, morphology, and surface charge, on the cellular uptake of nanoparticles. In contrast, there have been very few studies to assess the influence of cellular factors, such as growth factor responses, on the cellular uptake efficiency of nanoparticles. The aim of this study was to clarify the effects of epidermal growth factor (EGF) on the uptake efficiency of polystyrene nanoparticles (PS NPs) by A431 cells, a human carcinoma epithelial cell line. The results showed that EGF enhanced the uptake efficiency of A431 cells for PS NPs. In addition, inhibition and localization studies of PS NPs and EGF receptors (EGFRs) indicated that cellular uptake of PS NPs is related to the binding of EGF-EGFR complex and PS NPs. Different pathways are used to enter the cells depending on the presence or absence of EGF. In the presence of EGF, cellular uptake of PS NPs is via clathrin-mediated endocytosis, whereas, in the absence of EGF, uptake of PS NPs does not involve clathrin-mediated endocytosis. Our findings indicate that EGF enhances cellular uptake of PS NPs by clathrin-mediated endocytosis. This result could be important for developing safe nanoparticles and their safe use in medical applications.

  2. Epidermal Growth Factor Enhances Cellular Uptake of Polystyrene Nanoparticles by Clathrin-Mediated Endocytosis

    Directory of Open Access Journals (Sweden)

    Le Thi Minh Phuc

    2017-06-01

    Full Text Available The interaction between nanoparticles and cells has been studied extensively, but most research has focused on the effect of various nanoparticle characteristics, such as size, morphology, and surface charge, on the cellular uptake of nanoparticles. In contrast, there have been very few studies to assess the influence of cellular factors, such as growth factor responses, on the cellular uptake efficiency of nanoparticles. The aim of this study was to clarify the effects of epidermal growth factor (EGF on the uptake efficiency of polystyrene nanoparticles (PS NPs by A431 cells, a human carcinoma epithelial cell line. The results showed that EGF enhanced the uptake efficiency of A431 cells for PS NPs. In addition, inhibition and localization studies of PS NPs and EGF receptors (EGFRs indicated that cellular uptake of PS NPs is related to the binding of EGF–EGFR complex and PS NPs. Different pathways are used to enter the cells depending on the presence or absence of EGF. In the presence of EGF, cellular uptake of PS NPs is via clathrin-mediated endocytosis, whereas, in the absence of EGF, uptake of PS NPs does not involve clathrin-mediated endocytosis. Our findings indicate that EGF enhances cellular uptake of PS NPs by clathrin-mediated endocytosis. This result could be important for developing safe nanoparticles and their safe use in medical applications.

  3. Genetically encoded pH sensor for tracking surface proteins through endocytosis.

    Science.gov (United States)

    Grover, Anmol; Schmidt, Brigitte F; Salter, Russell D; Watkins, Simon C; Waggoner, Alan S; Bruchez, Marcel P

    2012-05-14

    Traffic cam: a tandem dye prepared from a FRET acceptor and a fluorogenic donor functions as a cell surface ratiometric pH indicator, which upon internalization serves to follow protein trafficking during endocytosis. This sensor was used to analyze agonist-dependent internalization of β(2)-adrenergic receptors. It was also used as a surrogate antigen to reveal direct surface-to-endosome antigen transfer between dendritic cells (not shown). Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Ultrasound Microbubble Treatment Enhances Clathrin-Mediated Endocytosis and Fluid-Phase Uptake through Distinct Mechanisms.

    Directory of Open Access Journals (Sweden)

    Farnaz Fekri

    Full Text Available Drug delivery to tumors is limited by several factors, including drug permeability of the target cell plasma membrane. Ultrasound in combination with microbubbles (USMB is a promising strategy to overcome these limitations. USMB treatment elicits enhanced cellular uptake of materials such as drugs, in part as a result of sheer stress and formation of transient membrane pores. Pores formed upon USMB treatment are rapidly resealed, suggesting that other processes such as enhanced endocytosis may contribute to the enhanced material uptake by cells upon USMB treatment. How USMB regulates endocytic processes remains incompletely understood. Cells constitutively utilize several distinct mechanisms of endocytosis, including clathrin-mediated endocytosis (CME for the internalization of receptor-bound macromolecules such as Transferrin Receptor (TfR, and distinct mechanism(s that mediate the majority of fluid-phase endocytosis. Tracking the abundance of TfR on the cell surface and the internalization of its ligand transferrin revealed that USMB acutely enhances the rate of CME. Total internal reflection fluorescence microscopy experiments revealed that USMB treatment altered the assembly of clathrin-coated pits, the basic structural units of CME. In addition, the rate of fluid-phase endocytosis was enhanced, but with delayed onset upon USMB treatment relative to the enhancement of CME, suggesting that the two processes are distinctly regulated by USMB. Indeed, vacuolin-1 or desipramine treatment prevented the enhancement of CME but not of fluid phase endocytosis upon USMB, suggesting that lysosome exocytosis and acid sphingomyelinase, respectively, are required for the regulation of CME but not fluid phase endocytosis upon USMB treatment. These results indicate that USMB enhances both CME and fluid phase endocytosis through distinct signaling mechanisms, and suggest that strategies for potentiating the enhancement of endocytosis upon USMB treatment may

  5. DRP1-Dependent Endocytosis is Essential for Polar Localization and Boron-Induced Degradation of the Borate Transporter BOR1 in Arabidopsis thaliana.

    Science.gov (United States)

    Yoshinari, Akira; Fujimoto, Masaru; Ueda, Takashi; Inada, Noriko; Naito, Satoshi; Takano, Junpei

    2016-09-01

    Boron (B) is essential for plants but toxic in excess. The borate efflux transporter BOR1 is expressed in various root cells and localized to the inner/stele-side domain of the plasma membrane (PM) under low-B conditions. BOR1 is rapidly degraded through endocytosis upon sufficient B supply. The polar localization and degradation of BOR1 are considered important for efficient B translocation and avoidance of B toxicity, respectively. In this study, we first analyzed the subcellular localization of BOR1 in roots, cotyledons and hypocotyls, and revealed a polar localization in various cell types. We also found that the inner polarity of BOR1 is established after completion of cytokinesis in the root meristem. Moreover, variable-angle epifluorescence microscopy visualized BOR1-green fluorescent protein (GFP) as particles in the PM with significant lateral movements but in restricted areas. Importantly, a portion of BOR1-GFP particles co-localized with DYNAMIN-RELATED PROTEIN 1A (DRP1A), which is involved in scission of the clathrin-coated vesicles, and they disappeared together from the PM. To examine the contribution of DRP1A-mediated endocytosis to BOR1 localization and degradation, we developed an inducible expression system of the DRP1A K47A variant. The DRP1A variant prolonged the residence time of clathrin on the PM and inhibited endocytosis of membrane lipids. The dominant-negative DRP1A blocked endocytosis of BOR1 and disturbed its polar localization and B-induced degradation. Our results provided insight into the endocytic mechanisms that modulate the subcellular localization and abundance of a mineral transporter for nutrient homeostasis in plant cells. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  6. Podocytic PKC-alpha is regulated in murine and human diabetes and mediates nephrin endocytosis.

    Directory of Open Access Journals (Sweden)

    Irini Tossidou

    Full Text Available BACKGROUND: Microalbuminuria is an early lesion during the development of diabetic nephropathy. The loss of high molecular weight proteins in the urine is usually associated with decreased expression of slit diaphragm proteins. Nephrin, is the major component of the glomerular slit diaphragm and loss of nephrin has been well described in rodent models of experimental diabetes as well as in human diabetic nephropathy. METHODOLOGY/PRINCIPAL FINDINGS: In this manuscript we analyzed the role of PKC-alpha (PKCalpha on endocytosis of nephrin in podocytes. We found that treatment of diabetic mice with a PKCalpha-inhibitor (GO6976 leads to preserved nephrin expression and reduced proteinuria. In vitro, we found that high glucose stimulation would induce PKCalpha protein expression in murine and human podocytes. We can demonstrate that PKCalpha mediates nephrin endocytosis in podocytes and that overexpression of PKCalpha leads to an augmented endocytosis response. After PKC-activation, we demonstrate an inducible association of PKCalpha, PICK1 and nephrin in podocytes. Moreover, we can demonstrate a strong induction of PKCalpha in podocytes of patients with diabetic nephropathy. CONCLUSIONS/SIGNIFICANCE: We therefore conclude that activation of PKCalpha is a pathomechanistic key event during the development of diabetic nephropathy. PKCalpha is involved in reduction of nephrin surface expression and therefore PKCalpha inhibition might be a novel target molecule for anti-proteinuric therapy.

  7. Scavenger receptor-mediated endocytosis by sinusoidal cells in rat bone marrow

    International Nuclear Information System (INIS)

    Geoffroy, J.S.

    1987-01-01

    Endocytosis of serum albumin by sinusoidal endothelial cells in rat bone marrow was investigated initially at the ultrastructural level with subsequent biochemical investigation of the specificity mediating this event. Bovine serum albumin adsorbed to 20nm colloidal gold particles (AuBSA) was chosen as the electron microscopic probe. Morphological data strongly suggested that a receptor was involved in uptake of AuBSA. Confirmation of receptor involvement in the uptake of AuBSA by marrow sinusoidal endothelial cells was achieved utilizing an in situ isolated hind limb perfusion protocol in conjunction with unlabeled, radiolabeled, and radio-/colloidal gold labeled probes. The major findings of competition and saturation experiments were: (1) endocytosis of AuBSA was mediated by a receptor for modified/treated serum albumin; (2) endocytosis of formaldehyde-treated serum albumin was mediated by a binding site which may be the same or closely related to the site responsible for the uptake of AuBSA; and (3) endocytosis of native untreated albumin was not mediated by receptor and probably represents fluid-phase pinocitosis

  8. A Global Analysis of Kinase Function in Candida albicans Hyphal Morphogenesis Reveals a Role for the Endocytosis Regulator Akl1.

    Science.gov (United States)

    Bar-Yosef, Hagit; Gildor, Tsvia; Ramírez-Zavala, Bernardo; Schmauch, Christian; Weissman, Ziva; Pinsky, Mariel; Naddaf, Rawi; Morschhäuser, Joachim; Arkowitz, Robert A; Kornitzer, Daniel

    2018-01-01

    The human pathogenic fungus Candida albicans can switch between yeast and hyphal morphologies as a function of environmental conditions and cellular physiology. The yeast-to-hyphae morphogenetic switch is activated by well-established, kinase-based signal transduction pathways that are induced by extracellular stimuli. In order to identify possible inhibitory pathways of the yeast-to-hyphae transition, we interrogated a collection of C. albicans protein kinases and phosphatases ectopically expressed under the regulation of the TETon promoter. Proportionately more phosphatases than kinases were identified that inhibited hyphal morphogenesis, consistent with the known role of protein phosphorylation in hyphal induction. Among the kinases, we identified AKL1 as a gene that significantly suppressed hyphal morphogenesis in serum. Akl1 specifically affected hyphal elongation rather than initiation: overexpression of AKL1 repressed hyphal growth, and deletion of AKL1 resulted in acceleration of the rate of hyphal elongation. Akl1 suppressed fluid-phase endocytosis, probably via Pan1, a putative clathrin-mediated endocytosis scaffolding protein. In the absence of Akl1, the Pan1 patches were delocalized from the sub-apical region, and fluid-phase endocytosis was intensified. These results underscore the requirement of an active endocytic pathway for hyphal morphogenesis. Furthermore, these results suggest that under standard conditions, endocytosis is rate-limiting for hyphal elongation.

  9. Endocytosis of a functionally enhanced GFP-tagged transferrin receptor in CHO cells.

    Directory of Open Access Journals (Sweden)

    Qi He

    Full Text Available The endocytosis of transferrin receptor (TfR has served as a model to study the receptor-targeted cargo delivery system for cancer therapy for many years. To accurately evaluate and optically measure this TfR targeting delivery in vitro, a CHO cell line with enhanced green fluorescent protein (EGFP-tagged human TfR was established. A chimera of the hTfR and EGFP was engineered by fusing EGFP to the amino terminus of hTfR. Data were provided to demonstrate that hTfR-EGFP chimera was predominantly localized on the plasma membrane with some intracellular fluorescent structures on CHO cells and the EGFP moiety did not affect the endocytosis property of hTfR. Receptor internalization occurred similarly to that of HepG2 cells expressing wild-type hTfR. The internalization percentage of this chimeric receptor was about 81 ± 3% of wild type. Time-dependent co-localization of hTfR-EGFP and PE-conjugated anti-hTfR mAb in living cells demonstrated the trafficking of mAb-receptor complexes through the endosomes followed by segregation of part of the mAb and receptor at the late stages of endocytosis. The CHO-hTfR cells preferentially took up anti-hTfR mAb conjugated nanoparticles. This CHO-hTfR cell line makes it feasible for accurate evaluation and visualization of intracellular trafficking of therapeutic agents conjugated with transferrin or Abs targeting the hTfRs.

  10. Role of receptor-mediated endocytosis in the antiangiogenic effects of human T lymphoblastic cell-derived microparticles.

    Science.gov (United States)

    Yang, Chun; Xiong, Wei; Qiu, Qian; Shao, Zhuo; Shao, Zuo; Hamel, David; Tahiri, Houda; Leclair, Grégoire; Lachapelle, Pierre; Chemtob, Sylvain; Hardy, Pierre

    2012-04-15

    Microparticles possess therapeutic potential regarding angiogenesis. We have demonstrated the contribution of apoptotic human CEM T lymphocyte-derived microparticles (LMPs) as inhibitors of angiogenic responses in animal models of inflammation and tumor growth. In the present study, we characterized the antivascular endothelial growth factor (VEGF) effects of LMPs on pathological angiogenesis in an animal model of oxygen-induced retinopathy and explored the role of receptor-mediated endocytosis in the effects of LMPs on human retinal endothelial cells (HRECs). LMPs dramatically inhibited cell growth of HRECs, suppressed VEGF-induced cell migration in vitro experiments, and attenuated VEGF-induced retinal vascular leakage in vivo. Intravitreal injections of fluorescently labeled LMPs revealed accumulation of LMPs in retinal tissue, with more than 60% reductions of the vascular density in retinas of rats with oxygen-induced neovascularization. LMP uptake experiments demonstrated that the interaction between LMPs and HRECs is dependent on temperature. In addition, endocytosis is partially dependent on extracellular calcium. RNAi-mediated knockdown of low-density lipoprotein receptor (LDLR) reduced the uptake of LMPs and attenuated the inhibitory effects of LMPs on VEGF-A protein expression and HRECs cell growth. Intravitreal injection of lentivirus-mediated RNA interference reduced LDLR protein expression in retina by 53% and significantly blocked the antiangiogenic effects of LMPs on pathological vascularization. In summary, the potent antiangiogenic LMPs lead to a significant reduction of pathological retinal angiogenesis through modulation of VEGF signaling, whereas LDLR-mediated endocytosis plays a partial, but pivotal, role in the uptake of LMPs in HRECs.

  11. Effect of flow on endothelial endocytosis of nanocarriers targeted to ICAM-1.

    Science.gov (United States)

    Bhowmick, Tridib; Berk, Erik; Cui, Xiumin; Muzykantov, Vladimir R; Muro, Silvia

    2012-02-10

    Delivery of drugs into the endothelium by nanocarriers targeted to endothelial determinants may improve treatment of vascular maladies. This is the case for intercellular adhesion molecule 1 (ICAM-1), a glycoprotein overexpressed on endothelial cells (ECs) in many pathologies. ICAM-1-targeted nanocarriers bind to and are internalized by ECs via a non-classical pathway, CAM-mediated endocytosis. In this work we studied the effects of endothelial adaptation to physiological flow on the endocytosis of model polymer nanocarriers targeted to ICAM-1 (anti-ICAM/NCs, ~180 nm diameter). Culturing established endothelial-like cells (EAhy926 cells) and primary human umbilical vein ECs (HUVECs) under 4 dyn/cm(2) laminar shear stress for 24 h resulted in flow adaptation: cell elongation and formation of actin stress fibers aligned to the flow direction. Fluorescence microscopy showed that flow-adapted cells internalized anti-ICAM/NCs under flow, although at slower rate versus non flow-adapted cells under static incubation (~35% reduction). Uptake was inhibited by amiloride, whereas marginally affected by filipin and cadaverine, implicating that CAM-endocytosis accounts for anti-ICAM/NC uptake under flow. Internalization under flow was more modestly affected by inhibiting protein kinase C, which regulates actin remodeling during CAM-endocytosis. Actin recruitment to stress fibers that maintain the cell shape under flow may delay uptake of anti-ICAM/NCs under this condition by interfering with actin reorganization needed for CAM-endocytosis. Electron microscopy revealed somewhat slow, yet effective endocytosis of anti-ICAM/NCs by pulmonary endothelium after i.v. injection in mice, similar to that of flow-adapted cell cultures: ~40% (30 min) and 80% (3 h) internalization. Similar to cell culture data, uptake was slightly faster in capillaries with lower shear stress. Further, LPS treatment accelerated internalization of anti-ICAM/NCs in mice. Therefore, regulation of endocytosis

  12. Cooperative endocytosis of the endosomal SNARE protein syntaxin-8 and the potassium channel TASK-1

    Science.gov (United States)

    Renigunta, Vijay; Fischer, Thomas; Zuzarte, Marylou; Kling, Stefan; Zou, Xinle; Siebert, Kai; Limberg, Maren M.; Rinné, Susanne; Decher, Niels; Schlichthörl, Günter; Daut, Jürgen

    2014-01-01

    The endosomal SNARE protein syntaxin-8 interacts with the acid-sensitive potassium channel TASK-1. The functional relevance of this interaction was studied by heterologous expression of these proteins (and mutants thereof) in Xenopus oocytes and in mammalian cell lines. Coexpression of syntaxin-8 caused a fourfold reduction in TASK-1 current, a corresponding reduction in the expression of TASK-1 at the cell surface, and a marked increase in the rate of endocytosis of the channel. TASK-1 and syntaxin-8 colocalized in the early endosomal compartment, as indicated by the endosomal markers 2xFYVE and rab5. The stimulatory effect of the SNARE protein on the endocytosis of the channel was abolished when both an endocytosis signal in TASK-1 and an endocytosis signal in syntaxin-8 were mutated. A syntaxin-8 mutant that cannot assemble with other SNARE proteins had virtually the same effect as wild-type syntaxin-8. Total internal reflection fluorescence microscopy showed formation and endocytosis of vesicles containing fluorescence-tagged clathrin, TASK-1, and/or syntaxin-8. Our results suggest that the unassembled form of syntaxin-8 and the potassium channel TASK-1 are internalized via clathrin-mediated endocytosis in a cooperative manner. This implies that syntaxin-8 regulates the endocytosis of TASK-1. Our study supports the idea that endosomal SNARE proteins can have functions unrelated to membrane fusion. PMID:24743596

  13. PKC-Dependent GlyT1 Ubiquitination Occurs Independent of Phosphorylation: Inespecificity in Lysine Selection for Ubiquitination.

    Directory of Open Access Journals (Sweden)

    Susana P Barrera

    Full Text Available Neurotransmitter transporter ubiquitination is emerging as the main mechanism for endocytosis and sorting of cargo into lysosomes. In this study, we demonstrate PKC-dependent ubiquitination of three different isoforms of the glycine transporter 1 (GlyT1. Incubation of cells expressing transporter with the PKC activator phorbol ester induced a dramatic, time-dependent increase in GlyT1 ubiquitination, followed by accumulation of GlyT1 in EEA1 positive early endosomes. This occurred via a mechanism that was abolished by inhibition of PKC. GlyT1 endocytosis was confirmed in both retinal sections and primary cultures of mouse amacrine neurons. Replacement of only all lysines in the N-and C-termini to arginines prevented ubiquitination and endocytosis, displaying redundancy in the mechanism of ubiquitination. Interestingly, a 40-50% reduction in glycine uptake was detected in phorbol-ester stimulated cells expressing the WT-GlyT1, whereas no significant change was for the mutant protein, demonstrating that endocytosis participates in the reduction of uptake. Consistent with previous findings for the dopamine transporter DAT, ubiquitination of GlyT1 tails functions as sorting signal to deliver transporter into the lysosome and removal of ubiquitination sites dramatically attenuated the rate of GlyT1 degradation. Finally, we showed for the first time that PKC-dependent GlyT1 phosphorylation was not affected by removal of ubiquitination sites, suggesting separate PKC-dependent signaling events for these posttranslational modifications.

  14. Entry of porcine reproductive and respiratory syndrome virus into porcine alveolar macrophages via receptor-mediated endocytosis.

    Science.gov (United States)

    Nauwynck, H J; Duan, X; Favoreel, H W; Van Oostveldt, P; Pensaert, M B

    1999-02-01

    Porcine alveolar macrophages (AMphi) are the dominant cell type that supports the replication of porcine reproductive and respiratory syndrome virus (PRRSV) in vivo and in vitro. In order to determine the characteristics of the virus-receptor interaction, the attachment of PRRSV to cells was examined by using biotinylated virus in a series of flow cytometric assays. PRRSV bound specifically to AMphi in a dose-dependent manner. Binding of PRRSV to AMphi increased gradually and reached a maximum within 60 min at 4 degrees C. By confocal microscopy, it was shown that different degrees of PRRSV binding exist and that entry is by endocytosis. Virus uptake in vesicles is a clathrin-dependent process, as it was blocked by the addition of cytochalasin D and co-localization of PRRSV and clathrin was found. Furthermore, by the use of two weak bases, NH4Cl and chloroquine, it was demonstrated that PRRSV uses a low pH-dependent entry pathway. In the presence of these reagents, input virions accumulated in large vacuoles, indicating that uncoating was prevented. These results indicate that PRRSV entry into AMphi involves attachment to a specific virus receptor(s) followed by a process of endocytosis, by which virions are taken into the cell within vesicles by a clathrin-dependent pathway. A subsequent drop in pH is required for proper virus replication.

  15. Shiga Toxin—A Model for Glycolipid-Dependent and Lectin-Driven Endocytosis

    Directory of Open Access Journals (Sweden)

    Ludger Johannes

    2017-10-01

    Full Text Available The cellular entry of the bacterial Shiga toxin and the related verotoxins has been scrutinized in quite some detail. This is due to their importance as a threat to human health. At the same time, the study of Shiga toxin has allowed the discovery of novel molecular mechanisms that also apply to the intracellular trafficking of endogenous proteins at the plasma membrane and in the endosomal system. In this review, the individual steps that lead to Shiga toxin uptake into cells will first be presented from a purely mechanistic perspective. Membrane-biological concepts will be highlighted that are often still poorly explored, such as fluctuation force-driven clustering, clathrin-independent membrane curvature generation, friction-driven scission, and retrograde sorting on early endosomes. It will then be explored whether and how these also apply to other pathogens, pathogenic factors, and cellular proteins. The molecular nature of Shiga toxin as a carbohydrate-binding protein and that of its cellular receptor as a glycosylated raft lipid will be an underlying theme in this discussion. It will thereby be illustrated how the study of Shiga toxin has led to the proposal of the GlycoLipid-Lectin (GL-Lect hypothesis on the generation of endocytic pits in processes of clathrin-independent endocytosis.

  16. Caveolin-1 and CDC42 mediated endocytosis of silica-coated iron oxide nanoparticles in HeLa cells

    Directory of Open Access Journals (Sweden)

    Nils Bohmer

    2015-01-01

    Full Text Available Nanomedicine is a rapidly growing field in nanotechnology, which has great potential in the development of new therapies for numerous diseases. For example iron oxide nanoparticles are in clinical use already in the thermotherapy of brain cancer. Although it has been shown, that tumor cells take up these particles in vitro, little is known about the internalization routes. Understanding of the underlying uptake mechanisms would be very useful for faster and precise development of nanoparticles for clinical applications. This study aims at the identification of key proteins, which are crucial for the active uptake of iron oxide nanoparticles by HeLa cells (human cervical cancer as a model cell line. Cells were transfected with specific siRNAs against Caveolin-1, Dynamin 2, Flotillin-1, Clathrin, PIP5Kα and CDC42. Knockdown of Caveolin-1 reduces endocytosis of superparamagnetic iron oxide nanoparticles (SPIONs and silica-coated iron oxide nanoparticles (SCIONs between 23 and 41%, depending on the surface characteristics of the nanoparticles and the experimental design. Knockdown of CDC42 showed a 46% decrease of the internalization of PEGylated SPIONs within 24 h incubation time. Knockdown of Dynamin 2, Flotillin-1, Clathrin and PIP5Kα caused no or only minor effects. Hence endocytosis in HeLa cells of iron oxide nanoparticles, used in this study, is mainly mediated by Caveolin-1 and CDC42. It is shown here for the first time, which proteins of the endocytotic pathway mediate the endocytosis of silica-coated iron oxide nanoparticles in HeLa cells in vitro. In future studies more experiments should be carried out with different cell lines and other well-defined nanoparticle species to elucidate possible general principles.

  17. Hierarchical classification strategy for Phenotype extraction from epidermal growth factor receptor endocytosis screening

    NARCIS (Netherlands)

    L. Cao (Lu); M. Graauw (Marjo de); K. Yan (Kuan); L.C.J. Winkel (Leah C.J.); F.J. Verbeek (Fons)

    2016-01-01

    textabstractBackground: Endocytosis is regarded as a mechanism of attenuating the epidermal growth factor receptor (EGFR) signaling and of receptor degradation. There is increasing evidence becoming available showing that breast cancer progression is associated with a defect in EGFR endocytosis. In

  18. Mathematical modeling of white adipocyte exocytosis predicts adiponectin secretion and quantifies the rates of vesicle exo- and endocytosis.

    Science.gov (United States)

    Brännmark, Cecilia; Lövfors, William; Komai, Ali M; Axelsson, Tom; El Hachmane, Mickaël F; Musovic, Saliha; Paul, Alexandra; Nyman, Elin; Olofsson, Charlotta S

    2017-12-08

    Adiponectin is a hormone secreted from white adipocytes and takes part in the regulation of several metabolic processes. Although the pathophysiological importance of adiponectin has been thoroughly investigated, the mechanisms controlling its release are only partly understood. We have recently shown that adiponectin is secreted via regulated exocytosis of adiponectin-containing vesicles, that adiponectin exocytosis is stimulated by cAMP-dependent mechanisms, and that Ca 2+ and ATP augment the cAMP-triggered secretion. However, much remains to be discovered regarding the molecular and cellular regulation of adiponectin release. Here, we have used mathematical modeling to extract detailed information contained within our previously obtained high-resolution patch-clamp time-resolved capacitance recordings to produce the first model of adiponectin exocytosis/secretion that combines all mechanistic knowledge deduced from electrophysiological experimental series. This model demonstrates that our previous understanding of the role of intracellular ATP in the control of adiponectin exocytosis needs to be revised to include an additional ATP-dependent step. Validation of the model by introduction of data of secreted adiponectin yielded a very close resemblance between the simulations and experimental results. Moreover, we could show that Ca 2+ -dependent adiponectin endocytosis contributes to the measured capacitance signal, and we were able to predict the contribution of endocytosis to the measured exocytotic rate under different experimental conditions. In conclusion, using mathematical modeling of published and newly generated data, we have obtained estimates of adiponectin exo- and endocytosis rates, and we have predicted adiponectin secretion. We believe that our model should have multiple applications in the study of metabolic processes and hormonal control thereof. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. A Global Analysis of Kinase Function in Candida albicans Hyphal Morphogenesis Reveals a Role for the Endocytosis Regulator Akl1

    Directory of Open Access Journals (Sweden)

    Hagit Bar-Yosef

    2018-02-01

    Full Text Available The human pathogenic fungus Candida albicans can switch between yeast and hyphal morphologies as a function of environmental conditions and cellular physiology. The yeast-to-hyphae morphogenetic switch is activated by well-established, kinase-based signal transduction pathways that are induced by extracellular stimuli. In order to identify possible inhibitory pathways of the yeast-to-hyphae transition, we interrogated a collection of C. albicans protein kinases and phosphatases ectopically expressed under the regulation of the TETon promoter. Proportionately more phosphatases than kinases were identified that inhibited hyphal morphogenesis, consistent with the known role of protein phosphorylation in hyphal induction. Among the kinases, we identified AKL1 as a gene that significantly suppressed hyphal morphogenesis in serum. Akl1 specifically affected hyphal elongation rather than initiation: overexpression of AKL1 repressed hyphal growth, and deletion of AKL1 resulted in acceleration of the rate of hyphal elongation. Akl1 suppressed fluid-phase endocytosis, probably via Pan1, a putative clathrin-mediated endocytosis scaffolding protein. In the absence of Akl1, the Pan1 patches were delocalized from the sub-apical region, and fluid-phase endocytosis was intensified. These results underscore the requirement of an active endocytic pathway for hyphal morphogenesis. Furthermore, these results suggest that under standard conditions, endocytosis is rate-limiting for hyphal elongation.

  20. Quantification of cytosolic interactions identifies Ede1 oligomers as key organizers of endocytosis.

    Science.gov (United States)

    Boeke, Dominik; Trautmann, Susanne; Meurer, Matthias; Wachsmuth, Malte; Godlee, Camilla; Knop, Michael; Kaksonen, Marko

    2014-11-03

    Clathrin-mediated endocytosis is a highly conserved intracellular trafficking pathway that depends on dynamic protein-protein interactions between up to 60 different proteins. However, little is known about the spatio-temporal regulation of these interactions. Using fluorescence (cross)-correlation spectroscopy in yeast, we tested 41 previously reported interactions in vivo and found 16 to exist in the cytoplasm. These detected cytoplasmic interactions included the self-interaction of Ede1, homolog of mammalian Eps15. Ede1 is the crucial scaffold for the organization of the early stages of endocytosis. We show that oligomerization of Ede1 through its central coiled coil domain is necessary for its localization to the endocytic site and we link the oligomerization of Ede1 to its function in locally concentrating endocytic adaptors and organizing the endocytic machinery. Our study sheds light on the importance of the regulation of protein-protein interactions in the cytoplasm for the assembly of the endocytic machinery in vivo. © 2014 The Authors. Published under the terms of the CC BY 4.0 license.

  1. Hypothesis review: are clathrin-mediated endocytosis and clathrin-dependent membrane and protein trafficking core pathophysiological processes in schizophrenia and bipolar disorder?

    LENUS (Irish Health Repository)

    2012-02-01

    Clathrin-mediated endocytosis (CME) is the best-characterized mechanism governing cellular membrane and protein trafficking. In this hypothesis review, we integrate recent evidence implicating CME and related cellular trafficking mechanisms in the pathophysiology of psychotic disorders such as schizophrenia and bipolar disorder. The evidence includes proteomic and genomic findings implicating proteins and genes of the clathrin interactome. Additionally, several important candidate genes for schizophrenia, such as dysbindin, are involved in processes closely linked to CME and membrane trafficking. We discuss that key aspects of psychosis neuropathology such as synaptic dysfunction, white matter changes and aberrant neurodevelopment are all influenced by clathrin-dependent processes, and that other cellular trafficking mechanisms previously linked to psychoses interact with the clathrin interactome in important ways. Furthermore, many antipsychotic drugs have been shown to affect clathrin-interacting proteins. We propose that the targeted pharmacological manipulation of the clathrin interactome may offer fruitful opportunities for novel treatments of schizophrenia.Molecular Psychiatry advance online publication, 11 October 2011; doi:10.1038\\/mp.2011.123.

  2. [INHIBITORS OF MAP-KINASE PATHWAY U0126 AND PD98059 DIFFERENTLY AFFECT ORGANIZATION OF TUBULIN CYTOSKELETON AFTER STIMULATION OF EGF RECEPTOR ENDOCYTOSIS].

    Science.gov (United States)

    Zlobina, M V; Steblyanko, Yu Yu; Shklyaeva, M A; Kharchenko, V V; Salova, A V; Kornilova, E S

    2015-01-01

    To confirm the hypothesis about the involvement of EGF-stimulated MAP-kinase ERK1/2 in the regulation of microtubule (MT) system, the influence of two widely used ERK1/2 inhibitors, U0126 and PD98059, on the organization of tubulin cytoskeleton in interphase HeLa cells during EGF receptor endocytosis has been investigated. We have found that addition of U0126 or PD98059 to not-stimulated with EGF ells for 30 min has no effect on radially organized MT system. However, in the case of U0126 addition before EGF endocytosis stimulation, the number of MT per cell decreased within 15 min after such stimulation and was followed by complete MT depolymerization by 60-90 min. Stimulation of EGF endocytosis in the presence of PD98059 resulted only in insignificant depolymerization of MT and it could be detected mainly from their minus-ends. At the same time, MT regions close to plasma membrane became stabilized, which was proved by increase in tubulin acetylation level. This situation was characteristic for all period of the experiment. It has been also found that the inhibitors affect endocytosis dynamics of EGF-receptor complexes. Quantitative analysis demonstrated that the stimulation of endocytosis in the presence of U0126 generated a greater number of endosomes compared to control cells, and their number did not change significantly during the experiment. All these endosomes were localized peripherally. Effect of PD98059 resulted in the formation of lower number of endosomes that in control, but they demonstrated very slow clusterization despite the presence of some intact MT. Both inhibitors decreased EGFR colocolization with early endosomal marker EEA1, which indicated a delay in endosome fusions and maturation. The inhibitors were also shown to affect differently phospho-ERK 1 and 2 forms: U0126 completely inhibited phospho-ERK1 and 2, white, in the presence of PD98059, the two ERK forms demonstrated sharp transient activation in 15 min after stimulation, but only

  3. Transcytosis of F4 fimbriae by villous and dome epithelia in F4-receptor positive pigs supports importance of receptor-dependent endocytosis in oral immunization strategies.

    Science.gov (United States)

    Snoeck, Veerle; Van den Broeck, Wim; De Colvenaer, Veerle; Verdonck, Frank; Goddeeris, Bruno; Cox, Eric

    2008-07-15

    Very few antigens have been described that induce an intestinal immunity when given orally. Our laboratory demonstrated that oral administration of isolated F4 (K88) fimbriae of Escherichia coli to F4-receptor positive (F4R(+)) pigs induces protective mucosal immunity against challenge infection. However, presence of F4-receptors (F4R) on villous enterocytes is a prerequisite for inducing the immune response, as no F4-specific antibody-secreting cells (ASC) can be induced in F4R(-) pigs. In this study, the in vivo binding of isolated F4 fimbriae (F4) to the gut epithelium was examined in F4R(+) and F4R(-) pigs. It was further investigated whether binding of F4 to the F4R results in endocytosis in and translocation across the gut epithelium using microscopy. F4 did not adhere to the intestinal epithelium of F4R(-) pigs, whereas it strongly adhered to the villous epithelium and the follicle-associated epithelium (FAE) of the jejunum and ileum of F4R(+) pigs. Following binding to F4R, F4 was endocytosed by villous enterocytes, follicle-associated enterocytes and M cells. Transcytosis of F4 across the epithelium resulted in the appearance of F4 in the lamina propria and dome region of the jejunal and ileal PP. This is the first study showing transcytosis of fimbriae across the gut epithelium. This receptor-dependent transcytosis can explain the success of F4 fimbriae as oral immunogen for inducing protective immunity in F4R(+) pigs strengthening the importance of receptor-dependent endocytosis and translocation in oral vaccine strategies. Further identification of the receptor responsible for this transport is in progress.

  4. Quantitative Measurement of GPCR Endocytosis via Pulse-Chase Covalent Labeling.

    Directory of Open Access Journals (Sweden)

    Hidetoshi Kumagai

    Full Text Available G protein-coupled receptors (GPCRs play a critical role in many physiological systems and represent one of the largest families of signal-transducing receptors. The number of GPCRs at the cell surface regulates cellular responsiveness to their cognate ligands, and the number of GPCRs, in turn, is dynamically controlled by receptor endocytosis. Recent studies have demonstrated that GPCR endocytosis, in addition to affecting receptor desensitization and resensitization, contributes to acute G protein-mediated signaling. Thus, endocytic GPCR behavior has a significant impact on various aspects of physiology. In this study, we developed a novel GPCR internalization assay to facilitate characterization of endocytic GPCR behavior. We genetically engineered chimeric GPCRs by fusing HaloTag (a catalytically inactive derivative of a bacterial hydrolase to the N-terminal end of the receptor (HT-GPCR. HaloTag has the ability to form a stable covalent bond with synthetic HaloTag ligands that contain fluorophores or a high-affinity handle (such as biotin and the HaloTag reactive linker. We selectively labeled HT-GPCRs at the cell surface with a HaloTag PEG ligand, and this pulse-chase covalent labeling allowed us to directly monitor the relative number of internalized GPCRs after agonist stimulation. Because the endocytic activities of GPCR ligands are not necessarily correlated with their agonistic activities, applying this novel methodology to orphan GPCRs, or even to already characterized GPCRs, will increase the likelihood of identifying currently unknown ligands that have been missed by conventional pharmacological assays.

  5. Nanomechanics of magnetically driven cellular endocytosis

    Czech Academy of Sciences Publication Activity Database

    Zablotskyy, Vitaliy A.; Lunov, O.; Dejneka, Alexandr; Jastrabík, Lubomír; Polyakova, T.; Syrovets, T.; Simmet, T.

    2011-01-01

    Roč. 99, č. 18 (2011), 183701/1-183701/3 ISSN 0003-6951 R&D Projects: GA AV ČR KAN301370701; GA MŠk(CZ) 1M06002 Institutional research plan: CEZ:AV0Z10100522 Keywords : magnetically controled endocytosis * cell membranes * iron oxide nanoparticles Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 3.844, year: 2011

  6. Electric field dependence of the electron mobility in bulk wurtzite ZnO

    Indian Academy of Sciences (India)

    ZnO) material is studied. The low-field electron mobility is calculated as a function of doping concentration and lattice temperature. The results show that above nearly 50 K the electrical conduction is governed by activation through the bulk ...

  7. Distinct functions of Crumbs regulating slit diaphragms and endocytosis in Drosophila nephrocytes.

    Science.gov (United States)

    Hochapfel, Florian; Denk, Lucia; Mendl, Gudrun; Schulze, Ulf; Maaßen, Christine; Zaytseva, Yulia; Pavenstädt, Hermann; Weide, Thomas; Rachel, Reinhard; Witzgall, Ralph; Krahn, Michael P

    2017-12-01

    Mammalian podocytes, the key determinants of the kidney's filtration barrier, differentiate from columnar epithelial cells and several key determinants of apical-basal polarity in the conventional epithelia have been shown to regulate podocyte morphogenesis and function. However, little is known about the role of Crumbs, a conserved polarity regulator in many epithelia, for slit-diaphragm formation and podocyte function. In this study, we used Drosophila nephrocytes as model system for mammalian podocytes and identified a conserved function of Crumbs proteins for cellular morphogenesis, nephrocyte diaphragm assembly/maintenance, and endocytosis. Nephrocyte-specific knock-down of Crumbs results in disturbed nephrocyte diaphragm assembly/maintenance and decreased endocytosis, which can be rescued by Drosophila Crumbs as well as human Crumbs2 and Crumbs3, which were both expressed in human podocytes. In contrast to the extracellular domain, which facilitates nephrocyte diaphragm assembly/maintenance, the intracellular FERM-interaction motif of Crumbs is essential for regulating endocytosis. Moreover, Moesin, which binds to the FERM-binding domain of Crumbs, is essential for efficient endocytosis. Thus, we describe here a new mechanism of nephrocyte development and function, which is likely to be conserved in mammalian podocytes.

  8. Chronic ethanol consumption in rats produces opioid antinociceptive tolerance through inhibition of mu opioid receptor endocytosis.

    Directory of Open Access Journals (Sweden)

    Li He

    Full Text Available It is well known that the mu-opioid receptor (MOR plays an important role in the rewarding properties of ethanol. However, it is less clear how chronic ethanol consumption affects MOR signaling. Here, we demonstrate that rats with prolonged voluntary ethanol consumption develop antinociceptive tolerance to opioids. Signaling through the MOR is controlled at many levels, including via the process of endocytosis. Importantly, agonists at the MOR that promote receptor endocytosis, such as the endogenous peptides enkephalin and β-endorphin, show a reduced propensity to promote antinociceptive tolerance than do agonists, like morphine, which do not promote receptor endocytosis. These observations led us to examine whether chronic ethanol consumption produced opioid tolerance by interfering with MOR endocytosis. Indeed, here we show that chronic ethanol consumption inhibits the endocytosis of MOR in response to opioid peptide. This loss of endocytosis was accompanied by a dramatic decrease in G protein coupled receptor kinase 2 (GRK2 protein levels after chronic drinking, suggesting that loss of this component of the trafficking machinery could be a mechanism by which endocytosis is lost. We also found that MOR coupling to G-protein was decreased in ethanol-drinking rats, providing a functional explanation for loss of opioid antinociception. Together, these results suggest that chronic ethanol drinking alters the ability of MOR to endocytose in response to opioid peptides, and consequently, promotes tolerance to the effects of opioids.

  9. Temperature dependence of bulk modulus and second-order elastic constants

    International Nuclear Information System (INIS)

    Singh, P.P.; Kumar, Munish

    2004-01-01

    A simple theoretical model is developed to investigate the temperature dependence of the bulk modulus and second order elastic constants. The method is based on the two different approaches viz. (i) the theory of thermal expansivity formulated by Suzuki, based on the Mie-Gruneisen equation of state, (ii) the theory of high-pressure-high-temperature equation of state formulated by Kumar, based on thermodynamic analysis. The results obtained for a number of crystals viz. NaCl, KCl, MgO and (Mg, Fe) 2 SiO 4 are discussed and compared with the experimental data. It is concluded that the Kumar formulation is far better that the Suzuki theory of thermal expansivity

  10. Temperature dependence of the interband critical points of bulk Ge and strained Ge on Si

    Science.gov (United States)

    Fernando, Nalin S.; Nunley, T. Nathan; Ghosh, Ayana; Nelson, Cayla M.; Cooke, Jacqueline A.; Medina, Amber A.; Zollner, Stefan; Xu, Chi; Menendez, Jose; Kouvetakis, John

    2017-11-01

    Epitaxial Ge layers on a Si substrate experience a tensile biaxial stress due to the difference between the thermal expansion coefficients of the Ge epilayer and the Si substrate, which can be measured using asymmetric X-ray diffraction reciprocal space maps. This stress depends on temperature and affects the band structure, interband critical points, and optical spectra. This manuscripts reports careful measurements of the temperature dependence of the dielectric function and the interband critical point parameters of bulk Ge and Ge epilayers on Si using spectroscopic ellipsometry from 80 to 780 K and from 0.8 to 6.5 eV. The authors find a temperature-dependent redshift of the E1 and E1 + Δ1 critical points in Ge on Si (relative to bulk Ge). This redshift can be described well with a model based on thermal expansion coefficients, continuum elasticity theory, and the deformation potential theory for interband transitions. The interband transitions leading to E0‧ and E2 critical points have lower symmetry and therefore are not affected by the stress.

  11. The immunological synapse: a focal point for endocytosis and exocytosis.

    Science.gov (United States)

    Griffiths, Gillian M; Tsun, Andy; Stinchcombe, Jane C

    2010-05-03

    There are many different cells in the immune system. To mount an effective immune response, they need to communicate with each other. One way in which this is done is by the formation of immunological synapses between cells. Recent developments show that the immune synapse serves as a focal point for exocytosis and endocytosis, directed by centrosomal docking at the plasma membrane. In this respect, formation of the immunological synapse bears striking similarities to cilia formation and cytokinesis. These intriguing observations suggest that the centrosome may play a conserved role in designating a specialized area of membrane for localized endocytosis and exocytosis.

  12. Ubiquitin-Mediated Regulation of Endocytosis by Proteins of the Arrestin Family

    Directory of Open Access Journals (Sweden)

    Michel Becuwe

    2012-01-01

    Full Text Available In metazoans, proteins of the arrestin family are key players of G-protein-coupled receptors (GPCRS signaling and trafficking. Following stimulation, activated receptors are phosphorylated, thus allowing the binding of arrestins and hence an “arrest” of receptor signaling. Arrestins act by uncoupling receptors from G proteins and contribute to the recruitment of endocytic proteins, such as clathrin, to direct receptor trafficking into the endocytic pathway. Arrestins also serve as adaptor proteins by promoting the recruitment of ubiquitin ligases and participate in the agonist-induced ubiquitylation of receptors, known to have impact on their subcellular localization and stability. Recently, the arrestin family has expanded following the discovery of arrestin-related proteins in other eukaryotes such as yeasts or fungi. Surprisingly, most of these proteins are also involved in the ubiquitylation and endocytosis of plasma membrane proteins, thus suggesting that the role of arrestins as ubiquitin ligase adaptors is at the core of these proteins' functions. Importantly, arrestins are themselves ubiquitylated, and this modification is crucial for their function. In this paper, we discuss recent data on the intricate connections between arrestins and the ubiquitin pathway in the control of endocytosis.

  13. Comparison of platelet activation through hinge vs bulk flow in mechanical heart valves

    Science.gov (United States)

    Hedayat, Mohammadali; Borazjani, Iman

    2017-11-01

    Bileaflet mechanical heart valves increase the risk of thrombus formation in patients which is believed to be initiated by platelet activation. Platelets can be activated by the elevated shear stresses in the bulk flow during the systole phase or the flow through the hinge during the diastole. However, the importance of platelet activation by the bulk flow vs the hinge in MHVs has yet to be studied. Here, we investigate the contribution of each of the above mechanisms to the activation of platelets in MHs by performing simulation of the flow through a 25mm St. Jude Medical valve placed in a straight aorta. Two different gap sizes (250 and 150 micrometer) are used in this study. The simulations are done using a sharp interface curvilinear immersed boundary method along with a strong-coupling algorithm for FSI solver on overset grids. The platelet activation through the hinge for different gap sizes is compared to the activation in the bulk flow using two platelet activation models to ensure the consistency of the results. Our results for all gap sizes using different activation models show that the integration of platelet activation caused by the bulk flow is several times higher in comparison to the activation through the hinge. This work is supported by the American Heart Association Grant 13SDG17220022, and the computational resources were partly provided by Center for Computational Research (CCR) at University at Buffalo.

  14. Orbital-dependent Rashba coupling in bulk BiTeCl and BiTeI

    KAUST Repository

    Zhu, Zhiyong

    2013-02-06

    By all-electron ab initio calculations, the layered polar semiconductor BiTeCl is shown to host giant bulk Rashba spin splitting, similar to the recently reported compound BiTeI. In both materials, the standard Rashba–Bychkov model is no longer applicable, because of huge band extrema shifts even in the absence of spin–orbit coupling and a strong momentum dependence of the Rashba coupling constant (αR). By assuming αR to be orbital dependent, a phenomenological extension of the Rashba–Bychkov model is proposed which explains the splitting behavior of states with small in-plane momentum.

  15. Orbital-dependent Rashba coupling in bulk BiTeCl and BiTeI

    KAUST Repository

    Zhu, Zhiyong; Cheng, Yingchun; Schwingenschlö gl, Udo

    2013-01-01

    By all-electron ab initio calculations, the layered polar semiconductor BiTeCl is shown to host giant bulk Rashba spin splitting, similar to the recently reported compound BiTeI. In both materials, the standard Rashba–Bychkov model is no longer applicable, because of huge band extrema shifts even in the absence of spin–orbit coupling and a strong momentum dependence of the Rashba coupling constant (αR). By assuming αR to be orbital dependent, a phenomenological extension of the Rashba–Bychkov model is proposed which explains the splitting behavior of states with small in-plane momentum.

  16. Gamma-ray yield dependence on bulk density and moisture content of a sample of a PGNAA setup. A Monte Carlo study

    International Nuclear Information System (INIS)

    Nagadi, M.M.; Naqvi, A.A.

    2007-01-01

    Monte Carlo calculations were carried out to study the dependence of γ-ray yield on the bulk density and moisture content of a sample in a thermalneutron capture-based prompt gamma neutron activation analysis (PGNAA) setup. The results of the study showed a strong dependence of the γ-ray yield upon the sample bulk density. An order of magnitude increase in yield of 1.94 and 6.42 MeV prompt γ-rays from calcium in a Portland cement sample was observed for a corresponding order of magnitude increase in the sample bulk density. On the contrary the γ-ray yield has a weak dependence on sample moisture content and an increase of only 20% in yield of 1.94 and 6.42 MeV prompt γ-rays from calcium in the Portland cement sample was observed for an order of magnitude increase in the moisture content of the Portland cement sample. A similar effect of moisture content has been observed on the yield of 1.167 MeV prompt γ-rays from chlorine contaminants in Portland cement samples. For an order of magnitude increase in the moisture content of the sample, a 7 to 12% increase in the yield of the 1.167 MeV chlorine γ-ray was observed for the Portland cement samples containing 1 to 5 wt.% chlorine contaminants. This study has shown that effects of sample moisture content on prompt γ-ray yield from constituents of a Portland cement sample are insignificant in a thermal-neutrons capture-based PGNAA setup. (author)

  17. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1.

    Science.gov (United States)

    Svensson, Katrin J; Christianson, Helena C; Wittrup, Anders; Bourseau-Guilmain, Erika; Lindqvist, Eva; Svensson, Lena M; Mörgelin, Matthias; Belting, Mattias

    2013-06-14

    The role of exosomes in cancer can be inferred from the observation that they transfer tumor cell derived genetic material and signaling proteins, resulting in e.g. increased tumor angiogenesis and metastasis. However, the membrane transport mechanisms and the signaling events involved in the uptake of these virus-like particles remain ill-defined. We now report that internalization of exosomes derived from glioblastoma (GBM) cells involves nonclassical, lipid raft-dependent endocytosis. Importantly, we show that the lipid raft-associated protein caveolin-1 (CAV1), in analogy with its previously described role in virus uptake, negatively regulates the uptake of exosomes. We find that exosomes induce the phosphorylation of several downstream targets known to associate with lipid rafts as signaling and sorting platforms, such as extracellular signal-regulated kinase-1/2 (ERK1/2) and heat shock protein 27 (HSP27). Interestingly, exosome uptake appears dependent on unperturbed ERK1/2-HSP27 signaling, and ERK1/2 phosphorylation is under negative influence by CAV1 during internalization of exosomes. These findings significantly advance our general understanding of exosome-mediated uptake and offer potential strategies for how this pathway may be targeted through modulation of CAV1 expression and ERK1/2 signaling.

  18. Modulation of iridovirus-induced apoptosis by endocytosis, early expression, JNK, and apical caspase

    International Nuclear Information System (INIS)

    Chitnis, Nilesh S.; D'Costa, Susan M.; Paul, Eric R.; Bilimoria, Shaen L.

    2008-01-01

    Chilo iridescent virus (CIV) is the type species for the family Iridoviridae, which are large, isometric, cytoplasmic dsDNA viruses. We examined the mechanism of apoptosis induction by CIV. High CIV doses (CIV XS ; 400 μg/ml), UV-irradiated virus (CIV UV ; 10 μg/ml) and CVPE (CIV protein extract; 10 μg/ml) induced apoptosis in 60% of treated Choristoneura fumiferana (IPRI-CF-124T) cells. Normal doses of infectious CIV (10 μg/ml) induced apoptosis in only 10% of C. fumiferana (CF) cells. Apoptosis was inhibited by Z-IETD-FMK, an apical caspase inhibitor, indicating that CIV-induced apoptosis requires caspase activity. The putative caspase in CF cells was designated Cf-caspase-i. CIV UV or CVPE enhanced Cf-caspase-i activity by 80% at 24 h relative to mock-treated cells. Since the MAP kinase pathway induces or inhibits apoptosis depending on the context, we used JNK inhibitor SP600125 and demonstrated drastic suppression of CVPE-induced apoptosis. Thus, the JNK signaling pathway is significant for apoptosis in this system. Virus interaction with the cell surface was not sufficient for apoptosis since CIV UV particles bound to polysterene beads failed to induce apoptosis. Endocytosis inhibitors (bafilomycin or ammonium chloride) negated apoptosis induction by CIV UV , CIV XS or CVPE indicating that entry through this mode is required. Given the weak apoptotic response to infectious CIV, we postulated that viral gene expression inhibited apoptosis. CIV infection of cells pretreated with cycloheximide induced apoptosis in 69% of the cells compared to 10% in normal infections. Furthermore, blocking viral DNA replication with aphidicolin or phosphonoacetic acid suppressed apoptosis and Cf-caspase-i activity, indicating that early viral expression is necessary for inhibition of apoptosis, and de novo synthesis of viral proteins is not required for induction. We show for the first time that, in a member of the family Iridoviridae, apoptosis: (i) requires entry and

  19. Effect of sulfur dioxide on pulmonary macrophage endocytosis at rest and during exercise

    International Nuclear Information System (INIS)

    Skornik, W.A.; Brain, J.D.

    1990-01-01

    Inhaled SO2 may cause damage by injuring upper airways. To what extent can SO2 also alter pulmonary macrophage function in the parenchyma and what is the impact of exercise? We studied the effect of SO2 on pulmonary macrophage endocytosis in resting and in exercising animals by measuring the rates of macrophage endocytosis in situ for 1 h of a test particle of insoluble radioactive colloidal gold (198Au), 1, 24, or 48 h after inhalation exposure to SO2. Resting hamsters exposed for 4 h to 50 ppm SO2 had no significant reduction in macrophage endocytosis compared with air-breathing control hamsters. However, if hamsters were exposed to the same concentration of SO2 while continuously running (40 min at 0.9 km/h), macrophage endocytosis was significantly reduced 1 h after exposure even though the exposure time was only one-sixth as long. Twenty-four hours later, the percentage of gold ingested by pulmonary macrophages remained significantly depressed. By 48 h, the rate had returned to control values. Exercise alone did not affect endocytosis. Hamsters exposed to 50 ppm SO2, with or without exercise, also showed significant reductions in the number of lavaged macrophages. This decrease was greatest and most persistent in the SO2 plus exercise group. These data indicate that even when animals are exposed to water-soluble gases, which are normally removed by the upper airways, exercise can potentiate damage to more peripheral components of the pulmonary defense system such as the macrophage

  20. Autophagy Proteins in Phagocyte Endocytosis and Exocytosis

    Directory of Open Access Journals (Sweden)

    Christian Münz

    2017-09-01

    Full Text Available Autophagy was initially described as a catabolic pathway that recycles nutrients of cytoplasmic constituents after lysosomal degradation during starvation. Since the immune system monitors products of lysosomal degradation via major histocompatibility complex (MHC class II restricted antigen presentation, autophagy was found to process intracellular antigens for display on MHC class II molecules. In recent years, however, it has become apparent that the molecular machinery of autophagy serves phagocytes in many more membrane trafficking pathways, thereby regulating immunity to infectious disease agents. In this minireview, we will summarize the recent evidence that autophagy proteins regulate phagocyte endocytosis and exocytosis for myeloid cell activation, pathogen replication, and MHC class I and II restricted antigen presentation. Selective stimulation and inhibition of the respective functional modules of the autophagy machinery might constitute valid therapeutic options in the discussed disease settings.

  1. TLR2 ligands induce NF-κB activation from endosomal compartments of human monocytes.

    Directory of Open Access Journals (Sweden)

    Karim J Brandt

    Full Text Available Localization of Toll-like receptors (TLR in subcellular organelles is a major strategy to regulate innate immune responses. While TLR4, a cell-surface receptor, signals from both the plasma membrane and endosomal compartments, less is known about the functional role of endosomal trafficking upon TLR2 signaling. Here we show that the bacterial TLR2 ligands Pam3CSK4 and LTA activate NF-κB-dependent signaling from endosomal compartments in human monocytes and in a NF-κB sensitive reporter cell line, despite the expression of TLR2 at the cell surface. Further analyses indicate that TLR2-induced NF-κB activation is controlled by a clathrin/dynamin-dependent endocytosis mechanism, in which CD14 serves as an important upstream regulator. These findings establish that internalization of cell-surface TLR2 into endosomal compartments is required for NF-κB activation. These observations further demonstrate the need of endocytosis in the activation and regulation of TLR2-dependent signaling pathways.

  2. Temperature and driving field dependence of fatigue processes in PZT bulk ceramics

    International Nuclear Information System (INIS)

    Glaum, Julia; Granzow, Torsten; Schmitt, Ljubomira Ana; Kleebe, Hans-Joachim; Roedel, Juergen

    2011-01-01

    The temperature- and field-dependent degradation properties of bulk Pb(Zr,Ti)O 3 material (PZT) under a unipolar electric field were investigated. Unipolar cycling leads to the build-up of an internal bias field based on the agglomeration of charges at grain boundaries. A simple model was developed which describes the general dynamics of unipolar fatigue and its dependence on temperature and driving field. Comparing the large and small signal permittivity before and after fatigue led to the conclusion that domain walls became clamped by the agglomerated charges. This clamping effect could be visualized by transmission electron microscopy (TEM). Additionally, the TEM investigations revealed that unipolar fatigue leads to a weakening of the microstructure and to the development of microcracks.

  3. Kinetics of cellular uptake of viruses and nanoparticles via clathrin-mediated endocytosis

    Science.gov (United States)

    Banerjee, Anand; Berezhkovskii, Alexander; Nossal, Ralph

    2016-02-01

    Several viruses exploit clathrin-mediated endocytosis to gain entry into host cells. This process is also used extensively in biomedical applications to deliver nanoparticles (NPs) to diseased cells. The internalization of these nano-objects is controlled by the assembly of a clathrin-containing protein coat on the cytoplasmic side of the plasma membrane, which drives the invagination of the membrane and the formation of a cargo-containing endocytic vesicle. Current theoretical models of receptor-mediated endocytosis of viruses and NPs do not explicitly take coat assembly into consideration. In this paper we study cellular uptake of viruses and NPs with a focus on coat assembly. We characterize the internalization process by the mean time between the binding of a particle to the membrane and its entry into the cell. Using a coarse-grained model which maps the stochastic dynamics of coat formation onto a one-dimensional random walk, we derive an analytical formula for this quantity. A study of the dependence of the mean internalization time on NP size shows that there is an upper bound above which this time becomes extremely large, and an optimal size at which it attains a minimum. Our estimates of these sizes compare well with experimental data. We also study the sensitivity of the obtained results on coat parameters to identify factors which significantly affect the internalization kinetics.

