The structure and function of presynaptic endosomes

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Jähne, Sebastian, E-mail: sebastian.jaehne1@stud.uni-goettingen.de [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); International Max Planck Research School for Neurosciences, 37077 Göttingen (Germany); Rizzoli, Silvio O. [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); Helm, Martin S., E-mail: martin.helm@med.uni-goettingen.de [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); International Max Planck Research School for Molecular Biology, 37077 Göttingen (Germany)

2015-07-15

The function of endosomes and of endosome-like structures in the presynaptic compartment is still controversial. This is in part due to the absence of a consensus on definitions and markers for these compartments. Synaptic endosomes are sometimes seen as stable organelles, permanently present in the synapse. Alternatively, they are seen as short-lived intermediates in synaptic vesicle recycling, arising from the endocytosis of large vesicles from the plasma membrane, or from homotypic fusion of small vesicles. In addition, the potential function of the endosome is largely unknown in the synapse. Some groups have proposed that the endosome is involved in the sorting of synaptic vesicle proteins, albeit others have produced data that deny this possibility. In this review, we present the existing evidence for synaptic endosomes, we discuss their potential functions, and we highlight frequent technical pitfalls in the analysis of this elusive compartment. We also sketch a roadmap to definitely determine the role of synaptic endosomes for the synaptic vesicle cycle. Finally, we propose a common definition of synaptic endosome-like structures.

The structure and function of presynaptic endosomes

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Jähne, Sebastian; Rizzoli, Silvio O.; Helm, Martin S.

2015-01-01

The function of endosomes and of endosome-like structures in the presynaptic compartment is still controversial. This is in part due to the absence of a consensus on definitions and markers for these compartments. Synaptic endosomes are sometimes seen as stable organelles, permanently present in the synapse. Alternatively, they are seen as short-lived intermediates in synaptic vesicle recycling, arising from the endocytosis of large vesicles from the plasma membrane, or from homotypic fusion of small vesicles. In addition, the potential function of the endosome is largely unknown in the synapse. Some groups have proposed that the endosome is involved in the sorting of synaptic vesicle proteins, albeit others have produced data that deny this possibility. In this review, we present the existing evidence for synaptic endosomes, we discuss their potential functions, and we highlight frequent technical pitfalls in the analysis of this elusive compartment. We also sketch a roadmap to definitely determine the role of synaptic endosomes for the synaptic vesicle cycle. Finally, we propose a common definition of synaptic endosome-like structures

Membrane tethering complexes in the endosomal system

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Anne eSpang

2016-05-01

Full Text Available Vesicles that are generated by endocytic events at the plasma membrane are destined to early endosomes. A prerequisite for proper fusion is the tethering of two membrane entities. Tethering of vesicles to early endosomes is mediated by the CORVET complex, while fusion of late endosomes with lysosomes depends on the HOPS complex. Recycling through the TGN and to the plasma membrane is facilitated by the GARP and EARP complexes, respectively. However, there are other tethering functions in the endosomal system as there are multiple pathways through which proteins can be delivered from endosomes to either the TGN or the plasma membrane. Furthermore, complexes that may be part of novel tethering complexes have been recently identified. Thus it is likely that more tethering factors exist. In this review, I will provide an overview of different tethering complexes of the endosomal system and discuss how they may provide specificity in membrane traffic.

Differential endosomal sorting of a novel P2Y12 purinoreceptor mutant.

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Cunningham, Margaret R; Nisar, Shaista P; Cooke, Alexandra E; Emery, Elizabeth D; Mundell, Stuart J

2013-05-01

P2Y12 receptor internalization and recycling play an essential role in ADP-induced platelet activation. Recently, we identified a patient with a mild bleeding disorder carrying a heterozygous mutation of P2Y12 (P341A) whose P2Y12 receptor recycling was significantly compromised. Using human cell line models, we identified key proteins regulating wild-type (WT) P2Y12 recycling and investigated P2Y12 -P341A receptor traffic. Treatment with ADP resulted in delayed Rab5-dependent internalization of P341A when compared with WT P2Y12 . While WT P2Y12 rapidly recycled back to the membrane via Rab4 and Rab11 recycling pathways, limited P341A recycling was observed, which relied upon Rab11 activity. Although minimal receptor degradation was evident, P341A was localized in Rab7-positive endosomes with considerable agonist-dependent accumulation in the trans-Golgi network (TGN). Rab7 activity is known to facilitate recruitment of retromer complex proteins to endosomes to transport cargo to the TGN. Here, we identified that P341A colocalized with Vps26; depletion of which blocked limited recycling and promoted receptor degradation. This study has identified key points of divergence in the endocytic traffic of P341A versus WT-P2Y12 . Given that these pathways are retained in human platelets, this research helps define the molecular mechanisms regulating P2Y12 receptor traffic and explain the compromised receptor function in the platelets of the P2Y12 -P341A-expressing patient. © 2013 John Wiley & Sons A/S.

Synaptic activity regulates AMPA receptor trafficking through different recycling pathways

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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

Molecular assemblies and membrane domains in multivesicular endosome dynamics

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Falguieres, Thomas; Luyet, Pierre-Philippe; Gruenberg, Jean

2009-01-01

Along the degradation pathway, endosomes exhibit a characteristic multivesicular organization, resulting from the budding of vesicles into the endosomal lumen. After endocytosis and transport to early endosomes, activated signaling receptors are incorporated into these intralumenal vesicles through the action of the ESCRT machinery, a process that contributes to terminate signaling. Then, the vesicles and their protein cargo are further transported towards lysosomes for degradation. Evidence also shows that intralumenal vesicles can undergo 'back-fusion' with the late endosome limiting membrane, a route exploited by some pathogens and presumably followed by proteins and lipids that need to be recycled from within the endosomal lumen. This process depends on the late endosomal lipid lysobisphosphatidic acid and its putative effector Alix/AIP1, and is presumably coupled to the invagination of the endosomal limiting membrane at the molecular level via ESCRT proteins. In this review, we discuss the intra-endosomal transport routes in mammalian cells, and in particular the different mechanisms involved in membrane invagination, vesicle formation and fusion in a space inaccessible to proteins known to control intracellular membrane traffic.

Yersinia pestis Targets the Host Endosome Recycling Pathway during the Biogenesis of the Yersinia-Containing Vacuole To Avoid Killing by Macrophages

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Connor, Michael G.; Pulsifer, Amanda R.; Ceresa, Brian K.

2018-01-01

ABSTRACT Yersinia pestis has evolved many strategies to evade the innate immune system. One of these strategies is the ability to survive within macrophages. Upon phagocytosis, Y. pestis prevents phagolysosome maturation and establishes a modified compartment termed the Yersinia-containing vacuole (YCV). Y. pestis actively inhibits the acidification of this compartment, and eventually, the YCV transitions from a tight-fitting vacuole into a spacious replicative vacuole. The mechanisms to generate the YCV have not been defined. However, we hypothesized that YCV biogenesis requires Y. pestis interactions with specific host factors to subvert normal vesicular trafficking. In order to identify these factors, we performed a genome-wide RNA interference (RNAi) screen to identify host factors required for Y. pestis survival in macrophages. This screen revealed that 71 host proteins are required for intracellular survival of Y. pestis. Of particular interest was the enrichment for genes involved in endosome recycling. Moreover, we demonstrated that Y. pestis actively recruits Rab4a and Rab11b to the YCV in a type three secretion system-independent manner, indicating remodeling of the YCV by Y. pestis to resemble a recycling endosome. While recruitment of Rab4a was necessary to inhibit YCV acidification and lysosomal fusion early during infection, Rab11b appeared to contribute to later stages of YCV biogenesis. We also discovered that Y. pestis disrupts global host endocytic recycling in macrophages, possibly through sequestration of Rab11b, and this process is required for bacterial replication. These data provide the first evidence that Y. pestis targets the host endocytic recycling pathway to avoid phagolysosomal maturation and generate the YCV. PMID:29463656

Actin-Sorting Nexin 27 (SNX27)-Retromer Complex Mediates Rapid Parathyroid Hormone Receptor Recycling*

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McGarvey, Jennifer C.; Xiao, Kunhong; Bowman, Shanna L.; Mamonova, Tatyana; Zhang, Qiangmin; Bisello, Alessandro; Sneddon, W. Bruce; Ardura, Juan A.; Jean-Alphonse, Frederic; Vilardaga, Jean-Pierre; Puthenveedu, Manojkumar A.; Friedman, Peter A.

2016-01-01

The G protein-coupled parathyroid hormone receptor (PTHR) regulates mineral-ion homeostasis and bone remodeling. Upon parathyroid hormone (PTH) stimulation, the PTHR internalizes into early endosomes and subsequently traffics to the retromer complex, a sorting platform on early endosomes that promotes recycling of surface receptors. The C terminus of the PTHR contains a type I PDZ ligand that binds PDZ domain-containing proteins. Mass spectrometry identified sorting nexin 27 (SNX27) in isolated endosomes as a PTHR binding partner. PTH treatment enriched endosomal PTHR. SNX27 contains a PDZ domain and serves as a cargo selector for the retromer complex. VPS26, VPS29, and VPS35 retromer subunits were isolated with PTHR in endosomes from cells stimulated with PTH. Molecular dynamics and protein binding studies establish that PTHR and SNX27 interactions depend on the PDZ recognition motif in PTHR and the PDZ domain of SNX27. Depletion of either SNX27 or VPS35 or actin depolymerization decreased the rate of PTHR recycling following agonist stimulation. Mutating the PDZ ligand of PTHR abolished the interaction with SNX27 but did not affect the overall rate of recycling, suggesting that PTHR may directly engage the retromer complex. Coimmunoprecipitation and overlay experiments show that both intact and mutated PTHR bind retromer through the VPS26 protomer and sequentially assemble a ternary complex with PTHR and SNX27. SNX27-independent recycling may involve N-ethylmaleimide-sensitive factor, which binds both PDZ intact and mutant PTHRs. We conclude that PTHR recycles rapidly through at least two pathways, one involving the ASRT complex of actin, SNX27, and retromer and another possibly involving N-ethylmaleimide-sensitive factor. PMID:27008860

The functional interplay of Rab11, FIP3 and Rho proteins on the endosomal recycling pathway controls cell shape and symmetry.

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Bouchet, Jérôme; McCaffrey, Mary W; Graziani, Andrea; Alcover, Andrés

2018-07-04

Several families of small GTPases regulate a variety of fundamental cellular processes, encompassing growth factor signal transduction, vesicular trafficking and control of the cytoskeleton. Frequently, their action is hierarchical and complementary, but much of the detail of their functional interactions remains to be clarified. It is well established that Rab family members regulate a variety of intracellular vesicle trafficking pathways. Moreover, Rho family GTPases are pivotal for the control of the actin and microtubule cytoskeleton. However, the interplay between these 2 types of GTPases has been rarely reported. We discuss here our recent findings showing that Rab11, a key regulator of endosomal recycling, and Rac1, a central actin cytoskeleton regulator involved in lamellipodium formation and cell migration, interplay on endosomes through the Rab11 effector FIP3. In the context of the rapidly reactive T lymphocytes, Rab11-Rac1 endosomal functional interplay is important to control cell shape changes and cell symmetry during lymphocyte spreading and immunological synapse formation and ultimately modulate T cell activation.

Anterograde trafficking of KCa3.1 in polarized epithelia is Rab1- and Rab8-dependent and recycling endosome-independent.

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Claudia A Bertuccio

Full Text Available The intermediate conductance, Ca2+-activated K+ channel (KCa3.1 targets to the basolateral (BL membrane in polarized epithelia where it plays a key role in transepithelial ion transport. However, there are no studies defining the anterograde and retrograde trafficking of KCa3.1 in polarized epithelia. Herein, we utilize Biotin Ligase Acceptor Peptide (BLAP-tagged KCa3.1 to address these trafficking steps in polarized epithelia, using MDCK, Caco-2 and FRT cells. We demonstrate that KCa3.1 is exclusively targeted to the BL membrane in these cells when grown on filter supports. Following endocytosis, KCa3.1 degradation is prevented by inhibition of lysosomal/proteosomal pathways. Further, the ubiquitylation of KCa3.1 is increased following endocytosis from the BL membrane and PR-619, a deubiquitylase inhibitor, prevents degradation, indicating KCa3.1 is targeted for degradation by ubiquitylation. We demonstrate that KCa3.1 is targeted to the BL membrane in polarized LLC-PK1 cells which lack the μ1B subunit of the AP-1 complex, indicating BL targeting of KCa3.1 is independent of μ1B. As Rabs 1, 2, 6 and 8 play roles in ER/Golgi exit and trafficking of proteins to the BL membrane, we evaluated the role of these Rabs in the trafficking of KCa3.1. In the presence of dominant negative Rab1 or Rab8, KCa3.1 cell surface expression was significantly reduced, whereas Rabs 2 and 6 had no effect. We also co-immunoprecipitated KCa3.1 with both Rab1 and Rab8. These results suggest these Rabs are necessary for the anterograde trafficking of KCa3.1. Finally, we determined whether KCa3.1 traffics directly to the BL membrane or through recycling endosomes in MDCK cells. For these studies, we used either recycling endosome ablation or dominant negative RME-1 constructs and determined that KCa3.1 is trafficked directly to the BL membrane rather than via recycling endosomes. These results are the first to describe the anterograde and retrograde trafficking of KCa3

Human ClC-6 is a late endosomal glycoprotein that associates with detergent-resistant lipid domains.

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Sofie Ignoul

Full Text Available BACKGROUND: The mammalian CLC protein family comprises nine members (ClC-1 to -7 and ClC-Ka, -Kb that function either as plasma membrane chloride channels or as intracellular chloride/proton antiporters, and that sustain a broad spectrum of cellular processes, such as membrane excitability, transepithelial transport, endocytosis and lysosomal degradation. In this study we focus on human ClC-6, which is structurally most related to the late endosomal/lysomal ClC-7. PRINCIPAL FINDINGS: Using a polyclonal affinity-purified antibody directed against a unique epitope in the ClC-6 COOH-terminal tail, we show that human ClC-6, when transfected in COS-1 cells, is N-glycosylated in a region that is evolutionary poorly conserved between mammalian CLC proteins and that is located between the predicted helices K and M. Three asparagine residues (N410, N422 and N432 have been defined by mutagenesis as acceptor sites for N-glycosylation, but only two of the three sites seem to be simultaneously N-glycosylated. In a differentiated human neuroblastoma cell line (SH-SY5Y, endogenous ClC-6 colocalizes with LAMP-1, a late endosomal/lysosomal marker, but not with early/recycling endosomal markers such as EEA-1 and transferrin receptor. In contrast, when transiently expressed in COS-1 or HeLa cells, human ClC-6 mainly overlaps with markers for early/recycling endosomes (transferrin receptor, EEA-1, Rab5, Rab4 and not with late endosomal/lysosomal markers (LAMP-1, Rab7. Analogously, overexpression of human ClC-6 in SH-SY5Y cells also leads to an early/recycling endosomal localization of the exogenously expressed ClC-6 protein. Finally, in transiently transfected COS-1 cells, ClC-6 copurifies with detergent-resistant membrane fractions, suggesting its partitioning in lipid rafts. Mutating a juxtamembrane string of basic amino acids (amino acids 71-75: KKGRR disturbs the association with detergent-resistant membrane fractions and also affects the segregation of ClC-6

A TOCA/CDC-42/PAR/WAVE functional module required for retrograde endocytic recycling

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Bai, Zhiyong; Grant, Barth D.

2015-01-01

Endosome-to-Golgi transport is required for the function of many key membrane proteins and lipids, including signaling receptors, small-molecule transporters, and adhesion proteins. The retromer complex is well-known for its role in cargo sorting and vesicle budding from early endosomes, in most cases leading to cargo fusion with the trans-Golgi network (TGN). Transport from recycling endosomes to the TGN has also been reported, but much less is understood about the molecules that mediate this transport step. Here we provide evidence that the F-BAR domain proteins TOCA-1 and TOCA-2 (Transducer of Cdc42 dependent actin assembly), the small GTPase CDC-42 (Cell division control protein 42), associated polarity proteins PAR-6 (Partitioning defective 6) and PKC-3/atypical protein kinase C, and the WAVE actin nucleation complex mediate the transport of MIG-14/Wls and TGN-38/TGN38 cargo proteins from the recycling endosome to the TGN in Caenorhabditis elegans. Our results indicate that CDC-42, the TOCA proteins, and the WAVE component WVE-1 are enriched on RME-1–positive recycling endosomes in the intestine, unlike retromer components that act on early endosomes. Furthermore, we find that retrograde cargo TGN-38 is trapped in early endosomes after depletion of SNX-3 (a retromer component) but is mainly trapped in recycling endosomes after depletion of CDC-42, indicating that the CDC-42–associated complex functions after retromer in a distinct organelle. Thus, we identify a group of interacting proteins that mediate retrograde recycling, and link these proteins to a poorly understood trafficking step, recycling endosome-to-Golgi transport. We also provide evidence for the physiological importance of this pathway in WNT signaling. PMID:25775511

A Role for EHD4 in the Regulation of Early Endosomal Transport

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Sharma, Mahak; Naslavsky, Naava; Caplan, Steve

2009-01-01

All four of the C-terminal Eps15 homology domain (EHD) proteins have been implicated in the regulation of endocytic trafficking. However, the high level of amino acid sequence identity among these proteins has made it challenging to elucidate the precise function of individual EHD proteins. We demonstrate here with specific peptide antibodies that endogenous EHD4 localizes to Rab5-, early embryonic antigen 1 (EEA1)- and Arf6-containing endosomes and colocalizes with internalized transferrin in the cell periphery. Knock-down of EHD4 expression by both small interfering RNA and short hairpin RNA leads to the generation of enlarged early endosomal structures that contain Rab5 and EEA1 as well as internalized transferrin or major histocompatibility complex class I molecules. In addition, cargo destined for degradation, such as internalized low-density lipoprotein, also accumulates in the enlarged early endosomes in EHD4-depleted cells. Moreover, we have demonstrated that these enlarged early endosomes are enriched in levels of the activated GTP-bound Rab5. Finally, we show that endogenous EHD4 and EHD1 interact in cells, suggesting coordinated involvement in the regulation of receptor transport along the early endosome to endocytic recycling compartment axis. The results presented herein provide evidence that EHD4 is involved in the control of trafficking at the early endosome and regulates exit of cargo toward both the recycling compartment and the late endocytic pathway. PMID:18331452

Cholesterol Regulates Syntaxin 6 Trafficking at trans-Golgi Network Endosomal Boundaries

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Meritxell Reverter

2014-05-01

Full Text Available Inhibition of cholesterol export from late endosomes causes cellular cholesterol imbalance, including cholesterol depletion in the trans-Golgi network (TGN. Here, using Chinese hamster ovary (CHO Niemann-Pick type C1 (NPC1 mutant cell lines and human NPC1 mutant fibroblasts, we show that altered cholesterol levels at the TGN/endosome boundaries trigger Syntaxin 6 (Stx6 accumulation into VAMP3, transferrin, and Rab11-positive recycling endosomes (REs. This increases Stx6/VAMP3 interaction and interferes with the recycling of αVβ3 and α5β1 integrins and cell migration, possibly in a Stx6-dependent manner. In NPC1 mutant cells, restoration of cholesterol levels in the TGN, but not inhibition of VAMP3, restores the steady-state localization of Stx6 in the TGN. Furthermore, elevation of RE cholesterol is associated with increased amounts of Stx6 in RE. Hence, the fine-tuning of cholesterol levels at the TGN-RE boundaries together with a subset of cholesterol-sensitive SNARE proteins may play a regulatory role in cell migration and invasion.

Apical transport of influenza A virus ribonucleoprotein requires Rab11-positive recycling endosome.

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Fumitaka Momose

Full Text Available Influenza A virus RNA genome exists as eight-segmented ribonucleoprotein complexes containing viral RNA polymerase and nucleoprotein (vRNPs. Packaging of vRNPs and virus budding take place at the apical plasma membrane (APM. However, little is known about the molecular mechanisms of apical transport of newly synthesized vRNP. Transfection of fluorescent-labeled antibody and subsequent live cell imaging revealed that punctate vRNP signals moved along microtubules rapidly but intermittently in both directions, suggestive of vesicle trafficking. Using a series of Rab family protein, we demonstrated that progeny vRNP localized to recycling endosome (RE in an active/GTP-bound Rab11-dependent manner. The vRNP interacted with Rab11 through viral RNA polymerase. The localization of vRNP to RE and subsequent accumulation to the APM were impaired by overexpression of Rab binding domains (RBD of Rab11 family interacting proteins (Rab11-FIPs. Similarly, no APM accumulation was observed by overexpression of class II Rab11-FIP mutants lacking RBD. These results suggest that the progeny vRNP makes use of Rab11-dependent RE machinery for APM trafficking.

Rab9-dependent retrograde transport and endosomal sorting of the endopeptidase furin

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Chia, Pei Zhi Cheryl; Gasnereau, Isabelle; Lieu, Zi Zhao; Gleeson, Paul A.

2011-01-01

The endopeptidase furin and the trans-Golgi network protein TGN38 are membrane proteins that recycle between the TGN and plasma membrane. TGN38 is transported by a retromer-dependent pathway from early endosomes to the TGN, whereas the intracellular transport of furin is poorly defined. Here we have identified the itinerary and transport requirements of furin. Using internalisation assays, we show that furin transits the early and late endosomes en route to the TGN. The GTPase Rab9 and the TGN golgin GCC185, components of the late endosome-to-TGN pathway, were required for efficient TGN retrieval of furin. By contrast, TGN38 trafficking was independent of Rab9 and GCC185. To identify the sorting signals for the early endosome-to-TGN pathway, the trafficking of furin–TGN38 chimeras was investigated. The diversion of furin from the Rab9-dependent late-endosome-to-TGN pathway to the retromer-dependent early-endosome-to-TGN pathway required both the transmembrane domain and cytoplasmic tail of TGN38. We present evidence to suggest that the length of the transmembrane domain is a contributing factor in endosomal sorting. Overall, these data show that furin uses the Rab9-dependent pathway from late endosomes and that retrograde transport directly from early endosomes is dependent on both the transmembrane domain and the cytoplasmic tail. PMID:21693586

The Endosome Localized Arf-GAP AGAP1 Modulates Dendritic Spine Morphology Downstream of the Neurodevelopmental Disorder Factor Dysbindin

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Miranda Arnold

2016-09-01

Full Text Available AGAP1 is an Arf1 GTPase activating protein that interacts with the vesicle-associated protein complexes adaptor protein 3 (AP-3 and Biogenesis of Lysosome Related Organelles Complex-1 (BLOC-1. Overexpression of AGAP1 in non-neuronal cells results in an accumulation of endosomal cargoes, which suggests a role in endosome-dependent traffic. In addition, AGAP1 is a candidate susceptibility gene for two neurodevelopmental disorders, autism spectrum disorder (ASD and schizophrenia (SZ; yet its localization and function in neurons have not been described. Here, we describe that AGAP1 localizes to axons, dendrites, dendritic spines, and synapses, colocalizing preferentially with markers of early and recycling endosomes. Functional studies reveal overexpression and down-regulation of AGAP1 affects both neuronal endosomal trafficking and dendritic spine morphology, supporting a role for AGAP1 in the recycling endosomal trafficking involved in their morphogenesis. Finally, we determined the sensitivity of AGAP1 expression to mutations in the DTNBP1 gene, which is associated with neurodevelopmental disorder, and found that AGAP1 mRNA and protein levels are selectively reduced in the null allele of the mouse orthologue of DTNBP1. We postulate that endosomal trafficking contributes to the pathogenesis of neurodevelopmental disorders affecting dendritic spine morphology, and thus excitatory synapse structure and function.

Antigen processing and remodeling of the endosomal pathway: requirements for antigen cross-presentation.

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Ewoud Bernardus Compeer

2012-03-01

Full Text Available The cross-presentation of endocytosed antigen as peptide/class I MHC complexes plays a central role in the elicitation of CD8+ T cell clones that mediate anti-viral and anti-tumor immune responses. While it has been clear that there are specific subsets of professional antigen presenting cells (APC capable of antigen cross-presentation, description of mechanisms involved is still ongoing. Especially amongst dendritic cells (DC, there are specialized subsets that are highly proficient at antigen cross-presentation. We here present a focused survey on the cell biological processes in the endosomal pathway that support antigen cross-presentation. This review highlight DC-intrinsic mechanisms that facilitate the cross-presentation of endocytosed antigen, including receptor-mediated uptake, recycling and maturation including the sorting of membrane proteins, dynamic remodeling of endosomal structures and cell-surface directed endosomal trafficking. We will conclude with description of pathogen-induced deviation of endosomal processing, and discuss how immune evasion strategies pertaining endosomal trafficking may preclude antigen cross-presentation.

GGA3 mediates TrkA endocytic recycling to promote sustained Akt phosphorylation and cell survival

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Li, Xuezhi; Lavigne, Pierre; Lavoie, Christine

2015-01-01

Although TrkA postendocytic sorting significantly influences neuronal cell survival and differentiation, the molecular mechanism underlying TrkA receptor sorting in the recycling or degradation pathways remains poorly understood. Here we demonstrate that Golgi-localized, γ adaptin-ear–containing ADP ribosylation factor-binding protein 3 (GGA3) interacts directly with the TrkA cytoplasmic tail through an internal DXXLL motif and mediates the functional recycling of TrkA to the plasma membrane. We find that GGA3 depletion by siRNA delays TrkA recycling, accelerates TrkA degradation, attenuates sustained NGF-induced Akt activation, and reduces cell survival. We also show that GGA3’s effect on TrkA recycling is dependent on the activation of Arf6. This work identifies GGA3 as a key player in a novel DXXLL-mediated endosomal sorting machinery that targets TrkA to the plasma membrane, where it prolongs the activation of Akt signaling and survival responses. PMID:26446845

Heterotrimeric G protein subunits are located on rat liver endosomes

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Van Dyke Rebecca W

2004-01-01

Full Text Available Abstract Background Rat liver endosomes contain activated insulin receptors and downstream signal transduction molecules. We undertook these studies to determine whether endosomes also contain heterotrimeric G proteins that may be involved in signal transduction from G protein-coupled receptors. Results By Western blotting Gsα, Giα1,2, Giα3 and Gβ were enriched in both canalicular (CM and basolateral (BLM membranes but also readily detectable on three types of purified rat liver endosomes in the order recycling receptor compartment (RRC > compartment for uncoupling of receptor and ligand (CURL > multivesicular bodies (MVB >> purified secondary lysosomes. Western blotting with antibodies to Na, K-ATPase and to other proteins associated with plasma membranes and intracellular organelles indicated this was not due to contamination of endosome preparations by CM or BLM. Adenylate cyclase (AC was also identified on purified CM, BLM, RRC, CURL and MVB. Percoll gradient fractionation of liver postnuclear supernatants demonstrated co-occurrence of endosomes and heterotrimeric G protein subunits in fractions with little plasma membrane markers. By confocal microscopy, punctate staining for Gsα, Giα3 and Gβ corresponded to punctate areas of endocytosed Texas red-dextran in hepatocytes from control and cholera toxin-treated livers. Conclusion We conclude that heterotrimeric G protein subunits as well as AC likely traffic into hepatocytes on endosome membranes, possibly generating downstream signals spatially separate from signalling generated at the plasma membrane, analogous to the role(s of internalized insulin receptors.

IL4/PGE2 induction of an enlarged early endosomal compartment in mouse macrophages is Rab5-dependent

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Wainszelbaum, Marisa J.; Proctor, Brandon M.; Pontow, Suzanne E.; Stahl, Philip D.; Barbieri, M. Alejandro

2006-01-01

The endosomal compartment and the plasma membrane form a complex partnership that controls signal transduction and trafficking of different molecules. The specificity and functionality of the early endocytic pathway are regulated by a growing number of Rab GTPases, particularly Rab5. In this study, we demonstrate that IL4 (a Th-2 cytokine) and prostaglandin E 2 (PGE 2 ) synergistically induce Rab5 and several Rab effector proteins, including Rin1 and EEA1, and promote the formation of an enlarged early endocytic (EEE) compartment. Endosome enlargement is linked to a substantial induction of the mannose receptor (MR), a well-characterized macrophage endocytic receptor. Both MR levels and MR-mediated endocytosis are enhanced approximately 7-fold. Fluid-phase endocytosis is also elevated in treated cells. Light microscopy and fractionation studies reveal that MR colocalizes predominantly with Rab5a and partially with Rab11, an endosomal recycling pathway marker. Using retroviral expression of Rab5a:S34N, a dominant negative mutant, and siRNA Rab5a silencing, we demonstrate that Rab5a is essential for the large endosome phenotype and for localization of MR in these structures. We speculate that the EEE is maintained by activated Rab5, and that the EEE phenotype is part of some macrophage developmental program such as cell fusion, a characteristic of IL4-stimulated cells

L-Type Voltage-Gated Ca2+ Channels Regulate Synaptic-Activity-Triggered Recycling Endosome Fusion in Neuronal Dendrites

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Brian G. Hiester

2017-11-01

Full Text Available The repertoire and abundance of proteins displayed on the surface of neuronal dendrites are tuned by regulated fusion of recycling endosomes (REs with the dendritic plasma membrane. While this process is critical for neuronal function and plasticity, how synaptic activity drives RE fusion remains unexplored. We demonstrate a multistep fusion mechanism that requires Ca2+ from distinct sources. NMDA receptor Ca2+ initiates RE fusion with the plasma membrane, while L-type voltage-gated Ca2+ channels (L-VGCCs regulate whether fused REs collapse into the membrane or reform without transferring their cargo to the cell surface. Accordingly, NMDA receptor activation triggered AMPA-type glutamate receptor trafficking to the dendritic surface in an L-VGCC-dependent manner. Conversely, potentiating L-VGCCs enhanced AMPA receptor surface expression only when NMDA receptors were also active. Thus L-VGCCs play a role in tuning activity-triggered surface expression of key synaptic proteins by gating the mode of RE fusion.

Enhanced Endosomal Escape by Light-Fueled Liquid-Metal Transformer.

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Lu, Yue; Lin, Yiliang; Chen, Zhaowei; Hu, Quanyin; Liu, Yang; Yu, Shuangjiang; Gao, Wei; Dickey, Michael D; Gu, Zhen

2017-04-12

Effective endosomal escape remains as the "holy grail" for endocytosis-based intracellular drug delivery. To date, most of the endosomal escape strategies rely on small molecules, cationic polymers, or pore-forming proteins, which are often limited by the systemic toxicity and lack of specificity. We describe here a light-fueled liquid-metal transformer for effective endosomal escape-facilitated cargo delivery via a chemical-mechanical process. The nanoscale transformer can be prepared by a simple approach of sonicating a low-toxicity liquid-metal. When coated with graphene quantum dots (GQDs), the resulting nanospheres demonstrate the ability to absorb and convert photoenergy to drive the simultaneous phase separation and morphological transformation of the inner liquid-metal core. The morphological transformation from nanospheres to hollow nanorods with a remarkable change of aspect ratio can physically disrupt the endosomal membrane to promote endosomal escape of payloads. This metal-based nanotransformer equipped with GQDs provides a new strategy for facilitating effective endosomal escape to achieve spatiotemporally controlled drug delivery with enhanced efficacy.

Post-Golgi anterograde transport requires GARP-dependent endosome-to-TGN retrograde transport

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Hirata, Tetsuya; Fujita, Morihisa; Nakamura, Shota; Gotoh, Kazuyoshi; Motooka, Daisuke; Murakami, Yoshiko; Maeda, Yusuke; Kinoshita, Taroh

2015-01-01

The importance of endosome-to–trans-Golgi network (TGN) retrograde transport in the anterograde transport of proteins is unclear. In this study, genome-wide screening of the factors necessary for efficient anterograde protein transport in human haploid cells identified subunits of the Golgi-associated retrograde protein (GARP) complex, a tethering factor involved in endosome-to-TGN transport. Knockout (KO) of each of the four GARP subunits, VPS51–VPS54, in HEK293 cells caused severely defective anterograde transport of both glycosylphosphatidylinositol (GPI)-anchored and transmembrane proteins from the TGN. Overexpression of VAMP4, v-SNARE, in VPS54-KO cells partially restored not only endosome-to-TGN retrograde transport, but also anterograde transport of both GPI-anchored and transmembrane proteins. Further screening for genes whose overexpression normalized the VPS54-KO phenotype identified TMEM87A, encoding an uncharacterized Golgi-resident membrane protein. Overexpression of TMEM87A or its close homologue TMEM87B in VPS54-KO cells partially restored endosome-to-TGN retrograde transport and anterograde transport. Therefore GARP- and VAMP4-dependent endosome-to-TGN retrograde transport is required for recycling of molecules critical for efficient post-Golgi anterograde transport of cell-surface integral membrane proteins. In addition, TMEM87A and TMEM87B are involved in endosome-to-TGN retrograde transport. PMID:26157166

Endosome-based protein trafficking and Ca2+ homeostasis in the heart

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Jerry eCurran

2015-02-01

Full Text Available The ability to dynamically regulate, traffic, retain, and recycle proteins within the cell membrane is fundamental to life and central to the normal function of the heart and cardiovascular system. In the heart, these systems are essential for the regulation of cardiac calcium, both at the level of the plasma membrane, but also at local domains of the endoplasmic reticulum, sarcoplasmic reticulum, mitochondria, nucleus, and nuclear envelope. One intracellular pathway often overlooked in relation to cardiovascular calcium regulation and signaling is the endosome-based trafficking pathway. Highlighting its importance, this system and its molecular components are evolutionarily conserved across all metazoans. However, remarkably little is known of how endosome-based protein trafficking and recycling functions within mammalian cells systems, especially in the heart. The vast majority of what is known has been derived from heterologous cell systems. However, recently, more appropriate cell and animal models been developed that have allowed researchers to begin to understand how this system functions within the intact physiological environment. All excitable cells, including cardiomyocytes, depend on the proper expression and organization of multiple ion channels, pumps, exchangers, and transporters within the plasma membrane. As the endosomal system acts to regulate the expression and localization of membrane proteins, understanding the in vivo function of this system in the heart is important. This review will focus on endosome-based protein trafficking in the heart in both health and disease. Special emphasis will be given to the role played by the family of endocytic regulatory proteins, C-terminal Eps15 homology domain -containing proteins (EHDs, as recent data demonstrates that this family of proteins is essential for the proper trafficking and localization and of key proteins involved in excitation-contraction coupling.

Specific recycling receptors are targeted to the immune synapse by the intraflagellar transport system

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Finetti, Francesca; Patrussi, Laura; Masi, Giulia; Onnis, Anna; Galgano, Donatella; Lucherini, Orso Maria; Pazour, Gregory J.; Baldari, Cosima T.

2014-01-01

ABSTRACT T cell activation requires sustained signaling at the immune synapse, a specialized interface with the antigen-presenting cell (APC) that assembles following T cell antigen receptor (TCR) engagement by major histocompatibility complex (MHC)-bound peptide. Central to sustained signaling is the continuous recruitment of TCRs to the immune synapse. These TCRs are partly mobilized from an endosomal pool by polarized recycling. We have identified IFT20, a component of the intraflagellar transport (IFT) system that controls ciliogenesis, as a central regulator of TCR recycling to the immune synapse. Here, we have investigated the interplay of IFT20 with the Rab GTPase network that controls recycling. We found that IFT20 forms a complex with Rab5 and the TCR on early endosomes. IFT20 knockdown (IFT20KD) resulted in a block in the recycling pathway, leading to a build-up of recycling TCRs in Rab5+ endosomes. Recycling of the transferrin receptor (TfR), but not of CXCR4, was disrupted by IFT20 deficiency. The IFT components IFT52 and IFT57 were found to act together with IFT20 to regulate TCR and TfR recycling. The results provide novel insights into the mechanisms that control TCR recycling and immune synapse assembly, and underscore the trafficking-related function of the IFT system beyond ciliogenesis. PMID:24554435

Recycling Mentors: an intergenerational, service-learning program to promote recycling and environmental awareness.

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D'abundo, Michelle L; Fugate-Whitlock, Elizabeth I; Fiala, Kelly A

2011-01-01

The purpose of Recycling Mentors was to implement an intergenerational, service-learning program focused on promoting recycling and environmental awareness among students enrolled in Community Health (HEA 301) and Current Issues in Gerontology (GRN 440/540) and adults older than 60 years. Recycling Mentors was conducted in New Hanover County (NHC), North Carolina, where a moderate climate and coastal location attracts many tourists, retirees, and college students. A community like NHC is a good place to implement service-learning that educates both students and older adults about the benefits of recycling to individual health and the environment. During the Fall 2009 semester, undergraduate and graduate students completed institutional review board training and then conducted the program with older adults. The education component of Recycling Mentors included a pre/post survey, brochure, and scheduled visits. Overall, Recycling Mentors was positive service-learning experience with students identifying salient outcomes such as learning about recycling and the environment and working with older adults. In addition, teaching the education component of Recycling Mentors was good practice for students who will be the future health professionals. While service-learning and environmentally themed projects are common, a program that combines the 2 like Recycling Mentors is unique and has the potential to motivate individual change while positively impacting the local community and the environment.

dOCRL maintains immune cell quiescence by regulating endosomal traffic.

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Steven J Del Signore

2017-10-01

Full Text Available Lowe Syndrome is a developmental disorder characterized by eye, kidney, and neurological pathologies, and is caused by mutations in the phosphatidylinositol-5-phosphatase OCRL. OCRL plays diverse roles in endocytic and endolysosomal trafficking, cytokinesis, and ciliogenesis, but it is unclear which of these cellular functions underlie specific patient symptoms. Here, we show that mutation of Drosophila OCRL causes cell-autonomous activation of hemocytes, which are macrophage-like cells of the innate immune system. Among many cell biological defects that we identified in docrl mutant hemocytes, we pinpointed the cause of innate immune cell activation to reduced Rab11-dependent recycling traffic and concomitantly increased Rab7-dependent late endosome traffic. Loss of docrl amplifies multiple immune-relevant signals, including Toll, Jun kinase, and STAT, and leads to Rab11-sensitive mis-sorting and excessive secretion of the Toll ligand Spåtzle. Thus, docrl regulation of endosomal traffic maintains hemocytes in a poised, but quiescent state, suggesting mechanisms by which endosomal misregulation of signaling may contribute to symptoms of Lowe syndrome.

Huntingtin coordinates the dynein-mediated dynamic positioning of endosomes and lysosomes

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Caviston, Juliane P.; Zajac, Allison L.; Tokito, Mariko; Holzbaur, Erika L.F.

2011-01-01

Huntingtin (Htt) is a membrane-associated scaffolding protein that interacts with microtubule motors as well as actin-associated adaptor molecules. We examined a role for Htt in the dynein-mediated intracellular trafficking of endosomes and lysosomes. In HeLa cells depleted of either Htt or dynein, early, recycling, and late endosomes (LE)/lysosomes all become dispersed. Despite altered organelle localization, kinetic assays indicate only minor defects in intracellular trafficking. Expression of full-length Htt is required to restore organelle localization in Htt-depleted cells, supporting a role for Htt as a scaffold that promotes functional interactions along its length. In dynein-depleted cells, LE/lysosomes accumulate in tight patches near the cortex, apparently enmeshed by cortactin-positive actin filaments; Latrunculin B-treatment disperses these patches. Peripheral LE/lysosomes in dynein-depleted cells no longer colocalize with microtubules. Htt may be required for this off-loading, as the loss of microtubule association is not seen in Htt-depleted cells or in cells depleted of both dynein and Htt. Inhibition of kinesin-1 relocalizes peripheral LE/lysosomes induced by Htt depletion but not by dynein depletion, consistent with their detachment from microtubules upon dynein knockdown. Together, these data support a model of Htt as a facilitator of dynein-mediated trafficking that may regulate the cytoskeletal association of dynamic organelles. PMID:21169558

The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation.

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Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

2016-07-22

The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation*

Science.gov (United States)

Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

2016-01-01

The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. PMID:27302062

Effect of diphtheria toxin T-domain on endosomal pH

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A. J. Labyntsev

2015-08-01

Full Text Available A key step in the mode of cytotoxic action of diphtheria toxin (DT is the transfer of its catalytic domain (Cd from endosomes into the cytosol. The main activity in this process is performed by the transport domain (Td, but the molecular mechanism of its action remains unknown. We have previously shown that Td can have some influence on the endosomal transport of DT. The aim of this work was to study the effect of diphtheria toxin on the toxin compartmentalization in the intracellular transporting pathway and endosomal pH. We used recombinant fragments of DT, which differed only by the presence of Td in their structure, fused with fluorescent proteins. It was shown that the toxin fragment with Td moved slower by the pathway early-late endosomes-lysosomes, and had a slightly different pattern of colocalization with endosomal markers than DT fragment without Td. In addition, endosomes containing DT fragments with Td had a constant pH of about 6.5 from the 10th to 50th minute of observation, for the same time endosomes containing DT fragments without Td demons­trated a decrease in pH from 6.3 to 5.5. These results indicate that Td inhibits acidification of endosomal medium. One of possible explanations for this may be the effect of the ion channel formed by the T-domain on the process of the endosomal acidification. This property of Td may not only inhibit maturation of endosomes but also inhibit activation of endosomal pH-dependent proteases, and this promotes successful transport of Cd into the cell cytosol.

Late endosomal cholesterol accumulation leads to impaired intra-endosomal trafficking.

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Komla Sobo

Full Text Available BACKGROUND: Pathological accumulation of cholesterol in late endosomes is observed in lysosomal storage diseases such as Niemann-Pick type C. We here analyzed the effects of cholesterol accumulation in NPC cells, or as phenocopied by the drug U18666A, on late endosomes membrane organization and dynamics. METHODOLOGY/PRINCIPAL FINDINGS: Cholesterol accumulation did not lead to an increase in the raft to non-raft membrane ratio as anticipated. Strikingly, we observed a 2-3 fold increase in the size of the compartment. Most importantly, properties and dynamics of late endosomal intralumenal vesicles were altered as revealed by reduced late endosomal vacuolation induced by the mutant pore-forming toxin ASSP, reduced intoxication by the anthrax lethal toxin and inhibition of infection by the Vesicular Stomatitis Virus. CONCLUSIONS/SIGNIFICANCE: These results suggest that back fusion of intralumenal vesicles with the limiting membrane of late endosomes is dramatically perturbed upon cholesterol accumulation.

Chimeric forms of furin and TGN38 are transported with the plasma membrane in the trans-Golgi network via distinct endosomal pathways.

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Mallet, W G; Maxfield, F R

1999-07-26

Furin and TGN38 are menbrane proteins that cycle between the plasma membrane and the trans-Golgi network (TGN), each maintaining a predominant distribution in the TGN. We have used chimeric proteins with an extracellular Tac domain and the cytoplasmic domain of TGN38 or furin to study the trafficking of these proteins in endosomes. Previously, we demonstrated that the postendocytic trafficking of Tac-TGN38 to the TGN is via the endocytic recycling pathway (Ghosh, R.N.,W.G. Mallet,T.T. Soe,T.E.McGraw, and F.R. Maxfield.1998.J. Cell Biol.142:923-936). Here we show that internalized Tac-furin is delivered to the TGN through late endosomes, bypassing the endocytic recycling compartment. The transport of Tac-furin from late endosomes to the TGN appears to proceed via an efficient, single-pass mechanism. Delivery of Tac-furin but not Tac-TGN38 to the TGN is blocked by nocodazole, and the two pathways are also differentially affected by wortmannin. These studies demonstrate the existence of two independentpathways for endosomal transport of proteins to the TGN from the plasma membrane.

Recycling domains in plant cell morphogenesis: small GTPase effectors, plasma membrane signalling and the exocyst.

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Zárský, Viktor; Potocký, Martin

2010-04-01

The Rho/Rop small GTPase regulatory module is central for initiating exocytotically ACDs (active cortical domains) in plant cell cortex, and a growing array of Rop regulators and effectors are being discovered in plants. Structural membrane phospholipids are important constituents of cells as well as signals, and phospholipid-modifying enzymes are well known effectors of small GTPases. We have shown that PLDs (phospholipases D) and their product, PA (phosphatidic acid), belong to the regulators of the secretory pathway in plants. We have also shown that specific NOXs (NADPH oxidases) producing ROS (reactive oxygen species) are involved in cell growth as exemplified by pollen tubes and root hairs. Most plant cells exhibit several distinct plasma membrane domains (ACDs), established and maintained by endocytosis/exocytosis-driven membrane protein recycling. We proposed recently the concept of a 'recycling domain' (RD), uniting the ACD and the connected endosomal recycling compartment (endosome), as a dynamic spatiotemporal entity. We have described a putative GTPase-effector complex exocyst involved in exocytic vesicle tethering in plants. Owing to the multiplicity of its Exo70 subunits, this complex, along with many RabA GTPases (putative recycling endosome organizers), may belong to core regulators of RD organization in plants.

The sorting protein PACS-2 promotes ErbB signalling by regulating recycling of the metalloproteinase ADAM17

DEFF Research Database (Denmark)

Dombernowsky, Sarah Louise; Samsøe-Petersen, Jacob; Petersen, Camilla Hansson

2015-01-01

The metalloproteinase ADAM17 activates ErbB signalling by releasing ligands from the cell surface, a key step underlying epithelial development, growth and tumour progression. However, mechanisms acutely controlling ADAM17 cell-surface availability to modulate the extent of ErbB ligand release....... PACS-2 co-localizes with ADAM17 on early endosomes and PACS-2 knockdown decreases the recycling and stability of internalized ADAM17. Hence, PACS-2 sustains ADAM17 cell-surface activity by diverting ADAM17 away from degradative pathways. Interestingly, Pacs2-deficient mice display significantly reduced...

Recycling Endosomes and Viral Infection.

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Vale-Costa, Sílvia; Amorim, Maria João

2016-03-08

Many viruses exploit specific arms of the endomembrane system. The unique composition of each arm prompts the development of remarkably specific interactions between viruses and sub-organelles. This review focuses on the viral-host interactions occurring on the endocytic recycling compartment (ERC), and mediated by its regulatory Ras-related in brain (Rab) GTPase Rab11. This protein regulates trafficking from the ERC and the trans-Golgi network to the plasma membrane. Such transport comprises intricate networks of proteins/lipids operating sequentially from the membrane of origin up to the cell surface. Rab11 is also emerging as a critical factor in an increasing number of infections by major animal viruses, including pathogens that provoke human disease. Understanding the interplay between the ERC and viruses is a milestone in human health. Rab11 has been associated with several steps of the viral lifecycles by unclear processes that use sophisticated diversified host machinery. For this reason, we first explore the state-of-the-art on processes regulating membrane composition and trafficking. Subsequently, this review outlines viral interactions with the ERC, highlighting current knowledge on viral-host binding partners. Finally, using examples from the few mechanistic studies available we emphasize how ERC functions are adjusted during infection to remodel cytoskeleton dynamics, innate immunity and membrane composition.

Reduce--recycle--reuse: guidelines for promoting perioperative waste management.

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Laustsen, Gary

2007-04-01

The perioperative environment generates large amounts of waste, which negatively affects local and global ecosystems. To manage this waste health care facility leaders must focus on identifying correctable issues, work with relevant stakeholders to promote solutions, and adopt systematic procedural changes. Nurses and managers can moderate negative environmental effects by promoting reduction, recycling, and reuse of materials in the perioperative setting.

Neuron specific Rab4 effector GRASP-1 coordinates membrane specialization and maturation of recycling endosomes

NARCIS (Netherlands)

C.C. Hoogenraad (Casper); I. Popa (Ioana); K. Futai (Kensuke); E. Sanchez-Martinez (Emma); P. Wulf (Phebe); T. van Vlijmen (Thijs); B.R. Dortland (Bjorn); V. Oorschot (Viola); R. Govers (Robert); M. Monti (Maria); A.J.R. Heck (Albert); M. Sheng (Morgan); J. Klumperman (Judith); H. Rehmann (Holger); D. Jaarsma (Dick); L.C. Kapitein (Lukas); P. van der Sluijs

2010-01-01

textabstractThe endosomal pathway in neuronal dendrites is essential for membrane receptor trafficking and proper synaptic function and plasticity. However, the molecular mechanisms that organize specific endocytic trafficking routes are poorly understood. Here, we identify GRIP-associated protein-1

Two barcodes encoded by the type-1 PDZ and by phospho-Ser312 regulate retromer/WASH-mediated sorting of the ß1-adrenergic receptor from endosomes to the plasma membrane.

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Nooh, Mohammed M; Bahouth, Suleiman W

2017-01-01

Recycling of the majority of agonist-internalized GPCR is dependent on a type I-PDZ "barcode" in their C-tail. The recycling of wild-type (WT) ß 1 -AR is also dependent on its default "type-1 PDZ barcode", but trafficking of the ß 1 -AR is inhibited when PKA or its substrate serine at position 312 (Ser 312 ) are inactivated. We tested the hypothesis that phospho-Ser 312 provided a second barcode for ß 1 -AR sorting from endosomes to the plasma membrane by determining the role of retromer/WASH complexes in ß 1 -AR trafficking. Recycling of WT ß 1 -AR or WT ß 2 -AR was dependent on targeting the retromer to endosomal membranes via SNX3 and rab7a, and on complexing the retromer to the WASH pentamer via the C-tail of FAM21 (FAM21 C ). These maneuvers however, did not inhibit the recycling of a phospho-Ser 312 ß 1 -AR mimic ((S312D) ß 1 -AR). Knockdown of the trans-acting PDZ protein sorting nexin27 (SNX27) inhibited the recycling of WT ß 1 -AR and WT ß 2 -AR, but had no effect on (S312D) ß 1 -AR∆PDZ or on phosphorylation of WT ß 1 -AR by PKA at Ser 312 . However, depletion of FKBP15, a FAM21 C -binding endosomal protein, selectively inhibited WT ß 1 -AR but not ß 2 -AR recycling, suggesting divergence might exist in GPCR trafficking roadmaps. These results indicate that two barcodes are involved in sorting WT ß 1 -AR out of early endosomes. The first and antecedent "barcode" was the "type-1 PDZ", followed by a second reversible "phospho-Ser 312 " verification "barcode". This organization allows tight regulation of ß 1 -AR density to signaling intensity in conditions associated with aberrant ß 1 -AR signaling such as in hypertension and heart failure. Copyright © 2016 Elsevier Inc. All rights reserved.

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

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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.

Retrogradely Transported TrkA Endosomes Signal Locally within Dendrites to Maintain Sympathetic Neuron Synapses

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Kathryn M. Lehigh

2017-04-01

Full Text Available Sympathetic neurons require NGF from their target fields for survival, axonal target innervation, dendritic growth and formation, and maintenance of synaptic inputs from preganglionic neurons. Target-derived NGF signals are propagated retrogradely, from distal axons to somata of sympathetic neurons via TrkA signaling endosomes. We report that a subset of TrkA endosomes that are transported from distal axons to cell bodies translocate into dendrites, where they are signaling competent and move bidirectionally, in close proximity to synaptic protein clusters. Using a strategy for spatially confined inhibition of TrkA kinase activity, we found that distal-axon-derived TrkA signaling endosomes are necessary within sympathetic neuron dendrites for maintenance of synapses. Thus, TrkA signaling endosomes have unique functions in different cellular compartments. Moreover, target-derived NGF mediates circuit formation and synapse maintenance through TrkA endosome signaling within dendrites to promote aggregation of postsynaptic protein complexes.

Adiponectin release and insulin receptor targeting share trans-Golgi-dependent endosomal trafficking routes

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Maria Rödiger

2018-02-01

Full Text Available Objective: Intracellular vesicle trafficking maintains cellular structures and functions. The assembly of cargo-laden vesicles at the trans-Golgi network is initiated by the ARF family of small GTPases. Here, we demonstrate the role of the trans-Golgi localized monomeric GTPase ARFRP1 in endosomal-mediated vesicle trafficking of mature adipocytes. Methods: Control (Arfrp1flox/flox and inducible fat-specific Arfrp1 knockout (Arfrp1iAT−/− mice were metabolically characterized. In vitro experiments on mature 3T3-L1 cells and primary mouse adipocytes were conducted to validate the impact of ARFRP1 on localization of adiponectin and the insulin receptor. Finally, secretion and transferrin-based uptake and recycling assays were performed with HeLa and HeLa M-C1 cells. Results: We identified the ARFRP1-based sorting machinery to be involved in vesicle trafficking relying on the endosomal compartment for cell surface delivery. Secretion of adiponectin from fat depots was selectively reduced in Arfrp1iAT−/− mice, and Arfrp1-depleted 3T3-L1 adipocytes revealed an accumulation of adiponectin in Rab11-positive endosomes. Plasma adiponectin deficiency of Arfrp1iAT−/− mice resulted in deteriorated hepatic insulin sensitivity, increased gluconeogenesis and elevated fasting blood glucose levels. Additionally, the insulin receptor, undergoing endocytic recycling after ligand binding, was less abundant at the plasma membrane of adipocytes lacking Arfrp1. This had detrimental effects on adipose insulin signaling, followed by insufficient suppression of basal lipolytic activity and impaired adipose tissue expansion. Conclusions: Our findings suggest that adiponectin secretion and insulin receptor surface targeting utilize the same post-Golgi trafficking pathways that are essential for an appropriate systemic insulin sensitivity and glucose homeostasis. Keywords: Adiponectin, ARFRP1, Exocytosis, Insulin receptor, trans-Golgi

Internal structure of magnetic endosomes

Science.gov (United States)

Rivière, C.; Wilhelm, C.; Cousin, F.; Dupuis, V.; Gazeau, F.; Perzynski, R.

2007-01-01

The internal structure of biological vesicles filled with magnetic nanoparticles is investigated using the following complementary analyses: electronic transmission microscopy, dynamic probing by magneto-optical birefringence and structural probing by Small Angle Neutron Scattering (SANS). These magnetic vesicles are magnetic endosomes obtained via a non-specific interaction between cells and anionic magnetic iron oxide nanoparticles. Thanks to a magnetic purification process, they are probed at two different stages of their formation within HeLa cells: (i) adsorption of nanoparticles onto the cellular membrane and (ii) their subsequent internalisation within endosomes. Differences in the microenvironment of the magnetic nanoparticles at those two different stages are highlighted here. The dynamics of magnetic nanoparticles adsorbed onto cellular membranes and confined within endosomes is respectively 3 and 5 orders of magnitude slower than for isolated magnetic nanoparticles in aqueous media. Interestingly, SANS experiments show that magnetic endosomes have an internal structure close to decorated vesicles, with magnetic nanoparticles locally decorating the endosome membrane, inside their inner-sphere. These results, important for future biomedical applications, suggest that multiple fusions of decorated vesicles are the biological processes underlying the endocytosis of that kind of nanometric materials.

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

Overexpression of Rab22a hampers the transport between endosomes and the Golgi apparatus

International Nuclear Information System (INIS)

Mesa, Rosana; Magadan, Javier; Barbieri, Alejandro; Lopez, Cecilia; Stahl, Philip D.; Mayorga, Luis S.

2005-01-01

The transport and sorting of soluble and membrane-associated macromolecules arriving at endosomal compartments require a complex set of Rab proteins. Rab22a has been localized to the endocytic compartment; however, very little is known about the function of Rab22a and inconsistent results have been reported in studies performed in different cell lines. To characterize the function of Rab22a in endocytic transport, the wild-type protein (Rab22a WT), a hydrolysis-deficient mutant (Rab22a Q64L), and a mutant with reduced affinity for GTP (Rab22a S19N) were expressed in CHO cells. None of the three Rab22a constructs affected the transport of rhodamine-dextran to lysosomes, the digestion of internalized proteins, or the lysosomal localization of cathepsin D. In contrast with the mild effect of Rab22a on the endosome-lysosome route, cells expressing Rab22a WT and Rab22a Q64L presented a strong delay in the retrograde transport of cholera toxin from endosomes to the Golgi apparatus. Moreover, these cells accumulated the cation independent mannose 6-phosphate receptor in endosomes. These observations indicate that Rab22a can affect the trafficking from endosomes to the Golgi apparatus probably by promoting fusion among endosomes and impairing the proper segregation of membrane domains required for targeting to the trans-Golgi network (TGN)

Early to Late Endosome Trafficking Controls Secretion and Zymogen Activation in Rodent and Human Pancreatic Acinar Cells.

Science.gov (United States)

Messenger, Scott W; Thomas, Diana Dh; Cooley, Michelle M; Jones, Elaina K; Falkowski, Michelle A; August, Benjamin K; Fernandez, Luis A; Gorelick, Fred S; Groblewski, Guy E

2015-11-01

Pancreatic acinar cells have an expanded apical endosomal system, the physiological and pathophysiological significance of which is still emerging. Phosphatidylinositol-3,5-bisphosphate (PI(3,5)P 2 ) is an essential phospholipid generated by PIKfyve, which phosphorylates phosphatidylinositol-3-phosphate (PI(3)P). PI(3,5)P 2 is necessary for maturation of early endosomes (EE) to late endosomes (LE). Inhibition of EE to LE trafficking enhances anterograde endosomal trafficking and secretion at the plasma membrane by default through a recycling endosome (RE) intermediate. We assessed the effects of modulating PIKfyve activity on apical trafficking and pancreatitis responses in pancreatic acinar cells. Inhibition of EE to LE trafficking was achieved using pharmacological inhibitors of PIKfyve, expression of dominant negative PIKfyve K1877E, or constitutively active Rab5-GTP Q79L. Anterograde endosomal trafficking was manipulated by expression of constitutively active and dominant negative Rab11a mutants. The effects of these agents on secretion, endolysosomal exocytosis of lysosome associated membrane protein (LAMP1), and trypsinogen activation in response to high-dose CCK-8, bile acids and cigarette toxin was determined. PIKfyve inhibition increased basal and stimulated secretion. Adenoviral overexpression of PIKfyve decreased secretion leading to cellular death. Expression of Rab5-GTP Q79L or Rab11a-GTP Q70L enhanced secretion. Conversely, dominant-negative Rab11a-GDP S25N reduced secretion. High-dose CCK inhibited endolysosomal exocytosis that was reversed by PIKfyve inhibition. PIKfyve inhibition blocked intracellular trypsin accumulation and cellular damage responses to high CCK-8, tobacco toxin, and bile salts in both rodent and human acini. These data demonstrate that EE-LE trafficking acutely controls acinar secretion and the intracellular activation of zymogens leading to the pathogenicity of acute pancreatitis.

Effectiveness of Recycling Promotion and Education Initiatives among First-Generation Ethnic Minorities in Ontario, Canada

Directory of Open Access Journals (Sweden)

Calvin Lakhan

2016-05-01

Full Text Available This study examines how first-generation ethnic minorities respond to different types of recycling promotion and education campaigns (P&E used by municipalities in Ontario, Canada. A total of eight focus group sessions were conducted over an eight-week period to gauge participant attitudes and responses towards print (newspaper and signs and electronic (websites P&E messaging. Participants were asked to comment on message “recognition”, “clarity”, “the ability to increase recycling awareness” and “the ability to affect changes in recycling behavior”. Results from the focus group sessions suggest that none of the P&E mediums tested were able to increase recycling awareness or change recycling behavior in any meaningful way. First-generation ethnic minorities struggle with recognizing the central theme and purpose of P&E advertisements. Respondents also found existing campaigns excessively complex and confusing, and were not familiar with many of the terms and symbols used in existing P&E messaging. Other findings suggest that ethnic minorities are skeptical and distrustful of the municipalities’ intentions with respect to what they do with the waste after it is collected. The findings from this study lead to the recommendation that municipalities rethink and redesign recycling promotion and education initiatives to better engage minority communities.

Gαs regulates Glucagon-Like Peptide 1 Receptor-mediated cyclic AMP generation at Rab5 endosomal compartment.

Science.gov (United States)

Girada, Shravan Babu; Kuna, Ramya S; Bele, Shilpak; Zhu, Zhimeng; Chakravarthi, N R; DiMarchi, Richard D; Mitra, Prasenjit

2017-10-01

Upon activation, G protein coupled receptors (GPCRs) associate with heterotrimeric G proteins at the plasma membrane to initiate second messenger signaling. Subsequently, the activated receptor experiences desensitization, internalization, and recycling back to the plasma membrane, or it undergoes lysosomal degradation. Recent reports highlight specific cases of persistent cyclic AMP generation by internalized GPCRs, although the functional significance and mechanistic details remain to be defined. Cyclic AMP generation from internalized Glucagon-Like Peptide-1 Receptor (GLP-1R) has previously been reported from our laboratory. This study aimed at deciphering the molecular mechanism by which internalized GLP-R supports sustained cyclic AMP generation upon receptor activation in pancreatic beta cells. We studied the time course of cyclic AMP generation following GLP-1R activation with particular emphasis on defining the location where cyclic AMP is generated. Detection involved a novel GLP-1 conjugate coupled with immunofluorescence using specific endosomal markers. Finally, we employed co-immunoprecipitation as well as immunofluorescence to assess the protein-protein interactions that regulate GLP-1R mediated cyclic AMP generation at endosomes. Our data reveal that prolonged association of G protein α subunit Gαs with activated GLP-1R contributed to sustained cyclic AMP generation at Rab 5 endosomal compartment. The findings provide the mechanism of endosomal cyclic AMP generation following GLP-1R activation. We identified the specific compartment that serves as an organizing center to generate endosomal cyclic AMP by internalized activated receptor complex. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

The PDZ protein GIPC regulates trafficking of the LPA1 receptor from APPL signaling endosomes and attenuates the cell's response to LPA.

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Tal Varsano

Full Text Available Lysophosphatidic acid (LPA mediates diverse cellular responses through the activation of at least six LPA receptors--LPA(1-6, but the interacting proteins and signaling pathways that mediate the specificity of these receptors are largely unknown. We noticed that LPA(1 contains a PDZ binding motif (SVV identical to that present in two other proteins that interact with the PDZ protein GIPC. GIPC is involved in endocytic trafficking of several receptors including TrkA, VEGFR2, lutropin and dopamine D2 receptors. Here we show that GIPC binds directly to the PDZ binding motif of LPA(1 but not that of other LPA receptors. LPA(1 colocalizes and coimmunoprecipitates with GIPC and its binding partner APPL, an activator of Akt signaling found on APPL signaling endosomes. GIPC depletion by siRNA disturbed trafficking of LPA(1 to EEA1 early endosomes and promoted LPA(1 mediated Akt signaling, cell proliferation, and cell motility. We propose that GIPC binds LPA(1 and promotes its trafficking from APPL-containing signaling endosomes to EEA1 early endosomes and thus attenuates LPA-mediated Akt signaling from APPL endosomes.

Biomechanics and Thermodynamics of Nanoparticle Interactions with Plasma and Endosomal Membrane Lipids in Cellular Uptake and Endosomal Escape

Science.gov (United States)

2015-01-01

To be effective for cytoplasmic delivery of therapeutics, nanoparticles (NPs) taken up via endocytic pathways must efficiently transport across the cell membrane and subsequently escape from the secondary endosomes. We hypothesized that the biomechanical and thermodynamic interactions of NPs with plasma and endosomal membrane lipids are involved in these processes. Using model plasma and endosomal lipid membranes, we compared the interactions of cationic NPs composed of poly(d,l-lactide-co-glycolide) modified with the dichain surfactant didodecyldimethylammonium bromide (DMAB) or the single-chain surfactant cetyltrimethylammonium bromide (CTAB) vs anionic unmodified NPs of similar size. We validated our hypothesis in doxorubicin-sensitive (MCF-7, with relatively fluid membranes) and resistant breast cancer cells (MCF-7/ADR, with rigid membranes). Despite their cationic surface charges, DMAB- and CTAB-modified NPs showed different patterns of biophysical interaction: DMAB-modified NPs induced bending of the model plasma membrane, whereas CTAB-modified NPs condensed the membrane, thereby resisted bending. Unmodified NPs showed no effects on bending. DMAB-modified NPs also induced thermodynamic instability of the model endosomal membrane, whereas CTAB-modified and unmodified NPs had no effect. Since bending of the plasma membrane and destabilization of the endosomal membrane are critical biophysical processes in NP cellular uptake and endosomal escape, respectively, we tested these NPs for cellular uptake and drug efficacy. Confocal imaging showed that in both sensitive and resistant cells DMAB-modified NPs exhibited greater cellular uptake and escape from endosomes than CTAB-modified or unmodified NPs. Further, paclitaxel-loaded DMAB-modified NPs induced greater cytotoxicity even in resistant cells than CTAB-modified or unmodified NPs or drug in solution, demonstrating the potential of DMAB-modified NPs to overcome the transport barrier in resistant cells. In

Early to Late Endosome Trafficking Controls Secretion and Zymogen Activation in Rodent and Human Pancreatic Acinar CellsSummary

Directory of Open Access Journals (Sweden)

Scott W. Messenger

2015-11-01

Full Text Available Background & Aims: Pancreatic acinar cells have an expanded apical endosomal system, the physiologic and pathophysiologic significance of which is still emerging. Phosphatidylinositol-3,5-bisphosphate [PI(3,5P2] is an essential phospholipid generated by phosphatidylinositol 3-phosphate 5-kinase (PIKfyve, which phosphorylates phosphatidylinositol-3-phosphate (PI3P. PI(3,5P2 is necessary for maturation of early endosomes (EE to late endosomes (LE. Inhibition of EE to LE trafficking enhances anterograde endosomal trafficking and secretion at the plasma membrane by default through a recycling endosome (RE intermediate. We assessed the effects of modulating PIKfyve activity on apical trafficking and pancreatitis responses in pancreatic acinar cells. Methods: Inhibition of EE to LE trafficking was achieved using pharmacologic inhibitors of PIKfyve, expression of dominant negative PIKfyve K1877E, or constitutively active Rab5-GTP Q79L. Anterograde endosomal trafficking was manipulated by expression of constitutively active and dominant negative Rab11a mutants. The effects of these agents on secretion, endolysosomal exocytosis of lysosome associated membrane protein (LAMP1, and trypsinogen activation in response to supramaximal cholecystokinin (CCK-8, bile acids, and cigarette toxin was determined. Results: PIKfyve inhibition increased basal and stimulated secretion. Adenoviral overexpression of PIKfyve decreased secretion leading to cellular death. Expression of Rab5-GTP Q79L or Rab11a-GTP Q70L enhanced secretion. Conversely, dominant-negative Rab11a-GDP S25N reduced secretion. High-dose CCK inhibited endolysosomal exocytosis that was reversed by PIKfyve inhibition. PIKfyve inhibition blocked intracellular trypsin accumulation and cellular damage responses to supramaximal CCK-8, tobacco toxin, and bile salts in both rodent and human acini. Conclusions: These data demonstrate that EE-LE trafficking acutely controls acinar secretion and the intracellular

Ontario tire recycling and economic development (OnTRED) plan : a market approach to eliminating tire stockpiles and promoting recycled rubber product manufacturing in Ontario

Energy Technology Data Exchange (ETDEWEB)

NONE

2005-05-15

Details of the Ontario Tire Recycling and Economic Development (OnTRED) plan were presented. The plan was developed to address deficiencies in the Ontario Tire Stewardship's (OTS) Scrap Tire Diversion Program plan. The OTS promotes the burning of scrap tires contrary to the Waste Diversion Act, and transfers the financial responsibility for scrap tire management from manufacturers to retailers. The OnTRED plan will attempt to improve the current 87 per cent recovery rate for passenger scrap tires in Ontario, and prevent the formation of any new tire stockpiles. The aim of the OnTRED plan is to ensure reuse and recycling consistent with provisions in the Waste Diversion Act and enhance the existing workings of the tire and scrap tire market through the promotion of reuse and recycling. In addition, the plan aims to minimize administration and compliance costs. A program summary of the OnTRED plan was presented, as well as details of market development plans and buy-recycled rebates. Issues concerning collector registration and transaction tracking were presented, as well as details of brand-owner and first importer pay-in models. Stakeholder roles and responsibilities were reviewed. A budget scenario was presented, as well as a rebate and diversion scenario. It was concluded that Ontario's 87 per cent scrap tire diversion rate can be improved through a focus on patterns of reuse and recycling. 3 tabs., 7 figs.

The N-terminal domains of Vps3 and Vps8 are critical for localization and function of the CORVET tethering complex on endosomes.

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Nadine Epp

Full Text Available Endosomal biogenesis depends on multiple fusion and fission events. For fusion, the heterohexameric CORVET complex as an effector of the endosomal Rab5/Vps21 GTPase has a central function in the initial tethering event. Here, we show that the CORVET-specific Vps3 and Vps8 subunits, which interact with Rab5/Vps21, require their N-terminal domains for localization and function. Surprisingly, CORVET may lack either one of the two N-terminal domains, but not both, to promote protein sorting via the endosome. The dually truncated complex mislocalizes to the cytosol and is impaired in endocytic protein sorting, but not in assembly. Furthermore, the endosomal localization can be rescued by overexpression of Vps21 or one of the truncated CORVET subunits, even though CORVET assembly is not impaired by loss of the N-terminal domains or in strains lacking all endosomal Rab5s and Ypt7. We thus conclude that CORVET requires only its C-terminal domains for assembly and has beyond its putative β-propeller domains additional binding sites for endosomes, which could be important to bind Vps21 and other endosome-specific factors for efficient endosome tethering.

Odin (ANKS1A modulates EGF receptor recycling and stability.

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Jiefei Tong

Full Text Available The ANKS1A gene product, also known as Odin, was first identified as a tyrosine-phosphorylated component of the epidermal growth factor receptor network. Here we show that Odin functions as an effector of EGFR recycling. In EGF-stimulated HEK293 cells tyrosine phosphorylation of Odin was induced prior to EGFR internalization and independent of EGFR-to-ERK signaling. Over-expression of Odin increased EGF-induced EGFR trafficking to recycling endosomes and recycling back to the cell surface, and decreased trafficking to lysosomes and degradation. Conversely, Odin knockdown in both HEK293 and the non-small cell lung carcinoma line RVH6849, which expresses roughly 10-fold more EGF receptors than HEK293, caused decreased EGFR recycling and accelerated trafficking to the lysosome and degradation. By governing the endocytic fate of internalized receptors, Odin may provide a layer of regulation that enables cells to contend with receptor cell densities and ligand concentration gradients that are physiologically and pathologically highly variable.

Novel functions for the endocytic regulatory proteins MICAL-L1 and EHD1 in mitosis.

Science.gov (United States)

Reinecke, James B; Katafiasz, Dawn; Naslavsky, Naava; Caplan, Steve

2015-01-01

During interphase, recycling endosomes mediate the transport of internalized cargo back to the plasma membrane. However, in mitotic cells, recycling endosomes are essential for the completion of cytokinesis, the last phase of mitosis that promotes the physical separation the two daughter cells. Despite recent advances, our understanding of the molecular determinants that regulate recycling endosome dynamics during cytokinesis remains incomplete. We have previously demonstrated that Molecule Interacting with CasL Like-1 (MICAL-L1) and C-terminal Eps15 Homology Domain protein 1 (EHD1) coordinately regulate receptor transport from tubular recycling endosomes during interphase. However, their potential roles in controlling cytokinesis had not been addressed. In this study, we show that MICAL-L1 and EHD1 regulate mitosis. Depletion of either protein resulted in increased numbers of bi-nucleated cells. We provide evidence that bi-nucleation in MICAL-L1- and EHD1-depleted cells is a consequence of impaired recycling endosome transport during late cytokinesis. However, depletion of MICAL-L1, but not EHD1, resulted in aberrant chromosome alignment and lagging chromosomes, suggesting an EHD1-independent function for MICAL-L1 earlier in mitosis. Moreover, we provide evidence that MICAL-L1 and EHD1 differentially influence microtubule dynamics during early and late mitosis. Collectively, our new data suggest several unanticipated roles for MICAL-L1 and EHD1 during the cell cycle. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Intracellular Transport of Vaccinia Virus in HeLa Cells Requires WASH-VPEF/FAM21-Retromer Complexes and Recycling Molecules Rab11 and Rab22

Science.gov (United States)

Hsiao, Jye-Chian; Chu, Li-Wei; Lo, Yung-Tsun; Lee, Sue-Ping; Chen, Tzu-Jung; Huang, Cheng-Yen

2015-01-01

ABSTRACT Vaccinia virus, the prototype of the Orthopoxvirus genus in the family Poxviridae, infects a wide range of cell lines and animals. Vaccinia mature virus particles of the WR strain reportedly enter HeLa cells through fluid-phase endocytosis. However, the intracellular trafficking process of the vaccinia mature virus between cellular uptake and membrane fusion remains unknown. We used live imaging of single virus particles with a combination of various cellular vesicle markers, to track fluorescent vaccinia mature virus particle movement in cells. Furthermore, we performed functional interference assays to perturb distinct vesicle trafficking processes in order to delineate the specific route undertaken by vaccinia mature virus prior to membrane fusion and virus core uncoating in cells. Our results showed that vaccinia virus traffics to early endosomes, where recycling endosome markers Rab11 and Rab22 are recruited to participate in subsequent virus trafficking prior to virus core uncoating in the cytoplasm. Furthermore, we identified WASH-VPEF/FAM21-retromer complexes that mediate endosome fission and sorting of virus-containing vesicles prior to virus core uncoating in the cytoplasm. IMPORTANCE Vaccinia mature virions of the WR strain enter HeLa cells through fluid phase endocytosis. We previously demonstrated that virus-containing vesicles are internalized into phosphatidylinositol 3-phosphate positive macropinosomes, which are then fused with Rab5-positive early endosomes. However, the subsequent process of sorting the virion-containing vesicles prior to membrane fusion remains unclear. We dissected the intracellular trafficking pathway of vaccinia mature virions in cells up to virus core uncoating in cytoplasm. We show that vaccinia mature virions first travel to early endosomes. Subsequent trafficking events require the important endosome-tethered protein VPEF/FAM21, which recruits WASH and retromer protein complexes to the endosome. There, the complex

Extracellular anti-angiogenic proteins augment an endosomal protein trafficking pathway to reach mitochondria and execute apoptosis in HUVECs.

Science.gov (United States)

Chen, Mo; Qiu, Tao; Wu, Jiajie; Yang, Yang; Wright, Graham D; Wu, Min; Ge, Ruowen

2018-03-09

Classic endocytosis destinations include the recycling endosome returning to the plasma membrane or the late endosome (LE) merging with lysosomes for cargo degradation. However, the anti-angiogenic proteins angiostatin and isthmin, are endocytosed and trafficked to mitochondria (Mito) to execute apoptosis of endothelial cells. How these extracellular proteins reach mitochondria remains a mystery. Through confocal and super-resolution fluorescent microscopy, we demonstrate that angiostatin and isthmin are trafficked to mitochondria through the interaction between LE and Mito. Using purified organelles, the LE-Mito interaction is confirmed through in vitro lipid-fusion assay, as well as single vesicle total internal reflection fluorescent microscopy. LE-Mito interaction enables the transfer of not only lipids but also proteins from LE to Mito. Angiostatin and isthmin augment this endosomal protein trafficking pathway and make use of it to reach mitochondria to execute apoptosis. Cell fractionation and biochemical analysis identified that the cytosolic scaffold protein Na+/H+ exchanger regulatory factor 1 (NHERF1) associated with LE and the t-SNARE protein synaptosome-associated protein 25 kDa (SNAP25) associated with Mito form an interaction complex to facilitate LE-Mito interaction. Proximity ligation assay coupled with fluorescent microscopy showed that both NHERF1 and SNAP25 are located at the contacting face between LE and Mito. RNAi knockdown of either NHERF1 or SNAP25 suppressed not only the mitochondrial trafficking of angiostatin and isthmin but also their anti-angiogenic and pro-apoptotic functions. Hence, this study reveals a previously unrealized endosomal protein trafficking pathway from LE to Mito that allows extracellular proteins to reach mitochondria and execute apoptosis.

Recycling at Penn State's Beaver Stadium. "Recycle on the Go" Success Story

Science.gov (United States)

US Environmental Protection Agency, 2009

2009-01-01

With a 13-year-old recycling program, The Pennsylvania State University's (Penn State) Beaver Stadium in the past diverted nearly 30 tons of recyclables per year from local landfills. A new initiative to promote recycling in the stadium's tailgating area has helped Penn State more than triple its old recycling record, collecting 112 tons in 2008.…

PTP1B targets the endosomal sorting machinery

DEFF Research Database (Denmark)

Stuible, Matthew; Abella, Jasmine V; Feldhammer, Matthew

2010-01-01

Dephosphorylation and endocytic down-regulation are distinct processes that together control the signaling output of a variety of receptor tyrosine kinases (RTKs). PTP1B can directly dephosphorylate several RTKs, but it can also promote activation of downstream pathways through largely unknown...... mechanisms. These positive signaling functions likely contribute to the tumor-promoting effect of PTP1B in mouse cancer models. Here, we have identified STAM2, an endosomal protein involved in sorting activated RTKs for lysosomal degradation, as a substrate of PTP1B. PTP1B interacts with STAM2 at defined...... phosphotyrosine sites, and knockdown of PTP1B expression augments STAM2 phosphorylation. Intriguingly, manipulating the expression and phosphorylation state of STAM2 did not have a general effect on epidermal growth factor (EGF)-induced EGF receptor trafficking, degradation, or signaling. Instead, phosphorylated...

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

Cholesterol transfer at endosomal-organelle membrane contact sites.

Science.gov (United States)

Ridgway, Neale D; Zhao, Kexin

2018-06-01

Cholesterol is delivered to the limiting membrane of late endosomes by Niemann-Pick Type C1 and C2 proteins. This review summarizes recent evidence that cholesterol transfer from endosomes to the endoplasmic reticulum and other organelles is mediated by lipid-binding proteins that localize to membrane contact sites (MCS). LDL-cholesterol in the late endosomal/lysosomes is exported to the plasma membrane, where most cholesterol resides, and the endoplasmic reticulum, which harbors the regulatory complexes and enzymes that control the synthesis and esterification of cholesterol. A major advance in dissecting these cholesterol transport pathways was identification of frequent and dynamic MCS between endosomes and the endoplasmic reticulum, peroxisomes and plasma membrane. Positioned at these MCS are members of the oxysterol-binding protein (OSBP) and steroidogenic acute regulatory protein-related lipid-transfer family of lipid transfer proteins that bridge the opposing membranes and directly or indirectly mediate cholesterol transfer. OSBP-related protein 1L (ORP1L), ORP5 and ORP6 mediate cholesterol transfer to the endoplasmic reticulum that regulates cholesterol homeostasis. ORP1L and STARD3 also move cholesterol from the endoplasmic reticulum-to-late endosomal/lysosomes under low-cholesterol conditions to facilitate intraluminal vesicle formation. Cholesterol transport also occurs at MCS with peroxisomes and possibly the plasma membrane. Frequent contacts between organelles and the endo-lysosomal vesicles are sites for bidirectional transfer of cholesterol.

An Essential Role of Hrs/Vps27 in Endosomal Cholesterol Trafficking

Directory of Open Access Journals (Sweden)

Ximing Du

2012-01-01

Full Text Available The endosomal sorting complex required for transport (ESCRT plays a crucial role in the degradation of ubiquitinated endosomal membrane proteins. Here, we report that Hrs, a key protein of the ESCRT-0 complex, is required for the transport of low-density lipoprotein-derived cholesterol from endosomes to the endoplasmic reticulum. This function of Hrs in cholesterol transport is distinct from its previously defined role in lysosomal sorting and downregulation of membrane receptors via the ESCRT pathway. In line with this, knocking down other ESCRT proteins does not cause prominent endosomal cholesterol accumulation. Importantly, the localization and biochemical properties of key cholesterol-sorting proteins, NPC1 and NPC2, appear to be unchanged upon Hrs knockdown. Our data identify Hrs as a regulator of endosomal cholesterol trafficking and provide additional insights into the budding of intralumenal vesicles.

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.

An Inside Job: How Endosomal Na+/H+ Exchangers Link to Autism and Neurological Disease

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Kalyan C. Kondapalli

2014-06-01

Full Text Available Autism imposes a major impediment to childhood development and a huge emotional and financial burden on society. In recent years, there has been rapidly accumulating genetic evidence that links the eNHE, a subset of Na+/H+ exchangers that localize to intracellular vesicles, to a variety of neurological conditions including autism, attention deficit hyperactivity disorder, intellectual disability and epilepsy. By providing a leak pathway for protons pumped by the V-ATPase, eNHE determine luminal pH and regulate cation (Na+, K+ content in early and recycling endosomal compartments. Loss-of-function mutations in eNHE cause hyperacidification of endosomal lumen, as a result of imbalance in pump and leak pathways. Two isoforms, NHE6 and NHE9 are highly expressed in brain, including hippocampus and cortex. Here, we summarize evidence for the importance of luminal cation content and pH on processing, delivery and fate of cargo and on the surface expression and function of membrane receptors and neurotransmitter transporters, drawing upon insights from model organisms and mammalian cells. These studies lead to cellular models of eNHE activity in pre- and post-synaptic neurons and astrocytes, where they could impact synapse development and plasticity. The study of eNHE has provided new insight on the mechanism of autism and other debilitating neurological disorders and opened up new possibilities for therapeutic intervention.

Cell phone recycling experiences in the United States and potential recycling options in Brazil.