  4. Effect of sample moisture and bulk density on performance of the 241Am-Be source based prompt gamma rays neutron activation analysis setup. A Monte Carlo study

    International Nuclear Information System (INIS)

    Almisned, Ghada

    2010-01-01

    Monte Carlo simulations were carried out using the dependence of gamma ray yield on the bulk density and moisture content for five different lengths of Portland cement samples in a thermal neutron capture based Prompt Gamma ray Neutron Activation Analysis (PGNAA) setup for source inside moderator geometry using an 241 Am-Be neutron source. In this study, yields of 1.94 and 6.42 MeV prompt gamma rays from calcium in the five Portland cement samples were calculated as a function of sample bulk density and moisture content. The study showed a strong dependence of the 1.94 and 6.42 MeV gamma ray yield upon the sample bulk density but a weaker dependence upon sample moisture content. For an order of magnitude increase in the sample bulk density, an order of magnitude increase in the gamma rays yield was observed, i.e., a one-to-one correspondence. In case of gamma ray yield dependence upon sample moisture content, an order of magnitude increase in the moisture content of the sample resulted in about 16-17% increase in the yield of 1.94 and 6.42 MeV gamma rays from calcium. (author)

  5. Meeting after meeting: 20 years of discoveries by the members of the Exocytosis-Endocytosis Club.

    Science.gov (United States)

    Niedergang, Florence; Gasman, Stéphane; Vitale, Nicolas; Desnos, Claire; Lamaze, Christophe

    2017-09-01

    Twenty years ago, a group of French cell biologists merged two scientific clubs with the aim of bringing together researchers in the fields of Endocytosis and Exocytosis. Founded in 1997, the first annual meeting of the Exocytosis Club was held in 1998. The Endocytosis Club held quarterly meetings from its founding in 1999. The first joint annual meeting of the Exocytosis-Endocytosis Club took place in Paris in April, 2001. What started as a modest gathering of enthusiastic scientists working in the field of cell trafficking has gone from strength to strength, rapidly becoming an unmissable yearly meeting, vividly demonstrating the high quality of science performed in our community and beyond. On the occasion of the 20th meeting of our club, we want to provide historic insight into the fields of exocytosis and endocytosis, and by extension, to subcellular trafficking, highlighting how French scientists have contributed to major advances in these fields. Today, the Exocytosis-Endocytosis Club represents a vibrant and friendly community that will hold its 20th meeting at the Presqu'Ile de Giens, near Toulon in the South of France, on May 11-13, 2017. © 2017 Société Française des Microscopies and Société de Biologie Cellulaire de France. Published by John Wiley & Sons Ltd.

  6. The Lowe syndrome protein OCRL1 is required for endocytosis in the zebrafish pronephric tubule.

    Directory of Open Access Journals (Sweden)

    Francesca Oltrabella

    2015-04-01

    Full Text Available Lowe syndrome and Dent-2 disease are caused by mutation of the inositol 5-phosphatase OCRL1. Despite our increased understanding of the cellular functions of OCRL1, the underlying basis for the renal tubulopathy seen in both human disorders, of which a hallmark is low molecular weight proteinuria, is currently unknown. Here, we show that deficiency in OCRL1 causes a defect in endocytosis in the zebrafish pronephric tubule, a model for the mammalian renal tubule. This coincides with a reduction in levels of the scavenger receptor megalin and its accumulation in endocytic compartments, consistent with reduced recycling within the endocytic pathway. We also observe reduced numbers of early endocytic compartments and enlarged vacuolar endosomes in the sub-apical region of pronephric cells. Cell polarity within the pronephric tubule is unaffected in mutant embryos. The OCRL1-deficient embryos exhibit a mild ciliogenesis defect, but this cannot account for the observed impairment of endocytosis. Catalytic activity of OCRL1 is required for renal tubular endocytosis and the endocytic defect can be rescued by suppression of PIP5K. These results indicate for the first time that OCRL1 is required for endocytic trafficking in vivo, and strongly support the hypothesis that endocytic defects are responsible for the renal tubulopathy in Lowe syndrome and Dent-2 disease. Moreover, our results reveal PIP5K as a potential therapeutic target for Lowe syndrome and Dent-2 disease.

  7. AC conductivity and dielectric behavior of bulk Furfurylidenemalononitrile

    Science.gov (United States)

    El-Nahass, M. M.; Ali, H. A. M.

    2012-06-01

    AC conductivity and dielectric behavior for bulk Furfurylidenemalononitrile have been studied over a temperature range (293-333 K) and frequency range (50-5×106 Hz). The frequency dependence of ac conductivity, σac, has been investigated by the universal power law, σac(ω)=Aωs. The variation of the frequency exponent (s) with temperature was analyzed in terms of different conduction mechanisms, and it was found that the correlated barrier hopping (CBH) model is the predominant conduction mechanism. The temperature dependence of σac(ω) showed a linear increase with the increase in temperature at different frequencies. The ac activation energy was determined at different frequencies. Dielectric data were analyzed using complex permittivity and complex electric modulus for bulk Furfurylidenemalononitrile at various temperatures.

  8. Avr4 promotes Cf-4 receptor-like protein association with the BAK1/SERK3 receptor-like kinase to initiate receptor endocytosis and plant immunity.

    Science.gov (United States)

    Postma, Jelle; Liebrand, Thomas W H; Bi, Guozhi; Evrard, Alexandre; Bye, Ruby R; Mbengue, Malick; Kuhn, Hannah; Joosten, Matthieu H A J; Robatzek, Silke

    2016-04-01

    The first layer of plant immunity is activated by cell surface receptor-like kinases (RLKs) and proteins (RLPs) that detect infectious pathogens. Constitutive interaction with the SUPPRESSOR OF BIR1 (SOBIR1) RLK contributes to RLP stability and kinase activity. As RLK activation requires transphosphorylation with a second associated RLK, it remains elusive how RLPs initiate downstream signaling. We employed live-cell imaging, gene silencing and coimmunoprecipitation to investigate the requirement of associated kinases for functioning and ligand-induced subcellular trafficking of Cf RLPs that mediate immunity of tomato against Cladosporium fulvum. Our research shows that after elicitation with matching effector ligands Avr4 and Avr9, BRI1-ASSOCIATED KINASE 1/SOMATIC EMBRYOGENESIS RECEPTOR KINASE 3 (BAK1/SERK3) associates with Cf-4 and Cf-9. BAK1/SERK3 is required for the effector-triggered hypersensitive response and resistance of tomato against C. fulvum. Furthermore, Cf-4 interacts with SOBIR1 at the plasma membrane and is recruited to late endosomes upon Avr4 trigger, also depending on BAK1/SERK3. These observations indicate that RLP-mediated resistance and endocytosis require ligand-induced recruitment of BAK1/SERK3, reminiscent of BAK1/SERK3 interaction and subcellular fate of the FLAGELLIN SENSING 2 (FLS2) RLK. This reveals that diverse classes of cell surface immune receptors share common requirements for initiation of resistance and endocytosis. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  9. Dimerization and endocytosis of the sucrose transporter StSUT1 in mature sieve elements.

    Science.gov (United States)

    Liesche, Johannes; Schulz, Alexander; Krügel, Undine; Grimm, Bernhard; Kühn, Christina

    2008-12-01

    The sucrose transporter StSUT1 from Solanum tuberosum was shown to be regulated post-translationally by redox reagents. Its activity is increased at least 10-fold in the presence of oxidizing agents if expressed in yeast. Oxidation has also an effect on plasma membrane targeting and dimerization of the protein. In response to oxidizing agents, StSUT1 is targeted to lipid raft-like microdomains and SUT1 protein is detectable in the detergent resistant membrane fraction of plant plasma membranes. Interestingly, StSUT1 treated with brefeldin A seems to aggregate in endocytic compartments in mature sieve elements.1 Further analysis of SUT1 targeting will certainly provide more information about the putative involvement of lipid raft-like microdomains in endocytic events. We provide here additional information on the dimerization and endocytosis of the SUT1 protein. The oligomerization of overexpressed SoSUT1 from Spinacia oleracea in transgenic potato plants was analyzed by two-dimensional gel electrophoresis and endocytosis of the StSUT1 protein was confirmed by immunogold labeling.

  10. Simultaneous measurement of thermal conductivity and heat capacity of bulk and thin film materials using frequency-dependent transient thermoreflectance method.

    Science.gov (United States)

    Liu, Jun; Zhu, Jie; Tian, Miao; Gu, Xiaokun; Schmidt, Aaron; Yang, Ronggui

    2013-03-01

    The increasing interest in the extraordinary thermal properties of nanostructures has led to the development of various measurement techniques. Transient thermoreflectance method has emerged as a reliable measurement technique for thermal conductivity of thin films. In this method, the determination of thermal conductivity usually relies much on the accuracy of heat capacity input. For new nanoscale materials with unknown or less-understood thermal properties, it is either questionable to assume bulk heat capacity for nanostructures or difficult to obtain the bulk form of those materials for a conventional heat capacity measurement. In this paper, we describe a technique for simultaneous measurement of thermal conductivity κ and volumetric heat capacity C of both bulk and thin film materials using frequency-dependent time-domain thermoreflectance (TDTR) signals. The heat transfer model is analyzed first to find how different combinations of κ and C determine the frequency-dependent TDTR signals. Simultaneous measurement of thermal conductivity and volumetric heat capacity is then demonstrated with bulk Si and thin film SiO2 samples using frequency-dependent TDTR measurement. This method is further testified by measuring both thermal conductivity and volumetric heat capacity of novel hybrid organic-inorganic thin films fabricated using the atomic∕molecular layer deposition. Simultaneous measurement of thermal conductivity and heat capacity can significantly shorten the development∕discovery cycle of novel materials.

  11. Microparticle-Induced Activation of the Vascular Endothelium Requires Caveolin-1/Caveolae.

    Directory of Open Access Journals (Sweden)

    Allison M Andrews

    Full Text Available Microparticles (MPs are small membrane fragments shed from normal as well as activated, apoptotic or injured cells. Emerging evidence implicates MPs as a causal and/or contributing factor in altering normal vascular cell phenotype through initiation of proinflammatory signal transduction events and paracrine delivery of proteins, mRNA and miRNA. However, little is known regarding the mechanism by which MPs influence these events. Caveolae are important membrane microdomains that function as centers of signal transduction and endocytosis. Here, we tested the concept that the MP-induced pro-inflammatory phenotype shift in endothelial cells (ECs depends on caveolae. Consistent with previous reports, MP challenge activated ECs as evidenced by upregulation of intracellular adhesion molecule-1 (ICAM-1 expression. ICAM-1 upregulation was mediated by activation of NF-κB, Poly [ADP-ribose] polymerase 1 (PARP-1 and the epidermal growth factor receptor (EGFR. This response was absent in ECs lacking caveolin-1/caveolae. To test whether caveolae-mediated endocytosis, a dynamin-2 dependent process, is a feature of the proinflammatory response, EC's were pretreated with the dynamin-2 inhibitor dynasore. Similar to observations in cells lacking caveolin-1, inhibition of endocytosis significantly attenuated MPs effects including, EGFR phosphorylation, activation of NF-κB and upregulation of ICAM-1 expression. Thus, our results indicate that caveolae play a role in mediating the pro-inflammatory signaling pathways which lead to EC activation in response to MPs.

  12. EH and UIM: endocytosis and more

    DEFF Research Database (Denmark)

    Polo, Simona; Confalonieri, Stefano; Salcini, Anna Elisabetta

    2003-01-01

    Exogenously and endogenously originated signals are propagated within the cell by functional and physical networks of proteins, leading to numerous biological outcomes. Many protein-protein interactions take place between binding domains and short peptide motifs. Frequently, these interactions......). The other, which we define as the monoubiquitin (mUb) network, relies on monoubiquitination, which is emerging as an important posttranslational modification that regulates protein function. Both networks were initially implicated in the control of plasma membrane receptor endocytosis and in the regulation...

  13. Flavivirus internalization is regulated by a size-dependent endocytic pathway.

    Science.gov (United States)

    Hackett, Brent A; Cherry, Sara

    2018-04-17

    Flaviviruses enter host cells through the process of clathrin-mediated endocytosis, and the spectrum of host factors required for this process are incompletely understood. Here we found that lymphocyte antigen 6 locus E (LY6E) promotes the internalization of multiple flaviviruses, including West Nile virus, Zika virus, and dengue virus. Perhaps surprisingly, LY6E is dispensable for the internalization of the endogenous cargo transferrin, which is also dependent on clathrin-mediated endocytosis for uptake. Since viruses are substantially larger than transferrin, we reasoned that LY6E may be required for uptake of larger cargoes and tested this using transferrin-coated beads of similar size as flaviviruses. LY6E was indeed required for the internalization of transferrin-coated beads, suggesting that LY6E is selectively required for large cargo. Cell biological studies found that LY6E forms tubules upon viral infection and bead internalization, and we found that tubule formation was dependent on RNASEK, which is also required for flavivirus internalization, but not transferrin uptake. Indeed, we found that RNASEK is also required for the internalization of transferrin-coated beads, suggesting it functions upstream of LY6E. These LY6E tubules resembled microtubules, and we found that microtubule assembly was required for their formation and flavivirus uptake. Since microtubule end-binding proteins link microtubules to downstream activities, we screened the three end-binding proteins and found that EB3 promotes virus uptake and LY6E tubularization. Taken together, these results highlight a specialized pathway required for the uptake of large clathrin-dependent endocytosis cargoes, including flaviviruses. Copyright © 2018 the Author(s). Published by PNAS.

  14. Duffy antigen receptor for chemokines mediates chemokine endocytosis through a macropinocytosis-like process in endothelial cells.

    Directory of Open Access Journals (Sweden)

    Yani Zhao

    Full Text Available The Duffy antigen receptor for chemokines (DARC shows high affinity binding to multiple inflammatory CC and CXC chemokines and is expressed by erythrocytes and endothelial cells. Recent evidence suggests that endothelial DARC facilitates chemokine transcytosis to promote neutrophil recruitment. However, the mechanism of chemokine endocytosis by DARC remains unclear.We investigated the role of several endocytic pathways in DARC-mediated ligand internalization. Here we report that, although DARC co-localizes with caveolin-1 in endothelial cells, caveolin-1 is dispensable for DARC-mediated (125I-CXCL1 endocytosis as knockdown of caveolin-1 failed to inhibit ligand internalization. (125I-CXCL1 endocytosis by DARC was also independent of clathrin and flotillin-1 but required cholesterol and was, in part, inhibited by silencing Dynamin II expression.(125I-CXCL1 endocytosis was inhibited by amiloride, cytochalasin D, and the PKC inhibitor Gö6976 whereas Platelet Derived Growth Factor (PDGF enhanced ligand internalization through DARC. The majority of DARC-ligand interactions occurred on the endothelial surface, with DARC identified along plasma membrane extensions with the appearance of ruffles, supporting the concept that DARC provides a high affinity scaffolding function for surface retention of chemokines on endothelial cells.These results show DARC-mediated chemokine endocytosis occurs through a macropinocytosis-like process in endothelial cells and caveolin-1 is dispensable for CXCL1 internalization.

  15. Improved α-Amylase Production by Dephosphorylation Mutation of CreD, an Arrestin-Like Protein Required for Glucose-Induced Endocytosis of Maltose Permease and Carbon Catabolite Derepression in Aspergillus oryzae.

    Science.gov (United States)

    Tanaka, Mizuki; Hiramoto, Tetsuya; Tada, Hinako; Shintani, Takahiro; Gomi, Katsuya

    2017-07-01

    Aspergillus oryzae produces copious amount of amylolytic enzymes, and MalP, a major maltose permease, is required for the expression of amylase-encoding genes. The expression of these genes is strongly repressed by carbon catabolite repression (CCR) in the presence of glucose. MalP is transported from the plasma membrane to the vacuole by endocytosis, which requires the homolog of E6-AP carboxyl terminus ubiquitin ligase HulA, an ortholog of yeast Rsp5. In yeast, arrestin-like proteins mediate endocytosis as adaptors of Rsp5 and transporters. In the present study, we examined the involvement of CreD, an arrestin-like protein, in glucose-induced MalP endocytosis and CCR of amylase-encoding genes. Deletion of creD inhibited the glucose-induced endocytosis of MalP, and CreD showed physical interaction with HulA. Phosphorylation of CreD was detected by Western blotting, and two serine residues were determined as the putative phosphorylation sites. However, the phosphorylation state of the serine residues did not regulate MalP endocytosis and its interaction with HulA. Although α-amylase production was significantly repressed by creD deletion, both phosphorylation and dephosphorylation mimics of CreD had a negligible effect on α-amylase activity. Interestingly, dephosphorylation of CreD was required for CCR relief of amylase genes that was triggered by disruption of the deubiquitinating enzyme-encoding gene creB The α-amylase activity of the creB mutant was 1.6-fold higher than that of the wild type, and the dephosphorylation mimic of CreD further improved the α-amylase activity by 2.6-fold. These results indicate that a combination of the dephosphorylation mutation of CreD and creB disruption increased the production of amylolytic enzymes in A. oryzae IMPORTANCE In eukaryotes, glucose induces carbon catabolite repression (CCR) and proteolytic degradation of plasma membrane transporters via endocytosis. Glucose-induced endocytosis of transporters is mediated by

  16. Synaptophysin 1 Clears Synaptobrevin 2 from the Presynaptic Active Zone to Prevent Short-Term Depression

    Directory of Open Access Journals (Sweden)

    Rajit Rajappa

    2016-02-01

    Full Text Available Release site clearance is an important process during synaptic vesicle (SV recycling. However, little is known about its molecular mechanism. Here we identify self-assembly of exocytosed Synaptobrevin 2 (Syb2 and Synaptophysin 1 (Syp1 by homo- and hetero-oligomerization into clusters as key mechanisms mediating release site clearance for preventing cis-SNARE complex formation at the active zone (AZ. In hippocampal neurons from Syp1 knockout mice, neurons expressing a monomeric Syb2 mutant, or after acute block of the ATPase N-ethylmaleimide-sensitive factor (NSF, responsible for cis-SNARE complex disassembly, we found strong frequency-dependent short-term depression (STD, whereas retrieval of Syb2 by compensatory endocytosis was only affected weakly. Defects in Syb2 endocytosis were stimulus- and frequency-dependent, indicating that Syp1 is not essential for Syb2 retrieval, but for its efficient clearance upstream of endocytosis. Our findings identify an SV protein as a release site clearance factor.

  17. Management experience on microthrix parvicella bulking in an activated sludge wastewater treatment plant

    International Nuclear Information System (INIS)

    De Bortoli, N.; Mion, M.; Di Giorgio, G.; Goi, D.

    2005-01-01

    Activated sludge wastewater treatment processes may give inefficiencies due to biological imbalances involving biomass. In fact, external causes as temperature lowering can increase the proliferation of the filamentous bacterium Microthrix parvicella into activated sludge flocks. Microthrix parvicella increases may create dangerous bulking phenomena compromising secondary settling without varying bio-kinetic parameters. In this case of study, a method to defeat growth of Microthrix parvicella has been set up. Aluminium poly-chloride (PAC) has been added to activated sludge contained into oxidation tanks of a municipal wastewater treatment plant, where a large growth of Microthrix parvicella has been periodically observed. It has been demonstrated that a definite PAC concentration can reduce Microthrix parvicella proliferation into activated sludge flocks so bulking phenomena can be well reduced [it

  18. Zebrafish kidney phagocytes utilize macropinocytosis and Ca+-dependent endocytic mechanisms.

    Directory of Open Access Journals (Sweden)

    Claudia Hohn

    Full Text Available BACKGROUND: The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including active endocytosis by macrophages and granulocytes. Endocytosis can be used as a reliable measure of selective and non-selective mechanisms of antigen uptake in the early phase of an immune response. Numerous assays have been developed to measure this response in a variety of mammalian and fish species. The small size of the zebrafish has prevented the large-scale collection of monocytes/macrophages and granulocytes for these endocytic assays. METHODOLOGY/PRINCIPAL FINDINGS: Pooled zebrafish kidney hematopoietic tissues were used as a source of phagocytic cells for flow-cytometry based endocytic assays. FITC-Dextran, Lucifer Yellow and FITC-Edwardsiella ictaluri were used to evaluate selective and non-selective mechanisms of uptake in zebrafish phagocytes. CONCLUSIONS/SIGNIFICANCE: Zebrafish kidney phagocytes characterized as monocytes/macrophages, neutrophils and lymphocytes utilize macropinocytosis and Ca(2+-dependant endocytosis mechanisms of antigen uptake. These cells do not appear to utilize a mannose receptor. Heat-killed Edwardsiella ictaluri induces cytoskeletal interactions for internalization in zebrafish kidney monocytes/macrophages and granulocytes. The proposed method is easy to implement and should prove especially useful in immunological, toxicological and epidemiological research.

  19. Temperature dependence of the mechanical properties of melt-processed Dy-Ba-Cu-O bulk superconductors evaluated by three point bending tests

    International Nuclear Information System (INIS)

    Katagiri, K; Nyilas, A; Sato, T; Hatakeyama, Y; Hokari, T; Teshima, H; Iwamoto, A; Mito, T

    2006-01-01

    Dy-Ba-Cu-O bulk superconductor has an excellent capability of trapping magnetic flux and lower heat conductivity at cryogenic temperatures as compared with Y-Ba-Cu-O bulk superconductor. The Young's modulus and the bending strength in the range from room temperature to 7 K were measured by the three-point bending tests using specimens cut from a melt-processed Dy-Ba-Cu-O bulk superconductor. They were tested in a helium gas flow type cryostat at Forschungszentrum Karlsruhe and in a liquid nitrogen bath at Iwate University. The Young's modulus was calculated by either the slope of stress-strain curve or that of the load-deflection curve of the specimen. Although the bending strength measured in the two institutes coincided well, there was a significant discrepancy in the Young's modulus. The Young's modulus and bending strength increased with decrease of temperature down to 7 K. The amount of increase in the Young's modulus and the bending strength were about 32% and 36% of those at room temperature, respectively. The scatter of data for each run was significant and did not depend on temperature. The temperature dependence of the Young's modulus coincided with that in Y-Ba-Cu-O obtained by ultrasonic velocity. The temperature dependence of the Young's modulus and the bending strength was discussed from the view point of interatomic distance of the bulk crystal

  20. Quantitative proteome analysis reveals the correlation between endocytosis-associated proteins and hepatocellular carcinoma dedifferentiation.

    Science.gov (United States)

    Naboulsi, Wael; Bracht, Thilo; Megger, Dominik A; Reis, Henning; Ahrens, Maike; Turewicz, Michael; Eisenacher, Martin; Tautges, Stephanie; Canbay, Ali E; Meyer, Helmut E; Weber, Frank; Baba, Hideo A; Sitek, Barbara

    2016-11-01

    The majority of poorly differentiated hepatocellular carcinomas (HCCs) develop from well-differentiated tumors. Endocytosis is a cellular function which is likely to take part in this development due to its important role in regulating the abundances of vital signaling receptors. Here, we aimed to investigate the abundance of endocytosis-associated proteins in HCCs with various differentiation grades. Therefore, we analyzed 36 tissue specimens from HCC patients via LC-MS/MS-based label-free quantitative proteomics including 19 HCC tissue samples with different degrees of histological grades and corresponding non-tumorous tissue controls. As a result, 277 proteins were differentially regulated between well-differentiated tumors and controls. In moderately and poorly differentiated tumors, 278 and 1181 proteins, respectively, were significantly differentially regulated compared to non-tumorous tissue. We explored the regulated proteins based on their functions and identified thirty endocytosis-associated proteins, mostly overexpressed in poorly differentiated tumors. These included proteins that have been shown to be up-regulated in HCC like clathrin heavy chain-1 (CLTC) as well as unknown proteins, such as secretory carrier-associated membrane protein 3 (SCAMP3). The abundances of SCAMP3 and CLTC were immunohistochemically examined in tissue sections of 84 HCC patients. We demonstrate the novel association of several endocytosis-associated proteins, in particular, SCAMP3 with HCC progression. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Clathrin-independent endocytosis: mechanisms and function

    DEFF Research Database (Denmark)

    Sandvig, Kirsten; Pust, Sascha; Skotland, Tore

    2011-01-01

    It is now about 20 years since we first wrote reviews about clathrin-independent endocytosis. The challenge at the time was to convince the reader about its existence. Then the suggestion came up that caveolae might be responsible for the uptake. However, clearly this could not be the case since ...... having several functions of their own. This article aims at providing a brief update on the importance of clathrin-independent endocytic mechanisms, how the processes are regulated differentially, for instance on the poles of polarized cells, and the challenges in studying them....

  2. The Measles Virus Receptor SLAMF1 Can Mediate Particle Endocytosis.

    Science.gov (United States)

    Gonçalves-Carneiro, Daniel; McKeating, Jane A; Bailey, Dalan

    2017-04-01

    The signaling lymphocyte activation molecule F1 (SLAMF1) is both a microbial sensor and entry receptor for measles virus (MeV). Herein, we describe a new role for SLAMF1 to mediate MeV endocytosis that is in contrast with the alternative, and generally accepted, model that MeV genome enters cells only after fusion at the cell surface. We demonstrated that MeV engagement of SLAMF1 induces dramatic but transient morphological changes, most prominently in the formation of membrane blebs, which were shown to colocalize with incoming viral particles, and rearrangement of the actin cytoskeleton in infected cells. MeV infection was dependent on these dynamic cytoskeletal changes as well as fluid uptake through a macropinocytosis-like pathway as chemical inhibition of these processes inhibited entry. Moreover, we identified a role for the RhoA-ROCK-myosin II signaling axis in this MeV internalization process, highlighting a novel role for this recently characterized pathway in virus entry. Our study shows that MeV can hijack a microbial sensor normally involved in bacterial phagocytosis to drive endocytosis using a complex pathway that shares features with canonical viral macropinocytosis, phagocytosis, and mechanotransduction. This uptake pathway is specific to SLAMF1-positive cells and occurs within 60 min of viral attachment. Measles virus remains a significant cause of mortality in human populations, and this research sheds new light on the very first steps of infection of this important pathogen. IMPORTANCE Measles is a significant disease in humans and is estimated to have killed over 200 million people since records began. According to current World Health Organization statistics, it still kills over 100,000 people a year, mostly children in the developing world. The causative agent, measles virus, is a small enveloped RNA virus that infects a broad range of cells during infection. In particular, immune cells are infected via interactions between glycoproteins found

  3. PGE2 mediates EGFR internalization and nuclear translocation via caveolin endocytosis promoting its transcriptional activity and proliferation in human NSCLC cells.

    Science.gov (United States)

    Bazzani, Lorenzo; Donnini, Sandra; Giachetti, Antonio; Christofori, Gerhard; Ziche, Marina

    2018-03-13

    Prostaglandin E 2 (PGE 2 ) contributes to tumor progression by promoting cancer cell growth, invasion and by creating a favorable pro-tumor microenvironment. PGE 2 has been reported to transactivate and internalize into the nucleus receptor tyrosine kinases such as Epidermal growth factor receptor (EGFR), thereby supporting tumor progression. Here we demonstrate that in non-small cell lung carcinoma (NSCLC) cells, PGE 2 induces EGFR nuclear translocation via different dynamin-dependent endocytic pathways, promotes the formation of an EGFR-STAT3 complex, affects nuclear EGFR target gene expression and mediates tumor cell proliferation. Indeed, we find that PGE 2 induces EGFR internalization and consequent nuclear import through Clathrin- and Caveolin-mediated endocytosis and through the interaction of EGFR with Importin β1. Within the nucleus, EGFR forms a complex with STAT3, an event blocked by ablation of Clathrin Heavy Chain or Caveolin-1. The combination of EGF and PGE 2 prolongs nuclear EGFR transcriptional activity manifested by the upregulation of CCND1 , PTGS2 , MYC and NOS2 mRNA levels and potentiates nuclear EGFR-induced NSCLC cell proliferation. Additionally, NSCLC patients with high expression of a nuclear EGFR gene signature display shorter survival times than those with low expression, thus showing a putative correlation between nuclear EGFR and poor prognosis in NSCLC. Together, our findings indicate a complex mechanism underlying PGE 2 -induced EGF/EGFR signaling and transcriptional control, which plays a key role in cancer progression.

  4. Shedding light on endocytosis with optimized super-resolution microscopy

    NARCIS (Netherlands)

    Leyton Puig, D.M.

    2017-01-01

    Super-resolution microscopy is a relatively new microscopy technique that is still under optimization. In this thesis we focus on the improvement of the quality of super-resolution images, to apply them to the study of the processes of cell signaling and endocytosis. First, we show that the use of a

  5. Analysis of occludin trafficking, demonstrating continuous endocytosis, degradation, recycling and biosynthetic secretory trafficking.

    Directory of Open Access Journals (Sweden)

    Sarah J Fletcher

    Full Text Available Tight junctions (TJs link adjacent cells and are critical for maintenance of apical-basolateral polarity in epithelial monolayers. The TJ protein occludin functions in disparate processes, including wound healing and Hepatitis C Virus infection. Little is known about steady-state occludin trafficking into and out of the plasma membrane. Therefore, we determined the mechanisms responsible for occludin turnover in confluent Madin-Darby canine kidney (MDCK epithelial monolayers. Using various biotin-based trafficking assays we observed continuous and rapid endocytosis of plasma membrane localised occludin (the majority internalised within 30 minutes. By 120 minutes a significant reduction in internalised occludin was observed. Inhibition of lysosomal function attenuated the reduction in occludin signal post-endocytosis and promoted co-localisation with the late endocytic system. Using a similar method we demonstrated that ∼20% of internalised occludin was transported back to the cell surface. Consistent with these findings, significant co-localisation between internalised occludin and recycling endosomal compartments was observed. We then quantified the extent to which occludin synthesis and transport to the plasma membrane contributes to plasma membrane occludin homeostasis, identifying inhibition of protein synthesis led to decreased plasma membrane localised occludin. Significant co-localisation between occludin and the biosynthetic secretory pathway was demonstrated. Thus, under steady-state conditions occludin undergoes turnover via a continuous cycle of endocytosis, recycling and degradation, with degradation compensated for by biosynthetic exocytic trafficking. We developed a mathematical model to describe the endocytosis, recycling and degradation of occludin, utilising experimental data to provide quantitative estimates for the rates of these processes.

  6. Protein kinase A-induced internalization of Slack channels from the neuronal membrane occurs by adaptor protein-2/clathrin-mediated endocytosis.

    Science.gov (United States)

    Gururaj, Sushmitha; Evely, Katherine M; Pryce, Kerri D; Li, Jun; Qu, Jun; Bhattacharjee, Arin

    2017-11-24

    The sodium-activated potassium (K Na ) channel Kcnt1 (Slack) is abundantly expressed in nociceptor (pain-sensing) neurons of the dorsal root ganglion (DRG), where they transmit the large outward conductance I KNa and arbitrate membrane excitability. Slack channel expression at the DRG membrane is necessary for their characteristic firing accommodation during maintained stimulation, and reduced membrane channel density causes hyperexcitability. We have previously shown that in a pro-inflammatory state, a decrease in membrane channel expression leading to reduced Slack-mediated I KNa expression underlies DRG neuronal sensitization. An important component of the inflammatory milieu, PKA internalizes Slack channels from the DRG membrane, reduces I KNa , and produces DRG neuronal hyperexcitability when activated in cultured primary DRG neurons. Here, we show that this PKA-induced retrograde trafficking of Slack channels also occurs in intact spinal cord slices and that it is carried out by adaptor protein-2 (AP-2) via clathrin-mediated endocytosis. We provide mass spectrometric and biochemical evidence of an association of native neuronal AP-2 adaptor proteins with Slack channels, facilitated by a dileucine motif housed in the cytoplasmic Slack C terminus that binds AP-2. By creating a competitive peptide blocker of AP-2-Slack binding, we demonstrated that this interaction is essential for clathrin recruitment to the DRG membrane, Slack channel endocytosis, and DRG neuronal hyperexcitability after PKA activation. Together, these findings uncover AP-2 and clathrin as players in Slack channel regulation. Given the significant role of Slack in nociceptive neuronal excitability, the AP-2 clathrin-mediated endocytosis trafficking mechanism may enable targeting of peripheral and possibly, central neuronal sensitization. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Interaction of 125I-labeled botulinum neurotoxins with nerve terminals. II. Autoradiographic evidence for its uptake into motor nerves by acceptor-mediated endocytosis

    International Nuclear Information System (INIS)

    Black, J.D.; Dolly, J.O.

    1986-01-01

    Using pharmacological and autoradiographic techniques it has been shown that botulinum neurotoxin (BoNT) is translocated across the motor nerve terminal membrane to reach a postulated intraterminal target. In the present study, the nature of this uptake process was investigated using electron microscopic autoradiography. It was found that internalization is acceptor-mediated and that binding to specific cell surface acceptors involves the heavier chain of the toxin. In addition, uptake was shown to be energy and temperature-dependent and to be accelerated by nerve stimulation, a treatment which also shortens the time course of the toxin-induced neuroparalysis. These results, together with the observation that silver grains were often associated with endocytic structures within the nerve terminal, suggested that acceptor-mediated endocytosis is responsible for toxin uptake. Possible recycling of BoNT acceptors (an important aspect of acceptor-mediated endocytosis of toxins) at motor nerve terminals was indicated by comparing the extent of labeling in the presence and absence of metabolic inhibitors. On the basis of these collective results, it is concluded that BoNT is internalized by acceptor-mediated endocytosis and, hence, the data support the proposal that this toxin inhibits release of acetylcholine by interaction with an intracellular target

  8. Endophilin mutations block clathrin-mediated endocytosis but not neurotransmitter release

    DEFF Research Database (Denmark)

    Verstreken, Patrik; Kjaerulff, Ole; Lloyd, Thomas E

    2002-01-01

    We have identified mutations in Drosophila endophilin to study its function in vivo. Endophilin is required presynaptically at the neuromuscular junction, and absence of Endophilin dramatically impairs endocytosis in vivo. Mutant larvae that lack Endophilin fail to take up FM1-43 dye in synaptic ...

  9. Thickness dependence of the levitation performance of double-layer high-temperature superconductor bulks above a magnetic rail

    International Nuclear Information System (INIS)

    Sun, R.X.; Zheng, J.; Liao, X.L.; Che, T.; Gou, Y.F.; He, D.B.; Deng, Z.G.

    2014-01-01

    Highlights: • Thickness optimization of double-layer bulk HTSC arrangement is studied. • The new bulk HTSC arrangement makes better use of the flux distribution of the magnetic rails. • Levitation performance can be enhanced with the optimization. • The optimization can meet large levitation force requirements for HTS Maglev system. - Abstract: A double-layer high-temperature superconductor (HTSC) arrangement was proposed and proved to be able to bring improvements to both levitation force and guidance force compared with present single-layer HTSC arrangement. To fully exploit the applied magnetic field by a magnetic rail, the thickness dependence of a double-layer HTSC arrangement on the levitation performance was further investigated in the paper. In this study, the lower-layer bulk was polished step by step to different thicknesses, and the upper-layer bulk with constant thickness was directly superimposed on the lower-layer one. The levitation force and the force relaxation of the double-layer HTSC arrangement were measured above a Halbach magnetic rail. Experimental result shows that a bigger levitation force and a less levitation force decay could be achieved by optimizing the thickness of the lower-layer bulk HTSC. This thickness optimization method could be applied together with former reported double-layer HTSC arrangement method with aligned growth sector boundaries pattern. This series of study on the optimized combination method do bring a significant improvement on the levitation performance of present HTS maglev systems

  10. Thickness dependence of the levitation performance of double-layer high-temperature superconductor bulks above a magnetic rail

    Energy Technology Data Exchange (ETDEWEB)

    Sun, R.X.; Zheng, J.; Liao, X.L.; Che, T.; Gou, Y.F.; He, D.B.; Deng, Z.G., E-mail: zgdeng@gmail.com

    2014-10-15

    Highlights: • Thickness optimization of double-layer bulk HTSC arrangement is studied. • The new bulk HTSC arrangement makes better use of the flux distribution of the magnetic rails. • Levitation performance can be enhanced with the optimization. • The optimization can meet large levitation force requirements for HTS Maglev system. - Abstract: A double-layer high-temperature superconductor (HTSC) arrangement was proposed and proved to be able to bring improvements to both levitation force and guidance force compared with present single-layer HTSC arrangement. To fully exploit the applied magnetic field by a magnetic rail, the thickness dependence of a double-layer HTSC arrangement on the levitation performance was further investigated in the paper. In this study, the lower-layer bulk was polished step by step to different thicknesses, and the upper-layer bulk with constant thickness was directly superimposed on the lower-layer one. The levitation force and the force relaxation of the double-layer HTSC arrangement were measured above a Halbach magnetic rail. Experimental result shows that a bigger levitation force and a less levitation force decay could be achieved by optimizing the thickness of the lower-layer bulk HTSC. This thickness optimization method could be applied together with former reported double-layer HTSC arrangement method with aligned growth sector boundaries pattern. This series of study on the optimized combination method do bring a significant improvement on the levitation performance of present HTS maglev systems.

  11. Bulk Vitrification Technology For The Treatment And Immobilization Of Low-Activity Waste

    International Nuclear Information System (INIS)

    Ard, K.E.

    2011-01-01

    This report is one of four reports written to provide background information regarding immobilization technologies under consideration for supplemental immobilization of Hanford's low-activity waste. This paper is intended to provide the reader with general understanding of Bulk Vitrification and how it might be applied to immobilization of Hanford's low-activity waste.

  12. Cathepsin X Cleaves Profilin 1 C-Terminal Tyr139 and Influences Clathrin-Mediated Endocytosis.

    Directory of Open Access Journals (Sweden)

    Urša Pečar Fonović

    Full Text Available Cathepsin X, a cysteine carboxypeptidase, is upregulated in several types of cancer. Its molecular target in tumor cells is profilin 1, a known tumor suppressor and regulator of actin cytoskeleton dynamics. Cathepsin X cleaves off the C-terminal Tyr139 of profilin 1, affecting binding of poly-L-proline ligands and, consequently, tumor cell migration and invasion. Profilin 1 with mutations at the C-terminus, transiently expressed in prostate cancer cells PC-3, showed that Tyr139 is important for proper function of profilin 1 as a tumor suppressor. Cleaving off Tyr139 prevents the binding of clathrin, a poly-L-proline ligand involved in endocytosis. More profilin 1-clathrin complexes were present in PC-3 cells when cathepsin X was inhibited by its specific inhibitor AMS36 or silenced by siRNA. As a consequence, the endocytosis of FITC-labeled dextran and transferrin conjugate was significantly increased. These results constitute the first report of the regulation of clathrin-mediated endocytosis in tumor cells through proteolytic processing of profilin 1.

  13. Cathepsin X Cleaves Profilin 1 C-Terminal Tyr139 and Influences Clathrin-Mediated Endocytosis

    Science.gov (United States)

    Pečar Fonović, Urša; Kos, Janko

    2015-01-01

    Cathepsin X, a cysteine carboxypeptidase, is upregulated in several types of cancer. Its molecular target in tumor cells is profilin 1, a known tumor suppressor and regulator of actin cytoskeleton dynamics. Cathepsin X cleaves off the C-terminal Tyr139 of profilin 1, affecting binding of poly-L-proline ligands and, consequently, tumor cell migration and invasion. Profilin 1 with mutations at the C-terminus, transiently expressed in prostate cancer cells PC-3, showed that Tyr139 is important for proper function of profilin 1 as a tumor suppressor. Cleaving off Tyr139 prevents the binding of clathrin, a poly-L-proline ligand involved in endocytosis. More profilin 1—clathrin complexes were present in PC-3 cells when cathepsin X was inhibited by its specific inhibitor AMS36 or silenced by siRNA. As a consequence, the endocytosis of FITC-labeled dextran and transferrin conjugate was significantly increased. These results constitute the first report of the regulation of clathrin-mediated endocytosis in tumor cells through proteolytic processing of profilin 1. PMID:26325675

  14. Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis.

    Science.gov (United States)

    Partridge, Emily A; Le Roy, Christine; Di Guglielmo, Gianni M; Pawling, Judy; Cheung, Pam; Granovsky, Maria; Nabi, Ivan R; Wrana, Jeffrey L; Dennis, James W

    2004-10-01

    The Golgi enzyme beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is up-regulated in carcinomas and promotes the substitution of N-glycan with poly N-acetyllactosamine, the preferred ligand for galectin-3 (Gal-3). Here, we report that expression of Mgat5 sensitized mouse cells to multiple cytokines. Gal-3 cross-linked Mgat5-modified N-glycans on epidermal growth factor and transforming growth factor-beta receptors at the cell surface and delayed their removal by constitutive endocytosis. Mgat5 expression in mammary carcinoma was rate limiting for cytokine signaling and consequently for epithelial-mesenchymal transition, cell motility, and tumor metastasis. Mgat5 also promoted cytokine-mediated leukocyte signaling, phagocytosis, and extravasation in vivo. Thus, conditional regulation of N-glycan processing drives synchronous modification of cytokine receptors, which balances their surface retention against loss via endocytosis.

  15. Rab GTPases Regulate Endothelial Cell Protein C Receptor-Mediated Endocytosis and Trafficking of Factor VIIa

    Science.gov (United States)

    Nayak, Ramesh C.; Keshava, Shiva; Esmon, Charles T.; Pendurthi, Usha R.; Rao, L. Vijaya Mohan

    2013-01-01

    Recent studies have established that factor VIIa (FVIIa) binds to the endothelial cell protein C receptor (EPCR). FVIIa binding to EPCR may promote the endocytosis of this receptor/ligand complex. Rab GTPases are known to play a crucial role in the endocytic and exocytic pathways of receptors or receptor/ligand complexes. The present study was undertaken to investigate the role of Rab GTPases in the intracellular trafficking of EPCR and FVIIa. CHO-EPCR cells and human umbilical vein endothelial cells (HUVEC) were transduced with recombinant adenoviral vectors to express wild-type, constitutively active, or dominant negative mutant of various Rab GTPases. Cells were exposed to FVIIa conjugated with AF488 fluorescent probe (AF488-FVIIa), and intracellular trafficking of FVIIa, EPCR, and Rab proteins was evaluated by immunofluorescence confocal microscopy. In cells expressing wild-type or constitutively active Rab4A, internalized AF488-FVIIa accumulated in early/sorting endosomes and its entry into the recycling endosomal compartment (REC) was inhibited. Expression of constitutively active Rab5A induced large endosomal structures beneath the plasma membrane where EPCR and FVIIa accumulated. Dominant negative Rab5A inhibited the endocytosis of EPCR-FVIIa. Expression of constitutively active Rab11 resulted in retention of accumulated AF488-FVIIa in the REC, whereas expression of a dominant negative form of Rab11 led to accumulation of internalized FVIIa in the cytoplasm and prevented entry of internalized FVIIa into the REC. Expression of dominant negative Rab11 also inhibited the transport of FVIIa across the endothelium. Overall our data show that Rab GTPases regulate the internalization and intracellular trafficking of EPCR-FVIIa. PMID:23555015

  16. Biological functionalization of drug delivery carriers to bypass size restrictions of receptor-mediated endocytosis independently from receptor targeting.

    Science.gov (United States)

    Ansar, Maria; Serrano, Daniel; Papademetriou, Iason; Bhowmick, Tridib Kumar; Muro, Silvia

    2013-12-23

    Targeting of drug carriers to cell-surface receptors involved in endocytosis is commonly used for intracellular drug delivery. However, most endocytic receptors mediate uptake via clathrin or caveolar pathways associated with ≤200-nm vesicles, restricting carrier design. We recently showed that endocytosis mediated by intercellular adhesion molecule 1 (ICAM-1), which differs from clathrin- and caveolae-mediated pathways, allows uptake of nano- and microcarriers in cell culture and in vivo due to recruitment of cellular sphingomyelinases to the plasmalemma. This leads to ceramide generation at carrier binding sites and formation of actin stress-fibers, enabling engulfment and uptake of a wide size-range of carriers. Here we adapted this paradigm to enhance uptake of drug carriers targeted to receptors associated with size-restricted pathways. We coated sphingomyelinase onto model (polystyrene) submicro- and microcarriers targeted to clathrin-associated mannose-6-phosphate receptor. In endothelial cells, this provided ceramide enrichment at the cell surface and actin stress-fiber formation, modifying the uptake pathway and enhancing carrier endocytosis without affecting targeting, endosomal transport, cell-associated degradation, or cell viability. This improvement depended on the carrier size and enzyme dose, and similar results were observed for other receptors (transferrin receptor) and cell types (epithelial cells). This phenomenon also enhanced tissue accumulation of carriers after intravenous injection in mice. Hence, it is possible to maintain targeting toward a selected receptor while bypassing natural size restrictions of its associated endocytic route by functionalization of drug carriers with biological elements mimicking the ICAM-1 pathway. This strategy holds considerable promise to enhance flexibility of design of targeted drug delivery systems.

  17. BULK VITRIFICATION TECHNOLOGY FOR THE TREATMENT AND IMMOBILIZATION OF LOW-ACTIVITY WASTE

    Energy Technology Data Exchange (ETDEWEB)

    ARD KE

    2011-04-11

    This report is one of four reports written to provide background information regarding immobilization technologies under consideration for supplemental immobilization of Hanford's low-activity waste. This paper is intended to provide the reader with general understanding of Bulk Vitrification and how it might be applied to immobilization of Hanford's low-activity waste.

  18. Endocytosis of the major yolk proteins of the silkmoth, Hyalophora cecropia: Uptake kinetics and interactions

    International Nuclear Information System (INIS)

    Kulakosky, P.C.

    1989-01-01

    The oocytes of Lepidopteran insects take up several yolk proteins in defined proportions even though their relative availability in the hemolymph changes during the several days required to complete yolk formation in all the eggs. There are three hemolymph yolk precursors, vitellogenin, microvitellogenin and lipophorin; one precursor, paravitellogenin is produced in the ovary. The control mechanism for their proportional endocytosis is not known. In this thesis, the author describe the purification of all four proteins and the radiolabeling of the hemolymph precursors. The radiolabeled proteins were tested with an in vitro incubation system to assess the biological activity of the proteins and the reliability of the incubation methods. All of the labeled probes were transferred from the incubation medium to yolk spheres within the oocyte in a saturable, energy-dependent, and stage-specific manner. The rates of uptake were similar to the estimated rates of uptake in situ. The concentration dependence of in vitro uptake was investigated and found to be consistent with in situ concentrations and the composition of yolk in mature eggs. Two precursors, vitellogenin and lipophorin, competed for uptake indicating that they share a common binding site while the third, microvitellin, did not compete with the others. Though vitellogenin and lipophorin competed for uptake, only vitellogenin displayed the unique ability to increase the uptake rate of microvitellin and fluid in vitro

  19. Endocytosis of a maltose permease is induced when amylolytic enzyme production is repressed in Aspergillus oryzae.

    Science.gov (United States)

    Hiramoto, Tetsuya; Tanaka, Mizuki; Ichikawa, Takanori; Matsuura, Yuka; Hasegawa-Shiro, Sachiko; Shintani, Takahiro; Gomi, Katsuya

    2015-09-01

    In the filamentous fungus Aspergillus oryzae, amylolytic enzyme production is induced by the presence of maltose. Previously, we identified a putative maltose permease (MalP) gene in the maltose-utilizing cluster of A. oryzae. malP disruption causes a significant decrease in α-amylase activity and maltose consumption, indicating that MalP is a maltose transporter required for amylolytic enzyme production in A. oryzae. Although the expression of amylase genes and malP is repressed by the presence of glucose, the effect of glucose on the abundance of functional MalP is unknown. In this study, we examined the effect of glucose and other carbon sources on the subcellular localization of green fluorescence protein (GFP)-tagged MalP. After glucose addition, GFP-MalP at the plasma membrane was internalized and delivered to the vacuole. This glucose-induced internalization of GFP-MalP was inhibited by treatment with latrunculin B, an inhibitor of actin polymerization. Furthermore, GFP-MalP internalization was inhibited by repressing the HECT ubiquitin ligase HulA (ortholog of yeast Rsp5). These results suggest that MalP is transported to the vacuole by endocytosis in the presence of glucose. Besides glucose, mannose and 2-deoxyglucose also induced the endocytosis of GFP-MalP and amylolytic enzyme production was inhibited by the addition of these sugars. However, neither the subcellular localization of GFP-MalP nor amylolytic enzyme production was influenced by the addition of xylose or 3-O-methylglucose. These results imply that MalP endocytosis is induced when amylolytic enzyme production is repressed. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. MUC1 intra-cellular trafficking is clathrin, dynamin, and rab5 dependent

    International Nuclear Information System (INIS)

    Liu Xiaolong; Yuan Zhenglong; Chung, Maureen

    2008-01-01

    MUC1, a transmembrane glycoprotein, is abnormally over-expressed in most human adenocarcinomas. MUC1 association with cytoplasmic cell signal regulators and nuclear accumulation are important for its tumor related activities. Little is known about how MUC1 translocates from the cell membrane to the cytoplasm. In this study, live cell imaging was used to study MUC1 intracellular trafficking. The interaction between EGFR and MUC1 was mapped by FRET analysis and EGF stimulated MUC1 endocytosis was observed directly through live cell imaging. MUC1-CT endocytosis was clathrin and dynamin dependent. Rab5 over-expression resulted in decreased cell membrane localization of MUC1, with accumulation of MUC1 endocytic vesicles in the peri-nuclear region. Conversely, over-expression of a Rab5 dominant negative mutant (S34N) resulted in redistribution of MUC1 from the peri-nuclear region to the cytoplasm. Collectively, these results indicated that MUC1 intra-cellular trafficking occurs through a regulated process that was stimulated by direct EGFR and MUC1 interaction, mediated by clathrin coated pits that were dynamin dependent and regulated by Rab5

  1. Endocytosis and exocytosis of nanoparticles in mammalian cells

    OpenAIRE

    Park, Ji- Ho; Oh,Nuri

    2014-01-01

    Nuri Oh,1,2 Ji-Ho Park1–31Department of Bio and Brain Engineering, 2Institute for Optical Science and Technology, 3Institute for the NanoCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of KoreaAbstract: Engineered nanoparticles that can be injected into the human body hold tremendous potential to detect and treat complex diseases. Understanding of the endocytosis and exocytosis mechanisms of nanoparticles is essential for safe and efficient the...

  2. Optimisation of amylase and xylanase addition in dependance of white flour amylase activity

    Directory of Open Access Journals (Sweden)

    Lončar Davor M.

    2016-01-01

    Full Text Available In this study the effect of different quantities of added amylase to white wheat flours characterized with different activities of naturally existing amylases is tested. Response surface methodology is chosen to test the effects of main applied technological parameters on bread quality responses. Independent variables are chosen to be: quantity of added amylase and bulk fermentation time, while analysed responses are: specific volume, grain structure, bulk fermentation. Bread quality responses are statistically significant, while predicted and observed responses correspond very well, which allows good prediction of bread quality parameters based on applied technological parameters and flour characteristics. Score analysis shows that optimum quantity of amylase addition regarding bread quality depends on the activity of naturally existing amylases. Optimal quantity of added xylanase in bread samples made from both flour types is 0.004%. Xylanase improved properties of white wheat bread and higher effect is experienced with flour that has more active naturally existing amylases. Addition of amylase has statistically significantly increased a* values of crust. Addition of xylanase has statistically significantly decreased values of b* in comparison to the respective bread sample with only added amylase.

  3. Exocytosis and endocytosis in juxtaglomerular cells

    DEFF Research Database (Denmark)

    Friis, U G; Jensen, B L; Hansen, Pernille B. Lærkegaard

    2000-01-01

    fusion events between secretory granules and cell membrane and measurement of intermittent secretion of renin from single afferent arterioles, with a renin content of each secretion episode that corresponds to the renin content of one secretory granule. More recently it has been demonstrated...... that the afferent arterioles lose a large number of renin granules after acute stimulation without changing the average granular volume. Current electrophysiological techniques have now permitted direct measurements of cell membrane capacitance in juxtaglomerular (JG) cells as a measure of net addition (exocytosis...... and endocytosis are regulated processes in the JG-cells and both may be important for the long-term control of renin secretion at the single cell level....

  4. Dissecting Bacterial Cell Wall Entry and Signaling in Eukaryotic Cells: an Actin-Dependent Pathway Parallels Platelet-Activating Factor Receptor-Mediated Endocytosis.

    Science.gov (United States)

    Loh, Lip Nam; Gao, Geli; Tuomanen, Elaine I

    2017-01-03

    The Gram-positive bacterial cell wall (CW) peptidoglycan-teichoic acid complex is released into the host environment during bacterial metabolism or death. It is a highly inflammatory Toll-like receptor 2 (TLR2) ligand, and previous in vivo studies have demonstrated its ability to recapitulate pathological features of pneumonia and meningitis. We report that an actin-dependent pathway is involved in the internalization of the CW by epithelial and endothelial cells, in addition to the previously described platelet-activating factor receptor (PAFr)-dependent uptake pathway. Unlike the PAFr-dependent pathway, which is mediated by clathrin and dynamin and does not lead to signaling, the alternative pathway is sensitive to 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and engenders Rac1, Cdc42, and phosphatidylinositol 3-kinase (PI3K) signaling. Upon internalization by this macropinocytosis-like pathway, CW is trafficked to lysosomes. Intracellular CW trafficking is more complex than previously recognized and suggests multiple points of interaction with and without innate immune signaling. Streptococcus pneumoniae is a major human pathogen infecting the respiratory tract and brain. It is an established model organism for understanding how infection injures the host. During infection or bacterial growth, bacteria shed their cell wall (CW) into the host environment and trigger inflammation. A previous study has shown that CW enters and crosses cell barriers by interacting with a receptor on the surfaces of host cells, termed platelet-activating factor receptor (PAFr). In the present study, by using cells that are depleted of PAFr, we identified a second pathway with features of macropinocytosis, which is a receptor-independent fluid uptake mechanism by cells. Each pathway contributes approximately the same amount of cell wall trafficking, but the PAFr pathway is silent, while the new pathway appears to contribute to the host inflammatory response to CW insult. Copyright © 2017

  5. Centrality dependence of bulk fireball properties in √(sNN)=200 GeV Au-Au collisions

    International Nuclear Information System (INIS)

    Rafelski, Johann; Letessier, Jean; Torrieri, Giorgio

    2005-01-01

    We explore the centrality dependence of the properties of the dense hadronic matter created in √(s NN )=200 GeV Au-Au collisions at the Relativistic Heavy Ion Collider. Using the statistical hadronization model, we fit particle yields known for 11 centrality bins. We present the resulting model parameters, rapidity yields of physical quantities, and the physical properties of bulk matter at hadronization as function of centrality. We discuss the production of strangeness and entropy

  6. Perfluorooctanoic acid affects endocytosis involving clathrin light chain A and microRNA-133b-3p in mouse testes

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Yin; Wang, Jianshe [Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101 (China); Guo, Xuejiang [State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, PR China. (China); Yan, Shengmin [Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101 (China); Dai, Jiayin, E-mail: daijy@ioz.ac.cn [Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101 (China)

    2017-03-01

    Perfluorooctanoic acid (PFOA) is an abundant perfluoroalkyl substance widely applied in industrial and consumer products. Among its potential health hazards, testicular toxicity is of major concern. To explore the potential effect of miRNA on post-translational regulation after PFOA exposure, changes in miRNAs were detected via miRNA array. Seventeen miRNAs were differentially expressed (eight upregulated, nine downregulated) in male mouse testes after exposure to 5 mg/kg/d of PFOA for 28 d (> 1.5-fold and P < 0.05 compared with the control). Eight of these miRNAs were further selected for TaqMan qPCR analysis. Proteomic profile analysis indicated that many changed proteins after PFOA treatment, including intersectin 1 (ITSN1), serine protease inhibitor A3K (Serpina3k), and apolipoprotein a1 (APOA1), were involved in endocytosis and blood-testis barrier (BTB) processes. These changes were further verified by immunohistochemical and Western blot analyses. Endocytosis-related genes were selected for qPCR analysis, with many found to be significantly changed after PFOA treatment, including epidermal growth factor receptor pathway substrate 8 (Eps8), Eps15, cortactin, cofilin, espin, vinculin, and zyxin. We further predicted the potential interaction between changed miRNAs and proteins, which indicated that miRNAs might play a role in the post-translational regulation of gene expression after PFOA treatment in mouse testes. Among them, miR-133b-3p/clathrin light chain A (CLTA) was selected and verified in vitro by transfection and luciferase activity assay. Results showed that PFOA exposure affects endocytosis in mouse testes and that CLTA is a potential target of miR-133b-3p. - Highlights: • Endocytosis and blood-testis barrier proteins were changed after PFOA exposure. • Seventeen miRNAs were differentially expressed in testes after PFOA exposure. • MiRNAs might play a role in gene regulation in testes after PFOA exposure.CLTA is a potential target of miR-133b

  7. Perfluorooctanoic acid affects endocytosis involving clathrin light chain A and microRNA-133b-3p in mouse testes

    International Nuclear Information System (INIS)

    Lu, Yin; Wang, Jianshe; Guo, Xuejiang; Yan, Shengmin; Dai, Jiayin

    2017-01-01

    Perfluorooctanoic acid (PFOA) is an abundant perfluoroalkyl substance widely applied in industrial and consumer products. Among its potential health hazards, testicular toxicity is of major concern. To explore the potential effect of miRNA on post-translational regulation after PFOA exposure, changes in miRNAs were detected via miRNA array. Seventeen miRNAs were differentially expressed (eight upregulated, nine downregulated) in male mouse testes after exposure to 5 mg/kg/d of PFOA for 28 d (> 1.5-fold and P < 0.05 compared with the control). Eight of these miRNAs were further selected for TaqMan qPCR analysis. Proteomic profile analysis indicated that many changed proteins after PFOA treatment, including intersectin 1 (ITSN1), serine protease inhibitor A3K (Serpina3k), and apolipoprotein a1 (APOA1), were involved in endocytosis and blood-testis barrier (BTB) processes. These changes were further verified by immunohistochemical and Western blot analyses. Endocytosis-related genes were selected for qPCR analysis, with many found to be significantly changed after PFOA treatment, including epidermal growth factor receptor pathway substrate 8 (Eps8), Eps15, cortactin, cofilin, espin, vinculin, and zyxin. We further predicted the potential interaction between changed miRNAs and proteins, which indicated that miRNAs might play a role in the post-translational regulation of gene expression after PFOA treatment in mouse testes. Among them, miR-133b-3p/clathrin light chain A (CLTA) was selected and verified in vitro by transfection and luciferase activity assay. Results showed that PFOA exposure affects endocytosis in mouse testes and that CLTA is a potential target of miR-133b-3p. - Highlights: • Endocytosis and blood-testis barrier proteins were changed after PFOA exposure. • Seventeen miRNAs were differentially expressed in testes after PFOA exposure. • MiRNAs might play a role in gene regulation in testes after PFOA exposure.CLTA is a potential target of miR-133b

  8. Endocytosis Pathways of the Folate Tethered Star-Shaped PEG-PCL Micelles in Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Yu-Lun Li

    2014-03-01

    Full Text Available This study reports on the cellular uptake of folate tethered micelles using a branched skeleton of poly(ethylene glycol and poly(ε-caprolactone. The chemical structures of the copolymers were characterized by proton nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. Doxorubicin (DOX was utilized as an anticancer drug. The highest drug loading efficiencies of DOX in the folate decorated micelle (DMCF and folate-free micelle (DMC were found to be 88.5% and 88.2%, respectively, depending on the segment length of the poly(ε-caprolactone in the copolymers. A comparison of fluorescent microscopic images of the endocytosis pathway in two cell lines, human breast cancer cells (MCF-7 and human oral cavity carcinoma cells (KB, revealed that the micelles were engulfed by KB and MCF-7 cells following in vitro incubation for one hour. Flow cytometric analysis revealed that free folic acid can inhibit the uptake of DOX by 48%–57% and 26%–39% in KB cells and MCF-7 cells, respectively. These results prove that KB cells are relatively sensitive to folate-tethered micelles. Upon administering methyl-β-cyclodextrin, an inhibitor of the caveolae-mediated endocytosis pathway, the uptake of DOX by KB cells was reduced by 69% and that by MCF-7 cells was reduced by 56%. This finding suggests that DMCF enters cells via multiple pathways, thus implying that the folate receptor is not the only target of tumor therapeutics.

  9. Ricin A chain reaches the endoplasmic reticulum after endocytosis

    International Nuclear Information System (INIS)

    Liu Qiong; Zhan Jinbiao; Chen Xinhong; Zheng Shu

    2006-01-01

    Ricin is a potent ribosome inactivating protein and now has been widely used for synthesis of immunotoxins. To target ribosome in the mammalian cytosol, ricin must firstly retrograde transport from the endomembrane system to reach the endoplasmic reticulum (ER) where the ricin A chain (RTA) is recognized by ER components that facilitate its membrane translocation to the cytosol. In the study, the fusion gene of enhanced green fluorescent protein (EGFP)-RTA was expressed with the pET-28a (+) system in Escherichia coli under the control of a T7 promoter. The fusion protein showed a green fluorescence. The recombinant protein can be purified by metal chelated affinity chromatography on a column of NTA. The rabbit anti-GFP antibody can recognize the fusion protein of EGFP-RTA just like the EGFP protein. The cytotoxicity of EGFP-RTA and RTA was evaluated by the MTT assay in HeLa and HEP-G2 cells following fluid-phase endocytosis. The fusion protein had a similar cytotoxicity of RTA. After endocytosis, the subcellular location of the fusion protein can be observed with the laser scanning confocal microscopy and the immuno-gold labeling Electro Microscopy. This study provided important evidence by a visualized way to prove that RTA does reach the endoplasmic reticulum

  10. Understanding the effect of surface/bulk defects on the photocatalytic activity of TiO2: anatase versus rutile.

    Science.gov (United States)

    Yan, Junqing; Wu, Guangjun; Guan, Naijia; Li, Landong; Li, Zhuoxin; Cao, Xingzhong

    2013-07-14

    The sole effect of surface/bulk defects of TiO2 samples on their photocatalytic activity was investigated. Nano-sized anatase and rutile TiO2 were prepared by hydrothermal method and their surface/bulk defects were adjusted simply by calcination at different temperatures, i.e. 400-700 °C. High temperature calcinations induced the growth of crystalline sizes and a decrease in the surface areas, while the crystalline phase and the exposed facets were kept unchanged during calcination, as indicated by the characterization results from XRD, Raman, nitrogen adsorption-desorption, TEM and UV-Vis spectra. The existence of surface/bulk defects in calcined TiO2 samples was confirmed by photoluminescence and XPS spectra, and the surface/bulk defect ratio was quantitatively analyzed according to positron annihilation results. The photocatalytic activity of calcined TiO2 samples was evaluated in the photocatalytic reforming of methanol and the photocatalytic oxidation of α-phenethyl alcohol. Based on the characterization and catalytic results, a direct correlation between the surface specific photocatalytic activity and the surface/bulk defect density ratio could be drawn for both anatase TiO2 and rutile TiO2. The surface defects of TiO2, i.e. oxygen vacancy clusters, could promote the separation of electron-hole pairs under irradiation, and therefore, enhance the activity during photocatalytic reaction.

  11. Caveolae-mediated endocytosis of biocompatible gold nanoparticles in living Hela cells

    DEFF Research Database (Denmark)

    Hao, Xian; Wu, Jiazhen; Shan, Yuping

    2012-01-01

    the internalization mechanism of small-size AuNPs by living Hela cells. Herein, we found that the caveolae-mediated endocytosis was the dominant pathway for the intracellular delivery of small-size AuNPs. The intracellular delivery was suppressed when we depleted the cholesterol with methyl-β-cyclodextrin (M beta CD...

  12. State dependent arrival in bulk retrial queueing system with immediate Bernoulli feedback, multiple vacations and threshold

    Science.gov (United States)

    Niranjan, S. P.; Chandrasekaran, V. M.; Indhira, K.

    2017-11-01

    The objective of this paper is to analyse state dependent arrival in bulk retrial queueing system with immediate Bernoulli feedback, multiple vacations, threshold and constant retrial policy. Primary customers are arriving into the system in bulk with different arrival rates λ a and λ b . If arriving customers find the server is busy then the entire batch will join to orbit. Customer from orbit request service one by one with constant retrial rate γ. On the other hand if an arrival of customers finds the server is idle then customers will be served in batches according to general bulk service rule. After service completion, customers may request service again with probability δ as feedback or leave from the system with probability 1 - δ. In the service completion epoch, if the orbit size is zero then the server leaves for multiple vacations. The server continues the vacation until the orbit size reaches the value ‘N’ (N > b). At the vacation completion, if the orbit size is ‘N’ then the server becomes ready to provide service for customers from the main pool or from the orbit. For the designed queueing model, probability generating function of the queue size at an arbitrary time will be obtained by using supplementary variable technique. Various performance measures will be derived with suitable numerical illustrations.

  13. Bulk - Samples gamma-rays activation analysis (PGNAA) with Isotopic Neutron Sources

    International Nuclear Information System (INIS)

    HASSAN, A.M.

    2009-01-01

    An overview is given on research towards the Prompt Gamma-ray Neutron Activation Analysis (PGNAA) of bulk-samples. Some aspects in bulk-sample PGNAA are discussed, where irradiation by isotopic neutron sources is used mostly for in-situ or on-line analysis. The research was carried out in a comparative and/or qualitative way or by using a prior knowledge about the sample material. Sometimes we need to use the assumption that the mass fractions of all determined elements add up to 1. The sensitivity curves are also used for some elements in such complex samples, just to estimate the exact percentage concentration values. The uses of 252 Cf, 241 Arn/Be and 239 Pu/Be isotopic neutron sources for elemental investigation of: hematite, ilmenite, coal, petroleum, edible oils, phosphates and pollutant lake water samples have been mentioned.

  14. Inhibition of endocytosis blocks Wnt signalling to beta-catenin by promoting dishevelled degradation

    Czech Academy of Sciences Publication Activity Database

    Bryja, Vítězslav; Čajánek, L.; Grahn, A.; Schulte, G.

    2007-01-01

    Roč. 190, č. 1 (2007), s. 53-596 ISSN 0001-6772 Institutional research plan: CEZ:AV0Z50040507 Keywords : beta-catenin * clathrin-mediated endocytosis * desensitization Subject RIV: BO - Biophysics Impact factor: 2.554, year: 2007

  15. Understanding magnetic nanoparticle osteoblast receptor-mediated endocytosis using experiments and modeling

    International Nuclear Information System (INIS)

    Tran, Nhiem; Webster, Thomas J

    2013-01-01

    Iron oxide nanoparticles are promising candidates for controlling drug delivery through an external magnetic force to treat a wide range of diseases, including osteoporosis. Previous studies have demonstrated that in the presence of hydroxyapatite coated magnetite (Fe 3 O 4 ) nanoparticles, osteoblast (or bone forming cell) proliferation and long-term functions (such as calcium deposition) were significantly enhanced. Hydroxyapatite is the major inorganic component of bone. As a further attempt to understand why, in the current study, the uptake of such nanoparticles into osteoblasts was experimentally investigated and mathematically modeled. Magnetite nanoparticles were synthesized using a co-precipitation method and were coated with hydroxyapatite. A cellular uptake experiment at low temperatures indicated that receptor-mediated endocytosis contributed to the internalization of the magnetic nanoparticles into osteoblasts. A model was further developed to explain the uptake of magnetic nanoparticles into osteoblasts using receptor-mediated endocytosis. This model may explain the internalization of hydroxyapatite into osteoblasts to elevate intracellular calcium levels necessary to promote osteoblast functions to treat a wide range of orthopedic problems, including osteoporosis. (paper)

  16. Bulk viscosity of spin-one color superconductors

    Energy Technology Data Exchange (ETDEWEB)

    Sa' d, Basil A.

    2009-08-27

    The bulk viscosity of several quark matter phases is calculated. It is found that the effect of color superconductivity is not trivial, it may suppress, or enhance the bulk viscosity depending on the critical temperature and the temperature at which the bulk viscosity is calculated. Also, is it found that the effect of neutrino-emitting Urca processes cannot be neglected in the consideration of the bulk viscosity of strange quark matter. The results for the bulk viscosity of strange quark matter are used to calculate the r-mode instability window of quark stars with several possible phases. It is shown that each possible phase has a different structure for the r-mode instability window. (orig.)

  17. Bulk viscosity of spin-one color superconductors

    International Nuclear Information System (INIS)

    Sa'd, Basil A.

    2009-01-01

    The bulk viscosity of several quark matter phases is calculated. It is found that the effect of color superconductivity is not trivial, it may suppress, or enhance the bulk viscosity depending on the critical temperature and the temperature at which the bulk viscosity is calculated. Also, is it found that the effect of neutrino-emitting Urca processes cannot be neglected in the consideration of the bulk viscosity of strange quark matter. The results for the bulk viscosity of strange quark matter are used to calculate the r-mode instability window of quark stars with several possible phases. It is shown that each possible phase has a different structure for the r-mode instability window. (orig.)

  18. NtGNL1a ARF-GEF acts in endocytosis in tobacco cells

    Czech Academy of Sciences Publication Activity Database

    Jelínková, Adriana; Müller, Karel; Pařezová, Markéta; Petrášek, Jan

    2015-01-01

    Roč. 15, NOV 5 (2015), s. 272 ISSN 1471-2229 R&D Projects: GA ČR GPP305/11/P797 Institutional support: RVO:61389030 Keywords : Endocytosis * PIN1 protein trafficking * Inhibitors of endomembrane trafficking Subject RIV: EA - Cell Biology Impact factor: 3.631, year: 2015

  19. Detergent insolubility of alkaline phosphatase during biosynthetic transport and endocytosis. Role of cholesterol

    NARCIS (Netherlands)

    Cerneus, D. P.; Ueffing, E.; Posthuma, G.; Strous, G. J.; van der Ende, A.

    1993-01-01

    Alkaline phosphatase is anchored to the outer leaflet of the plasma membrane by a covalently attached glycosyl-phosphatidylinositol anchor. We have studied the biosynthetic transport and endocytosis of alkaline phosphatase in the choriocarcinoma cell line BeWo, which endogenously expresses this

  20. Endocytic crosstalk: cavins, caveolins, and caveolae regulate clathrin-independent endocytosis.

    Directory of Open Access Journals (Sweden)

    Natasha Chaudhary

    2014-04-01

    Full Text Available Several studies have suggested crosstalk between different clathrin-independent endocytic pathways. However, the molecular mechanisms and functional relevance of these interactions are unclear. Caveolins and cavins are crucial components of caveolae, specialized microdomains that also constitute an endocytic route. Here we show that specific caveolar proteins are independently acting negative regulators of clathrin-independent endocytosis. Cavin-1 and Cavin-3, but not Cavin-2 or Cavin-4, are potent inhibitors of the clathrin-independent carriers/GPI-AP enriched early endosomal compartment (CLIC/GEEC endocytic pathway, in a process independent of caveola formation. Caveolin-1 (CAV1 and CAV3 also inhibit the CLIC/GEEC pathway upon over-expression. Expression of caveolar protein leads to reduction in formation of early CLIC/GEEC carriers, as detected by quantitative electron microscopy analysis. Furthermore, the CLIC/GEEC pathway is upregulated in cells lacking CAV1/Cavin-1 or with reduced expression of Cavin-1 and Cavin-3. Inhibition by caveolins can be mimicked by the isolated caveolin scaffolding domain and is associated with perturbed diffusion of lipid microdomain components, as revealed by fluorescence recovery after photobleaching (FRAP studies. In the absence of cavins (and caveolae CAV1 is itself endocytosed preferentially through the CLIC/GEEC pathway, but the pathway loses polarization and sorting attributes with consequences for membrane dynamics and endocytic polarization in migrating cells and adult muscle tissue. We also found that noncaveolar Cavin-1 can act as a modulator for the activity of the key regulator of the CLIC/GEEC pathway, Cdc42. This work provides new insights into the regulation of noncaveolar clathrin-independent endocytosis by specific caveolar proteins, illustrating multiple levels of crosstalk between these pathways. We show for the first time a role for specific cavins in regulating the CLIC/GEEC pathway, provide

  1. Electronic relaxation of deep bulk trap and interface state in ZnO ceramics

    International Nuclear Information System (INIS)

    Yang Yan; Li Sheng-Tao; Ding Can; Cheng Peng-Fei

    2011-01-01

    This paper investigates the electronic relaxation of deep bulk trap and interface state in ZnO ceramics based on dielectric spectra measured in a wide range of temperature, frequency and bias, in addition to the steady state response. It discusses the nature of net current flowing over the barrier affected by interface state, and then obtains temperature-dependent barrier height by approximate calculation from steady I—V (current—voltage) characteristics. Additional conductance and capacitance arising from deep bulk trap relaxation are calculated based on the displacement of the cross point between deep bulk trap and Fermi level under small AC signal. From the resonances due to deep bulk trap relaxation on dielectric spectra, the activation energies are obtained as 0.22 eV and 0.35 eV, which are consistent with the electronic levels of the main defect interstitial Zn and vacancy oxygen in the depletion layer. Under moderate bias, another resonance due to interface relaxation is shown on the dielectric spectra. The DC-like conductance is also observed in high temperature region on dielectric spectra, and the activation energy is much smaller than the barrier height in steady state condition, which is attributed to the displacement current coming from the shallow bulk trap relaxation or other factors. (fluids, plasmas and electric discharges)

  2. Magnetic field dependence of vortex activation energy

    Indian Academy of Sciences (India)

    ... the resistance as a function of temperature and magnetic field in clean polycrystalline samples of NbSe2, MgB2 and Bi2Sr2Ca2Cu3O10 (BSCCO) superconductors. Thermally activated flux flow behaviour is seen in all the three systems and clearly identified in bulk MgB2. While the activation energy at low fields for MgB2 ...

  3. Analytical Model for Voltage-Dependent Photo and Dark Currents in Bulk Heterojunction Organic Solar Cells

    Directory of Open Access Journals (Sweden)

    Mesbahus Saleheen

    2016-05-01

    Full Text Available A physics-based explicit mathematical model for the external voltage-dependent forward dark current in bulk heterojunction (BHJ organic solar cells is developed by considering Shockley-Read-Hall (SRH recombination and solving the continuity equations for both electrons and holes. An analytical model for the external voltage-dependent photocurrent in BHJ organic solar cells is also proposed by incorporating exponential photon absorption, dissociation efficiency of bound electron-hole pairs (EHPs, carrier trapping, and carrier drift and diffusion in the photon absorption layer. Modified Braun’s model is used to compute the electric field-dependent dissociation efficiency of the bound EHPs. The overall net current is calculated considering the actual solar spectrum. The mathematical models are verified by comparing the model calculations with various published experimental results. We analyze the effects of the contact properties, blend compositions, charge carrier transport properties (carrier mobility and lifetime, and cell design on the current-voltage characteristics. The power conversion efficiency of BHJ organic solar cells mostly depends on electron transport properties of the acceptor layer. The results of this paper indicate that improvement of charge carrier transport (both mobility and lifetime and dissociation of bound EHPs in organic blend are critically important to increase the power conversion efficiency of the BHJ solar cells.

  4. Stress- and temperature-dependent scaling behavior of dynamic hysteresis in soft PZT bulk ceramics

    International Nuclear Information System (INIS)

    Yimnirun, R; Wongsaenmai, S; Wongmaneerung, R; Wongdamnern, N; Ngamjarurojana, A; Ananta, S; Laosiritaworn, Y

    2007-01-01

    Effects of electric field-frequency, electric field-amplitude, mechanical stress, and temperature on the hysteresis area, especially the scaling form, were investigated in soft lead zirconate titanate (PZT) bulk ceramics. The hysteresis area was found to depend on the frequency and field-amplitude with the same set of exponents as the power-law scaling for both with and without stresses. The inclusion of stresses into the power-law was obtained in the form of σ=0 > ∝ f -0.25 E 0 σ 0.45 which indicates the difference in energy dissipation between the under-stress and stress-free conditions. The power-law temperature scaling relations were obtained for hysteresis area (A) and remanent polarization P r , while the coercivity E C was found to scale linearly with temperature T. The three temperature scaling relations were also field-dependent. At fixed field amplitude E 0 , the scaling relations take the forms of ∝ T -1.1024 , P r ∼T -1.2322 and (E C0 - E C ) ∼T

  5. Intestinal alkaline phosphatase: selective endocytosis from the enterocyte brush border during fat absorption

    DEFF Research Database (Denmark)

    Hansen, Gert Helge; Niels-Christiansen, Lise-Lotte; Immerdal, Lissi

    2007-01-01

    explants. By immunofluorescence microscopy, fat absorption caused a translocation of IAP from the enterocyte brush border to the interior of the cell, whereas other brush-border enzymes were unaffected. By electron microscopy, the translocation occurred by a rapid (5 min) induction of endocytosis via...

  6. Clathrin-mediated endocytosis and transcytosis of enterotoxigenic Escherichia coli F4 fimbriae in porcine intestinal epithelial cells.

    Science.gov (United States)

    Rasschaert, Kristien; Devriendt, Bert; Favoreel, Herman; Goddeeris, Bruno M; Cox, Eric

    2010-10-15

    Enterotoxigenic Escherichia coli (ETEC) cause severe diarrhea in neonatal and recently weaned piglets. Previously, we demonstrated that oral immunization of F4 receptor positive piglets with purified F4 fimbriae induces a protective F4-specific intestinal immune response. However, in F4 receptor negative animals no F4-specific immune response can be elicited, indicating that the induction of an F4-specific mucosal immune response upon oral immunisation is receptor-dependent. Although F4 fimbriae undergo transcytosis across the intestinal epithelium in vivo, the endocytosis pathways used remain unknown. In the present study, we characterized the internalization of F4 fimbriae in the porcine intestinal epithelial cell line IPEC-J2. The results in the present study demonstrate that F4 fimbriae are internalized through a clathrin-dependent pathway. Furthermore, our results suggest that F4 fimbriae are transcytosed across differentiated IPEC-J2 cells. This receptor-dependent transcytosis of F4 fimbriae may explain the immunogenicity of these fimbriae upon oral administration in vivo. (c) 2010 Elsevier B.V. All rights reserved.

  7. AMPA Receptor Endocytosis in Rat Perirhinal Cortex Underlies Retrieval of Object Memory

    Science.gov (United States)

    Cazakoff, Brittany N.; Howland, John G.

    2011-01-01

    Mechanisms consistent with long-term depression in the perirhinal cortex (PRh) play a fundamental role in object recognition memory; however, whether AMPA receptor endocytosis is involved in distinct phases of recognition memory is not known. To address this question, we used local PRh infusions of the cell membrane-permeable Tat-GluA2[subscript…

  8. Theory of bulk-surface coupling in topological insulator films

    Science.gov (United States)

    Saha, Kush; Garate, Ion

    2014-12-01

    We present a quantitative microscopic theory of the disorder- and phonon-induced coupling between surface and bulk states in doped topological insulator films. We find a simple mathematical structure for the surface-to-bulk scattering matrix elements and confirm the importance of bulk-surface coupling in transport and photoemission experiments, assessing its dependence on temperature, carrier density, film thickness, and particle-hole asymmetry.

  9. Synthesis of formamidinium lead iodide perovskite bulk single crystal and its optical properties

    Science.gov (United States)

    Zheng, Hongge; Duan, Junjie; Dai, Jun

    2017-07-01

    Formamidinium lead iodide (FAPbI3) is a promising hybrid perovskite material for optoelectronic devices. We synthesized bulk single crystal FAPbI3 by a rapid solution crystallization method. X-ray diffraction (XRD) was performed to characterize the crystal structure. Temperature-dependent photoluminescence (PL) spectra of the bulk single crystal FAPbI3 were measured from 10 to 300 K to explain PL recombination mechanism. It shows that near band edge emission blueshifts with the temperature increasing from 10 to 120 K and from 140 K to room temperature, a sudden emission band redshift demonstrates near 140 K because of the phase transition from orthorhombic phase to cubic phase. From the temperature-dependent PL spectra, the temperature coefficients of the bandgap and thermal activation energies of FAPbI3 perovskite are fitted.

  10. Stochastic Modeling of the Clathrin-dependent and -independent Endocytic Pathways

    Science.gov (United States)

    Deng, Hua; Dutta, Prashanta; Liu, Jin

    2017-11-01

    Endocytosis is one of the important processes that bioparticles use to enter the cells. During endocytosis the membrane-bound vesicles are formed by the invagination of plasma membrane as a result of interactions among many proteins and cytoskeletons. The clathrin-mediated endocytosis is one of the most significant form of endocytosis, where the dynamic assembly of clathrin-coated pits play a critical role. While herpes simplex virus-1 has recently shown to infect cell by a novel phagocytosis-like endocytic pathway where actin polymerization may facilitate the viral entry. In this work, we propose a stochastic model for both clathrin-dependent and -independent endocytic pathways based on Monte Carlo simulations. The important roles of clathrin coating and actin cytoskeleton as well as the impact of other biological parameters are studied. Our preliminary results indicate that there exist an intermediate particle size and ligand density that maximize the internalization efficiency. Below a critical size or surface ligand density, it is difficult for the entry of a single particle, which means clustering may needed for more efficient internalization. We also find that lower membrane bending rigidity may help promote the bioparticle entry. Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R01GM122081.

  11. Spatiotemporal analysis of endocytosis and membrane distribution of fluorescent sterols in living cells

    DEFF Research Database (Denmark)

    Wüstner, Daniel; Faergeman, Nils J

    2008-01-01

    proximity to the cell membrane. Spatial surface intensity patterns of DHE as well as that of the lipid marker DiIC12 being assessed by statistical image analysis persisted over several minutes in cells having a constant overall curvature. Sites of sterol endocytosis appeared indistinguishable from other...

  12. The effect of vanadate on receptor-mediated endocytosis of asialoorosomucoid in rat liver parenchymal cells

    International Nuclear Information System (INIS)

    Kindberg, G.M.; Gudmundsen, O.; Berg, T.

    1990-01-01

    Vanadate is a phosphate analogue that inhibits enzymes involved in phosphate release and transfer reactions. Since such reactions may play important roles in endocytosis, we studied the effects of vanadate on various steps in receptor-mediated endocytosis of asialoorosomucoid labeled with 125I-tyramine-cellobiose (125I-TC-AOM). The labeled degradation products formed from 125I-TC-AOM are trapped in the lysosomes and may therefore serve as lysosomal markers in subcellular fractionation studies. Vanadate reduced the amount of active surface asialoglycoprotein receptors approximately 70%, but had no effect on the rate of internalization and retroendocytosis of ligand. The amount of surface asialoglycoprotein receptors can be reduced by lowering the incubation temperature gradually from 37 to 15 degrees C; vanadate affected only the temperature--sensitive receptors. Vanadate inhibited degradation of 125I-TC-AOM 70-80%. Degradation was much more sensitive to vanadate than binding; half-maximal effects were seen at approximately 1 mM vanadate for binding and approximately 0.1 mM vanadate for degradation. By subcellular fractionation in sucrose and Nycodenz gradients, it was shown that vanadate completely prevented the transfer of 125I-TC-AOM from endosomes to lysosomes. Therefore, the inhibition of degradation by vanadate was indirect; in the presence of vanadate, ligand did not gain access to the lysosomes. The limited degradation in the presence of vanadate took place in a prelysosomal compartment. Vanadate did not affect cell viability and ATP content

  13. The roles of bulk and interfacial molecular orientations in determining the performance of organic bilayer solar cells

    KAUST Repository

    Ngongang Ndjawa, Guy O.

    2014-09-09

    Molecular orientation plays a significant role in determining the performance of small molecule solar cells. Key photovoltaic processes in these cells are strongly dependent on how the molecules are oriented in the active layer. We isolate contributions arising from the bulk molecular orientations vs. those from interfacial orientations in ZnPc/C60 bilayer systems and we probe these contributions by comparing device pairs in which only the bulk or the interface differ. By controlling the orientation in the bulk the current can be strongly modulated, whereas controlling the interfacial molecular orientation and degree of intermixing mediate the voltage.

  14. The roles of bulk and interfacial molecular orientations in determining the performance of organic bilayer solar cells

    KAUST Repository

    Ngongang Ndjawa, Guy O.; Graham, Kenneth R.; Conron, Sarah; Erwin, Patrick; Li, Ruipeng; Chou, Kang Wei; Burkhard, George; Krishnan Jagadamma, Lethy; Hoke, Eric T.; McGehee, Michael D.; Thompson, Mark E.; Amassian, Aram

    2014-01-01

    Molecular orientation plays a significant role in determining the performance of small molecule solar cells. Key photovoltaic processes in these cells are strongly dependent on how the molecules are oriented in the active layer. We isolate contributions arising from the bulk molecular orientations vs. those from interfacial orientations in ZnPc/C60 bilayer systems and we probe these contributions by comparing device pairs in which only the bulk or the interface differ. By controlling the orientation in the bulk the current can be strongly modulated, whereas controlling the interfacial molecular orientation and degree of intermixing mediate the voltage.

  15. Receptor-mediated endocytosis generates nanomechanical force reflective of ligand identity and cellular property.

    Science.gov (United States)

    Zhang, Xiao; Ren, Juan; Wang, Jingren; Li, Shixie; Zou, Qingze; Gao, Nan

    2018-08-01

    Whether environmental (thermal, chemical, and nutrient) signals generate quantifiable, nanoscale, mechanophysical changes in the cellular plasma membrane has not been well elucidated. Assessment of such mechanophysical properties of plasma membrane may shed lights on fundamental cellular process. Atomic force microscopic (AFM) measurement of the mechanical properties of live cells was hampered by the difficulty in accounting for the effects of the cantilever motion and the associated hydrodynamic force on the mechanical measurement. These challenges have been addressed in our recently developed control-based AFM nanomechanical measurement protocol, which enables a fast, noninvasive, broadband measurement of the real-time changes in plasma membrane elasticity in live cells. Here we show using this newly developed AFM platform that the plasma membrane of live mammalian cells exhibits a constant and quantifiable nanomechanical property, the membrane elasticity. This mechanical property sensitively changes in response to environmental factors, such as the thermal, chemical, and growth factor stimuli. We demonstrate that different chemical inhibitors of endocytosis elicit distinct changes in plasma membrane elastic modulus reflecting their specific molecular actions on the lipid configuration or the endocytic machinery. Interestingly, two different growth factors, EGF and Wnt3a, elicited distinct elastic force profiles revealed by AFM at the plasma membrane during receptor-mediated endocytosis. By applying this platform to genetically modified cells, we uncovered a previously unknown contribution of Cdc42, a key component of the cellular trafficking network, to EGF-stimulated endocytosis at plasma membrane. Together, this nanomechanical AFM study establishes an important foundation that is expandable and adaptable for investigation of cellular membrane evolution in response to various key extracellular signals. © 2017 Wiley Periodicals, Inc.

  16. WHAT GOVERNS THE BULK VELOCITY OF THE JET COMPONENTS IN ACTIVE GALACTIC NUCLEI?

    International Nuclear Information System (INIS)

    Chai Bo; Cao Xinwu; Gu Minfeng

    2012-01-01

    We use a sample of radio-loud active galactic nuclei (AGNs) with measured black hole masses to explore the jet formation mechanisms in these sources. Based on Königl's inhomogeneous jet model, the jet parameters, such as the bulk motion Lorentz factor, magnetic field strength, and electron density in the jet, can be estimated with the very long baseline interferometry and X-ray data.. We find a significant correlation between black hole mass and the bulk Lorentz factor of the jet components for this sample, while no significant correlation is present between the bulk Lorentz factor and the Eddington ratio. The massive black holes will be spun up through accretion, as the black holes acquire mass and angular momentum simultaneously through accretion. Recent investigation indeed suggested that most supermassive black holes in elliptical galaxies have on average higher spins than the black holes in spiral galaxies, where random, small accretion episodes (e.g., tidally disrupted stars, accretion of molecular clouds) might have played a more important role. If this is true, then the correlation between black hole mass and the bulk Lorentz factor of the jet components found in this work implies that the motion velocity of the jet components is probably governed by the black hole spin. No correlation is found between the magnetic field strength at 10R S (R S = 2GM/c 2 is the Schwarzschild radius) in the jets and the bulk Lorentz factor of the jet components for this sample. This is consistent with the black hole spin scenario, i.e., the faster moving jets are magnetically accelerated by the magnetic fields threading the horizon of more rapidly rotating black holes. The results imply that the Blandford-Znajek mechanism may dominate over the Blandford-Payne mechanism for the jet acceleration, at least in these radio-loud AGNs.

  17. Estimation of track registration efficiency in solution medium and study of gamma irradiation effects on the bulk-etch rate and the activation energy for bulk etching of CR-39 (DOP) Solid State Nuclear Track Detector

    International Nuclear Information System (INIS)

    Kalsi, P.C.

    2010-01-01

    The fission track registration efficiency of diethylene glycol bis allyl carbonate (dioctyl phthalate doped) (CR-39 (DOP)) solid state nuclear track detector (SSNTD) in solution medium (K wet ) has been experimentally determined and is found to be (9.7 ± 0.5).10 -4 cm. This is in good agreement with the values of other SSNTDs. The gamma irradiation effects in the dose range of 50.0-220.0 kGy on the bulk etch rate, V b and the activation energy for bulk etching, E of this solid state nuclear track detector (SSNTD) have also been studied. It is observed that the bulk etch rates increase and the activation energies for bulk etching decrease with the increase in gamma dose. These results have been explained on the basis of scission of the detector due to gamma irradiation

  18. Polarized localization and borate-dependent degradation of the Arabidopsis borate transporter BOR1 in tobacco BY-2 cells.

    Science.gov (United States)

    Yamauchi, Noboru; Gosho, Tadashi; Asatuma, Satoru; Toyooka, Kiminori; Fujiwara, Toru; Matsuoka, Ken

    2013-01-01

    In Arabidopsis the borate transporter BOR1, which is located in the plasma membrane, is degraded in the presence of excess boron by an endocytosis-mediated mechanism. A similar mechanism was suggested in rice as excess boron decreased rice borate transporter levels, although in this case whether the decrease was dependent on an increase in degradation or a decrease in protein synthesis was not elucidated. To address whether the borate-dependent degradation mechanism is conserved among plant cells, we analyzed the fate of GFP-tagged BOR1 (BOR1-GFP) in transformed tobacco BY-2 cells. Cells expressing BOR1-GFP displayed GFP fluorescence at the plasma membrane, especially at the membrane between two attached cells. The plasma membrane signal was abolished when cells were incubated in medium with a high concentration of borate (3 to 5 mM). This decrease in BOR1-GFP signal was mediated by a specific degradation of the protein after internalization by endocytosis from the plasma membrane. Pharmacological analysis indicated that the decrease in BOR1-GFP largely depends on the increase in degradation rate and that the degradation was mediated by a tyrosine-motif and the actin cytoskeleton. Tyr mutants of BOR1-GFP, which has been shown to inhibit borate-dependent degradation in Arabidopsis root cells, did not show borate-dependent endocytosis in tobacco BY-2 cells. These findings indicate that the borate-dependent degradation machinery of the borate transporter is conserved among plant species.

  19. Tyrosine phosphorylation of Eps15 is required for ligand-regulated, but not constitutive, endocytosis

    DEFF Research Database (Denmark)

    Confalonieri, S; Salcini, A E; Puri, C

    2000-01-01

    for endocytosis of the epidermal growth factor receptor (EGFR), the prototypical ligand-inducible receptor, but not of the transferrin receptor (TfR), the prototypical constitutively internalized receptor. Eps15, an endocytic protein that is tyrosine phosphorylated by EGFR, is a candidate for such a function....... Here, we show that tyrosine phosphorylation of Eps15 is necessary for internalization of the EGFR, but not of the TfR. We mapped Tyr 850 as the major in vivo tyrosine phosphorylation site of Eps15. A phosphorylation-negative mutant of Eps15 acted as a dominant negative on the internalization...... of the EGFR, but not of the TfR. A phosphopeptide, corresponding to the phosphorylated sequence of Eps15, inhibited EGFR endocytosis, suggesting that phosphotyrosine in Eps15 serves as a docking site for a phosphotyrosine binding protein. Thus, tyrosine phosphorylation of Eps15 represents the first molecular...

  20. Thermally activated reaction–diffusion-controlled chemical bulk reactions of gases and solids

    Directory of Open Access Journals (Sweden)

    S. Möller

    2015-01-01

    Full Text Available The chemical kinetics of the reaction of thin films with reactive gases is investigated. The removal of thin films using thermally activated solid–gas to gas reactions is a method to in-situ control deposition inventory in vacuum and plasma vessels. Significant scatter of experimental deposit removal rates at apparently similar conditions was observed in the past, highlighting the need for understanding the underlying processes. A model based on the presence of reactive gas in the films bulk and chemical kinetics is presented. The model describes the diffusion of reactive gas into the film and its chemical interaction with film constituents in the bulk using a stationary reaction–diffusion equation. This yields the reactive gas concentration and reaction rates. Diffusion and reaction rate limitations are depicted in parameter studies. Comparison with literature data on tokamak co-deposit removal results in good agreement of removal rates as a function of pressure, film thickness and temperature.

  1. Bulk Heterojunction Organic Solar Cell Area-Dependent Parameter Fluctuation

    Directory of Open Access Journals (Sweden)

    A. J. Trindade

    2017-01-01

    Full Text Available Organic solar cell efficiency is known to be active area dependent and is usually a problem in the upscale factor for market applications. In this work, a detailed study of organic photovoltaic devices with active layer based on poly(3-hexylthiophene (P3HT and 1-(3-methoxycarbonyl-propyl-1-phenyl-(6,6C61 (PCBM is made, evaluating the effect of the change on the active area from 10−2 to 4 cm4. The device structure was kept simple in order to allow the understanding of the physical effects involved. Device figures of merit were extracted from the equivalent circuit using a genetic-based algorithm, and their relationship with the active area was compared. It is observed that the efficiency drops significantly with the active area increase (as the fill factor while the parallel and series resistance, adjusted to the active area, seems to be relatively constant and increases linearly, respectively. The short circuit current and the generated photocurrent also drop significantly with the active area increase. The open circuit voltage does not show major changes. These results are discussed considering the main influences for the observed efficiency data. Particularly, as the basic circuit model seems to fail to explain the macroscopic results, the behavior can be related with the enlargement of defect interaction.

  2. Frequency-dependent complex modulus at the glass transition in Pd40Ni10Cu30P20 bulk amorphous alloys

    International Nuclear Information System (INIS)

    Lee, M.L.; Li, Y.; Feng, Y.P.; Carter, W.C.

    2003-01-01

    We report frequency-dependent measurements of the dynamic elastic modulus of a Pd 40 Cu 30 Ni 10 P 20 bulk amorphous phase near its glass transition temperature. The storage and loss moduli exhibit a structural relaxation similar to those observed by other characterization techniques. Parameters obtained by fitting to the Vogel-Fulcher-Tamman equation and the Kohlrausch-Williams-Watts model exhibit similar behaviors to those other methods

  3. The role of exciton ionization processes in bulk heterojunction organic photovoltaic cells

    Science.gov (United States)

    Zou, Yunlong; Holmes, Russell

    2015-03-01

    Dissociating photogenerated excitons into their constituent charges is essential for efficient photoconversion in organic semiconductors. Organic photovoltaics cells (OPV) widely adopt a heterojunction architecture where dissociation is facilitated by charge transfer at a donor-acceptor (D-A) interface. Interestingly, recent work on MoOx/C60 Schottky OPVs has demonstrated that excitons in C60 may also undergo bulk-ionization to generate photocurrent, driven by the built-in field at the MoOx/C60 interface. Here, we show that bulk-ionization processes also contribute to the photocurrent in bulk heterojunction (BHJ) OPVs with fullerene-rich compositions. The short-circuit current density (JSC) in a MoOx/C60 Schottky OPVs shows almost no dependence on temperature down to 80 K. This characteristic of bulk-ionization allows the use of temperature-dependent measurements of JSC to distinguish dissociation by bulk-ionization from charge transfer at a D-A interface. For BHJ OPVs constructed using the D-A pairing of boron subphthalocyanine chloride (SubPc)-C60, bulk-ionization is found to contribute >10% of the total photocurrent and >30% of the photocurrent from C60. We further find that fullerene-rich SubPc-C60 BHJ OPVs show a larger open-circuit voltage (VOC) than evenly mixed BHJs due to the presence of bulk-ionization. This talk will examine the dependence of JSC and VOC on the relative fraction of dissociation by charge transfer and bulk-ionization processes.

  4. Fate of bulk organic matter, nitrogen, and pharmaceutically active compounds in batch experiments simulating soil aquifer treatment (SAT) using primary effluent

    KAUST Repository

    Abel, Chol D T

    2013-06-30

    Reduction of bulk organic matter, nitrogen, and pharmaceutically active compounds from primary effluent during managed aquifer recharge was investigated using laboratory-scale batch reactors. Biologically stable batch reactors were spiked with different concentrations of sodium azide to inhibit biological activity and probe the effect of microbial activity on attenuation of various pollutants of concern. The experimental results obtained revealed that removal of dissolved organic carbon correlated with active microbial biomass. Furthermore, addition of 2 mM of sodium azide affected nitrite-oxidizing bacteria leading to accumulation of nitrite-nitrogen in the reactors while an ammonium-nitrogen reduction of 95.5 % was achieved. Removal efficiencies of the hydrophilic neutral compounds phenacetin, paracetamol, and caffeine were independent of the extent of the active microbial biomass and were >90 % in all reactors, whereas removal of pentoxifylline was dependent on the biological stability of the reactor. However, hydrophobic ionic compounds exhibited removal efficiency >80 % in batch reactors with the highest biological activity as evidenced by high concentration of adenosine triphosphate. © 2013 Springer Science+Business Media Dordrecht.

  5. Fate of bulk organic matter, nitrogen, and pharmaceutically active compounds in batch experiments simulating soil aquifer treatment (SAT) using primary effluent

    KAUST Repository

    Abel, Chol D T; Sharma, Saroj K.; Maeng, Sungkyu; Magic-Knezev, Aleksandra; Kennedy, Maria Dolores; Amy, Gary L.

    2013-01-01

    Reduction of bulk organic matter, nitrogen, and pharmaceutically active compounds from primary effluent during managed aquifer recharge was investigated using laboratory-scale batch reactors. Biologically stable batch reactors were spiked with different concentrations of sodium azide to inhibit biological activity and probe the effect of microbial activity on attenuation of various pollutants of concern. The experimental results obtained revealed that removal of dissolved organic carbon correlated with active microbial biomass. Furthermore, addition of 2 mM of sodium azide affected nitrite-oxidizing bacteria leading to accumulation of nitrite-nitrogen in the reactors while an ammonium-nitrogen reduction of 95.5 % was achieved. Removal efficiencies of the hydrophilic neutral compounds phenacetin, paracetamol, and caffeine were independent of the extent of the active microbial biomass and were >90 % in all reactors, whereas removal of pentoxifylline was dependent on the biological stability of the reactor. However, hydrophobic ionic compounds exhibited removal efficiency >80 % in batch reactors with the highest biological activity as evidenced by high concentration of adenosine triphosphate. © 2013 Springer Science+Business Media Dordrecht.

  6. Impact of CH3NH3PbI3-PCBM bulk heterojunction active layer on the photovoltaic performance of perovskite solar cells

    Science.gov (United States)

    Chaudhary, Dhirendra K.; Kumar, Pankaj; Kumar, Lokendra

    2017-10-01

    We report here the impact of CH3NH3PbI3-PCBM bulk heterojunction (BHJ) active layer on the photovoltaic performance of perovskite solar cells. The solar cells were prepared in normal architecture on FTO coated glass substrates with compact TiO2 (c-TiO2) layer on FTO as electron transport layer (ETL) and poly(3-hexylthiophene) (P3HT) as hole transport layer (HTL). For comparison, a few solar cells were also prepared in planar heterojunction structure using CH3NH3PbI3 only as the active layer. The bulk heterojunction CH3NH3PbI3-PCBM active layer exhibited very large crystalline grains of 2-3 μm compared to ∼150 nm only in CH3NH3PbI3 active layer. Larger grains in bulk-heterojunction solar cells resulted in enhanced power conversion efficiency (PCE) through enhancement in all the photovoltaic parameters compared to planar heterojunction solar cells. The bulk-heterojunction solar cells exhibited ∼9.25% PCE with short circuit current density (Jsc) of ∼18.649 mA/cm2, open circuit voltage (Voc) of 0.894 V and Fill Factor (FF) of 0.554. There was ∼36.9% enhancement in the PCE of bulk-heterojunction solar cells compared to that of planar heterojunction solar cells. The larger grains are formed as a result of incorporation on PCBM in the active layer.

  7. Internalization mechanisms of the epidermal growth factor receptor after activation with different ligands.

    Directory of Open Access Journals (Sweden)

    Lasse Henriksen

    Full Text Available The epidermal growth factor receptor (EGFR regulates normal growth and differentiation, but dysregulation of the receptor or one of the EGFR ligands is involved in the pathogenesis of many cancers. There are eight ligands for EGFR, however most of the research into trafficking of the receptor after ligand activation focuses on the effect of epidermal growth factor (EGF and transforming growth factor-α (TGF-α. For a long time it was believed that clathrin-mediated endocytosis was the major pathway for internalization of the receptor, but recent work suggests that different pathways exist. Here we show that clathrin ablation completely inhibits internalization of EGF- and TGF-α-stimulated receptor, however the inhibition of receptor internalization in cells treated with heparin-binding EGF-like growth factor (HB-EGF or betacellulin (BTC was only partial. In contrast, clathrin knockdown fully inhibits EGFR degradation after all ligands tested. Furthermore, inhibition of dynamin function blocked EGFR internalization after stimulation with all ligands. Knocking out a number of clathrin-independent dynamin-dependent pathways of internalization had no effect on the ligand-induced endocytosis of the EGFR. We suggest that EGF and TGF-α lead to EGFR endocytosis mainly via the clathrin-mediated pathway. Furthermore, we suggest that HB-EGF and BTC also lead to EGFR endocytosis via a clathrin-mediated pathway, but can additionally use an unidentified internalization pathway or better recruit the small amount of clathrin remaining after clathrin knockdown.

  8. Internalization Mechanisms of the Epidermal Growth Factor Receptor after Activation with Different Ligands

    Science.gov (United States)

    Henriksen, Lasse; Grandal, Michael Vibo; Knudsen, Stine Louise Jeppe; van Deurs, Bo; Grøvdal, Lene Melsæther

    2013-01-01

    The epidermal growth factor receptor (EGFR) regulates normal growth and differentiation, but dysregulation of the receptor or one of the EGFR ligands is involved in the pathogenesis of many cancers. There are eight ligands for EGFR, however most of the research into trafficking of the receptor after ligand activation focuses on the effect of epidermal growth factor (EGF) and transforming growth factor-α (TGF-α). For a long time it was believed that clathrin-mediated endocytosis was the major pathway for internalization of the receptor, but recent work suggests that different pathways exist. Here we show that clathrin ablation completely inhibits internalization of EGF- and TGF-α-stimulated receptor, however the inhibition of receptor internalization in cells treated with heparin-binding EGF-like growth factor (HB-EGF) or betacellulin (BTC) was only partial. In contrast, clathrin knockdown fully inhibits EGFR degradation after all ligands tested. Furthermore, inhibition of dynamin function blocked EGFR internalization after stimulation with all ligands. Knocking out a number of clathrin-independent dynamin-dependent pathways of internalization had no effect on the ligand-induced endocytosis of the EGFR. We suggest that EGF and TGF-α lead to EGFR endocytosis mainly via the clathrin-mediated pathway. Furthermore, we suggest that HB-EGF and BTC also lead to EGFR endocytosis via a clathrin-mediated pathway, but can additionally use an unidentified internalization pathway or better recruit the small amount of clathrin remaining after clathrin knockdown. PMID:23472148

  9. Mechanism of aldolase control of sorting nexin 9 function in endocytosis.

    Science.gov (United States)

    Rangarajan, Erumbi S; Park, HaJeung; Fortin, Emanuelle; Sygusch, Jurgen; Izard, Tina

    2010-04-16

    Sorting nexin 9 (SNX9) functions in a complex with the GTPase dynamin-2 at clathrin-coated pits, where it provokes fission of vesicles to complete endocytosis. Here the SNX9.dynamin-2 complex binds to clathrin and adapter protein complex 2 (AP-2) that line these pits, and this occurs through interactions of the low complexity domain (LC4) of SNX9 with AP-2. Intriguingly, localization of the SNX9.dynamin-2 complex to clathrin-coated pits is blocked by interactions with the abundant glycolytic enzyme aldolase, which also binds to the LC4 domain of SNX9. The crystal structure of the LC4 motif of human SNX9 in complex with aldolase explains the biochemistry and biology of this interaction, where SNX9 binds near the active site of aldolase via residues 165-171 that are also required for the interactions of SNX9 with AP-2. Accordingly, SNX9 binding to aldolase is structurally precluded by the binding of substrate to the active site. Interactions of SNX9 with aldolase are far more extensive and differ from those of the actin-nucleating factor WASP with aldolase, indicating considerable plasticity in mechanisms that direct the functions of the aldolase as a scaffold protein.

  10. Mechanism of Aldolase Control of Sorting Nexin 9 Function in Endocytosis*

    Science.gov (United States)

    Rangarajan, Erumbi S.; Park, HaJeung; Fortin, Emanuelle; Sygusch, Jurgen; Izard, Tina

    2010-01-01

    Sorting nexin 9 (SNX9) functions in a complex with the GTPase dynamin-2 at clathrin-coated pits, where it provokes fission of vesicles to complete endocytosis. Here the SNX9·dynamin-2 complex binds to clathrin and adapter protein complex 2 (AP-2) that line these pits, and this occurs through interactions of the low complexity domain (LC4) of SNX9 with AP-2. Intriguingly, localization of the SNX9·dynamin-2 complex to clathrin-coated pits is blocked by interactions with the abundant glycolytic enzyme aldolase, which also binds to the LC4 domain of SNX9. The crystal structure of the LC4 motif of human SNX9 in complex with aldolase explains the biochemistry and biology of this interaction, where SNX9 binds near the active site of aldolase via residues 165–171 that are also required for the interactions of SNX9 with AP-2. Accordingly, SNX9 binding to aldolase is structurally precluded by the binding of substrate to the active site. Interactions of SNX9 with aldolase are far more extensive and differ from those of the actin-nucleating factor WASP with aldolase, indicating considerable plasticity in mechanisms that direct the functions of the aldolase as a scaffold protein. PMID:20129922

  11. Mechanism of Aldolase Control of Sorting Nexin 9 Function in Endocytosis

    Energy Technology Data Exchange (ETDEWEB)

    Rangarajan, Erumbi S.; Park, HaJeung; Fortin, Emanuelle; Sygusch, Jurgen; Izard, Tina (Scripps); (Montreal)

    2010-11-03

    Sorting nexin 9 (SNX9) functions in a complex with the GTPase dynamin-2 at clathrin-coated pits, where it provokes fission of vesicles to complete endocytosis. Here the SNX9-dynamin-2 complex binds to clathrin and adapter protein complex 2 (AP-2) that line these pits, and this occurs through interactions of the low complexity domain (LC4) of SNX9 with AP-2. Intriguingly, localization of the SNX9-dynamin-2 complex to clathrin-coated pits is blocked by interactions with the abundant glycolytic enzyme aldolase, which also binds to the LC4 domain of SNX9. The crystal structure of the LC4 motif of human SNX9 in complex with aldolase explains the biochemistry and biology of this interaction, where SNX9 binds near the active site of aldolase via residues 165-171 that are also required for the interactions of SNX9 with AP-2. Accordingly, SNX9 binding to aldolase is structurally precluded by the binding of substrate to the active site. Interactions of SNX9 with aldolase are far more extensive and differ from those of the actin-nucleating factor WASP with aldolase, indicating considerable plasticity in mechanisms that direct the functions of the aldolase as a scaffold protein.

  12. Photocatalysis with chromium-doped TiO2: Bulk and surface doping

    KAUST Repository

    Ould-Chikh, Samy

    2014-04-15

    The photocatalytic properties of TiO2 modified by chromium are usually found to depend strongly on the preparation method. To clarify this problem, two series of chromium-doped titania with a chromium content of up to 1.56 wt % have been prepared under hydrothermal conditions: the first series (Cr:TiO2) is intended to dope the bulk of TiO2, whereas the second series (Cr/TiO2) is intended to load the surface of TiO2 with Cr. The catalytic properties have been compared in the photocatalytic oxidation of formic acid. Characterization data provides evidence that in the Cr/TiO2 catalysts chromium is located on the surface of TiO2 as amorphous CrOOH clusters. In contrast, in the Cr:TiO 2 series, chromium is mostly dissolved in the titania lattice, although a minor part is still present on the surface. Photocatalytic tests show that both series of chromium-doped titania demonstrate visible-light-driven photo-oxidation activity. Surface-doped Cr/TiO2 solids appear to be more efficient photocatalysts than the bulk-doped Cr:TiO2 counterparts. It\\'s classified! The photocatalytic properties of TiO2 modified by chromium depend strongly on the preparation method. To clarify this problem, two types of modified titania are discussed: one with CrIII doped in the bulk and one with CrOOH clusters on the TiO2 surface (see picture). Both series show visible-light-driven photo-oxidation activity. However, surface modification appears to be a more efficient strategy. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Mechanisms of the anomalous Pockels effect in bulk water

    Science.gov (United States)

    Yukita, Shunpei; Suzuki, Yuto; Shiokawa, Naoyuki; Kobayashi, Takayoshi; Tokunaga, Eiji

    2018-04-01

    The "anomalous" Pockels effect is a phenomenon that a light beam passing between two electrodes in an aqueous electrolyte solution is deflected by an AC voltage applied between the electrodes: the deflection angle is proportional to the voltage such that the incident beam alternately changes its direction. This phenomenon, the Pockels effect in bulk water, apparently contradicts what is believed in nonlinear optics, i.e., macroscopic inversion symmetry should be broken for the second-order nonlinear optical effect to occur such as the first-order electro-optic effect, i.e., the Pockels effect. To clarify the underlying mechanism, the dependence of the effect on the electrode material is investigated to find that the Pockels coefficient with Pt electrodes is two orders of magnitude smaller than with indium tin oxide (ITO) electrodes. It is experimentally confirmed that the Pockels effect of interfacial water in the electric double layer (EDL) on these electrodes shows an electrode dependence similar to the effect in bulk water while the effects depend on the frequency of the AC voltage such that the interfacial signal decreases with frequency but the bulk signal increases with frequency up to 221 Hz. These experimental results lead to a conclusion that the beam deflection is caused by the refractive index gradient in the bulk water region, which is formed transiently by the Pockels effect of interfacial water in the EDL when an AC electric field is applied. The refractive index gradient is caused by the diffuse layer spreading into the bulk region to work as a breaking factor of inversion symmetry of bulk water due to its charge-biased ionic distribution. This mechanism does not contradict the principle of nonlinear optics.

  14. The overexpressed human 46-kDa mannose 6-phosphate receptor mediates endocytosis and sorting of β-glucuronidase

    International Nuclear Information System (INIS)

    Watanabe, H.; Grubb, J.H.; Sly, W.S.

    1990-01-01

    The authors studied the function of the human small (46-kDa) mannose 6-phosphate receptor (SMPR) in transfected mouse L cells that do not express the larger insulin-like growth factor II/mannose 6-phosphate receptor. Cells overexpressing human SMPR were studied for enzyme binding to cell surface receptors, for binding to intracellular receptors in permeabilized cells, and for receptor-mediated endocytosis of recombinant human β-glucuronidase. Specific binding to human SMPR in permeabilized cells showed a pH optimum between pH 6.0 and pH 6.5. Binding was significant in the present of EDTA but was enhanced by added divalent cations. Up to 2.3% of the total functional receptor could be detected on the cell surface by enzyme binding. They present experiments showing that at very high levels of overexpression, and at pH 6.5, human SMPR mediated the endocytosis of β-glucuronidase. At pH 7.5, the rate of endocytosis was only 14% the rate seen at pH 6.5. Cells overexpressing human SMPR also showed reduced secretion of newly synthesized β-glucuronidase when compared to cells transfected with vector only, suggesting that overexpressed human SMPR can participate in sorting of newly synthesized β-glucuronidase and partially correct the sorting defect in mouse L cells that do not express the insulin-like growth factor II/mannose 6-phosphate receptor

  15. Construction and evaluation of BSA-CaP nanomaterials with enhanced transgene performance via biocorona-inspired caveolae-mediated endocytosis

    Science.gov (United States)

    Ma, Xi-Xi; Gao, Han; Zhang, Ya-Xuan; Jia, Yi-Yang; Li, Chen; Zhou, Si-Yuan; Zhang, Bang-Le

    2018-02-01

    Non-viral nanovectors have attracted much attention owing to their ability to condense genetic materials and their ease of modification. However, their poor stability, low biocompatibility and gene degradation in endosomes or lysosomes has significantly hampered their application in vivo and in the clinic. In an attempt to overcome these difficulties a series of bovine serum albumin (BSA)-calcium phosphate (CaP) nanoparticles were constructed. The CaP condenses with DNA to form nanocomplexes coated with a biomimetic corona of BSA. Such complexes may retain the inherent endocytosis profile of BSA, with improved biocompatibility. In particular the transgene performance may be enhanced by stimulating the cellular uptake pathway via caveolae-mediated endocytosis. Two methods were employed to construct and optimize the formulation of BSA-CaP nanomaterials. The optimized BSA-CaP-50-M2 nanoparticles prepared by our second method exhibited good stability, negligible cytotoxicity and enhanced transgene performance with long-term expression for 72 h in vivo even with a single dose. Determination of the cellular uptake pathway and Western blot revealed that cellular uptake of the designed BSA-CaP-50-M2 nanoparticles was mainly via caveolae-mediated endocytosis in a non-degradative pathway in which the biomimetic uptake profile of BSA was retained.

  16. BACE1 Is Necessary for Experience-Dependent Homeostatic Synaptic Plasticity in Visual Cortex

    Directory of Open Access Journals (Sweden)

    Emily Petrus

    2014-01-01

    Full Text Available Alzheimer’s disease (AD is the most common form of age-related dementia, which is thought to result from overproduction and/or reduced clearance of amyloid-beta (Aβ peptides. Studies over the past few decades suggest that Aβ is produced in an activity-dependent manner and has physiological relevance to normal brain functions. Similarly, physiological functions for β- and γ-secretases, the two key enzymes that produce Aβ by sequentially processing the amyloid precursor protein (APP, have been discovered over recent years. In particular, activity-dependent production of Aβ has been suggested to play a role in homeostatic regulation of excitatory synaptic function. There is accumulating evidence that activity-dependent immediate early gene Arc is an activity “sensor,” which acts upstream of Aβ production and triggers AMPA receptor endocytosis to homeostatically downregulate the strength of excitatory synaptic transmission. We previously reported that Arc is critical for sensory experience-dependent homeostatic reduction of excitatory synaptic transmission in the superficial layers of visual cortex. Here we demonstrate that mice lacking the major neuronal β-secretase, BACE1, exhibit a similar phenotype: stronger basal excitatory synaptic transmission and failure to adapt to changes in visual experience. Our results indicate that BACE1 plays an essential role in sensory experience-dependent homeostatic synaptic plasticity in the neocortex.

  17. Renormalization group approach to causal bulk viscous cosmological models

    International Nuclear Information System (INIS)

    Belinchon, J A; Harko, T; Mak, M K

    2002-01-01

    The renormalization group method is applied to the study of homogeneous and flat Friedmann-Robertson-Walker type universes, filled with a causal bulk viscous cosmological fluid. The starting point of the study is the consideration of the scaling properties of the gravitational field equations, the causal evolution equation of the bulk viscous pressure and the equations of state. The requirement of scale invariance imposes strong constraints on the temporal evolution of the bulk viscosity coefficient, temperature and relaxation time, thus leading to the possibility of obtaining the bulk viscosity coefficient-energy density dependence. For a cosmological model with bulk viscosity coefficient proportional to the Hubble parameter, we perform the analysis of the renormalization group flow around the scale-invariant fixed point, thereby obtaining the long-time behaviour of the scale factor

  18. Mechanisms of ceramide-induced COX-2-dependent apoptosis in human ovarian cancer OVCAR-3 cells partially overlapped with resveratrol

    DEFF Research Database (Denmark)

    Lin, Hung-Yun; Delmas, Dominique; Vang, Ole

    2013-01-01

    -2 appears at the apex of the p38 kinase-mediated signaling cascade induced by ceramide. Induction of apoptosis by ceramide or resveratrol was inhibited by the endocytosis inhibitor, cytochalasin D (CytD); however, cells exposed to resveratrol showed greater sensitivity than ceramide-treated cells....... Ceramide-treated cells underwent a dose-dependent reduction in trans-membrane potential. Although both ceramide and resveratrol induced the expressions of caspase-3 and -7, the effect of inducible COX-2 was different in caspase-7 expression induced by ceramide compared to resveratrol. In summary......, resveratrol and ceramide converge on an endocytosis-requiring, ERK1/2-dependent signal transduction pathway and induction of COX-expression as an essential molecular antecedent for subsequent p53-dependent apoptosis. In addition, expressions of caspase-3 and -7 are observed. However, a p38 kinase...

  19. Bulk-edge correspondence in topological transport and pumping

    Science.gov (United States)

    Imura, Ken-Ichiro; Yoshimura, Yukinori; Fukui, Takahiro; Hatsugai, Yasuhiro

    2018-03-01

    The bulk-edge correspondence (BEC) refers to a one-to-one relation between the bulk and edge properties ubiquitous in topologically nontrivial systems. Depending on the setup, BEC manifests in different forms and govern the spectral and transport properties of topological insulators and semimetals. Although the topological pump is theoretically old, BEC in the pump has been established just recently [1] motivated by the state-of-the-art experiments using cold atoms [2, 3]. The center of mass (CM) of a system with boundaries shows a sequence of quantized jumps in the adiabatic limit associated with the edge states. Despite that the bulk is adiabatic, the edge is inevitably non-adiabatic in the experimental setup or in any numerical simulations. Still the pumped charge is quantized and carried by the bulk. Its quantization is guaranteed by a compensation between the bulk and edges. We show that in the presence of disorder the pumped charge continues to be quantized despite the appearance of non-quantized jumps.

  20. Unlocking the Electrocatalytic Activity of Antimony for CO2 Reduction by Two-Dimensional Engineering of the Bulk Material.

    Science.gov (United States)

    Li, Fengwang; Xue, Mianqi; Li, Jiezhen; Ma, Xinlei; Chen, Lu; Zhang, Xueji; MacFarlane, Douglas R; Zhang, Jie

    2017-11-13

    Two-dimensional (2D) materials are known to be useful in catalysis. Engineering 3D bulk materials into the 2D form can enhance the exposure of the active edge sites, which are believed to be the origin of the high catalytic activity. Reported herein is the production of 2D "few-layer" antimony (Sb) nanosheets by cathodic exfoliation. Application of this 2D engineering method turns Sb, an inactive material for CO 2 reduction in its bulk form, into an active 2D electrocatalyst for reduction of CO 2 to formate with high efficiency. The high activity is attributed to the exposure of a large number of catalytically active edge sites. Moreover, this cathodic exfoliation process can be coupled with the anodic exfoliation of graphite in a single-compartment cell for in situ production of a few-layer Sb nanosheets and graphene composite. The observed increased activity of this composite is attributed to the strong electronic interaction between graphene and Sb. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Arf6-Dependent Intracellular Trafficking of Pasteurella multocida Toxin and pH-Dependent Translocation from Late Endosomes

    Directory of Open Access Journals (Sweden)

    Tracy P. M. Chong

    2011-03-01

    Full Text Available The potent mitogenic toxin from Pasteurella multocida (PMT is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn and cholera toxin (CT, the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

  2. Linking structure to fragility in bulk metallic glass-forming liquids

    International Nuclear Information System (INIS)

    Wei, Shuai; Stolpe, Moritz; Gross, Oliver; Gallino, Isabella; Hembree, William; Busch, Ralf; Evenson, Zach; Bednarcik, Jozef; Kruzic, Jamie J.