Science.gov (United States)

Silveira, Geraldo T R; Chang, Shoou-Yuh

2010-11-01

This paper presents an overview of cell phone recycling programs currently available in the United States. At the same time, it also provides analyses of the current recycling situation and possible recycling alternatives for Brazil. Although there are several recycling options in the United States, collection rates are still only 10% of all potential devices because customers are not aware of these possibilities. The whole system is financially based on reselling refurbished cell phones and recycled materials to developing countries which represent an effective and strong market. Several recyclers offer funds to collection partners who are either charities or who work with charities while obtaining the materials that they need in order to run their operations. A mobile phone recycling system for Brazil considering the United States experience and the Extended Producer Responsibility (EPR) principle is suggested. A deposit/refund/advance-recycling fee is proposed which might be implemented as a voluntary industrial initiative managed by PRO Brazil, a producer responsibility organization. One widespread public-private agreement will integrate all mobile phone stakeholders, and environmental education actions and promotional events will promote citizen's participation. Copyright © 2010 Elsevier Ltd. All rights reserved.

Actin polymerization in the endosomal pathway, but not on the Coxiella-containing vacuole, is essential for pathogen growth.

Directory of Open Access Journals (Sweden)

Heather E Miller

2018-04-01

Full Text Available Coxiella burnetii is an intracellular bacterium that replicates within an expansive phagolysosome-like vacuole. Fusion between the Coxiella-containing vacuole (CCV and late endosomes/multivesicular bodies requires Rab7, the HOPS tethering complex, and SNARE proteins, with actin also speculated to play a role. Here, we investigated the importance of actin in CCV fusion. Filamentous actin patches formed around the CCV membrane that were preferred sites of vesicular fusion. Accordingly, the mediators of endolysosomal fusion Rab7, VAMP7, and syntaxin 8 were concentrated in CCV actin patches. Generation of actin patches required C. burnetii type 4B secretion and host retromer function. Patches decorated with VPS29 and VPS35, components of the retromer, FAM21 and WASH, members of the WASH complex that engage the retromer, and Arp3, a component of the Arp2/3 complex that generates branched actin filaments. Depletion by siRNA of VPS35 or VPS29 reduced CCV actin patches and caused Rab7 to uniformly distribute in the CCV membrane. C. burnetii grew normally in VPS35 or VPS29 depleted cells, as well as WASH-knockout mouse embryo fibroblasts, where CCVs are devoid of actin patches. Endosome recycling to the plasma membrane and trans-Golgi of glucose transporter 1 (GLUT1 and cationic-independent mannose-6-phosphate receptor (CI-M6PR, respectively, was normal in infected cells. However, siRNA knockdown of retromer resulted in aberrant trafficking of GLUT1, but not CI-M6PR, suggesting canonical retrograde trafficking is unaffected by retromer disruption. Treatment with the specific Arp2/3 inhibitor CK-666 strongly inhibited CCV formation, an effect associated with altered endosomal trafficking of transferrin receptor. Collectively, our results show that CCV actin patches generated by retromer, WASH, and Arp2/3 are dispensable for CCV biogenesis and stability. However, Arp2/3-mediated production of actin filaments required for cargo transport within the

Protein complexes and cholesterol in the control of late endosomal dynamicsCholesterol and multi-protein complexes in the control of late endosomal dynamics

NARCIS (Netherlands)

Kant, Rik Henricus Nicolaas van der

2013-01-01

Late endosomal transport is disrupted in several diseases such as Niemann-Pick type C, ARC syndrome and Alzheimer’s disease. This thesis describes the regulation of late endosomal dynamics by cholesterol and multi-protein complexes. We find that cholesterol acts as a cellular tomtom that steers the

Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM)

International Nuclear Information System (INIS)

Duke, Elizabeth M.H.; Razi, Minoo; Weston, Anne; Guttmann, Peter; Werner, Stephan; Henzler, Katja; Schneider, Gerd; Tooze, Sharon A.; Collinson, Lucy M.

2014-01-01

Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10 nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from ‘hotspots’ on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities. - Highlights: • We image whole, unstained mammalian cells using cryo-soft X-ray tomography. • Endosomes are identified using a gold marker for the transferrin receptor. • A new workflow for correlative cryo-fluorescence and cryo-SXT is used to locate early autophagosomes. • Interactions between endosomes, endoplasmic reticulum and forming autophagosomes are mapped in 3D. • Multiple omegasomes are shown to form at ‘hotspots’ on the endoplasmic reticulum

LMTK1 regulates dendritic formation by regulating movement of Rab11A-positive endosomes.

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Takano, Tetsuya; Urushibara, Tomoki; Yoshioka, Nozomu; Saito, Taro; Fukuda, Mitsunori; Tomomura, Mineko; Hisanaga, Shin-Ichi

2014-06-01

Neurons extend two types of neurites-axons and dendrites-that differ in structure and function. Although it is well understood that the cytoskeleton plays a pivotal role in neurite differentiation and extension, the mechanisms by which membrane components are supplied to growing axons or dendrites is largely unknown. We previously reported that the membrane supply to axons is regulated by lemur kinase 1 (LMTK1) through Rab11A-positive endosomes. Here we investigate the role of LMTK1 in dendrite formation. Down-regulation of LMTK1 increases dendrite growth and branching of cerebral cortical neurons in vitro and in vivo. LMTK1 knockout significantly enhances the prevalence, velocity, and run length of anterograde movement of Rab11A-positive endosomes to levels similar to those expressing constitutively active Rab11A-Q70L. Rab11A-positive endosome dynamics also increases in the cell body and growth cone of LMTK1-deficient neurons. Moreover, a nonphosphorylatable LMTK1 mutant (Ser34Ala, a Cdk5 phosphorylation site) dramatically promotes dendrite growth. Thus LMTK1 negatively controls dendritic formation by regulating Rab11A-positive endosomal trafficking in a Cdk5-dependent manner, indicating the Cdk5-LMTK1-Rab11A pathway as a regulatory mechanism of dendrite development as well as axon outgrowth. © 2014 Takano et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Stochastic acidification, activation of hemagglutinin and escape of influenza viruses from an endosome

Science.gov (United States)

Lagache, Thibault; Sieben, Christian; Meyer, Tim; Herrmann, Andreas; Holcman, David

2017-06-01

Influenza viruses enter the cell inside an endosome. During the endosomal journey, acidification triggers a conformational change of the virus spike protein hemagglutinin (HA) that results in escape of the viral genome from the endosome into the cytoplasm. It is still unclear how the interplay between acidification and HA conformation changes affects the kinetics of the viral endosomal escape. We develop here a stochastic model to estimate the change of conformation of HAs inside the endosome nanodomain. Using a Markov process, we model the arrival of protons to HA binding sites and compute the kinetics of their accumulation. We compute the Mean First Passage Time (MFPT) of the number of HA bound sites to a threshold, which is used to estimate the HA activation rate for a given pH concentration. The present analysis reveals that HA proton binding sites possess a high chemical barrier, ensuring a stability of the spike protein at sub-acidic pH. We predict that activating more than 3 adjacent HAs is necessary to trigger endosomal fusion and this configuration prevents premature release of viruses from early endosomes

V-ATPase-dependent luminal acidification is required for endocytic recycling of a yeast cell wall stress sensor, Wsc1p

Energy Technology Data Exchange (ETDEWEB)

Ueno, Kazuma; Saito, Mayu; Nagashima, Makiko; Kojima, Ai; Nishinoaki, Show [Department of Biological Science and Technology, Tokyo University of Science, Niijuku 6-3-1, Katsushika-ku, Tokyo 125-8585 (Japan); Toshima, Junko Y., E-mail: yama_jun@aoni.waseda.jp [Faculty of Science and Engineering, Waseda University, Wakamatsu-cho 2-2, Shinjuku-ku, Tokyo 162-8480 (Japan); Research Center for RNA Science, RIST, Tokyo University of Science, Niijuku 6-3-1, Katsushika-ku, Tokyo 125-8585 (Japan); Toshima, Jiro, E-mail: jtosiscb@rs.noda.tus.ac.jp [Department of Biological Science and Technology, Tokyo University of Science, Niijuku 6-3-1, Katsushika-ku, Tokyo 125-8585 (Japan); Research Center for RNA Science, RIST, Tokyo University of Science, Niijuku 6-3-1, Katsushika-ku, Tokyo 125-8585 (Japan)

2014-01-10

Highlights: •A targeted genome screen identified 5 gene groups affecting Wsc1p recycling. •V-ATPase-dependent luminal acidification is required for Wsc1p recycling. •Activity of V-ATPase might be required for cargo recognition by the retromer complex. -- Abstract: Wsc1p is a major cell wall sensor protein localized at the polarized cell surface. The localization of Wsc1p is maintained by endocytosis and recycling from endosomes back to the cell surface, but changes to the vacuole when cells are subjected to heat stress. Exploiting this unique property of Wsc1p, we screened for yeast single-gene deletion mutants exhibiting defects in Wsc1p trafficking. By expressing 3GFP-tagged Wsc1p in mutants with deleted genes whose function is related to intracellular trafficking, we identified 5 gene groups affecting Wsc1p trafficking, impaired respectively in endocytic internalization, multivesicular body sorting, the GARP complex, endosomal maturation/vacuolar fusion, and V-ATPase. Interestingly, deletion of the VPH1 gene, encoding the V{sub o} subunit of vacuolar-type H{sup +}-ATPase (V-ATPase), led to mis-localization of Wsc1p from the plasma membrane to the vacuole. In addition, disruption of other V-ATPase subunits (vma mutants) also caused defects of Wsc1p trafficking and vacuolar acidification similar to those seen in the vph1Δ mutant. Moreover, we found that deletion of the VPS26 gene, encoding a subunit of the retromer complex, also caused a defect in Wsc1p recycling and mis-localization of Wsc1p to the vacuole. These findings clarified the previously unidentified Wsc1p recycling pathway and requirement of V-ATPase-dependent luminal acidification for Wsc1p recycling.

V-ATPase-dependent luminal acidification is required for endocytic recycling of a yeast cell wall stress sensor, Wsc1p

International Nuclear Information System (INIS)

Ueno, Kazuma; Saito, Mayu; Nagashima, Makiko; Kojima, Ai; Nishinoaki, Show; Toshima, Junko Y.; Toshima, Jiro

2014-01-01

Highlights: •A targeted genome screen identified 5 gene groups affecting Wsc1p recycling. •V-ATPase-dependent luminal acidification is required for Wsc1p recycling. •Activity of V-ATPase might be required for cargo recognition by the retromer complex. -- Abstract: Wsc1p is a major cell wall sensor protein localized at the polarized cell surface. The localization of Wsc1p is maintained by endocytosis and recycling from endosomes back to the cell surface, but changes to the vacuole when cells are subjected to heat stress. Exploiting this unique property of Wsc1p, we screened for yeast single-gene deletion mutants exhibiting defects in Wsc1p trafficking. By expressing 3GFP-tagged Wsc1p in mutants with deleted genes whose function is related to intracellular trafficking, we identified 5 gene groups affecting Wsc1p trafficking, impaired respectively in endocytic internalization, multivesicular body sorting, the GARP complex, endosomal maturation/vacuolar fusion, and V-ATPase. Interestingly, deletion of the VPH1 gene, encoding the V o subunit of vacuolar-type H + -ATPase (V-ATPase), led to mis-localization of Wsc1p from the plasma membrane to the vacuole. In addition, disruption of other V-ATPase subunits (vma mutants) also caused defects of Wsc1p trafficking and vacuolar acidification similar to those seen in the vph1Δ mutant. Moreover, we found that deletion of the VPS26 gene, encoding a subunit of the retromer complex, also caused a defect in Wsc1p recycling and mis-localization of Wsc1p to the vacuole. These findings clarified the previously unidentified Wsc1p recycling pathway and requirement of V-ATPase-dependent luminal acidification for Wsc1p recycling

PIKfyve mediates the motility of late endosomes and lysosomes in neuronal dendrites.

Science.gov (United States)

Tsuruta, Fuminori; Dolmetsch, Ricardo E

2015-09-25

The endosome/lysosome system in the nervous system is critically important for a variety of neuronal functions such as neurite outgrowth, retrograde transport, and synaptic plasticity. In neurons, the endosome/lysosome system is crucial for the activity-dependent internalization of membrane proteins and contributes to the regulation of lipid level on the plasma membrane. Although homeostasis of membrane dynamics plays important roles in the properties of central nervous systems, it has not been elucidated how endosome/lysosome system is regulated. Here, we report that phosphatidylinositol 3-phosphate 5-kinase (PIKfyve) mediates the motility of late endosomes and lysosomes in neuronal dendrites. Endosomes and lysosomes are highly motile in resting neurons, however knockdown of PIKfyve led to a significant reduction in late endosomes and lysosomes motility. We also found that vesicle acidification is crucial for their motility and PIKfyve is associated with this process indirectly. These data suggest that PIKfyve mediates vesicle motility through the regulation of vesicle integrity in neurons. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Mouse polyomavirus enters early endosomes, requires their acidic pH for productive infection, and meets transferrin cargo in rab11-positive endosomes

Czech Academy of Sciences Publication Activity Database

Liebl, D.; Difato, F.; Horníková, L.; Mannová, P.; Štokrová, Jitka; Forstová, J.

2006-01-01

Roč. 80, č. 9 (2006), s. 4610-4622 ISSN 0022-538X R&D Projects: GA ČR(CZ) GA204/03/0593; GA MŠk(CZ) LC545 Institutional research plan: CEZ:AV0Z50520514 Keywords : Polyomavirus internalization and trafficking * Early endosomes * Dependence of infection on endosomal pH Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.341, year: 2006

Recycling of epidermal growth factor-receptor complexes in A431 cells: Identification of dual pathways

International Nuclear Information System (INIS)

Sorkin, A.; Krolenko, S.; Kudrjavtceva, N.; Lazebnik, J.; Teslenko, L.; Soderquist, A.M.; Nikolsky, N.

1991-01-01

The intracellular sorting of EGF-receptor complexes (EGF-RC) has been studied in human epidermoid carcinoma A431 cells. Recycling of EGF was found to occur rapidly after internalization at 37 degrees C. The initial rate of EGF recycling was reduced at 18 degrees C. A significant pool of internalized EGF was incapable of recycling at 18 degrees C but began to recycle when cells were warmed to 37 degrees C. The relative rate of EGF outflow at 37 degrees C from cells exposed to an 18 degrees C temperature block was slower (t1/2 approximately 20 min) than the rate from cells not exposed to a temperature block (t1/2 approximately 5-7 min). These data suggest that there might be both short- and long-time cycles of EGF recycling in A431 cells. Examination of the intracellular EGF-RC dissociation and dynamics of short- and long-time recycling indicated that EGF recycled as EGF-RC. Moreover, EGF receptors that were covalently labeled with a photoactivatable derivative of 125 I-EGF recycled via the long-time pathway at a rate similar to that of 125 I-EGF. Since EGF-RC degradation was also blocked at 18 degrees C, we propose that sorting to the lysosomal and long-time recycling pathway may occur after a highly temperature-sensitive step, presumably in the late endosomes

The ESCRT regulator Did2 maintains the balance between long-distance endosomal transport and endocytic trafficking.

Directory of Open Access Journals (Sweden)

Carl Haag

2017-04-01

Full Text Available In highly polarised cells, like fungal hyphae, early endosomes function in both endocytosis as well as long-distance transport of various cargo including mRNA and protein complexes. However, knowledge on the crosstalk between these seemingly different trafficking processes is scarce. Here, we demonstrate that the ESCRT regulator Did2 coordinates endosomal transport in fungal hyphae of Ustilago maydis. Loss of Did2 results in defective vacuolar targeting, less processive long-distance transport and abnormal shuttling of early endosomes. Importantly, the late endosomal protein Rab7 and vacuolar protease Prc1 exhibit increased shuttling on these aberrant endosomes suggesting defects in endosomal maturation and identity. Consistently, molecular motors fail to attach efficiently explaining the disturbed processive movement. Furthermore, the endosomal mRNP linker protein Upa1 is hardly present on endosomes resulting in defects in long-distance mRNA transport. In conclusion, the ESCRT regulator Did2 coordinates precise maturation of endosomes and thus provides the correct membrane identity for efficient endosomal long-distance transport.

Antigen processing and remodeling of the endosomal pathway: requirements for antigen cross-presentation.

Science.gov (United States)

Compeer, Ewoud Bernardus; Flinsenberg, Thijs Willem Hendrik; van der Grein, Susanna Geertje; Boes, Marianne

2012-01-01

Cross-presentation of endocytosed antigen as peptide/class I major histocompatibility complex complexes plays a central role in the elicitation of CD8(+) T cell clones that mediate anti-viral and anti-tumor immune responses. While it has been clear that there are specific subsets of professional antigen presenting cells capable of antigen cross-presentation, identification of mechanisms involved is still ongoing. Especially amongst dendritic cells (DC), there are specialized subsets that are highly proficient at antigen cross-presentation. We here present a focused survey on the cell biological processes in the endosomal pathway that support antigen cross-presentation. This review highlights DC-intrinsic mechanisms that facilitate the cross-presentation of endocytosed antigen, including receptor-mediated uptake, maturation-induced endosomal sorting of membrane proteins, dynamic remodeling of endosomal structures and cell surface-directed endosomal trafficking. We will conclude with the description of pathogen-induced deviation of endosomal processing, and discuss how immune evasion strategies pertaining endosomal trafficking may preclude antigen cross-presentation.

Erythroid cell mitochondria receive endosomal iron by a "kiss-and-run" mechanism.

Science.gov (United States)

Hamdi, Amel; Roshan, Tariq M; Kahawita, Tanya M; Mason, Anne B; Sheftel, Alex D; Ponka, Prem

2016-12-01

In erythroid cells, more than 90% of transferrin-derived iron enters mitochondria where ferrochelatase inserts Fe 2+ into protoporphyrin IX. However, the path of iron from endosomes to mitochondrial ferrochelatase remains elusive. The prevailing opinion is that, after its export from endosomes, the redox-active metal spreads into the cytosol and mysteriously finds its way into mitochondria through passive diffusion. In contrast, this study supports the hypothesis that the highly efficient transport of iron toward ferrochelatase in erythroid cells requires a direct interaction between transferrin-endosomes and mitochondria (the "kiss-and-run" hypothesis). Using a novel method (flow sub-cytometry), we analyze lysates of reticulocytes after labeling these organelles with different fluorophores. We have identified a double-labeled population definitively representing endosomes interacting with mitochondria, as demonstrated by confocal microscopy. Moreover, we conclude that this endosome-mitochondrion association is reversible, since a "chase" with unlabeled holotransferrin causes a time-dependent decrease in the size of the double-labeled population. Importantly, the dissociation of endosomes from mitochondria does not occur in the absence of holotransferrin. Additionally, mutated recombinant holotransferrin, that cannot release iron, significantly decreases the uptake of 59 Fe by reticulocytes and diminishes 59 Fe incorporation into heme. This suggests that endosomes, which are unable to provide iron to mitochondria, cause a "traffic jam" leading to decreased endocytosis of holotransferrin. Altogether, our results suggest that a molecular mechanism exists to coordinate the iron status of endosomal transferrin with its trafficking. Besides its contribution to the field of iron metabolism, this study provides evidence for a new intracellular trafficking pathway of organelles. Copyright © 2016 Elsevier B.V. All rights reserved.

Amyloid precursor protein and endosomal-lysosomal dysfunction in Alzheimer's disease: inseparable partners in a multifactorial disease.

Science.gov (United States)

Nixon, Ralph A

2017-07-01

Abnormalities of the endosomal-lysosomal network (ELN) are a signature feature of Alzheimer's disease (AD). These include the earliest known cytopathology that is specific to AD and that affects endosomes and induces the progressive failure of lysosomes, each of which are directly linked by distinct mechanisms to neurodegeneration. The origins of ELN dysfunction and β-amyloidogenesis closely overlap, which reflects their common genetic basis, the established early involvement of endosomes and lysosomes in amyloid precursor protein (APP) processing and clearance, and the pathologic effect of certain APP metabolites on ELN functions. Genes that promote β-amyloidogenesis in AD (APP, PSEN1/2, and APOE4) have primary effects on ELN function. The importance of primary ELN dysfunction to pathogenesis is underscored by the mutations in more than 35 ELN-related genes that, thus far, are known to cause familial neurodegenerative diseases even though different pathogenic proteins may be involved. In this article, I discuss growing evidence that implicates AD gene-driven ELN disruptions as not only the antecedent pathobiology that underlies β-amyloidogenesis but also as the essential partner with APP and its metabolites that drive the development of AD, including tauopathy, synaptic dysfunction, and neurodegeneration. The striking amelioration of diverse deficits in animal AD models by remediating ELN dysfunction further supports a need to integrate APP and ELN relationships, including the role of amyloid-β, into a broader conceptual framework of how AD arises, progresses, and may be effectively therapeutically targeted.-Nixon, R. A. Amyloid precursor protein and endosomal-lysosomal dysfunction in Alzheimer's disease: inseparable partners in a multifactorial disease. © FASEB.

deep-orange and carnation define distinct stages in late endosomal biogenesis in Drosophila melanogaster.

Science.gov (United States)

Sriram, V; Krishnan, K S; Mayor, Satyajit

2003-05-12

Endosomal degradation is severely impaired in primary hemocytes from larvae of eye color mutants of Drosophila. Using high resolution imaging and immunofluorescence microscopy in these cells, products of eye color genes, deep-orange (dor) and carnation (car), are localized to large multivesicular Rab7-positive late endosomes containing Golgi-derived enzymes. These structures mature into small sized Dor-negative, Car-positive structures, which subsequently fuse to form tubular lysosomes. Defective endosomal degradation in mutant alleles of dor results from a failure of Golgi-derived vesicles to fuse with morphologically arrested Rab7-positive large sized endosomes, which are, however, normally acidified and mature with wild-type kinetics. This locates the site of Dor function to fusion of Golgi-derived vesicles with the large Rab7-positive endocytic compartments. In contrast, endosomal degradation is not considerably affected in car1 mutant; fusion of Golgi-derived vesicles and maturation of large sized endosomes is normal. However, removal of Dor from small sized Car-positive endosomes is slowed, and subsequent fusion with tubular lysosomes is abolished. Overexpression of Dor in car1 mutant aggravates this defect, implicating Car in the removal of Dor from endosomes. This suggests that, in addition to an independent role in fusion with tubular lysosomes, the Sec1p homologue, Car, regulates Dor function.

Obesity Promotes Alterations in Iron Recycling

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Marta Citelli

2015-01-01

Full Text Available Hepcidin is a key hormone that induces the degradation of ferroportin (FPN, a protein that exports iron from reticuloendothelial macrophages and enterocytes. The aim of the present study was to experimentally evaluate if the obesity induced by a high-fat diet (HFD modifies the expression of FPN in macrophages and enterocytes, thus altering the iron bioavailability. In order to directly examine changes associated with iron metabolism in vivo, C57BL/6J mice were fed either a control or a HFD. Serum leptin levels were evaluated. The hepcidin, divalent metal transporter-1 (DMT1, FPN and ferritin genes were analyzed by real-time polymerase chain reaction. The amount of iron present in both the liver and spleen was determined by flame atomic absorption spectrometry. Ferroportin localization within reticuloendothelial macrophages was observed by immunofluorescence microscopy. Obese animals were found to exhibit increased hepcidin gene expression, while iron accumulated in the spleen and liver. They also exhibited changes in the sublocation of splenic cellular FPN and a reduction in the FPN expression in the liver and the spleen, while no changes were observed in enterocytes. Possible explanations for the increased hepcidin expression observed in HFD animals may include: increased leptin levels, the liver iron accumulation or endoplasmic reticulum (ER stress. Together, the results indicated that obesity promotes changes in iron bioavailability, since it altered the iron recycling function.

Aggregation of endosomal-vacuolar compartments in the Aovps24-deleted strain in the filamentous fungus Aspergillus oryzae

International Nuclear Information System (INIS)

Tatsumi, Akinori; Shoji, Jun-ya; Kikuma, Takashi; Arioka, Manabu; Kitamoto, Katsuhiko

2007-01-01

Previously, we found that deletion of Aovps24, an ortholog of Saccharomyces cerevisiae VPS24, that encodes an ESCRT (endosomal sorting complex required for transport)-III component required for late endosomal function results in fragmented and aggregated vacuoles. Although defective late endosomal function is likely responsible for this phenotype, critical lack of our knowledge on late endosomes in filamentous fungi prevented us from further characterization. In this study, we identified late endosomes of Aspergillus oryzae, by expressing a series of fusion proteins of fluorescent proteins with orthologs of late endosomal proteins. Using these fusion proteins as markers, we observed late endosomes in the wild type strain and the Aovps24 disruptant and demonstrated that late endosomes are aberrantly aggregated in the Aovps24 disruptant. Moreover, we revealed that the aggregated late endosomes have features of vacuoles as well. As deletion of another ESCRT-III component-encoding gene, Aovps2, resulted in similar phenotypes to that in the Aovps24 disruptant, phenotypes of the Aovps24 disruptant are probably due to defective late endosomal function

Structural Basis for Endosomal Targeting by the Bro1 Domain

Science.gov (United States)

Kim, Jaewon; Sitaraman, Sujatha; Hierro, Aitor; Beach, Bridgette M.; Odorizzi, Greg; Hurley, James H.

2010-01-01

Summary Proteins delivered to the lysosome or the yeast vacuole via late endosomes are sorted by the ESCRT complexes and by associated proteins, including Alix and its yeast homolog Bro1. Alix, Bro1, and several other late endosomal proteins share a conserved 160 residue Bro1 domain whose boundaries, structure, and function have not been characterized. The crystal structure of the Bro1 domain of Bro1 reveals a folded core of 367 residues. The extended Bro1 domain is necessary and sufficient for binding to the ESCRT-III subunit Snf7 and for the recruitment of Bro1 to late endosomes. The structure resembles a boomerang with its concave face filled in and contains a triple tetratricopeptide repeat domain as a substructure. Snf7 binds to a conserved hydrophobic patch on Bro1 that is required for protein complex formation and for the protein-sorting function of Bro1. These results define a conserved mechanism whereby Bro1 domain-containing proteins are targeted to endosomes by Snf7 and its orthologs. PMID:15935782

Structure and Function of Vps15 in the Endosomal G Protein Signaling Pathway

Energy Technology Data Exchange (ETDEWEB)

Heenan, Erin J.; Vanhooke, Janeen L.; Temple, Brenda R.; Betts, Laurie; Sondek, John E.; Dohlman, Henrik G.; (UNC)

2009-09-11

G protein-coupled receptors mediate cellular responses to a wide variety of stimuli, including taste, light, and neurotransmitters. In the yeast Saccharomyces cerevisiae, activation of the pheromone pathway triggers events leading to mating. The view had long been held that the G protein-mediated signal occurs principally at the plasma membrane. Recently, it has been shown that the G protein {alpha} subunit Gpa1 can promote signaling at endosomes and requires two components of the sole phosphatidylinositol-3-kinase in yeast, Vps15 and Vps34. Vps15 contains multiple WD repeats and also binds to Gpa1 preferentially in the GDP-bound state; these observations led us to hypothesize that Vps15 may function as a G protein {beta} subunit at the endosome. Here we show an X-ray crystal structure of the Vps15 WD domain that reveals a seven-bladed propeller resembling that of typical G{beta} subunits. We show further that the WD domain is sufficient to bind Gpa1 as well as to Atg14, a potential G{gamma} protein that exists in a complex with Vps15. The Vps15 kinase domain together with the intermediate domain (linking the kinase and WD domains) also contributes to Gpa1 binding and is necessary for Vps15 to sustain G protein signaling. These findings reveal that the Vps15 G{beta}-like domain serves as a scaffold to assemble Gpa1 and Atg14, whereas the kinase and intermediate domains are required for proper signaling at the endosome.

The p25 Subunit of the Dynactin Complex is Required for Dynein-Early Endosome Interaction

Science.gov (United States)

2011-01-01

early endosome movement. In filamentous hyphae , dynein powers the minus end–directed movement of early endosomes (Steinberg and Schuster 2011...observed in time-lapse sequences (Video 1; Abenza et al., 2009). In still images, early endosomes were seen to distribute along the hyphae (Fig. 2 A...nuclear distribution along elongated hyphae and also for the microtubule minus end–directed movement of early endosomes away from the tip (Morris

Nramp1 promotes efficient macrophage recycling of iron following erythrophagocytosis in vivo.

Science.gov (United States)

Soe-Lin, Shan; Apte, Sameer S; Andriopoulos, Billy; Andrews, Marc C; Schranzhofer, Matthias; Kahawita, Tanya; Garcia-Santos, Daniel; Ponka, Prem

2009-04-07

Natural resistance-associated macrophage protein 1 (Nramp1) is a divalent metal transporter expressed exclusively in phagocytic cells. We hypothesized that macrophage Nramp1 may participate in the recycling of iron acquired from phagocytosed senescent erythrocytes. To evaluate the role of Nramp1 in vivo, the iron parameters of WT and KO mice were analyzed after acute and chronic induction of hemolytic anemia. We found that untreated KO mice exhibited greater serum transferrin saturation and splenic iron content with higher duodenal ferroportin (Fpn) and divalent metal transporter 1 (DMT1) expression. Furthermore, hepatocyte iron content and hepcidin mRNA levels were dramatically lower in KO mice, indicating that hepcidin levels can be regulated by low-hepatocyte iron stores despite increased transferrin saturation. After acute treatment with the hemolytic agent phenylhydrazine (Phz), KO mice experienced a significant decrease in transferrin saturation and hematocrit, whereas WT mice were relatively unaffected. After a month-long Phz regimen, KO mice retained markedly increased quantities of iron within the liver and spleen and exhibited more pronounced splenomegaly and reticulocytosis than WT mice. After injection of (59)Fe-labeled heat-damaged reticulocytes, KO animals accumulated erythrophagocytosed (59)Fe within their liver and spleen, whereas WT animals efficiently recycled phagocytosed (59)Fe to the marrow and erythrocytes. These data imply that without Nramp1, iron accumulates within the liver and spleen during erythrophagocytosis and hemolytic anemia, supporting our hypothesis that Nramp1 promotes efficient hemoglobin iron recycling in macrophages. Our observations suggest that mutations in Nramp1 could result in a novel form of human hereditary iron overload.

Characterization of the Mammalian CORVET and HOPS Complexes and Their Modular Restructuring for Endosome Specificity

NARCIS (Netherlands)

van der Kant, Rik; Jonker, Caspar T. H.; Wijdeven, Ruud H.; Bakker, Jeroen; Janssen, Lennert; Klumperman, Judith; Neefjes, Jacques

2015-01-01

Trafficking of cargo through the endosomal system depends on endosomal fusion events mediated by SNARE proteins, Rab-GTPases, and multisubunit tethering complexes. The CORVET and HOPS tethering complexes, respectively, regulate early and late endosomal tethering and have been characterized in detail

α-Taxilin interacts with sorting nexin 4 and participates in the recycling pathway of transferrin receptor.

Directory of Open Access Journals (Sweden)

Hiroshi Sakane

Full Text Available Membrane traffic plays a crucial role in delivering proteins and lipids to their intracellular destinations. We previously identified α-taxilin as a binding partner of the syntaxin family, which is involved in intracellular vesicle traffic. α-Taxilin is overexpressed in tumor tissues and interacts with polymerized tubulin, but the precise function of α-taxilin remains unclear. Receptor proteins on the plasma membrane are internalized, delivered to early endosomes and then either sorted to the lysosome for degradation or recycled back to the plasma membrane. In this study, we found that knockdown of α-taxilin induced the lysosomal degradation of transferrin receptor (TfnR, a well-known receptor which is generally recycled back to the plasma membrane after internalization, and impeded the recycling of transferrin. α-Taxilin was immunoprecipitated with sorting nexin 4 (SNX4, which is involved in the recycling of TfnR. Furthermore, knockdown of α-taxilin decreased the number and length of SNX4-positive tubular structures. We report for the first time that α-taxilin interacts with SNX4 and plays a role in the recycling pathway of TfnR.

Charcot-Marie-Tooth disease-linked protein SIMPLE functions with the ESCRT machinery in endosomal trafficking

OpenAIRE

Lee, Samuel M.; Chin, Lih-Shen; Li, Lian

2012-01-01

Mutations in small integral membrane protein of lysosome/late endosome (SIMPLE) cause autosomal dominant, Charcot-Marie-Tooth disease (CMT) type 1C. The cellular function of SIMPLE is unknown and the pathogenic mechanism of SIMPLE mutations remains elusive. Here, we report that SIMPLE interacted and colocalized with endosomal sorting complex required for transport (ESCRT) components STAM1, Hrs, and TSG101 on early endosomes and functioned with the ESCRT machinery in the control of endosome-to...

Barcoding of GPCR trafficking and signaling through the various trafficking roadmaps by compartmentalized signaling networks.

Science.gov (United States)

Bahouth, Suleiman W; Nooh, Mohammed M

2017-08-01

Proper signaling by G protein coupled receptors (GPCR) is dependent on the specific repertoire of transducing, enzymatic and regulatory kinases and phosphatases that shape its signaling output. Activation and signaling of the GPCR through its cognate G protein is impacted by G protein-coupled receptor kinase (GRK)-imprinted "barcodes" that recruit β-arrestins to regulate subsequent desensitization, biased signaling and endocytosis of the GPCR. The outcome of agonist-internalized GPCR in endosomes is also regulated by sequence motifs or "barcodes" within the GPCR that mediate its recycling to the plasma membrane or retention and eventual degradation as well as its subsequent signaling in endosomes. Given the vast number of diverse sequences in GPCR, several trafficking mechanisms for endosomal GPCR have been described. The majority of recycling GPCR, are sorted out of endosomes in a "sequence-dependent pathway" anchored around a type-1 PDZ-binding module found in their C-tails. For a subset of these GPCR, a second "barcode" imprinted onto specific GPCR serine/threonine residues by compartmentalized kinase networks was required for their efficient recycling through the "sequence-dependent pathway". Mutating the serine/threonine residues involved, produced dramatic effects on GPCR trafficking, indicating that they played a major role in setting the trafficking itinerary of these GPCR. While endosomal SNX27, retromer/WASH complexes and actin were required for efficient sorting and budding of all these GPCR, additional proteins were required for GPCR sorting via the second "barcode". Here we will review recent developments in GPCR trafficking in general and the human β 1 -adrenergic receptor in particular across the various trafficking roadmaps. In addition, we will discuss the role of GPCR trafficking in regulating endosomal GPCR signaling, which promote biochemical and physiological effects that are distinct from those generated by the GPCR signal transduction

A FYVE zinc finger domain protein specifically links mRNA transport to endosome trafficking.

Science.gov (United States)

Pohlmann, Thomas; Baumann, Sebastian; Haag, Carl; Albrecht, Mario; Feldbrügge, Michael

2015-05-18

An emerging theme in cellular logistics is the close connection between mRNA and membrane trafficking. A prominent example is the microtubule-dependent transport of mRNAs and associated ribosomes on endosomes. This coordinated process is crucial for correct septin filamentation and efficient growth of polarised cells, such as fungal hyphae. Despite detailed knowledge on the key RNA-binding protein and the molecular motors involved, it is unclear how mRNAs are connected to membranes during transport. Here, we identify a novel factor containing a FYVE zinc finger domain for interaction with endosomal lipids and a new PAM2-like domain required for interaction with the MLLE domain of the key RNA-binding protein. Consistently, loss of this FYVE domain protein leads to specific defects in mRNA, ribosome, and septin transport without affecting general functions of endosomes or their movement. Hence, this is the first endosomal component specific for mRNP trafficking uncovering a new mechanism to couple mRNPs to endosomes.

Neurotrophin signaling endosomes; biogenesis, regulation, and functions

Science.gov (United States)

Yamashita, Naoya; Kuruvilla, Rejji

2016-01-01

In the nervous system, communication between neurons and their post-synaptic target cells is critical for the formation, refinement and maintenance of functional neuronal connections. Diffusible signals secreted by target tissues, exemplified by the family of neurotrophins, impinge on nerve terminals to influence diverse developmental events including neuronal survival and axonal growth. Key mechanisms of action of target-derived neurotrophins include the cell biological processes of endocytosis and retrograde trafficking of their Trk receptors from growth cones to cell bodies. In this review, we summarize the molecular mechanisms underlying this endosome-mediated signaling, focusing on the instructive role of neurotrophin signaling itself in directing its own trafficking. Recent studies have linked impaired neurotrophin trafficking to neurodevelopmental disorders, highlighting the relevance of neurotrophin endosomes in human health. PMID:27327126

Conformational biosensors reveal GPCR signalling from endosomes

DEFF Research Database (Denmark)

Irannejad, R; Tomshine, Jin C; Tomshine, Jon R

2013-01-01

A long-held tenet of molecular pharmacology is that canonical signal transduction mediated by G-protein-coupled receptor (GPCR) coupling to heterotrimeric G proteins is confined to the plasma membrane. Evidence supporting this traditional view is based on analytical methods that provide limited...... or no subcellular resolution. It has been subsequently proposed that signalling by internalized GPCRs is restricted to G-protein-independent mechanisms such as scaffolding by arrestins, or GPCR activation elicits a discrete form of persistent G protein signalling, or that internalized GPCRs can indeed contribute...... in the early endosome membrane, and that internalized receptors contribute to the overall cellular cyclic AMP response within several minutes after agonist application. These findings provide direct support for the hypothesis that canonical GPCR signalling occurs from endosomes as well as the plasma membrane...

Apolipoprotein E Regulates Amyloid Formation within Endosomes of Pigment Cells

Directory of Open Access Journals (Sweden)

Guillaume van Niel

2015-10-01

Full Text Available Accumulation of toxic amyloid oligomers is a key feature in the pathogenesis of amyloid-related diseases. Formation of mature amyloid fibrils is one defense mechanism to neutralize toxic prefibrillar oligomers. This mechanism is notably influenced by apolipoprotein E variants. Cells that produce mature amyloid fibrils to serve physiological functions must exploit specific mechanisms to avoid potential accumulation of toxic species. Pigment cells have tuned their endosomes to maximize the formation of functional amyloid from the protein PMEL. Here, we show that ApoE is associated with intraluminal vesicles (ILV within endosomes and remain associated with ILVs when they are secreted as exosomes. ApoE functions in the ESCRT-independent sorting mechanism of PMEL onto ILVs and regulates the endosomal formation of PMEL amyloid fibrils in vitro and in vivo. This process secures the physiological formation of amyloid fibrils by exploiting ILVs as amyloid nucleating platforms.

αPIX Is a Trafficking Regulator that Balances Recycling and Degradation of the Epidermal Growth Factor Receptor.

Directory of Open Access Journals (Sweden)

Fanny Kortüm

Full Text Available Endosomal sorting is an essential control mechanism for signaling through the epidermal growth factor receptor (EGFR. We report here that the guanine nucleotide exchange factor αPIX, which modulates the activity of Rho-GTPases, is a potent bimodal regulator of EGFR trafficking. αPIX interacts with the E3 ubiquitin ligase c-Cbl, an enzyme that attaches ubiquitin to EGFR, thereby labelling this tyrosine kinase receptor for lysosomal degradation. We show that EGF stimulation induces αPIX::c-Cbl complex formation. Simultaneously, αPIX and c-Cbl protein levels decrease, which depends on both αPIX binding to c-Cbl and c-Cbl ubiquitin ligase activity. Through interaction αPIX sequesters c-Cbl from EGFR and this results in reduced EGFR ubiquitination and decreased EGFR degradation upon EGF treatment. However, quantitatively more decisive for cellular EGFR distribution than impaired EGFR degradation is a strong stimulating effect of αPIX on EGFR recycling to the cell surface. This function depends on the GIT binding domain of αPIX but not on interaction with c-Cbl or αPIX exchange activity. In summary, our data demonstrate a previously unappreciated function of αPIX as a strong promoter of EGFR recycling. We suggest that the novel recycling regulator αPIX and the degradation factor c-Cbl closely cooperate in the regulation of EGFR trafficking: uncomplexed αPIX and c-Cbl mediate a positive and a negative feedback on EGFR signaling, respectively; αPIX::c-Cbl complex formation, however, results in mutual inhibition, which may reflect a stable condition in the homeostasis of EGF-induced signal flow.

Combustion Byproducts Recycling Consortium

Energy Technology Data Exchange (ETDEWEB)

Paul Ziemkiewicz; Tamara Vandivort; Debra Pflughoeft-Hassett; Y. Paul Chugh; James Hower

2008-08-31

Ashlines: To promote and support the commercially viable and environmentally sound recycling of coal combustion byproducts for productive uses through scientific research, development, and field testing.

EB-promoted recycling of waste tire rubber with polyolefins

Science.gov (United States)

Mészáros, László; Bárány, Tamás; Czvikovszky, Tibor

2012-09-01

Despite the fact that more and more methods and solutions are used in the recycling of polymers, there are still some problems, especially in the recycling of cross-linked materials such as rubber. Usually the biggest problem is the lack of compatibility between the cross-linked rubber and the thermoplastic matrix. In this study we applied ground tire rubber (GTR) as recycled material. The GTR was embedded into polyethylene (PE) and polyethylene/ethylene-vinyl acetate copolymer (PE/EVA) matrices. In order to increase the compatibility of the components electron beam (EB) irradiation was applied. The results showed that the irradiation has a beneficial effect on the polymer-GTR interfacial connection. The EB treatment increased not only the tensile strength but also the elongation at break. The irradiation had also positive effect on the impact strength properties.