    2015-01-01

    Using in-situ synchrotron X-ray scattering, we show that the structural evolution of various bulk metallic glass-forming liquids can be quantitatively connected to their viscosity behavior in the supercooled liquid near T g . The structural signature of fragility is identified as the temperature dependence of local dilatation on distinct key atomic length scales. A more fragile behavior results from a more pronounced thermally induced dilatation of the structure on a length scale of about 3 to 4 atomic diameters, coupled with shallower temperature dependence of structural changes in the nearest neighbor environment. These findings shed light on the structural origin of viscous slowdown during undercooling of bulk metallic glass-forming liquids and demonstrate the promise of predicting the properties of bulk metallic glasses from the atomic scale structure

  3. Linking structure to fragility in bulk metallic glass-forming liquids

    Energy Technology Data Exchange (ETDEWEB)

    Wei, Shuai, E-mail: shuai.wei@asu.edu, E-mail: m.stolpe@mx.uni-saarland.de [Department of Materials Science and Engineering, Saarland University, Campus C63, 66123 Saarbrücken (Germany); Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287 (United States); Stolpe, Moritz, E-mail: shuai.wei@asu.edu, E-mail: m.stolpe@mx.uni-saarland.de; Gross, Oliver; Gallino, Isabella; Hembree, William; Busch, Ralf [Department of Materials Science and Engineering, Saarland University, Campus C63, 66123 Saarbrücken (Germany); Evenson, Zach [Department of Materials Science and Engineering, Saarland University, Campus C63, 66123 Saarbrücken (Germany); Institut für Materialphysik im Weltraum, Deutsches Zentrum für Luft- und Raumfahrt (DLR), 51170 Köln (Germany); Bednarcik, Jozef [Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, D-22603 Hamburg (Germany); Kruzic, Jamie J. [Material Science, School of Mechanical, Industrial, and Manufacturing Engineering, Oregon State University, Corvallis, Oregon 97331 (United States)

    2015-05-04

    Using in-situ synchrotron X-ray scattering, we show that the structural evolution of various bulk metallic glass-forming liquids can be quantitatively connected to their viscosity behavior in the supercooled liquid near T{sub g}. The structural signature of fragility is identified as the temperature dependence of local dilatation on distinct key atomic length scales. A more fragile behavior results from a more pronounced thermally induced dilatation of the structure on a length scale of about 3 to 4 atomic diameters, coupled with shallower temperature dependence of structural changes in the nearest neighbor environment. These findings shed light on the structural origin of viscous slowdown during undercooling of bulk metallic glass-forming liquids and demonstrate the promise of predicting the properties of bulk metallic glasses from the atomic scale structure.

  4. Probing endocytosis from the enterocyte brush border using fluorescent lipophilic dyes

    DEFF Research Database (Denmark)

    Danielsen, E Michael

    2015-01-01

    The small intestinal brush border is a specialized cell membrane that needs to withstand the solubilizing effect of bile salts during assimilation of dietary nutrients and to achieve detergent resistance; it is highly enriched in glycolipids organized in lipid raft microdomains. In the present work......-toluenesulfonate), and CellMask Orange plasma membrane stain were used to study endocytosis from the enterocyte brush border of organ-cultured porcine mucosal explants. All the dyes readily incorporated into the brush border but were not detectably endocytosed by 5 min, indicating a slow uptake compared with other cell types...

  5. Mechanism of Aldolase Control of Sorting Nexin 9 Function in Endocytosis*

    OpenAIRE

    Rangarajan, Erumbi S.; Park, HaJeung; Fortin, Emanuelle; Sygusch, Jurgen; Izard, Tina

    2010-01-01

    Sorting nexin 9 (SNX9) functions in a complex with the GTPase dynamin-2 at clathrin-coated pits, where it provokes fission of vesicles to complete endocytosis. Here the SNX9·dynamin-2 complex binds to clathrin and adapter protein complex 2 (AP-2) that line these pits, and this occurs through interactions of the low complexity domain (LC4) of SNX9 with AP-2. Intriguingly, localization of the SNX9·dynamin-2 complex to clathrin-coated pits is blocked by interactions with the abundant glycolytic ...

  6. A superconducting conveyer system using multiple bulk Y-Ba-Cu-O superconductors and permanent magnets

    Science.gov (United States)

    Kinoshita, T.; Koshizuka, N.; Nagashima, K.; Murakami, M.

    Developments of non-contact superconducting devices like superconducting magnetic levitation transfer and superconducting flywheel energy storage system have been performed based on the interactions between bulk Y-Ba-Cu-O superconductors and permanent magnets, in that the superconductors can stably be levitated without any active control. The performances of noncontact superconducting devices are dependent on the interaction forces like attractive forces and stiffness. In the present study, we constructed a non-contact conveyer for which the guide rails were prepared by attaching many Fe-Nd-B magnets onto an iron base plate. Along the translational direction, all the magnets were arranged as to face the same pole, and furthermore their inter-distance was made as small as possible. The guide rail has three magnet rows, for which the magnets were glued on the iron plate such that adjacent magnet rows have opposite poles like NSN. At the center row, the magnetic field at zero gap reached 0.61T, while the field strengths of two rows on the side edges were only 0.48T due to magnetic interactions among permanent magnets. We then prepared a cryogenic box made with FRP that can store several bulk Y-Ba-Cu-O superconductors 25 mm in diameter cooled by liquid nitrogen. It was found that the levitation forces and stiffness increased with increasing the number of bulk superconductors installed in the box, although the levitation force per unit bulk were almost the same. We also confirmed that these forces are dependent on the configuration of bulk superconductors.

  7. Analysis of equations of state and temperature dependence of thermal expansivity and bulk modulus for silicon

    International Nuclear Information System (INIS)

    Pandya, Tushar C; Bhatt, Apoorva D; Thakar, Nilesh A

    2012-01-01

    In the present paper an attempt has been made for the comparative study of different equations of state for silicon (Phase-1, cubic diamond structure) in the pressure range of 0-11 GPa. We compare the results of different equations of state (EOS) with available experimental data. The Kwon and Kim EOS is found to give far better agreement with the available experimental data. Results obtained by Poirier-Tarantola, Vinet, Tait and Suzuki's equations of state are not giving satisfactory agreement with the available experimental data. In the present study simple methods based on thermodynamic functions are presented to investigate the temperature dependence of thermal expansivity and bulk modulus for silicon. The results are reported for silicon. The calculated values of thermal expansivity are in good agreement with experimental data.

  8. Thermal, spectral, and surface properties of LED light-polymerized bulk fill resin composites.

    Science.gov (United States)

    Pişkin, Mehmet Burçin; Atalı, Pınar Yılmaz; Figen, Aysel Kantürk

    2015-02-01

    The aim of this study was to evaluate the thermal, spectral, and surface properties of four different bulk fill materials – SureFil SDR (SDR, Dentsplay DETREY), QuixFil (QF, Dentsplay DETREY), X-tra base (XB, Voco) X-tra fil (XF, Voco) – polymerized by light-emitting diode (LED). Resin matrix, filler type, size and amount, and photoinitiator types influence the degree of conversion. LED-cured bulk fill composites achieved sufficient polymerization. Scanning electron microscope (SEM) analysis revealed different patterns of surface roughness, depending on the composite material. Bulk fill materials showed surface characteristics similar to those of nanohybrid composites. Based on the thermal analysis results, glass transition (T(g)) and initial degradation (T(i)) temperatures changed depending on the bulk fill resin composites.

  9. Trapped magnetic field measurements on HTS bulk by peak controlled pulsed field magnetization

    International Nuclear Information System (INIS)

    Ida, Tetsuya; Watasaki, Masahiro; Kimura, Yosuke; Miki, Motohiro; Izumi, Mitsuru

    2010-01-01

    For the past several years, we have studied the high-temperature superconducting (HTS) synchronous motor assembled with melt-textured Gd-Ba-Cu-O bulk magnets. If the single pulse field magnetizes a bulk effectively, size of electrical motor will become small for the strong magnetic field of the HTS magnets without reducing output power of motor. In the previous study, we showed that the HTS bulk was magnetized to excellent cone-shape magnetic field distribution by using the waveform control pulse magnetization (WCPM) method. The WCPM technique made possible the active control of the waveform on which magnetic flux motion depended. We generated the pulse waveform with controlled risetime for HTS bulk magnetization to suppress the magnetic flux motion which decreases magnetization efficiency. The pulsed maximum magnetic flux density with slow risetime is not beyond the maximum magnetic flux density which is trapped by the static field magnetization. But, as for applying the pulse which has fast risetime, the magnetic flux which exceed greatly the threshold penetrates the bulk and causes the disorder of the trapped magnetic distribution. This fact suggests the possibility that the threshold at pulsed magnetization influences the dynamic magnetic flux motion. In this study, Gd-Ba-Cu-O bulk is magnetized by the controlled arbitrary trapezoidal shape pulse, of which the maximum magnetic flux density is controlled not to exceed the threshold. We will present the trapped magnetic characteristics and the technique to generate the controlled pulsed field.

  10. Energy-dependent proton damage in silicon

    Energy Technology Data Exchange (ETDEWEB)

    Donegani, Elena Maria

    2017-09-29

    Non Ionizing Energy Loss (NIEL) in the sensor bulk is a limiting factor for the lifetime of silicon detectors. In this work, the proton-energy dependent bulkdamage is studied in n- and p-type silicon pad diodes. The samples are thin (200 μm thick), and oxygen enriched (bulk material types: MCz, standard or deepdiffused FZ). Irradiations are performed with 23 MeV, 188 MeV and 23 GeV protons; the 1 MeV neutron equivalent fluence assumes selected values in the range [0.1,3].10{sup 14} cm{sup -2}. In reverse bias, Current-Voltage (IV) and Capacitance-Voltage (CV) measurements are performed to electrically characterise the samples; in forward bias, IV and CV measurements point out the transition from lifetime to relaxationlike semiconductor after irradiation. By means of Thermally Stimulated Current (TSC) measurements, 13 bulk defects have been found after proton irradiation. Firstly, TSC spectra are analysed to obtain defect concentrations after defect filling at the conventional temperature T{sub fill} =10 K. Secondly, temperature dependent capture coefficients of bulk defects are explained, according to the multi-phonon process, from the analysis of TSC measurements at higher filling temperatures (T{sub fill}<130 K). Thirdly, a new method based on the SRH statistics and accounting for cluster-induced shift in activation energy is proposed; it allows to fully characterise bulk defects (in terms of activation energy, concentration and majority capture cross-section) and to distinguish between point- and cluster-like defects. A correlation is noted between the leakage current and the concentration of three deep defects (namely the V{sub 2}, V{sub 3} and H(220K) defects), for all the investigated bulk materials and types, and after all the considered proton energies and fluences. At least five defects are found to be responsible for the space charge, with positive contributions from the E(30K) and B{sub i}O{sub i} defects, or negative contributions from three deep

  11. Energy-dependent proton damage in silicon

    International Nuclear Information System (INIS)

    Donegani, Elena Maria

    2017-01-01

    Non Ionizing Energy Loss (NIEL) in the sensor bulk is a limiting factor for the lifetime of silicon detectors. In this work, the proton-energy dependent bulkdamage is studied in n- and p-type silicon pad diodes. The samples are thin (200 μm thick), and oxygen enriched (bulk material types: MCz, standard or deepdiffused FZ). Irradiations are performed with 23 MeV, 188 MeV and 23 GeV protons; the 1 MeV neutron equivalent fluence assumes selected values in the range [0.1,3].10 14 cm -2 . In reverse bias, Current-Voltage (IV) and Capacitance-Voltage (CV) measurements are performed to electrically characterise the samples; in forward bias, IV and CV measurements point out the transition from lifetime to relaxationlike semiconductor after irradiation. By means of Thermally Stimulated Current (TSC) measurements, 13 bulk defects have been found after proton irradiation. Firstly, TSC spectra are analysed to obtain defect concentrations after defect filling at the conventional temperature T fill =10 K. Secondly, temperature dependent capture coefficients of bulk defects are explained, according to the multi-phonon process, from the analysis of TSC measurements at higher filling temperatures (T fill <130 K). Thirdly, a new method based on the SRH statistics and accounting for cluster-induced shift in activation energy is proposed; it allows to fully characterise bulk defects (in terms of activation energy, concentration and majority capture cross-section) and to distinguish between point- and cluster-like defects. A correlation is noted between the leakage current and the concentration of three deep defects (namely the V 2 , V 3 and H(220K) defects), for all the investigated bulk materials and types, and after all the considered proton energies and fluences. At least five defects are found to be responsible for the space charge, with positive contributions from the E(30K) and B i O i defects, or negative contributions from three deep acceptors H(116K), H(140K) and H(152K).

  12. Occurrence and fate of bulk organic matter and pharmaceutically active compounds in managed aquifer recharge: A review

    KAUST Repository

    Maeng, Sungkyu

    2011-05-01

    Managed aquifer recharge (MAR) is a natural water treatment process that induces surface water to flow in response to a hydraulic gradient through soil/sediment and into a vertical or horizontal well. It is a relatively cost-effective, robust and sustainable technology. Detailed characteristics of bulk organic matter and the occurrence and fate of pharmaceutically active compounds (PhACs) during MAR processes such as bank filtration (BF) and artificial recharge (AR) were reviewed. Understanding the fate of bulk organic matter during BF and AR is an essential step in determining pre- and/or post-treatment requirements. Analysis of organic matter characteristics using a suite of analytical tools suggests that there is a preferential removal of non-humic substances during MAR. Different classes of PhACs were found to behave differently during BF and AR. Antibiotics, non-steroidal anti-inflammatory drugs (NSAIDs), beta blockers, and steroid hormones generally exhibited good removal efficiencies, especially for compounds having hydrophobic-neutral characteristics. However, anticonvulsants showed a persistent behavior during soil passage. There were also some redox-dependent PhACs. For example, X-ray contrast agents measured, as adsorbable organic iodine (AOI), and sulfamethoxazole (an antibiotic) degraded more favorably under anoxic conditions compared to oxic conditions. Phenazone-type pharmaceuticals (NSAIDs) exhibited better removal under oxic conditions. The redox transition from oxic to anoxic conditions during soil passage can enhance the removal of PhACs that are sensitive to redox conditions. In general, BF and AR can be included in a multi-barrier treatment system for the removal of PhACs. © 2011.

  13. Occurrence and fate of bulk organic matter and pharmaceutically active compounds in managed aquifer recharge: a review.

    Science.gov (United States)

    Maeng, Sung Kyu; Sharma, Saroj K; Lekkerkerker-Teunissen, Karin; Amy, Gary L

    2011-05-01

    Managed aquifer recharge (MAR) is a natural water treatment process that induces surface water to flow in response to a hydraulic gradient through soil/sediment and into a vertical or horizontal well. It is a relatively cost-effective, robust and sustainable technology. Detailed characteristics of bulk organic matter and the occurrence and fate of pharmaceutically active compounds (PhACs) during MAR processes such as bank filtration (BF) and artificial recharge (AR) were reviewed. Understanding the fate of bulk organic matter during BF and AR is an essential step in determining pre- and/or post-treatment requirements. Analysis of organic matter characteristics using a suite of analytical tools suggests that there is a preferential removal of non-humic substances during MAR. Different classes of PhACs were found to behave differently during BF and AR. Antibiotics, non-steroidal anti-inflammatory drugs (NSAIDs), beta blockers, and steroid hormones generally exhibited good removal efficiencies, especially for compounds having hydrophobic-neutral characteristics. However, anticonvulsants showed a persistent behavior during soil passage. There were also some redox-dependent PhACs. For example, X-ray contrast agents measured, as adsorbable organic iodine (AOI), and sulfamethoxazole (an antibiotic) degraded more favorably under anoxic conditions compared to oxic conditions. Phenazone-type pharmaceuticals (NSAIDs) exhibited better removal under oxic conditions. The redox transition from oxic to anoxic conditions during soil passage can enhance the removal of PhACs that are sensitive to redox conditions. In general, BF and AR can be included in a multi-barrier treatment system for the removal of PhACs. Copyright © 2011. Published by Elsevier Ltd.

  14. Huntingtin-associated protein-1 (HAP1) regulates endocytosis and interacts with multiple trafficking-related proteins.

    Science.gov (United States)

    Mackenzie, Kimberly D; Lim, Yoon; Duffield, Michael D; Chataway, Timothy; Zhou, Xin-Fu; Keating, Damien J

    2017-07-01

    Huntingtin-associated protein 1 (HAP1) was initially identified as a binding partner of huntingtin, mutations in which underlie Huntington's disease. Subcellular localization and protein interaction data indicate that HAP1 may be important in vesicle trafficking, cell signalling and receptor internalization. In this study, a proteomics approach was used for the identification of novel HAP1-interacting partners to attempt to shed light on the physiological function of HAP1. Using affinity chromatography with HAP1-GST protein fragments bound to Sepharose columns, this study identified a number of trafficking-related proteins that bind to HAP1. Interestingly, many of the proteins that were identified by mass spectrometry have trafficking-related functions and include the clathrin light chain B and Sec23A, an ER to Golgi trafficking vesicle coat component. Using co-immunoprecipitation and GST-binding assays the association between HAP1 and clathrin light chain B has been validated in vitro. This study also finds that HAP1 co-localizes with clathrin light chain B. In line with a physiological function of the HAP1-clathrin interaction this study detected a dramatic reduction in vesicle retrieval and endocytosis in adrenal chromaffin cells. Furthermore, through examination of transferrin endocytosis in HAP1 -/- cortical neurons, this study has determined that HAP1 regulates neuronal endocytosis. In this study, the interaction between HAP1 and Sec23A was also validated through endogenous co-immunoprecipitation in rat brain homogenate. Through the identification of novel HAP1 binding partners, many of which have putative trafficking roles, this study provides us with new insights into the mechanisms underlying the important physiological function of HAP1 as an intracellular trafficking protein through its protein-protein interactions. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Bulk viscous cosmology in early Universe

    Indian Academy of Sciences (India)

    The effect of bulk viscosity on the early evolution of Universe for a spatially homogeneous and isotropic Robertson-Walker model is considered. Einstein's field equations are solved by using `gamma-law' equation of state = ( - 1)ρ, where the adiabatic parameter gamma () depends on the scale factor of the model.

  16. IRS-1 acts as an endocytic regulator of IGF-I receptor to facilitate sustained IGF signaling.

    Science.gov (United States)

    Yoneyama, Yosuke; Lanzerstorfer, Peter; Niwa, Hideaki; Umehara, Takashi; Shibano, Takashi; Yokoyama, Shigeyuki; Chida, Kazuhiro; Weghuber, Julian; Hakuno, Fumihiko; Takahashi, Shin-Ichiro

    2018-04-11

    Insulin-like growth factor-I receptor (IGF-IR) preferentially regulates the long-term IGF activities including growth and metabolism. Kinetics of ligand-dependent IGF-IR endocytosis determines how IGF induces such downstream signaling outputs. Here, we find that the insulin receptor substrate (IRS)-1 modulates how long ligand-activated IGF-IR remains at the cell surface before undergoing endocytosis in mammalian cells. IRS-1 interacts with the clathrin adaptor complex AP2. IRS-1, but not an AP2-binding-deficient mutant, delays AP2-mediated IGF-IR endocytosis after the ligand stimulation. Mechanistically, IRS-1 inhibits the recruitment of IGF-IR into clathrin-coated structures; for this reason, IGF-IR avoids rapid endocytosis and prolongs its activity on the cell surface. Accelerating IGF-IR endocytosis via IRS-1 depletion induces the shift from sustained to transient Akt activation and augments FoxO-mediated transcription. Our study establishes a new role for IRS-1 as an endocytic regulator of IGF-IR that ensures sustained IGF bioactivity, independent of its classic role as an adaptor in IGF-IR signaling. © 2018, Yoneyama et al.

  17. Bulk and shear viscosities of the gluon plasma in a quasiparticle description

    CERN Document Server

    Bluhm, M; Redlich, K

    2011-01-01

    Bulk and shear viscosities of deconfined gluonic matter are investigated within an effective kinetic theory by describing the strongly interacting medium phenomenologically in terms of quasiparticle excitations with medium-dependent self-energies. In this approach, local conservation of energy and momentum follows from a Boltzmann-Vlasov type kinetic equation and guarantees thermodynamic self-consistency. We show that the resulting transport coefficients reproduce the parametric dependencies on temperature and coupling obtained in perturbative QCD at large temperatures and small running coupling. The extrapolation into the non-perturbative regime results in a decreasing specific shear viscosity with decreasing temperature, exhibiting a minimum in the vicinity of the deconfinement transition temperature, while the specific bulk viscosity is sizeable in this region falling off rapidly with increasing temperature. The temperature dependence of specific bulk and shear viscosities found within this quasiparticle d...

  18. Phospho-dependent binding of the clathrin AP2 adaptor complex to GABAA receptors regulates the efficacy of inhibitory synaptic transmission.

    Science.gov (United States)

    Kittler, Josef T; Chen, Guojun; Honing, Stephan; Bogdanov, Yury; McAinsh, Kristina; Arancibia-Carcamo, I Lorena; Jovanovic, Jasmina N; Pangalos, Menelas N; Haucke, Volker; Yan, Zhen; Moss, Stephen J

    2005-10-11

    The efficacy of synaptic inhibition depends on the number of gamma-aminobutyric acid type A receptors (GABA(A)Rs) expressed on the cell surface of neurons. The clathrin adaptor protein 2 (AP2) complex is a critical regulator of GABA(A)R endocytosis and, hence, surface receptor number. Here, we identify a previously uncharacterized atypical AP2 binding motif conserved within the intracellular domains of all GABA(A)R beta subunit isoforms. This AP2 binding motif (KTHLRRRSSQLK in the beta3 subunit) incorporates the major sites of serine phosphorylation within receptor beta subunits, and phosphorylation within this site inhibits AP2 binding. Furthermore, by using surface plasmon resonance, we establish that a peptide (pepbeta3) corresponding to the AP2 binding motif in the GABA(A)R beta3 subunit binds to AP2 with high affinity only when dephosphorylated. Moreover, the pepbeta3 peptide, but not its phosphorylated equivalent (pepbeta3-phos), enhanced the amplitude of miniature inhibitory synaptic current and whole cell GABA(A)R current. These effects of pepbeta3 on GABA(A)R current were occluded by inhibitors of dynamin-dependent endocytosis supporting an action of pepbeta3 on GABA(A)R endocytosis. Therefore phospho-dependent regulation of AP2 binding to GABA(A)Rs provides a mechanism to specify receptor cell surface number and the efficacy of inhibitory synaptic transmission.

  19. Active moss biomonitoring of trace elements with Sphagnum girgensohnii moss bags in relation to atmospheric bulk deposition in Belgrade, Serbia

    International Nuclear Information System (INIS)

    Anicic, M.; Tasic, M.; Frontasyeva, M.V.; Tomasevic, M.; Rajsic, S.; Mijic, Z.; Popovic, A.

    2009-01-01

    Active biomonitoring with wet and dry moss bags was used to examine trace element atmospheric deposition in the urban area of Belgrade. The element accumulation capability of Sphagnum girgensohnii Russow was tested in relation to atmospheric bulk deposition. Moss bags were mounted for five 3-month periods (July 2005-October 2006) at three representative urban sites. For the same period monthly bulk atmospheric deposition samples were collected. The concentrations of Al, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, and Pb were determined by instrumental neutron activation analyses and atomic absorption spectrometry. Significant accumulation of most elements occurred in the exposed moss bags compared with the initial moss content. High correlations between the elements in moss and bulk deposits were found for V, Cu, As, and Ni. The enrichment factors of the elements for both types of monitor followed the same pattern at the corresponding sites. - Accumulated trace elements in the moss Sphagnum girgensohnii reflect atmospheric deposition

  20. Morphology-Dependent Trap Formation in High Performance Polymer Bulk Heterojunction Solar Cells

    KAUST Repository

    Beiley, Zach M.

    2011-06-28

    Bulk heterojunction solar cells (BHJs) based on poly[N-9″-hepta- decanyl-2,7-carbazole- alt -5,5-(4′,7′-di-2-thienyl-2′, 1′,3′-benzothiadiazole)] (PCDTBT) can have internal quantum efficiencies approaching 100% but require active layers that are too thin to absorb more than ∼70% of the above band gap light. When the active layer thickness is increased so that the cell absorbs more light, the fi ll factor and open circuit voltage decrease rapidly, so that the overall power conversion efficiency decreases. We fi nd that hole-traps in the polymer, which we characterize using space-charge limited current measurements, play an important role in the performance of PCDTBT-based BHJs and may limit the active layer thickness. Recombination due to carrier trapping is not often considered in BHJs because it is not believed to be a dominant loss mechanism in the "fruit-fl y" P3HT system. Furthermore, we show that in contrast to P3HT, PCDTBT has only weak short-range molecular order, and that annealing at temperatures above the glass transition decreases the order in the π-π stacking. The decrease in structural order is matched by the movement of hole-traps deeper into the band gap, so that thermal annealing worsens hole transport in the polymer and reduces the efficiency of PCDTBTbased BHJs. These fi ndings suggest that P3HT is not prototypical of the new class of high efficiency polymers, and that further improvement of BHJ efficiencies will necessitate the study of high efficiency polymers with low structural order. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Morphology-Dependent Trap Formation in High Performance Polymer Bulk Heterojunction Solar Cells

    KAUST Repository

    Beiley, Zach M.; Hoke, Eric T.; Noriega, Rodrigo; Dacuñ a, Javier; Burkhard, George F.; Bartelt, Jonathan A.; Salleo, Alberto; Toney, Michael F.; McGehee, Michael D.

    2011-01-01

    Bulk heterojunction solar cells (BHJs) based on poly[N-9″-hepta- decanyl-2,7-carbazole- alt -5,5-(4′,7′-di-2-thienyl-2′, 1′,3′-benzothiadiazole)] (PCDTBT) can have internal quantum efficiencies approaching 100% but require active layers that are too thin to absorb more than ∼70% of the above band gap light. When the active layer thickness is increased so that the cell absorbs more light, the fi ll factor and open circuit voltage decrease rapidly, so that the overall power conversion efficiency decreases. We fi nd that hole-traps in the polymer, which we characterize using space-charge limited current measurements, play an important role in the performance of PCDTBT-based BHJs and may limit the active layer thickness. Recombination due to carrier trapping is not often considered in BHJs because it is not believed to be a dominant loss mechanism in the "fruit-fl y" P3HT system. Furthermore, we show that in contrast to P3HT, PCDTBT has only weak short-range molecular order, and that annealing at temperatures above the glass transition decreases the order in the π-π stacking. The decrease in structural order is matched by the movement of hole-traps deeper into the band gap, so that thermal annealing worsens hole transport in the polymer and reduces the efficiency of PCDTBTbased BHJs. These fi ndings suggest that P3HT is not prototypical of the new class of high efficiency polymers, and that further improvement of BHJ efficiencies will necessitate the study of high efficiency polymers with low structural order. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Binding and Endocytosis of Bovine Hololactoferrin by the Parasite Entamoeba histolytica

    Directory of Open Access Journals (Sweden)

    Guillermo Ortíz-Estrada

    2015-01-01

    Full Text Available Entamoeba histolytica is a human parasite that requires iron (Fe for its metabolic function and virulence. Bovine lactoferrin (B-Lf and its peptides can be found in the digestive tract after dairy products are ingested. The aim of this study was to compare virulent trophozoites recently isolated from hamster liver abscesses with nonvirulent trophozoites maintained for more than 30 years in cultures in vitro regarding their interaction with iron-charged B-Lf (B-holo-Lf. We performed growth kinetics analyses of trophozoites in B-holo-Lf and throughout several consecutive transfers. The virulent parasites showed higher growth and tolerance to iron than nonvirulent parasites. Both amoeba variants specifically bound B-holo-Lf with a similar Kd. However, averages of 9.45 × 105 and 6.65 × 106 binding sites/cell were found for B-holo-Lf in nonvirulent and virulent amoebae, respectively. Virulent amoebae bound more efficiently to human and bovine holo-Lf, human holo-transferrin, and human and bovine hemoglobin than nonvirulent amoebae. Virulent amoebae showed two types of B-holo-Lf binding proteins. Although both amoebae endocytosed this glycoprotein through clathrin-coated vesicles, the virulent amoebae also endocytosed B-holo-Lf through a cholesterol-dependent mechanism. Both amoeba variants secreted cysteine proteases cleaving B-holo-Lf. These data demonstrate that the B-Lf endocytosis is more efficient in virulent amoebae.

  3. Binding and Endocytosis of Bovine Hololactoferrin by the Parasite Entamoeba histolytica.

    Science.gov (United States)

    Ortíz-Estrada, Guillermo; Calderón-Salinas, Víctor; Shibayama-Salas, Mineko; León-Sicairos, Nidia; de la Garza, Mireya

    2015-01-01

    Entamoeba histolytica is a human parasite that requires iron (Fe) for its metabolic function and virulence. Bovine lactoferrin (B-Lf) and its peptides can be found in the digestive tract after dairy products are ingested. The aim of this study was to compare virulent trophozoites recently isolated from hamster liver abscesses with nonvirulent trophozoites maintained for more than 30 years in cultures in vitro regarding their interaction with iron-charged B-Lf (B-holo-Lf). We performed growth kinetics analyses of trophozoites in B-holo-Lf and throughout several consecutive transfers. The virulent parasites showed higher growth and tolerance to iron than nonvirulent parasites. Both amoeba variants specifically bound B-holo-Lf with a similar K d . However, averages of 9.45 × 10(5) and 6.65 × 10(6) binding sites/cell were found for B-holo-Lf in nonvirulent and virulent amoebae, respectively. Virulent amoebae bound more efficiently to human and bovine holo-Lf, human holo-transferrin, and human and bovine hemoglobin than nonvirulent amoebae. Virulent amoebae showed two types of B-holo-Lf binding proteins. Although both amoebae endocytosed this glycoprotein through clathrin-coated vesicles, the virulent amoebae also endocytosed B-holo-Lf through a cholesterol-dependent mechanism. Both amoeba variants secreted cysteine proteases cleaving B-holo-Lf. These data demonstrate that the B-Lf endocytosis is more efficient in virulent amoebae.

  4. Unified bulk-boundary correspondence for band insulators

    Science.gov (United States)

    Rhim, Jun-Won; Bardarson, Jens H.; Slager, Robert-Jan

    2018-03-01

    The bulk-boundary correspondence, a topic of intensive research interest over the past decades, is one of the quintessential ideas in the physics of topological quantum matter. Nevertheless, it has not been proven in all generality and has in certain scenarios even been shown to fail, depending on the boundary profiles of the terminated system. Here, we introduce bulk numbers that capture the exact number of in-gap modes, without any such subtleties in one spatial dimension. Similarly, based on these 1D bulk numbers, we define a new 2D winding number, which we call the pole winding number, that specifies the number of robust metallic surface bands in the gap as well as their topological character. The underlying general methodology relies on a simple continuous extrapolation from the bulk to the boundary, while tracking the evolution of Green's function's poles in the vicinity of the bulk band edges. As a main result we find that all the obtained numbers can be applied to the known insulating phases in a unified manner regardless of the specific symmetries. Additionally, from a computational point of view, these numbers can be effectively evaluated without any gauge fixing problems. In particular, we directly apply our bulk-boundary correspondence construction to various systems, including 1D examples without a traditional bulk-boundary correspondence, and predict the existence of boundary modes on various experimentally studied graphene edges, such as open boundaries and grain boundaries. Finally, we sketch the 3D generalization of the pole winding number by in the context of topological insulators.

  5. Soil permittivity response to bulk electrical conductivity for selected soil water sensors

    Science.gov (United States)

    Bulk electrical conductivity can dominate the low frequency dielectric loss spectrum in soils, masking changes in the real permittivity and causing errors in estimated water content. We examined the dependence of measured apparent permittivity (Ka) on bulk electrical conductivity in contrasting soil...

  6. Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response.

    Science.gov (United States)

    Haniu, Hisao; Saito, Naoto; Matsuda, Yoshikazu; Tsukahara, Tamotsu; Maruyama, Kayo; Usui, Yuki; Aoki, Kaoru; Takanashi, Seiji; Kobayashi, Shinsuke; Nomura, Hiroki; Okamoto, Masanori; Shimizu, Masayuki; Kato, Hiroyuki

    2013-09-01

    We examined the cytotoxicity of multi-walled carbon nanotubes (MWCNTs) and the resulting cytokine secretion in BEAS-2B cells or normal human bronchial epithelial cells (HBEpCs) in two types of culture media (Ham's F12 containing 10% FBS [Ham's F12] and serum-free growth medium [SFGM]). Cellular uptake of MWCNT was observed by fluorescent microscopy and analyzed using flow cytometry. Moreover, we evaluated whether MWCNT uptake was suppressed by 2 types of endocytosis inhibitors. We found that BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM showed similar biological responses, but BEAS-2B cells cultured in SFGM did not internalize MWCNTs, and the 50% inhibitory concentration value, i.e., the cytotoxicity, was increased by more than 10-fold. MWCNT uptake was suppressed by a clathrin-mediated endocytosis inhibitor and a caveolae-mediated endocytosis inhibitor in BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM. In conclusion, we suggest that BEAS-2B cells cultured in a medium containing serum should be used for the safety evaluation of nanomaterials as a model of normal human bronchial epithelial cells. However, the culture medium composition may affect the proteins that are expressed on the cytoplasmic membrane, which may influence the biological response to MWCNTs. Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. Characterization and bulk properties of oxides

    International Nuclear Information System (INIS)

    Sonder, E.; Connolly, T.F.

    1979-06-01

    The bulk properties of oxides are divided into two classes, intrinsic properties which depend solely on the identity of the material, and extrinsic ones, which differ for different samples of the same compound. Sources of tabulated numerical values of intrinsic properties are given and modern developments in information storage and retrieval are discussed. Extrinsic properties are shown to depend on defects and trace impurities in the samples. Techniques of trace impurity analysis are discussed and realistic limits of detection and accuracies are given for routine analyses

  8. Cotransporting Ion is a Trigger for Cellular Endocytosis of Transporter-Targeting Nanoparticles: A Case Study of High-Efficiency SLC22A5 (OCTN2)-Mediated Carnitine-Conjugated Nanoparticles for Oral Delivery of Therapeutic Drugs.

    Science.gov (United States)

    Kou, Longfa; Yao, Qing; Sun, Mengchi; Wu, Chunnuan; Wang, Jia; Luo, Qiuhua; Wang, Gang; Du, Yuqian; Fu, Qiang; Wang, Jian; He, Zhonggui; Ganapathy, Vadivel; Sun, Jin

    2017-09-01

    OCTN2 (SLC22A5) is a Na + -coupled absorption transporter for l-carnitine in small intestine. This study tests the potential of this transporter for oral delivery of therapeutic drugs encapsulated in l-carnitine-conjugated poly(lactic-co-glycolic acid) (PLGA) nanoparticles (LC-PLGA NPs) and discloses the molecular mechanism for cellular endocytosis of transporter-targeting nanoparticles. Conjugation of l-carnitine to a surface of PLGA-NPs enhances the cellular uptake and intestinal absorption of encapsulated drug. In both cases, the uptake process is dependent on cotransporting ion Na + . Computational OCTN2 docking analysis shows that the presence of Na + is important for the formation of the energetically stable intermediate complex of transporter-Na + -LC-PLGA NPs, which is also the first step in cellular endocytosis of nanoparticles. The transporter-mediated intestinal absorption of LC-PLGA NPs occurs via endocytosis/transcytosis rather than via the traditional transmembrane transport. The portal blood versus the lymphatic route is evaluated by the plasma appearance of the drug in the control and lymph duct-ligated rats. Absorption via the lymphatic system is the predominant route in the oral delivery of the NPs. In summary, LC-PLGA NPs can effectively target OCTN2 on the enterocytes for enhancing oral delivery of drugs and the critical role of cotransporting ions should be noticed in designing transporter-targeting nanoparticles. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Collapse of Incoherent Light Beams in Inertial Bulk Kerr Media

    DEFF Research Database (Denmark)

    Bang, Ole; Edmundson, Darran; Królikowski, Wieslaw

    1999-01-01

    We use the coherent density function theory to show that partially coherent beams are unstable and may collapse in inertial bulk Kerr media. The threshold power for collapse, and its dependence on the degree of coherence, is found analytically and checked-numerically. The internal dynamics of the...... of the walk-off modes is illustrated for collapsing and diffracting partially coherent beams.......We use the coherent density function theory to show that partially coherent beams are unstable and may collapse in inertial bulk Kerr media. The threshold power for collapse, and its dependence on the degree of coherence, is found analytically and checked-numerically. The internal dynamics...

  10. The cell-based L-glutathione protection assays to study endocytosis and recycling of plasma membrane proteins.

    Science.gov (United States)

    Cihil, Kristine M; Swiatecka-Urban, Agnieszka

    2013-12-13

    Membrane trafficking involves transport of proteins from the plasma membrane to the cell interior (i.e. endocytosis) followed by trafficking to lysosomes for degradation or to the plasma membrane for recycling. The cell based L-glutathione protection assays can be used to study endocytosis and recycling of protein receptors, channels, transporters, and adhesion molecules localized at the cell surface. The endocytic assay requires labeling of cell surface proteins with a cell membrane impermeable biotin containing a disulfide bond and the N-hydroxysuccinimide (NHS) ester at 4 ºC - a temperature at which membrane trafficking does not occur. Endocytosis of biotinylated plasma membrane proteins is induced by incubation at 37 ºC. Next, the temperature is decreased again to 4 ºC to stop endocytic trafficking and the disulfide bond in biotin covalently attached to proteins that have remained at the plasma membrane is reduced with L-glutathione. At this point, only proteins that were endocytosed remain protected from L-glutathione and thus remain biotinylated. After cell lysis, biotinylated proteins are isolated with streptavidin agarose, eluted from agarose, and the biotinylated protein of interest is detected by western blotting. During the recycling assay, after biotinylation cells are incubated at 37 °C to load endocytic vesicles with biotinylated proteins and the disulfide bond in biotin covalently attached to proteins remaining at the plasma membrane is reduced with L-glutathione at 4 ºC as in the endocytic assay. Next, cells are incubated again at 37 °C to allow biotinylated proteins from endocytic vesicles to recycle to the plasma membrane. Cells are then incubated at 4 ºC, and the disulfide bond in biotin attached to proteins that recycled to the plasma membranes is reduced with L-glutathione. The biotinylated proteins protected from L-glutathione are those that did not recycle to the plasma membrane.

  11. Transport properties of high-temperature superconductors: Surface vs bulk effect

    International Nuclear Information System (INIS)

    Burlachkov, L.; Koshelev, A.E.; Vinokur, V.M.

    1996-01-01

    We investigate surface-related transport properties of high-temperature superconductors. We find the mean vortex velocity under applied transport current determined by the activation energies for vortex penetration and exit through the Bean-Livingston barrier. We determine the current distribution between the surfaces of superconductor and the field and current dependencies of the transport activation energies. For a three-dimensional superconductor the transport activation energy, U s 3D , is found to decrease with the external field, H, and transport current, J, as U s 3D ∝H -1/2 and U s 3D ∝J -1/2 , respectively. In the quasi-two-dimensional compounds, U s 2D decays logarithmically with field and current. The interplay between the surface and the bulk contributions to the transport properties, such as current-voltage characteristics, is discussed. copyright 1996 The American Physical Society

  12. Rab11b mediates melanin transfer between donor melanocytes and acceptor keratinocytes via coupled exo/endocytosis.

    Science.gov (United States)

    Tarafder, Abul K; Bolasco, Giulia; Correia, Maria S; Pereira, Francisco J C; Iannone, Lucio; Hume, Alistair N; Kirkpatrick, Niall; Picardo, Mauro; Torrisi, Maria R; Rodrigues, Inês P; Ramalho, José S; Futter, Clare E; Barral, Duarte C; Seabra, Miguel C

    2014-04-01

    The transfer of melanin from melanocytes to keratinocytes is a crucial process underlying maintenance of skin pigmentation and photoprotection against UV damage. Here, we present evidence supporting coupled exocytosis of the melanin core, or melanocore, by melanocytes and subsequent endocytosis by keratinocytes as a predominant mechanism of melanin transfer. Electron microscopy analysis of human skin samples revealed three lines of evidence supporting this: (1) the presence of melanocores in the extracellular space; (2) within keratinocytes, melanin was surrounded by a single membrane; and (3) this membrane lacked the melanosomal membrane protein tyrosinase-related protein 1 (TYRP1). Moreover, co-culture of melanocytes and keratinocytes suggests that melanin exocytosis is specifically induced by keratinocytes. Furthermore, depletion of Rab11b, but not Rab27a, caused a marked decrease in both keratinocyte-stimulated melanin exocytosis and transfer to keratinocytes. Thus, we propose that the predominant mechanism of melanin transfer is keratinocyte-induced exocytosis, mediated by Rab11b through remodeling of the melanosome membrane, followed by subsequent endocytosis by keratinocytes.

  13. Characteristics of bulk liquid undercooling and crystallization ...

    Indian Academy of Sciences (India)

    Characteristics of bulk liquid undercooling and crystallization behaviors ... cooling rate is fixed, the change of undercooling depends on the melt processing tem- ... solidification and a deep knowledge of undercooling of ... evolution, to obtain the information for the nucleation and ..... When cooling rate is fixed, the change.

  14. Should bulk cloudwater or fogwater samples obey Henry's law

    Energy Technology Data Exchange (ETDEWEB)

    Pandis, S.N.; Seinfeld, J.H. (Department of Chemical Engineering and Environmental Quality Laboratory, California Institute of Technology, Pasadena, CA (USA))

    1991-06-20

    In this work we prove that mixing of droplets with different {ital p}H that are individually in Henry's law equilibrium with the surrounding atmosphere always results in a bulk mixture that is supersaturated with weak acids like S(IV) and HCOOH and bases like NH{sub 3} with respect to the original atmosphere. The degree of supersaturation of the bulk liquid water sample for a particular species depends on its dissociation constant, on the initial {ital p}H of the bulk droplet mixture, and on the distribution of the {ital p}H and of the liquid water over the droplet spectrum. High supersaturations result only when the {ital p}H of the bulk droplet mixture exceeds the {ital p}K{sub {ital a}} of the species, in which {ital p}H range large {ital p}H differences among droplets of different sizes lead to large deviations from Henry's law for the bulk mixture. The deviation is shown to depend on the ratio of the arithmetic mean to the harmonic mean of the hydrogen ion concentrations of the droplets with the liquid water content used was weighting factor in the calculation of the means. The theory developed can explain observed discrepancies from Henry's law in atmospheric samples and also other observed phenomena like the reported increase of {ital p}H values of bulk aqueous samples during storage. {copyright} American Geophysical Union 1991

  15. Polarization Dependent Bulk-sensitive Valence Band Photoemission Spectroscopy and Density Functional Theory Calculations: Part I. 3d Transition Metals

    Science.gov (United States)

    Ueda, Shigenori; Hamada, Ikutaro

    2017-12-01

    The X-ray polarization dependent valence band HAXPES spectra of 3d transition metals (TMs) of Ti-Zn were measured to investigate the orbital resolved electronic structures by utilizing that the fact the photoionization cross-section of the atomic orbitals strongly depends on the experimental geometry. We have calculated the HAXPES spectra, which correspond to the cross-section weighted densities of states (CSW-DOSs), where the DOSs were obtained by the density functional theory calculations, and we have determined the relative photoionization cross-sections of the 4s and 4p orbitals to the 3d orbital in the 3d TMs. The experimentally obtained bulk-sensitive 3d and 4s DOSs were good agreement with the calculated DOSs in Ti, V, Cr, and Cu. In contrast, the deviations between the experimental and calculated 3d DOSs for Mn, Fe, Co, Ni were found, suggesting that the electron correlation plays an important role in the electronic structures for these materials.

  16. Polarized localization and borate-dependent degradation of the Arabidopsis borate transporter BOR1 in tobacco BY-2 cells [v1; ref status: indexed, http://f1000r.es/kv

    Directory of Open Access Journals (Sweden)

    Noboru Yamauchi

    2013-09-01

    Full Text Available In Arabidopsis the borate transporter BOR1, which is located in the plasma membrane, is degraded in the presence of excess boron by an endocytosis-mediated mechanism. A similar mechanism was suggested in rice as excess boron decreased rice borate transporter levels, although in this case whether the decrease was dependent on an increase in degradation or a decrease in protein synthesis was not elucidated. To address whether the borate-dependent degradation mechanism is conserved among plant cells, we analyzed the fate of GFP-tagged BOR1 (BOR1-GFP in transformed tobacco BY-2 cells. Cells expressing BOR1-GFP displayed GFP fluorescence at the plasma membrane, especially at the membrane between two attached cells. The plasma membrane signal was abolished when cells were incubated in medium with a high concentration of borate (3 to 5 mM. This decrease in BOR1-GFP signal was mediated by a specific degradation of the protein after internalization by endocytosis from the plasma membrane. Pharmacological analysis indicated that the decrease in BOR1-GFP largely depends on the increase in degradation rate and that the degradation was mediated by a tyrosine-motif and the actin cytoskeleton. Tyr mutants of BOR1-GFP, which has been shown to inhibit borate-dependent degradation in Arabidopsis root cells, did not show borate-dependent endocytosis in tobacco BY-2 cells. These findings indicate that the borate-dependent degradation machinery of the borate transporter is conserved among plant species.

  17. Discriminatory Role of Detergent-Resistant Membranes in the Dimerization and Endocytosis of Prostate-Specific Membrane Antigen

    Science.gov (United States)

    Schmidt, Sonja; Gericke, Birthe; Fracasso, Giulio; Ramarli, Dunia; Colombatti, Marco; Naim, Hassan Y.

    2013-01-01

    Prostate-specific membrane antigen (PSMA) is a type-II membrane glycoprotein that was initially identified in LNCaP cells. It is expressed at elevated levels in prostate cancer. In view of the correlation between the expression levels of PSMA and disease grade and stage, PSMA is considered to be one of the most promising biomarkers in the diagnosis and treatment of prostate cancer. In LNCaP cells PSMA undergoes internalization via clathrin-coated pits followed by accumulation in the endosomes. PSMA associates with different types of detergent-resistant membranes (DRMs) along the secretory pathway. Its mature form is mainly insoluble in Lubrol WX, but does not associate with Triton X-100-DRMs. To understand the mechanism of PSMA internalization we investigated its association during internalization with DRMs. For this purpose, internalization was induced by antibody cross-linking. We demonstrate at the biochemical and cell biological levels that: [i] exclusively homodimers of PSMA are associated with Lubrol WX-DRMs, [ii] antibody-induced cross-linking of PSMA molecules results in a time-dependent partitioning into another DRMs type, namely Triton X-100-DRMs, and [iii] concomitant with its association with Triton-X-100-DRMs internalization of PSMA occurs along tubulin filaments. In a previous work (Colombatti et al. (2009) PLoS One 4: e4608) we demonstrated that the small GTPases RAS and RAC1 and the MAPKs p38 and ERK1/2 are activated during antibody cross-linking. As downstream effects of this activation we observed a strong induction of NF-kB associated with an increased expression of IL-6 and CCL5 genes and that IL-6 and CCL5 enhanced the proliferative potential of LNCaP cells synergistically. These observations together with findings reported here hypothesize a fundamental role of DRMs during activation of PSMA as platforms for trafficking, endocytosis and signalling. Understanding these mechanisms constitutes an essential prerequisite for utilization of PSMA as

  18. Discriminatory Role of Detergent-Resistant Membranes in the Dimerization and Endocytosis of Prostate-Specific Membrane Antigen.

    Directory of Open Access Journals (Sweden)

    Sonja Schmidt

    Full Text Available Prostate-specific membrane antigen (PSMA is a type-II membrane glycoprotein that was initially identified in LNCaP cells. It is expressed at elevated levels in prostate cancer. In view of the correlation between the expression levels of PSMA and disease grade and stage, PSMA is considered to be one of the most promising biomarkers in the diagnosis and treatment of prostate cancer. In LNCaP cells PSMA undergoes internalization via clathrin-coated pits followed by accumulation in the endosomes. PSMA associates with different types of detergent-resistant membranes (DRMs along the secretory pathway. Its mature form is mainly insoluble in Lubrol WX, but does not associate with Triton X-100-DRMs. To understand the mechanism of PSMA internalization we investigated its association during internalization with DRMs. For this purpose, internalization was induced by antibody cross-linking. We demonstrate at the biochemical and cell biological levels that: [i] exclusively homodimers of PSMA are associated with Lubrol WX-DRMs, [ii] antibody-induced cross-linking of PSMA molecules results in a time-dependent partitioning into another DRMs type, namely Triton X-100-DRMs, and [iii] concomitant with its association with Triton-X-100-DRMs internalization of PSMA occurs along tubulin filaments. In a previous work (Colombatti et al. (2009 PLoS One 4: e4608 we demonstrated that the small GTPases RAS and RAC1 and the MAPKs p38 and ERK1/2 are activated during antibody cross-linking. As downstream effects of this activation we observed a strong induction of NF-kB associated with an increased expression of IL-6 and CCL5 genes and that IL-6 and CCL5 enhanced the proliferative potential of LNCaP cells synergistically. These observations together with findings reported here hypothesize a fundamental role of DRMs during activation of PSMA as platforms for trafficking, endocytosis and signalling. Understanding these mechanisms constitutes an essential prerequisite for

  19. Bulk Concentration Dependence of Electrolyte Resistance Within Mesopores of Carbon Electrodes in Electric Double-Layer Capacitors

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jaekwang; Kim, Daeun; Lee, Ilbok; Son, Hyungbin; Lee, Donghyun; Yoon, Songhun [Chung-Ang University, Seoul (Korea, Republic of); Shim, Hyewon [Korea Institute of Nuclear Nonproliferation and Control, Daejeon (Korea, Republic of); Lee, Jinwoo [POSTECH, Pohang (Korea, Republic of)

    2016-02-15

    Hexagonally ordered mesoporous carbon materials were prepared and used as electrode materials in an electric double-layer capacitor. Using this electrode, the change of electrolyte resistance within the mesopores was investigated according to the bulk electrolyte concentration. Using three different electrochemical transient experiments-imaginary capacitance analysis, chronoamperometry, and hronopotentiometry-the time constant associated with electrolyte transport was determined, which was then used to obtain the electrolyte resistance within the mesopores. With decreasing electrolyte concentration, the increase in electrolyte resistance was smaller than the increase in the resistivity of the bulk electrolyte, which is indicative of a different environment for ionic transport within the mesopores. On using the confinement effect within the mesopores, the predicted higher concentration within mesopore probably results in lower electrolyte resistance, especially under low bulk concentrations.

  20. The dependence of Pi2 waveforms on periodic velocity enhancements within bursty bulk flows

    Directory of Open Access Journals (Sweden)

    K. R. Murphy

    2011-03-01

    Full Text Available Pi2s are a category of Ultra Low Frequency (ULF waves associated with the onset of magnetic substorms. Recent work has suggested that the deceleration of bulk plasma flows in the central plasmasheet, known as bursty bulk flows (BBFs, are able to directly-drive Pi2 oscillations. Some of these studies have further shown evidence that there is a one-to-one correlation between Pi2 magnetic waveforms observed on the ground and periodic peaks in flow velocity within the BBF, known as flow bursts. Utilising a favourable conjunction between the Geotail spacecraft and the Canadian Array for Real-time Investigations of Magnetic Activity (CARISMA magnetometer array on 31 May 1998, we examine the causality of the link between BBF flow bursts and Pi2 waveforms. Using a series of analytical tests in both the time and frequency domains, we find that while the Pi2 and BBF waveforms are very similar, the ground response for this event occurs prior to the observed flow enhancements in the magnetotail. We conclude that during this specific case study the temporal variations of the flow bursts within the BBF are not directly-driving the observed ground-based Pi2 waveforms, despite the fact that a visual inspection of both time-series might initially suggest that there is a causal relationship. We postulate that rather than there being a direct causal relation, the similar waveforms observed in both Pi2s and BBFs may result from temporal variations in a common source for both the BBFs and the Pi2s, such as magnetic reconnection in the tail, this source modulating both the Pi2 and BBF at the same frequency.

  1. Enhanced infrared magneto-optical response of the nonmagnetic semiconductor BiTeI driven by bulk Rashba splitting

    Energy Technology Data Exchange (ETDEWEB)

    Demko, L.; Tokura, Y. [Multiferroics Project, ERATO, JST, c/o Department of Applied Physics, University of Tokyo (Japan); Schober, G.A.H. [Institute for Theoretical Physics, University of Heidelberg (Germany); Kocsis, V.; Kezsmarki, I. [Department of Physics, Budapest University of Technology and Economics and Condensed Matter Research Group of the Hungarian Academy of Sciences (Hungary); Bahramy, M.S.; Murakawa, H. [CMRG and CERG, RIKEN ASI (Japan); Lee, J.S.; Arita, R.; Nagaosa, N. [Department of Applied Physics, University of Tokyo (Japan)

    2013-07-01

    We study the magneto-optical (MO) response of the polar semiconducting BiTeI with giant bulk Rashba spin splitting at various carrier densities. Despite being nonmagnetic, the material is found to yield a huge MO activity in the infrared region under moderate magnetic fields (up to 3 T). Our first-principles calculations show that the enhanced MO response of BiTeI comes mainly from the intraband transitions between the Rashba-split bulk conduction bands. These transitions connecting electronic states with opposite spin directions become active due to the presence of strong spin-orbit interaction and give rise to distinct features in the MO spectra with a systematic doping dependence. We predict an even more pronounced enhancement in the low-energy MO response and dc Hall effect near the crossing (Dirac) point of the conduction bands.

  2. Proper design of silica nanoparticles combines high brightness, lack of cytotoxicity and efficient cell endocytosis

    Science.gov (United States)

    Rampazzo, Enrico; Voltan, Rebecca; Petrizza, Luca; Zaccheroni, Nelsi; Prodi, Luca; Casciano, Fabio; Zauli, Giorgio; Secchiero, Paola

    2013-08-01

    Silica-based luminescent nanoparticles (SiNPs) show promising prospects in nanomedicine in light of their chemical properties and versatility. In this study, we have characterized silica core-PEG shell SiNPs derivatized with PEG moieties (NP-PEG), with external amino- (NP-PEG-amino) or carboxy-groups (NP-PEG-carbo), both in cell cultures as well as in animal models. By using different techniques, we could demonstrate that these SiNPs were safe and did not exhibit appreciable cytotoxicity in different relevant cell models, of normal or cancer cell types, growing either in suspension (JVM-2 leukemic cell line and primary normal peripheral blood mononuclear cells) or in adherence (human hepatocarcinoma Huh7 and umbilical vein endothelial cells). Moreover, by multiparametric flow cytometry, we could demonstrate that the highest efficiency of cell uptake and entry was observed with NP-PEG-amino, with a stable persistence of the fluorescence signal associated with SiNPs in the loaded cell populations both in vitro and in vivo settings suggesting this as an innovative method for cell traceability and detection in whole organisms. Finally, experiments performed with the endocytosis inhibitor Genistein clearly suggested the involvement of a caveolae-mediated pathway in SiNP endocytosis. Overall, these data support the safe use of these SiNPs for diagnostic and therapeutic applications.Silica-based luminescent nanoparticles (SiNPs) show promising prospects in nanomedicine in light of their chemical properties and versatility. In this study, we have characterized silica core-PEG shell SiNPs derivatized with PEG moieties (NP-PEG), with external amino- (NP-PEG-amino) or carboxy-groups (NP-PEG-carbo), both in cell cultures as well as in animal models. By using different techniques, we could demonstrate that these SiNPs were safe and did not exhibit appreciable cytotoxicity in different relevant cell models, of normal or cancer cell types, growing either in suspension (JVM-2

  3. The carboxyl terminus of human cytomegalovirus-encoded 7 transmembrane receptor US28 camouflages agonism by mediating constitutive endocytosis

    DEFF Research Database (Denmark)

    Waldhoer, Maria; Casarosa, Paola; Rosenkilde, Mette M

    2003-01-01

    are separable entities in this viral chemokine receptor. We generated chimeric and mutant US28 proteins that were altered in either their constitutive endocytic (US28 Delta 300, US28 Delta 317, US28-NK1-ctail, and US28-ORF74-ctail) or signaling properties (US28R129A). By using this series of mutants, we show...... further show that the constitutive endocytic property of US28 affects the action of its chemokine ligand fractalkine/CX3CL1 and show that in the absence of the US28 C terminus, fractalkine/CX3CL1 acts as an agonist on US28. This demonstrates for the first time that the endocytic properties of a 7TM......US28 is one of four 7 transmembrane (7TM) chemokine receptors encoded by human cytomegalovirus and has been shown to both signal and endocytose in a ligand-independent, constitutively active manner. Here we show that the constitutive activity and constitutive endocytosis properties of US28...

  4. Visualization of the endocytic pathway in the filamentous fungus Aspergillus oryzae using an EGFP-fused plasma membrane protein

    International Nuclear Information System (INIS)

    Higuchi, Yujiro; Nakahama, Tomoyuki; Shoji, Jun-ya; Arioka, Manabu; Kitamoto, Katsuhiko

    2006-01-01

    Endocytosis is an important process for cellular activities. However, in filamentous fungi, the existence of endocytosis has been so far elusive. In this study, we used AoUapC-EGFP, the fusion protein of a putative uric acid-xanthine permease with enhanced green fluorescent protein (EGFP) in Aspergillus oryzae, to examine whether the endocytic process occurs or not. Upon the addition of ammonium into the medium the fusion protein was internalized from the plasma membrane. The internalization of AoUapC-EGFP was completely blocked by sodium azide, cold, and cytochalasin A treatments, suggesting that the internalization possesses the general features of endocytosis. These results demonstrate the occurrence of endocytosis in filamentous fungi. Moreover, we discovered that the endosomal compartments appeared upon the induction of endocytosis and moved in a microtubule-dependent manner

  5. Effects of bulk charged impurities on the bulk and surface transport in three-dimensional topological insulators

    Energy Technology Data Exchange (ETDEWEB)

    Skinner, B.; Chen, T.; Shklovskii, B. I., E-mail: shklovsk@physics.spa.umn.edu [University of Minnesota, Fine Theoretical Physics Institute (United States)

    2013-09-15

    In the three-dimensional topological insulator (TI), the physics of doped semiconductors exists literally side-by-side with the physics of ultrarelativistic Dirac fermions. This unusual pairing creates a novel playground for studying the interplay between disorder and electronic transport. In this mini-review, we focus on the disorder caused by the three-dimensionally distributed charged impurities that are ubiquitous in TIs, and we outline the effects it has on both the bulk and surface transport in TIs. We present self-consistent theories for Coulomb screening both in the bulk and at the surface, discuss the magnitude of the disorder potential in each case, and present results for the conductivity. In the bulk, where the band gap leads to thermally activated transport, we show how disorder leads to a smaller-than-expected activation energy that gives way to variable-range hopping at low temperatures. We confirm this enhanced conductivity with numerical simulations that also allow us to explore different degrees of impurity compensation. For the surface, where the TI has gapless Dirac modes, we present a theory of disorder and screening of deep impurities, and we calculate the corresponding zero-temperature conductivity. We also comment on the growth of the disorder potential in passing from the surface of the TI into the bulk. Finally, we discuss how the presence of a gap at the Dirac point, introduced by some source of time-reversal symmetry breaking, affects the disorder potential at the surface and the mid-gap density of states.