Hepatitis C Virus Replication Depends on Endosomal Cholesterol Homeostasis.

Science.gov (United States)

Stoeck, Ina Karen; Lee, Ji-Young; Tabata, Keisuke; Romero-Brey, Inés; Paul, David; Schult, Philipp; Lohmann, Volker; Kaderali, Lars; Bartenschlager, Ralf

2018-01-01

Similar to other positive-strand RNA viruses, hepatitis C virus (HCV) causes massive rearrangements of intracellular membranes, resulting in a membranous web (MW) composed of predominantly double-membrane vesicles (DMVs), the presumed sites of RNA replication. DMVs are enriched for cholesterol, but mechanistic details on the source and recruitment of cholesterol to the viral replication organelle are only partially known. Here we focused on selected lipid transfer proteins implicated in direct lipid transfer at various endoplasmic reticulum (ER)-membrane contact sites. RNA interference (RNAi)-mediated knockdown identified several hitherto unknown HCV dependency factors, such as steroidogenic acute regulatory protein-related lipid transfer domain protein 3 (STARD3), oxysterol-binding protein-related protein 1A and -B (OSBPL1A and -B), and Niemann-Pick-type C1 (NPC1), all residing at late endosome and lysosome membranes and required for efficient HCV RNA replication but not for replication of the closely related dengue virus. Focusing on NPC1, we found that knockdown or pharmacological inhibition caused cholesterol entrapment in lysosomal vesicles concomitant with decreased cholesterol abundance at sites containing the viral replicase factor NS5A. In untreated HCV-infected cells, unesterified cholesterol accumulated at the perinuclear region, partially colocalizing with NS5A at DMVs, arguing for NPC1-mediated endosomal cholesterol transport to the viral replication organelle. Consistent with cholesterol being an important structural component of DMVs, reducing NPC1-dependent endosomal cholesterol transport impaired MW integrity. This suggests that HCV usurps lipid transfer proteins, such as NPC1, at ER-late endosome/lysosome membrane contact sites to recruit cholesterol to the viral replication organelle, where it contributes to MW functionality. IMPORTANCE A key feature of the replication of positive-strand RNA viruses is the rearrangement of the host cell

A FYVE zinc finger domain protein specifically links mRNA transport to endosome trafficking

Science.gov (United States)

Pohlmann, Thomas; Baumann, Sebastian; Haag, Carl; Albrecht, Mario; Feldbrügge, Michael

2015-01-01

An emerging theme in cellular logistics is the close connection between mRNA and membrane trafficking. A prominent example is the microtubule-dependent transport of mRNAs and associated ribosomes on endosomes. This coordinated process is crucial for correct septin filamentation and efficient growth of polarised cells, such as fungal hyphae. Despite detailed knowledge on the key RNA-binding protein and the molecular motors involved, it is unclear how mRNAs are connected to membranes during transport. Here, we identify a novel factor containing a FYVE zinc finger domain for interaction with endosomal lipids and a new PAM2-like domain required for interaction with the MLLE domain of the key RNA-binding protein. Consistently, loss of this FYVE domain protein leads to specific defects in mRNA, ribosome, and septin transport without affecting general functions of endosomes or their movement. Hence, this is the first endosomal component specific for mRNP trafficking uncovering a new mechanism to couple mRNPs to endosomes. DOI: http://dx.doi.org/10.7554/eLife.06041.001 PMID:25985087

Development of the consumption behavior that promotes sustainable society: Focusing on recycling of small waste home appliances

Science.gov (United States)

Ichinose, Takae

2015-04-01

Hiroshima University High School (HUHS) became the first UNESCO Associated School in Japan in 1953, and since then it has practiced ESD in various educational activities in all ranges of education. As a teacher of home economics, I have focused on consumer affairs and encouraged my students to consider what each of them can do as an individual consumer in order to create a sustainable society. In Japan, several acts related to consumer affairs have been enforced in recent years. "Act on Promotion of Consumer Education" was enforced in December 2012, and construction of the "Consumer Citizen Society" was proposed. It places emphasis not only on environmental concerns but also on the initiative of consumers and its influence on social and economic trends. In addition, "Act on Promotion of Recycling of Small Waste Electrical and Electronic Equipment" was enforced in April, 2013. It aims at protecting living environment and healthy development of the national economy by appropriate treatment of waste materials and effective use of resources. For my lessons on "food, clothing and shelter in relation to consumption behavior and environmental problems", I took up "the recycling of small waste home appliances" as the teaching materials to raise awareness on resources recycling. The purpose of the lessons is three-fold: (1) to make students aware of environmental load; (2) to deepen the understanding of the influence which excessive consumption has on developing countries; (3) to encourage the students to think positively toward the solution of the problems. I am currently practicing the lessons, and I have shown below the summary of the instruction. Lesson 1: Give a quiz based on the database on environmental label from Ministry of the Environment website. Then show a film on whereabouts of the hi-tech industrial waste (e-waste). After the film, show some everyday products for which mineral resources are used in order to impress the idea of "urban mine". Lesson 2: Show a

Vps33b pathogenic mutations preferentially affect VIPAS39/SPE-39-positive endosomes.

Science.gov (United States)

Tornieri, Karine; Zlatic, Stephanie A; Mullin, Ariana P; Werner, Erica; Harrison, Robert; L'hernault, Steven W; Faundez, Victor

2013-12-20

Mutations in Vps33 isoforms cause pigment dilution in mice (Vps33a, buff) and Drosophila (car) and the neurogenic arthrogryposis, renal dysfunction and cholestasis syndrome in humans (ARC1, VPS33B). The later disease is also caused by mutations in VIPAS39, (Vps33b interacting protein, apical-basolateral polarity regulator, SPE-39 homolog; ARC2), a protein that interacts with the HOmotypic fusion and Protein Sorting (HOPS) complex, a tether necessary for endosome-lysosome traffic. These syndromes offer insight into fundamental endosome traffic processes unique to metazoans. However, the molecular and cellular mechanisms underlying these mutant phenotypes remain poorly understood. Here we investigate interactions of wild-type and disease-causing mutations in VIPAS39/SPE-39 and Vps33b by yeast two hybrid, immunoprecipitation and quantitative fluorescent microscopy. We find that although few mutations prevent interaction between VIPAS39/SPE-39 and Vps33b, some mutants fragment VIPAS39/SPE-39-positive endosomes, but all mutants alter the subcellular localization of Vps33b to VIPAS39/SPE-39-positive endosomes. Our data suggest that the ARC syndrome may result through impaired VIPAS39/SPE-39 and Vps33b-dependent endosomal maturation or fusion.

The late endosome/lysosome-anchored p18-mTORC1 pathway controls terminal maturation of lysosomes

Energy Technology Data Exchange (ETDEWEB)

Takahashi, Yusuke; Nada, Shigeyuki; Mori, Shunsuke; Soma-Nagae, Taeko; Oneyama, Chitose [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan); Okada, Masato, E-mail: okadam@biken.osaka-u.ac.jp [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan)

2012-01-27

Highlights: Black-Right-Pointing-Pointer p18 is a membrane adaptor that anchors mTORC1 to late endosomes/lysosomes. Black-Right-Pointing-Pointer We examine the role of the p18-mTORC1 pathway in lysosome biogenesis. Black-Right-Pointing-Pointer The loss of p18 causes accumulation of intact late endosomes by arresting lysosome maturation. Black-Right-Pointing-Pointer Inhibition of mTORC1 activity with rapamycin phenocopies the defects of p18 loss. Black-Right-Pointing-Pointer The p18-mTORC1 pathway plays crucial roles in the terminal maturation of lysosomes. -- Abstract: The late endosome/lysosome membrane adaptor p18 (or LAMTOR1) serves as an anchor for the mammalian target of rapamycin complex 1 (mTORC1) and is required for its activation on lysosomes. The loss of p18 causes severe defects in cell growth as well as endosome dynamics, including membrane protein transport and lysosome biogenesis. However, the mechanisms underlying these effects on lysosome biogenesis remain unknown. Here, we show that the p18-mTORC1 pathway is crucial for terminal maturation of lysosomes. The loss of p18 causes aberrant intracellular distribution and abnormal sizes of late endosomes/lysosomes and an accumulation of late endosome specific components, including Rab7, RagC, and LAMP1; this suggests that intact late endosomes accumulate in the absence of p18. These defects are phenocopied by inhibiting mTORC1 activity with rapamycin. Loss of p18 also suppresses the integration of late endosomes and lysosomes, resulting in the defective degradation of tracer proteins. These results suggest that the p18-mTORC1 pathway plays crucial roles in the late stages of lysosomal maturation, potentially in late endosome-lysosome fusion, which is required for processing of various macromolecules.

Endosomal gene expression: a new indicator for prostate cancer patient prognosis?

LENUS (Irish Health Repository)

Johnson, Ian R D

2015-11-10

Prostate cancer continues to be a major cause of morbidity and mortality in men, but a method for accurate prognosis in these patients is yet to be developed. The recent discovery of altered endosomal biogenesis in prostate cancer has identified a fundamental change in the cell biology of this cancer, which holds great promise for the identification of novel biomarkers that can predict disease outcomes. Here we have identified significantly altered expression of endosomal genes in prostate cancer compared to non-malignant tissue in mRNA microarrays and confirmed these findings by qRT-PCR on fresh-frozen tissue. Importantly, we identified endosomal gene expression patterns that were predictive of patient outcomes. Two endosomal tri-gene signatures were identified from a previously published microarray cohort and had a significant capacity to stratify patient outcomes. The expression of APPL1, RAB5A, EEA1, PDCD6IP, NOX4 and SORT1 were altered in malignant patient tissue, when compared to indolent and normal prostate tissue. These findings support the initiation of a case-control study using larger cohorts of prostate tissue, with documented patient outcomes, to determine if different combinations of these new biomarkers can accurately predict disease status and clinical progression in prostate cancer patients.

Human Papillomavirus 16 Infection Induces VAP-Dependent Endosomal Tubulation.

Science.gov (United States)

Siddiqa, Abida; Massimi, Paola; Pim, David; Broniarczyk, Justyna; Banks, Lawrence

2018-03-15

Human papillomavirus (HPV) infection involves complex interactions with the endocytic transport machinery, which ultimately facilitates the entry of the incoming viral genomes into the trans -Golgi network (TGN) and their subsequent nuclear entry during mitosis. The endosomal pathway is a highly dynamic intracellular transport system, which consists of vesicular compartments and tubular extensions, although it is currently unclear whether incoming viruses specifically alter the endocytic machinery. In this study, using MICAL-L1 as a marker for tubulating endosomes, we show that incoming HPV-16 virions induce a profound alteration in global levels of endocytic tubulation. In addition, we also show a critical requirement for the endoplasmic reticulum (ER)-anchored protein VAP in this process. VAP plays an essential role in actin nucleation and endosome-to-Golgi transport. Indeed, the loss of VAP results in a dramatic decrease in the level of endosomal tubulation induced by incoming HPV-16 virions. This is also accompanied by a marked reduction in virus infectivity. In VAP knockdown cells, we see that the defect in virus trafficking occurs after capsid disassembly but prior to localization at the trans -Golgi network, with the incoming virion-transduced DNA accumulating in Vps29/TGN46-positive hybrid vesicles. Taken together, these studies demonstrate that infection with HPV-16 virions induces marked alterations of endocytic transport pathways, some of which are VAP dependent and required for the endosome-to-Golgi transport of the incoming viral L2/DNA complex. IMPORTANCE Human papillomavirus infectious entry involves multiple interactions with the endocytic transport machinery. In this study, we show that incoming HPV-16 virions induce a dramatic increase in endocytic tubulation. This tubulation requires ER-associated VAP, which plays a critical role in ensuring the delivery of cargoes from the endocytic compartments to the trans -Golgi network. Indeed, the loss of

First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro.

Science.gov (United States)

Canfrán-Duque, Alberto; Barrio, Luis C; Lerma, Milagros; de la Peña, Gema; Serna, Jorge; Pastor, Oscar; Lasunción, Miguel A; Busto, Rebeca

2016-03-18

First- and second-generation antipsychotics (FGAs and SGAs, respectively), have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL)-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1'-dioctadecyl-3,3,3,3'-tetramethylindocarbocyanineperchlorate (DiI)-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2) and LBPA (lysobisphosphatidic acid), which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1) and coatomer subunit β (β-COP) were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes' internal milieu induced by haloperidol affects lysosomal functionality.

The N-Myc down regulated Gene1 (NDRG1) Is a Rab4a effector involved in vesicular recycling of E-cadherin.

Science.gov (United States)

Kachhap, Sushant K; Faith, Dennis; Qian, David Z; Shabbeer, Shabana; Galloway, Nathan L; Pili, Roberto; Denmeade, Samuel R; DeMarzo, Angelo M; Carducci, Michael A

2007-09-05

Cell to cell adhesion is mediated by adhesion molecules present on the cell surface. Downregulation of molecules that form the adhesion complex is a characteristic of metastatic cancer cells. Downregulation of the N-myc down regulated gene1 (NDRG1) increases prostate and breast metastasis. The exact function of NDRG1 is not known. Here by using live cell confocal microscopy and in vitro reconstitution, we report that NDRG1 is involved in recycling the adhesion molecule E-cadherin thereby stabilizing it. Evidence is provided that NDRG1 recruits on recycling endosomes in the Trans Golgi network by binding to phosphotidylinositol 4-phosphate and interacts with membrane bound Rab4aGTPase. NDRG1 specifically interacts with constitutively active Rab4aQ67L mutant protein and not with GDP-bound Rab4aS22N mutant proving NDRG1 as a novel Rab4a effector. Transferrin recycling experiments reveals NDRG1 colocalizes with transferrin during the recycling phase. NDRG1 alters the kinetics of transferrin recycling in cells. NDRG1 knockdown cells show a delay in recycling transferrin, conversely NDRG1 overexpressing cells reveal an increase in rate of transferrin recycling. This novel finding of NDRG1 as a recycling protein involved with recycling of E-cadherin will aid in understanding NDRG1 role as a metastasis suppressor protein.

The N-Myc down regulated Gene1 (NDRG1 Is a Rab4a effector involved in vesicular recycling of E-cadherin.

Directory of Open Access Journals (Sweden)

Sushant K Kachhap

2007-09-01

Full Text Available Cell to cell adhesion is mediated by adhesion molecules present on the cell surface. Downregulation of molecules that form the adhesion complex is a characteristic of metastatic cancer cells. Downregulation of the N-myc down regulated gene1 (NDRG1 increases prostate and breast metastasis. The exact function of NDRG1 is not known. Here by using live cell confocal microscopy and in vitro reconstitution, we report that NDRG1 is involved in recycling the adhesion molecule E-cadherin thereby stabilizing it. Evidence is provided that NDRG1 recruits on recycling endosomes in the Trans Golgi network by binding to phosphotidylinositol 4-phosphate and interacts with membrane bound Rab4aGTPase. NDRG1 specifically interacts with constitutively active Rab4aQ67L mutant protein and not with GDP-bound Rab4aS22N mutant proving NDRG1 as a novel Rab4a effector. Transferrin recycling experiments reveals NDRG1 colocalizes with transferrin during the recycling phase. NDRG1 alters the kinetics of transferrin recycling in cells. NDRG1 knockdown cells show a delay in recycling transferrin, conversely NDRG1 overexpressing cells reveal an increase in rate of transferrin recycling. This novel finding of NDRG1 as a recycling protein involved with recycling of E-cadherin will aid in understanding NDRG1 role as a metastasis suppressor protein.

The Vici Syndrome Protein EPG5 Is a Rab7 Effector that Determines the Fusion Specificity of Autophagosomes with Late Endosomes/Lysosomes.

Science.gov (United States)

Wang, Zheng; Miao, Guangyan; Xue, Xue; Guo, Xiangyang; Yuan, Chongzhen; Wang, Zhaoyu; Zhang, Gangming; Chen, Yingyu; Feng, Du; Hu, Junjie; Zhang, Hong

2016-09-01

Mutations in the human autophagy gene EPG5 cause the multisystem disorder Vici syndrome. Here we demonstrated that EPG5 is a Rab7 effector that determines the fusion specificity of autophagosomes with late endosomes/lysosomes. EPG5 is recruited to late endosomes/lysosomes by direct interaction with Rab7 and the late endosomal/lysosomal R-SNARE VAMP7/8. EPG5 also binds to LC3/LGG-1 (mammalian and C. elegans Atg8 homolog, respectively) and to assembled STX17-SNAP29 Qabc SNARE complexes on autophagosomes. EPG5 stabilizes and facilitates the assembly of STX17-SNAP29-VAMP7/8 trans-SNARE complexes, and promotes STX17-SNAP29-VAMP7-mediated fusion of reconstituted proteoliposomes. Loss of EPG5 activity causes abnormal fusion of autophagosomes with various endocytic vesicles, in part due to elevated assembly of STX17-SNAP25-VAMP8 complexes. SNAP25 knockdown partially suppresses the autophagy defect caused by EPG5 depletion. Our study reveals that EPG5 is a Rab7 effector involved in autophagosome maturation, providing insight into the molecular mechanism underlying Vici syndrome. Copyright © 2016 Elsevier Inc. All rights reserved.

A Rab11A/myosin Vb/Rab11-FIP2 complex frames two late recycling steps of langerin from the ERC to the plasma membrane.

Science.gov (United States)

Gidon, Alexandre; Bardin, Sabine; Cinquin, Bertrand; Boulanger, Jerome; Waharte, François; Heliot, Laurent; de la Salle, Henri; Hanau, Daniel; Kervrann, Charles; Goud, Bruno; Salamero, Jean

2012-06-01

A large body of knowledge relating to the constitution of Rab GTPase/Rab effector complexes and their impact on both membrane domain organization and overall membrane trafficking has been built up in recent years. However in the context of the live cell there are still many questions that remain to be answered, such as where and when these complexes assemble and where they perform their primary function(s). We describe here the dynamic processes that take place in the final steps of the Rab11A dependent recycling pathway, in the context of the membrane platform constituted by Myosin Vb, Rab11A, and Rab11-FIP2. We first confirm that a series of previously reported observations obtained during the study of a number of trafficking cargoes also apply to langerin. Langerin is a cargo molecule that traffics through Rab11A-positive membrane domains of the endosomal recycling pathway. In order to explore the relative dynamics of this set of partners, we make extensive use of a combinatory approach of Live-FRET, fast FRAP video, fast confocal and TIRF microscopy modalities. Our data show that the Myosin Vb/Rab11A/Rab11-FIP2 platform is spatially involved in the regulation of langerin trafficking at two distinct sites within live cells, first at the sorting site in the endosomal recycling compartment (ERC) where transport vesicles are formed, and subsequently, in a strict time-defined order, at the very late stage of docking/tethering and fusion of these langerin recycling vesicles to the plasma membrane. © 2012 John Wiley & Sons A/S.

Natural Modulators of Endosomal Toll-Like Receptor-Mediated Psoriatic Skin Inflammation

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Chao-Yang Lai

2017-01-01

Full Text Available Psoriasis is a chronic inflammatory autoimmune disease that can be initiated by excessive activation of endosomal toll-like receptors (TLRs, particularly TLR7, TLR8, and TLR9. Therefore, inhibitors of endosomal TLR activation are being investigated for their ability to treat this disease. The currently approved biological drugs adalimumab, etanercept, infliximab, ustekinumab, ixekizumab, and secukizumab are antibodies against effector cytokines that participate in the initiation and development of psoriasis. Several immune modulatory oligonucleotides and small molecular weight compounds, including IMO-3100, IMO-8400, and CPG-52364, that block the interaction between endosomal TLRs and their ligands are under clinical investigation for their effectiveness in the treatment of psoriasis. In addition, several chemical compounds, including AS-2444697, PF-05387252, PF-05388169, PF-06650833, ML120B, and PHA-408, can inhibit TLR signaling. Although these compounds have demonstrated anti-inflammatory activity in animal models, their therapeutic potential for the treatment of psoriasis has not yet been tested. Recent studies demonstrated that natural compounds derived from plants, fungi, and bacteria, including mustard seed, Antrodia cinnamomea extract, curcumin, resveratrol, thiostrepton, azithromycin, and andrographolide, inhibited psoriasis-like inflammation induced by the TLR7 agonist imiquimod in animal models. These natural modulators employ different mechanisms to inhibit endosomal TLR activation and are administered via different routes. Therefore, they represent candidate psoriasis drugs and might lead to the development of new treatment options.

First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro

Directory of Open Access Journals (Sweden)

Alberto Canfrán-Duque

2016-03-01

Full Text Available First- and second-generation antipsychotics (FGAs and SGAs, respectively, have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1′-dioctadecyl-3,3,3,3′-tetramethylindocarbocyanineperchlorate (DiI-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2 and LBPA (lysobisphosphatidic acid, which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1 and coatomer subunit β (β-COP were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes’ internal milieu induced by haloperidol affects lysosomal functionality.

First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro

Science.gov (United States)

Canfrán-Duque, Alberto; Barrio, Luis C.; Lerma, Milagros; de la Peña, Gema; Serna, Jorge; Pastor, Oscar; Lasunción, Miguel A.; Busto, Rebeca

2016-01-01

First- and second-generation antipsychotics (FGAs and SGAs, respectively), have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL)-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1′-dioctadecyl-3,3,3,3′-tetramethylindocarbocyanineperchlorate (DiI)-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2) and LBPA (lysobisphosphatidic acid), which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1) and coatomer subunit β (β-COP) were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes’ internal milieu induced by haloperidol affects lysosomal functionality. PMID:26999125

The Annexin A1 Receptor FPR2 Regulates the Endosomal Export of Influenza Virus

Directory of Open Access Journals (Sweden)

Fryad Rahman

2018-05-01

Full Text Available The Formyl Peptide Receptor 2 (FPR2 is a novel promising target for the treatment of influenza. During viral infection, FPR2 is activated by annexinA1, which is present in the envelope of influenza viruses; this activation promotes virus replication. Here, we investigated whether blockage of FPR2 would affect the genome trafficking of influenza virus. We found that, upon infection and cell treatment with the specific FPR2 antagonist WRW4 or the anti-FPR2 monoclonal antibody, FN-1D6-AI, influenza viruses were blocked into endosomes. This effect was independent on the strain and was observed for H1N1 and H3N2 viruses. In addition, blocking FPR2signaling in alveolar lung A549 epithelial cells with the monoclonal anti-FPR2 antibody significantly inhibited virus replication. Altogether, these results show that FPR2signaling interferes with the endosomal trafficking of influenza viruses and provides, for the first time, the proof of concept that monoclonal antibodies directed against FPR2 inhibit virus replication. Antibodies-based therapeutics have emerged as attractive reagents in infectious diseases. Thus, this study suggests that the use of anti-FPR2 antibodies against influenza hold great promise for the future.

Promyelocytic leukemia bodies tether to early endosomes during mitosis.

Science.gov (United States)

Palibrk, Vuk; Lång, Emma; Lång, Anna; Schink, Kay Oliver; Rowe, Alexander D; Bøe, Stig Ove

2014-01-01

During mitosis the nuclear envelope breaks down, leading to potential interactions between cytoplasmic and nuclear components. PML bodies are nuclear structures with tumor suppressor and antiviral functions. Early endosomes, on the other hand, are cytoplasmic vesicles involved in transport and growth factor signaling. Here we demonstrate that PML bodies form stable interactions with early endosomes immediately following entry into mitosis. The 2 compartments remain stably associated throughout mitosis and dissociate in the cytoplasm of newly divided daughter cells. We also show that a minor subset of PML bodies becomes anchored to the mitotic spindle poles during cell division. The study demonstrates a stable mitosis-specific interaction between a cytoplasmic and a nuclear compartment.

Rab14 and its exchange factor FAM116 link endocytic recycling and adherens junction stability in migrating cells.

Science.gov (United States)

Linford, Andrea; Yoshimura, Shin-ichiro; Nunes Bastos, Ricardo; Langemeyer, Lars; Gerondopoulos, Andreas; Rigden, Daniel J; Barr, Francis A

2012-05-15

Rab GTPases define the vesicle trafficking pathways underpinning cell polarization and migration. Here, we find that Rab4, Rab11, and Rab14 and the candidate Rab GDP-GTP exchange factors (GEFs) FAM116A and AVL9 are required for cell migration. Rab14 and its GEF FAM116A localize to and act on an intermediate compartment of the transferrin-recycling pathway prior to Rab11 and after Rab5 and Rab4. This Rab14 intermediate recycling compartment has specific functions in migrating cells discrete from early and recycling endosomes. Rab14-depleted cells show increased N-cadherin levels at junctional complexes and cannot resolve cell-cell junctions. This is due to decreased shedding of cell-surface N-cadherin by the ADAM family protease ADAM10/Kuzbanian. In FAM116A- and Rab14-depleted cells, ADAM10 accumulates in a transferrin-positive endocytic compartment, and the cell-surface level of ADAM10 is correspondingly reduced. FAM116 and Rab14 therefore define an endocytic recycling pathway needed for ADAM protease trafficking and regulation of cell-cell junctions. Copyright © 2012 Elsevier Inc. All rights reserved.

The late endosome/lysosome-anchored p18-mTORC1 pathway controls terminal maturation of lysosomes

International Nuclear Information System (INIS)

Takahashi, Yusuke; Nada, Shigeyuki; Mori, Shunsuke; Soma-Nagae, Taeko; Oneyama, Chitose; Okada, Masato

2012-01-01

Highlights: ► p18 is a membrane adaptor that anchors mTORC1 to late endosomes/lysosomes. ► We examine the role of the p18-mTORC1 pathway in lysosome biogenesis. ► The loss of p18 causes accumulation of intact late endosomes by arresting lysosome maturation. ► Inhibition of mTORC1 activity with rapamycin phenocopies the defects of p18 loss. ► The p18-mTORC1 pathway plays crucial roles in the terminal maturation of lysosomes. -- Abstract: The late endosome/lysosome membrane adaptor p18 (or LAMTOR1) serves as an anchor for the mammalian target of rapamycin complex 1 (mTORC1) and is required for its activation on lysosomes. The loss of p18 causes severe defects in cell growth as well as endosome dynamics, including membrane protein transport and lysosome biogenesis. However, the mechanisms underlying these effects on lysosome biogenesis remain unknown. Here, we show that the p18-mTORC1 pathway is crucial for terminal maturation of lysosomes. The loss of p18 causes aberrant intracellular distribution and abnormal sizes of late endosomes/lysosomes and an accumulation of late endosome specific components, including Rab7, RagC, and LAMP1; this suggests that intact late endosomes accumulate in the absence of p18. These defects are phenocopied by inhibiting mTORC1 activity with rapamycin. Loss of p18 also suppresses the integration of late endosomes and lysosomes, resulting in the defective degradation of tracer proteins. These results suggest that the p18-mTORC1 pathway plays crucial roles in the late stages of lysosomal maturation, potentially in late endosome–lysosome fusion, which is required for processing of various macromolecules.

Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery

Science.gov (United States)

Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

2015-06-01

The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds’ escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds’ cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier.

Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery

Science.gov (United States)

Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

2015-01-01

The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds’ escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds’ cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier. PMID:26123532

Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery.

Science.gov (United States)

Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

2015-06-30

The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds' escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds' cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier.

Enhancing endosomal escape of transduced proteins by photochemical internalisation.

Directory of Open Access Journals (Sweden)

Kevin Mellert

Full Text Available Induced internalisation of functional proteins into cultured cells has become an important aspect in a rising number of in vitro and in vivo assays. The endo-lysosomal entrapment of the transduced proteins remains the major problem in all transduction protocols. In this study we compared the efficiency, cytotoxicity and protein targeting of different commercially available transduction reagents by transducing a well-studied fluorescently labelled protein (Atto488-bovine serum albumin into cultured human sarcoma cells. The amount of internalised protein and toxicity differed between the different reagents, but the percentage of transduced cells was consistently high. Furthermore, in all protocols the signals of the transduced Atto488-BSA were predominantly punctual consistent with an endosomal localisation. To overcome the endosomal entrapment, the transduction protocols were combined with a photochemical internalisation (PCI treatment. Using this combination revealed that an endosomal disruption is highly effective in cell penetrating peptide (CPP mediated transduction, whereas lipid-mediated transductions lead to a lower signal spreading throughout the cytosol. No change in the signal distribution could be achieved in treatments using non-lipid polymers as a transduction reagent. Therefore, the combination of protein transduction protocols based on CPPs with the endosomolytic treatment PCI can facilitate protein transduction experiments in vitro.

Enhancing endosomal escape of transduced proteins by photochemical internalisation.

Science.gov (United States)

Mellert, Kevin; Lamla, Markus; Scheffzek, Klaus; Wittig, Rainer; Kaufmann, Dieter

2012-01-01

Induced internalisation of functional proteins into cultured cells has become an important aspect in a rising number of in vitro and in vivo assays. The endo-lysosomal entrapment of the transduced proteins remains the major problem in all transduction protocols. In this study we compared the efficiency, cytotoxicity and protein targeting of different commercially available transduction reagents by transducing a well-studied fluorescently labelled protein (Atto488-bovine serum albumin) into cultured human sarcoma cells. The amount of internalised protein and toxicity differed between the different reagents, but the percentage of transduced cells was consistently high. Furthermore, in all protocols the signals of the transduced Atto488-BSA were predominantly punctual consistent with an endosomal localisation. To overcome the endosomal entrapment, the transduction protocols were combined with a photochemical internalisation (PCI) treatment. Using this combination revealed that an endosomal disruption is highly effective in cell penetrating peptide (CPP) mediated transduction, whereas lipid-mediated transductions lead to a lower signal spreading throughout the cytosol. No change in the signal distribution could be achieved in treatments using non-lipid polymers as a transduction reagent. Therefore, the combination of protein transduction protocols based on CPPs with the endosomolytic treatment PCI can facilitate protein transduction experiments in vitro.

Integration of two RAB5 groups during endosomal transport in plants

Science.gov (United States)

Ebine, Kazuo; Choi, Seung-won; Ichinose, Sakura; Uemura, Tomohiro; Nakano, Akihiko

2018-01-01

RAB5 is a key regulator of endosomal functions in eukaryotic cells. Plants possess two different RAB5 groups, canonical and plant-unique types, which act via unknown counteracting mechanisms. Here, we identified an effector molecule of the plant-unique RAB5 in Arabidopsis thaliana, ARA6, which we designated PLANT-UNIQUE RAB5 EFFECTOR 2 (PUF2). Preferential colocalization with canonical RAB5 on endosomes and genetic interaction analysis indicated that PUF2 coordinates vacuolar transport with canonical RAB5, although PUF2 was identified as an effector of ARA6. Competitive binding of PUF2 with GTP-bound ARA6 and GDP-bound canonical RAB5, together interacting with the shared activating factor VPS9a, showed that ARA6 negatively regulates canonical RAB5-mediated vacuolar transport by titrating PUF2 and VPS9a. These results suggest a unique and unprecedented function for a RAB effector involving the integration of two RAB groups to orchestrate endosomal trafficking in plant cells. PMID:29749929

Characterization of quality recycled gypsum and plasterboard with maximized recycled content

International Nuclear Information System (INIS)

Jiménez-Rivero, J.; García-Navarro, J.

2017-01-01

The quality of secondary materials is imperative to promote a circular economy. In order to improve the way in which the quality of recycled gypsum is assessed, European guidelines on recycled gypsum (RG) quality criteria have been outlined in the framework of the Life+ Gypsum to Gypsum (GtoG) project. Such GtoG guidelines, along with the European Standard on gypsum plasterboard EN 520, provided the basis for this study. During the GtoG project, gypsum recycling and plasterboard manufacturing processes were monitored by testing the gypsum feedstock and the plasterboard produced. The aim of this paper is to discuss the results obtained on relevant parameters that characterize gypsum as a secondary raw material, as well as the resulting product. The minimum requirements were fulfilled by 56% of the RG samples and 86% of the plasterboard with increased RG. [es

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

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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.

Coir dust reinforced recycled polypropylene composites

International Nuclear Information System (INIS)

Santos, Bianca B. dos; Costa, Marysilvia F. da; Thire, Rossana M. da S.M.

2015-01-01

The environmental impacts caused by disposed plastics encourage the search for new alternatives. Recycling polymers leads to the degradation of their mechanical properties, which can be modified by the addition of fillers. In this paper, recycled polypropylene from plastic cups with 2%, 5% and 10% of coir dust were produced with and without the addition of additives. These composites were characterized by tensile tests, X-ray diffraction, differential scanning calorimetry and scanning electron microscopy on the fracture surface. It was verified the effectiveness of the addition of coir dust in improving the elasticity modulus of recycled polypropylene besides the effectiveness of additives used in promoting the adhesion of the powder to the matrix. However, higher levels of coir dust caused the appearance of air bubbles inside the material, which contributed to its embrittlement. The addition of coir dust promoted a decrease in the degree of polypropylene crystallinity. (author)

A Study on the Future Issues Regarding the Small Home Appliance Recycling Law -Based on Comparison with the Other Recycling-Related Laws-

OpenAIRE

小林, 寛

2014-01-01

The Small Home Appliance Recycling Law was enacted in August 2012 and took effect in April 2013 for the purpose of collecting and recycling valuable metals included in used small home appliances such as mobile phones. This Law serves as a promotion scheme, which encourages parties concerned to join the system on a voluntary basis and implements recycling based on the current situation in each region under the cooperation among parties, unlike the Home Appliance Recycling Law enacted in 1998 t...

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

Science.gov (United States)

Lee, Seongju; Chang, Jaerak; Blackstone, Craig

2016-01-01

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

Disturbed vesicular trafficking of membrane proteins in prion disease.

Science.gov (United States)

Uchiyama, Keiji; Miyata, Hironori; Sakaguchi, Suehiro

2013-01-01

The pathogenic mechanism of prion diseases remains unknown. We recently reported that prion infection disturbs post-Golgi trafficking of certain types of membrane proteins to the cell surface, resulting in reduced surface expression of membrane proteins and abrogating the signal from the proteins. The surface expression of the membrane proteins was reduced in the brains of mice inoculated with prions, well before abnormal symptoms became evident. Prions or pathogenic prion proteins were mainly detected in endosomal compartments, being particularly abundant in recycling endosomes. Some newly synthesized membrane proteins are delivered to the surface from the Golgi apparatus through recycling endosomes, and some endocytosed membrane proteins are delivered back to the surface through recycling endosomes. These results suggest that prions might cause neuronal dysfunctions and cell loss by disturbing post-Golgi trafficking of membrane proteins via accumulation in recycling endosomes. Interestingly, it was recently shown that delivery of a calcium channel protein to the cell surface was impaired and its function was abrogated in a mouse model of hereditary prion disease. Taken together, these results suggest that impaired delivery of membrane proteins to the cell surface is a common pathogenic event in acquired and hereditary prion diseases.

Using Behavioural Insights to Promote Food Waste Recycling in Urban Households-Evidence From a Longitudinal Field Experiment.

Science.gov (United States)

Linder, Noah; Lindahl, Therese; Borgström, Sara

2018-01-01

Promoting pro-environmental behaviour amongst urban dwellers is one of today's greatest sustainability challenges. The aim of this study is to test whether an information intervention, designed based on theories from environmental psychology and behavioural economics, can be effective in promoting recycling of food waste in an urban area. To this end we developed and evaluated an information leaflet, mainly guided by insights from nudging and community-based social marketing. The effect of the intervention was estimated through a natural field experiment in Hökarängen, a suburb of Stockholm city, Sweden, and was evaluated using a difference-in-difference analysis. The results indicate a statistically significant increase in food waste recycled compared to a control group in the research area. The data analysed was on the weight of food waste collected from sorting stations in the research area, and the collection period stretched for almost 2 years, allowing us to study the short- and long term effects of the intervention. Although the immediate positive effect of the leaflet seems to have attenuated over time, results show that there was a significant difference between the control and the treatment group, even 8 months after the leaflet was distributed. Insights from this study can be used to guide development of similar pro-environmental behaviour interventions for other urban areas in Sweden and abroad, improving chances of reaching environmental policy goals.

Thermoresponsive pegylated bubble liposome nanovectors for efficient siRNA delivery via endosomal escape

KAUST Repository

Alamoudi, Kholod

2017-05-19

Improving the delivery of siRNA into cancer cells via bubble liposomes. Designing a thermoresponsive pegylated liposome through the introduction of ammonium bicarbonate salt into liposomes so as to control their endosomal escape for gene therapy.A sub-200 nm nanovector was fully characterized and examined for cellular uptake, cytotoxicity, endosomal escape and gene silencing.The siRNA-liposomes were internalized into cancer cells within 5 min and then released siRNAs in the cytosol prior to lysosomal degradation upon external temperature elevation. This was confirmed by confocal bioimaging and gene silencing reaching up to 90% and further demonstrated by the protein inhibition of both target genes.The thermoresponsiveness of ammonium bicarbonate containing liposomes enabled the rapid endosomal escape of the particles and resulted in an efficient gene silencing.

Thermoresponsive pegylated bubble liposome nanovectors for efficient siRNA delivery via endosomal escape

KAUST Repository

Alamoudi, Kholod; Martins, Patricia; Croissant, Jonas G.; Patil, Sachin; Omar, Haneen; Khashab, Niveen M.

2017-01-01

Improving the delivery of siRNA into cancer cells via bubble liposomes. Designing a thermoresponsive pegylated liposome through the introduction of ammonium bicarbonate salt into liposomes so as to control their endosomal escape for gene therapy.A sub-200 nm nanovector was fully characterized and examined for cellular uptake, cytotoxicity, endosomal escape and gene silencing.The siRNA-liposomes were internalized into cancer cells within 5 min and then released siRNAs in the cytosol prior to lysosomal degradation upon external temperature elevation. This was confirmed by confocal bioimaging and gene silencing reaching up to 90% and further demonstrated by the protein inhibition of both target genes.The thermoresponsiveness of ammonium bicarbonate containing liposomes enabled the rapid endosomal escape of the particles and resulted in an efficient gene silencing.

Snapin-regulated late endosomal transport is critical for efficient autophagy-lysosomal function in neurons.

Science.gov (United States)

Cai, Qian; Lu, Li; Tian, Jin-Hua; Zhu, Yi-Bing; Qiao, Haifa; Sheng, Zu-Hang

2010-10-06

Neuron maintenance and survival require late endocytic transport from distal processes to the soma where lysosomes are predominantly localized. Here, we report a role for Snapin in attaching dynein to late endosomes through its intermediate chain (DIC). snapin(-/-) neurons exhibit aberrant accumulation of immature lysosomes, clustering and impaired retrograde transport of late endosomes along processes, reduced lysosomal proteolysis due to impaired delivery of internalized proteins and hydrolase precursors from late endosomes to lysosomes, and impaired clearance of autolysosomes, combined with reduced neuron viability and neurodegeneration. The phenotypes are rescued by expressing the snapin transgene, but not the DIC-binding-defective Snapin-L99K mutant. Snapin overexpression in wild-type neurons enhances late endocytic transport and lysosomal function, whereas expressing the mutant defective in Snapin-DIC coupling shows a dominant-negative effect. Altogether, our study highlights new mechanistic insights into how Snapin-DIC coordinates retrograde transport and late endosomal-lysosomal trafficking critical for autophagy-lysosomal function, and thus neuronal homeostasis. Copyright © 2010 Elsevier Inc. All rights reserved.

EB-promoted recycling of waste tire rubber with polyolefins

International Nuclear Information System (INIS)

Mészáros, László; Bárány, Tamás; Czvikovszky, Tibor

2012-01-01

Despite the fact that more and more methods and solutions are used in the recycling of polymers, there are still some problems, especially in the recycling of cross-linked materials such as rubber. Usually the biggest problem is the lack of compatibility between the cross-linked rubber and the thermoplastic matrix. In this study we applied ground tire rubber (GTR) as recycled material. The GTR was embedded into polyethylene (PE) and polyethylene/ethylene-vinyl acetate copolymer (PE/EVA) matrices. In order to increase the compatibility of the components electron beam (EB) irradiation was applied. The results showed that the irradiation has a beneficial effect on the polymer–GTR interfacial connection. The EB treatment increased not only the tensile strength but also the elongation at break. The irradiation had also positive effect on the impact strength properties. - Highlights: ► In this study ground tire rubber was incorporated into polyethylene (LDPE) matrix. ► Compatibilizing effects of irradiation and ethylene-vinyl acetate were investigated. ► The samples were manufactured by twin-screw extrusion and injection molding. ► Both compatibilizing methods improved the rubber-like properties of the blends. ► This improvement was especially significant when they were applied together.