  6. Effects of bulk charged impurities on the bulk and surface transport in three-dimensional topological insulators

    International Nuclear Information System (INIS)

    Skinner, B.; Chen, T.; Shklovskii, B. I.

    2013-01-01

    In the three-dimensional topological insulator (TI), the physics of doped semiconductors exists literally side-by-side with the physics of ultrarelativistic Dirac fermions. This unusual pairing creates a novel playground for studying the interplay between disorder and electronic transport. In this mini-review, we focus on the disorder caused by the three-dimensionally distributed charged impurities that are ubiquitous in TIs, and we outline the effects it has on both the bulk and surface transport in TIs. We present self-consistent theories for Coulomb screening both in the bulk and at the surface, discuss the magnitude of the disorder potential in each case, and present results for the conductivity. In the bulk, where the band gap leads to thermally activated transport, we show how disorder leads to a smaller-than-expected activation energy that gives way to variable-range hopping at low temperatures. We confirm this enhanced conductivity with numerical simulations that also allow us to explore different degrees of impurity compensation. For the surface, where the TI has gapless Dirac modes, we present a theory of disorder and screening of deep impurities, and we calculate the corresponding zero-temperature conductivity. We also comment on the growth of the disorder potential in passing from the surface of the TI into the bulk. Finally, we discuss how the presence of a gap at the Dirac point, introduced by some source of time-reversal symmetry breaking, affects the disorder potential at the surface and the mid-gap density of states

  7. Should bulk cloudwater or fogwater samples obey Henry's law?

    Science.gov (United States)

    Pandis, Spyros N.; Seinfeld, John H.

    1991-06-01

    Mixing of droplets with different pH that are individually in Henry's law equilibrium with the surrounding atmosphere always results in a bulk mixture that is supersaturated with weak acids like S(IV) and HCOOH, and bases like NH3 with respect to the original atmosphere. High supersaturations result only when the pH of the bulk droplet mixture exceeds the pKa of the species, in which pH range large pH differences among droplets of different sizes lead to large deviations from Henry's law for the bulk mixture. The deviation is shown to depend on the ratio of the arithmetic mean to the harmonic mean of the hydrogen ion concentrations of the droplets with the liquid water content used as weighting factor in the calculation of the means. The theory developed can explain observed discrepancies from Henry's law in atmospheric samples and also other observed phenomena like the reported increase of pH values of bulk aqueous samples during storage.

  8. Impedance Spectroscopy and AC Conductivity Studies of Bulk 3-Amino-7-(dimethylamino)-2-methyl-hydrochloride

    Science.gov (United States)

    El-Shabaan, M. M.

    2018-02-01

    Impedance spectroscopy and alternating-current (AC) conductivity (σ AC) studies of bulk 3-amino-7-(dimethylamino)-2-methyl-hydrochloride (neutral red, NR) have been carried out over the temperature (T) range from 303 K to 383 K and frequency (f) range from 0.5 kHz to 5 MHz. Dielectric data were analyzed using the complex impedance (Z *) and complex electric modulus (M *) for bulk NR at various temperatures. The impedance loss peaks were found to shift towards high frequencies, indicating an increase in the relaxation time (τ 0) and loss in the material, with increasing temperature. For each temperature, a single depressed semicircle was observed at high frequencies, originating from the bulk transport, and a spike in the low-frequency region, resulting from the electrode effect. Fitting of these curves yielded an equivalent circuit containing a parallel combination of a resistance R and constant-phase element (CPE) Q. The carrier transport in bulk NR is governed by the correlated barrier hopping (CBH) mechanism, some parameters of which, such as the maximum barrier height (W M), charge density (N), and hopping distance (r), were determined as functions of both temperature and frequency. The frequency dependence of σ AC at different temperatures indicated that the conduction in bulk NR is a thermally activated process. The σ AC value at different frequencies increased linearly with temperature.

  9. Bulk energy storage increases United States electricity system emissions.

    Science.gov (United States)

    Hittinger, Eric S; Azevedo, Inês M L

    2015-03-03

    Bulk energy storage is generally considered an important contributor for the transition toward a more flexible and sustainable electricity system. Although economically valuable, storage is not fundamentally a "green" technology, leading to reductions in emissions. We model the economic and emissions effects of bulk energy storage providing an energy arbitrage service. We calculate the profits under two scenarios (perfect and imperfect information about future electricity prices), and estimate the effect of bulk storage on net emissions of CO2, SO2, and NOx for 20 eGRID subregions in the United States. We find that net system CO2 emissions resulting from storage operation are nontrivial when compared to the emissions from electricity generation, ranging from 104 to 407 kg/MWh of delivered energy depending on location, storage operation mode, and assumptions regarding carbon intensity. Net NOx emissions range from -0.16 (i.e., producing net savings) to 0.49 kg/MWh, and are generally small when compared to average generation-related emissions. Net SO2 emissions from storage operation range from -0.01 to 1.7 kg/MWh, depending on location and storage operation mode.

  10. Ac conductivity and dielectric properties of bulk tin phthalocyanine dichloride (SnPcCl 2)

    Science.gov (United States)

    El-Nahass, M. M.; Farid, A. M.; Abd El-Rahman, K. F.; Ali, H. A. M.

    2008-07-01

    The ac conductivity, σac( ω), has been measured for bulk tin phthalocyanine dichloride (SnPcCl 2) in the form of compressed pellet with evaporated ohmic Au electrodes in a temperature range 303-403 K. Ac conductivity, σac( ω), is found to vary as ωs in the frequency range 42 Hz-5×10 6 Hz. At low range of frequency, s<1 and it decreases with the increase in temperature indicating a dominant hopping process. At high range of frequency, s is found to be equal to ≈1.09 and is temperature independent. The dielectric constant, ε1, and dialectic loss, ε2, have been determined for bulk SnPcCl 2. Both ε1 and ε2 decrease with the increase in frequency and increase with the increase in temperature. The Cole-Cole types have been used to determine some parameters such as; the macroscopic relaxation time ( τo), the molecular relaxation time ( τ), the activation energy for relaxation ( Eo) and the distribution parameter ( α). The temperature dependence of τ is expressed by a thermally activated process with the activation energy of 0.299 eV.

  11. Pilot Control of Viscous Bulking in the Activated Sludge Treatment of Industrial Effluent from Soft Drink Plants

    Directory of Open Access Journals (Sweden)

    Mohammad Mehdi Esfahani

    2007-06-01

    Full Text Available Viscous bulking is a typical problem arising in activated sludge facilities treating effluent from soft drink plants. The drawbacks associated with this phenomenon include increased effluent organic loading and undesirable sludge settlement. In order to investigate this phenomenon, a soft drink factory was selected as a pilot plant for a case study (where metal tanks were used as a biological selector, an aeration basin, and a clarifier. The study shows that the major causes of viscous bulking are high organic loading and undesirable ratio of monovalent to divalent cations. In the biological selector (with a retention time of 20 hours, while the organic load in the influent to the aeration basin decreased by about 50%, with an impact on reduced viscous bulking, pH value decreased from 12 to 6.5 due to fatty acids production. Adjustment of Na/Ca ratio improved bacterial surface hydrophobicity and prevented degradation of biological flocs. This resulted in improved sludge settleability. Application of this method improved sludge settling, made flocs stronger, and reduced effluent organic load (COD to less than 150 mg/l, indicating stability of the system.

  12. Effects of exposure to nano and bulk sized TiO2 and CuO in Lemna minor.

    Science.gov (United States)

    Dolenc Koce, Jasna

    2017-10-01

    Nanoparticles of TiO 2 and CuO are among most commonly used nanoparticles, and elevated concentrations of them are expected to be found in all environments, including aquatic. A standard growth inhibition test ISO/CD 20079 was used to determine the toxicity of nano sized and larger micro sized (bulk) particles in the concentrations of 0.1, 1, 10, 100 and 1000 μM CuO and TiO 2 on common duckweed (Lemna minor L.). Both nano and bulk CuO particles caused changes in the structure and function of treated plants. The number of fronds and colonies decreased by as much as 78%, the length of roots and fronds decreased by 99% and 14%, respectively. Furthermore, photochemical efficiency was reduced by up to 35%, and the activities of antioxidative enzymes guaiacol peroxidase, ascorbate peroxidase and glutathione reductase increased by more than 240%. The altered physiological state of the CuO exposed plants was also reflected in the elevated occurrence of necrosis and bleaching in the duckweed colonies. Nano sized particles of CuO proved more phytotoxic than bulk particles, and the effects of both studied CuO sizes were concentration dependent. On the other hand, both bulk and nano sized particles of TiO 2 caused no severe phytotoxic effects, there was no concentration dependence and they could be considered as non-harmful to common duckweed. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  13. Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin-glycan interactions in clathrin-independent endocytosis.

    Science.gov (United States)

    Mathew, Mohit P; Donaldson, Julie G

    2018-05-11

    Clathrin-independent endocytosis (CIE) is a form of endocytosis that lacks a defined cytoplasmic machinery. Here, we asked whether glycan interactions, acting from the outside, could be a part of that endocytic machinery. We show that the perturbation of global cellular patterns of protein glycosylation by modulation of metabolic flux affects CIE. Interestingly, these changes in glycosylation had cargo-specific effects. For example, in HeLa cells, GlcNAc treatment, which increases glycan branching, increased major histocompatibility complex class I (MHCI) internalization but inhibited CIE of the glycoprotein CD59 molecule (CD59). The effects of knocking down the expression of galectin 3, a carbohydrate-binding protein and an important player in galectin-glycan interactions, were also cargo-specific and stimulated CD59 uptake. By contrast, inhibition of all galectin-glycan interactions by lactose inhibited CIE of both MHCI and CD59. None of these treatments affected clathrin-mediated endocytosis, implying that glycosylation changes specifically affect CIE. We also found that the galectin lattice tailors membrane fluidity and cell spreading. Furthermore, changes in membrane dynamics mediated by the galectin lattice affected macropinocytosis, an altered form of CIE, in HT1080 cells. Our results suggest that glycans play an important and nuanced role in CIE, with each cargo being affected uniquely by alterations in galectin and glycan profiles and their interactions. We conclude that galectin-driven effects exist on a continuum from stimulatory to inhibitory, with distinct CIE cargo proteins having unique response landscapes and with different cell types starting at different positions on these conceptual landscapes.

  14. Quantitative analysis of receptor-mediated uptake and pro-apoptotic activity of mistletoe lectin-1 by high content imaging.

    Science.gov (United States)

    Beztsinna, N; de Matos, M B C; Walther, J; Heyder, C; Hildebrandt, E; Leneweit, G; Mastrobattista, E; Kok, R J

    2018-02-09

    Ribosome inactivating proteins (RIPs) are highly potent cytotoxins that have potential as anticancer therapeutics. Mistletoe lectin 1 (ML1) is a heterodimeric cytotoxic protein isolated from European Mistletoe and belongs to RIP class II. The aim of this project was to systematically study ML1 cell binding, endocytosis pathway(s), subcellular processing and apoptosis activation. For this purpose, state of the art cell imaging equipment and automated image analysis algorithms were used. ML1 displayed very fast binding to sugar residues on the membrane and energy-dependent uptake in CT26 cells. The co-staining with specific antibodies and uptake blocking experiments revealed involvement of both clathrin-dependent and -independent pathways in ML1 endocytosis. Co-localization studies demonstrated the toxin transport from early endocytic vesicles to Golgi network; a retrograde road to the endoplasmic reticulum. The pro-apoptotic and antiproliferative activity of ML1 were shown in time lapse movies and subsequently quantified. ML1 cytotoxicity was less affected in multidrug resistant tumor cell line 4T1 in contrast to commonly used chemotherapeutic drug (ML1 resistance index 6.9 vs 13.4 for doxorubicin; IC 50 : ML1 1.4 ng/ml vs doxorubicin 24000 ng/ml). This opens new opportunities for the use of ML1 as an alternative treatment in multidrug resistant cancers.

  15. Different bulk and active bacterial communities in cryoconite from the margin and interior of the Greenland ice sheet

    DEFF Research Database (Denmark)

    Stibal, Marek; Schostag, Morten; Cameron, Karen A.

    2015-01-01

    composition of cryoconite over a melt season at two contrasting sites at the margin and in the interior of the Greenland ice sheet, using sequence analysis and quantitative polymerase chain reaction of coextracted 16S rDNA and rRNA. Significant differences were found between bulk (rDNA) and potentially active...

  16. Hook-up of GluA2, GRIP and liprin-α for cholinergic muscarinic receptor-dependent LTD in the hippocampus

    Directory of Open Access Journals (Sweden)

    Wu Long-Jun

    2009-06-01

    Full Text Available Abstract The molecular mechanism underlying muscarinic acetylcholine receptor-dependent LTD (mAChR-LTD in the hippocampus is less studied. In a recent study, a novel mechanism is described. The induction of mAChR-LTD required the activation of protein tyrosine phosphatase (PTP, and the expression was mediated by AMPA receptor endocytosis via interactions between GluA2, GRIP and liprin-α. The hook-up of these proteins may result in the recruitment of leukocyte common antigen-related receptor (LAR, a PTP that is known to be involved in AMPA receptor trafficking. Interestingly, the similar molecular interaction cannot be applied to mGluR-LTD, despite the fact that the same G-protein involved in LTD is activated by both mAChR and mGluR. This discovery provides key molecular insights for cholinergic dependent cognitive function, and mAChR-LTD can serve as a useful cellular model for studying the roles of cholinergic mechanism in learning and memory.

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

    Directory of Open Access Journals (Sweden)

    de la Vega Michelle

    2011-12-01

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

  18. Electro-optical modeling of bulk heterojunction solar cells

    Science.gov (United States)

    Kirchartz, Thomas; Pieters, Bart E.; Taretto, Kurt; Rau, Uwe

    2008-11-01

    We introduce a model for charge separation in bulk heterojunction solar cells that combines exciton transport to the interface between donor and acceptor phases with the dissociation of the bound electron/hole pair. We implement this model into a standard semiconductor device simulator, thereby creating a convenient method to simulate the optical and electrical characteristics of a bulk heterojunction solar cell with a commercially available program. By taking into account different collection probabilities for the excitons in the polymer and the fullerene, we are able to reproduce absorptance, internal and external quantum efficiency, as well as current/voltage curves of bulk heterojunction solar cells. We further investigate the influence of mobilities of the free excitons as well as the mobilities of the free charge carriers on the performance of bulk heterojunction solar cells. We find that, in general, the highest efficiencies are achieved with the highest mobilities. However, an optimum finite mobility of free charge carriers can result from a large recombination velocity at the contacts. In contrast, Langevin-type of recombination cannot lead to finite optimum mobilities even though this mechanism has a strong dependence on the free carrier mobilities.

  19. Superconducting bulk magnet for maglev vehicle: Stable levitation performance above permanent magnet guideway

    International Nuclear Information System (INIS)

    Deng, Z.; Zheng, J.; Li, J.; Ma, G.; Lu, Y.; Zhang, Y.; Wang, S.; Wang, J.

    2008-01-01

    High-temperature superconducting (HTS) maglev vehicle is well known as one of the most potential applications of bulk high-temperature superconductors (HTSCs) in transported levitation system. Many efforts have promoted the practice of the HTS maglev vehicle in people's life by enhancing the load capability and stability. Besides improving the material performance of bulk HTSC and optimizing permanent magnet guideway (PMG), magnetization method of bulk HTSC is also very effective for more stable levitation. Up to now, applied onboard bulk HTSCs are directly magnetized by field cooling above the PMG for the present HTS maglev test vehicles or prototypes in China, Germany, Russia, Brazil, and Japan. By the direct-field-cooling-magnetization (DFCM) over PMG, maglev performances of the bulk HTSCs are mainly depended on the PMG's magnetic field. However, introducing HTS bulk magnet into the HTS maglev system breaks this dependence, which is magnetized by other non-PMG magnetic field. The feasibility of this HTS bulk magnet for maglev vehicle is investigated in the paper. The HTS bulk magnet is field-cooling magnetized by a Field Control Electromagnets Workbench (FCEW), which produces a constant magnetic field up to 1 T. The levitation and guidance forces of the HTS bulk magnet over PMG with different trapped flux at 15 mm working height (WH) were measured and compared with that by DFCM in the same applied PMG magnetic field at optimal field-cooling height (FCH) 30 mm, WH 15 mm. It is found that HTS bulk magnet can also realize a stable levitation above PMG. The trapped flux of HTS bulk magnet is easily controllable by the charging current of FCEW, which implies the maglev performances of HTS bulk magnet above PMG will be adjustable according to the practical requirement. The more trapped flux HTS bulk magnet will lead to bigger guidance force and smaller repulsion levitation force above PMG. In the case of saturated trapped flux for experimental HTS bulk magnet, it is

  20. Superconducting bulk magnet for maglev vehicle: Stable levitation performance above permanent magnet guideway

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Z.; Zheng, J.; Li, J.; Ma, G.; Lu, Y.; Zhang, Y.; Wang, S. [Applied Superconductivity Laboratory, Southwest Jiaotong University, Chengdu 610031 (China); Wang, J. [Applied Superconductivity Laboratory, Southwest Jiaotong University, Chengdu 610031 (China)], E-mail: jsywang@home.swjtu.edu.cn

    2008-06-15

    High-temperature superconducting (HTS) maglev vehicle is well known as one of the most potential applications of bulk high-temperature superconductors (HTSCs) in transported levitation system. Many efforts have promoted the practice of the HTS maglev vehicle in people's life by enhancing the load capability and stability. Besides improving the material performance of bulk HTSC and optimizing permanent magnet guideway (PMG), magnetization method of bulk HTSC is also very effective for more stable levitation. Up to now, applied onboard bulk HTSCs are directly magnetized by field cooling above the PMG for the present HTS maglev test vehicles or prototypes in China, Germany, Russia, Brazil, and Japan. By the direct-field-cooling-magnetization (DFCM) over PMG, maglev performances of the bulk HTSCs are mainly depended on the PMG's magnetic field. However, introducing HTS bulk magnet into the HTS maglev system breaks this dependence, which is magnetized by other non-PMG magnetic field. The feasibility of this HTS bulk magnet for maglev vehicle is investigated in the paper. The HTS bulk magnet is field-cooling magnetized by a Field Control Electromagnets Workbench (FCEW), which produces a constant magnetic field up to 1 T. The levitation and guidance forces of the HTS bulk magnet over PMG with different trapped flux at 15 mm working height (WH) were measured and compared with that by DFCM in the same applied PMG magnetic field at optimal field-cooling height (FCH) 30 mm, WH 15 mm. It is found that HTS bulk magnet can also realize a stable levitation above PMG. The trapped flux of HTS bulk magnet is easily controllable by the charging current of FCEW, which implies the maglev performances of HTS bulk magnet above PMG will be adjustable according to the practical requirement. The more trapped flux HTS bulk magnet will lead to bigger guidance force and smaller repulsion levitation force above PMG. In the case of saturated trapped flux for experimental HTS bulk

  1. Bulk oil clauses

    International Nuclear Information System (INIS)

    Gough, N.

    1993-01-01

    The Institute Bulk Oil Clauses produced by the London market and the American SP-13c Clauses are examined in detail in this article. The duration and perils covered are discussed, and exclusions, adjustment clause 15 of the Institute Bulk Oil Clauses, Institute War Clauses (Cargo), and Institute Strikes Clauses (Bulk Oil) are outlined. (UK)

  2. Carbon nanotubes grown on bulk materials and methods for fabrication

    Science.gov (United States)

    Menchhofer, Paul A [Clinton, TN; Montgomery, Frederick C [Oak Ridge, TN; Baker, Frederick S [Oak Ridge, TN

    2011-11-08

    Disclosed are structures formed as bulk support media having carbon nanotubes formed therewith. The bulk support media may comprise fibers or particles and the fibers or particles may be formed from such materials as quartz, carbon, or activated carbon. Metal catalyst species are formed adjacent the surfaces of the bulk support material, and carbon nanotubes are grown adjacent the surfaces of the metal catalyst species. Methods employ metal salt solutions that may comprise iron salts such as iron chloride, aluminum salts such as aluminum chloride, or nickel salts such as nickel chloride. Carbon nanotubes may be separated from the carbon-based bulk support media and the metal catalyst species by using concentrated acids to oxidize the carbon-based bulk support media and the metal catalyst species.

  3. Stoichiometric balance of protein copy numbers is measurable and functionally significant in a protein-protein interaction network for yeast endocytosis.

    Science.gov (United States)

    Holland, David O; Johnson, Margaret E

    2018-03-01

    Stoichiometric balance, or dosage balance, implies that proteins that are subunits of obligate complexes (e.g. the ribosome) should have copy numbers expressed to match their stoichiometry in that complex. Establishing balance (or imbalance) is an important tool for inferring subunit function and assembly bottlenecks. We show here that these correlations in protein copy numbers can extend beyond complex subunits to larger protein-protein interactions networks (PPIN) involving a range of reversible binding interactions. We develop a simple method for quantifying balance in any interface-resolved PPINs based on network structure and experimentally observed protein copy numbers. By analyzing such a network for the clathrin-mediated endocytosis (CME) system in yeast, we found that the real protein copy numbers were significantly more balanced in relation to their binding partners compared to randomly sampled sets of yeast copy numbers. The observed balance is not perfect, highlighting both under and overexpressed proteins. We evaluate the potential cost and benefits of imbalance using two criteria. First, a potential cost to imbalance is that 'leftover' proteins without remaining functional partners are free to misinteract. We systematically quantify how this misinteraction cost is most dangerous for strong-binding protein interactions and for network topologies observed in biological PPINs. Second, a more direct consequence of imbalance is that the formation of specific functional complexes depends on relative copy numbers. We therefore construct simple kinetic models of two sub-networks in the CME network to assess multi-protein assembly of the ARP2/3 complex and a minimal, nine-protein clathrin-coated vesicle forming module. We find that the observed, imperfectly balanced copy numbers are less effective than balanced copy numbers in producing fast and complete multi-protein assemblies. However, we speculate that strategic imbalance in the vesicle forming module

  4. The polyene antimycotics nystatin and filipin disrupt the plasma membrane, whereas natamycin inhibits endocytosis in germinating conidia of Penicillium discolor

    NARCIS (Netherlands)

    Leeuwen, van M.R.; Golovina, E.A.; Dijksterhuis, J.

    2009-01-01

    To investigate the differences in membrane permeability and the effect on endocytosis of the polyene antimycotics nystatin, filipin and natamycin on germinating fungal conidia. Methods and Results: The model system was Penicillium discolor, a food spoilage fungus. Filipin resulted in

  5. Clathrin-dependent internalization of the angiotensin II AT₁A receptor links receptor internalization to COX-2 protein expression in rat aortic vascular smooth muscle cells.

    Science.gov (United States)

    Morinelli, Thomas A; Walker, Linda P; Velez, Juan Carlos Q; Ullian, Michael E

    2015-02-05

    The major effects of Angiotensin II (AngII) in vascular tissue are mediated by AngII AT1A receptor activation. Certain effects initiated by AT1A receptor activation require receptor internalization. In rat aortic vascular smooth muscle cells (RASMC), AngII stimulates cyclooxygenase 2 protein expression. We have previously shown this is mediated by β-arrestin-dependent receptor internalization and NF-κB activation. In this study, a specific inhibitor of clathrin-mediated endocytosis (CME), pitstop-2, was used to test the hypothesis that clathrin-dependent internalization of activated AT1A receptor mediates NF-κB activation and subsequent cyclooxygenase 2 expression. Radioligand binding assays, real time qt-PCR and immunoblotting were used to document the effects of pitstop-2 on AngII binding and signaling in RASMC. Laser scanning confocal microscopy (LSCM) was used to image pitstop-2׳s effects on AT1 receptor/GFP internalization in HEK-293 cells and p65 NF-κB nuclear localization in RASMC. Pitstop-2 significantly inhibited internalization of AT1A receptor (44.7% ± 3.1% Control vs. 13.2% ± 8.3% Pitstop-2; n=3) as determined by radioligand binding studies in RASMC. Studies utilizing AT1A receptor/GFP expressed in HEK 293 cells and LSCM confirmed these findings. Pitstop-2 significantly inhibited AngII-induced p65 NF-κB phosphorylation and nuclear localization, COX-2 message and protein expression in RASMC without altering activation of p42/44 ERK or TNFα signaling. Pitstop-2, a specific inhibitor of clathrin-mediated endocytosis, confirms that internalization of activated AT1A receptor mediates AngII activation of cyclooxygenase 2 expression in RASMC. These data provide support for additional intracellular signaling pathways activated through β-arrestin mediated internalization of G protein-coupled receptors, such as AT1A receptors. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Monte Carlo calculations and neutron spectrometry in quantitative prompt gamma neutron activation analysis (PGNAA) of bulk samples using an isotopic neutron source

    International Nuclear Information System (INIS)

    Spyrou, N.M.; Awotwi-Pratt, J.B.; Williams, A.M.

    2004-01-01

    An activation analysis facility based on an isotopic neutron source (185 GBq 241 Am/Be) which can perform both prompt and cyclic activation analysis on bulk samples, has been used for more than 20 years in many applications including 'in vivo' activation analysis and the determination of the composition of bio-environmental samples, such as, landfill waste and coal. Although the comparator method is often employed, because of the variety in shape, size and elemental composition of these bulk samples, it is often difficult and time consuming to construct appropriate comparator samples for reference. One of the obvious problems is the distribution and energy of the neutron flux in these bulk and comparator samples. In recent years, it was attempted to adopt the absolute method based on a monostandard and to make calculations using a Monte Carlo code (MCNP4C2) to explore this further. In particular, a model of the irradiation facility has been made using the MCNP4C2 code in order to investigate the factors contributing to the quantitative determination of the elemental concentrations through prompt gamma neutron activation analysis (PGNAA) and most importantly, to estimate how the neutron energy spectrum and neutron dose vary with penetration depth into the sample. This simulation is compared against the scattered and transmitted neutron energy spectra that are experimentally and empirically determined using a portable neutron spectrometry system. (author)

  7. Uniform TiO2 nanoparticles induce apoptosis in epithelial cell lines in a size-dependent manner.

    Science.gov (United States)

    Sun, Qingqing; Ishii, Takayuki; Kanehira, Koki; Sato, Takeshi; Taniguchi, Akiyoshi

    2017-05-02

    The size of titanium dioxide (TiO 2 ) nanoparticles is a vital parameter that determines their cytotoxicity. However, most reported studies have employed irregular shapes and sizes of TiO 2 nanoparticles, as it is difficult to produce nanoparticles of suitable sizes for research. We produced good model TiO 2 nanoparticles of uniform shape and size for use in studying their cytotoxicity. In this work, spherical, uniform polyethylene glycol-modified TiO 2 (TiO 2 -PEG) nanoparticles of differing sizes (100, 200, and 300 nm) were prepared using the sol-gel method. A size-dependent decrease in cell viability was observed with increasing nanoparticle size. Furthermore, apoptosis was found to be positively associated with nanoparticle size, as evidenced by an increase in caspase-3 activity with increasing nanoparticle size. Larger nanoparticles exhibited higher cellular uptake, suggesting that larger nanoparticles more strongly induce apoptosis. In addition, the cellular uptake of different sizes of nanoparticles was energy dependent, suggesting that there are size-dependent uptake pathways. We found that 100 and 200 nm (but not 300 nm) nanoparticles were taken up via clathrin-mediated endocytosis. These results utilizing uniform nanoparticles suggest that the size-dependent cytotoxicity of nanoparticles involves active cellular uptake, caspase-3 activation, and apoptosis in the epithelial cell line (NCI-H292). These findings will hopefully aid in the future design and safe use of nanoparticles.

  8. Nuclear Matter Bulk Parameter Scales and Correlations

    International Nuclear Information System (INIS)

    Santos, B. M.; Delfino, A.; Dutra, M.; Lourenço, O.

    2015-01-01

    We study the arising of correlations among some isovector bulk parameters in nonrelativistic and relativistic hadronic mean-field models. For the former, we investigate correlations in the nonrelativistic (NR) limit of relativistic point-coupling models. We provide analytical correlations, for the NR limit model, between the symmetry energy and its derivatives, namely, the symmetry energy slope, curvature, skewness and fourth order derivative, discussing the conditions in which they are linear ones. We also show that some correlations presented in the NR limit model are reproduced for relativistic models presenting cubic and quartic self-interactions in its scalar field. As a direct application of such linear correlations, we remark its association with possible crossing points in the density dependence of the linearly correlated bulk parameter. (author)

  9. Bulk-Fill Resin Composites

    DEFF Research Database (Denmark)

    Benetti, Ana Raquel; Havndrup-Pedersen, Cæcilie; Honoré, Daniel

    2015-01-01

    the restorative procedure. The aim of this study, therefore, was to compare the depth of cure, polymerization contraction, and gap formation in bulk-fill resin composites with those of a conventional resin composite. To achieve this, the depth of cure was assessed in accordance with the International Organization...... for Standardization 4049 standard, and the polymerization contraction was determined using the bonded-disc method. The gap formation was measured at the dentin margin of Class II cavities. Five bulk-fill resin composites were investigated: two high-viscosity (Tetric EvoCeram Bulk Fill, SonicFill) and three low......-viscosity (x-tra base, Venus Bulk Fill, SDR) materials. Compared with the conventional resin composite, the high-viscosity bulk-fill materials exhibited only a small increase (but significant for Tetric EvoCeram Bulk Fill) in depth of cure and polymerization contraction, whereas the low-viscosity bulk...

  10. Big bang nucleosynthesis constraints on bulk neutrinos

    International Nuclear Information System (INIS)

    Goh, H.S.; Mohapatra, R.N.

    2002-01-01

    We examine the constraints imposed by the requirement of successful nucleosynthesis on models with one large extra hidden space dimension and a single bulk neutrino residing in this dimension. We solve the Boltzmann kinetic equation for the thermal distribution of the Kaluza-Klein modes and evaluate their contribution to the energy density at the big bang nucleosynthesis epoch to constrain the size of the extra dimension R -1 ≡μ and the parameter sin 2 2θ which characterizes the mixing between the active and bulk neutrinos

  11. Temperature measurements in small holes drilled in superconducting bulk during pulsed field magnetization

    Science.gov (United States)

    Fujishiro, H.; Naito, T.; Furuta, D.; Kakehata, K.

    2010-11-01

    The time dependence of the temperatures T(z, t) has been measured along the thickness direction z in several drilled holes in a superconducting bulk during pulsed field magnetization (PFM) and the heat generation and heat transfer in the bulk have been discussed. In the previous paper [H. Fujishiro, S. Kawaguchi, K. Kakehata, A. Fujiwara, T. Tateiwa, T. Oka, Supercond. Sci. Technol. 19 (2006) S540], we calculated the T(z, t) profiles in the bulk by solving a three-dimensional heat-diffusion equation to reproduce the measured T(t) on the bulk surface; the heat generation took place adiabatically and the calculated T(z, t) was isothermal along the z direction. In this study, the measured T(z, t) at the top surface was higher than that at the bottom surface just after the pulse field application at t < 0.5 s, and then became isothermal with increasing time. These results suggest that the magnetic flux intrudes inhomogeneously into the bulk from the edge of the top surface and the periphery at the early stage. The inhomogeneous magnetic flux intrusion and the flux trap during PFM change depending on the strength of the pulsed field and the pulse number in the successive pulse field application.

  12. Antioxidant activity of oregano, parsley, and olive mill wastewaters in bulk oils and oil-in-water emulsions enriched in fish oil.

    Science.gov (United States)

    Jimenez-Alvarez, D; Giuffrida, F; Golay, P A; Cotting, C; Lardeau, A; Keely, Brendan J

    2008-08-27

    The antioxidant activity of oregano, parsley, olive mill wastewaters (OMWW), Trolox, and ethylenediaminetetraacetic acid (EDTA) was evaluated in bulk oils and oil-in-water (o/w) emulsions enriched with 5% tuna oil by monitoring the formation of hydroperoxides, hexanal, and t-t-2,4-heptadienal in samples stored at 37 degrees C for 14 days. In bulk oil, the order of antioxidant activity was, in decreasing order (p oregano > parsley > EDTA > Trolox. The antioxidant activity in o/w emulsion followed the same order except that EDTA was as efficient an antioxidant as OMWW. In addition, the total phenolic content, the radical scavenging properties, the reducing capacity, and the iron chelating activity of OMWW, parsley, and oregano extracts were determined by the Folin-Ciocalteau, oxygen radical absorbance capacity, ferric reducing antioxidant power, and iron(II) chelating activity assays, respectively. The antioxidant activity of OMWW, parsley, and oregano in food systems was related to their total phenolic content and radical scavenging capacity but not to their ability to chelate iron in vitro. OMWW was identified as a promising source of antioxidants to retard lipid oxidation in fish oil-enriched food products.

  13. Single-cell vs. bulk activity properties of coastal bacterioplankton over an annual cycle in a temperate ecosystem.

    Science.gov (United States)

    Morán, Xosé Anxelu G; Calvo-Díaz, Alejandra

    2009-01-01

    The connections between single-cell activity properties of heterotrophic planktonic bacteria and whole community metabolism are still poorly understood. Here, we show flow cytometry single-cell analysis of membrane-intact (live), high nucleic acid (HNA) content and actively respiring (CTC+) bacteria with samples collected monthly during 2006 in northern Spain coastal waters. Bulk activity was assessed by measuring 3H-Leucine incorporation and specific growth rates. Consistently, different single-cell relative abundances were found, with 60-100% for live, 30-84% for HNA and 0.2-12% for CTC+ cells. Leucine incorporation rates (2-153 pmol L(-1) h(-1)), specific growth rates (0.01-0.29 day(-1)) and the total and relative abundances of the three single-cell groups showed marked seasonal patterns. Distinct depth distributions during summer stratification and different relations with temperature, chlorophyll and bacterial biovolume suggest the existence of different controlling factors on each single-cell property. Pooled leucine incorporation rates were similarly correlated with the abundance of all physiological groups, while specific growth rates were only substantially explained by the percentage of CTC+ cells. However, the ability to reduce CTC proved notably better than the other two single-cell properties at predicting bacterial bulk rates within seasons, suggesting a tight linkage between bacterial individual respiration and biomass production at the community level.

  14. Photocarrier dynamics in monolayer phosphorene and bulk black phosphorus.

    Science.gov (United States)

    Zereshki, Peymon; Wei, Yaqing; Ceballos, Frank; Bellus, Matthew Z; Lane, Samuel D; Pan, Shudi; Long, Run; Zhao, Hui

    2018-06-13

    We report a combined theoretical and experimental study on photocarrier dynamics in monolayer phosphorene and bulk black phosphorus. Samples of monolayer phosphorene and bulk black phosphorus were fabricated by mechanical exfoliation, identified according to their reflective contrasts, and protected by covering them with hexagonal boron nitride layers. Photocarrier dynamics in these samples was studied by an ultrafast pump-probe technique. The photocarrier lifetime of monolayer phosphorene was found to be about 700 ps, which is about 9 times longer than that of bulk black phosphorus. This trend was reproduced in our calculations based on ab initio nonadiabatic molecular dynamics combined with time-domain density functional theory in the Kohn-Sham representation, and can be attributed to the smaller bandgap and stronger nonadiabatic coupling in bulk. The transient absorption response was also found to be dependent on the sample orientation with respect to the pump polarization, which is consistent with the previously reported anisotropic absorption of phosphorene. In addition, an oscillating component of the differential reflection signal at early probe delays was observed in the bulk sample and was attributed to the layer-breathing phonon mode with an energy of about 1 meV and a decay time of about 1.35 ps. These results provide valuable information for application of monolayer phosphorene in optoelectronics.

  15. Direct determination of bulk etching rate for LR-115-II solid state ...

    Indian Academy of Sciences (India)

    The thickness of the removed layer of the LR-115-II solid state nuclear track detector during etching is measured directly with a rather precise instrument. Dependence of bulk etching rate on temperature of the etching solution is investigated. It has been found that the bulk etching rate is 3.2 m/h at 60°C in 2.5 N NaOH of ...

  16. Effects of receptor-mediated endocytosis and tubular protein composition on volume retention in experimental glomerulonephritis

    DEFF Research Database (Denmark)

    Kastner, Christian; Pohl, Marcus; Sendeski, Mauricio

    2009-01-01

    Human glomerulonephritis (GN) is characterized by sustained proteinuria, sodium retention, hypertension, and edema formation. Increasing quantities of filtered protein enter the renal tubule, where they may alter epithelial transport functions. Exaggerated endocytosis and consequent protein...... overload may affect proximal tubules, but intrinsic malfunction of distal epithelia has also been reported. A straightforward assignment to a particular tubule segment causing salt retention in GN is still controversial. We hypothesized that 1) trafficking and surface expression of major transporters...

  17. Is the bulk mode conversion important in high density helicon plasma?

    Energy Technology Data Exchange (ETDEWEB)

    Isayama, Shogo; Hada, Tohru [Interdisciplinary Graduate School of Engineering Sciences, Kyushu University, 6-1 Kasuga-Kohen, Kasuga, Fukuoka 816-8580 (Japan); Shinohara, Shunjiro [Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588 (Japan); Tanikawa, Takao [Research Institute of Science and Technology, Tokai University 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292 (Japan)

    2016-06-15

    In a high-density helicon plasma production process, a contribution of Trivelpiece-Gould (TG) wave for surface power deposition is widely accepted. The TG wave can be excited either due to an abrupt density gradient near the plasma edge (surface conversion) or due to linear mode conversion from the helicon wave in a density gradient in the bulk region (bulk mode conversion). By numerically solving the boundary value problem of linear coupling between the helicon and the TG waves in a background with density gradient, we show that the efficiency of the bulk mode conversion strongly depends on the dissipation included in the plasma, and the bulk mode conversion is important when the dissipation is small. Also, by performing FDTD simulation, we show the time evolution of energy flux associated with the helicon and the TG waves.

  18. Temperature dependent charge transport studies across thermodynamic glass transition in P3HT:PCBM bulk heterojunction: insight from J-V and impedance spectroscopy

    Science.gov (United States)

    Sarkar, Atri; Rahaman, Abdulla Bin; Banerjee, Debamalya

    2018-03-01

    Temperature dependent charge transport properties of P3HT:PCBM bulk heterojunction are analysed by dc and ac measurements under dark conditions across a wide temperature range of 110-473 K, which includes the thermodynamic glass transition temperature (Tg ˜320 K) of the system. A change from Ohmic conduction to space charge limited current conduction at higher (⩾1.2 V) applied bias voltages above  ⩾200 K is observed from J-V characteristics. From capacitance-voltage (C-V) measurement at room temperature, the occurrence of a peak near the built-in voltage is observed below the dielectric relaxation frequency, originating from the competition between drift and diffusion driven motions of charges. Carrier concentration (N) is calculated from C-V measurements taken at different temperatures. Room temperature mobility values at various applied bias voltages are in accordance with that obtained from transient charge extraction by linearly increasing voltage measurement. Sample impedance is measured over five decades of frequency across temperature range by using lock-in detection. This data is used to extract temperature dependence of carrier mobility (μ), and dc conductivity (σ_dc ) which is low frequency extrapolation of ac conductivity. An activation energy of  ˜126 meV for the carrier hopping process at the metal-semiconductor interface is estimated from temperature dependence of σ_dc . Above T g, μ levels off to a constant value, whereas σ_dc starts to decrease after a transition knee at T g that can be seen as a combined effect of changes in μ and N. All these observed changes across T g can be correlated to enhanced polymer motion above the glass transition.

  19. Bulk density calculations from prompt gamma ray yield

    International Nuclear Information System (INIS)

    Naqvi, A.A.; Nagadi, M.M.; Al-Amoudi, O.S.B.; Maslehuddin, M.

    2006-01-01

    Full text: The gamma ray yield from a Prompt Gamma ray Neutron Activation Analysis (PGNAA) setup is a linear function of element concentration and neutron flux in a the sample with constant bulk density. If the sample bulk density varies as well, then the element concentration and the neutron flux has a nonlinear correlation with the gamma ray yield [1]. The measurement of gamma ray yield non-linearity from samples and a standard can be used to estimate the bulk density of the samples. In this study the prompt gamma ray yield from Blast Furnace Slag, Fly Ash, Silica Fumes and Superpozz cements samples have been measured as a function of their calcium and silicon concentration using KFUPM accelerator-based PGNAA setup [2]. Due to different bulk densities of the blended cement samples, the measured gamma ray yields have nonlinear correlation with calcium and silicon concentration of the samples. The non-linearity in the yield was observed to increase with gamma rays energy and element concentration. The bulk densities of the cement samples were calculated from ratio of gamma ray yield from blended cement and that from a Portland cement standard. The calculated bulk densities have good agreement with the published data. The result of this study will be presented

  20. Integration of bulk piezoelectric materials into microsystems

    Science.gov (United States)

    Aktakka, Ethem Erkan

    Bulk piezoelectric ceramics, compared to deposited piezoelectric thin-films, provide greater electromechanical coupling and charge capacity, which are highly desirable in many MEMS applications. In this thesis, a technology platform is developed for wafer-level integration of bulk piezoelectric substrates on silicon, with a final film thickness of 5-100microm. The characterized processes include reliable low-temperature (200°C) AuIn diffusion bonding and parylene bonding of bulk-PZT on silicon, wafer-level lapping of bulk-PZT with high-uniformity (+/-0.5microm), and low-damage micro-machining of PZT films via dicing-saw patterning, laser ablation, and wet-etching. Preservation of ferroelectric and piezoelectric properties is confirmed with hysteresis and piezo-response measurements. The introduced technology offers higher material quality and unique advantages in fabrication flexibility over existing piezoelectric film deposition methods. In order to confirm the preserved bulk properties in the final film, diaphragm and cantilever beam actuators operating in the transverse-mode are designed, fabricated and tested. The diaphragm structure and electrode shapes/sizes are optimized for maximum deflection through finite-element simulations. During tests of fabricated devices, greater than 12microm PP displacement is obtained by actuation of a 1mm2 diaphragm at 111kHz with integration of a 50-80% efficient power management IC, which incorporates a supply-independent bias circuitry, an active diode for low-dropout rectification, a bias-flip system for higher efficiency, and a trickle battery charger. The overall system does not require a pre-charged battery, and has power consumption of <1microW in active-mode (measured) and <5pA in sleep-mode (simulated). Under lg vibration at 155Hz, a 70mF ultra-capacitor is charged from OV to 1.85V in 50 minutes.

  1. Overexpression of Myo1e in mouse podocytes enhances cellular endocytosis, migration, and adhesion.

    Science.gov (United States)

    Jin, Xia; Wang, Wenjing; Mao, Jianhua; Shen, Huijun; Fu, Haidong; Wang, Xia; Gu, Weizhong; Liu, Aimin; Yu, Huimin; Shu, Qiang; Du, Lizhong

    2014-02-01

    Podocytes are a terminally differentiated and highly specialized cell type in the glomerulus that forms a crucial component of the glomerular filtration barrier. Recently, Myo1e was identified in the podocytes of glomeruli. Myo1e podocyte-specific knockout mice exhibit proteinuria, podocyte foot process effacement, glomerular basement membrane disorganization, signs of chronic renal injury, and kidney inflammation. After overexpression of Myo1e in a conditionally immortalized mouse podocyte cell line (MPC5), podocyte migration was evaluated via transwell assay, endocytosis was evaluated using FITC-transferrin, and adhesion was evaluated using a detachment assay after puromycin aminonucleoside treatment. Myo1e overexpression significantly increased the adherence of podocytes. ANOVA analysis indicated significant differences for cell adhesion between the overexpression and control groups (overexpression vs. control, t = 11.3199, P = 0.005; overexpression vs. negative control, t = 12.0570, P = 0.0006). Overexpression of Myo1e inhibited puromycin aminonucleoside-induced podocyte detachment, and the number of cells remaining on the bottom of the culture plate increased. Cell migration was enhanced in Myo1e-overexpressing podocytes in the transwell migration assay. Internalization of FITC-transferrin also increased in Myo1e-overexpressing podocytes relative to control cells. Overexpression of Myo1e can enhance podocyte migration ability, endocytosis, and attachment to the glomerular basement membrane. Restoration of Myo1e expression in podocytes may therefore strengthen their functional integrity against environmental and mechanical injury. © 2013 Wiley Periodicals, Inc.

  2. Mechanical properties of nano and bulk Fe pillars using molecular dynamics and dislocation dynamics simulation

    Directory of Open Access Journals (Sweden)

    S. K. Deb Nath

    2017-10-01

    Full Text Available Using molecular dynamics simulation, tension and bending tests of a Fe nanopillar are carried out to obtain its Young’s modulus and yield strength. Then the comparative study of Young’s modulus and yield strength of a Fe nanopillar under bending and tension are carried out varying its diameter in the range of diameter 1-15nm. We find out the reasons why bending Young’s modulus and yield strength of a Fe nanopillar are higher than those of tension Young’s modulus and yield strength of a Fe nanopillar. Using the mobility parameters of bulk Fe from the experimental study [N. Urabe and J. Weertman, Materials Science and Engineering 18, 41 (1975], its temperature dependent stress-strain relationship, yield strength and strain hardening modulus are obtained from the dislocation dynamics simulations. Strain rate dependent yield strength and strain hardening modulus of bulk Fe pillars under tension are studied. Temperature dependent creep behaviors of bulk Fe pillars under tension are also studied. To verify the soundness of the present dislocation dynamics studies of the mechanical properties of bulk Fe pillars under tension, the stress vs. strain relationship and dislocation density vs. strain of bulk Fe pillars obtained by us are compared with the published results obtained by S. Queyreau, G. Monnet, and B. Devincre, International Journal of Plasticity 25, 361 (2009.

  3. Ordered bulk degradation via autophagy

    DEFF Research Database (Denmark)

    Dengjel, Jörn; Kristensen, Anders Riis; Andersen, Jens S

    2008-01-01

    During amino acid starvation, cells undergo macroautophagy which is regarded as an unspecific bulk degradation process. Lately, more and more organelle-specific autophagy subtypes such as reticulophagy, mitophagy and ribophagy have been described and it could be shown, depending on the experimental...... at proteasomal and lysosomal degradation ample cross-talk between the two degradation pathways became evident. Degradation via autophagy appeared to be ordered and regulated at the protein complex/organelle level. This raises several important questions such as: can macroautophagy itself be specific and what...

  4. STATUS and DIRECTION OF THE BULK VITRIFICATION PROGRAM FOR THE SUPPLEMENTAL TREATMENT OF LOW ACTIVITY TANK WASTE AT HANFORD

    International Nuclear Information System (INIS)

    RAYMOND, R.E.

    2005-01-01

    The DOE Office of River Protection (ORP) is managing a program at the Hanford site that will retrieve and treat more than 200 million liters (53 million gal.) of radioactive waste stored in underground storage tanks. The waste was generated over the past 50 years as part of the nation's defense programs. The project baseline calls for the waste to be retrieved from the tanks and partitioned to separate the highly radioactive constituents from the large volumes of chemical waste. These highly radioactive components will be vitrified into glass logs in the Waste Treatment Plant (WTP), temporarily stored on the Hanford Site, and ultimately disposed of as high-level waste in the offsite national repository. The less radioactive chemical waste, referred to as low-activity waste (LAW), is also planned to be vitrified by the WTP, and then disposed of in approved onsite trenches. However, additional treatment capacity is required in order to complete the pretreatment and immobilization of the tank waste by 2028, which represents a Tri-Party Agreement milestone. To help ensure that the treatment milestones will be met, the Supplemental Treatment Program was undertaken. The program, managed by CH2M HILL Hanford Group, Inc., involves several sub-projects each intended to supplement part of the treatment of waste being designed into the WTP. This includes the testing, evaluation, design, and deployment of supplemental LAW treatment and immobilization technologies, retrieval and treatment of mixed TRU waste stored in the Hanford Tanks, and supplemental pre-treatment. Applying one or more supplemental treatment technologies to the LAW has several advantages, including providing additional processing capacity, reducing the planned loading on the WTP, and reducing the need for double-shell tank space for interim storage of LAW. In fiscal year 2003, three potential supplemental treatment technologies were evaluated including grout, steam reforming and bulk vitrification using AMEC

  5. Gene Transfer in Eukaryotic Cells Using Activated Dendrimers

    Science.gov (United States)

    Dennig, Jörg

    Gene transfer into eukaryotic cells plays an important role in cell biology. Over the last 30 years a number of transfection methods have been developed to mediate gene transfer into eukaryotic cells. Classical methods include co-precipitation of DNA with calcium phosphate, charge-dependent precipitation of DNA with DEAE-dextran, electroporation of nucleic acids, and formation of transfection complexes between DNA and cationic liposomes. Gene transfer technologies based on activated PAMAM-dendrimers provide another class of transfection reagents. PAMAM-dendrimers are highly branched, spherical molecules. Activation of newly synthesized dendrimers involves hydrolytic removal of some of the branches, and results in a molecule with a higher degree of flexibility. Activated dendrimers assemble DNA into compact structures via charge interactions. Activated dendrimer - DNA complexes bind to the cell membrane of eukaryotic cells, and are transported into the cell by non-specific endocytosis. A structural model of the activated dendrimer - DNA complex and a potential mechanism for its uptake into cells will be discussed.

  6. Accelerating universes driven by bulk particles

    International Nuclear Information System (INIS)

    Brito, F.A.; Cruz, F.F.; Oliveira, J.F.N.

    2005-01-01

    We consider our universe as a 3d domain wall embedded in a 5d dimensional Minkowski space-time. We address the problem of inflation and late time acceleration driven by bulk particles colliding with the 3d domain wall. The expansion of our universe is mainly related to these bulk particles. Since our universe tends to be permeated by a large number of isolated structures, as temperature diminishes with the expansion, we model our universe with a 3d domain wall with increasing internal structures. These structures could be unstable 2d domain walls evolving to fermi-balls which are candidates to cold dark matter. The momentum transfer of bulk particles colliding with the 3d domain wall is related to the reflection coefficient. We show a nontrivial dependence of the reflection coefficient with the number of internal dark matter structures inside the 3d domain wall. As the population of such structures increases the velocity of the domain wall expansion also increases. The expansion is exponential at early times and polynomial at late times. We connect this picture with string/M-theory by considering BPS 3d domain walls with structures which can appear through the bosonic sector of a five-dimensional supergravity theory

  7. Properties of Bulk Sintered Silver As a Function of Porosity

    Energy Technology Data Exchange (ETDEWEB)

    Wereszczak, Andrew A [ORNL; Vuono, Daniel J [ORNL; Wang, Hsin [ORNL; Ferber, Mattison K [ORNL; Liang, Zhenxian [ORNL

    2012-06-01

    This report summarizes a study where various properties of bulk-sintered silver were investigated over a range of porosity. This work was conducted within the National Transportation Research Center's Power Device Packaging project that is part of the DOE Vehicle Technologies Advanced Power Electronics and Electric Motors Program. Sintered silver, as an interconnect material in power electronics, inherently has porosity in its produced structure because of the way it is made. Therefore, interest existed in this study to examine if that porosity affected electrical properties, thermal properties, and mechanical properties because any dependencies could affect the intended function (e.g., thermal transfer, mechanical stress relief, etc.) or reliability of that interconnect layer and alter how its performance is modeled. Disks of bulk-sintered silver were fabricated using different starting silver pastes and different sintering conditions to promote different amounts of porosity. Test coupons were harvested out of the disks to measure electrical resistivity and electrical conductivity, thermal conductivity, coefficient of thermal expansion, elastic modulus, Poisson's ratio, and yield stress. The authors fully recognize that the microstructure of processed bulk silver coupons may indeed not be identical to the microstructure produced in thin (20-50 microns) layers of sintered silver. However, measuring these same properties with such a thin actual structure is very difficult, requires very specialized specimen preparation and unique testing instrumentation, is expensive, and has experimental shortfalls of its own, so the authors concluded that the herein measured responses using processed bulk sintered silver coupons would be sufficient to determine acceptable values of those properties. Almost all the investigated properties of bulk sintered silver changed with porosity content within a range of 3-38% porosity. Electrical resistivity, electrical conductivity

  8. Formation, structure, and evolution of boiling nucleus and interfacial tension between bulk liquid phase and nucleus

    Science.gov (United States)

    Wang, Xiao-Dong; Peng, Xiao-Feng; Tian, Yong; Wang, Bu-Xuan

    2005-05-01

    In this paper, the concept of the molecular free path is introduced to derive a criterion distinguishing active molecules from inactive molecules in liquid phase. A concept of the critical aggregation concentration (CAC) of active molecules is proposed to describe the physical configuration before the formation of a nucleus during vapor-liquid phase transition. All active molecules exist as monomers when the concentration of active molecules is lower than CAC, while the active molecules will generate aggregation once the concentration of the active molecules reaches CAC. However, these aggregates with aggregation number, N, smaller than five can steadily exist in bulk phase. The other excess active molecules can only produce infinite aggregation and form a critical nucleus of vapor-liquid phase transition. Without any outer perturbation the state point of CAC corresponds to the critical superheated or supercooled state. Meanwhile, a model of two-region structure of a nucleus is proposed to describe nucleus evolution. The interfacial tension between bulk liquid phase and nucleus is dependent of the density gradient in the transition region and varies with the structure change of the transition region. With the interfacial tension calculated using this model, the predicted nucleation rate is very close to the experimental measurement. Furthermore, this model and associated analysis provides solid theoretical evidences to clarify the definition of nucleation rate and understand nucleation phenomenon with the insight into the physical nature.

  9. Size-Dependent Regulation of Intracellular Trafficking of Polystyrene Nanoparticle-Based Drug-Delivery Systems.

    Science.gov (United States)

    Wang, Ting; Wang, Lu; Li, Xiaoming; Hu, Xingjie; Han, Yuping; Luo, Yao; Wang, Zejun; Li, Qian; Aldalbahi, Ali; Wang, Lihua; Song, Shiping; Fan, Chunhai; Zhao, Yun; Wang, Maolin; Chen, Nan

    2017-06-07

    Nanoparticles (NPs) have shown great promise as intracellular imaging probes or nanocarriers and are increasingly being used in biomedical applications. A detailed understanding of how NPs get "in and out" of cells is important for developing new nanomaterials with improved selectivity and less cytotoxicity. Both physical and chemical characteristics have been proven to regulate the cellular uptake of NPs. However, the exocytosis process and its regulation are less explored. Herein, we investigated the size-regulated endocytosis and exocytosis of carboxylated polystyrene (PS) NPs. PS NPs with a smaller size were endocytosed mainly through the clathrin-dependent pathway, whereas PS NPs with a larger size preferred caveolae-mediated endocytosis. Furthermore, our results revealed exocytosis of larger PS NPs and tracked the dynamic process at the single-particle level. These results indicate that particle size is a key factor for the regulation of intracellular trafficking of NPs and provide new insight into the development of more effective cellular nanocarriers.

  10. Nusselt number and bulk temperature in turbulent Rayleigh-Bénard convection

    Science.gov (United States)

    Bodenschatz, Eberhard; Weiss, Stephan; Shishkina, Olga; International CollaborationTurbulence Research Collaboration

    2017-11-01

    We present an algorithm to calculate the Nusselt number (Nu) in measurements of the heat transport in turbulent Rayleigh-Bénard convection under general non-Oberbeck-Boussinesq (NOB) conditions. We further critically analyze the different ways to evaluate the dependences of Nu over the Rayleigh number (Ra) and show the sensitivity of these dependences to the reference temperatures in the bulk, top and bottom boundary layers (BLs). Finally we propose a method to predict the bulk temperature and a way to calculate the reference temperatures of the top and bottom BLs and validate them against the Göttingen measurements. The work is supported by the Max Planck Society and the Deutsche Forschungsgemeinschaft (DFG) under the Grant Sh 405/4 - Heisenberg fellowship.

  11. Perovskite oxides: Oxygen electrocatalysis and bulk structure

    Science.gov (United States)

    Carbonio, R. E.; Fierro, C.; Tryk, D.; Scherson, D.; Yeager, Ernest

    1987-01-01

    Perovskite type oxides were considered for use as oxygen reduction and generation electrocatalysts in alkaline electrolytes. Perovskite stability and electrocatalytic activity are studied along with possible relationships of the latter with the bulk solid state properties. A series of compounds of the type LaFe(x)Ni1(-x)O3 was used as a model system to gain information on the possible relationships between surface catalytic activity and bulk structure. Hydrogen peroxide decomposition rate constants were measured for these compounds. Ex situ Mossbauer effect spectroscopy (MES), and magnetic susceptibility measurements were used to study the solid state properties. X ray photoelectron spectroscopy (XPS) was used to examine the surface. MES has indicated the presence of a paramagnetic to magnetically ordered phase transition for values of x between 0.4 and 0.5. A correlation was found between the values of the MES isomer shift and the catalytic activity for peroxide decomposition. Thus, the catalytic activity can be correlated to the d-electron density for the transition metal cations.

  12. Temperature dependence of water-water and ion-water correlations in bulk water and electrolyte solutions probed by femtosecond elastic second harmonic scattering

    Science.gov (United States)

    Chen, Yixing; Dupertuis, Nathan; Okur, Halil I.; Roke, Sylvie

    2018-06-01

    The temperature dependence of the femtosecond elastic second harmonic scattering (fs-ESHS) response of bulk light and heavy water and their electrolyte solutions is presented. We observe clear temperature dependent changes in the hydrogen (H)-bond network of water that show a decrease in the orientational order of water with increasing temperature. Although D2O has a more structured H-bond network (giving rise to more fs-ESHS intensity), the relative temperature dependence is larger in H2O. The changes are interpreted in terms of the symmetry of H-bonds and are indicators of nuclear quantum effects. Increasing the temperature in electrolyte solutions decreases the influence of the total electrostatic field from ions on the water-water correlations, as expected from Debye-Hückel theory, since the Debye length becomes longer. The effects are, however, 1.9 times (6.3 times) larger than those predicted for H2O (D2O). Since fs-ESHS responses can be computed from known molecular coordinates, our observations provide a unique opportunity to refine quantum mechanical models of water.

  13. Bulk-boundary correlators in the hermitian matrix model and minimal Liouville gravity

    International Nuclear Information System (INIS)

    Bourgine, Jean-Emile; Ishiki, Goro; Rim, Chaiho

    2012-01-01

    We construct the one matrix model (MM) correlators corresponding to the general bulk-boundary correlation numbers of the minimal Liouville gravity (LG) on the disc. To find agreement between both discrete and continuous approach, we investigate the resonance transformation mixing boundary and bulk couplings. It leads to consider two sectors, depending on whether the matter part of the LG correlator is vanishing due to the fusion rules. In the vanishing case, we determine the explicit transformation of the boundary couplings at the first order in bulk couplings. In the non-vanishing case, no bulk-boundary resonance is involved and only the first order of pure boundary resonances have to be considered. Those are encoded in the matrix polynomials determined in our previous paper. We checked the agreement for the bulk-boundary correlators of MM and LG in several non-trivial cases. In this process, we developed an alternative method to derive the boundary resonance encoding polynomials.