Using Behavioural Insights to Promote Food Waste Recycling in Urban Households—Evidence From a Longitudinal Field Experiment

Directory of Open Access Journals (Sweden)

Noah Linder

2018-03-01

Full Text Available Promoting pro-environmental behaviour amongst urban dwellers is one of today's greatest sustainability challenges. The aim of this study is to test whether an information intervention, designed based on theories from environmental psychology and behavioural economics, can be effective in promoting recycling of food waste in an urban area. To this end we developed and evaluated an information leaflet, mainly guided by insights from nudging and community-based social marketing. The effect of the intervention was estimated through a natural field experiment in Hökarängen, a suburb of Stockholm city, Sweden, and was evaluated using a difference-in-difference analysis. The results indicate a statistically significant increase in food waste recycled compared to a control group in the research area. The data analysed was on the weight of food waste collected from sorting stations in the research area, and the collection period stretched for almost 2 years, allowing us to study the short- and long term effects of the intervention. Although the immediate positive effect of the leaflet seems to have attenuated over time, results show that there was a significant difference between the control and the treatment group, even 8 months after the leaflet was distributed. Insights from this study can be used to guide development of similar pro-environmental behaviour interventions for other urban areas in Sweden and abroad, improving chances of reaching environmental policy goals.

Using Behavioural Insights to Promote Food Waste Recycling in Urban Households—Evidence From a Longitudinal Field Experiment

Science.gov (United States)

Linder, Noah; Lindahl, Therese; Borgström, Sara

2018-01-01

Promoting pro-environmental behaviour amongst urban dwellers is one of today's greatest sustainability challenges. The aim of this study is to test whether an information intervention, designed based on theories from environmental psychology and behavioural economics, can be effective in promoting recycling of food waste in an urban area. To this end we developed and evaluated an information leaflet, mainly guided by insights from nudging and community-based social marketing. The effect of the intervention was estimated through a natural field experiment in Hökarängen, a suburb of Stockholm city, Sweden, and was evaluated using a difference-in-difference analysis. The results indicate a statistically significant increase in food waste recycled compared to a control group in the research area. The data analysed was on the weight of food waste collected from sorting stations in the research area, and the collection period stretched for almost 2 years, allowing us to study the short- and long term effects of the intervention. Although the immediate positive effect of the leaflet seems to have attenuated over time, results show that there was a significant difference between the control and the treatment group, even 8 months after the leaflet was distributed. Insights from this study can be used to guide development of similar pro-environmental behaviour interventions for other urban areas in Sweden and abroad, improving chances of reaching environmental policy goals. PMID:29623056

Encouraging Vietnamese Household Recycling Behavior: Insights and Implications

Directory of Open Access Journals (Sweden)

The Ninh Nguyen

2017-01-01

Full Text Available This research aims to provide new insights into various determinants affecting household recycling. By focusing on Vietnam, this research also extends knowledge about sustainable behavior in emerging markets, which are the major culprits in terms of greenhouse gas emissions. Hypotheses were developed as a result of the critical review of relevant studies in the fields of marketing, psychology, and economics, and then tested using a quantitative survey data. Structured questionnaires were administered to Vietnamese respondents which yielded 486 usable responses. Multivariate statistics reveal that all the determinants influenced their recycling behavior except for moral norms. Attitude towards the importance of recycling exerted the strongest influence, followed by subjective norms and warm glow respectively. On the other hand, attitude towards the inconvenience of recycling significantly reduced recycling behavior. The research findings have important implications for strategies aimed at promoting recycling behavior. Communication and education programs should emphasize how household recycling contributes to environmental protection, as well as stress intrinsic rewards when recycling. Public media campaigns should feature opinion leaders and attractive communicators, who can effectively apply social pressure to perform recycling behavior. Organizations should also make every effort to make recycling more convenient.

The V-ATPase a2-subunit as a putative endosomal pH-sensor.

Science.gov (United States)

Marshansky, V

2007-11-01

V-ATPase (vesicular H(+)-ATPase)-driven intravesicular acidification is crucial for vesicular trafficking. Defects in vesicular acidification and trafficking have recently been recognized as essential determinants of various human diseases. An important role of endosomal acidification in receptor-ligand dissociation and in activation of lysosomal hydrolytic enzymes is well established. However, the molecular mechanisms by which luminal pH information is transmitted to the cytosolic small GTPases that control trafficking events such as budding, coat formation and fusion are unknown. Here, we discuss our recent discovery that endosomal V-ATPase is a pH-sensor regulating the degradative pathway. According to our model, V-ATPase is responsible for: (i) the generation of a pH gradient between vesicular membranes; (ii) sensing of intravesicular pH; and (iii) transmitting this information to the cytosolic side of the membrane. We also propose the hypothetical molecular mechanism involved in function of the V-ATPase a2-subunit as a putative pH-sensor. Based on extensive experimental evidence on the crucial role of histidine residues in the function of PSPs (pH-sensing proteins) in eukaryotic cells, we hypothesize that pH-sensitive histidine residues within the intra-endosomal loops and/or C-terminal luminal tail of the a2-subunit could also be involved in the pH-sensing function of V-ATPase. However, in order to identify putative pH-sensitive histidine residues and to test this hypothesis, it is absolutely essential that we increase our understanding of the folding and transmembrane topology of the a-subunit isoforms of V-ATPase. Thus the crucial role of intra-endosomal histidine residues in pH-dependent conformational changes of the V-ATPase a2-isoform, its interaction with cytosolic small GTPases and ultimately in its acidification-dependent regulation of the endosomal/lysosomal protein degradative pathway remain to be determined.

Myosin 1g Contributes to CD44 Adhesion Protein and Lipid Rafts Recycling and Controls CD44 Capping and Cell Migration in B Lymphocytes

Directory of Open Access Journals (Sweden)

Orestes López-Ortega

2017-12-01

Full Text Available Cell migration and adhesion are critical for immune system function and involve many proteins, which must be continuously transported and recycled in the cell. Recycling of adhesion molecules requires the participation of several proteins, including actin, tubulin, and GTPases, and of membrane components such as sphingolipids and cholesterol. However, roles of actin motor proteins in adhesion molecule recycling are poorly understood. In this study, we identified myosin 1g as one of the important motor proteins that drives recycling of the adhesion protein CD44 in B lymphocytes. We demonstrate that the lack of Myo1g decreases the cell-surface levels of CD44 and of the lipid raft surrogate GM1. In cells depleted of Myo1g, the recycling of CD44 was delayed, the delay seems to be caused at the level of formation of recycling complex and entry into recycling endosomes. Moreover, a defective lipid raft recycling in Myo1g-deficient cells had an impact both on the capping of CD44 and on cell migration. Both processes required the transportation of lipid rafts to the cell surface to deliver signaling components. Furthermore, the extramembrane was essential for cell expansion and remodeling of the plasma membrane topology. Therefore, Myo1g is important during the recycling of lipid rafts to the membrane and to the accompanied proteins that regulate plasma membrane plasticity. Thus, Myosin 1g contributes to cell adhesion and cell migration through CD44 recycling in B lymphocytes.

Rab11-FIP3 Regulation of Lck Endosomal Traffic Controls TCR Signal Transduction.

Science.gov (United States)

Bouchet, Jérôme; Del Río-Iñiguez, Iratxe; Vázquez-Chávez, Elena; Lasserre, Rémi; Agüera-González, Sonia; Cuche, Céline; McCaffrey, Mary W; Di Bartolo, Vincenzo; Alcover, Andrés

2017-04-01

The role of endosomes in receptor signal transduction is a long-standing question, which remains largely unanswered. The T cell Ag receptor and various components of its proximal signaling machinery are associated with distinct endosomal compartments, but how endosomal traffic affects T cell signaling remains ill-defined. In this article, we demonstrate in human T cells that the subcellular localization and function of the protein tyrosine kinase Lck depends on the Rab11 effector FIP3 (Rab11 family interacting protein-3). FIP3 overexpression or silencing and its ability to interact with Rab11 modify Lck subcellular localization and its delivery to the immunological synapse. Importantly, FIP3-dependent Lck localization controls early TCR signaling events, such as tyrosine phosphorylation of TCRζ, ZAP70, and LAT and intracellular calcium concentration, as well as IL-2 gene expression. Interestingly, FIP3 controls both steady-state and poststimulation phosphotyrosine and calcium levels. Finally, our findings indicate that FIP3 modulates TCR-CD3 cell surface expression via the regulation of steady-state Lck-mediated TCRζ phosphorylation, which in turn controls TCRζ protein levels. This may influence long-term T cell activation in response to TCR-CD3 stimulation. Therefore, our data underscore the importance of finely regulated endosomal traffic in TCR signal transduction and T cell activation leading to IL-2 production. Copyright © 2017 by The American Association of Immunologists, Inc.

Characterization of PEBBLEs as a Tool for Real-Time Measurement of Dictyostelium discoideum Endosomal pH

Directory of Open Access Journals (Sweden)

Everett Moding

2009-01-01

Full Text Available The measurement of intracellular ion concentration change is important for understanding the cellular mechanisms for communication. Recently developed nanosensors, (Photonic Explorers for Biomedical use with Biologically Localized Embedding PEBBLEs, have a number of advantages for measuring ions in cells over established methods using microelectrodes, unbound fluorescent dyes, or NMR. PEBBLE sensors have been shown to work in principle for measuring dynamic ion changes, but few in vivo applications have been demonstrated. We modified the protocol for the fabrication of pH sensing PEBBLEs and developed a protocol for the utilization of these sensors for the monitoring of dynamic pH changes in the endosomes of slime mold Dictyostelium discoideum (D. discoideum. Oregon Green 514-CdSe Quantum Dot PEBBLEs were used to measure real-time pH inside D. discoideum endosomes during cAMP stimulation. Endosomal pH was shown to decrease during cAMP signaling, demonstrating a movement of protons into the endosomes of D. discoideum amoebae.

EB-promoted recycling of waste tire rubber with polyolefins

International Nuclear Information System (INIS)

Meszaros, L.; Barany, T.; Czvikovszky, T.

2011-01-01

Complete text of publication follows. One of the principal problems of common recycling of technical polymers is the thermodynamical incompatibility. The change of the Gibbs 'free energy' during the mixing - determined by the changes of enthalpy and entropy - should be directed towards the miscibility. In most of cases, even between closely related polymers, the compatibility is on low level which leads to phase separation and weak adhesion between the phases. Radiation treatment may help in this problem making the polymers more compatible by creating covalent bonds between the polymer phases. Such problem arises at the recycling of ground tire rubbers (GTR). The rubber properties of GTR could be utilized by mixing the GTR with thermoplastic olefins. With the aid of radiation treatment the rubber-like properties of these types of thermoplastic elastomers can be increased. In this study polyethylene (PE) (50-70 wt%) as thermoplastic olefin, 30 wt% of GTR and ethylene vinyl acetate (EVA) (0-20 wt%) as rubber-like compatibilizing material were used for preparation of a recycled rubber containing blend, processable by thermoplastic processes. The melt-mixing of the samples was made by extrusion and then the test specimens were injection molded. The injection molded samples were than treated by high energy electron beam. On the specimens tensile tests, hysteresis, falling weight impact tests, hardness measurements, and dynamic mechanical thermal analysis (DMA) were carried out. The results proved the benefit of the electron beam treatment.

deep-orange and carnation define distinct stages in late endosomal biogenesis in Drosophila melanogaster

OpenAIRE

Sriram, V.; Krishnan, K.S.; Mayor, Satyajit

2003-01-01

Endosomal degradation is severely impaired in primary hemocytes from larvae of eye color mutants of Drosophila. Using high resolution imaging and immunofluorescence microscopy in these cells, products of eye color genes, deep-orange (dor) and carnation (car), are localized to large multivesicular Rab7-positive late endosomes containing Golgi-derived enzymes. These structures mature into small sized Dor-negative, Car-positive structures, which subsequently fuse to form tubular lysosomes. Defec...

A fluorescence resonance energy transfer-based approach for investigating late endosome-lysosome retrograde fusion events.

Science.gov (United States)

Kaufmann, A M; Goldman, S D B; Krise, J P

2009-03-01

Traditionally, lysosomes have been considered to be a terminal endocytic compartment. Recent studies suggest that lysosomes are quite dynamic, being able to fuse with other late endocytic compartments as well as with the plasma membrane. Here we describe a quantitative fluorescence energy transfer (FRET)-based method for assessing rates of retrograde fusion between terminal lysosomes and late endosomes in living cells. Late endosomes were specifically labeled with 800-nm latex beads that were conjugated with streptavidin and Alexa Fluor 555 (FRET donor). Terminal lysosomes were specifically labeled with 10,000-MW dextran polymers conjugated with biotin and Alexa Fluor 647 (FRET acceptor). Following late endosome-lysosome fusion, the strong binding affinity between streptavidin and biotin brought the donor and acceptor fluorophore molecules into close proximity, thereby facilitating the appearance of a FRET emission signal. Because apparent size restrictions in the endocytic pathway do not permit endocytosed latex beads from reaching terminal lysosomes in an anterograde fashion, the appearance of the FRET signal is consistent with retrograde transport of lysosomal cargo back to late endosomes. We assessed the efficiency of this transport step in fibroblasts affected by different lysosome storage disorders-Niemann-Pick type C, mucolipidosis type IV, and Sandhoff's disease, all of which have a similar lysosomal lipid accumulation phenotype. We report here, for the first time, that these disorders can be distinguished by their rate of transfer of lysosome cargos to late endosomes, and we discuss the implications of these findings for developing new therapeutic strategies.

Small Molecules for Early Endosome-Specific Patch Clamping.

Science.gov (United States)

Chen, Cheng-Chang; Butz, Elisabeth S; Chao, Yu-Kai; Grishchuk, Yulia; Becker, Lars; Heller, Stefan; Slaugenhaupt, Susan A; Biel, Martin; Wahl-Schott, Christian; Grimm, Christian

2017-07-20

To resolve the subcellular distribution of endolysosomal ion channels, we have established a novel experimental approach to selectively patch clamp Rab5 positive early endosomes (EE) versus Rab7/LAMP1-positive late endosomes/lysosomes (LE/LY). To functionally characterize ion channels in endolysosomal membranes with the patch-clamp technique, it is important to develop techniques to selectively enlarge the respective organelles. We found here that two small molecules, wortmannin and latrunculin B, enlarge Rab5-positive EE when combined but not Rab7-, LAMP1-, or Rab11 (RE)-positive vesicles. The two compounds act rapidly, specifically, and are readily applicable in contrast to genetic approaches or previously used compounds such as vacuolin, which enlarges EE, RE, and LE/LY. We apply this approach here to measure currents mediated by TRPML channels, in particular TRPML3, which we found to be functionally active in both EE and LE/LY in overexpressing cells as well as in endogenously expressing CD11b+ lung-tissue macrophages. Copyright © 2017 Elsevier Ltd. All rights reserved.

Phosphatidylinositol 3,5-Bisphosphate-Rich Membrane Domains in Endosomes and Lysosomes.

Science.gov (United States)

Takatori, Sho; Tatematsu, Tsuyako; Cheng, Jinglei; Matsumoto, Jun; Akano, Takuya; Fujimoto, Toyoshi

2016-02-01

Phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2 ) has critical functions in endosomes and lysosomes. We developed a method to define nanoscale distribution of PtdIns(3,5)P2 using freeze-fracture electron microscopy. GST-ATG18-4×FLAG was used to label PtdIns(3,5)P2 and its binding to phosphatidylinositol 3-phosphate (PtdIns(3)P) was blocked by an excess of the p40(phox) PX domain. In yeast exposed to hyperosmotic stress, PtdIns(3,5)P2 was concentrated in intramembrane particle (IMP)-deficient domains in the vacuolar membrane, which made close contact with adjacent membranes. The IMP-deficient domain was also enriched with PtdIns(3)P, but was deficient in Vph1p, a liquid-disordered domain marker. In yeast lacking either PtdIns(3,5)P2 or its effector, Atg18p, the IMP-deficient, PtdIns(3)P-rich membranes were folded tightly to make abnormal tubular structures, thus showing where the vacuolar fragmentation process is arrested when PtdIns(3,5)P2 metabolism is defective. In HeLa cells, PtdIns(3,5)P2 was significantly enriched in the vesicular domain of RAB5- and RAB7-positive endosome/lysosomes of the tubulo-vesicular morphology. This biased distribution of PtdIns(3,5)P2 was also observed using fluorescence microscopy, which further showed enrichment of a retromer component, VPS35, in the tubular domain. This is the first report to show segregation of PtdIns(3,5)P2 -rich and -deficient domains in endosome/lysosomes, which should be important for endosome/lysosome functionality. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

G protein-coupled receptor (GPCR) signaling via heterotrimeric G proteins from endosomes.

Science.gov (United States)

Tsvetanova, Nikoleta G; Irannejad, Roshanak; von Zastrow, Mark

2015-03-13

Some G protein-coupled receptors (GPCRs), in addition to activating heterotrimeric G proteins in the plasma membrane, appear to elicit a "second wave" of G protein activation after ligand-induced internalization. We briefly summarize evidence supporting this view and then discuss what is presently known about the functional significance of GPCR-G protein activation in endosomes. Endosomal activation can shape the cellular response temporally by prolonging its overall duration, and may shape the response spatially by moving the location of intracellular second messenger production relative to effectors. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Cell polarity and patterning by PIN trafficking through early endosomal compartments in Arabidopsis thaliana.

Directory of Open Access Journals (Sweden)

Hirokazu Tanaka

2013-05-01

Full Text Available PIN-FORMED (PIN proteins localize asymmetrically at the plasma membrane and mediate intercellular polar transport of the plant hormone auxin that is crucial for a multitude of developmental processes in plants. PIN localization is under extensive control by environmental or developmental cues, but mechanisms regulating PIN localization are not fully understood. Here we show that early endosomal components ARF GEF BEN1 and newly identified Sec1/Munc18 family protein BEN2 are involved in distinct steps of early endosomal trafficking. BEN1 and BEN2 are collectively required for polar PIN localization, for their dynamic repolarization, and consequently for auxin activity gradient formation and auxin-related developmental processes including embryonic patterning, organogenesis, and vasculature venation patterning. These results show that early endosomal trafficking is crucial for cell polarity and auxin-dependent regulation of plant architecture.

Eps homology domain endosomal transport proteins differentially localize to the neuromuscular junction

Directory of Open Access Journals (Sweden)

Mate Suzanne E

2012-09-01

Full Text Available Abstract Background Recycling of endosomes is important for trafficking and maintenance of proteins at the neuromuscular junction (NMJ. We have previously shown high expression of the endocytic recycling regulator Eps15 homology domain-containing (EHD1 proteinin the Torpedo californica electric organ, a model tissue for investigating a cholinergic synapse. In this study, we investigated the localization of EHD1 and its paralogs EHD2, EHD3, and EHD4 in mouse skeletal muscle, and assessed the morphological changes in EHD1−/− NMJs. Methods Localization of the candidate NMJ protein EHD1 was assessed by confocal microscopy analysis of whole-mount mouse skeletal muscle fibers after direct gene transfer and immunolabeling. The potential function of EHD1 was assessed by specific force measurement and α-bungarotoxin-based endplate morphology mapping in EHD1−/− mouse skeletal muscle. Results Endogenous EHD1 localized to primary synaptic clefts of murine NMJ, and this localization was confirmed by expression of recombinant green fluorescent protein labeled-EHD1 in murine skeletal muscle in vivo. EHD1−/− mouse skeletal muscle had normal histology and NMJ morphology, and normal specific force generation during muscle contraction. The EHD 1–4 proteins showed differential localization in skeletal muscle: EHD2 to muscle vasculature, EHD3 to perisynaptic regions, and EHD4 to perinuclear regions and to primary synaptic clefts, but at lower levels than EHD1. Additionally, specific antibodies raised against mammalian EHD1-4 recognized proteins of the expected mass in the T. californica electric organ. Finally, we found that EHD4 expression was more abundant in EHD1−/− mouse skeletal muscle than in wild-type skeletal muscle. Conclusion EHD1 and EHD4 localize to the primary synaptic clefts of the NMJ. Lack of obvious defects in NMJ structure and muscle function in EHD1−/− muscle may be due to functional compensation by other EHD paralogs.

Blue Box Plus Quinte regional recycling demonstration program

Energy Technology Data Exchange (ETDEWEB)

1992-08-01

The Blue Box Plus recycling program was established in September 1990 in the Quinte region of Ontario. The program was intended to develop the necessary operational information so that the existing program could expand to include mixed plastics, corrugated cardboard, and boxboard. Over 33,000 recycling boxes were distributed over an area covering 15 municipalities with a population base of 95,000. The program showed the willingness of the public to participate in recycling, but advertising and promotion of the program were critical for success. Separation of the recycled materials on the collection trucks was found to be a viable approach and more efficient than sorting at the recycling plant. Adding new materials to be recycled could be done efficiently, and operating costs were in line with those for other programs collecting fewer materials. A cooperative market development with industrial players opened up a new and expanding market for boxboard. 6 figs., 9 tabs.

Lysosomal and endosomal heterogeneity in the liver: A comparison of the intracellular pathways of endocytosis in rat liver cells

International Nuclear Information System (INIS)

Kindberg, G.M.; Tolleshaug, H.; Gjoen, T.; Berg, T.

1991-01-01

Air-filled albumin microspheres, asialoorosomucoid and formaldehyde-treated serum albumin are selectively taken up by endocytosis in rat liver Kupffer cells, parenchymal cells and endothelial cells, respectively. Intracellular transport and degradation of endocytosed material were studied by subcellular fractionation in sucrose and Nycodenz gradients after intravenous injection of the ligand. By using ligands labeled with 125I-tyramine-cellobiose, the subcellular distribution of labeled degradation products can be studied because they are trapped at the site of formation. The results show that the kinetics of intracellular transport are different in hepatic parenchymal, endothelial and Kupffer cells. In endothelial cells, the ligand is associated with two types of endosomes during the first minutes after internalization and then is transferred rapidly to the lysosomes. In parenchymal cells, 125I-tyramine-cellobiose-asialoorosomucoid was located in a relatively slowly sedimenting vesicle during the first minute after internalization and subsequently in denser endosomes. Degradation of 125I-tyramine-cellobiose-asialoorosomucoid in parenchymal cells started later than that of 125I-tyramine-cellobiose-formaldehyde-treated serum albumin in endothelial cells. Furthermore, the ligand seemed to be transferred relatively slowly from endosomes to lysosomes, and most of the undegraded ligand was in the endosomes. The rate-limiting step of proteolysis in parenchymal cells is probably the transport from endosomes to lysosomes. In Kupffer cells, most 125I-tyramine-cellobiose-microspheres are found as undegraded material in very dense endosomes up to 3 hr after injection. After 20 hr, most of the ligand is degraded in lysosomes distributed at a lower density than the endosomes in Nycodenz and sucrose gradients

Gαs regulates Glucagon-Like Peptide 1 Receptor-mediated cyclic AMP generation at Rab5 endosomal compartment

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Shravan Babu Girada

2017-10-01

Conclusions: The findings provide the mechanism of endosomal cyclic AMP generation following GLP-1R activation. We identified the specific compartment that serves as an organizing center to generate endosomal cyclic AMP by internalized activated receptor complex.

Recycle and reduction of waste water in ISL operation

International Nuclear Information System (INIS)

Du Zhiming; Liu Naizhong; Su Xuebin; Li Jianhua; Zou Maoqing; Xing Yongguo

2014-01-01

Sandstone type uranium resources will be promote the main force of natural uranium production in China. The wastewater produced in the process of in-situ leaching mining need to be studied specially, so as to meet the requirements of green mining and realize the recycling of wastewater and decrement. We have researched and adopted including nature groundwater environmental recycling, liquor of precipitation recycling, optimization of elution process, the transformation waste water reduction, water evaporation reduction and a series of technological measures. The field application results show that the wastewater recycling and reduction in the process of production achieved a good environmental protection effect. (authors)

COMMD1 regulates the delta epithelial sodium channel (δENaC) through trafficking and ubiquitination

International Nuclear Information System (INIS)

Chang, Tina; Ke, Ying; Ly, Kevin; McDonald, Fiona J.

2011-01-01

Highlights: → The COMM domain of COMMD1 mediates binding to δENaC. → COMMD1 reduces the cell surface population of δENaC. → COMMD1 increases the population of δENaC-ubiquitin. → Both endogenous and transfected δENaC localize with COMMD1 and transferrin suggesting they are located in early/recycling endosomes. -- Abstract: The delta subunit of the epithelial sodium channel (δENaC) is a member of the ENaC/degenerin family of ion channels. δENaC is distinct from the related α-, β- and γENaC subunits, known for their role in sodium homeostasis and blood pressure control, as δENaC is expressed in brain neurons and activated by external protons. COMMD1 (copper metabolism Murr1 domain 1) was previously found to associate with and downregulate δENaC activity. Here, we show that COMMD1 interacts with δENaC through its COMM domain. Co-expression of δENaC with COMMD1 significantly reduced δENaC surface expression, and led to an increase in δENaC ubiquitination. Immunocytochemical and confocal microscopy studies show that COMMD1 promoted localization of δENaC to the early/recycling endosomal pool where the two proteins were localized together. These results suggest that COMMD1 downregulates δENaC activity by reducing δENaC surface expression through promoting internalization of surface δENaC to an intracellular recycling pool, possibly via enhanced ubiquitination.

COMMD1 regulates the delta epithelial sodium channel ({delta}ENaC) through trafficking and ubiquitination

Energy Technology Data Exchange (ETDEWEB)

Chang, Tina; Ke, Ying; Ly, Kevin [Department of Physiology, University of Otago, P.O. Box 913, Dunedin 9054 (New Zealand); McDonald, Fiona J., E-mail: fiona.mcdonald@otago.ac.nz [Department of Physiology, University of Otago, P.O. Box 913, Dunedin 9054 (New Zealand)

2011-08-05

Highlights: {yields} The COMM domain of COMMD1 mediates binding to {delta}ENaC. {yields} COMMD1 reduces the cell surface population of {delta}ENaC. {yields} COMMD1 increases the population of {delta}ENaC-ubiquitin. {yields} Both endogenous and transfected {delta}ENaC localize with COMMD1 and transferrin suggesting they are located in early/recycling endosomes. -- Abstract: The delta subunit of the epithelial sodium channel ({delta}ENaC) is a member of the ENaC/degenerin family of ion channels. {delta}ENaC is distinct from the related {alpha}-, {beta}- and {gamma}ENaC subunits, known for their role in sodium homeostasis and blood pressure control, as {delta}ENaC is expressed in brain neurons and activated by external protons. COMMD1 (copper metabolism Murr1 domain 1) was previously found to associate with and downregulate {delta}ENaC activity. Here, we show that COMMD1 interacts with {delta}ENaC through its COMM domain. Co-expression of {delta}ENaC with COMMD1 significantly reduced {delta}ENaC surface expression, and led to an increase in {delta}ENaC ubiquitination. Immunocytochemical and confocal microscopy studies show that COMMD1 promoted localization of {delta}ENaC to the early/recycling endosomal pool where the two proteins were localized together. These results suggest that COMMD1 downregulates {delta}ENaC activity by reducing {delta}ENaC surface expression through promoting internalization of surface {delta}ENaC to an intracellular recycling pool, possibly via enhanced ubiquitination.

Current organic waste recycling and the potential for local recycling through urban agriculture in Metro Manila.

Science.gov (United States)

Hara, Yuji; Furutani, Takashi; Murakami, Akinobu; Palijon, Armando M; Yokohari, Makoto

2011-11-01

Using the solid waste management programmes of three barangays (the smallest unit of local government in the Philippines) in Quezon City, Metro Manila, as a case study, this research aimed to further the development of efficient organic waste recycling systems through the promotion of urban agricultural activities on green and vacant spaces. First, the quantity of organic waste and compost produced through ongoing barangay projects was measured. The amount of compost that could potentially be utilized on farmland and vacant land within the barangays was then identified to determine the possibility of a local recycling system. The results indicate that, at present, securing buyers for compost is difficult and, therefore, most compost is distributed to large neighbouring farm villages. However, the present analysis of potential compost use within the barangay demonstrates that a more local compost recycling system is indeed feasible.

Direct binding of retromer to human papillomavirus type 16 minor capsid protein L2 mediates endosome exit during viral infection.

Directory of Open Access Journals (Sweden)

Andreea Popa

2015-02-01

Full Text Available Trafficking of human papillomaviruses to the Golgi apparatus during virus entry requires retromer, an endosomal coat protein complex that mediates the vesicular transport of cellular transmembrane proteins from the endosome to the Golgi apparatus or the plasma membrane. Here we show that the HPV16 L2 minor capsid protein is a retromer cargo, even though L2 is not a transmembrane protein. We show that direct binding of retromer to a conserved sequence in the carboxy-terminus of L2 is required for exit of L2 from the early endosome and delivery to the trans-Golgi network during virus entry. This binding site is different from known retromer binding motifs and can be replaced by a sorting signal from a cellular retromer cargo. Thus, HPV16 is an unconventional particulate retromer cargo, and retromer binding initiates retrograde transport of viral components from the endosome to the trans-Golgi network during virus entry. We propose that the carboxy-terminal segment of L2 protein protrudes through the endosomal membrane and is accessed by retromer in the cytoplasm.

Addressing Criticality in Rare Earth Elements via Permanent Magnets Recycling

Science.gov (United States)

Nlebedim, I. C.; King, A. H.

2018-02-01

Rare earth elements (REEs) are critical for many advanced technologies and are faced with potential supply disruptions. Recycling of permanent magnets (PMs) can be good sources for REEs which can help minimize global dependence on freshly mined REEs, but PMs are rarely recycled. Recycling of PMs has been discussed with respect to improving REEs resource sustainability. Some challenges to be addressed in order to establish industrially deployable technologies for PMs recycling have also been discussed, including profitability, energy efficiency and environmental impacts. Key considerations for promoting circular economy via PMs recycling is proposed with the focus on deciding the target points in the supply chain at which the recycled products will be inserted. Important technical considerations for recycling different forms of waste PMs, including swarfs, slags, shredded and intact hard disk drives magnets, have been presented. The aspects of circular economy considered include reusing magnets, remanufacturing magnets and recovering of REEs from waste PMs.

Radioactive metals disposal and recycling impact modelling

International Nuclear Information System (INIS)

Kemp, N.W.; Lunn, R.J.; Belton, V.; Kockar, I.

2014-01-01

Screening life cycle assessment models developed to investigate hypothetical disposal and recycling options for the Windscale Advanced Gas-cooled Reactor heat exchangers were used to generate more complex models addressing the main UK radioactive metals inventory. Both studies show there are significant environmental advantages in the metals recycling promoted by the current low level waste disposal policies, strategies and plans. Financial benefits from current metals treatment options are supported and offer even greater benefits when applied to the UK radioactive metals inventory as a whole. (authors)

LINGO-1 promotes lysosomal degradation of amyloid-β protein precursor

Directory of Open Access Journals (Sweden)

Rian de Laat

2015-03-01

Full Text Available Sequential proteolytic cleavages of amyloid-β protein precursor (AβPP by β-secretase and γ-secretase generate amyloid β (Aβ peptides, which are thought to contribute to Alzheimer's disease (AD. Much of this processing occurs in endosomes following endocytosis of AβPP from the plasma membrane. However, this pathogenic mode of processing AβPP may occur in competition with lysosomal degradation of AβPP, a common fate of membrane proteins trafficking through the endosomal system. Following up on published reports that LINGO-1 binds and promotes the amyloidogenic processing of AβPP we have examined the consequences of LINGO-1/AβPP interactions. We report that LINGO-1 and its paralogs, LINGO-2 and LINGO-3, decrease processing of AβPP in the amyloidogenic pathway by promoting lysosomal degradation of AβPP. We also report that LINGO-1 levels are reduced in AD brain, representing a possible pathogenic mechanism stimulating the generation of Aβ peptides in AD.

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

An ER-Associated Pathway Defines Endosomal Architecture for Controlled Cargo Transport

NARCIS (Netherlands)

Jongsma, Marlieke L. M.; Berlin, Ilana; Wijdeven, Ruud H. M.; Janssen, Lennert; Janssen, George M. C.; Garstka, Malgorzata A.; Janssen, Hans; Mensink, Mark; van Veelen, Peter A.; Spaapen, Robbert M.; Neefjes, Jacques

2016-01-01

Through a network of progressively maturing vesicles, the endosomal system connects the cell's interior with extracellular space. Intriguingly, this network exhibits a bilateral architecture, comprised of a relatively immobile perinuclear vesicle "cloud" and a highly dynamic peripheral contingent.

The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation*

Science.gov (United States)

Rahbek-Clemmensen, Troels; Bay, Tina; Eriksen, Jacob; Gether, Ulrik; Jørgensen, Trine Nygaard

2014-01-01

The serotonin transporter (SERT) plays a critical role in regulating serotonin signaling by mediating reuptake of serotonin from the extracellular space. The molecular and cellular mechanisms controlling SERT levels in the membrane remain poorly understood. To study trafficking of the surface resident SERT, two functional epitope-tagged variants were generated. Fusion of a FLAG-tagged one-transmembrane segment protein Tac to the SERT N terminus generated a transporter with an extracellular epitope suited for trafficking studies (TacSERT). Likewise, a construct with an extracellular antibody epitope was generated by introducing an HA (hemagglutinin) tag in the extracellular loop 2 of SERT (HA-SERT). By using TacSERT and HA-SERT in antibody-based internalization assays, we show that SERT undergoes constitutive internalization in a dynamin-dependent manner. Confocal images of constitutively internalized SERT demonstrated that SERT primarily co-localized with the late endosomal/lysosomal marker Rab7, whereas little co-localization was observed with the Rab11, a marker of the “long loop” recycling pathway. This sorting pattern was distinct from that of a prototypical recycling membrane protein, the β2-adrenergic receptor. Furthermore, internalized SERT co-localized with the lysosomal marker LysoTracker and not with transferrin. The sorting pattern was further confirmed by visualizing internalization of SERT using the fluorescent cocaine analog JHC1-64 and by reversible and pulse-chase biotinylation assays showing evidence for lysosomal degradation of the internalized transporter. Finally, we found that SERT internalized in response to stimulation with 12-myristate 13-acetate co-localized primarily with Rab7- and LysoTracker-positive compartments. We conclude that SERT is constitutively internalized and that the internalized transporter is sorted mainly to degradation. PMID:24973209

The translocon protein Sec61 mediates antigen transport from endosomes in the cytosol for cross-presentation to CD8(+) T cells.

Science.gov (United States)

Zehner, Matthias; Marschall, Andrea L; Bos, Erik; Schloetel, Jan-Gero; Kreer, Christoph; Fehrenschild, Dagmar; Limmer, Andreas; Ossendorp, Ferry; Lang, Thorsten; Koster, Abraham J; Dübel, Stefan; Burgdorf, Sven

2015-05-19

The molecular mechanisms regulating antigen translocation into the cytosol for cross-presentation are under controversial debate, mainly because direct data is lacking. Here, we have provided direct evidence that the activity of the endoplasmic reticulum (ER) translocon protein Sec61 is essential for endosome-to-cytosol translocation. We generated a Sec61-specific intrabody, a crucial tool that trapped Sec61 in the ER and prevented its recruitment into endosomes without influencing Sec61 activity and antigen presentation in the ER. Expression of this ER intrabody inhibited antigen translocation and cross-presentation, demonstrating that endosomal Sec61 indeed mediates antigen transport across endosomal membranes. Moreover, we showed that the recruitment of Sec61 toward endosomes, and hence antigen translocation and cross-presentation, is dependent on dendritic cell activation by Toll-like receptor (TLR) ligands. These data shed light on a long-lasting question regarding antigen cross-presentation and point out a role of the ER-associated degradation machinery in compartments distinct from the ER. Copyright © 2015 Elsevier Inc. All rights reserved.

Compatibilization of recycled polypropylene and recycled poly (ethylene terephthalate blends with SEBS-g-MA

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Luciana Maria Guadagnini Araujo

2018-03-01

Full Text Available Abstract The compatibilization of recycled PP/PET blend with high and low concentration (20 and 5 phr of elastomer functionalized by maleic anhydride (SEBS-g-MA was achieved. Recycled polypropylene from plastic industry and recycled PET from post-consumer bottles was used. PP/PET blends: 80:20 w/w, 50:50 w/w and 20:80 w/w were prepared in an internal mixer for mechanical properties, thermal properties, morphology and rheological properties. SEBS-g-MA promoted compatibilization of the PP/PET blends and improved their properties. With an increasing compatibilization level, the refinement of morphology was observed in the PET rich blend. Compatibilized blends showed negative deviation in the PET glass transition temperature related to neat PET, demonstrating that compatibilization was very successful. PET crystallization was accelerated in the blends due to PP presence that enhanced nucleation. It was found that the 50/50/20 blend showed huge potential for textile fiber application and that of 80/20/20 showed more intermediary properties than neat polymers.

Shoc2 is targeted to late endosomes and required for Erk1/2 activation in EGF-stimulated cells.

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Emilia Galperin

Full Text Available Shoc2 is the putative scaffold protein that interacts with RAS and RAF, and positively regulates signaling to extracellular signal-regulated protein kinases 1 and 2 (ERK1/2. To elucidate the mechanism by which Shoc2 regulates ERK1/2 activation by the epidermal growth factor (EGF receptor (EGFR, we studied subcellular localization of Shoc2. Upon EGFR activation, endogenous Shoc2 and red fluorescent protein tagged Shoc2 were translocated from the cytosol to a subset of late endosomes containing Rab7. The endosomal recruitment of Shoc2 was blocked by overexpression of a GDP-bound H-RAS (N17S mutant and RNAi knockdown of clathrin, suggesting the requirement of RAS activity and clathrin-dependent endocytosis. RNAi depletion of Shoc2 strongly inhibited activation of ERK1/2 by low, physiological EGF concentrations, which was rescued by expression of wild-type recombinant Shoc2. In contrast, the Shoc2 (S2G mutant, that is myristoylated and found in patients with the Noonan-like syndrome, did not rescue ERK1/2 activation in Shoc2-depleted cells. Shoc2 (S2G was not located in late endosomes but was present on the plasma membrane and early endosomes. These data suggest that targeting of Shoc2 to late endosomes may facilitate EGFR-induced ERK activation under physiological conditions of cell stimulation by EGF, and therefore, may be involved in the spatiotemporal regulation of signaling through the RAS-RAF module.

Hanford recycling

Energy Technology Data Exchange (ETDEWEB)

Leonard, I.M.

1996-09-01

This paper is a study of the past and present recycling efforts on the Hanford site and options for future improvements in the recycling program. Until 1996, recycling goals were voluntarily set by the waste generators: this year, DOE has imposed goals for all its sites to accomplish by 1999. Hanford is presently meeting the voluntary site goals, but may not be able to meet all the new DOE goals without changes to the program. Most of these new DOE goals are recycling goals: * Reduce the generation of radioactive (low-level) waste from routine operations 50 percent through source reduction and recycling. * Reduce the generation of low-level mixed waste from routine operations 50 percent through source reduction and recycling. * Reduce the generation of hazardous waste from routine operations 50 percent through source reduction and recycling. * Recycle 33 percent of the sanitary waste from all operations. * Increase affirmative procurement of EPA-designated recycled items to 100 percent. The Hanford recycling program has made great strides-there has been a 98 percent increase in the amount of paper recycled since its inception in 1990. Hanford recycles paper, chemicals cardboard, tires, oil, batteries, rags, lead weights, fluorescent tubes, aerosol products, concrete, office furniture, computer software, drums, toner cartridges, and scrap metal. Many other items are recycled or reused by individual groups on a one time basis without a formal contract. Several contracts are closed-loop contracts which involve all parts of the recycle loop. Considerable savings are generated from recycling, and much more is possible with increased attention and improvements to this program. General methods for improving the recycling program to ensure that the new goals can be met are: a Contract and financial changes 0 Tracking database and methods improvements 0 Expanded recycling efforts. Specifically, the Hanford recycling program would be improved by: 0 Establishing one overall

Recycled sand in lime-based mortars.

Science.gov (United States)

Stefanidou, M; Anastasiou, E; Georgiadis Filikas, K

2014-12-01

The increasing awareness of the society about safe guarding heritage buildings and at the same time protecting the environment promotes strategies of combining principles of restoration with environmentally friendly materials and techniques. Along these lines, an experimental program was carried out in order to investigate the possibility of producing repair, lime-based mortars used in historic buildings incorporating secondary materials. The alternative material tested was recycled fine aggregates originating from mixed construction and demolition waste. Extensive tests on the raw materials have been performed and mortar mixtures were produced using different binding systems with natural, standard and recycled sand in order to compare their mechanical, physical and microstructure properties. The study reveals the improved behavior of lime mortars, even at early ages, due to the reaction of lime with the Al and Si constituents of the fine recycled sand. The role of the recycled sand was more beneficial in lime mortars rather than the lime-pozzolan or lime-pozzolan-cement mortars as a decrease in their performance was recorded in the latter cases due to the mortars' structure. Copyright © 2014 Elsevier Ltd. All rights reserved.

Augmenting the Efficacy of Immunotoxins and Other Targeted Protein Toxins by Endosomal Escape Enhancers

Directory of Open Access Journals (Sweden)

Hendrik Fuchs

2016-07-01

Full Text Available The toxic moiety of almost all protein-based targeted toxins must enter the cytosol of the target cell to mediate its fatal effect. Although more than 500 targeted toxins have been investigated in the past decades, no antibody-targeted protein toxin has been approved for tumor therapeutic applications by the authorities to date. Missing efficacy can be attributed in many cases to insufficient endosomal escape and therefore subsequent lysosomal degradation of the endocytosed toxins. To overcome this drawback, many strategies have been described to weaken the membrane integrity of endosomes. This comprises the use of lysosomotropic amines, carboxylic ionophores, calcium channel antagonists, various cell-penetrating peptides of viral, bacterial, plant, animal, human and synthetic origin, other organic molecules and light-induced techniques. Although the efficacy of the targeted toxins was typically augmented in cell culture hundred or thousand fold, in exceptional cases more than million fold, the combination of several substances harbors new problems including additional side effects, loss of target specificity, difficulties to determine the therapeutic window and cell type-dependent variations. This review critically scrutinizes the chances and challenges of endosomal escape enhancers and their potential role in future developments.

Self-care in individuals who recycle garbage (recyclers of Medellín, Colombia, 2005

Directory of Open Access Journals (Sweden)

Yolanda L. López A

2009-08-01

Full Text Available Objective: to understand the conceptions, attitudes and practices about self-care among informal recyclers in Medellín. Methodology: a qualitative study from the perspective of recyclers not belonging to any guild in Medellín in 2005 was developed. A total amount of 52 interviews with previous informed consent were applied, plus non-participant observation and field diary. Results: self-care is understood as the constant protection of the body in order to avoid the occurrence of diseases. It includes aspects of hygiene, diet, medical consultation, proper use of leisure time and utilization of instruments for personal protection, such as gloves, face masks, caps, and ear plugs among other devices. Discussion: disease prevention goes beyond biological aspects and becomes part of a social dimension for people in which the most important features are the consequences for themselves, their labor team and their families. Recyclers have knowledge on prevention and practices that promote health and well-being, but the latter are not applied because they diminish their job performance, or they do not consider them to be necessary, do not feel like executing them nor they lack the economic resources to assume them. Autocare practices of recyclers are mainly supported by the primacy of the team history than by the scientific knowledge concerning it.

Proteolytic processing of epidermal growth factor within endosomes

International Nuclear Information System (INIS)

Gorman, R.M.; Savage, C.R. Jr.; Poretz, R.D.; Schaudies, R.P.

1986-01-01

The authors have reported previously that EGF enters 3 biochemically distinct non-lysosomal intracellular compartments prior to detection within lysosomes. Earlier studies have demonstrated that EGF is processes by sequential removal of 1, 4 and 1 aminoacyl residues at the C-terminus. The final form, which lacks the 6 residues, accumulates in secondary lysosomes. After subcellular fractionation of fibroblasts exposed to 125 I-EGF, ligand is detected with 3 non-lysosomal endocytic compartments and is fully processed prior to entrance into secondary lysosome. Following internalization, EGF enters an early endosomal compartment (E 1 ). The composition of the ligand (60%, -1 form; 40%, native form) represents an enhancement of the -1 form relative to that on the plasma membrane following the 90 min, 0 0 binding period. The proportion of different EGF forms in E 1 remains constant through the 2 min pulse and chase periods up to 30 min. However, in the ultimate endosomal compartment, E 4 , the proportion of the -6 form increases from 25% at 15 min to greater than 75% in 30 min, with a concomitant decrease of the -1 and -5 forms. Secondary lysosomes contain an EGF composition similar to that found in E 4 at 30 min. Accordingly, it appears that EGF is processed to the -6 form following passage through E 1 and during its tenure in E 4

The Processed Amino-Terminal Fragment of Human TLR7 Acts as a Chaperone To Direct Human TLR7 into Endosomes

Science.gov (United States)

Shepherd, Dawn; Booth, Sarah; Waithe, Dominic; Reis e Sousa, Caetano

2015-01-01

TLR7 mediates innate immune responses to viral RNA in endocytic compartments. Mouse and human (h)TLR7 undergo proteolytic cleavage, resulting in the generation of a C-terminal fragment that accumulates in endosomes and associates with the signaling adaptor MyD88 upon receptor triggering by TLR7 agonists. Although mouse TLR7 is cleaved in endosomes by acidic proteases, hTLR7 processing can occur at neutral pH throughout the secretory pathway through the activity of furin-like proprotein convertases. However, the mechanisms by which cleaved hTLR7 reaches the endosomal compartment remain unclear. In this study, we demonstrate that, after hTLR7 proteolytic processing, the liberated amino (N)-terminal fragment remains bound to the C terminus through disulfide bonds and provides key trafficking information that ensures correct delivery of the complex to endosomal compartments. In the absence of the N-terminal fragment, the C-terminal fragment is redirected to the cell surface, where it is functionally inactive. Our data reveal a novel role for the N terminus of hTLR7 as a molecular chaperone that provides processed hTLR7 with the correct targeting instructions to reach the endosomal compartment, hence ensuring its biological activity and preventing inadvertent cell surface responses to self-RNA. PMID:25917086

RCP-driven α5β1 recycling suppresses Rac and promotes RhoA activity via the RacGAP1-IQGAP1 complex.

Science.gov (United States)

Jacquemet, Guillaume; Green, David M; Bridgewater, Rebecca E; von Kriegsheim, Alexander; Humphries, Martin J; Norman, Jim C; Caswell, Patrick T

2013-09-16

Inhibition of αvβ3 or expression of mutant p53 promotes invasion into fibronectin (FN)-containing extracellular matrix (ECM) by enhancing Rab-coupling protein (RCP)-dependent recycling of α5β1 integrin. RCP and α5β1 cooperatively recruit receptor tyrosine kinases, including EGFR1, to regulate their trafficking and downstream signaling via protein kinase B (PKB)/Akt, which, in turn, promotes invasive migration. In this paper, we identify a novel PKB/Akt substrate, RacGAP1, which is phosphorylated as a consequence of RCP-dependent α5β1 trafficking. Phosphorylation of RacGAP1 promotes its recruitment to IQGAP1 at the tips of invasive pseudopods, and RacGAP1 then locally suppresses the activity of the cytoskeletal regulator Rac and promotes the activity of RhoA in this subcellular region. This Rac to RhoA switch promotes the extension of pseudopodial processes and invasive migration into FN-containing matrices, in a RhoA-dependent manner. Thus, the localized endocytic trafficking of α5β1 within the tips of invasive pseudopods elicits signals that promote the reorganization of the actin cytoskeleton, protrusion, and invasion into FN-rich ECM.

Decoupling Internalization, Acidification and Phagosmal-Endosomal/Iysosomal Phagocytosis of Internalin A coated Beads in epithelial cells

Energy Technology Data Exchange (ETDEWEB)

Blanchette, C D; Woo, Y; Thomas, C; Shen, N; Sulchek, T A; Hiddessen, A L

2008-12-22

Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established, and in several cases, it was treated as a one-step process. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells, such as epithelial cells. Therefore, in this study, we developed a simple and novel method to decouple and accurately measure particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK) and Caco-2 epithelial cells. Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA), a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated internalization. We achieved independent measurements of the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads) with antibody quenching, pH sensitive dyes and endosomal/lysosomal dyes, as follows: the rate of InlA bead internalization was measured via antibody quenching of a pH independent dye (Alexa488) conjugated to InlA-beads, the rate at which phagosomes containing internalized InlA beads became acidified was measured using a pH dependent dye (FITC) conjugated to the beads and the rate of phagosomal-endosomal/lysosomal fusion was measured using a combination of unlabeled InlA-beads and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we also exploited the phagosomal acidification

HIV-1 Nef binds with human GCC185 protein and regulates mannose 6 phosphate receptor recycling

Energy Technology Data Exchange (ETDEWEB)

Kumar, Manjeet; Kaur, Supinder; Nazir, Aamir; Tripathi, Raj Kamal, E-mail: rajkamalcdri@gmail.com

2016-05-20

HIV-1 Nef modulates cellular function that enhances viral replication in vivo which culminate into AIDS pathogenesis. With no enzymatic activity, Nef regulates cellular function through host protein interaction. Interestingly, trans-cellular introduction of recombinant Nef protein in Caenorhabditis elegans results in AIDS like pathogenesis which might share common pathophysiology because the gene sequence of C. elegans and humans share considerable homology. Therefore employing C. elegans based initial screen complemented with sequence based homology search we identified GCC185 as novel host protein interacting with HIV-1 Nef. The detailed molecular characterization revealed N-terminal EEEE{sub 65} acidic domain of Nef as key region for interaction. GCC185 is a tethering protein that binds with Rab9 transport vesicles. Our results show that Nef-GCC185 interaction disrupts Rab9 interaction resulting in delocalization of CI-MPR (cation independent Mannose 6 phosphate receptor) resulting in elevated secretion of hexosaminidase. In agreement with this, our studies identified novel host GCC185 protein that interacts with Nef EEEE65 acidic domain interfering GCC185-Rab9 vesicle membrane fusion responsible for retrograde vesicular transport of CI-MPR from late endosomes to TGN. In light of existing report suggesting critical role of Nef-GCC185 interaction reveals valuable mechanistic insights affecting specific protein transport pathway in docking of late endosome derived Rab9 bearing transport vesicle at TGN elucidating role of Nef during viral pathogenesis. -- Highlights: •Nef, an accessory protein of HIV-1 interacts with host factor and culminates into AIDS pathogenesis. •Using Caenorhabditis elegans based screen system, novel Nef interacting cellular protein GCC185 was identified. •Molecular characterization of Nef and human protein GCC185 revealed Nef EEEE{sub 65} key region interacted with full length GCC185. •Nef impeded the GCC185-Rab 9 interaction and

Solo/Trio8, a membrane-associated short isoform of Trio, modulates endosome dynamics and neurite elongation.

Science.gov (United States)

Sun, Ying-Jie; Nishikawa, Kaori; Yuda, Hideki; Wang, Yu-Lai; Osaka, Hitoshi; Fukazawa, Nobuna; Naito, Akira; Kudo, Yoshihisa; Wada, Keiji; Aoki, Shunsuke

2006-09-01

With DNA microarrays, we identified a gene, termed Solo, that is downregulated in the cerebellum of Purkinje cell degeneration mutant mice. Solo is a mouse homologue of rat Trio8-one of multiple Trio isoforms recently identified in rat brain. Solo/Trio8 contains N-terminal sec14-like and spectrin-like repeat domains followed by a single guanine nucleotide exchange factor 1 (GEF1) domain, but it lacks the C-terminal GEF2, immunoglobulin-like, and kinase domains that are typical of Trio. Solo/Trio8 is predominantly expressed in Purkinje neurons of the mouse brain, and expression begins following birth and increases during Purkinje neuron maturation. We identified a novel C-terminal membrane-anchoring domain in Solo/Trio8 that is required for enhanced green fluorescent protein-Solo/Trio8 localization to early endosomes (positive for both early-endosome antigen 1 [EEA1] and Rab5) in COS-7 cells and primary cultured neurons. Solo/Trio8 overexpression in COS-7 cells augmented the EEA1-positive early-endosome pool, and this effect was abolished via mutation and inactivation of the GEF domain or deletion of the C-terminal membrane-anchoring domain. Moreover, primary cultured neurons transfected with Solo/Trio8 showed increased neurite elongation that was dependent on these domains. These results suggest that Solo/Trio8 acts as an early-endosome-specific upstream activator of Rho family GTPases for neurite elongation of developing Purkinje neurons.

Impaired degradation followed by enhanced recycling of epidermal growth factor receptor caused by hypo-phosphorylation of tyrosine 1045 in RBE cells

International Nuclear Information System (INIS)

Gui, Anping; Kobayashi, Akira; Motoyama, Hiroaki; Kitazawa, Masato; Takeoka, Michiko; Miyagawa, Shinichi

2012-01-01

Since cholangiocarcinoma has a poor prognosis, several epidermal growth factor receptor (EGFR)-targeted therapies with antibody or small molecule inhibitor treatment have been proposed. However, their effect remains limited. The present study sought to understand the molecular genetic characteristics of cholangiocarcinoma related to EGFR, with emphasis on its degradation and recycling. We evaluated EGFR expression and colocalization by immunoblotting and immunofluorescence, cell surface EGFR expression by fluorescence-activated cell sorting (FACS), and EGFR ubiquitination and protein binding by immunoprecipitation in the human cholangiocarcinoma RBE and immortalized cholangiocyte MMNK-1 cell lines. Monensin treatment and Rab11a depletion by siRNA were adopted for inhibition of EGFR recycling. Upon stimulation with EGF, ligand-induced EGFR degradation was impaired and the expression of phospho-tyrosine 1068 and phospho-p44/42 MAPK was sustained in RBE cells as compared with MMNK-1 cells. In RBE cells, the process of EGFR sorting for lysosomal degradation was blocked at the early endosome stage, and non-degradated EGFR was recycled to the cell surface. A disrupted association between EGFR and the E3 ubiquitin ligase c-Cbl, as well as hypo-phosphorylation of EGFR at tyrosine 1045 (Tyr1045), were also observed in RBE cells. In RBE cells, up-regulation of EGFR Tyr1045 phosphorylation is a potentially useful molecular alteration in EGFR-targeted therapy. The combination of molecular-targeted therapy determined by the characteristics of individual EGFR phosphorylation events and EGFR recycling inhibition show promise in future treatments of cholangiocarcinoma

FY1998 research report on the R and D on recycling technology. Part 1; 1998 nendo recycle nado kankyo gijutsu kekyu kaihatsu seika hokokusho. 1

Energy Technology Data Exchange (ETDEWEB)

NONE

1999-03-01

This project aims to develop recycling technology for reduction of environmental burden caused by waste, and promotion of recycling of wastes. As for advanced recycling technology for PET bottles, the facility was improved for improvement of a facility operability and product quality. Study was made on the effect of a raw bale quality and recycled flake colors on a product quality, the forming test with PS or PET labels, and the concentration and effect of washing liquid circulated in flake washing process. As for recycling technology of hard-to-dispose waste plastics, facility improvement and demonstration test were made for continuous operation of dry-distillation/gasification of shredder dusts and separation of nonferrous metals and glass. Study was also made on pulverizing and recycling technologies of FRP bath tubs, and such pulverization costs were estimated. As for production technology of chemical feed and fuel from wastes, study was made on removal technology of non-flammable substances, development of alkaline additives, reacting condition, development of reactors, and use technology as chemical feed. (NEDO)

Households’ Perception of Financial Incentives in Endorsing Sustainable Waste Recycling in Nigeria

Directory of Open Access Journals (Sweden)

Beatrice Abila

2018-06-01

Full Text Available Recycling is viewed as a central aspect in sustainability and mainly as pro-environmental consumer behavior. The purpose of this study is to examine the perception of households on financial incentives in endorsing sustainable recycling for municipal solid waste in Nigeria. The study was conducted in the Shomolu Local Government Area, Lagos State, Nigeria. The study also covers drivers for household willingness to recycle municipal solid waste on environmental risk, behavioral economics, resource value, economic benefit, convenience, knowledge, legislation, and belief. The result from the study asserts the hypothesis that financial incentives for recycling are vital for reducing and managing municipal solid waste sustainably. The most important driver for household willingness to recycle municipal solid waste is the detrimental environmental impacts. A moderate to positive relationship exists between households’ perception of financial incentives for recycling and drivers for household willingness to recycle municipal solid waste. The study recommends adopting the extended producer responsibility (EPR model, reverse vending options, amongst other approaches, in an effort to promote recycling culture among citizens and residents in Nigeria.

Recycling as a strategy against rare earth element criticality: a systemic evaluation of the potential yield of NdFeB magnet recycling.

Science.gov (United States)

Rademaker, Jelle H; Kleijn, René; Yang, Yongxiang

2013-09-17

End-of-life recycling is promoted by OECD countries as a promising strategy in the current global supply crisis surrounding rare earth elements (REEs) so that dependence on China, the dominant supplier, can be decreased. So far the feasibility and potential yield of REE recycling has not been systematically evaluated. This paper estimates the annual waste flows of neodymium and dysprosium from permanent magnets, the main deployment of these critical REEs, during the 2011-2030 period. The estimates focus on three key permanent magnet waste flows: wind turbines, hybrid and electric vehicles, and hard disk drives (HDDs) in personal computers (PCs). This is a good indication of the end-of-life recycling of neodymium and dysprosium maximum potential yield. Results show that for some time to come, waste flows from permanent magnets will remain small relative to the rapidly growing global REE demand. Policymakers therefore need to be aware that during the next decade recycling is unlikely to substantially contribute to global REE supply security. In the long term, waste flows will increase sharply and will meet a substantial part of the total demand for these metals. Future REE recycling efforts should, therefore, focus on the development of recycling technology and infrastructure.

RCP-driven α5β1 recycling suppresses Rac and promotes RhoA activity via the RacGAP1–IQGAP1 complex

Science.gov (United States)

Jacquemet, Guillaume; Green, David M.; Bridgewater, Rebecca E.; von Kriegsheim, Alexander; Humphries, Martin J.; Norman, Jim C.

2013-01-01

Inhibition of αvβ3 or expression of mutant p53 promotes invasion into fibronectin (FN)-containing extracellular matrix (ECM) by enhancing Rab-coupling protein (RCP)–dependent recycling of α5β1 integrin. RCP and α5β1 cooperatively recruit receptor tyrosine kinases, including EGFR1, to regulate their trafficking and downstream signaling via protein kinase B (PKB)/Akt, which, in turn, promotes invasive migration. In this paper, we identify a novel PKB/Akt substrate, RacGAP1, which is phosphorylated as a consequence of RCP-dependent α5β1 trafficking. Phosphorylation of RacGAP1 promotes its recruitment to IQGAP1 at the tips of invasive pseudopods, and RacGAP1 then locally suppresses the activity of the cytoskeletal regulator Rac and promotes the activity of RhoA in this subcellular region. This Rac to RhoA switch promotes the extension of pseudopodial processes and invasive migration into FN-containing matrices, in a RhoA-dependent manner. Thus, the localized endocytic trafficking of α5β1 within the tips of invasive pseudopods elicits signals that promote the reorganization of the actin cytoskeleton, protrusion, and invasion into FN-rich ECM. PMID:24019536

ABMA, a small molecule that inhibits intracellular toxins and pathogens by interfering with late endosomal compartments.

Science.gov (United States)

Wu, Yu; Pons, Valérie; Goudet, Amélie; Panigai, Laetitia; Fischer, Annette; Herweg, Jo-Ana; Kali, Sabrina; Davey, Robert A; Laporte, Jérôme; Bouclier, Céline; Yousfi, Rahima; Aubenque, Céline; Merer, Goulven; Gobbo, Emilie; Lopez, Roman; Gillet, Cynthia; Cojean, Sandrine; Popoff, Michel R; Clayette, Pascal; Le Grand, Roger; Boulogne, Claire; Tordo, Noël; Lemichez, Emmanuel; Loiseau, Philippe M; Rudel, Thomas; Sauvaire, Didier; Cintrat, Jean-Christophe; Gillet, Daniel; Barbier, Julien

2017-11-14

Intracellular pathogenic microorganisms and toxins exploit host cell mechanisms to enter, exert their deleterious effects as well as hijack host nutrition for their development. A potential approach to treat multiple pathogen infections and that should not induce drug resistance is the use of small molecules that target host components. We identified the compound 1-adamantyl (5-bromo-2-methoxybenzyl) amine (ABMA) from a cell-based high throughput screening for its capacity to protect human cells and mice against ricin toxin without toxicity. This compound efficiently protects cells against various toxins and pathogens including viruses, intracellular bacteria and parasite. ABMA provokes Rab7-positive late endosomal compartment accumulation in mammalian cells without affecting other organelles (early endosomes, lysosomes, the Golgi apparatus, the endoplasmic reticulum or the nucleus). As the mechanism of action of ABMA is restricted to host-endosomal compartments, it reduces cell infection by pathogens that depend on this pathway to invade cells. ABMA may represent a novel class of broad-spectrum compounds with therapeutic potential against diverse severe infectious diseases.

On recycling of nuclear fuel in Japan

International Nuclear Information System (INIS)

1992-01-01

In Japan, atomic energy has become to accomplish the important role in energy supply. Recently the interest in the protection of global environment heightened, and the anxiety on oil supply has been felt due to the circumstances in Mideast. Therefore, the importance of atomic energy as an energy source for hereafter increased, and the future plan of nuclear fuel recycling in Japan must be promoted on such viewpoint. At present in Japan, the construction of nuclear fuel cycle facilities is in progress in Rokkasho, Aomori Prefecture. The prototype FBR 'Monju' started the general functional test in May, this year. The transport of the plutonium reprocessed in U.K. and France to Japan will be carried out in near future. This report presents the concrete measures of nuclear fuel recycling in Japan from the long term viewpoint up to 2010. The necessity and meaning of nuclear fuel recycling in Japan, the effort related to nuclear nonproliferation, the plan of nuclear fuel recycling for hereafter in Japan, the organization of MOX fuel fabrication in Japan and abroad, the method of utilizing recovered uranium and the reprocessing of spent MOX fuel are described. (K.I.)

Altered neurological function in mice immunized with early endosome antigen 1

Directory of Open Access Journals (Sweden)

Fritzler Marvin J

2004-01-01

Full Text Available Abstract Background Autoantibodies directed against the 160 kDa endosome protein early endosome antigen 1 (EEA1 are seen in patients with neurological diseases. To determine if antibodies to EEA1 have a neuropathological effect, mice from three major histocompatability haplotype backgrounds (H2q, H2b and H2d were immunized with EEA1 (amino acids 82–1411 that was previously shown to contain the target EEA1 epitopes. The mice were then subjected to five neuro-behavioural tests: grid walking, forelimb strength, open field, reaching and rotarod. Results The immunized SWR/J mice with sustained anti-EEA1 antibodies had significantly reduced forelimb strength than the control non-immune mice of the same strain, and BALB/CJ immune mice demonstrated significantly more forelimb errors on the grid walk test than the control group. Conclusions Antibodies to recombinant EEA1 in mice may mediate neurological deficits that are consistent with clinical features of some humans that spontaneously develop anti-EEA1 autoantibodies.

The importance of recycling - Responsible recycling

International Nuclear Information System (INIS)

Svensson, Joens Petter

2014-01-01

7 times the total emissions from Sweden are saved each year by the recycling industry. It reduces CO 2 emissions and saves the environment. In fact it annually reduces global CO 2 emissions by 500 million tons, which is more than what is being emitted by the world wide aviation industry. Recycling of iron and steel saves 74% energy and reduces water and air pollution by respectively 76% and 86%, compared to primary production. It provides new raw materials and contributes to save energy. There's no sense in producing goods in a permanent material like plastics, that's supposed to be used only once. It's a huge waste of resources. Today the recycling industry provides half of the world's raw materials and this figure is set to increase. It's about environmentally sound management of resources. It's about plain common sense. There has to be a political willingness to facilitate recycling in every way. And from a corporate perspective social responsibility is becoming an increasingly important competitive edge. This is also a communication issue, it has to be a fact that is well known to the market when a company is doing valuable environmental work. We also need a well functioning global market with easy to understand regulations to facilitate global trade. The global demand for recycled materials should influence their collection and use. Fraud and theft has also to be kept at bay which calls for a close collaboration between organizations such as The International Chamber of Commerce, The International Trade Council and the International Maritime Bureau of the commercial crime services. Increasing recycling is the only way to go if we want to minimize our effect on the environment. We have to remember that recycling is essential for the environment. An increase would be a tremendous help to reduce the green house effect. Increasing recycling is not rocket science. We know how to do it, we just have to decide to go through with it

Recycling Facilities - Land Recycling Cleanup Locations

Data.gov (United States)

NSGIC Education | GIS Inventory — Land Recycling Cleanup Location Land Recycling Cleanup Locations (LRCL) are divided into one or more sub-facilities categorized as media: Air, Contained Release or...

The NKG2D ligand ULBP2 is specifically regulated through an invariant chain-dependent endosomal pathway

DEFF Research Database (Denmark)

Uhlenbrock, Franziska Katharina; Hagemann-Jensen, Michael Henrik; Kehlet, Stephanie

2014-01-01

by affecting endosomal/lysosomal integrity and protein kinase C activity. The invariant chain was further essential for endosomal transport of ULBP2. This novel pathway was identified through screening experiments by which methylselenic acid was found to possess notable NKG2D ligand regulatory properties....... The protein kinase C inhibitor methylselenic acid induced MICA/B surface expression but dominantly blocked ULBP2 surface transport. Remarkably, by targeting this novel pathway we could specifically block the production of soluble ULBP2 from different, primary melanomas. Our findings strongly suggest...

Requirement analysis to promote small-sized E-waste collection from consumers.

Science.gov (United States)

Mishima, Kuniko; Nishimura, Hidekazu

2016-02-01

The collection and recycling of small-sized waste electrical and electronic equipment is an emerging problem, since these products contain certain amounts of critical metals and rare earths. Even if the amount is not large, having a few supply routes for such recycled resources could be a good strategy to be competitive in a world of finite resources. The small-sized e-waste sometimes contains personal information, therefore, consumers are often reluctant to put them into recycling bins. In order to promote the recycling of E-waste, collection of used products from the consumer becomes important. Effective methods involving incentives for consumers might be necessary. Without such methods, it will be difficult to achieve the critical amounts necessary for an efficient recycling system. This article focused on used mobile phones among information appliances as the first case study, since it contains relatively large amounts of valuable metals compared with other small-sized waste electrical and electronic equipment and there are a large number of products existing in the market. The article carried out surveys to determine what kind of recycled material collection services are preferred by consumers. The results clarify that incentive or reward money alone is not a driving force for recycling behaviour. The article discusses the types of effective services required to promote recycling behaviour. The article concludes that securing information, transferring data and providing proper information about resources and environment can be an effective tool to encourage a recycling behaviour strategy to promote recycling, plus the potential discount service on purchasing new products associated with the return of recycled mobile phones. © The Author(s) 2015.

Nuclear recycling

International Nuclear Information System (INIS)

Spinrad, B.I.

1985-01-01

This paper discusses two aspects of the economics of recycling nuclear fuel: the actual costs and savings of the recycling operation in terms of money spent, made, and saved; and the impact of the recycling on the future cost of uranium. The authors review the relevant physical and chemical processes involved in the recycling process. Recovery of uranium and plutonium is discussed. Fuel recycling in LWRs is examined and a table presents the costs of reprocessing and not reprocessing. The subject of plutonium in fast reactors is addressed. Safeguards and weapons proliferation are discussed

Kinesin Khc-73/KIF13B modulates retrograde BMP signaling by influencing endosomal dynamics at the Drosophila neuromuscular junction.

Science.gov (United States)

Liao, Edward H; Gray, Lindsay; Tsurudome, Kazuya; El-Mounzer, Wassim; Elazzouzi, Fatima; Baim, Christopher; Farzin, Sarah; Calderon, Mario R; Kauwe, Grant; Haghighi, A Pejmun

2018-01-01

Retrograde signaling is essential for neuronal growth, function and survival; however, we know little about how signaling endosomes might be directed from synaptic terminals onto retrograde axonal pathways. We have identified Khc-73, a plus-end directed microtubule motor protein, as a regulator of sorting of endosomes in Drosophila larval motor neurons. The number of synaptic boutons and the amount of neurotransmitter release at the Khc-73 mutant larval neuromuscular junction (NMJ) are normal, but we find a significant decrease in the number of presynaptic release sites. This defect in Khc-73 mutant larvae can be genetically enhanced by a partial genetic loss of Bone Morphogenic Protein (BMP) signaling or suppressed by activation of BMP signaling in motoneurons. Consistently, activation of BMP signaling that normally enhances the accumulation of phosphorylated form of BMP transcription factor Mad in the nuclei, can be suppressed by genetic removal of Khc-73. Using a number of assays including live imaging in larval motor neurons, we show that loss of Khc-73 curbs the ability of retrograde-bound endosomes to leave the synaptic area and join the retrograde axonal pathway. Our findings identify Khc-73 as a regulator of endosomal traffic at the synapse and modulator of retrograde BMP signaling in motoneurons.

Disposing and recycling waste printed circuit boards: disconnecting, resource recovery, and pollution control.

Science.gov (United States)

Wang, Jianbo; Xu, Zhenming

2015-01-20

Over the past decades, China has been suffering from negative environmental impacts from distempered e-waste recycling activities. After a decade of effort, disassembly and raw materials recycling of environmentally friendly e-waste have been realized in specialized companies, in China, and law enforcement for illegal activities of e-waste recycling has also been made more and more strict. So up to now, the e-waste recycling in China should be developed toward more depth and refinement to promote industrial production of e-waste resource recovery. Waste printed circuit boards (WPCBs), which are the most complex, hazardous, and valuable components of e-waste, are selected as one typical example in this article that reviews the status of related regulations and technologies of WPCBs recycling, then optimizes, and integrates the proper approaches in existence, while the bottlenecks in the WPCBs recycling system are analyzed, and some preliminary experiments of pinch technologies are also conducted. Finally, in order to provide directional guidance for future development of WPCBs recycling, some key points in the WPCBs recycling system are proposed to point towards a future trend in the e-waste recycling industry.

Molecular determinants of the interaction between Doa1 and Hse1 involved in endosomal sorting.

Science.gov (United States)

Han, Seungsu; Shin, Donghyuk; Choi, Hoon; Lee, Sangho

2014-03-28

Yeast Doa1/Ufd3 is an adaptor protein for Cdc48 (p97 in mammal), an AAA type ATPase associated with endoplasmic reticulum-associated protein degradation pathway and endosomal sorting into multivesicular bodies. Doa1 functions in the endosomal sorting by its association with Hse1, a component of endosomal sorting complex required for transport (ESCRT) system. The association of Doa1 with Hse1 was previously reported to be mediated between PFU domain of Doa1 and SH3 of Hse1. However, it remains unclear which residues are specifically involved in the interaction. Here we report that Doa1/PFU interacts with Hse1/SH3 with a moderate affinity of 5 μM. Asn-438 of Doa1/PFU and Trp-254 of Hse1/SH3 are found to be critical in the interaction while Phe-434, implicated in ubiquitin binding via a hydrophobic interaction, is not. Small-angle X-ray scattering measurements combined with molecular docking and biochemical analysis yield the solution structure of the Doa1/PFU:Hse1/SH3 complex. Taken together, our results suggest that hydrogen bonding is a major determinant in the interaction of Doa1/PFU with Hse1/SH3. Copyright © 2014 Elsevier Inc. All rights reserved.

Molecular basis for SNX-BAR-mediated assembly of distinct endosomal sorting tubules

DEFF Research Database (Denmark)

van Weering, Jan R.T.; Sessions, Richard B.; Traer, Colin J.

2012-01-01

that dimerization is achieved in part through neutralization of charged residues in the hydrophobic BAR-dimerization interface. Membrane remodelling also requires functional amphipathic helices, predicted to be present in all SNX-BARs, and the formation of high order SNX-BAR oligomers through selective 'tip...... and organizes the tubular endosomal network....

Augmented internalisation of ferroportin to late endosomes impairs iron uptake by enterocyte-like IEC-6 cells.

Science.gov (United States)

Oates, Phillip S; Thomas, Carla

2005-08-01

Absorption of iron occurs by duodenal enterocytes, involving uptake by the divalent metal transporter-1 (DMT1) and release by ferroportin. Ferroportin responds to the hepatocyte-produced 25-amino-acid-peptide hepcidin-25 by undergoing internalisation to late endosomes that impair iron release. Ferroportin is also expressed on the apical membrane of polarised Caco-2 cells, rat intestinal cells and in IEC-6 cells (an intestinal epithelial cell line). A blocking antibody to ferroportin also impairs the uptake, but not the release, of iron. In this study IEC-6 cells were used to study the mechanism of impairment or recovery from impairment produced by the blocking antibody and the fate of DMT1 and ferroportin. Uptake of 1 muM Fe(II) was studied by adding the antibody from time 0 and after adding or removing the antibody once a steady state had been reached. Surface binding, maximum iron transport rate V(max) and transporter affinity (K(m)) were measured after impairment of iron uptake. Ferroportin and DMT1 distribution were assessed by immunofluorescence microscopy. Antibody-mediated impairment, or recovery from impairment, of Fe(II) uptake occurred within minutes. Impairment was lost when the antibody was combined with the immunizing peptide. DMT1 and ferroportin undergo internalisation to late endosomes and, in the presence of the antibody, augmented internalisation of DMT1 and ferroportin caused swelling of late endosomes. Surface binding of Fe(II) and iron transport V(max) were reduced by 50%, indicating that the antibody removed membrane-bound DMT1. The ferroportin antibody induced rapid turnover of membrane ferroportin and DMT1 and its internalisation to late endosomes, resulting in impaired Fe(II) uptake.

Comparisons of four categories of waste recycling in China’s paper industry based on physical input–output life-cycle assessment model

International Nuclear Information System (INIS)

Liang Sai; Zhang, Tianzhu; Xu Yijian

2012-01-01

Highlights: ► Using crop straws and wood wastes for paper production should be promoted. ► Bagasse and textile waste recycling should be properly limited. ► Imports of scrap paper should be encouraged. ► Sensitivity analysis, uncertainties and policy implications are discussed. - Abstract: Waste recycling for paper production is an important component of waste management. This study constructs a physical input–output life-cycle assessment (PIO-LCA) model. The PIO-LCA model is used to investigate environmental impacts of four categories of waste recycling in China’s paper industry: crop straws, bagasse, textile wastes and scrap paper. Crop straw recycling and wood utilization for paper production have small total intensity of environmental impacts. Moreover, environmental impacts reduction of crop straw recycling and wood utilization benefits the most from technology development. Thus, using crop straws and wood (including wood wastes) for paper production should be promoted. Technology development has small effects on environmental impacts reduction of bagasse recycling, textile waste recycling and scrap paper recycling. In addition, bagasse recycling and textile waste recycling have big total intensity of environmental impacts. Thus, the development of bagasse recycling and textile waste recycling should be properly limited. Other pathways for reusing bagasse and textile wastes should be explored and evaluated. Moreover, imports of scrap paper should be encouraged to reduce large indirect impacts of scrap paper recycling on domestic environment.

Acid-sensing ion channel (ASIC) 4 predominantly localizes to an early endosome-related organelle upon heterologous expression.

Science.gov (United States)

Schwartz, Verena; Friedrich, Katharina; Polleichtner, Georg; Gründer, Stefan

2015-12-15

Acid-sensing ion channels (ASICs) are voltage-independent proton-gated amiloride sensitive sodium channels, belonging to the DEG/ENaC gene family. Six different ASICs have been identified (ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3, ASIC4) that are activated by a drop in extracellular pH, either as homo- or heteromers. An exception is ASIC4, which is not activated by protons as a homomer and which does not contribute to functional heteromeric ASICs. Insensitivity of ASIC4 to protons and its comparatively low sequence identity to other ASICs (45%) raises the question whether ASIC4 may have different functions than other ASICs. In this study, we therefore investigated the subcellular localization of ASIC4 in heterologous cell lines, which revealed a surprising accumulation of the channel in early endosome-related vacuoles. Moreover, we identified an unique amino-terminal motif as important for forward-trafficking from the ER/Golgi to the early endosome-related compartment. Collectively, our results show that heterologously expressed ASIC4 predominantly resides in an intracellular endosomal compartment.

Investigations on Fresh and Hardened Properties of Recycled Aggregate Self Compacting Concrete

Science.gov (United States)

Revathi, P.; Selvi, R. S.; Velin, S. S.

2013-09-01

In the recent years, construction and demolition waste management issues have attracted the attention from researchers around the world. In the present study, the potential usage of recycled aggregate obtained from crushed demolition waste for making self compacting concrete (SCC) was researched. The barriers in promoting the use of recycled material in new construction are also discussed. In addition, the results of an experimental study involving the use of recycled concrete aggregate as coarse aggregates for producing self-compacting concrete to study their flow and strength characteristics are also presented. Five series of mixture were prepared with 0, 25, 50, 75, and 100 % coarse recycled aggregate adopting Nan Su's mix proportioning method. The fresh concrete properties were evaluated through the slump flow, J-ring and V-funnel tests. Compressive and tensile strengths were also determined. The results obtained showed that SCC could be successfully developed by incorporating recycled aggregates.

Modelling Recycling Targets

DEFF Research Database (Denmark)

Hill, Amanda Louise; Leinikka Dall, Ole; Andersen, Frits M.

2014-01-01

Within the European Union (EU) a paradigm shift is currently occurring in the waste sector, where EU waste directives and national waste strategies are placing emphasis on resource efficiency and recycling targets. The most recent Danish resource strategy calculates a national recycling rate of 22......% for household waste, and sets an ambitious goal of a 50% recycling rate by 2020. This study integrates the recycling target into the FRIDA model to project how much waste and from which streams should be diverted from incineration to recycling in order to achieve the target. Furthermore, it discusses how...... the existing technological, organizational and legislative frameworks may affect recycling activities. The results of the analysis show that with current best practice recycling rates, the 50% recycling rate cannot be reached without recycling of household biowaste. It also shows that all Danish municipalities...

Unconventional recycling

Energy Technology Data Exchange (ETDEWEB)

White, K.M.

1996-05-01

Despite advances made in recycling technology and markets for materials over the past few years, recycling at convention centers, particularly on the show floor itself, can be a vexing problem. Part of the problem lies in the fact that recycling at convention centers has more to do with logistics than it does with these industry trends. However, given the varied nature of convention centers, and the shows they book, a rigid approach to recycling at convention centers is not always feasible. Like the numerous different curbside programs serving communities across the country, what works for one convention center--and one show--many not work for another. These difficulties notwithstanding, more convention centers are offering recycling programs today, and more groups booking conventions these days have begun requesting recycling services.

Recycling Lesson Plan

Science.gov (United States)

Okaz, Abeer Ali

2013-01-01

This lesson plan designed for grade 2 students has the goal of teaching students about the environmental practice of recycling. Children will learn language words related to recycling such as: "we can recycle"/"we can't recycle" and how to avoid littering with such words as: "recycle paper" and/or "don't throw…

Research on the Phenomenon of Chinese Residents’ Spiritual Contagion for the Reuse of Recycled Water Based on SC-IAT

Directory of Open Access Journals (Sweden)

Hanliang Fu

2017-11-01

Full Text Available Recycled water has been widely recognized in the world as an effective approach to relieve the issue of water shortage. Meanwhile, with several decades of development, the insufficiency of technology is no longer the primary factor that restricts the popularization of recycled water. What makes it difficult to promote the concept of reusing recycled water in China? To solve this issue, a special experiment on the public’s attitude towards the reuse of recycled water was designed based on a Single Category Implicit Association Test (SC-IAT, so as to avoid factors like social preference that can influence the survey results, and to gain the public’s negative implicit attitude towards reusing recycled water reuse, which is close to the public’s real attitude to it. From the perspective of implicit attitude, this research testifies the “spiritual contagion” phenomenon of the public, which refers to refusing recycled water reuse because recycled water is made from sewage treatment. By comparing the implicit attitude to recycled water reuse with the explicit attitude that is acquired from self-reporting questionnaires about reusing recycled water, this research finds that the implicit attitude is more positive than the explicit attitude, which accounts for the phenomenon of “best game no one played” in the promotion of the recycled water reuse, that is, the public though applauding the environment-friendly policy, will not actually use the recycled water.

Waste reduction and recycling initiatives in Japanese cities: lessons from Yokohama and Kamakura.

Science.gov (United States)

Hotta, Yasuhiko; Aoki-Suzuki, Chika

2014-09-01

Waste reduction and recycling at the city level will acquire greater significance in the near future due to rising global volumes of waste. This paper seeks to identify policy-relevant drivers for successful promotion of waste reduction and recycling. Factors influencing the success of waste reduction and recycling campaigns are identified. Two case study cities in Japan which depict the successful use of the 3Rs (reduce, reuse and recycle) at the municipal level are presented. In these cases, the existence of incinerators, which are generally considered as disincentives for recycling, was not functioning as a disincentive but rather as an incentive for waste reduction. Owing to the high cost of incineration facilities, the movement to close incinerators has become a strong incentive for waste reduction and recycling in these two cities. The study suggests that careful consideration is necessary when making decisions concerning high-cost waste treatment facilities with high installation, maintenance and renewal outlays. In addition, intensive source separation and other municipal recycling initiatives have a high potential for producing positive results. © The Author(s) 2014.