  14. Holographic complexity for time-dependent backgrounds

    Energy Technology Data Exchange (ETDEWEB)

    Momeni, Davood, E-mail: davoodmomeni78@gmail.com [Eurasian International Center for Theoretical Physics and Department of General Theoretical Physics, Eurasian National University, Astana 010008 (Kazakhstan); Faizal, Mir, E-mail: mirfaizalmir@googlemail.com [Irving K. Barber School of Arts and Sciences, University of British Columbia, Okanagan, 3333 University Way, Kelowna, British Columbia V1V 1V7 (Canada); Department of Physics and Astronomy, University of Lethbridge, Lethbridge, Alberta, T1K 3M4 (Canada); Bahamonde, Sebastian, E-mail: sebastian.beltran.14@ucl.ac.uk [Department of Mathematics, University College London, Gower Street, London, WC1E 6BT (United Kingdom); Myrzakulov, Ratbay [Eurasian International Center for Theoretical Physics and Department of General Theoretical Physics, Eurasian National University, Astana 010008 (Kazakhstan)

    2016-11-10

    In this paper, we will analyze the holographic complexity for time-dependent asymptotically AdS geometries. We will first use a covariant zero mean curvature slicing of the time-dependent bulk geometries, and then use this co-dimension one spacelike slice of the bulk spacetime to define a co-dimension two minimal surface. The time-dependent holographic complexity will be defined using the volume enclosed by this minimal surface. This time-dependent holographic complexity will reduce to the usual holographic complexity for static geometries. We will analyze the time-dependence as a perturbation of the asymptotically AdS geometries. Thus, we will obtain time-dependent asymptotically AdS geometries, and we will calculate the holographic complexity for such time-dependent geometries.

  15. Time-dependent, glucose-regulated Arabidopsis Regulator of G-protein Signaling 1 network

    Directory of Open Access Journals (Sweden)

    Dinesh Kumar Jaiswal

    2016-04-01

    Full Text Available Plants lack 7-transmembrane, G-protein coupled receptors (GPCRs because the G alpha subunit of the heterotrimeric G protein complex is “self-activating”—meaning that it spontaneously exchanges bound GDP for GTP without the need of a GPCR. In lieu of GPCRs, most plants have a seven transmembrane receptor-like regulator of G-protein signaling (RGS protein, a component of the complex that keeps G-protein signaling in its non-activated state. The addition of glucose physically uncouples AtRGS1 from the complex through specific endocytosis leaving the activated G protein at the plasma membrane. The complement of proteins in the AtRGS1/G-protein complex over time from glucose-induced endocytosis was profiled by immunoprecipitation coupled to mass spectrometry (IP-MS. A total of 119 proteins in the AtRGS1 complex were identified. Several known interactors of the complex were identified, thus validating the approach, but the vast majority (93/119 were not known previously. AtRGS1 protein interactions were dynamically modulated by d-glucose. At low glucose levels, the AtRGS1 complex is comprised of proteins involved in transport, stress and metabolism. After glucose application, the AtRGS1 complex rapidly sheds many of these proteins and recruits other proteins involved in vesicular trafficking and signal transduction. The profile of the AtRGS1 components answers several questions about the type of coat protein and vesicular trafficking GTPases used in AtRGS1 endocytosis and the function of endocytic AtRGS1.

  16. Palladium diffusion into bulk copper via the (100) surface

    Energy Technology Data Exchange (ETDEWEB)

    Bussmann, E; Kellogg, G L [Sandia National Laboratories, Albuquerque, NM 87185 (United States); Sun, J; Pohl, K [Department of Physics and Materials Science Program, University of New Hampshire, Durham, NH 03824 (United States)

    2009-08-05

    Using low-energy electron microscopy, we measure the diffusion of Pd into bulk Cu at the Cu(100) surface. Interdiffusion is tracked by measuring the dissolution of the Cu(100)-c(2 x 2)-Pd surface alloy during annealing (T>240 deg. C). The activation barrier for Pd diffusion from the surface alloy into the bulk is determined to be (1.8 +- 0.6) eV. During annealing, we observe the growth of a new layer of Cu near step edges. Under this new Cu layer, dilute Pd remaining near the surface develops a layered structure similar to the Cu{sub 3}Pd L 1{sub 2} bulk alloy phase.

  17. The multimodal magnetoelectric effect in the ring-shaped magnetostrictive-piezoelectric bulk composites

    Science.gov (United States)

    Radchenko, G. S.; Filippov, D. A.; Laletin, V. M.

    2015-11-01

    The theoretical and experimental investigation of the direct magnetoelectric effect in the ring-type structures made of the bulk magnetostrictive-piezoelectric composites has been presented. The analytical expression for the magnetoelectric voltage coefficient has been obtained using the effective parameters method. The frequency dependence of this parameter is also analyzed. The dependence of the resonant frequency and the amplitude of this effect of the geometrical parameters of the ring for the first and second oscillation modes are presented. The experimental investigation of the direct magnetoelectric effect for the ring-type composite specimens consisting of the nickel ferrite spinel-PZT bulk composite is done. The obtained experimental data are in good agreement with the theoretical predictions.

  18. THE OPTIMIZATION OF PLUSH YARNS BULKING PROCESS

    Directory of Open Access Journals (Sweden)

    VINEREANU Adam

    2014-05-01

    Full Text Available This paper presents the experiments that were conducted on the installation of continuous bulking and thermofixing “SUPERBA” type TVP-2S for optimization of the plush yarns bulking process. There were considered plush yarns Nm 6.5/2, made of the fibrous blend of 50% indigenous wool sort 41 and 50% PES. In the first stage, it performs a thermal treatment with a turboprevaporizer at a temperature lower than thermofixing temperature, at atmospheric pressure, such that the plush yarns - deposed in a freely state on a belt conveyor - are uniformly bulking and contracting. It was followed the mathematical modeling procedure, working with a factorial program, rotatable central composite type, and two independent variables. After analyzing the parameters that have a direct influence on the bulking degree, there were selected the pre-vaporization temperature (coded x1,oC and the velocity of belt inside pre-vaporizer (coded x 2, m/min. As for the dependent variable, it was chosen the plush yarn diameter (coded y, mm. There were found the coordinates of the optimal point, and then this pair of values was verified in practice. These coordinates are: x1optim= 90oC and x 2optim= 6.5 m/min. The conclusion is that the goal was accomplished: it was obtained a good cover degree f or double-plush carpets by reducing the number of tufts per unit surface.

  19. Dynamic analysis of bulk-fill composites: Effect of food-simulating liquids.

    Science.gov (United States)

    Eweis, Ahmed Hesham; Yap, Adrian U-Jin; Yahya, Noor Azlin

    2017-10-01

    This study investigated the effect of food simulating liquids on visco-elastic properties of bulk-fill restoratives using dynamic mechanical analysis. One conventional composite (Filtek Z350 [FZ]), two bulk-fill composites (Filtek Bulk-fill [FB] and Tetric N Ceram [TN]) and a bulk-fill giomer (Beautifil-Bulk Restorative [BB]) were evaluated. Specimens (12 × 2 × 2mm) were fabricated using customized stainless steel molds. The specimens were light-cured, removed from their molds, finished, measured and randomly divided into six groups. The groups (n = 10) were conditioned in the following mediums for 7 days at 37°C: air (control), artificial saliva (SAGF), distilled water, 0.02N citric acid, heptane, 50% ethanol-water solution. Specimens were assessed using dynamic mechanical testing in flexural three-point bending mode and their respective mediums at 37°C and a frequency range of 0.1-10Hz. The distance between the supports were fixed at 10mm and an axial load of 5N was employed. Data for elastic modulus, viscous modulus and loss tangent were subjected to ANOVA/Tukey's tests at significance level p food-simulating liquids on the visco-elastic properties of bulk-fill composites was material and medium dependent. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Relation between Raman backscattering from droplets and bulk water: Effect of refractive index dispersion

    Science.gov (United States)

    Plakhotnik, Taras; Reichardt, Jens

    2018-03-01

    A theoretical framework is presented that permits investigations of the relation between inelastic backscattering from microparticles and bulk samples of Raman-active materials. It is based on the Lorentz reciprocity theorem and no fundamental restrictions concerning the microparticle shape apply. The approach provides a simple and intuitive explanation for the enhancement of the differential backscattering cross-section in particles in comparison to bulk. The enhancement factor for scattering of water droplets in the diameter range from 0 to 60 μm (vitally important for the a priori measurement of liquid water content of warm clouds with spectroscopic Raman lidars) is about a factor of 1.2-1.6 larger (depending on the size of the sphere) than an earlier study has shown. The numerical calculations are extended to 1000 μm and demonstrate that dispersion of the refractive index of water becomes an important factor for spheres larger than 100 μm. The physics of the oscillatory phenomena predicted by the simulations is explained.

  1. HIV internalization into oral and genital epithelial cells by endocytosis and macropinocytosis leads to viral sequestration in the vesicles

    Science.gov (United States)

    Yasen, Aizezi; Herrera, Rossana; Rosbe, Kristina; Lien, Kathy; Tugizov, Sharof M.

    2018-01-01

    Recently, we showed that HIV-1 is sequestered, i.e., trapped, in the intracellular vesicles of oral and genital epithelial cells. Here, we investigated the mechanisms of HIV-1 sequestration in vesicles of polarized tonsil, foreskin and cervical epithelial cells. HIV-1 internalization into epithelial cells is initiated by multiple entry pathways, including clathrin-, caveolin/lipid raft-associated endocytosis and macropinocytosis. Inhibition of HIV-1 attachment to galactosylceramide and heparan sulfate proteoglycans, and virus endocytosis and macropinocytosis reduced HIV-1 sequestration by 30–40%. T-cell immunoglobulin and mucin domain 1 (TIM-1) were expressed on the apical surface of polarized tonsil, cervical and foreskin epithelial cells. However, TIM-1-associated HIV-1 macropinocytosis and sequestration were detected mostly in tonsil epithelial cells. Sequestered HIV-1 was resistant to trypsin, pronase, and soluble CD4, indicating that the sequestered virus was intracellular. Inhibition of HIV-1 intraepithelial sequestration and elimination of vesicles containing virus in the mucosal epithelium may help in the prevention of HIV-1 mucosal transmission. PMID:29277006

  2. Cetuximab in combination with anti-human IgG antibodies efficiently down-regulates the EGF receptor by macropinocytosis

    International Nuclear Information System (INIS)

    Berger, Christian; Madshus, Inger Helene; Stang, Espen

    2012-01-01

    The monoclonal antibody C225 (Cetuximab) blocks binding of ligand to the epidermal growth factor receptor (EGFR). In addition, it is known that incubation with C225 induces endocytosis of the EGFR. This endocytosis has previously been shown to be increased when C225 is combined with an additional monoclonal anti-EGFR antibody. However, the effects of antibody combinations on EGFR activation, endocytosis, trafficking and degradation have been unclear. By binding a secondary antibody to the C225-EGFR complex, we here demonstrate that a combination of antibodies can efficiently internalize and degrade the EGFR. Although the combination of antibodies activated the EGFR kinase and induced ubiquitination of the EGFR, the kinase activity was not required for internalization of the EGFR. In contrast to EGF-induced EGFR down-regulation, the antibody combination efficiently degraded the EGFR without initiating downstream proliferative signaling. The antibody-induced internalization of EGFR was found not to depend on clathrin and/or dynamin, but depended on actin polymerization, suggesting induction of macropinocytosis. Macropinocytosis may cause internalization of large membrane areas, and this could explain the highly efficient internalization of the EGFR induced by combination of antibodies. -- Highlight: ► Cetuximab induced endocytosis of EGFR increases upon combination with anti-human IgG. ► Antibody combination causes internalization of EGFR by macropinocytosis. ► Antibody-induced internalization of EGFR is independent of EGFR kinase activity. ► Antibody combination may have a zipper effect and cross-link EGFRs on neighboring cells.

  3. Distinguishing bulk traps and interface states in deep-level transient spectroscopy

    International Nuclear Information System (INIS)

    Coelho, A V P; Adam, M C; Boudinov, H

    2011-01-01

    A new method for the distinction of discrete bulk deep levels and interface states related peaks in deep-level transient spectroscopy spectra is proposed. The measurement of two spectra using different reverse voltages while keeping pulse voltage fixed causes different peak maximum shifts in each case: for a reverse voltage modulus increase, a bulk deep-level related peak maximum will remain unchanged or shift towards lower temperatures while only interface states related peak maximum will be able to shift towards higher temperatures. This method has the advantage of being non-destructive and also works in the case of bulk traps with strong emission rate dependence on the electric field. Silicon MOS capacitors and proton implanted GaAs Schottky diodes were employed to experimentally test the method.

  4. Transport logistics in pollen tubes.

    Science.gov (United States)

    Chebli, Youssef; Kroeger, Jens; Geitmann, Anja

    2013-07-01

    Cellular organelles move within the cellular volume and the effect of the resulting drag forces on the liquid causes bulk movement in the cytosol. The movement of both organelles and cytosol leads to an overall motion pattern called cytoplasmic streaming or cyclosis. This streaming enables the active and passive transport of molecules and organelles between cellular compartments. Furthermore, the fusion and budding of vesicles with and from the plasma membrane (exo/endocytosis) allow for transport of material between the inside and the outside of the cell. In the pollen tube, cytoplasmic streaming and exo/endocytosis are very active and fulfill several different functions. In this review, we focus on the logistics of intracellular motion and transport processes as well as their biophysical underpinnings. We discuss various modeling attempts that have been performed to understand both long-distance shuttling and short-distance targeting of organelles. We show how the combination of mechanical and mathematical modeling with cell biological approaches has contributed to our understanding of intracellular transport logistics.

  5. Ratio of bulk to shear viscosity in a quasigluon plasma: from weak to strong coupling

    CERN Document Server

    Bluhm, M; Redlich, K

    2012-01-01

    The ratio of bulk to shear viscosity is expected to exhibit a different behaviour in weakly and in strongly coupled systems. This can be expressed by the dependence of the ratio on the squared sound velocity. In the high temperature QCD plasma at small running coupling, the viscosity ratio is uniquely determined by a quadratic dependence on the conformality measure, whereas in certain strongly coupled and nearly conformal theories this dependence is linear. Employing an effective kinetic theory of quasiparticle excitations with medium-modified dispersion relation, we analyze the ratio of bulk to shear viscosity of the gluon plasma. We show that in this approach the viscosity ratio comprises both dependencies found by means of weak coupling perturbative and strong coupling holographic techniques.

  6. The effect of surface-bulk potential difference on the kinetics of intercalation in core-shell active cathode particles

    Science.gov (United States)

    Kazemiabnavi, Saeed; Malik, Rahul; Orvananos, Bernardo; Abdellahi, Aziz; Ceder, Gerbrand; Thornton, Katsuyo

    2018-04-01

    Surface modification of active cathode particles is commonly observed in battery research as either a surface phase evolving during the cycling process, or intentionally engineered to improve capacity retention, rate capability, and/or thermal stability of the cathode material. Here, a continuum-scale model is developed to simulate the galvanostatic charge/discharge of a cathode particle with core-shell heterostructure. The particle is assumed to be comprised of a core material encapsulated by a thin layer of a second phase that has a different open-circuit voltage. The effect of the potential difference between the surface and bulk phases (Ω) on the kinetics of lithium intercalation and the galvanostatic charge/discharge profiles is studied at different values of Ω, C-rates, and exchange current densities. The difference between the Li chemical potential in the surface and bulk phases of the cathode particle results in a concentration difference between these two phases. This leads to a charge/discharge asymmetry in the galvanostatic voltage profiles, causing a decrease in the accessible capacity of the particle. These effects are more significant at higher magnitudes of surface-bulk potential difference. The proposed model provides detailed insight into the kinetics and voltage behavior of the intercalation/de-intercalation processes in core-shell heterostructure cathode particles.

  7. Temperature dependence of the bulk and surface properties of liquid Zn-Cd alloys

    Energy Technology Data Exchange (ETDEWEB)

    Awe, O.E. [University of Ibadan, Department of Physics, Ibadan (Nigeria); Azeez, A.A. [African University of Science and Technology, Abuja (Nigeria)

    2017-05-15

    The effects of temperature on the bulk and surface properties of liquid Zn-Cd alloys have been theoretically investigated, using a combination of self association model, Darken's thermodynamic equation for diffusion, empirical model for viscosity and a statistical mechanics model. The results from this study show that change in temperature resulted in cross-over effects in bulk and surface properties. We also found that with an increase in temperature, a pronounced asymmetry of viscosity isotherm is significantly reduced, and viscosity isotherm exhibited anomalous behaviour. Our results reveal that the homocoordination tendency in Zn-Cd liquid alloys is not strong and reduces with increasing temperature. The study further suggests a pronounced segregation of Cd-atoms at the surface of Zn-Cd liquid alloys and the extent of segregation reduces with temperature. We as well found that, in addition to the reported understanding that size-factor determines the compositional location of asymmetry of the viscosity isotherm, temperature is an operating parameter that has effect, not only on the composition of asymmetry, but also on the magnitude of asymmetry. In all the properties investigated, the most pronounced effect of temperature (52.9 %) is on the viscosity while the least effect (7.1 %) is on the surface tension. (orig.)

  8. Charge based DC compact modeling of bulk FinFET transistor

    Science.gov (United States)

    Cerdeira, A.; Garduño, I.; Tinoco, J.; Ritzenthaler, R.; Franco, J.; Togo, M.; Chiarella, T.; Claeys, C.

    2013-09-01

    Multiple-gate MOSFETs became an industrial reality in the last years. Due to a pragmatic trade-off between CMOS process baselines compatibility, improved performance compared to planar bulk architecture, and cost, bulk FinFETs emerged as the technological solution to provide downscaling for the 14/22 nm technological nodes. In this work, a charge based DC compact model based on the SDDG Model is demonstrated for this new generation of FinFET transistors and describes continuously the transistor characteristics in all operating regions. Validating the model against two bulk FinFET baselines (NMOS, PMOS, various gate lengths and EOT), an excellent agreement is found for transfer and output characteristics (linear and saturation regimes), transconductance/output conductance, and gm/IDS characteristics. Temperature dependence is also taken into account and validated (T range from 25 °C up to 175 °C).

  9. Calculation of thermal neutron self-shielding correction factors for aqueous bulk sample prompt gamma neutron activation analysis using the MCNP code

    International Nuclear Information System (INIS)

    Nasrabadi, M.N.; Jalali, M.; Mohammadi, A.

    2007-01-01

    In this work thermal neutron self-shielding in aqueous bulk samples containing neutron absorbing materials is studied using bulk sample prompt gamma neutron activation analysis (BSPGNAA) with the MCNP code. The code was used to perform three dimensional simulations of a neutron source, neutron detector and sample of various material compositions. The MCNP model was validated against experimental measurements of the neutron flux performed using a BF 3 detector. Simulations were performed to predict thermal neutron self-shielding in aqueous bulk samples containing neutron absorbing solutes. In practice, the MCNP calculations are combined with experimental measurements of the relative thermal neutron flux over the sample's surface, with respect to a reference water sample, to derive the thermal neutron self-shielding within the sample. The proposed methodology can be used for the determination of the elemental concentration of unknown aqueous samples by BSPGNAA where knowledge of the average thermal neutron flux within the sample volume is required

  10. Shiga Toxin Increases Formation of Clathrin-Coated Pits through Syk Kinase

    DEFF Research Database (Denmark)

    Utskarpen, Audrun; Massol, Ramiro; van Deurs, Bo

    2010-01-01

    Clathrin-dependent endocytosis is a main entry mechanism for the glycolipid-binding Shiga toxin (Stx), although clathrin-independent pathways are also involved. Binding of Stx to its receptor Gb3 not only is essential for Stx retrograde transport to the endoplasmic reticulum and toxicity but also...... activates signaling through the tyrosine kinase Syk. We previously described that Syk activity is important for Stx entry, but it remained unclear how this kinase modulates endocytosis of Stx. Here we characterized the effects of Stx and Syk on clathrin-coated pit formation. We found that acute treatment...

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

    DEFF Research Database (Denmark)

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

    1995-01-01

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

  12. Temperature-dependent dielectric and energy-storage properties of Pb(Zr,Sn,Ti)O{sub 3} antiferroelectric bulk ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xuefeng; Liu, Zhen; Xu, Chenhong; Cao, Fei; Wang, Genshui; Dong, Xianlin, E-mail: xldong@mail.sic.ac.cn [Key laboratory of Inorganic Functional Materials and Devices, Shanghai Institute of Ceramics, Chinese Academy of Sciences, 200050, Shanghai (China)

    2016-05-15

    The dielectric and energy-storage properties of Pb{sub 0.99}Nb{sub 0.02}[(Zr{sub 0.60}Sn{sub 0.40}){sub 0.95}Ti{sub 0.05}]{sub 0.98}O{sub 3} (PNZST) bulk ceramics near the antiferroelectric (AFE)-ferroelectric (FE) phase boundary are investigated as a function of temperature. Three characteristic temperatures T{sub 0}, T{sub C}, T{sub 2} are obtained from the dielectric temperature spectrum. At different temperature regions (below T{sub 0}, between T{sub 0} and T{sub C}, and above T{sub C}), three types of hysteresis loops are observed as square double loop, slim loop and linear loop, respectively. The switching fields and recoverable energy density all first increase and then decrease with increasing temperature, and reach their peak values at ∼T{sub 0}. These results provide a convenient method to optimize the working temperature of antiferroelectric electronic devices through testing the temperature dependent dielectric properties of antiferroelectric ceramics.

  13. Profiling Y561-dependent and -independent substrates of CSF-1R in epithelial cells.

    Directory of Open Access Journals (Sweden)

    Melodie L Knowlton

    2010-10-01

    Full Text Available Receptor tyrosine kinases (RTKs activate multiple downstream cytosolic tyrosine kinases following ligand stimulation. SRC family kinases (SFKs, which are recruited to activated RTKs through SH2 domain interactions with RTK autophosphorylation sites, are targets of many subfamilies of RTKs. To date, there has not been a systematic analysis of the downstream substrates of such receptor-activated SFKs. Here, we conducted quantitative mass spectrometry utilizing stable isotope labeling (SILAC analysis to profile candidate SRC-substrates induced by the CSF-1R tyrosine kinase by comparing the phosphotyrosine-containing peptides from cells expressing either CSF-1R or a mutant form of this RTK that is unable to bind to SFKs. This analysis identified previously uncharacterized changes in tyrosine phosphorylation induced by CSF-1R in mammary epithelial cells as well as a set of candidate substrates dependent on SRC recruitment to CSF-1R. Many of these candidates may be direct SRC targets as the amino acids flanking the phosphorylation sites in these proteins are similar to known SRC kinase phosphorylation motifs. The putative SRC-dependent proteins include known SRC substrates as well as previously unrecognized SRC targets. The collection of substrates includes proteins involved in multiple cellular processes including cell-cell adhesion, endocytosis, and signal transduction. Analyses of phosphoproteomic data from breast and lung cancer patient samples identified a subset of the SRC-dependent phosphorylation sites as being strongly correlated with SRC activation, which represent candidate markers of SRC activation downstream of receptor tyrosine kinases in human tumors. In summary, our data reveal quantitative site-specific changes in tyrosine phosphorylation induced by CSF-1R activation in epithelial cells and identify many candidate SRC-dependent substrates phosphorylated downstream of an RTK.

  14. Nanoindentation creep versus bulk compressive creep of dental resin-composites.

    Science.gov (United States)

    El-Safty, S; Silikas, N; Akhtar, R; Watts, D C

    2012-11-01

    To evaluate nanoindentation as an experimental tool for characterizing the viscoelastic time-dependent creep of resin-composites and to compare the resulting parameters with those obtained by bulk compressive creep. Ten dental resin-composites: five conventional, three bulk-fill and two flowable were investigated using both nanoindentation creep and bulk compressive creep methods. For nano creep, disc specimens (15mm×2mm) were prepared from each material by first injecting the resin-composite paste into metallic molds. Specimens were irradiated from top and bottom surfaces in multiple overlapping points to ensure optimal polymerization using a visible light curing unit with output irradiance of 650mW/cm(2). Specimens then were mounted in 3cm diameter phenolic ring forms and embedded in a self-curing polystyrene resin. Following grinding and polishing, specimens were stored in distilled water at 37°C for 24h. Using an Agilent Technologies XP nanoindenter equipped with a Berkovich diamond tip (100nm radius), the nano creep was measured at a maximum load of 10mN and the creep recovery was determined when each specimen was unloaded to 1mN. For bulk compressive creep, stainless steel split molds (4mm×6mm) were used to prepare cylindrical specimens which were thoroughly irradiated at 650mW/cm(2) from multiple directions and stored in distilled water at 37°C for 24h. Specimens were loaded (20MPa) for 2h and unloaded for 2h. One-way ANOVA, Levene's test for homogeneity of variance and the Bonferroni post hoc test (all at p≤0.05), plus regression plots, were used for statistical analysis. Dependent on the type of resin-composite material and the loading/unloading parameters, nanoindentation creep ranged from 29.58nm to 90.99nm and permanent set ranged from 8.96nm to 30.65nm. Bulk compressive creep ranged from 0.47% to 1.24% and permanent set ranged from 0.09% to 0.38%. There was a significant (p=0.001) strong positive non-linear correlation (r(2)=0.97) between bulk

  15. RNASEK Is a V-ATPase-Associated Factor Required for Endocytosis and the Replication of Rhinovirus, Influenza A Virus, and Dengue Virus

    Directory of Open Access Journals (Sweden)

    Jill M. Perreira

    2015-08-01

    Full Text Available Human rhinovirus (HRV causes upper respiratory infections and asthma exacerbations. We screened multiple orthologous RNAi reagents and identified host proteins that modulate HRV replication. Here, we show that RNASEK, a transmembrane protein, was needed for the replication of HRV, influenza A virus, and dengue virus. RNASEK localizes to the cell surface and endosomal pathway and closely associates with the vacuolar ATPase (V-ATPase proton pump. RNASEK is required for endocytosis, and its depletion produces enlarged clathrin-coated pits (CCPs at the cell surface. These enlarged CCPs contain endocytic cargo and are bound by the scissioning GTPase, DNM2. Loss of RNASEK alters the localization of multiple V-ATPase subunits and lowers the levels of the ATP6AP1 subunit. Together, our results show that RNASEK closely associates with the V-ATPase and is required for its function; its loss prevents the early events of endocytosis and the replication of multiple pathogenic viruses.

  16. Constraint-based modeling and kinetic analysis of the Smad dependent TGF-beta signaling pathway.

    Directory of Open Access Journals (Sweden)

    Zhike Zi

    Full Text Available BACKGROUND: Investigation of dynamics and regulation of the TGF-beta signaling pathway is central to the understanding of complex cellular processes such as growth, apoptosis, and differentiation. In this study, we aim at using systems biology approach to provide dynamic analysis on this pathway. METHODOLOGY/PRINCIPAL FINDINGS: We proposed a constraint-based modeling method to build a comprehensive mathematical model for the Smad dependent TGF-beta signaling pathway by fitting the experimental data and incorporating the qualitative constraints from the experimental analysis. The performance of the model generated by constraint-based modeling method is significantly improved compared to the model obtained by only fitting the quantitative data. The model agrees well with the experimental analysis of TGF-beta pathway, such as the time course of nuclear phosphorylated Smad, the subcellular location of Smad and signal response of Smad phosphorylation to different doses of TGF-beta. CONCLUSIONS/SIGNIFICANCE: The simulation results indicate that the signal response to TGF-beta is regulated by the balance between clathrin dependent endocytosis and non-clathrin mediated endocytosis. This model is useful to be built upon as new precise experimental data are emerging. The constraint-based modeling method can also be applied to quantitative modeling of other signaling pathways.

  17. Nicotine Dependence, Physical Activity, and Sedentary Behavior among Adult Smokers.

    Science.gov (United States)

    Loprinzi, Paul D; Walker, Jerome F

    2015-03-01

    Research has previously demonstrated an inverse association between smoking status and physical activity; however, few studies have examined the association between nicotine dependence and physical activity or sedentary behavior. This study examined the association between nicotine dependence and accelerometer-determined physical activity and sedentary behavior. Data from the 2003-2006 National Health and Nutrition Examination Survey (NHANES) were used. A total of 851 adult (≥20 years) smokers wore an accelerometer for ≥4 days and completed the Fagerstrom Test for Nicotine Dependence scale. Regression models were used to examine the association between nicotine dependence and physical activity/sedentary behavior. After adjusting for age, gender, race-ethnicity, poverty level, hypertension, emphysema, bronchitis, body mass index (BMI), cotinine, and accelerometer wear time, smokers 50 + years of age with greater nicotine dependence engaged in more sedentary behavior (β = 11.4, P = 0.02) and less light-intensity physical activity (β = -9.6, P = 0.03) and moderate-to-vigorous physical activity (MVPA; β = -0.14, P = 0.003) than their less nicotine dependent counterparts. Older adults who are more nicotine dependent engage in less physical activity (both MVPA and light-intensity) and more sedentary behavior than their less nicotine dependent counterparts.

  18. Zinc-ion-dependent acid phosphatase exhibits magnesium-ion-dependent myo-inositol-1-phosphatase activity.

    Science.gov (United States)

    Fujimoto, S; Okano, I; Tanaka, Y; Sumida, Y; Tsuda, J; Kawakami, N; Shimohama, S

    1996-06-01

    We have purified bovine brain Zn(2+)-dependent acid phosphatase (Zn(2+)-APase), which requires Zn2+ ions to hydrolyze the substrate p-nitrophenyl phosphate (pNPP) in an acidic environment. The substrate specificity and metal requirement of Zn(2+)-APase at a physiological pH was also studied. The enzyme exhibited hydrolytic activity on myo-inositol-1- and -2-monophosphates, 2'-adenosine monophosphate, 2'-guanosine monophosphate, and the alpha- and beta-glycerophosphates, glucose-1-phosphate, and fructose-6-phosphate in 50 mM Tris-HCl buffer (pH 7.4) in the presence of Mg2+ ions, but not on pNPP and phosphotyrosine. Zn2+, Mn2+ and Co2+ ions were less effective for activation. Among the above substrates, myo-inositol-1-phosphate was the most susceptible to hydrolysis by the enzyme in the presence of 3 mM Mg2+ ions. The enzyme exhibited an optimum pH at around 8 for myo-inositol-1-phosphate in the presence of 3 mM Mg2+ ions. The Mg(2+)-dependent myo-inositol-1-phosphatase activity of the enzyme was significantly inhibited by Li+ ions. The Zn(2+)-dependent p-nitrophenyl phosphatase activity and Mg(2+)-dependent myo-inositol-1-phosphatase activity of the purified enzyme fraction exhibited similar behavior on Sephadex G-100 and Mono Q colomns. These findings suggest that Zn(2+)-APase also exhibits Mg(2+)-dependent myo-inositol-1-phosphatase activity under physiological conditions.

  19. Bulk contribution to magnetotransport properties of low-defect-density Bi2Te3 topological insulator thin films

    Science.gov (United States)

    Ngabonziza, P.; Wang, Y.; Brinkman, A.

    2018-04-01

    An important challenge in the field of topological materials is to carefully disentangle the electronic transport contribution of the topological surface states from that of the bulk. For Bi2Te3 topological insulator samples, bulk single crystals and thin films exposed to air during fabrication processes are known to be bulk conducting, with the chemical potential in the bulk conduction band. For Bi2Te3 thin films grown by molecular beam epitaxy, we combine structural characterization (transmission electron microscopy), chemical surface analysis as function of time (x-ray photoelectron spectroscopy) and magnetotransport analysis to understand the low defect density and record high bulk electron mobility once charge is doped into the bulk by surface degradation. Carrier densities and electronic mobilities extracted from the Hall effect and the quantum oscillations are consistent and reveal a large bulk carrier mobility. Because of the cylindrical shape of the bulk Fermi surface, the angle dependence of the bulk magnetoresistance oscillations is two dimensional in nature.

  20. Distinguishing Between Activated and Non-Activated Eosinophils Using a Microelectrode: Theoretical Investigations of Bulk and Surface Polaritons in Magnetic Multilayers

    Science.gov (United States)

    Lacy, Fred

    In Part I of this dissertation, a whole cell biosensor which can detect the activation state of eosinophils (one of several types of white blood cells) will be developed and tested. This biosensor, which consists of a small gold electrode (50 μm x 50 μm) and a large gold electrode (1.5 cm x 0.5 cm) on a glass substrate, has been fabricated by photolithographic techniques. The eosinophils are known to exhibit different physical properties when they change from the activated state to the non-activated state. Based on some of these property changes, there should be a corresponding change in the measured electrical impedance. In this research, this biosensor will measure the electrical impedance of the eosinophils. This will show that the biosensor can detect the different states of the eosinophils (through the electrical impedance technique). And from these measurements, the different parameters associated with the electrical impedance can be determined. In Part II of this dissertation, a theoretical calculation will be performed in which bulk and surface magnetic polaritons in magnetic materials will be found. A polariton is the coupling of electromagnetic radiation and the elementary excitation of the given material (in our case, a magnetic material). The structure that we will be considering is a periodic semi-infinite material consisting of alternating antiferromagnetic and nonmagnetic layers. An antiferromagnetic material is a material in which individual atoms exhibit magnetic moments, but the overall magnetization of the material is zero because the moments of every other atom are antiparallel. We will use a method known as the transfer matrix technique to find an expression for the dispersion relation of the bulk and surface waves in these materials. Then we will create plots of omega(k) as we vary the geometric configurations of the layers which make-up the magnetic multilayer. We also will calculate the effect of an external magnetic field on these

  1. Activity-dependent modulation of neural circuit synaptic connectivity

    Directory of Open Access Journals (Sweden)

    Charles R Tessier

    2009-07-01

    Full Text Available In many nervous systems, the establishment of neural circuits is known to proceed via a two-stage process; 1 early, activity-independent wiring to produce a rough map characterized by excessive synaptic connections, and 2 subsequent, use-dependent pruning to eliminate inappropriate connections and reinforce maintained synapses. In invertebrates, however, evidence of the activity-dependent phase of synaptic refinement has been elusive, and the dogma has long been that invertebrate circuits are “hard-wired” in a purely activity-independent manner. This conclusion has been challenged recently through the use of new transgenic tools employed in the powerful Drosophila system, which have allowed unprecedented temporal control and single neuron imaging resolution. These recent studies reveal that activity-dependent mechanisms are indeed required to refine circuit maps in Drosophila during precise, restricted windows of late-phase development. Such mechanisms of circuit refinement may be key to understanding a number of human neurological diseases, including developmental disorders such as Fragile X syndrome (FXS and autism, which are hypothesized to result from defects in synaptic connectivity and activity-dependent circuit function. This review focuses on our current understanding of activity-dependent synaptic connectivity in Drosophila, primarily through analyzing the role of the fragile X mental retardation protein (FMRP in the Drosophila FXS disease model. The particular emphasis of this review is on the expanding array of new genetically-encoded tools that are allowing cellular events and molecular players to be dissected with ever greater precision and detail.

  2. Effects of surface and bulk transverse fields on critical behaviour of ferromagnetic films

    International Nuclear Information System (INIS)

    Saber, A.; Lo Russo, S.; Mattei, G.

    2002-02-01

    The influence of surface and bulk transverse fields on the critical behaviour of a ferromagnetic Ising film is studied using the effective field theory based on a single-site cluster method. Surface exchange enhancement is considered and a critical value is obtained. The dependence of the critical uniform transverse field on film thickness, phase diagrams in the fields, critical surface transverse field versus the bulk one, and exchange coupling ratio are presented. (author)

  3. Assessing the dependence of bulk ice properties from probes with anti-shatter tips on environmental conditions

    Science.gov (United States)

    Jackson, Robert C.

    Ice clouds have significant impacts on the Earth's radiative budget. Their radiative impact highly depends on ice cloud microphysical properties. Climate and weather prediction models have to make certain assumptions about how the various processes are represented. Observations of how cloud properties vary with environmental conditions can help evaluate some parameterizations used in models. However, sufficient data are not available to characterize how ice crystal properties vary as a function of environmental conditions. Furthermore, many of these assumptions are derived from historical datasets collected by in situ probes, namely optical array probes that can be contaminated by shattered artifacts generated by large particles shattering on the probe tips and inlets. Therefore this study has two main objectives. Prior estimates of ice crystal size distributions derived from 2D Cloud Probes (2DCs) have been artificially amplified by small ice crystals generated from the shattering of large ice crystals on the probe tips. Although anti-shatter tips and algorithms exist, there is considerable uncertainty in their effectiveness. Therefore, this thesis first examines the differences in ice crystal size distributions, and bulk and optical properties from adjacent 2DCs with standard and anti-shatter tips, and processed with and without anti-shattering algorithms. The measurements were obtained from the National Research Council of Canada Convair-580 during the 2008 Indirect and Semi-Direct Aerosol Campaign (ISDAC) and the National Center for Atmospheric Research C-130 during the 2011 Instrumentation Development and Education in Airborne Science 2011 (IDEAS-2011). The 2DC size distributions are compared with those from the Holographic Detector for Clouds (HOLODEC), which has anti-shatter tips and allows for identification of shattering through spatial statistics. The ratio of the number concentration N of particles with maximum dimensions 125 to 500 mum from the 2DC with

  4. Endocytosis of wheat germ agglutinin binding sites from the cell surface into a tubular endosomal network.

    Science.gov (United States)

    Raub, T J; Koroly, M J; Roberts, R M

    1990-04-01

    By using fluorescence and electron microscopy, the endocytic pathway encountered by cell surface components after they had bound wheat germ agglutinin (WGA) was visualized. The majority of these components are thought to consist of sialylated glycoproteins (HMWAG) that represent a subpopulation of the total cell surface proteins but most of the externally disposed plasma membrane proteins of the cell. Examination of semi-thin sections by medium- and high-voltage electron microscopy revealed the three-dimensional organization of vesicular and tubular endosomes. Binding of either fluorescein isothiocyanate-, horseradish peroxidase-, or ferritin-conjugated WGA to cells at 4 degrees C showed that the HMWAG were distributed uniformly over the cell surface. Warming of surface-labeled cells to 37 degrees C resulted in the endocytosis of WGA into peripheral endosomes via invagination of regions of both coated and uncoated membrane. The peripheral endosome appeared as isolated complexes comprising a vesicular element (300-400 nm diam.) surrounded by and continuous with tubular cisternae (45-60 nm diam.), which did not interconnect the endosomes. After 30 min or more label also became localized in a network of anastomosing tubules (45-60 nm diam.) that were located in the centrosomal region of the cell. Endocytosed WGA-HMWAG complexes did not become associated with cisternae of the Golgi apparatus, although tubular and vesicular endosomes were noted in the vicinity of the trans-Golgi region. The accumulation of WGA-HMWAG in the endosomes within the centrosomal region was inhibited when cells were incubated at 18 degrees C. None of these compartments contained acid phosphatase activity, a result that is consistent with other data that the HMWAG do not pass through lysosomes initially. The kinetics of labeling were consistent with the interpretation that recycling of most of the WGA binding surface glycoproteins occurred rapidly from early peripheral endosomes followed by the

  5. Bulk viscosity of strongly interacting matter in the relaxation time approximation

    Science.gov (United States)

    Czajka, Alina; Hauksson, Sigtryggur; Shen, Chun; Jeon, Sangyong; Gale, Charles

    2018-04-01

    We show how thermal mean field effects can be incorporated consistently in the hydrodynamical modeling of heavy-ion collisions. The nonequilibrium correction to the distribution function resulting from a temperature-dependent mass is obtained in a procedure which automatically satisfies the Landau matching condition and is thermodynamically consistent. The physics of the bulk viscosity is studied here for Boltzmann and Bose-Einstein gases within the Chapman-Enskog and 14-moment approaches in the relaxation time approximation. Constant and temperature-dependent masses are considered in turn. It is shown that, in the small mass limit, both methods lead to the same value of the ratio of the bulk viscosity to its relaxation time. The inclusion of a temperature-dependent mass leads to the emergence of the βλ function in that ratio, and it is of the expected parametric form for the Boltzmann gas, while for the Bose-Einstein case it is affected by the infrared cutoff. This suggests that the relaxation time approximation may be too crude to obtain a reliable form of ζ /τR for gases obeying Bose-Einstein statistics.

  6. The effect of solvent on the morphology of an inkjet printed active layer of bulk heterojunction solar cells

    International Nuclear Information System (INIS)

    Fauzia, Vivi; Umar, Akrajas Ali; Salleh, Muhamad Mat; Yahaya, Muhammad

    2011-01-01

    Bulk heterojunction organic solar cells were fabricated by sandwiching the active layer between indium tin oxide (ITO) and Al electrodes. The active layer used was a blend of poly(3-octylthiophene-2,5-diyl) (P3OT) as the electron donor and (6,6)-phenyl C 71 butyric acid methyl ester (PC 71 BM) as the electron acceptor. The active layer thin films were deposited by an inkjet printing technique. Prior to deposition of the thin films, the active materials were blended in three different solvents. The printed films were annealed at three different temperatures. It was found that the selection of the appropriate solvent and annealing treatment significantly influences the printing process, the morphology of the printed film and subsequently the performance of the solar cell devices

  7. Pulsed field magnetization strategies and the field poles composition in a bulk-type superconducting motor

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Zhen, E-mail: zhen.huang@sjtu.edu.cn [Academy of Information Technology and Electrical Engineering, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Ruiz, H.S., E-mail: dr.harold.ruiz@le.ac.uk [Department of Engineering, University of Leicester, University Road, Leicester LE1 7RH (United Kingdom); Coombs, T.A., E-mail: tac1000@cam.ac.uk [Department of Engineering, University of Cambridge, 9 JJ Thomson Avenue, Cambridge CB3 0FA (United Kingdom)

    2017-03-15

    Highlights: • Different compositions of the magnetic poles have been obtained depending on the relative orientation of the magnetizing coil and the surfaces of the columns of bulks that conform a magnetic pole. • Two bidimensional models accounting for the electromagnetic response of the top and lateral cross sections of three columns of HTS bulks subjected to multiple pulsed magnetic fields have been created. • An extended PFM strategy has been proposed by considering the magnetization of at least three successive columns of HTS bulks per pole. In the extended PFM strategy the area of each one of the poles can be seen increased by a factor of 200%-400% - Abstract: High temperature superconducting (HTS) bulks offer the potential of trapping and maintaining much higher magnetic loading level compared with the conventional permanent magnets used in rotary machines, although the effective magnetization of multiple HTS bulks with different relative orientations over the surface of cylindrical rotors creates new challenges. In this paper, we present the design and numerical validation of the Pulse Field Magnetization (PFM) strategy considered for the magnetization of the four-pole synchronous fully superconducting motor developed at the University of Cambridge. In a first instance, singular columns of up to five HTS bulks aligned over the height of the rotor were subjected to up to three magnetic pulses of 1.5 T peak, and the experimental results have been simulated by considering the electrical and thermal properties of the system in a 2D approach. The entire active surface of the rotor is covered by HTS bulks of approximately the same dimensions, resulting in an uneven distribution of pole areas with at least one of the poles formed by up to 3 columns of magnetized bulks, with relatively the same peaks of trapped magnetic field. Thus, in order to effectively use the entire area of the superconducting rotor, multiple pulsed fields per column have been applied

  8. Pulsed field magnetization strategies and the field poles composition in a bulk-type superconducting motor

    International Nuclear Information System (INIS)

    Huang, Zhen; Ruiz, H.S.; Coombs, T.A.

    2017-01-01

    Highlights: • Different compositions of the magnetic poles have been obtained depending on the relative orientation of the magnetizing coil and the surfaces of the columns of bulks that conform a magnetic pole. • Two bidimensional models accounting for the electromagnetic response of the top and lateral cross sections of three columns of HTS bulks subjected to multiple pulsed magnetic fields have been created. • An extended PFM strategy has been proposed by considering the magnetization of at least three successive columns of HTS bulks per pole. In the extended PFM strategy the area of each one of the poles can be seen increased by a factor of 200%-400% - Abstract: High temperature superconducting (HTS) bulks offer the potential of trapping and maintaining much higher magnetic loading level compared with the conventional permanent magnets used in rotary machines, although the effective magnetization of multiple HTS bulks with different relative orientations over the surface of cylindrical rotors creates new challenges. In this paper, we present the design and numerical validation of the Pulse Field Magnetization (PFM) strategy considered for the magnetization of the four-pole synchronous fully superconducting motor developed at the University of Cambridge. In a first instance, singular columns of up to five HTS bulks aligned over the height of the rotor were subjected to up to three magnetic pulses of 1.5 T peak, and the experimental results have been simulated by considering the electrical and thermal properties of the system in a 2D approach. The entire active surface of the rotor is covered by HTS bulks of approximately the same dimensions, resulting in an uneven distribution of pole areas with at least one of the poles formed by up to 3 columns of magnetized bulks, with relatively the same peaks of trapped magnetic field. Thus, in order to effectively use the entire area of the superconducting rotor, multiple pulsed fields per column have been applied

  9. Neurotrophin release by neurotrophins: Implications for activity-dependent neuronal plasticity

    OpenAIRE

    Canossa, Marco; Griesbeck, Oliver; Berninger, Benedikt; Campana, Gabriele; Kolbeck, Roland; Thoenen, Hans

    1997-01-01

    Neurotrophins, secreted in an activity-dependent manner, are thought to be involved in the activity-dependent refinement of synaptic connections. Here we demonstrate that in hippocampal neurons and the rat pheochromocytoma cell line PC12 application of exogenous neurotrophins induces secretion of neurotrophins, an effect that is mediated by the activation of tyrosine kinase neurotrophin receptors (Trks). Like activity-dependent secretion of neurotrophins, neurotrophin-induced neurotrophin sec...

  10. Puerarin activates endothelial nitric oxide synthase through estrogen receptor-dependent PI3-kinase and calcium-dependent AMP-activated protein kinase

    International Nuclear Information System (INIS)

    Hwang, Yong Pil; Kim, Hyung Gyun; Hien, Tran Thi; Jeong, Myung Ho; Jeong, Tae Cheon; Jeong, Hye Gwang

    2011-01-01

    The cardioprotective properties of puerarin, a natural product, have been attributed to the endothelial nitric oxide synthase (eNOS)-mediated production of nitric oxide (NO) in EA.hy926 endothelial cells. However, the mechanism by which puerarin activates eNOS remains unclear. In this study, we sought to identify the intracellular pathways underlying eNOS activation by puerarin. Puerarin induced the activating phosphorylation of eNOS on Ser1177 and the production of NO in EA.hy926 cells. Puerarin-induced eNOS phosphorylation required estrogen receptor (ER)-mediated phosphatidylinositol 3-kinase (PI3K)/Akt signaling and was reversed by AMP-activated protein kinase (AMPK) and calcium/calmodulin-dependent kinase II (CaMKII) inhibition. Importantly, puerarin inhibited the adhesion of tumor necrosis factor (TNF)-α-stimulated monocytes to endothelial cells and suppressed the TNF-α induced expression of intercellular cell adhesion molecule-1. Puerarin also inhibited the TNF-α-induced nuclear factor-κB activation, which was attenuated by pretreatment with N G -nitro-L-arginine methyl ester, a NOS inhibitor. These results indicate that puerarin stimulates eNOS phosphorylation and NO production via activation of an estrogen receptor-mediated PI3K/Akt- and CaMKII/AMPK-dependent pathway. Puerarin may be useful for the treatment or prevention of endothelial dysfunction associated with diabetes and cardiovascular disease. -- Highlights: ► Puerarin induced the phosphorylation of eNOS and the production of NO. ► Puerarin activated eNOS through ER-dependent PI3-kinase and Ca 2+ -dependent AMPK. ► Puerarin-induced NO was involved in the inhibition of NF-kB activation. ► Puerarin may help for prevention of vascular dysfunction and diabetes.

  11. Cetuximab in combination with anti-human IgG antibodies efficiently down-regulates the EGF receptor by macropinocytosis

    Energy Technology Data Exchange (ETDEWEB)

    Berger, Christian [Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway); Madshus, Inger Helene [Institute of Pathology, University of Oslo, Rikshospitalet, 0027 Oslo (Norway); Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway); Stang, Espen, E-mail: espsta@rr-research.no [Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway)

    2012-12-10

    The monoclonal antibody C225 (Cetuximab) blocks binding of ligand to the epidermal growth factor receptor (EGFR). In addition, it is known that incubation with C225 induces endocytosis of the EGFR. This endocytosis has previously been shown to be increased when C225 is combined with an additional monoclonal anti-EGFR antibody. However, the effects of antibody combinations on EGFR activation, endocytosis, trafficking and degradation have been unclear. By binding a secondary antibody to the C225-EGFR complex, we here demonstrate that a combination of antibodies can efficiently internalize and degrade the EGFR. Although the combination of antibodies activated the EGFR kinase and induced ubiquitination of the EGFR, the kinase activity was not required for internalization of the EGFR. In contrast to EGF-induced EGFR down-regulation, the antibody combination efficiently degraded the EGFR without initiating downstream proliferative signaling. The antibody-induced internalization of EGFR was found not to depend on clathrin and/or dynamin, but depended on actin polymerization, suggesting induction of macropinocytosis. Macropinocytosis may cause internalization of large membrane areas, and this could explain the highly efficient internalization of the EGFR induced by combination of antibodies. -- Highlight: Black-Right-Pointing-Pointer Cetuximab induced endocytosis of EGFR increases upon combination with anti-human IgG. Black-Right-Pointing-Pointer Antibody combination causes internalization of EGFR by macropinocytosis. Black-Right-Pointing-Pointer Antibody-induced internalization of EGFR is independent of EGFR kinase activity. Black-Right-Pointing-Pointer Antibody combination may have a zipper effect and cross-link EGFRs on neighboring cells.

  12. Gas selectivity of SILAR grown CdS nano-bulk junction

    Science.gov (United States)

    Jayakrishnan, R.; Nair, Varun G.; Anand, Akhil M.; Venugopal, Meera

    2018-03-01

    Nano-particles of cadmium sulphide were deposited on cleaned copper substrate by an automated sequential ionic layer adsorption reaction (SILAR) system. The grown nano-bulk junction exhibits Schottky diode behavior. The response of the nano-bulk junction was investigated under oxygen and hydrogen atmospheric conditions. The gas response ratio was found to be 198% for Oxygen and 34% for Hydrogen at room temperature. An increase in the operating temperature of the nano-bulk junction resulted in a decrease in their gas response ratio. A logarithmic dependence on the oxygen partial pressure to the junction response was observed, indicating a Temkin isothermal behavior. Work function measurements using a Kelvin probe demonstrate that the exposure to an oxygen atmosphere fails to effectively separate the charges due to the built-in electric field at the interface. Based on the benefits like simple structure, ease of fabrication and response ratio the studied device is a promising candidate for gas detection applications.

  13. Superparamagnetic poly(methyl methacrylate) nanoparticles surface modified with folic acid presenting cell uptake mediated by endocytosis

    Energy Technology Data Exchange (ETDEWEB)

    Feuser, Paulo Emilio [Federal University of Santa Catarina, Department of Chemical Engineering and Food Engineering (Brazil); Jacques, Amanda Virtuoso [Federal University of Santa Catarina, Department of Clinical Analyses (Brazil); Arévalo, Juan Marcelo Carpio; Rocha, Maria Eliane Merlin [Federal University of Paraná, Department of Biochemistry and Molecular Biology (Brazil); Santos-Silva, Maria Claudia dos [Federal University of Santa Catarina, Department of Clinical Analyses (Brazil); Sayer, Claudia; Araújo, Pedro H. Hermes de, E-mail: pedro.h.araujo@ufsc.br [Federal University of Santa Catarina, Department of Chemical Engineering and Food Engineering (Brazil)

    2016-04-15

    The encapsulation of superparamagnetic nanoparticles (MNPs) in polymeric nanoparticles (NPs) with modified surfaces can improve targeted delivery and induce cell death by hyperthermia. The goals of this study were to synthesize and characterize surface modified superparamagnetic poly(methyl methacrylate) with folic acid (FA) prepared by miniemulsion polymerization (MNPsPMMA-FA) and to evaluate their in vitro cytotoxicity and cellular uptake in non-tumor cells, murine fibroblast (L929) cells and tumor cells that overexpressed folate receptor (FR) β, and chronic myeloid leukemia cells in blast crisis (K562). Lastly, hemolysis assays were performed on human red blood cells. MNPsPMMA-FA presented an average mean diameter of 135 nm and a saturation magnetization (Ms) value of 37 emu/g of iron oxide, as well as superparamagnetic behavior. The MNPsPMMA-FA did not present cytotoxicity in L929 and K562 cells. Cellular uptake assays showed a higher uptake of MNPsPMMA-FA than MNPsPMMA in K562 cells when incubated at 37 °C. On the other hand, MNPsPMMA-FA showed a low uptake when endocytosis mechanisms were blocked at low temperature (4 °C), suggesting that the MNPsPMMA-FA uptake was mediated by endocytosis. High concentrations of MNPsPMMA-FA showed hemocompatibility when incubated for 24 h in human red blood cells. Therefore, our results suggest that these carrier systems can be an excellent alternative in targeted drug delivery via FR.

  14. Superparamagnetic poly(methyl methacrylate) nanoparticles surface modified with folic acid presenting cell uptake mediated by endocytosis

    International Nuclear Information System (INIS)

    Feuser, Paulo Emilio; Jacques, Amanda Virtuoso; Arévalo, Juan Marcelo Carpio; Rocha, Maria Eliane Merlin; Santos-Silva, Maria Claudia dos; Sayer, Claudia; Araújo, Pedro H. Hermes de

    2016-01-01

    The encapsulation of superparamagnetic nanoparticles (MNPs) in polymeric nanoparticles (NPs) with modified surfaces can improve targeted delivery and induce cell death by hyperthermia. The goals of this study were to synthesize and characterize surface modified superparamagnetic poly(methyl methacrylate) with folic acid (FA) prepared by miniemulsion polymerization (MNPsPMMA-FA) and to evaluate their in vitro cytotoxicity and cellular uptake in non-tumor cells, murine fibroblast (L929) cells and tumor cells that overexpressed folate receptor (FR) β, and chronic myeloid leukemia cells in blast crisis (K562). Lastly, hemolysis assays were performed on human red blood cells. MNPsPMMA-FA presented an average mean diameter of 135 nm and a saturation magnetization (Ms) value of 37 emu/g of iron oxide, as well as superparamagnetic behavior. The MNPsPMMA-FA did not present cytotoxicity in L929 and K562 cells. Cellular uptake assays showed a higher uptake of MNPsPMMA-FA than MNPsPMMA in K562 cells when incubated at 37 °C. On the other hand, MNPsPMMA-FA showed a low uptake when endocytosis mechanisms were blocked at low temperature (4 °C), suggesting that the MNPsPMMA-FA uptake was mediated by endocytosis. High concentrations of MNPsPMMA-FA showed hemocompatibility when incubated for 24 h in human red blood cells. Therefore, our results suggest that these carrier systems can be an excellent alternative in targeted drug delivery via FR.

  15. Superparamagnetic poly(methyl methacrylate) nanoparticles surface modified with folic acid presenting cell uptake mediated by endocytosis

    Science.gov (United States)

    Feuser, Paulo Emilio; Jacques, Amanda Virtuoso; Arévalo, Juan Marcelo Carpio; Rocha, Maria Eliane Merlin; dos Santos-Silva, Maria Claudia; Sayer, Claudia; de Araújo, Pedro H. Hermes

    2016-04-01

    The encapsulation of superparamagnetic nanoparticles (MNPs) in polymeric nanoparticles (NPs) with modified surfaces can improve targeted delivery and induce cell death by hyperthermia. The goals of this study were to synthesize and characterize surface modified superparamagnetic poly(methyl methacrylate) with folic acid (FA) prepared by miniemulsion polymerization (MNPsPMMA-FA) and to evaluate their in vitro cytotoxicity and cellular uptake in non-tumor cells, murine fibroblast (L929) cells and tumor cells that overexpressed folate receptor (FR) β, and chronic myeloid leukemia cells in blast crisis (K562). Lastly, hemolysis assays were performed on human red blood cells. MNPsPMMA-FA presented an average mean diameter of 135 nm and a saturation magnetization (Ms) value of 37 emu/g of iron oxide, as well as superparamagnetic behavior. The MNPsPMMA-FA did not present cytotoxicity in L929 and K562 cells. Cellular uptake assays showed a higher uptake of MNPsPMMA-FA than MNPsPMMA in K562 cells when incubated at 37 °C. On the other hand, MNPsPMMA-FA showed a low uptake when endocytosis mechanisms were blocked at low temperature (4 °C), suggesting that the MNPsPMMA-FA uptake was mediated by endocytosis. High concentrations of MNPsPMMA-FA showed hemocompatibility when incubated for 24 h in human red blood cells. Therefore, our results suggest that these carrier systems can be an excellent alternative in targeted drug delivery via FR.

  16. Temperature dependent electronic conduction in semiconductors

    International Nuclear Information System (INIS)

    Roberts, G.G.; Munn, R.W.

    1980-01-01

    This review describes the temperature dependence of bulk-controlled electronic currents in semiconductors. The scope of the article is wide in that it contrasts conduction mechanisms in inorganic and organic solids and also single crystal and disordered semiconductors. In many experimental situations it is the metal-semiconductor contact or the interface between two dissimilar semiconductors that governs the temperature dependence of the conductivity. However, in order to keep the length of the review within reasonable bounds, these topics have been largely avoided and emphasis is therefore placed on bulk-limited currents. A central feature of electronic conduction in semiconductors is the concentrations of mobile electrons and holes that contribute to the conductivity. Various statistical approaches may be used to calculate these densities which are normally strongly temperature dependent. Section 1 emphasizes the relationship between the position of the Fermi level, the distribution of quantum states, the total number of electrons available and the absolute temperature of the system. The inclusion of experimental data for several materials is designed to assist the experimentalist in his interpretation of activation energy curves. Sections 2 and 3 refer to electronic conduction in disordered solids and molecular crystals, respectively. In these cases alternative approaches to the conventional band theory approach must be considered. For example, the velocities of the charge carriers are usually substantially lower than those in conventional inorganic single crystal semiconductors, thus introducing the possibility of an activated mobility. Some general electronic properties of these materials are given in the introduction to each of these sections and these help to set the conduction mechanisms in context. (orig.)

  17. Receptor-mediated endocytosis of lysozyme in renal proximal tubules of the frog Rana temporaria

    Directory of Open Access Journals (Sweden)

    E.V. Seliverstova

    2015-04-01

    Full Text Available The mechanism of protein reabsorption in the kidney of lower vertebrates remains insufficiently investigated in spite of raising interest to the amphibian and fish kidneys as a useful model for physiological and pathophysiological examinations. In the present study, we examined the renal tubular uptake and the internalization rote of lysozyme after its intravenous injection in the wintering frog Rana temporaria using immunohisto- and immunocytochemistry and specific markers for some endocytic compartments. The distinct expression of megalin and cubilin in the proximal tubule cells of lysozyme-injected frogs was revealed whereas kidney tissue of control animals showed no positive immunoreactivity. Lysozyme was detected in the apical endocytic compartment of the tubular cells and colocalized with clathrin 10 min after injection. After 20 min, lysozyme was located in the subapical compartment negative to clathrin (endosomes, and intracellular trafficking of lysozyme was coincided with the distribution of megalin and cubilin. However, internalized protein was retained in the endosomes and did not reach lysosomes within 30 min after treatment that may indicate the inhibition of intracellular trafficking in hibernating frogs. For the first time, we provided the evidence that lysozyme is filtered through the glomeruli and absorbed by receptor-mediated clathrin-dependent endocytosis in the frog proximal tubule cells. Thus, the protein uptake in the amphibian mesonephros is mediated by megalin and cubilin that confirms a critical role of endocytic receptors in the renal reabsorption of proteins in amphibians as in mammals.

  18. Theoretical analysis of time-dependent neutron spectra in bulk assemblies

    International Nuclear Information System (INIS)

    Akimoto, Tadashi; Ogawa, Yuichi; Togawa, Orihiko.

    1988-01-01

    Time-dependent neutron spectra in an iron assembly and in a graphite assembly are obtained with the one-dimensional S N calculation, in order an attempt to investigate the availability of these spectra to the benchmark test by the LINAC-TOF method for evaluation of nuclear data and numerical methods. The group constants are taken from the JAERI FAST SET Version 1, 2 and the ABBN SET. It was demonstrated by a sensitivity test that the time-dependent neutron spectra are sensitive to changes in the inelastic scattering cross section data in the iron assembly and to changes in the elastic scattering cross section data in the graphite assembly. Moreover, it is shown that the time-dependent spectra in the graphite assembly are sensitive to the group structure. Because some information about the neutron transport phenomena which has not been obtained in the stationary spectra is observed in the time-dependent spectra, the availability of the benchmark test based on the time-dependent spectra is indicated from the theoretical analysis. (author)

  19. High-Energy Laser Interaction with Gases, Droplets, and Bulk Liquids.

    Science.gov (United States)

    Jarzembski, Maurice Anthony

    Breakdown threshold intensities (I_ {rm TH}) were measured as functions of wavelengths and pressure for air, He, Ar, and Xe using a Nd:YAG pulsed laser. Multiphoton absorption dominates in the UV and cascade collision ionization dominates in the IR; however, both can be affected by other electron gain and loss processes. Presence of droplets lowers breakdown of gases due to field enhancements. Breakdown is initiated either in the droplet material or in the gas. At lambda = 0.532mum for a 50 μm dia. water droplet in He, Ar, and air for p pressure. For droplet -in-Xe, at p pressure. For droplet-in-Xe, at p 140 Torr, breakdown occurs outside the droplet and is dependent on gas pressure. Pressure dependence of breakdown was observed for 120mum dia. water droplets in Ar at p > 400 Torr. The required intensity for breakdown of droplet depends on I_{ rm TH} of bulk liquid and the effective field enhancement created by the droplet. The I _{rm TH} of droplet-in-air provides an upper limit to the propagation of a high energy laser beam in the atmosphere containing particles. By geometrical optics approach, a significant field enhancement located at the critical ring region, encircling the axis of the sphere in the forward direction at angle theta_{c}, was discovered where nonlinear processes can occur. This was confirmed experimentally and by Mie theory. Field enhancements calculated at the critical ring for water droplets of different sizes agree well with measurements. For a droplet of given size and real refractive index, the effective field enhancement and the volume over which it occurs are two important factors governing the occurrence of breakdown in droplets for both off resonance and on resonance conditions. Measurements of wavelength dependence of breakdown showed that in the UV, I_{rm TH} for droplets and bulk liquids were comparable and lower by few orders of magnitude from that of air. Transmittance and reflectance of bulk liquids in the UV change with

  20. Heterodimerization and endocytosis of Arabidopsis brassinosteroid receptors BRI1 and AtSERK3 (BAK1)

    DEFF Research Database (Denmark)

    Russinova, Eugenia; Borst, Jan-Willem; Kwaaitaal, Mark Adrianus Cornelis J

    2004-01-01

    us to localize each receptor independently in vivo. We show that BRI1, but not AtSERK3, homodimerizes in the plasma membrane, whereas BRI1 and AtSERK3 preferentially heterodimerize in the endosomes. Coexpression of BRI1 and AtSERK3 results in a change of the steady state distribution of both...... receptors because of accelerated endocytosis. Endocytic vesicles contain either BRI1 or AtSERK3 alone or both. We propose that the AtSERK3 protein is involved in changing the equilibrium between plasma membrane-located BRI1 homodimers and endocytosed BRI1-AtSERK3 heterodimers....

  1. AC characterization of bulk organic solar cell in the dark and under illumination

    International Nuclear Information System (INIS)

    Váry, Michal; Perný, Milan; Šály, Vladimír; Packa, Juraj

    2014-01-01

    Highlights: • A study of organic bulk photovoltaic (PV) solar cell. • Current–voltage characteristics in the dark and under illumination. • AC measurements, both under illumination and in the dark conditions. • Equivalent AC circuit. • Effective lifetime assigned with electron–hole recombination and diffusion time of the electron was estimated. - Abstract: Impedance spectroscopy has been used widely to evaluate the transport processes in photovoltaic, mainly based on inorganic semiconductors, structures – solar cells. The aim of this research was to characterize improved organic bulk photovoltaic (PV) solar cells exploiting this method. Progress in technology of investigated organic solar cell involves the use of an active layer based on low band gap type of polymer. The organic PV cell with front transparent electrode and rear metal electrode and active layer produced by Konarka Technologies was analyzed by electrical DC and AC measurements. Current–voltage (I–V) characteristics in the dark and under illumination were measured and basic PV parameters were calculated. AC measurements, both under illumination and in the dark conditions, were processed in order to identify electronic behavior using equivalent AC circuit which was suggested by fitting of measured impedance data. Circuit with the best correlation to measured data is analyzed in details. Voltage and frequency dependences of fitted equivalent circuit components and calculated parameters are explained and presented in the paper

  2. Mechanical improvement of metal reinforcement rings for a finite ring-shaped superconducting bulk

    Science.gov (United States)

    Huang, Chen-Guang; Zhou, You-He

    2018-03-01

    As a key technique, reinforcement of type-II superconducting bulks with metal rings can efficiently improve their mechanical properties to enhance the maximum trapped field. In this paper, we study the magnetostrictive and fracture behaviors of a finite superconducting ring bulk reinforced by three typical reinforcing structures composed of metal rings during the magnetizing process by means of the minimization of magnetic energy and the finite element method. After a field-dependent critical current density is adopted, the magnetostriction, pinning-induced stress, and crack tip stress intensity factor are calculated considering the demagnetization effects. The results show that the mechanical properties of the ring bulk are strongly dependent on the reinforcing structure and the material and geometrical parameters of the metal rings. Introducing the metal ring can significantly reduce the hoop stress, and the reduction effect by internal reinforcement is much improved relative to external reinforcement. By comparison, bilateral reinforcement seems to be the best candidate structure. Only when the metal rings have particular Young's modulus and radial thickness will they contribute to improve the mechanical properties the most. In addition, if an edge crack is pre-existing in the ring bulk, the presence of metal rings can effectively avoid crack propagation since it reduces the crack tip stress intensity factor by nearly one order of magnitude.

  3. Tyrosine 769 of the keratinocyte growth factor receptor is required for receptor signaling but not endocytosis

    International Nuclear Information System (INIS)

    Ceridono, Mara; Belleudi, Francesca; Ceccarelli, Simona; Torrisi, Maria Rosaria

    2005-01-01

    Keratinocyte growth factor receptor (KGFR) is a receptor tyrosine kinase expressed on epithelial cells which belongs to the family of fibroblast growth factor receptors (FGFRs). Following ligand binding, KGFR is rapidly autophosphorylated on specific tyrosine residues in the intracellular domain, recruits substrate proteins, and is rapidly internalized by clathrin-mediated endocytosis. The role of different autophosphorylation sites in FGFRs, and in particular the role of the tyrosine 766 in FGFR1, first identified as PLCγ binding site, has been extensively studied. We analyzed here the possible role of the tyrosine 769 in KGFR, corresponding to tyrosine 766 in FGFR1, in the regulation of KGFR signal transduction and MAPK activation as well as in the control of the endocytic process of KGFR. A mutant KGFR in which tyrosine 769 was substituted by phenylalanine was generated and transfected in NIH3T3 and HeLa cells. Our results indicate that tyrosine 769 is required for the binding to KGFR and tyrosine phosphorylation of PLCγ as well as for the full activation of MAPKs and for cell proliferation through the regulation of FRS2 tyrosine phosphorylation, suggesting that this residue represents a key regulator of KGFR signal transduction. Our data also show that tyrosine 769 is not involved in the regulation of the endocytic process of KGFR

  4. Bulk Genotyping of Biopsies Can Create Spurious Evidence for Hetereogeneity in Mutation Content.

    Directory of Open Access Journals (Sweden)

    Rumen Kostadinov

    2016-04-01

    Full Text Available When multiple samples are taken from the neoplastic tissues of a single patient, it is natural to compare their mutation content. This is often done by bulk genotyping of whole biopsies, but the chance that a mutation will be detected in bulk genotyping depends on its local frequency in the sample. When the underlying mutation count per cell is equal, homogenous biopsies will have more high-frequency mutations, and thus more detectable mutations, than heterogeneous ones. Using simulations, we show that bulk genotyping of data simulated under a neutral model of somatic evolution generates strong spurious evidence for non-neutrality, because the pattern of tissue growth systematically generates differences in biopsy heterogeneity. Any experiment which compares mutation content across bulk-genotyped biopsies may therefore suggest mutation rate or selection intensity variation even when these forces are absent. We discuss computational and experimental approaches for resolving this problem.

  5. Bulk Genotyping of Biopsies Can Create Spurious Evidence for Hetereogeneity in Mutation Content.

    Science.gov (United States)

    Kostadinov, Rumen; Maley, Carlo C; Kuhner, Mary K

    2016-04-01

    When multiple samples are taken from the neoplastic tissues of a single patient, it is natural to compare their mutation content. This is often done by bulk genotyping of whole biopsies, but the chance that a mutation will be detected in bulk genotyping depends on its local frequency in the sample. When the underlying mutation count per cell is equal, homogenous biopsies will have more high-frequency mutations, and thus more detectable mutations, than heterogeneous ones. Using simulations, we show that bulk genotyping of data simulated under a neutral model of somatic evolution generates strong spurious evidence for non-neutrality, because the pattern of tissue growth systematically generates differences in biopsy heterogeneity. Any experiment which compares mutation content across bulk-genotyped biopsies may therefore suggest mutation rate or selection intensity variation even when these forces are absent. We discuss computational and experimental approaches for resolving this problem.

  6. Irreversibility line and magnetic field dependence of the critical current in superconducting MgB sub 2 bulk samples

    CERN Document Server

    Gioacchino, D D; Tripodi, P; Grimaldi, G

    2003-01-01

    The third harmonic components of the ac susceptibility of MgB sub 2 bulk samples have been measured as a function of applied magnetic fields, together with standard magnetization cycles. The irreversibility line (IL) of the magnetic field has been extracted from the onset of the third harmonic components. Using a (1 - t) supalpha glass/liquid best fit where alpha 1.27 IL shows a coherent length xi divergence with exponent nu = 0.63, which indicates a 3D behaviour. Moreover, using the numerical solution of the non-linear magnetic diffusion equation, considering the creep model in a 3D vortex glass, a good description of the vortex dynamics has been obtained. The behaviour of the magnetization amplitude (approx Hz) and the ac susceptibility signals (kHz), at different applied magnetic fields, 3.5 T < H sub d sub c < 4.5 T, and at the reduced temperature 0.86 < t < 0.93 (T = 22 K), shows that the superconducting dynamic response of vortices in the MgB sub 2 samples is not evidently dependent on the f...

  7. Bulk analysis using nuclear techniques

    International Nuclear Information System (INIS)

    Borsaru, M.; Holmes, R.J.; Mathew, P.J.

    1983-01-01

    Bulk analysis techniques developed for the mining industry are reviewed. Using penetrating neutron and #betta#-radiations, measurements are obtained directly from a large volume of sample (3-30 kg) #betta#-techniques were used to determine the grade of iron ore and to detect shale on conveyor belts. Thermal neutron irradiation was developed for the simultaneous determination of iron and aluminium in iron ore on a conveyor belt. Thermal-neutron activation analysis includes the determination of alumina in bauxite, and manganese and alumina in manganese ore. Fast neutron activation analysis is used to determine silicon in iron ores, and alumina and silica in bauxite. Fast and thermal neutron activation has been used to determine the soil in shredded sugar cane. (U.K.)

  8. Puerarin activates endothelial nitric oxide synthase through estrogen receptor-dependent PI3-kinase and calcium-dependent AMP-activated protein kinase

    Energy Technology Data Exchange (ETDEWEB)

    Hwang, Yong Pil; Kim, Hyung Gyun [Department of Toxicology, College of Pharmacy, Chungnam National University, Daejeon (Korea, Republic of); Hien, Tran Thi [College of Pharmacy, Chosun University, Gwangju (Korea, Republic of); Jeong, Myung Ho [Heart Research Center, Chonnam National University Hospital, Gwangju (Korea, Republic of); Jeong, Tae Cheon, E-mail: taecheon@ynu.ac.kr [College of Pharmacy, Yeungnam University, Gyungsan (Korea, Republic of); Jeong, Hye Gwang, E-mail: hgjeong@cnu.ac.kr [Department of Toxicology, College of Pharmacy, Chungnam National University, Daejeon (Korea, Republic of)

    2011-11-15

    The cardioprotective properties of puerarin, a natural product, have been attributed to the endothelial nitric oxide synthase (eNOS)-mediated production of nitric oxide (NO) in EA.hy926 endothelial cells. However, the mechanism by which puerarin activates eNOS remains unclear. In this study, we sought to identify the intracellular pathways underlying eNOS activation by puerarin. Puerarin induced the activating phosphorylation of eNOS on Ser1177 and the production of NO in EA.hy926 cells. Puerarin-induced eNOS phosphorylation required estrogen receptor (ER)-mediated phosphatidylinositol 3-kinase (PI3K)/Akt signaling and was reversed by AMP-activated protein kinase (AMPK) and calcium/calmodulin-dependent kinase II (CaMKII) inhibition. Importantly, puerarin inhibited the adhesion of tumor necrosis factor (TNF)-{alpha}-stimulated monocytes to endothelial cells and suppressed the TNF-{alpha} induced expression of intercellular cell adhesion molecule-1. Puerarin also inhibited the TNF-{alpha}-induced nuclear factor-{kappa}B activation, which was attenuated by pretreatment with N{sup G}-nitro-L-arginine methyl ester, a NOS inhibitor. These results indicate that puerarin stimulates eNOS phosphorylation and NO production via activation of an estrogen receptor-mediated PI3K/Akt- and CaMKII/AMPK-dependent pathway. Puerarin may be useful for the treatment or prevention of endothelial dysfunction associated with diabetes and cardiovascular disease. -- Highlights: Black-Right-Pointing-Pointer Puerarin induced the phosphorylation of eNOS and the production of NO. Black-Right-Pointing-Pointer Puerarin activated eNOS through ER-dependent PI3-kinase and Ca{sup 2+}-dependent AMPK. Black-Right-Pointing-Pointer Puerarin-induced NO was involved in the inhibition of NF-kB activation. Black-Right-Pointing-Pointer Puerarin may help for prevention of vascular dysfunction and diabetes.

  9. Ferromagnetic transitions of a spin-one Ising film in a surface and bulk transverse fields

    International Nuclear Information System (INIS)

    Saber, A.; Lo Russo, S.; Mattei, G.; Mattoni, A.

    2002-01-01

    Using the effective field theory method, we have calculated the Curie temperature of a spin-one Ising ferromagnetic film in a surface and bulk transverse fields. Numerical calculations give phase diagrams under various parameters. Surface exchange enhancement is considered. The dependence of the critical transverse field on film thickness, and phase diagrams in the fields, critical surface transverse field versus the bulk one are presented

  10. Activating antioxidant enzymes, hyoscyamine and scopolamine biosynthesis of Hyoscyamus niger L. plants with nano-sized titanium dioxide and bulk application

    Directory of Open Access Journals (Sweden)

    Mansour GHORBANPOUR

    2015-11-01

    Full Text Available  Application of nanotechnology is now widely distributed overall the life, especially in agricultural systems. This study intended to indicate the impacts of nano-sized titanium dioxide particles (NT and bulk (BT on antioxidant enzymes activities including superoxide dismutase (SOD, peroxidase (POX and catalase (CAT, and variations of two major tropane alkaloids such as hyoscyamine (HYO and scopolamine (SCO in Hyoscyamus niger L. Plants were treated with different concentrations of NT and BT (0, 20, 40 and 80 mg l-1. Alkaloids extracted were identified by gas chromatography (GC and gas chromatography-mass spectrometry (GC-MS analysis. Results showed that SOD activity increased with increasing titanium dioxide concentration in both nano-particles and bulk treated plants. However, the highest and the lowest POX activity were observed in plants exposed to NT at 40 mg l-1 and control, respectively. Generally, all tested enzymes activities were higher in NT treated plants that those of BT except CAT activity at 80 mg l-1. The highest alkaloids content values, HYO: 0.286 g kg-1 and SCO: 0.126 g kg-1, were achieved in plants treated with NT at 80 and 20 mg l-1, respectively. The maximum and minimum plant biomass and subsequently total alkaloids yield were obtained in plants exposed to NT at 40 mg l-1 and controls, respectively. Our results suggest that NT in appropriate level (40 mg l-1 may act as an elicitor for biochemical responses and tropane alkaloids biosynthesis in H. niger plants. 

  11. Passion and dependency in online shopping activities.

    Science.gov (United States)

    Wang, Chih-Chien; Yang, Hui-Wen

    2007-04-01

    This study examines the influence of harmonious passion (HP) and obsessive passion (OP) to online shopping dependency. The results show that both HP and OP might lead to online shopping dependency and online shoppers with OP are more dependent on online shopping activities. In addition, this study also found out that HP and OP could be denoted as a sequence of different intensities of passion, where HP might be a necessity of OP.

  12. Effects of effluent organic matter characteristics on the removal of bulk organic matter and selected pharmaceutically active compounds during managed aquifer recharge: Column study

    KAUST Repository

    Maeng, Sungkyu; Sharma, Saroj K.; Abel, Chol D T; Magic-Knezev, Aleksandra; Song, Kyungguen; Amy, Gary L.

    2012-01-01

    Soil column experiments were conducted to investigate the effects of effluent organic matter (EfOM) characteristics on the removal of bulk organic matter (OM) and pharmaceutically active compounds (PhACs) during managed aquifer recharge (MAR

  13. Ligand-Occupied Integrin Internalization Links Nutrient Signaling to Invasive Migration

    Directory of Open Access Journals (Sweden)

    Elena Rainero

    2015-01-01

    Full Text Available Integrin trafficking is key to cell migration, but little is known about the spatiotemporal organization of integrin endocytosis. Here, we show that α5β1 integrin undergoes tensin-dependent centripetal movement from the cell periphery to populate adhesions located under the nucleus. From here, ligand-engaged α5β1 integrins are internalized under control of the Arf subfamily GTPase, Arf4, and are trafficked to nearby late endosomes/lysosomes. Suppression of centripetal movement or Arf4-dependent endocytosis disrupts flow of ligand-bound integrins to late endosomes/lysosomes and their degradation within this compartment. Arf4-dependent integrin internalization is required for proper lysosome positioning and for recruitment and activation of mTOR at this cellular subcompartment. Furthermore, nutrient depletion promotes subnuclear accumulation and endocytosis of ligand-engaged α5β1 integrins via inhibition of mTORC1. This two-way regulatory interaction between mTORC1 and integrin trafficking in combination with data describing a role for tensin in invasive cell migration indicate interesting links between nutrient signaling and metastasis.

  14. Light-dependent electrogenic activity of cyanobacteria.

    Directory of Open Access Journals (Sweden)

    John M Pisciotta

    2010-05-01

    Full Text Available Cyanobacteria account for 20-30% of Earth's primary photosynthetic productivity and convert solar energy into biomass-stored chemical energy at the rate of approximately 450 TW [1]. These single-cell microorganisms are resilient predecessors of all higher oxygenic phototrophs and can be found in self-sustaining, nitrogen-fixing communities the world over, from Antarctic glaciers to the Sahara desert [2].Here we show that diverse genera of cyanobacteria including biofilm-forming and pelagic strains have a conserved light-dependent electrogenic activity, i.e. the ability to transfer electrons to their surroundings in response to illumination. Naturally-growing biofilm-forming photosynthetic consortia also displayed light-dependent electrogenic activity, demonstrating that this phenomenon is not limited to individual cultures. Treatment with site-specific inhibitors revealed the electrons originate at the photosynthetic electron transfer chain (P-ETC. Moreover, electrogenic activity was observed upon illumination only with blue or red but not green light confirming that P-ETC is the source of electrons. The yield of electrons harvested by extracellular electron acceptor to photons available for photosynthesis ranged from 0.05% to 0.3%, although the efficiency of electron harvesting likely varies depending on terminal electron acceptor.The current study illustrates that cyanobacterial electrogenic activity is an important microbiological conduit of solar energy into the biosphere. The mechanism responsible for electrogenic activity in cyanobacteria appears to be fundamentally different from the one exploited in previously discovered electrogenic bacteria, such as Geobacter, where electrons are derived from oxidation of organic compounds and transported via a respiratory electron transfer chain (R-ETC [3], [4]. The electrogenic pathway of cyanobacteria might be exploited to develop light-sensitive devices or future technologies that convert solar

  15. Prenatal and adult androgen activities in alcohol dependence.

    Science.gov (United States)

    Lenz, B; Mühle, C; Braun, B; Weinland, C; Bouna-Pyrrou, P; Behrens, J; Kubis, S; Mikolaiczik, K; Muschler, M-R; Saigali, S; Sibach, M; Tanovska, P; Huber, S E; Hoppe, U; Eichler, A; Heinrich, H; Moll, G H; Engel, A; Goecke, T W; Beckmann, M W; Fasching, P A; Müller, C P; Kornhuber, J

    2017-07-01

    Alcohol dependence is more prevalent in men than in women. The evidence for how prenatal and adult androgens influence alcohol dependence is limited. We investigated the effects of prenatal and adult androgen activity on alcohol dependence. Moreover, we studied how the behaviours of pregnant women affect their children's prenatal androgen load. We quantified prenatal androgen markers (e.g., second-to-fourth finger length ratio [2D : 4D]) and blood androgens in 200 early-abstinent alcohol-dependent in-patients and 240 controls (2013-2015, including a 12-month follow-up). We also surveyed 134 women during pregnancy (2005-2007) and measured the 2D : 4D of their children (2013-2016). The prenatal androgen loads were higher in the male alcohol-dependent patients compared to the controls (lower 2D : 4D, P = 0.004) and correlated positively with the patients' liver transaminase activities (P alcohol withdrawal severity (P = 0.019). Higher prenatal androgen loads and increasing androgen levels during withdrawal predicted earlier and more frequent 12-month hospital readmission in alcohol-dependent patients (P alcohol (P = 0.010) and tobacco consumption (P = 0.017), and lifetime stressors (P = 0.019) of women during pregnancy related positively to their children's prenatal androgen loads (lower 2D : 4D). Androgen activities in alcohol-dependent patients and behaviours of pregnant women represent novel preventive and therapeutic targets of alcohol dependence. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Different cAMP sources are critically involved in G protein-coupled receptor CRHR1 signaling.

    Science.gov (United States)

    Inda, Carolina; Dos Santos Claro, Paula A; Bonfiglio, Juan J; Senin, Sergio A; Maccarrone, Giuseppina; Turck, Christoph W; Silberstein, Susana

    2016-07-18

    Corticotropin-releasing hormone receptor 1 (CRHR1) activates G protein-dependent and internalization-dependent signaling mechanisms. Here, we report that the cyclic AMP (cAMP) response of CRHR1 in physiologically relevant scenarios engages separate cAMP sources, involving the atypical soluble adenylyl cyclase (sAC) in addition to transmembrane adenylyl cyclases (tmACs). cAMP produced by tmACs and sAC is required for the acute phase of extracellular signal regulated kinase 1/2 activation triggered by CRH-stimulated CRHR1, but only sAC activity is essential for the sustained internalization-dependent phase. Thus, different cAMP sources are involved in different signaling mechanisms. Examination of the cAMP response revealed that CRH-activated CRHR1 generates cAMP after endocytosis. Characterizing CRHR1 signaling uncovered a specific link between CRH-activated CRHR1, sAC, and endosome-based signaling. We provide evidence of sAC being involved in an endocytosis-dependent cAMP response, strengthening the emerging model of GPCR signaling in which the cAMP response does not occur exclusively at the plasma membrane and introducing the notion of sAC as an alternative source of cAMP. © 2016 Inda et al.

  17. Sandwich-cell-type bulk-heterojunction organic solar cells utilizing liquid crystalline phthalocyanine

    Science.gov (United States)

    Nakata, Yuya; Usui, Toshiki; Nishikawa, Yuki; Nekelson, Fabien; Shimizu, Yo; Fujii, Akihiko; Ozaki, Masanori

    2018-03-01

    Sandwich-cell-type bulk-heterojunction organic solar cells utilizing the liquid crystalline phthalocyanine, 1,4,8,11,15,18,22,25-octahexylphthalocyanine (C6PcH2), have been fabricated and their photovoltaic properties have been studied. The short-circuit current (J SC) and power conversion efficiency (PCE) depended on the blend ratio of donor and acceptor molecules, and the maximum performance, such as J SC of 3.4 mA/cm2 and PCE of 0.67%, was demonstrated, when the blend ratio of the acceptor was 10 mol %. The photovoltaic properties were discussed by taking the relationship between the column axis direction of C6PcH2 and the carrier mobility in the active layer into consideration.

  18. Microhardness of bulk-fill composite materials

    OpenAIRE

    Kelić, Katarina; Matić, Sanja; Marović, Danijela; Klarić, Eva; Tarle, Zrinka

    2016-01-01

    The aim of the study was to determine microhardness of high- and low-viscosity bulk-fill composite resins and compare it with conventional composite materials. Four materials of high-viscosity were tested, including three bulk-fills: QuiXfi l (QF), x-tra fil (XTF) and Tetric EvoCeram Bulk Fill (TEBCF), while nanohybrid composite GrandioSO (GSO) served as control. The other four were low-viscosity composites, three bulk-fill materials: Smart Dentin Replacement (SDR), Venus Bulk Fill (VBF) and ...

  19. Receptor-mediated endocytosis of polypeptide hormones is a regulated process: inhibition of [125I]iodoinsulin internalization in hypoinsulinemic diabetes of rat and man

    International Nuclear Information System (INIS)

    Carpentier, J.L.; Robert, A.; Grunberger, G.; van Obberghen, E.; Freychet, P.; Orci, L.; Gorden, P.

    1986-01-01

    Much data suggest that receptor-mediated endocytosis is regulated in states of hormone excess. Thus, in hyperinsulinemic states there is an accelerated loss of cell surface insulin receptors. In the present experiments we addressed this question in hypoinsulinemic states, in which insulin binding to cell surface receptors is generally increased. In hepatocytes obtained from hypoinsulinemic streptozotocin-induced diabetic rats, [ 125 I]iodoglucagon internalization was increased, while at the same time [ 125 I]iodoinsulin internalization was decreased. The defect in [ 125 I]iodoinsulin internalization was corrected by insulin treatment of the animal. In peripheral blood monocytes from patients with type I insulinopenic diabetes, internalization of [ 125 I]iodoinsulin was impaired; this defect was not present in insulin-treated patients. These data in the hypoinsulinemic rat and human diabetes suggest that receptor-mediated endocytosis is regulated in states of insulin deficiency as well as insulin excess. Delayed or reduced internalization of the insulin-receptor complex could amplify the muted signal caused by deficient hormone secretion

  20. Comparisons of Soil Properties, Enzyme Activities and Microbial Communities in Heavy Metal Contaminated Bulk and Rhizosphere Soils of Robinia pseudoacacia L. in the Northern Foot of Qinling Mountain

    Directory of Open Access Journals (Sweden)

    Yurong Yang

    2017-11-01

    Full Text Available The toxic effects of heavy metal (HM contamination on plant metabolism and soil microorganisms have been emphasized recently; however, little is known about the differences in soil physical, chemical, and biological properties between bulk and rhizosphere soils contaminated with HMs in forest ecosystem. The present study was conducted to evaluate the rhizosphere effect on soil properties, enzyme activities and bacterial communities associated with Robinia pseudoacacia L. along a HM contamination gradient. Soil organic matter (SOM, available nitrogen (AN and phosphorus (AP contents were significantly higher in rhizosphere soil than those in bulk soil at HM contaminated sites (p < 0.05. Compared to bulk soil, activities of four soil enzymes indicative of C cycle (β-glucosidase, N cycle (protease, urease and P cycle (alkaline phosphatase in rhizosphere soil across all study sites increased by 47.5%, 64.1%, 52.9% and 103.8%, respectively. Quantitative PCR (qPCR and restriction fragment length polymorphism (RFLP were used to determine the relative abundance, composition and diversity of bacteria in both bulk and rhizosphere soils, respectively. The copy number of bacterial 16S rRNA gene in bulk soil was significantly lower than that in rhizosphere soil (p < 0.05, and it had significantly negative correlations with total/DTPA-extractable Pb concentrations (p < 0.01. Alphaproteobacteria, Gammaproteobacteria and Firmicutes were the most dominant groups of bacteria at different study sites. The bacterial diversity index of Species richness (S and Margalef (dMa were significantly higher in rhizosphere soil compared with those in bulk soil, although no difference could be found in Simpson index (D between bulk and rhizosphere soils (p > 0.05. Redundancy analysis (RDA results showed that soil pH, EC, SOM and total/DTPA-extractable Pb concentrations were the most important variables affecting relative abundance, composition and diversity of bacteria (p < 0

  1. High-temperature bulk acoustic wave sensors

    International Nuclear Information System (INIS)

    Fritze, Holger

    2011-01-01

    Piezoelectric crystals like langasite (La 3 Ga 5 SiO 14 , LGS) and gallium orthophosphate (GaPO 4 ) exhibit piezoelectrically excited bulk acoustic waves at temperatures of up to at least 1450 °C and 900 °C, respectively. Consequently, resonant sensors based on those materials enable new sensing approaches. Thereby, resonant high-temperature microbalances are of particular interest. They correlate very small mass changes during film deposition onto resonators or gas composition-dependent stoichiometry changes of thin films already deposited onto the resonators with the resonance frequency shift of such devices. Consequently, the objective of the work is to review the high-temperature properties, the operation limits and the measurement principles of such resonators. The electromechanical properties of high-temperature bulk acoustic wave resonators such as mechanical stiffness, piezoelectric and dielectric constant, effective viscosity and electrical conductivity are described using a one-dimensional physical model and determined accurately up to temperatures as close as possible to their ultimate limit. Insights from defect chemical models are correlated with the electromechanical properties of the resonators. Thereby, crucial properties for stable operation as a sensor under harsh conditions are identified to be the formation of oxygen vacancies and the bulk conductivity. Operation limits concerning temperature, oxygen partial pressure and water vapor pressure are given. Further, application-relevant aspects such as temperature coefficients, temperature compensation and mass sensitivity are evaluated. In addition, approximations are introduced which make the exact model handy for routine data evaluation. An equivalent electrical circuit for high-temperature resonator devices is derived based on the one-dimensional physical model. Low- and high-temperature approximations are introduced. Thereby, the structure of the equivalent circuit corresponds to the

  2. High-temperature bulk acoustic wave sensors

    Science.gov (United States)

    Fritze, Holger

    2011-01-01

    Piezoelectric crystals like langasite (La3Ga5SiO14, LGS) and gallium orthophosphate (GaPO4) exhibit piezoelectrically excited bulk acoustic waves at temperatures of up to at least 1450 °C and 900 °C, respectively. Consequently, resonant sensors based on those materials enable new sensing approaches. Thereby, resonant high-temperature microbalances are of particular interest. They correlate very small mass changes during film deposition onto resonators or gas composition-dependent stoichiometry changes of thin films already deposited onto the resonators with the resonance frequency shift of such devices. Consequently, the objective of the work is to review the high-temperature properties, the operation limits and the measurement principles of such resonators. The electromechanical properties of high-temperature bulk acoustic wave resonators such as mechanical stiffness, piezoelectric and dielectric constant, effective viscosity and electrical conductivity are described using a one-dimensional physical model and determined accurately up to temperatures as close as possible to their ultimate limit. Insights from defect chemical models are correlated with the electromechanical properties of the resonators. Thereby, crucial properties for stable operation as a sensor under harsh conditions are identified to be the formation of oxygen vacancies and the bulk conductivity. Operation limits concerning temperature, oxygen partial pressure and water vapor pressure are given. Further, application-relevant aspects such as temperature coefficients, temperature compensation and mass sensitivity are evaluated. In addition, approximations are introduced which make the exact model handy for routine data evaluation. An equivalent electrical circuit for high-temperature resonator devices is derived based on the one-dimensional physical model. Low- and high-temperature approximations are introduced. Thereby, the structure of the equivalent circuit corresponds to the Butterworth

  3. Scaling laws in high-energy inverse compton scattering. II. Effect of bulk motions

    International Nuclear Information System (INIS)

    Nozawa, Satoshi; Kohyama, Yasuharu; Itoh, Naoki

    2010-01-01

    We study the inverse Compton scattering of the CMB photons off high-energy nonthermal electrons. We extend the formalism obtained by the previous paper to the case where the electrons have nonzero bulk motions with respect to the CMB frame. Assuming the power-law electron distribution, we find the same scaling law for the probability distribution function P 1,K (s) as P 1 (s) which corresponds to the zero bulk motions, where the peak height and peak position depend only on the power-index parameter. We solved the rate equation analytically. It is found that the spectral intensity function also has the same scaling law. The effect of the bulk motions to the spectral intensity function is found to be small. The present study will be applicable to the analysis of the x-ray and gamma-ray emission models from various astrophysical objects with nonzero bulk motions such as radio galaxies and astrophysical jets.

  4. DESIGN OF THE DEMOSNTRATION BULK VITRIFICATION SYSTEM FOR THE SUPPLEMENTAL TREATMENT OF LOW ACTIVITY TANK WASTE AT HANFORD

    International Nuclear Information System (INIS)

    VAN BEEK JE

    2008-01-01

    In June 2004, the Demonstration Bulk Vitrification System (DBVS) was initiated with the intent to design, construct, and operate a full-scale bulk vitrification pilot-plant to treat low-activity tank waste from Hanford Tank 241-S-109. The DBVS facility uses In-Container Vitrification(trademark) (ICV(trademark)) at the core of the treatment process. The basic process steps combine liquid low-activity waste (LAW) and glassformers; dry the mixture; and then vitrify the mixture in a batch feed-while-melt process in a refractory lined steel container. Off-gases are processed through a state-of-the-art air pollution control system including sintered-metal filtration, thermal oxidation, acid gas scrubbing, and high-efficiency particulate air (HEPA) and high-efficiency gas adsorber (HEGA) filtration. Testing has focused on development and validation of the waste dryer, ICV, and sintered-metal filters (SMFs) equipment, operations enhancements, and glass formulation. With a parallel testing and design process, testing has allowed improvements to the DBVS equipment configuration and operating methodology, since its original inception. Design improvements include optimization of refractory panels in the ICV, simplifying glassformer addition equipment, increasing the number of waste feed chutes to the ICV, and adding capability for remote clean-out of piping, In addition, the U.S. Department of Energy (DOE) has provided an independent review of the entire DBVS process. While the review did not find any fatal flaws, some technical issues were identified that required a re-evaluation of the DBVS design and subsequent changes to the design. A 100 percent design package for the pilot plant will be completed and submitted to DOE for review in early 2008 that incorporates process improvements substantiated through testing and reviews. This paper provides a description of the bulk vitrification process and a discussion of major equipment design changes that have occurred based on full

  5. Surface-spin magnetism of antiferromagnetic NiO in nanoparticle and bulk morphology

    International Nuclear Information System (INIS)

    Jagodic, M; Jaglicic, Z; Jelen, A; Dolinsek, J; Lee, Jin Bae; Kim, Hae Jin; Kim, Young-Min

    2009-01-01

    The surface-spin magnetism of the antiferromagnetic (AFM) material NiO in nanoparticle and bulk morphology was investigated by magnetic measurements (temperature-dependent zero-field-cooled (zfc) and field-cooled (fc) dc susceptibility, ac susceptibility and zfc and fc hysteresis loops). We addressed the question of whether the multisublattice ordering of the uncompensated surface spins and the exchange bias (EB) effect are only present in the nanoparticles, originating from their high surface-to-volume ratio or if these surface phenomena are generally present in the AFM materials regardless of their bulky or nanoparticle morphology, but the effect is just too small to be detected experimentally in the bulk due to a very small surface magnetization. Performing experiments on the NiO nanoparticles of different sizes and bulk NiO grains, we show that coercivity enhancement and hysteresis loop shift in the fc experiments, considered to be the key experimental manifestations of multisublattice ordering and the EB effect, are true nanoscale phenomena only present in the nanoparticles and absent in the bulk.

  6. Bulk fill restoratives: to cap or not to cap--that is the question?

    Science.gov (United States)

    Tomaszewska, Iwona M; Kearns, Jennifer O; Ilie, Nicoleta; Fleming, Garry J P

    2015-03-01

    To assess the cuspal deflection and cervical microleakage scores of standardised large mesio-occlusal-distal (MOD) cavities filled with different restoration protocols: (1) conventional resin restoratives, (2) bulk fill flowable base materials 'capped' with a conventional dimethacrylate resin-based composite (RBC) or (3) bulk fill resin restorative materials. Standardised MOD cavities were prepared in sixty-four sound maxillary premolar teeth and randomly allocated to eight groups. Restorations were placed in conjunction with a universal bonding system and resin restorative materials were irradiated with a quartz-tungsten-halogen light-curing-unit. Restoration protocol (eight oblique increments of conventional resin restorative, bulk fill flowable base and two occlusal 'capping' RBC increments (three increments in total) or bulk fill resin restorative (two increments)) was the dependent variable. A twin channel deflection measuring gauge measured the buccal and palatal cuspal deflections. Teeth were thermally fatigued, immersed in a 0.2% basic fuchsin dye for 24h, sectioned and examined for cervical microleakage score. Post hoc Tukey's tests highlighted significant differences in the mean total cuspal deflection values between resin restoratives (p fill flowable base materials with occlusal 'capping' RBC increments (restoration protocol 2) compared with bulk fill resin restoratives (restoration protocol 3). Not all bulk fill flowable materials or bulk fill resin restoratives behave in a similar fashion when used to restore standardised MOD cavities in maxillary premolar teeth and material selection is vital in the absence of clinical data. Poorly performing bulk fill flowable materials or bulk fill restoratives can be identified using the cuspal deflection and cervical microleakage protocol which could save the complications encountered clinically when restoring Class II restorations. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Evaluation of Connexin 43 Redistribution and Endocytosis in Astrocytes Subjected to Ischemia/Reperfusion or Oxygen-Glucose Deprivation and Reoxygenation

    Directory of Open Access Journals (Sweden)

    Hongyan Xie

    2017-01-01

    Full Text Available Connexin 43 (Cx43 is the major component protein in astrocytic gap junction communication. Recent studies have shown the cellular processes of gap junction internalization and degradation, but many details remain unknown. This study investigated the distribution of Cx43 and its mechanism after ischemic insult. Astrocyte culture system and a model of ischemia/reperfusion (IR or oxygen-glucose deprivation and reoxygenation (OGDR were established. Cx43 distribution was observed by laser scanning confocal microscopy under different cultivation conditions. Western blot and RT-PCR assays were applied to quantify Cx43 and MAPRE1 (microtubule-associated protein RP/EB family member 1 expression at different time points. The total number of Cx43 was unchanged in the normal and IR/OGDR groups, but Cx43 particles in the cytoplasm of the IR/OGDR group were significantly greater than that of the normal group. Particles in the cytoplasm were significantly fewer after endocytosis was blocked by dynasore. There was no difference among the groups at each time point regarding protein or gene expression of MAPRE1. We concluded that internalization of Cx43 into the cytoplasm occurred during ischemia, which was partially mediated through endocytosis, not by the change of Cx43 quantity. Moreover, internalization was not related to microtubule transport.

  8. Field dependence-independence and participation in physical activity by college students.

    Science.gov (United States)

    Liu, Wenhao

    2006-06-01

    Field-independent individuals, compared with field-dependent individuals, have higher sports potential and advantages in sport-related settings. Little research, however, has been conducted on the association of field dependence-independence and participation in physical activity. The study examined this association for college students who participated in physical activities in and beyond physical education classes. The Group Embedded Figures Test distinguished 40 field-dependent from 40 field-independent participants. Activity logs during one semester showed that field-independent participants were significantly more physically active and their physical activity behaviors were more sport-related than those of field-dependent participants.

  9. Suppression of CHRN endocytosis by carbonic anhydrase CAR3 in the pathogenesis of myasthenia gravis.

    Science.gov (United States)

    Du, Ailian; Huang, Shiqian; Zhao, Xiaonan; Feng, Kuan; Zhang, Shuangyan; Huang, Jiefang; Miao, Xiang; Baggi, Fulvio; Ostrom, Rennolds S; Zhang, Yanyun; Chen, Xiangjun; Xu, Congfeng

    2017-01-01

    Myasthenia gravis is an autoimmune disorder of the neuromuscular junction manifested as fatigable muscle weakness, which is typically caused by pathogenic autoantibodies against postsynaptic CHRN/AChR (cholinergic receptor nicotinic) in the endplate of skeletal muscle. Our previous studies have identified CA3 (carbonic anhydrase 3) as a specific protein insufficient in skeletal muscle from myasthenia gravis patients. In this study, we investigated the underlying mechanism of how CA3 insufficiency might contribute to myasthenia gravis. Using an experimental autoimmune myasthenia gravis animal model and the skeletal muscle cell C2C12, we find that inhibition of CAR3 (the mouse homolog of CA3) promotes CHRN internalization via a lipid raft-mediated pathway, leading to accelerated degradation of postsynaptic CHRN. Activation of CAR3 reduces CHRN degradation by suppressing receptor endocytosis. CAR3 exerts this effect by suppressing chaperone-assisted selective autophagy via interaction with BAG3 (BCL2-associated athanogene 3) and by dampening endoplasmic reticulum stress. Collectively, our study illustrates that skeletal muscle cell CAR3 is critical for CHRN homeostasis in the neuromuscular junction, and its deficiency leads to accelerated degradation of CHRN and development of myasthenia gravis, potentially revealing a novel therapeutic approach for this disorder.

  10. Investigation of optical properties and electronic transitions in bulk and nano-microribbons of molybdenum trioxide

    International Nuclear Information System (INIS)

    Lupan, O; Mishra, Y K; Adelung, R; Trofim, V; Cretu, V; Stamov, I; Syrbu, N N; Tiginyanu, I

    2014-01-01

    In this work, we report on crystalline quality and optical characteristics of molybdenum trioxide (MoO 3 ) bulk and nano-microribbons grown by rapid thermal oxidation (RTO). The developed RTO procedure allows one to synthesize highly crystalline (α-phase) bulk and nano-microribbons of MoO 3 . For R–Γ indirect transitions in bulk single crystals of MoO 3 , it has been found that the width of the bandgap along the E‖c polarization, associated with transitions R v1 –Γ c1 , is lower than the width of the band gap in polarization E ⊥ c, associated with transitions R v2 –Γ c2 . This result is indicative of splitting of the absorption edge due to α-MoO 3 structural anisotropy. Studies of the polarization dependence of the absorption in nano-microribbons (d ≈ 15–500 nm) demonstrated that the energy gap corresponding to R v1 –X c1 (E‖c) transition is smaller than that of R v2 –X c2 (E ⊥ c) transition. Similar dependence has been found for the R–Y indirect transitions. The results of the investigation of the reflectance spectra in the energy range from 3 to 6 eV are shown. By using the Kramers–Kronig method, the optical functions were derived from the reflection spectra of nano-microribbons, and the polarization dependence of direct energy transitions at the point R in the Brillouin zone are determined. The alternation in splitting caused by polarization of the absorption edge related to indirect transitions due to polarization opens new prospects for the design and fabricating interesting optoelectronic devices based on α-MoO 3 bulk and nano-microribbons with characteristics dependent on the polarization of light waves. (paper)

  11. Magnetizing of permanent magnet using HTS bulk magnet

    International Nuclear Information System (INIS)

    Oka, Tetsuo; Muraya, Tomoki; Kawasaki, Nobutaka; Fukui, Satoshi; Ogawa, Jun; Sato, Takao; Terasawa, Toshihisa

    2011-01-01

    A demagnetized Nd-Fe-B permanent magnet was scanned just above the magnetic pole containing the HTS bulk magnet, generating a magnetic field of 3.27 T. The magnet sample was subsequently found to be fully magnetized in the open space of the static magnetic fields. We examined the magnetic field distributions when the magnetic poles were scanned twice to activate the magnetic plates inversely with various overlap distances between the tracks of the HTS bulk magnet. The magnetic field of the 'rewritten' magnet reached the values of the magnetically saturated region of the material, showing steep gradients at the border of each magnetic pole. As a replacement for conventional pulse field magnetizing methods, this technique is proposed to expand the degree of freedom in the design of electromagnetic devices, and is proposed as a novel practical method for magnetizing rare-earth magnets, which have excellent magnetic performance and require intense fields of more than 3 T to be activated. (author)

  12. Investigation of the receptor-mediated endocytosis of transcobalamin/intrinsic factor-vitamin B12 complexes

    DEFF Research Database (Denmark)

    Beedholm, Rasmus; Grissom, Charles B.; Fedosov, Sergey N.

    receptor structure. This receptor is suggested to be regulated by the vitamin B12 level in the cells, which is interesting in relation to cancer growth. The cellular endocytosis of TC- B12 complex by this unknown receptor is being investigated, using confocal microscopy. Fluorescently labeled B12 molecules...... (Oregon green linked to B12) have been synthesized to determine the B12 uptake level in normal and various tumour-derived cells (e.g. Hela cells from cervix epithelioid carcinoma and BN- cells from rat yolk sac sarcoma). Costaining of the B12 binders has been performed using fluorescently labelled...

  13. Fluorescently labeled methyl-beta-cyclodextrin enters intestinal epithelial Caco-2 cells by fluid-phase endocytosis.

    Directory of Open Access Journals (Sweden)

    Ferenc Fenyvesi

    Full Text Available Cyclodextrins are widely used excipients for increasing the bioavailability of poorly water-soluble drugs. Their effect on drug absorption in the gastrointestinal tract is explained by their solubility- and permeability-enhancement. The aims of this study were to investigate penetration properties of fluorescently labeled randomly methylated-beta-cyclodextrin (FITC-RAMEB on Caco-2 cell layer and examine the cellular entry of cyclodextrins on intestinal cells. The permeability of FITC-RAMEB through Caco-2 monolayers was very limited. Using this compound in 0.05 mM concentration the permeability coefficient was 3.35±1.29×10(-8 cm/s and its permeability did not change in the presence of 5 mM randomly methylated-beta-cyclodextrin. Despite of the low permeability, cellular accumulation of FITC-RAMEB in cytoplasmic vesicles was significant and showed strong time and concentration dependence, similar to the characteristics of the macropinocytosis marker Lucifer Yellow. The internalization process was fully inhibited at 0°C and it was drastically reduced at 37°C applying rottlerin, an inhibitor of macropinocytosis. Notably, FITC-RAMEB colocalized with the early endosome organizer Rab5a. These results have revealed that FITC-RAMEB is able to enter intestinal epithelial cells by fluid-phase endocytosis from the apical side. This mechanism can be an additional process which helps to overcome the intestinal barrier and contributes to the bioavailability enhancement of cyclodextrins.

  14. Large area bulk superconductors

    Science.gov (United States)

    Miller, Dean J.; Field, Michael B.

    2002-01-01

    A bulk superconductor having a thickness of not less than about 100 microns is carried by a polycrystalline textured substrate having misorientation angles at the surface thereof not greater than about 15.degree.; the bulk superconductor may have a thickness of not less than about 100 microns and a surface area of not less than about 50 cm.sup.2. The textured substrate may have a thickness not less than about 10 microns and misorientation angles at the surface thereof not greater than about 15.degree.. Also disclosed is a process of manufacturing the bulk superconductor and the polycrystalline biaxially textured substrate material.

  15. Pou5f1-dependent EGF expression controls E-cad endocytosis, cell adhesion, and zebrafish epiboly movements

    Science.gov (United States)

    Song, Sungmin; Eckerle, Stephanie; Onichtchouk, Daria; Marrs, James A.; Nitschke, Roland; Driever, Wolfgang

    2013-01-01

    Summary Initiation of motile cell behavior in embryonic development occurs during late blastula stages when gastrulation begins. At this stage, the strong adhesion of blastomeres has to be modulated to enable dynamic behavior, similar to epithelial-to-mesenchymal transitions. We show that in zebrafish MZspg embryos mutant for the stem cell transcription factor Pou5f1/Oct4, which are severely delayed in the epiboly gastrulation movement, all blastomeres are defective in E-cad endosomal trafficking and E-cad accumulates at the plasma membrane. We find that Pou5f1-dependent control of EGF expression regulates endosomal E-cad trafficking. EGFR may act via modulation of p120 activity. Loss of E-cad dynamics reduces cohesion of cells in reaggregation assays. Quantitative analysis of cell behavior indicates that dynamic E-cad endosomal trafficking is required for epiboly cell movements. We hypothesize that dynamic control of E-cad trafficking is essential to effectively generate new adhesion sites when cells move relative to each other. PMID:23484854

  16. Synaptic activity regulates AMPA receptor trafficking through different recycling pathways

    Science.gov (United States)

    Zheng, Ning; Jeyifous, Okunola; Munro, Charlotte; Montgomery, Johanna M; Green, William N

    2015-01-01

    Changes in glutamatergic synaptic strength in brain are dependent on AMPA-type glutamate receptor (AMPAR) recycling, which is assumed to occur through a single local pathway. In this study, we present evidence that AMPAR recycling occurs through different pathways regulated by synaptic activity. Without synaptic stimulation, most AMPARs recycled in dynamin-independent endosomes containing the GTPase, Arf6. Few AMPARs recycled in dynamin-dependent endosomes labeled by transferrin receptors (TfRs). AMPAR recycling was blocked by alterations in the GTPase, TC10, which co-localized with Arf6 endosomes. TC10 mutants that reduced AMPAR recycling had no effect on increased AMPAR levels with long-term potentiation (LTP) and little effect on decreased AMPAR levels with long-term depression. However, internalized AMPAR levels in TfR-containing recycling endosomes increased after LTP, indicating increased AMPAR recycling through the dynamin-dependent pathway with synaptic plasticity. LTP-induced AMPAR endocytosis is inconsistent with local recycling as a source of increased surface receptors, suggesting AMPARs are trafficked from other sites. DOI: http://dx.doi.org/10.7554/eLife.06878.001 PMID:25970033

  17. Porphyrin-based Nanostructure-Dependent Photodynamic and Photothermal Therapies

    Science.gov (United States)

    Jin, Cheng S.

    This thesis presents the investigation of nanostructure-dependent phototherapy. We reviewed the liposomal structures for delivery of photosensitizers, and introduced a novel class of phototransducing liposomes called "porphysomes". Porphysomes are self-assembled from high packing density of pyropheophorbide alpha-conjugated phospholipids, resulting in extreme self-quenching of porphyrin fluorescence and comparable optical absorption to gold nanoparticles for high photothermal efficiency. We demonstrated this self-assembly of porphyrin-lipid conjugates converts a singlet oxygen generating mechanism (photodynamic therapy PDT activity) of porphyrin to photothermal mechanism (photothermal therapy PTT activity). The efficacy of porphysome-enhanced PTT was then evaluated on two pre-clinical animal models. We validated porphysome-enabled focal PTT to treat orthotopic prostate cancer using MRI-guided focal laser placement to closely mimic the current clinic procedure. Furthermore, porphysome-enabled fluorescence-guided transbronchial PTT of lung cancer was demonstrated in rabbit orthotopic lung cancer models, which led to the development of an ultra-minimally invasive therapy for early-stage peripheral lung cancer. On the other hand, the nanostructure-mediated conversion of PDT to PTT can be switched back by nanoparticle dissociation. By incorporating folate-conjugated phospholipids into the formulation, porphysomes were internalized into cells rapidly via folate receptor-mediated endocytosis and resulted in efficient disruption of nanostructures, which turned back on the photodynamic activity of densely packed porphyrins, making a closed loop of conversion between PDT and PTT. The multimodal imaging and therapeutic features of porphysome make it ideal for future personalized cancer treatments.

  18. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Ji Hye; Joo, Sang-Woo [Department of Chemistry, Soongsil University, Seoul 156-743 (Korea, Republic of); Cho, Keunchang [Logos Biosystems, Incorporated, Anyang 431-070 (Korea, Republic of); Lee, So Yeong, E-mail: leeso@snu.ac.kr, E-mail: sjoo@ssu.ac.kr [Laboratory of Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742 (Korea, Republic of)

    2011-06-10

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  19. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    International Nuclear Information System (INIS)

    Seo, Ji Hye; Joo, Sang-Woo; Cho, Keunchang; Lee, So Yeong

    2011-01-01

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  20. Singularities and Entropy in Bulk Viscosity Dark Energy Model

    International Nuclear Information System (INIS)

    Meng Xinhe; Dou Xu

    2011-01-01

    In this paper bulk viscosity is introduced to describe the effects of cosmic non-perfect fluid on the cosmos evolution and to build the unified dark energy (DE) with (dark) matter models. Also we derive a general relation between the bulk viscosity form and Hubble parameter that can provide a procedure for the viscosity DE model building. Especially, a redshift dependent viscosity parameter ζ ∝ λ 0 + λ 1 (1 + z) n proposed in the previous work [X.H. Meng and X. Dou, Commun. Theor. Phys. 52 (2009) 377] is investigated extensively in this present work. Further more we use the recently released supernova dataset (the Constitution dataset) to constrain the model parameters. In order to differentiate the proposed concrete dark energy models from the well known ΛCDM model, statefinder diagnostic method is applied to this bulk viscosity model, as a complementary to the Om parameter diagnostic and the deceleration parameter analysis performed by us before. The DE model evolution behavior and tendency are shown in the plane of the statefinder diagnostic parameter pair {r, s} as axes where the fixed point represents the ΛCDM model. The possible singularity property in this bulk viscosity cosmology is also discussed to which we can conclude that in the different parameter regions chosen properly, this concrete viscosity DE model can have various late evolution behaviors and the late time singularity could be avoided. We also calculate the cosmic entropy in the bulk viscosity dark energy frame, and find that the total entropy in the viscosity DE model increases monotonously with respect to the scale factor evolution, thus this monotonous increasing property can indicate an arrow of time in the universe evolution, though the quantum version of the arrow of time is still very puzzling. (geophysics, astronomy, and astrophysics)

  1. Thermodynamic Derivation of the Activation Energy for Ice Nucleation

    Science.gov (United States)

    Barahona, D.

    2015-01-01

    Cirrus clouds play a key role in the radiative and hydrological balance of the upper troposphere. Their correct representation in atmospheric models requires an understanding of the microscopic processes leading to ice nucleation. A key parameter in the theoretical description of ice nucleation is the activation energy, which controls the flux of water molecules from the bulk of the liquid to the solid during the early stages of ice formation. In most studies it is estimated by direct association with the bulk properties of water, typically viscosity and self-diffusivity. As the environment in the ice-liquid interface may differ from that of the bulk, this approach may introduce bias in calculated nucleation rates. In this work a theoretical model is proposed to describe the transfer of water molecules across the ice-liquid interface. Within this framework the activation energy naturally emerges from the combination of the energy required to break hydrogen bonds in the liquid, i.e., the bulk diffusion process, and the work dissipated from the molecular rearrangement of water molecules within the ice-liquid interface. The new expression is introduced into a generalized form of classical nucleation theory. Even though no nucleation rate measurements are used to fit any of the parameters of the theory the predicted nucleation rate is in good agreement with experimental results, even at temperature as low as 190 K, where it tends to be underestimated by most models. It is shown that the activation energy has a strong dependency on temperature and a weak dependency on water activity. Such dependencies are masked by thermodynamic effects at temperatures typical of homogeneous freezing of cloud droplets; however, they may affect the formation of ice in haze aerosol particles. The new model provides an independent estimation of the activation energy and the homogeneous ice nucleation rate, and it may help to improve the interpretation of experimental results and the

  2. Superoxide dismutase and catalase conjugated to polyethylene glycol increases endothelial enzyme activity and oxidant resistance

    International Nuclear Information System (INIS)

    Beckman, J.S.; Minor, R.L. Jr.; White, C.W.; Repine, J.E.; Rosen, G.M.; Freeman, B.A.

    1988-01-01

    Covalent conjugation of superoxide dismutase and catalase with polyethylene glycol (PEG) increases the circulatory half-lives of these enzymes from 125 I-PEG-catalase or 125 I-PEG-superoxide dismutase produced a linear, concentration-dependent increase in cellular enzyme activity and radioactivity. Fluorescently labeled PEG-superoxide dismutase incubated with endothelial cells showed a vesicular localization. Mechanical injury to cell monolayers, which is known to stimulate endocytosis, further increased the uptake of fluorescent PEG-superoxide dismutase. Addition of PEG and PEG-conjugated enzymes perturbed the spin-label binding environment, indicative of producing an increase in plasma membrane fluidity. Thus, PEG conjugation to superoxide dismutase and catalase enhances cell association of these enzymes in a manner which increases cellular enzyme activities and provides prolonged protection from partially reduced oxygen species

  3. Bioaccumulation and oxidative stress responses measured in the estuarine ragworm (Nereis diversicolor) exposed to dissolved, nano- and bulk-sized silver

    International Nuclear Information System (INIS)

    Cozzari, Margherita; Elia, Antonia Concetta; Pacini, Nicole; Smith, Brian D.; Boyle, David; Rainbow, Philip S.; Khan, Farhan R.

    2015-01-01

    The impact of Ag NPs on sediment-dwelling organisms has received relatively little attention, particularly in linking bioaccumulation to oxidative injury. The polychaete Nereis diversicolor was exposed to sediments spiked with dissolved Ag (added as AgNO 3 ), Ag NPs (63 ± 27 nm) and larger bulk Ag particles (202 ± 56 μm), for up to 11 days at sublethal concentrations (nominally 2.5, 5, 10 μg Ag g −1 sediment (dw)). There were concentration- and time-dependent differences in the accumulation of the three Ag forms, but all three forms elicited an oxidative stress response. In the cases of Ag NPs and bulk Ag particles, changes in antioxidant markers (glutathione, SOD, CAT, GPx, SeGPx, GST and GR) occurred without significant Ag accumulation. Differences in biomarker profiles between the three Ag forms suggest that the mechanism of oxidative stress caused by particulate Ag is distinct from that of dissolved Ag. - Highlights: • N. diversicolor exposed to dissolved, nano and bulk Ag via spiked sediments. • Concentration- and time-dependent differences in the accumulation patterns. • All three forms elicited an oxidative stress response. • Antioxidant markers changed without significant accumulation for particulate Ag. • Biomarker profile resulting from particulate Ag exposure different to dissolved Ag. - Exposure to different Ag forms leads to distinct oxidative stress biomarker profiles. Particulate Ag activates the oxidative stress response differently to dissolved Ag

  4. Superconductivity in Bi-Sr-Ca-Cu-O bulk and film ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Sukhanov, A A; Ozmanian, KH R; Sandomirskii, B B

    1988-07-01

    A superconducting transition with Tc0 = 82-95 K and Tc(R = 0) = 82-72 K was observed in Bi(Sr/1-x/Ca/x/)2Cu3O(y) bulk and film specimens obtained via a solid-phase reaction. Temperature dependences of the resistance, critical current, and magnetic susceptibility were measured.