Waste material recycling: Assessment of contaminants limiting recycling

DEFF Research Database (Denmark)

Pivnenko, Kostyantyn

systematically investigated. This PhD project provided detailed quantitative data following a consistent approach to assess potential limitations for the presence of chemicals in relation to material recycling. Paper and plastics were used as illustrative examples of materials with well-established recycling...... schemes and great potential for increase in recycling, respectively. The approach followed in the present work was developed and performed in four distinct steps. As step one, fractional composition of waste paper (30 fractions) and plastics (9 fractions) from households in Åbenrå municipality (Southern...... detrimental to their recycling. Finally, a material flow analysis (MFA) approach revealed the potential for accumulation and spreading of contaminants in material recycling, on the example of the European paper cycle. Assessment of potential mitigation measures indicated that prevention of chemical use...

The potential environmental gains from recycling waste plastics: Simulation of transferring recycling and recovery technologies to Shenyang, China

International Nuclear Information System (INIS)

Chen Xudong; Xi Fengming; Geng Yong; Fujita, Tsuyoshi

2011-01-01

Research highlights: → Urban symbiosis creates compatibility of industrial development and waste management. → Mechanical technology leads to more CO 2 emission reduction. → Energy recovery technology leads to more fossil fuel saving. → Clean energy makes recycling technologies cleaner. → Demand management is crucial for realizing potential environmental gains of recycling. - Abstract: With the increasing attention on developing a low-carbon economy, it is necessary to seek appropriate ways on reducing greenhouse gas (GHG) emissions through innovative municipal solid waste management (MSWM), such as urban symbiosis. However, quantitative assessments on the environmental benefits of urban symbiosis, especially in developing countries, are limited because only a limited number of planned synergistic activities have been successful and it is difficult to acquire detailed inventory data from private companies. This paper modifies and applies a two-step simulation system and used it to assess the potential environmental benefits, including the reduction of GHG emissions and saving of fossil fuels, by employing various Japanese plastics recycling/energy-recovery technologies in Shenyang, China. The results showed that among various recycling/energy-recovery technologies, the mechanical waste plastics recycling technology, which produces concrete formwork boards (NF boards), has the greatest potential in terms of reducing GHG emissions (1.66 kg CO 2 e/kg plastics), whereas the technology for the production of refuse plastic fuel (RPF) has the greatest potential on saving fossil fuel consumption (0.77 kgce/kg-plastics). Additional benefits can be gained by applying combined technologies that cascade the utilization of waste plastics. Moreover, the development of clean energy in conjunction with the promotion of new waste plastics recycling programs could contribute to additional reductions in GHG emissions and fossil fuel consumption.

Role of the AP-5 adaptor protein complex in late endosome-to-Golgi retrieval.

Directory of Open Access Journals (Sweden)

Jennifer Hirst

2018-01-01

Full Text Available The AP-5 adaptor protein complex is presumed to function in membrane traffic, but so far nothing is known about its pathway or its cargo. We have used CRISPR-Cas9 to knock out the AP-5 ζ subunit gene, AP5Z1, in HeLa cells, and then analysed the phenotype by subcellular fractionation profiling and quantitative mass spectrometry. The retromer complex had an altered steady-state distribution in the knockout cells, and several Golgi proteins, including GOLIM4 and GOLM1, were depleted from vesicle-enriched fractions. Immunolocalisation showed that loss of AP-5 led to impaired retrieval of the cation-independent mannose 6-phosphate receptor (CIMPR, GOLIM4, and GOLM1 from endosomes back to the Golgi region. Knocking down the retromer complex exacerbated this phenotype. Both the CIMPR and sortilin interacted with the AP-5-associated protein SPG15 in pull-down assays, and we propose that sortilin may act as a link between Golgi proteins and the AP-5/SPG11/SPG15 complex. Together, our findings suggest that AP-5 functions in a novel sorting step out of late endosomes, acting as a backup pathway for retromer. This provides a mechanistic explanation for why mutations in AP-5/SPG11/SPG15 cause cells to accumulate aberrant endolysosomes, and highlights the role of endosome/lysosome dysfunction in the pathology of hereditary spastic paraplegia and other neurodegenerative disorders.

Ebola Viral Glycoprotein Bound to Its Endosomal Receptor Niemann-Pick C1.

Science.gov (United States)

Wang, Han; Shi, Yi; Song, Jian; Qi, Jianxun; Lu, Guangwen; Yan, Jinghua; Gao, George F

2016-01-14

Filoviruses, including Ebola and Marburg, cause fatal hemorrhagic fever in humans and primates. Understanding how these viruses enter host cells could help to develop effective therapeutics. An endosomal protein, Niemann-Pick C1 (NPC1), has been identified as a necessary entry receptor for this process, and priming of the viral glycoprotein (GP) to a fusion-competent state is a prerequisite for NPC1 binding. Here, we have determined the crystal structure of the primed GP (GPcl) of Ebola virus bound to domain C of NPC1 (NPC1-C) at a resolution of 2.3 Å. NPC1-C utilizes two protruding loops to engage a hydrophobic cavity on head of GPcl. Upon enzymatic cleavage and NPC1-C binding, conformational change in the GPcl further affects the state of the internal fusion loop, triggering membrane fusion. Our data therefore provide structural insights into filovirus entry in the late endosome and the molecular basis for design of therapeutic inhibitors of viral entry. Copyright © 2016 Elsevier Inc. All rights reserved.

Vps33B is required for delivery of endocytosed cargo to lysosomes

NARCIS (Netherlands)

Galmes, Romain; ten Brink, Corlinda; Oorschot, Viola; Veenendaal, Tineke; Jonker, Caspar; van der Sluijs, Peter; Klumperman, Judith

2015-01-01

In mammalian cells Vps33B forms a complex with VIPAS-39 that is recruited to recycling endosomes. Here we show that when Vps33B is expressed together with Rab7-interacting lysosomal protein (RILP) it is recruited to late endosomes-lysosomes and that depletion of Vps33B impairs late

On achieving the state's household recycling target: A case study of Northern New Jersey, USA

International Nuclear Information System (INIS)

Otegbeye, M.; Abdel-Malek, L.; Hsieh, H.N.; Meegoda, J.N.

2009-01-01

In recent times, the State of New Jersey (USA) has been making attempts at promoting recycling as an environmentally friendly means of attaining self-sufficiency at waste disposal, and the state has put in place a 50% recycling target for its municipal solid waste stream. While the environmental benefits of recycling are obvious, a recycling program must be cost effective to ensure its long-term sustainability. In this paper, a linear programming model is developed to examine the current state of recycling in selected counties in Northern New Jersey and assess the needs to achieve the state's recycling goal in these areas. The optimum quantities of waste to be sent to the different waste facilities, which include landfills, incinerators, transfer stations, recycling and composting plants, are determined by the model. The study shows that for these counties, the gap between the current waste practices where the recycling rate stands at 32% and the state's goal can be bridged by more efficient utilization of existing facilities and reasonable investment in expanding those for recycling activities

Decoupling internalization, acidification and phagosomal-endosomal/lysosomal fusion during phagocytosis of InlA coated beads in epithelial cells.

Directory of Open Access Journals (Sweden)

Craig D Blanchette

Full Text Available BACKGROUND: Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells. Therefore, in this study, we developed a simple method to measure and decouple particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK and Caco-2 epithelial cells. METHODOLOGY/PRINCIPAL FINDINGS: Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA, a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated phagocytosis. We were able to independently measure the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads with antibody quenching, a pH sensitive dye and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we exploited the phagosomal acidification process to demonstrate an additional, real-time method for tracking bead binding, internalization and phagosomal acidification. CONCLUSIONS/SIGNIFICANCE: Using this method, we found that the time scales for internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion ranged from 23-32 min, 3-4 min and 74-120 min, respectively, for MDCK and Caco-2 epithelial cells. Both the static and real-time methods developed here are expected to be readily and broadly applicable, as they simply

Municipal recycling support program. Guide to applicants

Energy Technology Data Exchange (ETDEWEB)

1989-01-01

The Municipal Recycling Support Program stems from the Ontario Ministry of the Environment's policies and programs begun in 1980 aimed at encouraging the development of source separation projects in Ontario. To qualify for financial assistance, municipalities must play a central role in the implementation and ongoing development of recycling; applications will be supported only if there is adequate and reasonable commitment from markets for recovered materials; recycling systems must operate within the framework of a complete waste management system in which cost effectiveness is an important factor; multi-material projects are encouraged as much as possible; and the Ministry will share the costs of projects with the municipalities. The Ministry provides grants for up to 5 years per project to cover the net operating cost of a project up to a specified maximum percentage of eligible gross operating expenses. This manual provides guidelines for applying for such funding, including definitions of eligibility for operating and capital costs, the use of household bins, and guidelines for promotion and advertising, education, demonstration, and feasibility studies.

Coir dust reinforced recycled polypropylene composites; Compositos de polipropileno reciclado e po de coco

Energy Technology Data Exchange (ETDEWEB)

Santos, Bianca B. dos; Costa, Marysilvia F. da; Thire, Rossana M. da S.M., E-mail: bianca@metalmat.ufrj.br [Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ (Brazil)

2015-07-01

The environmental impacts caused by disposed plastics encourage the search for new alternatives. Recycling polymers leads to the degradation of their mechanical properties, which can be modified by the addition of fillers. In this paper, recycled polypropylene from plastic cups with 2%, 5% and 10% of coir dust were produced with and without the addition of additives. These composites were characterized by tensile tests, X-ray diffraction, differential scanning calorimetry and scanning electron microscopy on the fracture surface. It was verified the effectiveness of the addition of coir dust in improving the elasticity modulus of recycled polypropylene besides the effectiveness of additives used in promoting the adhesion of the powder to the matrix. However, higher levels of coir dust caused the appearance of air bubbles inside the material, which contributed to its embrittlement. The addition of coir dust promoted a decrease in the degree of polypropylene crystallinity. (author)

Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework

KAUST Repository

Alsaiari, Shahad K.

2017-12-22

CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days employing CRISPR/Cas9 machinery. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled co-delivery of intact Cas9 protein and sgRNA.

Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework

KAUST Repository

Alsaiari, Shahad K.; Patil, Sachin; Alyami, Mram Z.; Alamoudi, Kholod; Aleisa, Fajr A; Merzaban, Jasmeen; Li, Mo; Khashab, Niveen M.

2017-01-01

CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days employing CRISPR/Cas9 machinery. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled co-delivery of intact Cas9 protein and sgRNA.

Open-loop recycling: A LCA case study of PET bottle-to-fibre-recycling

NARCIS (Netherlands)

Shen, L.; Worrell, E.; Patel, M.K.

2010-01-01

This study assesses the environmental impact of polyethylene terephthalate (PET) bottle-to-fibre recycling using the methodology of life-cycle assessment (LCA). Four recycling cases, including mechanical recycling, semi-mechanical recycling, back-to-oligomer recycling and back-to-monomer recycling

Ligand-induced internalization of neurotensin in transfected COS-7 cells: differential intracellular trafficking of ligand and receptor.

Science.gov (United States)

Vandenbulcke, F; Nouel, D; Vincent, J P; Mazella, J; Beaudet, A

2000-09-01

The neuropeptide neurotensin (NT) is known to be internalized in a receptor-mediated fashion into its target cells. To gain insight into the mechanisms underlying this process, we monitored in parallel the migration of the NT1 neurotensin receptor subtype and a fluorescent analog of NT (fluo-NT) in COS-7 cells transfected with a tagged NT1 construct. Fluo-NT internalization was prevented by hypertonic sucrose, potassium depletion and cytosol acidification, demonstrating that it proceeded via clathrin-coated pits. Within 0-30 minutes, fluo-NT accumulated together with its receptor in Acridine Orange-positive, acidic organelles. These organelles concentrated transferrin and immunostained positively for rab 5A, therefore they were early endosomes. After 30-45 minutes, the ligand and its receptor no longer colocalized. Fluo-NT was first found in rab 7-positive late endosomes and later in a nonacidic juxtanuclear compartment identified as the Trans-Golgi Network (TGN) by virtue of its staining for syntaxin 6. This juxtanuclear compartment also stained positively for rab 7 and for the TGN/pericentriolar recycling endosome marker rab 11, suggesting that the ligand could have been recruited to the TGN from either late or recycling endosomes. By that time, internalized receptors were detected in Lamp-1-immunoreactive lysosomes. These results demonstrate that neurotensin/NT1 receptor complexes follow a recycling cycle that is unique among the G protein-coupled receptors studied to date, and provide the first evidence for the targeting of a nonendogenous protein from endosomes to the TGN.

Modeling the signaling endosome hypothesis: Why a drive to the nucleus is better than a (random walk

Directory of Open Access Journals (Sweden)

Howe Charles L

2005-10-01

Full Text Available Abstract Background Information transfer from the plasma membrane to the nucleus is a universal cell biological property. Such information is generally encoded in the form of post-translationally modified protein messengers. Textbook signaling models typically depend upon the diffusion of molecular signals from the site of initiation at the plasma membrane to the site of effector function within the nucleus. However, such models fail to consider several critical constraints placed upon diffusion by the cellular milieu, including the likelihood of signal termination by dephosphorylation. In contrast, signaling associated with retrogradely transported membrane-bounded organelles such as endosomes provides a dephosphorylation-resistant mechanism for the vectorial transmission of molecular signals. We explore the relative efficiencies of signal diffusion versus retrograde transport of signaling endosomes. Results Using large-scale Monte Carlo simulations of diffusing STAT-3 molecules coupled with probabilistic modeling of dephosphorylation kinetics we found that predicted theoretical measures of STAT-3 diffusion likely overestimate the effective range of this signal. Compared to the inherently nucleus-directed movement of retrogradely transported signaling endosomes, diffusion of STAT-3 becomes less efficient at information transfer in spatial domains greater than 200 nanometers from the plasma membrane. Conclusion Our model suggests that cells might utilize two distinct information transmission paradigms: 1 fast local signaling via diffusion over spatial domains on the order of less than 200 nanometers; 2 long-distance signaling via information packets associated with the cytoskeletal transport apparatus. Our model supports previous observations suggesting that the signaling endosome hypothesis is a subset of a more general hypothesis that the most efficient mechanism for intracellular signaling-at-a-distance involves the association of signaling

Vps33B is required for delivery of endocytosed cargo to lysosomes.

Science.gov (United States)

Galmes, Romain; ten Brink, Corlinda; Oorschot, Viola; Veenendaal, Tineke; Jonker, Caspar; van der Sluijs, Peter; Klumperman, Judith

2015-12-01

Lysosomes are the main degradative compartments of eukaryotic cells. The CORVET and HOPS tethering complexes are well known for their role in membrane fusion in the yeast endocytic pathway. Yeast Vps33p is part of both complexes, and has two mammalian homologues: Vps33A and Vps33B. Vps33B is required for recycling of apical proteins in polarized cells and a causative gene for ARC syndrome. Here, we investigate whether Vps33B is also required in the degradative pathway. By fluorescence and electron microscopy we show that Vps33B depletion in HeLa cells leads to significantly increased numbers of late endosomes that together with lysosomes accumulate in the perinuclear region. Degradation of endocytosed cargo is impaired in these cells. By electron microscopy we show that endocytosed BSA-gold reaches late endosomes, but is decreased in lysosomes. The increase in late endosome numbers and the lack of internalized cargo in lysosomes are indicative for a defect in late endosomal-lysosomal fusion events, which explains the observed decrease in cargo degradation. A corresponding phenotype was found after Vps33A knock down, which in addition also resulted in decreased lysosome numbers. We conclude that Vps33B, in addition to its role in endosomal recycling, is required for late endosomal-lysosomal fusion events. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

The effects of recycling loops in food waste management in Japan: based on the environmental and economic evaluation of food recycling.

Science.gov (United States)

Takata, Miki; Fukushima, Kazuyo; Kino-Kimata, Noriko; Nagao, Norio; Niwa, Chiaki; Toda, Tatsuki

2012-08-15

In Japan, a revised Food Recycling Law went into effect in 2007 to promote a "recycling loop" that requires food industries to purchase farm products that are grown using food waste-derived compost/animal feed. To realize and expand food recycling, it is necessary to evaluate how the recycling facilities work in the recycling loop. The purpose of this study is to assess the environmental and economic efficiency of the food recycling facilities that are involved in the recycling loop, which are also known as looped facilities. The global warming potential and running cost of five looped facilities were evaluated by LCA (life cycle assessment) and LCC (life cycle cost) approaches: machine integrated compost, windrow compost, liquid feed, dry feed, and bio-gasification. The LCA results showed low total GHG (greenhouse gas) emissions of -126 and -49 kg-CO(2)/t-waste, respectively, for dry feed and bio-gasification facilities, due to a high substitution effect. The LCC study showed a low running cost for composting facilities of -15,648 and -18,955 yen/t-waste, respectively, due to high revenue from the food waste collection. It was found that the mandatory reporting of food waste emitters to the government increased collection fees; however, the collection fee in animal feed facilities was relatively low because food waste was collected at a low price or nutritious food waste was purchased to produce quality feed. In the characterisation survey of various treatment methods, the composting facilities showed a relatively low environmental impact and a high economic efficiency. Animal feed facilities had a wide distribution of the total GHG emissions, depending on both the energy usage during the drying process and the substitution effect, which were related to the water content of the food waste and the number of recycled products. In comparison with incineration, the majority of the food recycling facilities showed low GHG emissions and economic effectiveness. This

The effects of recycling loops in food waste management in Japan: Based on the environmental and economic evaluation of food recycling

International Nuclear Information System (INIS)

Takata, Miki; Fukushima, Kazuyo; Kino-Kimata, Noriko; Nagao, Norio; Niwa, Chiaki; Toda, Tatsuki

2012-01-01

In Japan, a revised Food Recycling Law went into effect in 2007 to promote a “recycling loop” that requires food industries to purchase farm products that are grown using food waste-derived compost/animal feed. To realize and expand food recycling, it is necessary to evaluate how the recycling facilities work in the recycling loop. The purpose of this study is to assess the environmental and economic efficiency of the food recycling facilities that are involved in the recycling loop, which are also known as looped facilities. The global warming potential and running cost of five looped facilities were evaluated by LCA (life cycle assessment) and LCC (life cycle cost) approaches: machine integrated compost, windrow compost, liquid feed, dry feed, and bio-gasification. The LCA results showed low total GHG (greenhouse gas) emissions of − 126 and − 49 kg-CO 2 /t-waste, respectively, for dry feed and bio-gasification facilities, due to a high substitution effect. The LCC study showed a low running cost for composting facilities of − 15,648 and − 18,955 yen/t-waste, respectively, due to high revenue from the food waste collection. It was found that the mandatory reporting of food waste emitters to the government increased collection fees; however, the collection fee in animal feed facilities was relatively low because food waste was collected at a low price or nutritious food waste was purchased to produce quality feed. In the characterisation survey of various treatment methods, the composting facilities showed a relatively low environmental impact and a high economic efficiency. Animal feed facilities had a wide distribution of the total GHG emissions, depending on both the energy usage during the drying process and the substitution effect, which were related to the water content of the food waste and the number of recycled products. In comparison with incineration, the majority of the food recycling facilities showed low GHG emissions and economic

The effects of recycling loops in food waste management in Japan: Based on the environmental and economic evaluation of food recycling

Energy Technology Data Exchange (ETDEWEB)

Takata, Miki [Graduate School of Engineering, Soka University, Hachioji, Tokyo 192-8577 (Japan); Fukushima, Kazuyo [Watanabe Oyster Laboratory Co., Ltd, Hachioji, Tokyo 192-0154 (Japan); Kino-Kimata, Noriko [Graduate School of Engineering, Soka University, Hachioji, Tokyo 192-8577 (Japan); Nagao, Norio [Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor (Malaysia); Niwa, Chiaki [Graduate School of Engineering, Soka University, Hachioji, Tokyo 192-8577 (Japan); Toda, Tatsuki, E-mail: toda@soka.ac.jp [Graduate School of Engineering, Soka University, Hachioji, Tokyo 192-8577 (Japan)

2012-08-15

In Japan, a revised Food Recycling Law went into effect in 2007 to promote a 'recycling loop' that requires food industries to purchase farm products that are grown using food waste-derived compost/animal feed. To realize and expand food recycling, it is necessary to evaluate how the recycling facilities work in the recycling loop. The purpose of this study is to assess the environmental and economic efficiency of the food recycling facilities that are involved in the recycling loop, which are also known as looped facilities. The global warming potential and running cost of five looped facilities were evaluated by LCA (life cycle assessment) and LCC (life cycle cost) approaches: machine integrated compost, windrow compost, liquid feed, dry feed, and bio-gasification. The LCA results showed low total GHG (greenhouse gas) emissions of - 126 and - 49 kg-CO{sub 2}/t-waste, respectively, for dry feed and bio-gasification facilities, due to a high substitution effect. The LCC study showed a low running cost for composting facilities of - 15,648 and - 18,955 yen/t-waste, respectively, due to high revenue from the food waste collection. It was found that the mandatory reporting of food waste emitters to the government increased collection fees; however, the collection fee in animal feed facilities was relatively low because food waste was collected at a low price or nutritious food waste was purchased to produce quality feed. In the characterisation survey of various treatment methods, the composting facilities showed a relatively low environmental impact and a high economic efficiency. Animal feed facilities had a wide distribution of the total GHG emissions, depending on both the energy usage during the drying process and the substitution effect, which were related to the water content of the food waste and the number of recycled products. In comparison with incineration, the majority of the food recycling facilities showed low GHG emissions and economic

Behaviour of Recycled Coarse Aggregate Concrete: Age and Successive Recycling

Science.gov (United States)

Sahoo, Kirtikanta; Pathappilly, Robin Davis; Sarkar, Pradip

2016-06-01

Recycled Coarse Aggregate (RCA) concrete construction technique can be called as `green concrete', as it minimizes the environmental hazard of the concrete waste disposal. Indian standard recommends target mean compressive strength of the conventional concrete in terms of water cement ratio ( w/ c). The present work is an attempt to study the behaviour of RCA concrete from two samples of parent concrete having different age group with regard to the relationship of compressive strength with water cement ratios. Number of recycling may influence the mechanical properties of RCA concrete. The influence of age and successive recycling on the properties such as capillary water absorption, drying shrinkage strain, air content, flexural strength and tensile splitting strength of the RCA concrete are examined. The relationship between compressive strength at different w/ c ratios obtained experimentally is investigated for the two parameters such as age of parent concrete and successive recycling. The recycled concrete using older recycled aggregate shows poor quality. While the compressive strength reduces with successive recycling gradually, the capillary water absorption increases abruptly, which leads to the conclusion that further recycling may not be advisable.

Emergy analysis of the recycling options for construction and demolition waste.

Science.gov (United States)

Yuan, Fang; Shen, Li-yin; Li, Qi-ming

2011-12-01

Construction and demolition (C&D) waste is becoming a major contributor to environmental pollution. In Shanghai, China, the quantity of C&D waste is 2.11E+07 t/yr, which accounts for 45% of the total quantity of solid waste. There has been a growing promotion of recycling C&D waste as an effective way to solve this waste problem. However, the evaluation of the efficiency of recycling C&D waste as a potential source of resources is largely based on traditional economic analysis. The economic analysis emphasizes money instead of the harmony between economic benefit and environmental effects. There is a need for a new strategic approach to investigate the efficiency of recycling C&D waste to achieve the integration between economic, social and environmental effects. Emergy theory can be employed to analyze different recycling options for C&D waste. With reference to the Chinese construction industry, this paper demonstrates that the close-loop recycling option is better than the open-loop recycling option for C&D waste in terms of the integration of social, environmental and sustainable aspects. To evaluate different technology solutions for C&D waste recycling, the emergy theory and method is not limited to a cost-benefit balance but can include economic, social, environmental and sustainable effects. Copyright © 2011 Elsevier Ltd. All rights reserved.

Influence of recycled polystyrene beads on cement paste properties

Directory of Open Access Journals (Sweden)

Maaroufi Maroua

2018-01-01

Full Text Available In order to keep up with the requirements of sustainable development, there is a growing interest towards reducing the energy consumption in the construction and rehabilitation of buildings and the promotion of recycling waste in building materials. The use of recycled polystyrene beads in cement-based materials composition constitutes a solution to improve the insulation in buildings. This allows also limiting landfill by reusing the polystyrene waste. The aim of this study is to compare some properties and performances of a cement paste containing polystyrene beads to a reference paste designed with only the same cement. An experimental campaign was conducted and the obtained results showed that adding recycled polystyrene beads to a cement paste improves its hygro-thermal properties. Further studies are however necessary to better understand the real role of the polystyrene beads in the heat and mass transfers.

Delayed internalization and lack of recycling in a beta2-adrenergic receptor fused to the G protein alpha-subunit

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Floridi Aristide

2008-10-01

Full Text Available Abstract Background Chimeric proteins obtained by the fusion of a G protein-coupled receptor (GPCR sequence to the N-terminus of the G protein α-subunit have been extensively used to investigate several aspects of GPCR signalling. Although both the receptor and the G protein generally maintain a fully functional state in such polypeptides, original observations made using a chimera between the β2-adrenergic receptor (β2AR and Gαs indicated that the fusion to the α-subunit resulted in a marked reduction of receptor desensitization and down-regulation. To further investigate this phenomenon, we have compared the rates of internalization and recycling between wild-type and Gαs-fused β2AR. Results The rate of agonist-induced internalization, measured as the disappearance of cell surface immunofluorescence in HEK293 cells permanently expressing N-terminus tagged receptors, was reduced three-fold by receptor-G protein fusion. However, both fused and non-fused receptors translocated to the same endocytic compartment, as determined by dual-label confocal analysis of cells co-expressing both proteins and transferrin co-localization. Receptor recycling, determined as the reversion of surface immunofluorescence following the addition of antagonist to cells that were previously exposed to agonist, markedly differed between wild-type and fused receptors. While most of the internalized β2AR returned rapidly to the plasma membrane, β2AR-Gαs did not recycle, and the observed slow recovery for the fusion protein immunofluorescence was entirely accounted for by protein synthesis. Conclusion The covalent linkage between β2AR and Gαs does not appear to alter the initial endocytic translocation of the two proteins, although there is reduced efficiency. It does, however, completely disrupt the process of receptor and G protein recycling. We conclude that the physical separation between receptor and Gα is not necessary for the transit to early endosomes

The ArfGAP2/3 Glo3 and ergosterol collaborate in transport of a subset of cargoes

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Alejandro F. Estrada

2015-07-01

Full Text Available Proteins reach the plasma membrane through the secretory pathway in which the trans Golgi network (TGN acts as a sorting station. Transport from the TGN to the plasma membrane is maintained by a number of different pathways that act either directly or via the endosomal system. Here we show that a subset of cargoes depends on the ArfGAP2/3 Glo3 and ergosterol to maintain their proper localization at the plasma membrane. While interfering with neither ArfGAP2/3 activity nor ergosterol biosynthesis individually significantly altered plasma membrane localization of the tryptophan transporter Tat2, the general amino acid permease Gap1 and the v-SNARE Snc1, in a Δglo3 Δerg3 strain those proteins accumulated in internal endosomal structures. Export from the TGN to the plasma membrane and recycling from early endosomes appeared unaffected as the chitin synthase Chs3 that travels along these routes was localized normally. Our data indicate that a subset of proteins can reach the plasma membrane efficiently but after endocytosis becomes trapped in endosomal structures. Our study supports a role for ArfGAP2/3 in recycling from endosomes and in transport to the vacuole/lysosome.

The potential environmental gains from recycling waste plastics: simulation of transferring recycling and recovery technologies to Shenyang, China.

Science.gov (United States)

Chen, Xudong; Xi, Fengming; Geng, Yong; Fujita, Tsuyoshi

2011-01-01

With the increasing attention on developing a low-carbon economy, it is necessary to seek appropriate ways on reducing greenhouse gas (GHG) emissions through innovative municipal solid waste management (MSWM), such as urban symbiosis. However, quantitative assessments on the environmental benefits of urban symbiosis, especially in developing countries, are limited because only a limited number of planned synergistic activities have been successful and it is difficult to acquire detailed inventory data from private companies. This paper modifies and applies a two-step simulation system and used it to assess the potential environmental benefits, including the reduction of GHG emissions and saving of fossil fuels, by employing various Japanese plastics recycling/energy-recovery technologies in Shenyang, China. The results showed that among various recycling/energy-recovery technologies, the mechanical waste plastics recycling technology, which produces concrete formwork boards (NF boards), has the greatest potential in terms of reducing GHG emissions (1.66 kg CO(2)e/kg plastics), whereas the technology for the production of refuse plastic fuel (RPF) has the greatest potential on saving fossil fuel consumption (0.77 kg ce/kg-plastics). Additional benefits can be gained by applying combined technologies that cascade the utilization of waste plastics. Moreover, the development of clean energy in conjunction with the promotion of new waste plastics recycling programs could contribute to additional reductions in GHG emissions and fossil fuel consumption. Copyright © 2010 Elsevier Ltd. All rights reserved.

NEU3 Sialidase Protein Interactors in the Plasma Membrane and in the Endosomes*

Science.gov (United States)

Cirillo, Federica; Ghiroldi, Andrea; Fania, Chiara; Piccoli, Marco; Torretta, Enrica; Tettamanti, Guido; Gelfi, Cecilia; Anastasia, Luigi

2016-01-01

NEU3 sialidase has been shown to be a key player in many physio- and pathological processes, including cell differentiation, cellular response to hypoxic stress, and carcinogenesis. The enzyme, peculiarly localized on the outer leaflet of the plasma membrane, has been shown to be able to remove sialic acid residues from the gangliosides present on adjacent cells, thus creating cell to cell interactions. Nonetheless, herein we report that the enzyme localization is dynamically regulated between the plasma membrane and the endosomes, where a substantial amount of NEU3 is stored with low enzymatic activity. However, under opportune stimuli, NEU3 is shifted from the endosomes to the plasma membrane, where it greatly increases the sialidase activity. Finally, we found that NEU3 possesses also the ability to interact with specific proteins, many of which are different in each cell compartment. They were identified by mass spectrometry, and some selected ones were also confirmed by cross-immunoprecipitation with the enzyme, supporting NEU3 involvement in the cell stress response, protein folding, and intracellular trafficking. PMID:26987901

Effective photo-enhancement of cellular activity of fluorophore-octaarginine antisense PNA conjugates correlates with singlet oxygen formation, endosomal escape and chromophore lipophilicity

DEFF Research Database (Denmark)

Yarani, Reza; Shiraishi, Takehiko; Nielsen, Peter E.

2018-01-01

Photochemical internalization (PCI) is a cellular drug delivery method based on the generation of light-induced reactive oxygen species (ROS) causing damage to the endosomal membrane and thereby resulting in drug release to the cytoplasm. In our study a series of antisense fluorophore octaarginin...... indicate that efficient photodynamic endosomal escape is strongly dependent on the quantum yield for photochemical singlet oxygen formation, photostability as well as the lipophilicity of the chromophore....

Increased expression of endosomal members of toll-like receptor family abrogates wound healing in patients with type 2 diabetes mellitus.

Science.gov (United States)

Singh, Kanhaiya; Agrawal, Neeraj K; Gupta, Sanjeev K; Mohan, Gyanendra; Chaturvedi, Sunanda; Singh, Kiran

2016-10-01

The inflammatory phase of wound healing cascade is an important determinant of the fate of the wound. Acute inflammation is necessary to initiate proper wound healing, while chronic inflammation abrogates wound healing. Different endosomal members of toll-like receptor (TLR) family initiate inflammatory signalling via a range of different inflammatory mediators such as interferons, internal tissue damaged-associated molecular patterns (DAMPs) and hyperactive effector T cells. Sustained signalling of TLR9 and TLR7 contributes to chronic inflammation by activating the plasmacytoid dendritic cells. Diabetic wounds are also characterised by sustained inflammatory phase. The objective of this study was to analyse the differential expression of endosomal TLRs in human diabetic wounds compared with control wounds. We analysed the differential expression of TLR7 and TLR9 both at transcriptional and translational levels in wounds of 84 patients with type 2 diabetes mellitus (T2DM) and 6 control subjects without diabetes using quantitative real-time polymerase chain reaction (RT-PCR), western blot and immunohistochemistry. TLR7 and TLR9 were significantly up-regulated in wounds of the patients with T2DM compared with the controls and were dependent on the infection status of the diabetic wounds, and wounds with microbial infection exhibited lower expression levels of endosomal TLRs. Altered endosomal TLR expression in T2DM subjects might be associated with wound healing impairment. © 2015 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

End-of-life vehicle recycling and international cooperation between Japan, China and Korea: Present and future scenario analysis.

Science.gov (United States)

Che, Jia; Yu, Jeong-Soo; Kevin, Roy Serrona

2011-06-01

In the area of end-of-life vehicle (ELV) recycling, Japan passed the Automobile Recycling Law in January 2005, the first in Asia. Korea followed suit with the passage of the resource circulation method in 2009. China is expected make a new recycling law in 2011. In contribution to these initiatives, Tohoku University made a comparative analysis of ELV recycling laws, advance dismantling experiments and scenario analysis to promote international cooperation. This is envisioned to introduce ELV recycling system in Japan, China and Korea and in developing countries as well. Copyright © 2011 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Feasibility Assessments of the Use of Recycled Fibers in Nonwoven Fabrics

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Jia-Horng Lin

2017-01-01

Full Text Available Environmental protection has become an increasing concern, which makes recycling and reclaiming highly important. In addition to governmental campaigns and promotion, enterprises should examine each perspective thoroughly in order to prevent excessive resource consumption. In this study, recycled materials, including recycled far-infrared polyester (FPET fiber, three-dimensional crimped hollow flame-retarding (TPET fiber, and low-melting-point polyester (LPET fiber, are used to form nonwoven fabrics. The influence of different amounts of FPET fiber, 0–80 wt %, on the properties of nonwoven fabrics was examined. The sheath of LPET fibers can be melted as a result of hot pressing, which provides cohesion between fibers that mechanically improves the nonwoven fabrics. The tensile strength, tearing strength, air permeability, and far infrared (FIR emissivity of the nonwoven fabrics were examined, thereby determining the optimal parameters. The test results show that the thermally treated nonwoven fabrics have better mechanical properties and FIR emissivity, compared to those of non-thermally treated nonwoven fabrics. Moreover, more FPET fibers cause the mechanical properties along the cross machine direction (CD to decrease by 9% and that along the machine direction (MD to decrease by 5%. In particular, all the thermally treated samples exhibit a FIR emissivity of 0.8, which is health-promoting.

Rethink, Rework, Recycle.

Science.gov (United States)

Wrhen, Linda; DiSpezio, Michael A.

1991-01-01

Information about the recycling and reuse of plastics, aluminum, steel, glass, and newspapers is presented. The phases of recycling are described. An activity that allows students to separate recyclable materials is included. The objectives, a list of needed materials, and procedure are provided. (KR)

[Research on resources chemistry of Chinese medicinal materials and resources recycling utilization ways and goals and tasks].

Science.gov (United States)

Duan, Jin-ao; Su, Shu-lan; Guo, Sheng; Jiang, Shu; Liu, Pei; Yan, Hui; Qian, Da-wei; Zhu, Hua-xu; Tang, Yu-ping; Wu, Qi-nan

2015-09-01

The objects of research on the resources chemistry of Chinese medicinal materials (RCCMM) are promotion of efficient production, rational utilization and improving quality of CMM and natural products. The development of TCM cause depends on the efficient utilization and sustainable development of CMM, hinges on the technologies and methods for using and discovering medicinal biological resources, stand or fall on the extension of industy chains, detailed utilizaion of resource chemical components by multi-way, multi-level. All of these may help to the recycling utilization and sound development of RCMM. In this article, five respects were discussed to the RCCMM researches and resources recycling utilization ways and goals and tasks. First, based on the principle of resource scarcity, discovering or replacing CMM resources, protecting the rare or endangered species or resources. Second, based on the multifunctionality of CMM, realizing the value-added and value compensation, and promoting the utilization efficiency through systermatic and detailed exploitation and utilization. Third, based on the resource conservation and environment-friendly, reducing raw material consumption, lowering cost, promoting recycling utilization and elevating utilization efficiency. Fourth, based on the stratege of turning harm into good, using the invasive alien biological resources by multi-ways and enriching the medicial resources. Fifth, based on the method of structure modification of chemical components, exploring and enhancing the utility value of resouces chemical substances. These data should provide references and attention for improving the utilization efficiency, promoting the development of recycling economy, and changing the mode of economic growth of agriculture and industry of CMM fundamentally.

Research on Recycling and Utilization of Solid Waste in Civil Airport

Science.gov (United States)

Li, Bo; Zhang, Wen; Wang, Jianping; Yi, Wei

2018-05-01

The aviation industry is embracing unprecedented prosperity together with the economic development. Building green airports resource-saving, environment-friendly and sustainable has become the inevitability of the times. The operation of airport will generate the large amount of waste every day, which certainly exposes airports and surrounding regions to waste disposal and ecological environment pressure. Waste disposal directly affects the surrounding environment of airports, which can be effectively mitigated by disposing waste into resources, i.e., sorting and recycling them into renewable materials. The development of green airport can also be promoted in this process. The article elaborates on the current methods of waste disposal adopted by airports. According to the principle of waste reduction, harmlessness, and resource recycling, a set of solid waste recycling and utilization methods suitable for airports are proposed, which can reduce the costs of waste transported to other places and landfilled. Various environmental pollution caused by landfill and other disposal methods can also be contained effectively. At the same time, resources can be fully recycled, converting waste into useful resources in an efficient and environmental-friendly way.

Recycling of electronic scrap

DEFF Research Database (Denmark)

Legarth, Jens Brøbech

This Ph.D. thesis deals with the growingly important field of electronics recycling with special attention to the problem of printed circuit board recycling. A literature survey of contemporary electronics recycling and printed circuit board recycling is presented.Further, an analysis of the role...

Novel ethyl methanesulfonate (EMS-induced null alleles of the Drosophila homolog of LRRK2 reveal a crucial role in endolysosomal functions and autophagy in vivo

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Mark W. Dodson

2014-12-01

Full Text Available Mutations in LRRK2 cause a dominantly inherited form of Parkinson’s disease (PD and are the most common known genetic determinant of PD. Inhibitor-based therapies targeting LRRK2 have emerged as a key therapeutic strategy in PD; thus, understanding the consequences of inhibiting the normal cellular functions of this protein is vital. Despite much interest, the physiological functions of LRRK2 remain unclear. Several recent studies have linked the toxicity caused by overexpression of pathogenic mutant forms of LRRK2 to defects in the endolysosomal and autophagy pathways, raising the question of whether endogenous LRRK2 might play a role in these processes. Here, we report the characterization of multiple novel ethyl methanesulfonate (EMS-induced nonsense alleles in the Drosophila LRRK2 homolog, lrrk. Using these alleles, we show that lrrk loss-of-function causes striking defects in the endolysosomal and autophagy pathways, including the accumulation of markedly enlarged lysosomes that are laden with undigested contents, consistent with a defect in lysosomal degradation. lrrk loss-of-function also results in the accumulation of autophagosomes, as well as the presence of enlarged early endosomes laden with mono-ubiquitylated cargo proteins, suggesting an additional defect in lysosomal substrate delivery. Interestingly, the lysosomal abnormalities in these lrrk mutants can be suppressed by a constitutively active form of the small GTPase rab9, which promotes retromer-dependent recycling from late endosomes to the Golgi. Collectively, our data provides compelling evidence of a vital role for lrrk in lysosomal function and endolysosomal membrane transport in vivo, and suggests a link between lrrk and retromer-mediated endosomal recycling.

Novel ethyl methanesulfonate (EMS)-induced null alleles of the Drosophila homolog of LRRK2 reveal a crucial role in endolysosomal functions and autophagy in vivo.

Science.gov (United States)

Dodson, Mark W; Leung, Lok K; Lone, Mohiddin; Lizzio, Michael A; Guo, Ming

2014-12-01

Mutations in LRRK2 cause a dominantly inherited form of Parkinson's disease (PD) and are the most common known genetic determinant of PD. Inhibitor-based therapies targeting LRRK2 have emerged as a key therapeutic strategy in PD; thus, understanding the consequences of inhibiting the normal cellular functions of this protein is vital. Despite much interest, the physiological functions of LRRK2 remain unclear. Several recent studies have linked the toxicity caused by overexpression of pathogenic mutant forms of LRRK2 to defects in the endolysosomal and autophagy pathways, raising the question of whether endogenous LRRK2 might play a role in these processes. Here, we report the characterization of multiple novel ethyl methanesulfonate (EMS)-induced nonsense alleles in the Drosophila LRRK2 homolog, lrrk. Using these alleles, we show that lrrk loss-of-function causes striking defects in the endolysosomal and autophagy pathways, including the accumulation of markedly enlarged lysosomes that are laden with undigested contents, consistent with a defect in lysosomal degradation. lrrk loss-of-function also results in the accumulation of autophagosomes, as well as the presence of enlarged early endosomes laden with mono-ubiquitylated cargo proteins, suggesting an additional defect in lysosomal substrate delivery. Interestingly, the lysosomal abnormalities in these lrrk mutants can be suppressed by a constitutively active form of the small GTPase rab9, which promotes retromer-dependent recycling from late endosomes to the Golgi. Collectively, our data provides compelling evidence of a vital role for lrrk in lysosomal function and endolysosomal membrane transport in vivo, and suggests a link between lrrk and retromer-mediated endosomal recycling. © 2014. Published by The Company of Biologists Ltd.