  5. Linear magnetoresistance and surface to bulk coupling in topological insulator thin films.

    Science.gov (United States)

    Singh, Sourabh; Gopal, R K; Sarkar, Jit; Pandey, Atul; Patel, Bhavesh G; Mitra, Chiranjib

    2017-12-20

    We explore the temperature dependent magnetoresistance of bulk insulating topological insulator thin films. Thin films of Bi 2 Se 2 Te and BiSbTeSe 1.6 were grown using the pulsed laser deposition technique and subjected to transport measurements. Magnetotransport measurements indicate a non-saturating linear magnetoresistance (LMR) behavior at high magnetic field values. We present a careful analysis to explain the origin of LMR taking into consideration all the existing models of LMR. Here we consider that the bulk insulating states and the metallic surface states constitute two parallel conduction channels. Invoking this, we were able to explain linear magnetoresistance behavior as a competition between these parallel channels. We observe that the cross-over field, where LMR sets in, decreases with increasing temperature. We propose that this cross-over field can be used phenomenologically to estimate the strength of surface to bulk coupling.

  6. Anisotropic Bianchi Type-III Bulk Viscous Fluid Universe in Lyra Geometry

    Directory of Open Access Journals (Sweden)

    Priyanka Kumari

    2013-01-01

    Full Text Available An anisotropic Bianchi type-III cosmological model is investigated in the presence of a bulk viscous fluid within the framework of Lyra geometry with time-dependent displacement vector. It is shown that the field equations are solvable for any arbitrary function of a scale factor. To get the deterministic model of the universe, we have assumed that (i a simple power-law form of a scale factor and (ii the bulk viscosity coefficient are proportional to the energy density of the matter. The exact solutions of the Einstein’s field equations are obtained which represent an expanding, shearing, and decelerating model of the universe. Some physical and kinematical behaviors of the cosmological model are briefly discussed.

  7. Activity-Dependent Bidirectional Regulation of GAD Expression in a Homeostatic Fashion Is Mediated by BDNF-Dependent and Independent Pathways

    Science.gov (United States)

    Hanno-Iijima, Yoko; Tanaka, Masami; Iijima, Takatoshi

    2015-01-01

    Homeostatic synaptic plasticity, or synaptic scaling, is a mechanism that tunes neuronal transmission to compensate for prolonged, excessive changes in neuronal activity. Both excitatory and inhibitory neurons undergo homeostatic changes based on synaptic transmission strength, which could effectively contribute to a fine-tuning of circuit activity. However, gene regulation that underlies homeostatic synaptic plasticity in GABAergic (GABA, gamma aminobutyric) neurons is still poorly understood. The present study demonstrated activity-dependent dynamic scaling in which NMDA-R (N-methyl-D-aspartic acid receptor) activity regulated the expression of GABA synthetic enzymes: glutamic acid decarboxylase 65 and 67 (GAD65 and GAD67). Results revealed that activity-regulated BDNF (brain-derived neurotrophic factor) release is necessary, but not sufficient, for activity-dependent up-scaling of these GAD isoforms. Bidirectional forms of activity-dependent GAD expression require both BDNF-dependent and BDNF-independent pathways, both triggered by NMDA-R activity. Additional results indicated that these two GAD genes differ in their responsiveness to chronic changes in neuronal activity, which could be partially caused by differential dependence on BDNF. In parallel to activity-dependent bidirectional scaling in GAD expression, the present study further observed that a chronic change in neuronal activity leads to an alteration in neurotransmitter release from GABAergic neurons in a homeostatic, bidirectional fashion. Therefore, the differential expression of GAD65 and 67 during prolonged changes in neuronal activity may be implicated in some aspects of bidirectional homeostatic plasticity within mature GABAergic presynapses. PMID:26241953

  8. Quasiparticle interference of the Fermi arcs and surface-bulk connectivity of a Weyl semimetal.

    Science.gov (United States)

    Inoue, Hiroyuki; Gyenis, András; Wang, Zhijun; Li, Jian; Oh, Seong Woo; Jiang, Shan; Ni, Ni; Bernevig, B Andrei; Yazdani, Ali

    2016-03-11

    Weyl semimetals host topologically protected surface states, with arced Fermi surface contours that are predicted to propagate through the bulk when their momentum matches that of the surface projections of the bulk's Weyl nodes. We used spectroscopic mapping with a scanning tunneling microscope to visualize quasiparticle scattering and interference at the surface of the Weyl semimetal TaAs. Our measurements reveal 10 different scattering wave vectors, which can be understood and precisely reproduced with a theory that takes into account the shape, spin texture, and momentum-dependent propagation of the Fermi arc surface states into the bulk. Our findings provide evidence that Weyl nodes act as sinks for electron transport on the surface of these materials. Copyright © 2016, American Association for the Advancement of Science.

  9. Application of textured YBCO bulks with artificial holes for superconducting magnetic bearing

    International Nuclear Information System (INIS)

    Dias, D H N; Sotelo, G G; Moysés, L A; Telles, L G T; Bernstein, P; Aburas, M; Noudem, J G; Kenfaui, D; Chaud, X

    2015-01-01

    The levitation force between a superconductor and a permanent magnet has been investigated for the development of superconducting magnetic bearings (SMBs). Depending on the proposed application, the SMBs can be arranged with two kinds of symmetries: rotational or linear. The SMBs present passive operation, low level of noise and no friction, but they need a cooling system for their operation. Nowadays the cooling problem may be easily solved by the use of a commercial cryocooler. The levitation force of SMBs is directly related to the quality of the superconductor material (which depends on its critical current density) and the permanent magnet arrangement. Also, research about the YBa 2 Cu 3 O x (Y123) bulk materials has shown that artificial holes enhance the superconducting properties, in particular the magnetic trapped field. In this context, this work proposes the investigation of the levitation force of a bulk Y123 sample with multiple holes and the comparison of its performances with those of conventional plain Y123 superconductors. (paper)

  10. BACE1 protein endocytosis and trafficking are differentially regulated by ubiquitination at lysine 501 and the Di-leucine motif in the carboxyl terminus.

    Science.gov (United States)

    Kang, Eugene L; Biscaro, Barbara; Piazza, Fabrizio; Tesco, Giuseppina

    2012-12-14

    β-Site amyloid precursor protein-cleaving enzyme (BACE1) is a membrane-tethered member of the aspartyl proteases that has been identified as β-secretase. BACE1 is targeted through the secretory pathway to the plasma membrane and then is internalized to endosomes. Sorting of membrane proteins to the endosomes and lysosomes is regulated by the interaction of signals present in their carboxyl-terminal fragment with specific trafficking molecules. The BACE1 carboxyl-terminal fragment contains a di-leucine sorting signal ((495)DDISLL(500)) and a ubiquitination site at Lys-501. Here, we report that lack of ubiquitination at Lys-501 (BACE1K501R) does not affect the rate of endocytosis but produces BACE1 stabilization and accumulation of BACE1 in early and late endosomes/lysosomes as well as at the cell membrane. In contrast, the disruption of the di-leucine motif (BACE1LLAA) greatly impairs BACE1 endocytosis and produces a delayed retrograde transport of BACE1 to the trans-Golgi network (TGN) and a delayed delivery of BACE1 to the lysosomes, thus decreasing its degradation. Moreover, the combination of the lack of ubiquitination at Lys-501 and the disruption of the di-leucine motif (BACE1LLAA/KR) produces additive effects on BACE1 stabilization and defective internalization. Finally, BACE1LLAA/KR accumulates in the TGN, while its levels are decreased in EEA1-positive compartments indicating that both ubiquitination at Lys-501 and the di-leucine motif are necessary for the trafficking of BACE1 from the TGN to early endosomes. Our studies have elucidated a differential role for the di-leucine motif and ubiquitination at Lys-501 in BACE1 endocytosis, trafficking, and degradation and suggest the involvement of multiple adaptor molecules.

  11. BACE1 Protein Endocytosis and Trafficking Are Differentially Regulated by Ubiquitination at Lysine 501 and the Di-leucine Motif in the Carboxyl Terminus*

    Science.gov (United States)

    Kang, Eugene L.; Biscaro, Barbara; Piazza, Fabrizio; Tesco, Giuseppina

    2012-01-01

    β-Site amyloid precursor protein-cleaving enzyme (BACE1) is a membrane-tethered member of the aspartyl proteases that has been identified as β-secretase. BACE1 is targeted through the secretory pathway to the plasma membrane and then is internalized to endosomes. Sorting of membrane proteins to the endosomes and lysosomes is regulated by the interaction of signals present in their carboxyl-terminal fragment with specific trafficking molecules. The BACE1 carboxyl-terminal fragment contains a di-leucine sorting signal (495DDISLL500) and a ubiquitination site at Lys-501. Here, we report that lack of ubiquitination at Lys-501 (BACE1K501R) does not affect the rate of endocytosis but produces BACE1 stabilization and accumulation of BACE1 in early and late endosomes/lysosomes as well as at the cell membrane. In contrast, the disruption of the di-leucine motif (BACE1LLAA) greatly impairs BACE1 endocytosis and produces a delayed retrograde transport of BACE1 to the trans-Golgi network (TGN) and a delayed delivery of BACE1 to the lysosomes, thus decreasing its degradation. Moreover, the combination of the lack of ubiquitination at Lys-501 and the disruption of the di-leucine motif (BACE1LLAA/KR) produces additive effects on BACE1 stabilization and defective internalization. Finally, BACE1LLAA/KR accumulates in the TGN, while its levels are decreased in EEA1-positive compartments indicating that both ubiquitination at Lys-501 and the di-leucine motif are necessary for the trafficking of BACE1 from the TGN to early endosomes. Our studies have elucidated a differential role for the di-leucine motif and ubiquitination at Lys-501 in BACE1 endocytosis, trafficking, and degradation and suggest the involvement of multiple adaptor molecules. PMID:23109336

  12. AS160 associates with the Na+,K+-ATPase and mediates the adenosine monophosphate-stimulated protein kinase-dependent regulation of sodium pump surface expression.

    Science.gov (United States)

    Alves, Daiane S; Farr, Glen A; Seo-Mayer, Patricia; Caplan, Michael J

    2010-12-01

    The Na(+),K(+)-ATPase is the major active transport protein found in the plasma membranes of most epithelial cell types. The regulation of Na(+),K(+)-ATPase activity involves a variety of mechanisms, including regulated endocytosis and recycling. Our efforts to identify novel Na(+),K(+)-ATPase binding partners revealed a direct association between the Na(+),K(+)-ATPase and AS160, a Rab-GTPase-activating protein. In COS cells, coexpression of AS160 and Na(+),K(+)-ATPase led to the intracellular retention of the sodium pump. We find that AS160 interacts with the large cytoplasmic NP domain of the α-subunit of the Na(+),K(+)-ATPase. Inhibition of the activity of the adenosine monophosphate-stimulated protein kinase (AMPK) in Madin-Darby canine kidney cells through treatment with Compound C induces Na(+),K(+)-ATPase endocytosis. This effect of Compound C is prevented through the short hairpin RNA-mediated knockdown of AS160, demonstrating that AMPK and AS160 participate in a common pathway to modulate the cell surface expression of the Na(+),K(+)-ATPase.

  13. A CCR2 macrophage endocytic pathway mediates extravascular fibrin clearance in vivo

    DEFF Research Database (Denmark)

    Motley, Michael P; Madsen, Daniel H; Jürgensen, Henrik J

    2016-01-01

    cellular endocytosis and lysosomal targeting, revealing a novel intracellular pathway for extravascular fibrin degradation. A C-C chemokine receptor type 2 (CCR2)-positive macrophage subpopulation constituted the majority of fibrin-uptaking cells. Consequently, cellular fibrin uptake was diminished...... by elimination of CCR2-expressing cells. The CCR2-positive macrophage subtype was different from collagen-internalizing M2-like macrophages. Cellular fibrin uptake was strictly dependent on plasminogen and plasminogen activator. Surprisingly, however, fibrin endocytosis was unimpeded by the absence of the fibrin...... subsets of macrophages employing distinct molecular pathways....

  14. Developing bulk exchange spring magnets

    Science.gov (United States)

    Mccall, Scott K.; Kuntz, Joshua D.

    2017-06-27

    A method of making a bulk exchange spring magnet by providing a magnetically soft material, providing a hard magnetic material, and producing a composite of said magnetically soft material and said hard magnetic material to make the bulk exchange spring magnet. The step of producing a composite of magnetically soft material and hard magnetic material is accomplished by electrophoretic deposition of the magnetically soft material and the hard magnetic material to make the bulk exchange spring magnet.

  15. Pressure dependence of the elastic constants and vibrational anharmonicity of Pd sub 3 sub 9 Ni sub 1 sub 0 Cu sub 3 sub 0 P sub 2 sub 1 bulk metallic glass

    CERN Document Server

    Wang Li; Sun, L L; Wang, W H; Wang, W K

    2003-01-01

    The pressure dependence of the acoustic velocities of a Pd sub 3 sub 9 Ni sub 1 sub 0 Cu sub 3 sub 0 P sub 2 sub 1 bulk metallic glass have been investigated up to 0.5 GPa at room temperature with the pulse echo overlap method. Two independent second-order elastic coefficients C sub 1 sub 1 and C sub 4 sub 4 and their pressure derivatives are yielded. The vibrational anharmonicity is shown by calculating both the acoustic mode Grueneisen parameters in the long-wavelength limit and the thermal Grueneisen parameter, and this result is compared with that for the Pd sub 4 sub 0 Ni sub 4 sub 0 P sub 2 sub 0 bulk glass.

  16. Transient regimes during high-temperature deformation of a bulk metallic glass: A free volume approach

    International Nuclear Information System (INIS)

    Bletry, M.; Guyot, P.; Brechet, Y.; Blandin, J.J.; Soubeyroux, J.L.

    2007-01-01

    The homogeneous deformation of a zirconium-based bulk metallic glass is investigated in the glass transition range. Compression and stress-relaxation tests have been conducted. The stress-strain curves are modeled in the framework of the free volume theory, including transient phenomena (overshoot and undershoot). This approach allows several physical parameters (activation volume, flow defect creation and relaxation coefficient) to be determined from a mechanical experiment. This model is able to rationalize the dependency of stress overshoot on relaxation time. It is shown that, due to the relationship between flow defect concentration and free volume model, it is impossible to determine the equilibrium flow defect concentration. However, the relative variation of flow defect is always the same, and all the model parameters depend on the equilibrium flow defect concentration. The methodology presented in this paper should, in the future, allow the consistency of the free volume model to be assessed

  17. Morphological changes of plasma membrane and protein assembly during clathrin-mediated endocytosis

    Science.gov (United States)

    Yoshida, Aiko; Sakai, Nobuaki; Uekusa, Yoshitsugu; Imaoka, Yuka; Itagaki, Yoshitsuna; Suzuki, Yuki

    2018-01-01

    Clathrin-mediated endocytosis (CME) proceeds through a series of morphological changes of the plasma membrane induced by a number of protein components. Although the spatiotemporal assembly of these proteins has been elucidated by fluorescence-based techniques, the protein-induced morphological changes of the plasma membrane have not been fully clarified in living cells. Here, we visualize membrane morphology together with protein localizations during CME by utilizing high-speed atomic force microscopy (HS-AFM) combined with a confocal laser scanning unit. The plasma membrane starts to invaginate approximately 30 s after clathrin starts to assemble, and the aperture diameter increases as clathrin accumulates. Actin rapidly accumulates around the pit and induces a small membrane swelling, which, within 30 s, rapidly covers the pit irreversibly. Inhibition of actin turnover abolishes the swelling and induces a reversible open–close motion of the pit, indicating that actin dynamics are necessary for efficient and irreversible pit closure at the end of CME. PMID:29723197

  18. How synthetic membrane systems contribute to the understanding of lipid-driven endocytosis.

    Science.gov (United States)

    Schubert, Thomas; Römer, Winfried

    2015-11-01

    Synthetic membrane systems, such as giant unilamellar vesicles and solid supported lipid bilayers, have widened our understanding of biological processes occurring at or through membranes. Artificial systems are particularly suited to study the inherent properties of membranes with regard to their components and characteristics. This review critically reflects the emerging molecular mechanism of lipid-driven endocytosis and the impact of model membrane systems in elucidating the complex interplay of biomolecules within this process. Lipid receptor clustering induced by binding of several toxins, viruses and bacteria to the plasma membrane leads to local membrane bending and formation of tubular membrane invaginations. Here, lipid shape, and protein structure and valency are the essential parameters in membrane deformation. Combining observations of complex cellular processes and their reconstitution on minimal systems seems to be a promising future approach to resolve basic underlying mechanisms. This article is part of a Special Issue entitled: Mechanobiology. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Copper phthalocyanine and metal free phthalocyanine bulk heterojunction photodetector

    Energy Technology Data Exchange (ETDEWEB)

    Farooq, Amjad, E-mail: amjad.farooq1212@hotmail.com [Wah Engineering College, University of Wah, Wah Cantt. 47040 (Pakistan); GIK Institute of Engineering Sciences and Technology, Topi 23640, Swabi (Pakistan); Karimov, Kh.S. [GIK Institute of Engineering Sciences and Technology, Topi 23640, Swabi (Pakistan); Physical Technical Institute, Aini St. 299/1, Dushanbe 734063 (Tajikistan); Ahmed, Nisar; Ali, Taimoor [GIK Institute of Engineering Sciences and Technology, Topi 23640, Swabi (Pakistan); Khalid Alamgir, M. [National Institute of Vacuum Science and Technology, NCP complex, Islamabad 44000 (Pakistan); Usman, Muhammad [Experimental Physics Laboratories, National Centre for Physics, Quaid-i-Azam University, Islamabad 44000 (Pakistan)

    2015-01-15

    In this study we present the dependence of electrical properties of copper phthalocyanine (CuPc) and metal free phthalocyanine (H{sub 2}Pc) bulk heterojunction structure under different illumination levels. To fabricate the device on ITO coated glass substrate the bulk heterojunction thin film of CuPc and H{sub 2}Pc with thickness varying from 100 nm to 300 nm are deposited by thermal evaporator. Aluminum thin film was deposited by thermal evaporation as a top contact. The optical properties of the fabricated device are investigated using UV–vis spectroscopy. The current-voltage characteristics in dark and under illumination show that the device is sensitive towards visible light. The absorption spectrum describes its photo sensitivity in the range of wavelength from 200 nm to 850 nm. Simulation of current-intensity of light curve is carried out and experimental results are found in good agreement with simulated ones.

  20. Copper phthalocyanine and metal free phthalocyanine bulk heterojunction photodetector

    International Nuclear Information System (INIS)

    Farooq, Amjad; Karimov, Kh.S.; Ahmed, Nisar; Ali, Taimoor; Khalid Alamgir, M.; Usman, Muhammad

    2015-01-01

    In this study we present the dependence of electrical properties of copper phthalocyanine (CuPc) and metal free phthalocyanine (H 2 Pc) bulk heterojunction structure under different illumination levels. To fabricate the device on ITO coated glass substrate the bulk heterojunction thin film of CuPc and H 2 Pc with thickness varying from 100 nm to 300 nm are deposited by thermal evaporator. Aluminum thin film was deposited by thermal evaporation as a top contact. The optical properties of the fabricated device are investigated using UV–vis spectroscopy. The current-voltage characteristics in dark and under illumination show that the device is sensitive towards visible light. The absorption spectrum describes its photo sensitivity in the range of wavelength from 200 nm to 850 nm. Simulation of current-intensity of light curve is carried out and experimental results are found in good agreement with simulated ones

  1. Significance of settling model structures and parameter subsets in modelling WWTPs under wet-weather flow and filamentous bulking conditions

    DEFF Research Database (Denmark)

    Ramin, Elham; Sin, Gürkan; Mikkelsen, Peter Steen

    2014-01-01

    Current research focuses on predicting and mitigating the impacts of high hydraulic loadings on centralized wastewater treatment plants (WWTPs) under wet-weather conditions. The maximum permissible inflow to WWTPs depends not only on the settleability of activated sludge in secondary settling tanks...... (SSTs) but also on the hydraulic behaviour of SSTs. The present study investigates the impacts of ideal and non-ideal flow (dry and wet weather) and settling (good settling and bulking) boundary conditions on the sensitivity of WWTP model outputs to uncertainties intrinsic to the one-dimensional (1-D...... of settling parameters to the total variance of the key WWTP process outputs significantly depends on the influent flow and settling conditions. The magnitude of the impact is found to vary, depending on which type of 1-D SST model is used. Therefore, we identify and recommend potential parameter subsets...

  2. Modelling of bulk superconductor magnetization

    International Nuclear Information System (INIS)

    Ainslie, M D; Fujishiro, H

    2015-01-01

    This paper presents a topical review of the current state of the art in modelling the magnetization of bulk superconductors, including both (RE)BCO (where RE = rare earth or Y) and MgB 2 materials. Such modelling is a powerful tool to understand the physical mechanisms of their magnetization, to assist in interpretation of experimental results, and to predict the performance of practical bulk superconductor-based devices, which is particularly important as many superconducting applications head towards the commercialization stage of their development in the coming years. In addition to the analytical and numerical techniques currently used by researchers for modelling such materials, the commonly used practical techniques to magnetize bulk superconductors are summarized with a particular focus on pulsed field magnetization (PFM), which is promising as a compact, mobile and relatively inexpensive magnetizing technique. A number of numerical models developed to analyse the issues related to PFM and optimise the technique are described in detail, including understanding the dynamics of the magnetic flux penetration and the influence of material inhomogeneities, thermal properties, pulse duration, magnitude and shape, and the shape of the magnetization coil(s). The effect of externally applied magnetic fields in different configurations on the attenuation of the trapped field is also discussed. A number of novel and hybrid bulk superconductor structures are described, including improved thermal conductivity structures and ferromagnet–superconductor structures, which have been designed to overcome some of the issues related to bulk superconductors and their magnetization and enhance the intrinsic properties of bulk superconductors acting as trapped field magnets. Finally, the use of hollow bulk cylinders/tubes for shielding is analysed. (topical review)

  3. Mining the bulk positron lifetime

    International Nuclear Information System (INIS)

    Aourag, H.; Guittom, A.

    2009-01-01

    We introduce a new approach to investigate the bulk positron lifetimes of new systems based on data-mining techniques. Through data mining of bulk positron lifetimes, we demonstrate the ability to predict the positron lifetimes of new semiconductors on the basis of available semiconductor data already studied. Informatics techniques have been applied to bulk positron lifetimes for different tetrahedrally bounded semiconductors in order to discover computational design rules. (copyright 2009 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  4. Activity-dependent branching ratios in stocks, solar x-ray flux, and the Bak-Tang-Wiesenfeld sandpile model

    Science.gov (United States)

    Martin, Elliot; Shreim, Amer; Paczuski, Maya

    2010-01-01

    We define an activity-dependent branching ratio that allows comparison of different time series Xt . The branching ratio bx is defined as bx=E[ξx/x] . The random variable ξx is the value of the next signal given that the previous one is equal to x , so ξx={Xt+1∣Xt=x} . If bx>1 , the process is on average supercritical when the signal is equal to x , while if bxmarket hypothesis.” For stock volumes, solar x-ray flux intensities, and the Bak-Tang-Wiesenfeld (BTW) sandpile model, bx is supercritical for small values of activity and subcritical for the largest ones, indicating a tendency to return to a typical value. For stock volumes this tendency has an approximate power-law behavior. For solar x-ray flux and the BTW model, there is a broad regime of activity where bx≃1 , which we interpret as an indicator of critical behavior. This is true despite different underlying probability distributions for Xt and for ξx . For the BTW model the distribution of ξx is Gaussian, for x sufficiently larger than 1, and its variance grows linearly with x . Hence, the activity in the BTW model obeys a central limit theorem when sampling over past histories. The broad region of activity where bx is close to one disappears once bulk dissipation is introduced in the BTW model—supporting our hypothesis that it is an indicator of criticality.

  5. Molecular Intercalation and Cohesion of Organic Bulk Heterojunction Photovoltaic Devices

    KAUST Repository

    Bruner, Christopher; Miller, Nichole C.; McGehee, Michael D.; Dauskardt, Reinhold H.

    2013-01-01

    The phase separated bulk heterojunction (BHJ) layer in BHJ polymer:fullerene organic photovoltaic devices (OPV) are mechanically weak with low values of cohesion. Improved cohesion is important for OPV device thermomechanical reliability. BHJ devices are investigated and how fullerene intercalation within the active layer affects cohesive properties in the BHJ is shown. The intercalation of fullerenes between the side chains of the polymers poly(3,3″′-didocecyl quaterthiophene) (PQT-12) and poly(2,5-bis(3-hexadecylthiophen-2-yl)thieno[3,2-b]thiophene (pBTTT) is shown to enhance BHJ layer cohesion. Cohesion values range from ≈1 to 5 J m -2, depending on the polymer:fullerene blend, processing conditions, and composition. Devices with non-intercalated BHJ layers are found to have significantly reduced values of cohesion. The resulting device power conversion efficiencies (PCE) are also investigated and correlated with the device cohesion. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Molecular Intercalation and Cohesion of Organic Bulk Heterojunction Photovoltaic Devices

    KAUST Repository

    Bruner, Christopher

    2013-01-17

    The phase separated bulk heterojunction (BHJ) layer in BHJ polymer:fullerene organic photovoltaic devices (OPV) are mechanically weak with low values of cohesion. Improved cohesion is important for OPV device thermomechanical reliability. BHJ devices are investigated and how fullerene intercalation within the active layer affects cohesive properties in the BHJ is shown. The intercalation of fullerenes between the side chains of the polymers poly(3,3″′-didocecyl quaterthiophene) (PQT-12) and poly(2,5-bis(3-hexadecylthiophen-2-yl)thieno[3,2-b]thiophene (pBTTT) is shown to enhance BHJ layer cohesion. Cohesion values range from ≈1 to 5 J m -2, depending on the polymer:fullerene blend, processing conditions, and composition. Devices with non-intercalated BHJ layers are found to have significantly reduced values of cohesion. The resulting device power conversion efficiencies (PCE) are also investigated and correlated with the device cohesion. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Effects of gypsum and bulk density on neutron probe calibration curves

    International Nuclear Information System (INIS)

    Arslan, Awadis; Razzouk, A.K.

    1993-10-01

    The effects of gypsum and bulk density on the neutron probe calibration curve were studied in the laboratory and in the field. The effect of bulk density was negligible for the soil studied in the laboratory, while it was significant for the field calibration. An increase in the slope of moisture content on a volume basis vs. count ratio with increasing gypsum content at the soil was observed in the laboratory calibration. A simple method for correction of the calibration curve for gypsum content was adopted to obtain a specific curve for each layer. The adapted method requires the gypsum fraction to be estimated for each layer and then incorporated in the calibration curve to improve the coefficient of determination. A field calibration showed an improvement of the determination coefficient by introducing bulk density and gypsum fraction, in addition to count ratio using moisture content on a volume basis as a dependent variable in multi linear regression analysis. The same procedure was successful with variable gravel fractions. (author). 18 refs., 3 figs., 2 tabs

  8. Potential benefits and phytotoxicity of bulk and nano-chitosan on the growth, morphogenesis, physiology, and micropropagation of Capsicum annuum.

    Science.gov (United States)

    Asgari-Targhi, Ghasem; Iranbakhsh, Alireza; Ardebili, Zahra Oraghi

    2018-06-01

    Concerning environmental issues of metal based-nanomaterials and increasing demand for nano-based products; various strategies have been employed to find eco-friendly natural nano-compounds, among which nano-polymer chitosan is mostly considered. Herein, the various aspects of the way in which bulk or nano-chitosan may modify growth, morphogenesis, micropropagation, and physiology of Capsicum annuum L. were considered. Culture medium was manipulated with different concentrations of bulk chitosan or synthesized chitosan/tripolyphosphate (TPP) nano-particle. The supplementations of culture media led to changes in morphology (especially, the root architecture) and differentiation. Toxic doses of bulk (100 mgL -1 ) or nano-chitosan (5, 10, and 20 mgL -1 ) dramatically provoked cessation of plant growth and development. Plant growth and biomass accumulations were increased along with the suitable levels of bulk or nano-chitosan. Peroxidase and catalase activities in a dose and organ-dependent manners were significantly modified by the supplements. Phenylalanine ammonia lyase was induced by the mentioned supplements. Also, the contents of soluble phenols, proline, and alkaloid were found to be significantly increased by the elicitors, over the control. The nano-chitosan of 1 mgL -1 was found to be the most effective elicitor to trigger organogenesis via micropropagation. The huge differences between triggering and toxic concentrations of the supplements would be due to the physicochemical modifications of nano-polymeric. Furthermore, the results highlight the potential benefits (hormone-like activity) and phytotoxic impacts of nano-chitosan/TPP for in vitro manipulations. This is the first report on both the favorable and adverse effects of nano-chitosan/TPP, representing requirements for further investigation on such formulations for future applications. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  9. Nicotine dependence, physical activity, and sedentary behavior among adult smokers

    OpenAIRE

    Paul D Loprinzi; Jerome F Walker

    2015-01-01

    Background: Research has previously demonstrated an inverse association between smoking status and physical activity; however, few studies have examined the association between nicotine dependence and physical activity or sedentary behavior. Aim: This study examined the association between nicotine dependence and accelerometer-determined physical activity and sedentary behavior. Materials and Methods: Data from the 2003-2006 National Health and Nutrition Examination Survey (NHANES) were used....

  10. Dependence of Exciton Diffusion Length and Diffusion Coefficient on Photophysical Parameters in Bulk Heterojunction Organic Solar Cells

    Science.gov (United States)

    Yeboah, Douglas; Singh, Jai

    2017-11-01

    Recently, the dependence of exciton diffusion length (LD ) on some photophysical parameters of organic solids has been experimentally demonstrated, however no systematic theoretical analysis of this phenomenon has been carried out. We have conducted a theoretical study by using the Förster resonance energy transfer and Dexter carrier transfer mechanisms together with the Einstein-Smoluchowski diffusion equation to derive analytical models for the diffusion lengths (LD ) and diffusion coefficients (D) of singlet (S) and triplet (T) excitons in organic solids as functions of spectral overlap integral (J) , photoluminescence (PL) quantum yield (φD ) , dipole moment (μT ) and refractive index (n) of the photoactive material. The exciton diffusion lengths and diffusion coefficients in some selected organic solids were calculated, and we found that the singlet exciton diffusion length (LDS ) increases with φD and J, and decreases with n. Also, the triplet exciton diffusion length (LDT ) increases with φD and decreases with μT . These may be achieved through doping the organic solids into broad optical energy gap host materials as observed in previous experiments. The calculated exciton diffusion lengths are compared with experimental values and a reasonably good agreement is found between them. The results presented are expected to provide insight relevant to the synthesis of new organic solids for fabrication of bulk heterojunction organic solar cells characterized by better power conversion efficiency.

  11. The timing of cortical granule fusion, content dispersal, and endocytosis during fertilization of the hamster egg: an electrophysiological and histochemical study.

    Science.gov (United States)

    Kline, D; Stewart-Savage, J

    1994-03-01

    To determine the temporal relationship between cortical granule exocytosis and the repetitive calcium transients, which are characteristic of mammalian fertilization, we monitored membrane addition from exocytosis during fertilization of hamster eggs. Continuous measurement of membrane capacitance by applying a 3.1-nA alternating current at 375 Hz showed addition of cortical granule membrane. Simultaneous measurement of membrane potential revealed each calcium transient by the appearance of transient hyperpolarizing responses due to calcium-activated potassium channels in the egg. The initial membrane capacitance of the eggs averaged 736 +/- 44 pF (mean +/- SD; n = 7) and an increase in capacitance of 61 +/- 19 pF occurred within 4 sec of the start of the first hyperpolarizing response (HR) after fertilization. Immediately after the first increase in capacitance there was a gradual decline in membrane capacitance in all eggs and in five/seven eggs the capacitance returned to the unfertilized level in 7.8 +/- 4.4 min. The gradual decline in capacitance after the first increase indicated endocytosis, which was confirmed by the internalization of fluorescently labeled dextran. Superimposed on the gradual decline in membrane capacitance were smaller increases in capacitance that occurred with the second and later HRs. The total increase in capacitance from the first three events averaged 72 +/- 19 pF, representing an average increase in capacitance of about 10% of the capacitance of the unfertilized egg. By labeling eggs before and after permeabilization with two different fluorochromes attached to Lens culinaris agglutinin, we demonstrate that the dispersal of the cortical granules contents does not occur immediately after exocytosis. Our results demonstrate that cortical granule exocytosis in hamster eggs is closely coupled to the periodic increases in calcium, that the contents of the cortical granules are slow to disperse, and that after exocytosis, the surface

  12. Handling of bulk solids theory and practice

    CERN Document Server

    Shamlou, P A

    1990-01-01

    Handling of Bulk Solids provides a comprehensive discussion of the field of solids flow and handling in the process industries. Presentation of the subject follows classical lines of separate discussions for each topic, so each chapter is self-contained and can be read on its own. Topics discussed include bulk solids flow and handling properties; pressure profiles in bulk solids storage vessels; the design of storage silos for reliable discharge of bulk materials; gravity flow of particulate materials from storage vessels; pneumatic transportation of bulk solids; and the hazards of solid-mater

  13. Photoinduced Bulk Polarization and Its Effects on Photovoltaic Actions in Perovskite Solar Cells.

    Science.gov (United States)

    Wu, Ting; Collins, Liam; Zhang, Jia; Lin, Pei-Ying; Ahmadi, Mahshid; Jesse, Stephen; Hu, Bin

    2017-11-28

    This article reports an experimental demonstration of photoinduced bulk polarization in hysteresis-free methylammonium (MA) lead-halide perovskite solar cells [ITO/PEDOT:PSS/perovskite/PCBM/PEI/Ag]. An anomalous capacitance-voltage (CV) signal is observed as a broad "shoulder" in the depletion region from -0.5 to +0.5 V under photoexcitation based on CV measurements where a dc bias is gradually scanned to continuously drift mobile ions in order to detect local polarization under a low alternating bias (50 mV, 5 kHz). Essentially, gradually scanning the dc bias and applying a low alternating bias can separately generate continuously drifting ions and a bulk CV signal from local polarization under photoexcitation. Particularly, when the device efficiency is improved from 12.41% to 18.19% upon chlorine incorporation, this anomalous CV signal can be enhanced by a factor of 3. This anomalous CV signal can be assigned as the signature of photoinduced bulk polarization by distinguishing from surface polarization associated with interfacial charge accumulation. Meanwhile, replacing easy-rotational MA + with difficult-rotational formamidinium (FA + ) cations largely minimizes such anomalous CV signal, suggesting that photoinduced bulk polarization relies on the orientational freedom of dipolar organic cations. Furthermore, a Kelvin probe force microscopy study shows that chlorine incorporation can suppress the density of charged defects and thus enhances photoinduced bulk polarization due to the reduced screening effect from charged defects. A bias-dependent photoluminescence study indicates that increasing bulk polarization can suppress carrier recombination by decreasing charge capture probability through the Coulombic screening effect. Clearly, our studies provide an insightful understanding of photoinduced bulk polarization and its effects on photovoltaic actions in perovskite solar cells.

  14. Ferromagnetic bulk glassy alloys

    International Nuclear Information System (INIS)

    Inoue, Akihisa; Makino, Akihiro; Mizushima, Takao

    2000-01-01

    This paper deals with the review on the formation, thermal stability and magnetic properties of the Fe-based bulk glassy alloys in as-cast bulk and melt-spun ribbon forms. A large supercooled liquid region over 50 K before crystallization was obtained in Fe-(Al, Ga)-(P, C, B, Si), Fe-(Cr, Mo, Nb)-(Al, Ga)-(P, C, B) and (Fe, Co, Ni)-Zr-M-B (M=Ti, Hf, V, Nb, Ta, Cr, Mo and W) systems and bulk glassy alloys were produced in a thickness range below 2 mm for the Fe-(Al, Ga)-(P, C, B, Si) system and 6 mm for the Fe-Co-(Zr, Nb, Ta)-(Mo, W)-B system by copper-mold casting. The ring-shaped glassy Fe-(Al, Ga)-(P, C, B, Si) alloys exhibit much better soft magnetic properties as compared with the ring-shaped alloy made from the melt-spun ribbon because of the formation of the unique domain structure. The good combination of high glass-forming ability and good soft magnetic properties indicates the possibility of future development as a new bulk glassy magnetic material

  15. Time-dependent mobility and recombination of the photoinduced charge carriers in conjugated polymer/fullerene bulk heterojunction solar cells

    Science.gov (United States)

    Mozer, A. J.; Dennler, G.; Sariciftci, N. S.; Westerling, M.; Pivrikas, A.; Österbacka, R.; Juška, G.

    2005-07-01

    Time-dependent mobility and recombination in the blend of poly[2-methoxy-5-(3,7-dimethyloctyloxy)-phenylene vinylene] (MDMO-PPV) and 1-(3-methoxycarbonyl)propyl-1-phenyl-(6,6)- C61 (PCBM) is studied simultaneously using the photoinduced charge carrier extraction by linearly increasing voltage technique. The charge carriers are photogenerated by a strongly absorbed, 3 ns laser flash, and extracted by the application of a reverse bias voltage pulse after an adjustable delay time (tdel) . It is found that the mobility of the extracted charge carriers decreases with increasing delay time, especially shortly after photoexcitation. The time-dependent mobility μ(t) is attributed to the energy relaxation of the charge carriers towards the tail states of the density of states distribution. A model based on a dispersive bimolecular recombination is formulated, which properly describes the concentration decay of the extracted charge carriers at all measured temperatures and concentrations. The calculated bimolecular recombination coefficient β(t) is also found to be time-dependent exhibiting a power law dependence as β(t)=β0t-(1-γ) with increasing slope (1-γ) with decreasing temperatures. The temperature dependence study reveals that both the mobility and recombination of the photogenerated charge carriers are thermally activated processes with activation energy in the range of 0.1 eV. Finally, the direct comparison of μ(t) and β(t) shows that the recombination of the long-lived charge carriers is controlled by diffusion.

  16. Dynamic magnetoelectric effects in bulk and layered composites of cobalt zinc ferrite and lead zirconate titanate

    Energy Technology Data Exchange (ETDEWEB)

    Srinivasan, G.; Hayes, R.; DeVreugd, C.P. [Oakland University, Physics Department, Rochester, MI (United States); Laletsin, V.M.; Paddubnaya, N. [National Academy of Sciences, Institute of Technical Acoustics, Vitebsk (Belarus)

    2005-02-01

    Low-frequency magnetoelectric (ME) coupling is investigated in bulk samples and multilayers of cobalt zinc ferrite, Co{sub 1-x}Zn{sub x}Fe{sub 2}O{sub 4} (x=0-0.6), and lead zirconate titanate. In bulk samples, the transverse and longitudinal couplings are weak and of equal magnitude. A substantial strengthening of ME interactions is evident in layered structures, with the ME voltage coefficient a factor of 10-30 higher than in bulk samples. Important findings of our studies of layered composites are as follows. (i) The transverse coupling is stronger than the longitudinal coupling. (ii) The strength of ME interactions is dependent on Zn substitution, with a maximum for x=0.4. (iii) A weak coupling exists at the ferromagnetic-piezoelectric interface, as revealed by an analysis of the volume and static magnetic field dependence of ME voltage coefficients. (iv) The interface coupling k increases with Zn substitution and the k versus x profile shows a maximum centered at x=0.4. (v) The Zn-assisted enhancement can be attributed to efficient magneto-mechanical coupling in the ferrite. (orig.)

  17. Cosmology with bulk viscosity and the gravitino problem. Consequences of imperfect fluids on gravitino production

    Energy Technology Data Exchange (ETDEWEB)

    Buoninfante, L.; Lambiase, G. [Dipartimento di Fisica ' ' E.R. Caianiello' ' Universita di Salerno, Fisciano (Italy); INFN-Gruppo Collegato di Salerno, Fisciano (Italy)

    2017-05-15

    The gravitino problem is revisited in the framework of cosmological models in which the primordial cosmic matter is described by a relativistic imperfect fluid. Dissipative effects (or bulk viscosity effects) arise owing to the different cooling rates of the fluid components. We show that the effects of the bulk viscosity allow one to avoid the late abundance of gravitinos. In particular, for particular values of the parameters characterizing the cosmological model, the gravitino abundance turns out to be weakly depending on the reheating temperature. (orig.)

  18. Characterization of bulk and shear properties of basmati and non-basmati rice flour.

    Science.gov (United States)

    Jan, Shumaila; Ghoroi, Chinmay; Saxena, Dharmesh Chandra

    2018-01-01

    Flours are often unstable in relation to their flow performance, which is evident when a free-flowing material ceases to flow and the processing, handling, and production parameters depend on the inherent powder characteristics and their bulk behaviour. The present study was conducted to compare the flowability of basmati and non-basmati rice flour affecting bulk handling, which could be related to its particle size, shape and surface roughness (measured by atomic force microscopy) as well as bulk and shear properties, depending upon the processing conditions. Particle size (171.1-171.9 μm) of both samples was not significantly different. However, the flowability of the non-basmati rice flour was significantly affected by its particle shape (circularity 0.487), surface roughness (124.23 nm) and compressibility (25.32%) in comparison to basmati rice flour (circularity 0.653, surface roughness 113.59 nm and compressibility 21.09%), making it more cohesive than basmati rice flour. Also, basic flow energy was significantly higher in non-basmati flour, thus requiring more energy (147.54 mJ) to flow than basmati rice flour (130.15 mJ). Overall, flowability was analysed by applying three different pressures (3, 6 and 9 kPa), among which non-basmati rice flour was found to be less flowable (flow function coefficient (FFC) 2.33 at 9 kPa) in comparison to basmati (FFC 3.35 at 9 kPa), making bulk handling difficult. This study could be useful in designing processing equipment, hoppers and silos for rice flour handling. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  19. Controlling solution-phase polymer aggregation with molecular weight and solvent additives to optimize polymer-fullerene bulk heterojunction solar cells

    KAUST Repository

    Bartelt, Jonathan A.; Douglas, Jessica D.; Mateker, William R.; El Labban, Abdulrahman; Tassone, Christopher J.; Toney, Michael F.; Fré chet, Jean Mj J; Beaujuge, Pierre; McGehee, Michael D.

    2014-01-01

    The bulk heterojunction (BHJ) solar cell performance of many polymers depends on the polymer molecular weight (M n) and the solvent additive(s) used for solution processing. However, the mechanism that causes these dependencies is not well

  20. 29 CFR 794.137 - Effect of activities other than “wholesale or bulk distribution of petroleum products.”

    Science.gov (United States)

    2010-07-01

    ... distribution of petroleum products.â 794.137 Section 794.137 Labor Regulations Relating to Labor (Continued... RELATED TO REGULATIONS PARTIAL OVERTIME EXEMPTION FOR EMPLOYEES OF WHOLESALE OR BULK PETROLEUM... other than “wholesale or bulk distribution of petroleum products.” As previously noted, in some cases...

  1. Dose-rate effects on the bulk etch-rate of CR-39 track detector exposed to low-LET radiations

    CERN Document Server

    Yamauchi, T; Oda, K; Ikeda, T; Honda, Y; Tagawa, S

    1999-01-01

    The effect of gamma-rays and pulsed electrons has been investigated on the bulk etch rate of CR-39 detector at doses up to 100 kGy under various dose-rate between 0.0044 and 35.0 Gy/s. The bulk etch rate increased exponentially with the dose at every examined dose-rates. It was reveled to be strongly depend on the dose-rate: the bulk etch rate was decreased with increasing dose-rate at the same total dose. A primitive model was proposed to explain the dose-rate effect in which oxygen dissolved was assumed to dominate the damage formation process.

  2. Single pulsed-field magnetization on Gd-Ba-Cu-O Bulk HTS assembled for axial-gap type rotating machines

    International Nuclear Information System (INIS)

    Morita, E; Matsuzaki, H; Kimura, Y; Ohtani, I; Izumi, M; Nonaka, Y; Murakami, M; Ida, T; Sugimoto, H; Miki, M; Kitano, M

    2006-01-01

    We employed Gd-bulk HTS magnets as rotating poles for a smaller and lighter axial-gap type rotating machine. The bulk was placed between two vortex-type armature coils and cooled down to 77 K under zero-field. Pulsed current was applied to the vortex-type magnetizing coils. The trapped field distribution and transient flux behaviour strongly depend on the radial dimension of the armature vortex-type coil. In the present study, we show that there is an optimal radial dimension of magnetizing coils to the given bulk disk size to give a homogeneously conical distribution of the trapped flux

  3. 350 keV accelerator based PGNAA setup to detect nitrogen in bulk samples

    Energy Technology Data Exchange (ETDEWEB)

    Naqvi, A.A., E-mail: aanaqvi@kfupm.edu.sa [Department of Physics and King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Al-Matouq, Faris A.; Khiari, F.Z.; Gondal, M.A.; Rehman, Khateeb-ur [Department of Physics and King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Isab, A.A. [Department of Chemistry, King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia); Raashid, M.; Dastageer, M.A. [Department of Physics and King Fahd University of Petroleum and Minerals, Dhahran (Saudi Arabia)

    2013-11-21

    Nitrogen concentration was measured in explosive and narcotics proxy material, e.g. anthranilic acid, caffeine, melamine, and urea samples, bulk samples through thermal neutron capture reaction using 350 keV accelerator based prompt gamma ray neutron activation (PGNAA) setup. Intensity of 2.52, 3.53–3.68, 4.51, 5.27–5.30 and 10.38 MeV prompt gamma rays of nitrogen from the bulk samples was measured using a cylindrical 100 mm×100 mm (diameter×height ) BGO detector. Inspite of interference of nitrogen gamma rays from bulk samples with capture prompt gamma rays from BGO detector material, an excellent agreement between the experimental and calculated yields of nitrogen gamma rays has been obtained. This is an indication of the excellent performance of the PGNAA setup for detection of nitrogen in bulk samples.

  4. 350 keV accelerator based PGNAA setup to detect nitrogen in bulk samples

    International Nuclear Information System (INIS)

    Naqvi, A.A.; Al-Matouq, Faris A.; Khiari, F.Z.; Gondal, M.A.; Rehman, Khateeb-ur; Isab, A.A.; Raashid, M.; Dastageer, M.A.

    2013-01-01

    Nitrogen concentration was measured in explosive and narcotics proxy material, e.g. anthranilic acid, caffeine, melamine, and urea samples, bulk samples through thermal neutron capture reaction using 350 keV accelerator based prompt gamma ray neutron activation (PGNAA) setup. Intensity of 2.52, 3.53–3.68, 4.51, 5.27–5.30 and 10.38 MeV prompt gamma rays of nitrogen from the bulk samples was measured using a cylindrical 100 mm×100 mm (diameter×height ) BGO detector. Inspite of interference of nitrogen gamma rays from bulk samples with capture prompt gamma rays from BGO detector material, an excellent agreement between the experimental and calculated yields of nitrogen gamma rays has been obtained. This is an indication of the excellent performance of the PGNAA setup for detection of nitrogen in bulk samples

  5. 350 keV accelerator based PGNAA setup to detect nitrogen in bulk samples

    Science.gov (United States)

    Naqvi, A. A.; Al-Matouq, Faris A.; Khiari, F. Z.; Gondal, M. A.; Rehman, Khateeb-ur; Isab, A. A.; Raashid, M.; Dastageer, M. A.

    2013-11-01

    Nitrogen concentration was measured in explosive and narcotics proxy material, e.g. anthranilic acid, caffeine, melamine, and urea samples, bulk samples through thermal neutron capture reaction using 350 keV accelerator based prompt gamma ray neutron activation (PGNAA) setup. Intensity of 2.52, 3.53-3.68, 4.51, 5.27-5.30 and 10.38 MeV prompt gamma rays of nitrogen from the bulk samples was measured using a cylindrical 100 mm×100 mm (diameter×height ) BGO detector. Inspite of interference of nitrogen gamma rays from bulk samples with capture prompt gamma rays from BGO detector material, an excellent agreement between the experimental and calculated yields of nitrogen gamma rays has been obtained. This is an indication of the excellent performance of the PGNAA setup for detection of nitrogen in bulk samples.

  6. Als3 is a Candida albicans invasin that binds to cadherins and induces endocytosis by host cells.

    Directory of Open Access Journals (Sweden)

    Quynh T Phan

    2007-03-01

    Full Text Available Candida albicans is the most common cause of hematogenously disseminated and oropharyngeal candidiasis. Both of these diseases are characterized by fungal invasion of host cells. Previously, we have found that C. albicans hyphae invade endothelial cells and oral epithelial cells in vitro by inducing their own endocytosis. Therefore, we set out to identify the fungal surface protein and host cell receptors that mediate this process. We found that the C. albicans Als3 is required for the organism to be endocytosed by human umbilical vein endothelial cells and two different human oral epithelial lines. Affinity purification experiments with wild-type and an als3delta/als3delta mutant strain of C. albicans demonstrated that Als3 was required for C. albicans to bind to multiple host cell surface proteins, including N-cadherin on endothelial cells and E-cadherin on oral epithelial cells. Furthermore, latex beads coated with the recombinant N-terminal portion of Als3 were endocytosed by Chinese hamster ovary cells expressing human N-cadherin or E-cadherin, whereas control beads coated with bovine serum albumin were not. Molecular modeling of the interactions of the N-terminal region of Als3 with the ectodomains of N-cadherin and E-cadherin indicated that the binding parameters of Als3 to either cadherin are similar to those of cadherin-cadherin binding. Therefore, Als3 is a fungal invasin that mimics host cell cadherins and induces endocytosis by binding to N-cadherin on endothelial cells and E-cadherin on oral epithelial cells. These results uncover the first known fungal invasin and provide evidence that C. albicans Als3 is a molecular mimic of human cadherins.

  7. Bulk emission by higher-dimensional black holes: almost perfect blackbody radiation

    International Nuclear Information System (INIS)

    Hod, Shahar

    2011-01-01

    We study the Hawking radiation emitted into the bulk by (D + 1)-dimensional Schwarzschild black holes. It is well known that the black-hole spectrum departs from exact blackbody form due to the frequency dependence of the 'greybody' factors. For intermediate values of D (3 ≤ D ∼ > 1, the typical wavelengths in the black-hole spectrum are much shorter than the size of the black hole. In this regime, the greybody factors are well described by the geometric-optics approximation according to which they are almost frequency independent. Following this observation, we argue that for higher-dimensional black holes with D >> 1, the total power emitted into the bulk should be well approximated by the analytical formula for perfect blackbody radiation. We test the validity of this analytical prediction with numerical computations.

  8. Nerve growth factor enhances the CRE-dependent transcriptional activity activated by nobiletin in PC12 cells.

    Science.gov (United States)

    Takito, Jiro; Kimura, Junko; Kajima, Koji; Uozumi, Nobuyuki; Watanabe, Makoto; Yokosuka, Akihito; Mimaki, Yoshihiro; Nakamura, Masanori; Ohizumi, Yasushi

    2016-07-01

    Prevention and treatment of Alzheimer disease are urgent problems for elderly people in developed countries. We previously reported that nobiletin, a poly-methoxylated flavone from the citrus peel, improved the symptoms in various types of animal models of memory loss and activated the cAMP responsive element (CRE)-dependent transcription in PC12 cells. Nobiletin activated the cAMP/PKA/MEK/Erk/MAPK signaling pathway without using the TrkA signaling activated by nerve growth factor (NGF). Here, we examined the effect of combination of nobiletin and NGF on the CRE-dependent transcription in PC12 cells. Although NGF alone had little effect on the CRE-dependent transcription, NGF markedly enhanced the CRE-dependent transcription induced by nobiletin. The NGF-induced enhancement was neutralized by a TrkA antagonist, K252a. This effect of NGF was effective on the early signaling event elicited by nobiletin. These results suggested that there was crosstalk between NGF and nobiletin signaling in activating the CRE-dependent transcription in PC12 cells.

  9. Coupling brane fields to bulk supergravity

    Energy Technology Data Exchange (ETDEWEB)

    Parameswaran, Susha L. [Uppsala Univ. (Sweden). Theoretical Physics; Schmidt, Jonas [Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)

    2010-12-15

    In this note we present a simple, general prescription for coupling brane localized fields to bulk supergravity. We illustrate the procedure by considering 6D N=2 bulk supergravity on a 2D orbifold, with brane fields localized at the fixed points. The resulting action enjoys the full 6D N=2 symmetries in the bulk, and those of 4D N=1 supergravity at the brane positions. (orig.)

  10. Enhancement of magnetic coupling between permanent magnets and bulk superconductors through iron embedding

    International Nuclear Information System (INIS)

    Seki, H.; Kurabayashi, H.; Suzuki, A.; Ikeda, M.; Akiyama, S.; Murakami, M.

    2009-01-01

    Magnetic torque can be transferred without contact through the coupling of permanent magnets (PM) and bulk superconductors (BSC). For this purpose, permanent magnets should have multiple pole configuration like NSNS. The magnitude of the transferable torque depends on the field strength and the gap between PM and BSC. It was found that the torque decays quickly with the gap. In order to enhance the strength of transferable magnetic torque, we prepared bulk Y-Ba-Cu-O superconductors for which Fe bars are embedded. Holes about 1 mm in diameter were mechanically drilled into bulk Y-Ba-Cu-O, and Fe bars about 0.9 mm in diameter were inserted followed by impregnation of Bi-Pb-Sn alloys with low melting points. The composite of Y-Ba-Cu-O and Fe bars attract magnetic fields generated from permanent magnet before cooling, and thereby magnetic coupling will be improved. We have found that the magnetic torque force can be greatly enhanced through iron embedding.

  11. Lipoprotein-biomimetic nanostructure enables efficient targeting delivery of siRNA to Ras-activated glioblastoma cells via macropinocytosis

    Science.gov (United States)

    Huang, Jia-Lin; Jiang, Gan; Song, Qing-Xiang; Gu, Xiao; Hu, Meng; Wang, Xiao-Lin; Song, Hua-Hua; Chen, Le-Pei; Lin, Ying-Ying; Jiang, Di; Chen, Jun; Feng, Jun-Feng; Qiu, Yong-Ming; Jiang, Ji-Yao; Jiang, Xin-Guo; Chen, Hong-Zhuan; Gao, Xiao-Ling

    2017-05-01

    Hyperactivated Ras regulates many oncogenic pathways in several malignant human cancers including glioblastoma and it is an attractive target for cancer therapies. Ras activation in cancer cells drives protein internalization via macropinocytosis as a key nutrient-gaining process. By utilizing this unique endocytosis pathway, here we create a biologically inspired nanostructure that can induce cancer cells to `drink drugs' for targeting activating transcription factor-5 (ATF5), an overexpressed anti-apoptotic transcription factor in glioblastoma. Apolipoprotein E3-reconstituted high-density lipoprotein is used to encapsulate the siRNA-loaded calcium phosphate core and facilitate it to penetrate the blood-brain barrier, thus targeting the glioblastoma cells in a macropinocytosis-dependent manner. The nanostructure carrying ATF5 siRNA exerts remarkable RNA-interfering efficiency, increases glioblastoma cell apoptosis and inhibits tumour cell growth both in vitro and in xenograft tumour models. This strategy of targeting the macropinocytosis caused by Ras activation provides a nanoparticle-based approach for precision therapy in glioblastoma and other Ras-activated cancers.

  12. Elasticity of nanoparticles influences their blood circulation, phagocytosis, endocytosis, and targeting.

    Science.gov (United States)

    Anselmo, Aaron C; Zhang, Mengwen; Kumar, Sunny; Vogus, Douglas R; Menegatti, Stefano; Helgeson, Matthew E; Mitragotri, Samir

    2015-03-24

    The impact of physical and chemical modifications of nanoparticles on their biological function has been systemically investigated and exploited to improve their circulation and targeting. However, the impact of nanoparticles' flexibility (i.e., elastic modulus) on their function has been explored to a far lesser extent, and the potential benefits of tuning nanoparticle elasticity are not clear. Here, we describe a method to synthesize polyethylene glycol (PEG)-based hydrogel nanoparticles of uniform size (200 nm) with elastic moduli ranging from 0.255 to 3000 kPa. These particles are used to investigate the role of particle elasticity on key functions including blood circulation time, biodistribution, antibody-mediated targeting, endocytosis, and phagocytosis. Our results demonstrate that softer nanoparticles (10 kPa) offer enhanced circulation and subsequently enhanced targeting compared to harder nanoparticles (3000 kPa) in vivo. Furthermore, in vitro experiments show that softer nanoparticles exhibit significantly reduced cellular uptake in immune cells (J774 macrophages), endothelial cells (bEnd.3), and cancer cells (4T1). Tuning nanoparticle elasticity potentially offers a method to improve the biological fate of nanoparticles by offering enhanced circulation, reduced immune system uptake, and improved targeting.

  13. Activity-dependent branching ratios in stocks, solar x-ray flux, and the Bak-Tang-Wiesenfeld sandpile model.

    Science.gov (United States)

    Martin, Elliot; Shreim, Amer; Paczuski, Maya

    2010-01-01

    We define an activity-dependent branching ratio that allows comparison of different time series X(t). The branching ratio b(x) is defined as b(x)=E[xi(x)/x]. The random variable xi(x) is the value of the next signal given that the previous one is equal to x, so xi(x)=[X(t+1) | X(t)=x]. If b(x)>1, the process is on average supercritical when the signal is equal to x, while if b(x)efficient market hypothesis." For stock volumes, solar x-ray flux intensities, and the Bak-Tang-Wiesenfeld (BTW) sandpile model, b(x) is supercritical for small values of activity and subcritical for the largest ones, indicating a tendency to return to a typical value. For stock volumes this tendency has an approximate power-law behavior. For solar x-ray flux and the BTW model, there is a broad regime of activity where b(x) approximately equal 1, which we interpret as an indicator of critical behavior. This is true despite different underlying probability distributions for X(t) and for xi(x). For the BTW model the distribution of xi(x) is Gaussian, for x sufficiently larger than 1, and its variance grows linearly with x. Hence, the activity in the BTW model obeys a central limit theorem when sampling over past histories. The broad region of activity where b(x) is close to one disappears once bulk dissipation is introduced in the BTW model-supporting our hypothesis that it is an indicator of criticality.

  14. Pulsed-Field Magnetization Properties of Bulk Superconductors by Employment of Vortex-Type Coils

    Science.gov (United States)

    Deng, Z.; Shinohara, N.; Miki, M.; Felder, B.; Tsuzuki, K.; Watasaki, M.; Kawabe, S.; Taguchi, R.; Izumi, M.

    Vortex-type magnetizing coils are gaining more and more attention to activate bulk superconductors in pulsed-field magnetization (PFM) studies, compared with solenoid-type ones. Following existing reports, we present experimental results of the different penetration patterns of magnetic flux between the two