Kinetic imaging of NPC1L1 and sterol trafficking between plasma membrane and recycling endosomes in hepatoma cells

DEFF Research Database (Denmark)

Hartwig Petersen, Nicole; Færgeman, Nils J; Yu, Liqing

2008-01-01

fluorescent protein (NPC1L1-EGFP) and cholesterol analogues in hepatoma cells. At steady state about 42% of NPC1L1 resided in the transferrin (Tf) positive, sterol enriched endocytic recycling compartment (ERC), while time-lapse microscopy demonstrated NPC1L1 traffic between plasma membrane and ERC...... the ERC to the plasma membrane. NPC1L1-EGFP facilitated transport of fluorescent sterols from the plasma membrane to the ERC. Insulin induced translocation of vesicles containing NPC1L1 and fluorescent sterol from the ERC to the cell membrane. Upon polarization of hepatoma cells NPC1L1 resided almost...... exclusively in the canalicular membrane, where the protein is highly mobile. Our study demonstrates dynamic trafficking of NPC1L1 between cell surface and intracellular compartments and suggests that this transport is involved in NPC1L1 mediated cellular sterol uptake....

Mutational analysis of the yeast TRAPP subunit Trs20p identifies roles in endocytic recycling and sporulation.

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Hichem Mahfouz

Full Text Available Trs20p is a subunit of the evolutionarily conserved TRAPP (TRAnsport Protein Particle complex that mediates various aspects of membrane trafficking. Three TRAPP complexes have been identified in yeast with roles in ER-to-Golgi trafficking, post-Golgi and endosomal-to-Golgi transport and in autophagy. The role of Trs20p, which is essential for viability and a component of all three complexes, and how it might function within each TRAPP complex, has not been clarified to date. To begin to address the role of Trs20p we generated different mutants by random mutagenesis but, surprisingly, no defects were observed in diverse anterograde transport pathways or general secretion in Trs20 temperature-sensitive mutants. Instead, mutation of Trs20 led to defects in endocytic recycling and a block in sporulation/meiosis. The phenotypes of different mutants appear to be separable suggesting that the mutations affect the function of Trs20 in different TRAPP complexes.

Caveolin targeting to late endosome/lysosomal membranes is induced by perturbations of lysosomal pH and cholesterol content

Science.gov (United States)

Mundy, Dorothy I.; Li, Wei Ping; Luby-Phelps, Katherine; Anderson, Richard G. W.

2012-01-01

Caveolin-1 is an integral membrane protein of plasma membrane caveolae. Here we report that caveolin-1 collects at the cytosolic surface of lysosomal membranes when cells are serum starved. This is due to an elevation of the intralysosomal pH, since ionophores and proton pump inhibitors that dissipate the lysosomal pH gradient also trapped caveolin-1 on late endosome/lysosomes. Accumulation is both saturable and reversible. At least a portion of the caveolin-1 goes to the plasma membrane upon reversal. Several studies suggest that caveolin-1 is involved in cholesterol transport within the cell. Strikingly, we find that blocking cholesterol export from lysosomes with progesterone or U18666A or treating cells with low concentrations of cyclodextrin also caused caveolin-1 to accumulate on late endosome/lysosomal membranes. Under these conditions, however, live-cell imaging shows cavicles actively docking with lysosomes, suggesting that these structures might be involved in delivering caveolin-1. Targeting of caveolin-1 to late endosome/lysosomes is not observed normally, and the degradation rate of caveolin-1 is not altered by any of these conditions, indicating that caveolin-1 accumulation is not a consequence of blocked degradation. We conclude that caveolin-1 normally traffics to and from the cytoplasmic surface of lysosomes during intracellular cholesterol trafficking. PMID:22238363

Modelling Recycling Targets

DEFF Research Database (Denmark)

hill, amanda; Leinikka Dall, Ole; Andersen, Frits Møller

2014-01-01

% for household waste, and sets an ambitious goal of a 50% recycling rate by 2020. This study integrates the recycling target into the FRIDA model to project how much waste and from which streams should be diverted from incineration to recycling in order to achieve the target. Furthermore, it discusses how...

On the effectiveness of a license scheme for E-waste recycling: The challenge of China and India

International Nuclear Information System (INIS)

Shinkuma, Takayoshi; Managi, Shunsuke

2010-01-01

It is well known that China and India have been recycling centers of WEEE, especially printed circuit boards, and that serious environmental pollution in these countries has been generated by improper recycling methods. After the governments of China and India banned improper recycling by the informal sector, improper recycling activities spread to other places. Then, these governments changed their policies to one of promoting proper recycling by introducing a scheme, under which E-waste recycling requires a license issued by the government. In this paper, the effectiveness of that license scheme is examined by means of an economic model. It can be shown that the license scheme can work effectively only if disposers of E-waste have a responsibility to sell E-waste to license holders. Our results run counter to the idea that international E-waste trade should be banned and provide an alternative solution to the problem.

Recycling production designs: the value of coordination and flexibility in aluminum recycling operations

Science.gov (United States)

Brommer, Tracey H.

The growing motivation for aluminum recycling has prompted interest in recycling alternative and more challenging secondary materials. The nature of these alternative secondary materials necessitates the development of an intermediate recycling facility that can reprocess the secondary materials into a liquid product Two downstream aluminum remelters will incorporate the liquid products into their aluminum alloy production schedules. Energy and environmental benefits result from delivering the products as liquid but coordination challenges persist because of the energy cost to maintain the liquid. Further coordination challenges result from the necessity to establish a long term recycling production plan in the presence of long term downstream aluminum remelter production uncertainty and inherent variation in the daily order schedule of the downstream aluminum remelters. In this context a fundamental question arises, considering the metallurgical complexities of dross reprocessing, what is the value of operating a coordinated set of by-product reprocessing plants and remelting cast houses? A methodology is presented to calculate the optimal recycling center production parameters including 1) the number of recycled products, 2) the volume of recycled products, 3) allocation of recycled materials across recycled products, 4) allocation of recycled products across finished alloys, 4) the level of flexibility for the recycling center to operate. The methods implemented include, 1) an optimization model to describe the long term operations of the recycling center, 2) an uncertainty simulation tool, 3) a simulation optimization method, 4) a dynamic simulation tool with four embedded daily production optimization models of varying degrees of flexibility. This methodology is used to quantify the performance of several recycling center production designs of varying levels of coordination and flexibility. This analysis allowed the identification of the optimal recycling

Strategies for the Activation and Release of the Membranolytic Peptide Melittin from Liposomes Using Endosomal pH as a Trigger.

Science.gov (United States)

Oude Blenke, E; Sleszynska, M; Evers, M J W; Storm, G; Martin, N I; Mastrobattista, E

2017-02-15

Endosomolytic peptides are often coupled to drug delivery systems to enhance endosomal escape, which is crucial for the delivery of macromolecular drugs that are vulnerable to degradation in the endolysosomal pathway. Melittin is a 26 amino acid peptide derived from bee venom that has a very high membranolytic activity. However, such lytic peptides also impose a significant safety risk when applied in vivo as they often have similar activity against red blood cells and other nontarget cell membranes. Our aim is to control the membrane-disrupting capacity of these peptides in time and space by physically constraining them to a nanocarrier surface in such a way that they only become activated when delivered inside acidic endosomes. To this end, a variety of chemical approaches for the coupling of lytic peptides to liposomes via functionalized PEG-lipids were explored, including maleimide-thiol chemistry, click-chemistry, and aldehyde-hydrazide chemistry. The latter enables reversible conjugation via a hydrazone bond, allowing for release of the peptide under endosomal conditions. By carefully choosing the conjugation site and by using a pH activated analog of the melittin peptide, lytic activity toward a model membrane is completely inhibited at physiological pH. At endosomal pH the activity is restored by hydrolysis of the acid-labile hydrazone bond, releasing the peptide in its most active, free form. Furthermore, using an analogue containing a nonhydrolyzable bond as a control, it was shown that the activity observed can be completely attributed to release of the peptide, validating dynamic covalent conjugation as a suitable strategy to maintain safety during circulation.

Structural Studies of Adeno-Associated Virus Serotype 8 Capsid Transitions Associated with Endosomal Trafficking

Energy Technology Data Exchange (ETDEWEB)

Nam, Hyun-Joo; Gurda, Brittney L.; McKenna, Robert; Potter, Mark; Byrne, Barry; Salganik, Maxim; Muzyczka, Nicholas; Agbandje-McKenna, Mavis (Florida)

2012-09-17

The single-stranded DNA (ssDNA) parvoviruses enter host cells through receptor-mediated endocytosis, and infection depends on processing in the early to late endosome as well as in the lysosome prior to nuclear entry for replication. However, the mechanisms of capsid endosomal processing, including the effects of low pH, are poorly understood. To gain insight into the structural transitions required for this essential step in infection, the crystal structures of empty and green fluorescent protein (GFP) gene-packaged adeno-associated virus serotype 8 (AAV8) have been determined at pH values of 6.0, 5.5, and 4.0 and then at pH 7.5 after incubation at pH 4.0, mimicking the conditions encountered during endocytic trafficking. While the capsid viral protein (VP) topologies of all the structures were similar, significant amino acid side chain conformational rearrangements were observed on (i) the interior surface of the capsid under the icosahedral 3-fold axis near ordered nucleic acid density that was lost concomitant with the conformational change as pH was reduced and (ii) the exterior capsid surface close to the icosahedral 2-fold depression. The 3-fold change is consistent with DNA release from an ordering interaction on the inside surface of the capsid at low pH values and suggests transitions that likely trigger the capsid for genome uncoating. The surface change results in disruption of VP-VP interface interactions and a decrease in buried surface area between VP monomers. This disruption points to capsid destabilization which may (i) release VP1 amino acids for its phospholipase A2 function for endosomal escape and nuclear localization signals for nuclear targeting and (ii) trigger genome uncoating.

Concrete produced with recycled aggregates

Directory of Open Access Journals (Sweden)

J. J. L. Tenório

Full Text Available This paper presents the analysis of the mechanical and durable properties of recycled aggregate concrete (RAC for using in concrete. The porosity of recycled coarse aggregates is known to influence the fresh and hardened concrete properties and these properties are related to the specific mass of the recycled coarse aggregates, which directly influences the mechanical properties of the concrete. The recycled aggregates were obtained from construction and demolition wastes (CDW, which were divided into recycled sand (fine and coarse aggregates. Besides this, a recycled coarse aggregate of a specific mass with a greater density was obtained by mixing the recycled aggregates of the CDW with the recycled aggregates of concrete wastes (CW. The concrete was produced in laboratory by combining three water-cement ratios, the ratios were used in agreement with NBR 6118 for structural concretes, with each recycled coarse aggregates and recycled sand or river sand, and the reference concrete was produced with natural aggregates. It was observed that recycled aggregates can be used in concrete with properties for structural concrete. In general, the use of recycled coarse aggregate in combination with recycled sand did not provide good results; but when the less porous was used, or the recycled coarse aggregate of a specific mass with a greater density, the properties of the concrete showed better results. Some RAC reached bigger strengths than the reference concrete.

Tetraspanin CD63 Bridges Autophagic and Endosomal Processes To Regulate Exosomal Secretion and Intracellular Signaling of Epstein-Barr Virus LMP1

Science.gov (United States)

Hurwitz, Stephanie N; Cheerathodi, Mujeeb R; Nkosi, Dingani; York, Sara B; Meckes, David G

2018-03-01

The tetraspanin protein CD63 has been recently described as a key factor in extracellular vesicle (EV) production and endosomal cargo sorting. In the context of Epstein-Barr virus (EBV) infection, CD63 is required for the efficient packaging of the major viral oncoprotein latent membrane protein 1 (LMP1) into exosomes and other EV populations and acts as a negative regulator of LMP1 intracellular signaling. Accumulating evidence has also pointed to intersections of the endosomal and autophagy pathways in maintaining cellular secretory processes and as sites for viral assembly and replication. Indeed, LMP1 can activate the mammalian target of rapamycin (mTOR) pathway to suppress host cell autophagy and facilitate cell growth and proliferation. Despite the growing recognition of cross talk between endosomes and autophagosomes and its relevance to viral infection, little is understood about the molecular mechanisms governing endosomal and autophagy convergence. Here, we demonstrate that CD63-dependent vesicle protein secretion directly opposes intracellular signaling activation downstream of LMP1, including mTOR-associated proteins. Conversely, disruption of normal autolysosomal processes increases LMP1 secretion and dampens signal transduction by the viral protein. Increases in mTOR activation following CD63 knockout are coincident with the development of serum-dependent autophagic vacuoles that are acidified in the presence of high LMP1 levels. Altogether, these findings suggest a key role of CD63 in regulating the interactions between endosomal and autophagy processes and limiting cellular signaling activity in both noninfected and virally infected cells. IMPORTANCE The close connection between extracellular vesicles and viruses is becoming rapidly and more widely appreciated. EBV, a human gamma herpesvirus that contributes to the progression of a multitude of lymphomas and carcinomas in immunocompromised or genetically susceptible populations, packages its major

Green Science: Revisiting Recycling

Science.gov (United States)

Palliser, Janna

2011-01-01

Recycling has been around for a long time--people have reused materials and refashioned them into needed items for thousands of years. More recently, war efforts encouraged conservation and reuse of materials, and in the 1970s recycling got its official start when recycling centers were created. Now, curbside recycling programs and recycling…

A long-term study on the effect of magnetite supplementation in continuous anaerobic digestion of dairy effluent - Magnetic separation and recycling of magnetite.

Science.gov (United States)

Baek, Gahyun; Jung, Heejung; Kim, Jaai; Lee, Changsoo

2017-10-01

Promotion of direct interspecies electron transfer (DIET) between exoelectrogenic bacteria and electron-utilizing methanogens has recently been discussed as a new method for enhanced biomethanation. This study evaluated the effect of magnetite-promoted DIET in continuous anaerobic digestion of dairy effluent and tested the magnetic separation and recycling of magnetite to avoid continuous magnetite addition. The applied magnetite recycling method effectively supported enhanced DIET activity and biomethanation performance over a long period (>250days) without adding extra magnetite. DIET via magnetite particles as electrical conduits was likely the main mechanism for the enhanced biomethanation. Magnetite formed complex aggregate structures with microbes, and magnetite recycling also helped retain more biomass in the process. Methanosaeta was likely the major methanogen group responsible for DIET-based methanogenesis, in association with Proteobacteria and Chloroflexi populations as syntrophic partners. The recycling approach proved robust and effective, highlighting the potential of magnetite recycling for high-rate biomethanation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nuclear fuel cycle waste recycling technology deverlopment - Radioactive metal waste recycling technology development

International Nuclear Information System (INIS)

Oh, Won Zin; Moon, Jei Kwon; Jung, Chong Hun; Park, Sang Yoon

1998-08-01

With relation to recycling of the radioactive metal wastes which are generated during operation and decommissioning of nuclear facilities, the following were described in this report. 1. Analysis of the state of the art on the radioactive metal waste recycling technologies. 2. Economical assessment on the radioactive metal waste recycling. 3. Process development for radioactive metal waste recycling, A. Decontamination technologies for radioactive metal waste recycling. B. Decontamination waste treatment technologies, C. Residual radioactivity evaluation technologies. (author). 238 refs., 60 tabs., 79 figs

Endocytic recycling via the TGN underlies the polarized hyphal mode of life.

Science.gov (United States)

Hernández-González, Miguel; Bravo-Plaza, Ignacio; Pinar, Mario; de Los Ríos, Vivian; Arst, Herbert N; Peñalva, Miguel A

2018-04-01

Intracellular traffic in Aspergillus nidulans hyphae must cope with the challenges that the high rates of apical extension (1μm/min) and the long intracellular distances (>100 μm) impose. Understanding the ways in which the hyphal tip cell coordinates traffic to meet these challenges is of basic importance, but is also of considerable applied interest, as fungal invasiveness of animals and plants depends critically upon maintaining these high rates of growth. Rapid apical extension requires localization of cell-wall-modifying enzymes to hyphal tips. By combining genetic blocks in different trafficking steps with multidimensional epifluorescence microscopy and quantitative image analyses we demonstrate that polarization of the essential chitin-synthase ChsB occurs by indirect endocytic recycling, involving delivery/exocytosis to apices followed by internalization by the sub-apical endocytic collar of actin patches and subsequent trafficking to TGN cisternae, where it accumulates for ~1 min before being re-delivered to the apex by a RAB11/TRAPPII-dependent pathway. Accordingly, ChsB is stranded at the TGN by Sec7 inactivation but re-polarizes to the apical dome if the block is bypassed by a mutation in geaAgea1 that restores growth in the absence of Sec7. That polarization is independent of RAB5, that ChsB predominates at apex-proximal cisternae, and that upon dynein impairment ChsB is stalled at the tips in an aggregated endosome indicate that endocytosed ChsB traffics to the TGN via sorting endosomes functionally located upstream of the RAB5 domain and that this step requires dynein-mediated basipetal transport. It also requires RAB6 and its effector GARP (Vps51/Vps52/Vps53/Vps54), whose composition we determined by MS/MS following affinity chromatography purification. Ablation of any GARP component diverts ChsB to vacuoles and impairs growth and morphology markedly, emphasizing the important physiological role played by this pathway that, we propose, is central

An Empirical Test of an Expanded Version of the Theory of Planned Behavior in Predicting Recycling Behavior on Campus

Science.gov (United States)

Largo-Wight, Erin; Bian, Hui; Lange, Lori

2012-01-01

Background: The study and promotion of environmental health behaviors, such as recycling, is an emerging focus in public health. Purpose: This study was designed to examine the determinants of recycling intention on a college campus. Methods: Undergraduate students (N=189) completed a 35-item web-based survey past findings and an expanded version…

Syndecan-4 Phosphorylation Is a Control Point for Integrin Recycling

Science.gov (United States)

Morgan, Mark R.; Hamidi, Hellyeh; Bass, Mark D.; Warwood, Stacey; Ballestrem, Christoph; Humphries, Martin J.

2013-01-01

Summary Precise spatiotemporal coordination of integrin adhesion complex dynamics is essential for efficient cell migration. For cells adherent to fibronectin, differential engagement of α5β1 and αVβ3 integrins is used to elicit changes in adhesion complex stability, mechanosensation, matrix assembly, and migration, but the mechanisms responsible for receptor regulation have remained largely obscure. We identify phosphorylation of the membrane-intercalated proteoglycan syndecan-4 as an essential switch controlling integrin recycling. Src phosphorylates syndecan-4 and, by driving syntenin binding, leads to suppression of Arf6 activity and recycling of αVβ3 to the plasma membrane at the expense of α5β1. The resultant elevation in αVβ3 engagement promotes stabilization of focal adhesions. Conversely, abrogation of syndecan-4 phosphorylation drives surface expression of α5β1, destabilizes adhesion complexes, and disrupts cell migration. These data identify the dynamic spatiotemporal regulation of Src-mediated syndecan-4 phosphorylation as an essential switch controlling integrin trafficking and adhesion dynamics to promote efficient cell migration. PMID:23453597

Usage of Recycled Pet

Directory of Open Access Journals (Sweden)

A. Ebru Tayyar

2010-01-01

Full Text Available The increasing industrialization, urbanization and the technological development have caused to increase depletion of the natural resources and environmental pollution's problem. Especially, for the countries which have not enough space recycling of the waste eliminating waste on regular basis or decreasing the amount and volume of waste have provided the important advantages. There are lots of studies and projects to develop both protect resources and prevent environmental pollution. PET bottles are commonly used in beverage industry and can be reused after physical and chemical recycling processes. Usage areas of recycled PET have been developed rapidly. Although recycled PET is used in plastic industry, composite industry also provides usage alternatives of recycled PET. Textile is a suitable sector for recycling of some plastics made of polymers too. In this study, the recycling technologies and applications of waste PET bottles have been investigated and scientific works in this area have been summarized.

The late endosomal HOPS complex anchors active G-protein signaling essential for pathogenesis in magnaporthe oryzae.

Directory of Open Access Journals (Sweden)

Ravikrishna Ramanujam

Full Text Available In Magnaporthe oryzae, the causal ascomycete of the devastating rice blast disease, the conidial germ tube tip must sense and respond to a wide array of requisite cues from the host in order to switch from polarized to isotropic growth, ultimately forming the dome-shaped infection cell known as the appressorium. Although the role for G-protein mediated Cyclic AMP signaling in appressorium formation was first identified almost two decades ago, little is known about the spatio-temporal dynamics of the cascade and how the signal is transmitted through the intracellular network during cell growth and morphogenesis. In this study, we demonstrate that the late endosomal compartments, comprising of a PI3P-rich (Phosphatidylinositol 3-phosphate highly dynamic tubulo-vesicular network, scaffold active MagA/GαS, Rgs1 (a GAP for MagA, Adenylate cyclase and Pth11 (a non-canonical GPCR in the likely absence of AKAP-like anchors during early pathogenic development in M. oryzae. Loss of HOPS component Vps39 and consequently the late endosomal function caused a disruption of adenylate cyclase localization, cAMP signaling and appressorium formation. Remarkably, exogenous cAMP rescued the appressorium formation defects associated with VPS39 deletion in M. oryzae. We propose that sequestration of key G-protein signaling components on dynamic late endosomes and/or endolysosomes, provides an effective molecular means to compartmentalize and control the spatio-temporal activation and rapid downregulation (likely via vacuolar degradation of cAMP signaling amidst changing cellular geometry during pathogenic development in M. oryzae.

Algal recycling enhances algal productivity and settleability in Pediastrum boryanum pure cultures.

Science.gov (United States)

Park, Jason B K; Craggs, Rupert J; Shilton, Andy N

2015-12-15

Recycling a portion of gravity harvested algae (i.e. algae and associated bacteria biomass) has been shown to improve both algal biomass productivity and harvest efficiency by maintaining the dominance of a rapidly-settleable colonial alga, Pediastrum boryanum in both pilot-scale wastewater treatment High Rate Algal Ponds (HRAP) and outdoor mesocosms. While algal recycling did not change the relative proportions of algae and bacteria in the HRAP culture, the contribution of the wastewater bacteria to the improved algal biomass productivity and settleability with the recycling was not certain and still required investigation. P. boryanum was therefore isolated from the HRAP and grown in pure culture on synthetic wastewater growth media under laboratory conditions. The influence of recycling on the productivity and settleability of the pure P. boryanum culture was then determined without wastewater bacteria present. Six 1 L P. boryanum cultures were grown over 30 days in a laboratory growth chamber simulating New Zealand summer conditions either with (Pr) or without (Pc) recycling of 10% of gravity harvested algae. The cultures with recycling (Pr) had higher algal productivity than the controls (Pc) when the cultures were operated at both 4 and 3 d hydraulic retention times by 11% and 38% respectively. Furthermore, algal recycling also improved 1 h settleability from ∼60% to ∼85% by increasing the average P. boryanum colony size due to the extended mean cell residence time and promoted formation of large algal bio-flocs (>500 μm diameter). These results demonstrate that the presence of wastewater bacteria was not necessary to improve algal productivity and settleability with algal recycling. Copyright © 2015 Elsevier Ltd. All rights reserved.

Assessing changes on poly(ethylene terephthalate) properties after recycling: Mechanical recycling in laboratory versus postconsumer recycled material

Energy Technology Data Exchange (ETDEWEB)

López, María del Mar Castro, E-mail: quimcl02@udc.es [Grupo de Polímeros, Centro de Investigacións Tecnológicas (CIT), Departamento de Física, Escuela Universitaria Politécnica, Universidade de A Coruña, Campus de Ferrol, 15403 Ferrol (Spain); Ares Pernas, Ana Isabel, E-mail: aares@udc.es [Grupo de Polímeros, Centro de Investigacións Tecnológicas (CIT), Departamento de Física, Escuela Universitaria Politécnica, Universidade de A Coruña, Campus de Ferrol, 15403 Ferrol (Spain); Abad López, Ma José, E-mail: mjabad@udc.es [Grupo de Polímeros, Centro de Investigacións Tecnológicas (CIT), Departamento de Física, Escuela Universitaria Politécnica, Universidade de A Coruña, Campus de Ferrol, 15403 Ferrol (Spain); and others

2014-10-15

Keeping rheological, mechanical and thermal properties of virgin poly(ethylene terephthalate), PET, is necessary to assure the quality of second-market applications. A comparative study of these properties has been undertaken in virgin, mechanical recycled and commercial recycled PET samples. Viscoelastic characterization was carried out by rheological measurements. Mechanical properties were estimated by tensile and Charpy impact strength tests. Thermal properties and crystallinity were evaluated by differential scanning calorimetry and a deconvolution procedure was applied to study the population of the different crystals. Molecular conformational changes related to crystallinity values were studied by FTIR spectroscopy. Variations in average molecular weight were predicted from rheology. Besides, the presence-absence of linear and cyclic oligomeric species was measured by mass spectrometry techniques, as MALDI-TOF. Mechanical recycled PET undergoes a significant decline in rheological, mechanical and thermal properties upon increasing the number of reprocessing steps. This is due to the cleavage of the ester bonds with reduction in molar mass and raise in cyclic oligomeric species, in particular [GT{sub c}]{sub n} and [GT{sub c}]{sub n}-G type. Chain shortening plus enrichment in trans conformers favour the crystallization process which occurs earlier and faster with modification in crystal populations. Additional physicochemical steps are necessary to preserve the main benefits of PET. - Highlights: • Combination of multiple techniques to characterize the effects of recycling in PET. • Cleavage of ester bonds reduced viscosity, Mw, toughness in mechanical recycled PET. • Virgin, mechanical recycled and commercial recycled PET differ in crystal populations. • Cyclic oligomers [GT{sub c}]{sub n} and [GT{sub c}]{sub n}-G increase from the fourth extrusion cycle onwards.

Dual recycling for GEO 600

International Nuclear Information System (INIS)

Grote, H; Freise, A; Malec, M; Heinzel, G; Willke, B; Lueck, H; Strain, K A; Hough, J; Danzmann, K

2004-01-01

Dual recycling is the combination of signal recycling and power recycling; both optical techniques improve the shot-noise-limited sensitivity of interferometric gravitational-wave detectors. In addition, signal recycling can reduce the loss of light power due to imperfect interference and allows us, in principle, to beat the standard quantum limit. The interferometric gravitational-wave detector GEO 600 is the first of the kilometre-scale detectors to use signal recycling. We have recently equipped the detector with a signal-recycling mirror with a transmittance of 1%. In this paper, we present details of the detector commissioning and the first locks of the dual-recycled interferometer

Present status of recycling waste mobile phones in China: a review.

Science.gov (United States)

Li, Jingying; Ge, Zhongying; Liang, Changjin; An, Ni

2017-07-01

A large number of waste mobile phones have already been generated and are being generated. Various countries around the world have all been positively exploring the way of recycling and reuse when facing such a large amount of waste mobile phones. In some countries, processing waste mobile phones has been forming a complete industrial chain, which can not only recycle waste mobile phones to reduce their negative influence on the environment but also turn waste into treasure to acquire economic benefits dramatically. However, the situation of recycling waste mobile phones in China is not going well. Waste mobile phones are not formally covered by existing regulations and policies for the waste electric and electronic equipment in China. In order to explore an appropriate system to recover waste mobile phones, the mobile phone production and the amount of waste mobile phones are introduced in this paper, and status of waste mobile phones recycling is described; then, the disposal technology of electronic waste that would be most likely to be used for processing of electronic waste in industrial applications in the near future is reviewed. Finally, rationalization proposals are put forward based on the current recovery status of waste mobile phones for the purpose of promoting the development of recycling waste mobile phones in developing countries with a special emphasis on China.

Nramp1 promotes efficient macrophage recycling of iron following erythrophagocytosis in vivo

OpenAIRE

Soe-Lin, Shan; Apte, Sameer S.; Andriopoulos, Billy; Andrews, Marc C.; Schranzhofer, Matthias; Kahawita, Tanya; Garcia-Santos, Daniel; Ponka, Prem

2009-01-01

Natural resistance-associated macrophage protein 1 (Nramp1) is a divalent metal transporter expressed exclusively in phagocytic cells. We hypothesized that macrophage Nramp1 may participate in the recycling of iron acquired from phagocytosed senescent erythrocytes. To evaluate the role of Nramp1 in vivo, the iron parameters of WT and KO mice were analyzed after acute and chronic induction of hemolytic anemia. We found that untreated KO mice exhibited greater serum transferrin saturation and s...

Towards sustainable consumption: A socio-economic analysis of household waste recycling outcomes in Hong Kong.

Science.gov (United States)

Lo, Alex Y; Liu, Shuwen

2018-05-15

Many high-density cities struggle to find space for disposing municipal solid waste. Hong Kong is one of these cities, seeking to scale up waste recovery efforts as an alternative to disposal. However, territory-wide recovery initiatives do not account for socio-economic variations across place, leading to mixed outcomes among diverse communities. This study aims to investigate socio-economic effects on recycling behavior in a sample of subsidized rental housing estates. It constitutes an improvement from previous studies by using the entire estate as a unit of analysis and analyzing actual recycling outcomes, which have received limited attention from researchers. The analysis focused on the volume of recyclables collected from 158 public housing estates in Hong Kong, with an average population of 12,285. Results suggest that recycling outcomes vary with a limited set of socio-economic factors. Housing estates managed by a private property management company and populated by better off households collected more recyclables from their residents. Measures of absolute and relative recycling intensity achieved similar results. The findings will be useful for identifying residential communities requiring additional support for promoting waste separation and recycling. Differentiated policies for economically disadvantaged communities are warranted. Copyright © 2018 Elsevier Ltd. All rights reserved.

Recycle Alaska: Reduce, Reuse, Recycle. Activities Handbook, Teacher's Guide, and Student Worksheets.

Science.gov (United States)

Alaska State Dept. of Education, Juneau.

Recycling is a very important aspect of conserving the environment for future generations. This guide addresses the topic of litter prevention for the Alaskan environment and contains 42 activities. Activity topics covered include Natural Cycles, Human Interruption of Natural Cycles, Reduce, Reuse, Recycle and Recycled Classroom. Grade level,…

The Diffusion Effect of MSW Recycling

Directory of Open Access Journals (Sweden)

Yi-Tui Chen

2017-12-01

Full Text Available The purpose of this paper is to compare the recycling performance for some waste fractions selected including food waste, bulk waste, paper, metal products, plastics/rubber and glass products and then to develop some directions for the future improvements. The priority of each waste fraction for recycling is also analyzed by using an importance-performance analysis. Traditionally, the recycling rate that is calculated by the ratio of waste recycled to waste collected is used as an indicator to measure recycling performance. Due to a large variation among waste fractions in municipal solid waste (MSW, the recycling rate cannot reflect the actual recycling performance. The ceiling of recycling rate for each waste fraction estimated from the diffusion models is incorporated into a model to calculate recycling performance. The results show that (1 the diffusion effect exists significantly for the recycling of most recyclables but no evidence is found to support the diffusion effect for the recycling of food waste and bulk waste; (2 the recycling performance of waste metal products ranks the top, compared to waste paper, waste glass and other waste fractions; (3 furthermore, an importance-performance analysis (IPA is employed to analyze the priority of recycling programs and thus this paper suggests that the recycling of food waste should be seen as the most priority item to recycle.

CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL

NARCIS (Netherlands)

Bartuzi, Paulina; Billadeau, Daniel D.; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H.; Huijkman, Nicolette; Kloosterhuis, Niels; Van der Molen, Henk; Brufau, Gemma; Groen, Albert K.; Elliott, Alison M.; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D.; Burstein, Ezra; Hofker, Marten H.; van de Sluis, Bart

2016-01-01

The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal

CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL

NARCIS (Netherlands)

Bartuzi, Paulina; Billadeau, Daniel D.; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H.; Huijkman, Nicolette; Kloosterhuis, Niels; van der Molen, Henk; Brufau, Gemma; Groen, Albert K.; Elliott, Alison M.; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D.; Burstein, Ezra; Hofker, Marten H.; van de Sluis, Bart

The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal

Kinesin-3 and dynein cooperate in long-range retrograde endosome motility along a nonuniform microtubule array

NARCIS (Netherlands)

Schuster, M.; Kilaru, S.; Fink, G.; Collemare, J.A.R.; Roger, Y.; Steinberg, G.

2011-01-01

The polarity of microtubules (MTs) determines the motors for intracellular motility, with kinesins moving to plus ends and dynein to minus ends. In elongated cells of Ustilago maydis, dynein is thought to move early endosomes (EEs) toward the septum (retrograde), whereas kinesin-3 transports them to

Evaluating iodide recycling inhibition as a novel molecular initiating event for thyroid axis disruption

Science.gov (United States)

The enzyme iodotyrosine deiodinase (dehalogenase, IYD) catalyzes iodide recycling and promotes iodide retention in thyroid follicular cells. Loss of function or chemical inhibition of IYD reduces available iodide for thyroid hormone synthesis, which leads to hormone insufficiency...

α1B-Adrenergic Receptors Differentially Associate with Rab Proteins during Homologous and Heterologous Desensitization

Science.gov (United States)

Castillo-Badillo, Jean A.; Sánchez-Reyes, Omar B.; Alfonzo-Méndez, Marco A.; Romero-Ávila, M. Teresa; Reyes-Cruz, Guadalupe; García-Sáinz, J. Adolfo

2015-01-01

Internalization of G protein-coupled receptors can be triggered by agonists or by other stimuli. The process begins within seconds of cell activation and contributes to receptor desensitization. The Rab GTPase family controls endocytosis, vesicular trafficking, and endosomal fusion. Among their remarkable properties is the differential distribution of its members on the surface of various organelles. In the endocytic pathway, Rab 5 controls traffic from the plasma membrane to early endosomes, whereas Rab 4 and Rab 11 regulate rapid and slow recycling from early endosomes to the plasma membrane, respectively. Moreover, Rab 7 and Rab 9 regulate the traffic from late endosomes to lysosomes and recycling to the trans-Golgi. We explore the possibility that α1B-adrenergic receptor internalization induced by agonists (homologous) and by unrelated stimuli (heterologous) could involve different Rab proteins. This possibility was explored by Fluorescence Resonance Energy Transfer (FRET) using cells coexpressing α1B-adrenergic receptors tagged with the red fluorescent protein, DsRed, and different Rab proteins tagged with the green fluorescent protein. It was observed that when α1B-adrenergic receptors were stimulated with noradrenaline, the receptors interacted with proteins present in early endosomes, such as the early endosomes antigen 1, Rab 5, Rab 4, and Rab 11 but not with late endosome markers, such as Rab 9 and Rab 7. In contrast, sphingosine 1-phosphate stimulation induced rapid and transient α1B-adrenergic receptor interaction of relatively small magnitude with Rab 5 and a more pronounced and sustained one with Rab 9; interaction was also observed with Rab 7. Moreover, the GTPase activity of the Rab proteins appears to be required because no FRET was observed when dominant-negative Rab mutants were employed. These data indicate that α1B-adrenergic receptors are directed to different endocytic vesicles depending on the desensitization type (homologous vs

Recycle and reuse of radioactive scrap metals within the department of energy

International Nuclear Information System (INIS)

Adams, V.; Murphie, W.; Gresalfi, M.

2000-01-01

The United States Department of Energy (DOE) National Center of Excellence for Metals Recycle (NMR) is pursuing recycle and reuse alternatives to burial of radioactive scrap metal. This approach is being implemented in a safe and environmentally sound manner, while significantly lowering dis-positioning cost and accelerating cleanup activities. This paper will define the NMR's success to date in promoting safe and cost effective recycle and reuse strategies for DOE's excess metals, through the use of case studies. The paper will also present actual volumes of metal moved by DOE into restricted and unrestricted uses since 1997. In addition, this paper will discuss the principle underlying the Three Building Decommissioning and Decontamination (D and D) Project in Oak Ridge, Tennessee. In January 2000, the Secretary of Energy placed a moratorium on the unrestricted release of volumetrically contaminated metals from the DOE sites. Pursuant to that moratorium, the Secretary also established a ''Re-Use and Recycling Task Force'' to conduct a review of DOE policies regarding the management and release of all materials for recycle and reuse from DOE facilities. This task force was charged to develop a set of recommendations to ensure the protection of public health and the environment, openness and public trust, and fiscal responsibility. This paper will present an overview of the DOE's present range of recycle and reuse alternatives to disposal, as practiced by the NMR, and discuss the policy and issues associated with the task force mission. (authors)

Compression Molding of Composite of Recycled HDPE and Recycled Tire Particles

Science.gov (United States)

Liu, Ping; Waskom, Tommy L.; Chen, Zhengyu; Li, Yanze; Peng, Linda

1996-01-01

Plastic and rubber recycling is an effective means of reducing solid waste to the environment and preserving natural resources. A project aimed at developing a new composite material from recycled high density polyethylene (HDPE) and recycled rubber is currently being conducted at Eastern Illinois University. The recycled plastic pellets with recycled rubber particles are extruded into some HDPE/rubber composite strands. The strand can be further cut into pellets that can be used to fabricate other material forms or products. This experiment was inspired by the above-mentioned research activity. In order to measure Durometer hardness of the extruded composite, a specimen with relatively large dimensions was needed. Thus, compression molding was used to form a cylindrical specimen of 1 in. diameter and 1 in. thickness. The initial poor quality of the molded specimen prompted a need to optimize the processing parameters such as temperature, holding time, and pressure. Design of experiment (DOE) was used to obtain optimum combination of the parameters.

Certified Electronics Recyclers

Science.gov (United States)

Learn how EPA encourages all electronics recyclers become certified by demonstrating to an accredited, independent third-party auditor and that they meet specific standards to safely recycle and manage electronics.

Electron beam radiation effects on recycled polyamide-6

International Nuclear Information System (INIS)

Evora, Maria Cecilia; Silva, Leonardo G. de Andrade e

2001-01-01

Applications of electron beam processing in the treatment of polymers are commonly used. The interaction of high energy radiation with polymers may cause permanent modifications in the polymer's physicochemical structure. The induced modifications may result in degradation of the polymer or in improvement of its properties (crosslinking), which are simultaneous and competing processes, depending on the radiation dose utilized. Crosslinking occurs more readily in the polymer's amorphous content and this process makes the glass transition temperature (Tg) of the polymers to increase. Successive recycling cycles promote changes in polymers properties, such as breaking of structure, molecular weight reduction, melt index increase and mechanical resistance reduction. The polyamide-6 resin was recycled for three successive recycling cycles and thi polyamide-6 specimens were molded by the process of injection molding. These specimens were irradiated at the Nuclear Energetic Research Institute (IPEN) radiation facility, on a JOB 188 model accelerator, with a 1.5 MeV electron beam, doses of 200, 300, 400, 500 and 600 kGy, and dose rate of 22.61 kGy/s. The DMA tests were performed using DMA-983 equipment from TA Instruments and two heatings were adopted in order to eliminate the moisture absorption. The X-ray diffraction analysis wa carried out at the Philips PW 1830 model equipment

Genetic reconstitution of the human Adenovirus type 2 temperature-sensitive 1 mutant defective in endosomal escape

Directory of Open Access Journals (Sweden)

Gastaldelli Michele

2009-10-01

Full Text Available Abstract Human Adenoviruses infect the upper and lower respiratory tracts, the urinary and digestive tracts, lymphoid systems and heart, and give rise to epidemic conjunctivitis. More than 51 human serotypes have been identified to-date, and classified into 6 species A-F. The species C Adenoviruses Ad2 and Ad5 (Ad2/5 cause upper and lower respiratory disease, but how viral structure relates to the selection of particular infectious uptake pathways is not known. An adenovirus mutant, Ad2-ts1 had been isolated upon chemical mutagenesis in the past, and shown to have unprocessed capsid proteins. Ad2-ts1 fails to package the viral protease L3/p23, and Ad2-ts1 virions do not efficiently escape from endosomes. It had been suggested that the C22187T point mutation leading to the substitution of the conserved proline 137 to leucine (P137L in the L3/p23 protease was at least in part responsible for this phenotype. To clarify if the C22187T mutation is necessary and sufficient for the Ad2-ts1 phenotype, we sequenced the genes encoding the structural proteins of Ad2-ts1, and confirmed that the Ad2-ts1 DNA carries the point mutation C22187T. Introduction of C22187T to the wild-type Ad2 genome in a bacterial artificial chromosome (Ad2-BAC gave Ad2-BAC46 virions with the full Ad2-ts1 phenotype. Reversion of Ad2-BAC46 gave wild-type Ad2 particles indicating that P137L is necessary and sufficient for the Ad2-ts1 phenotype. The kinetics of Ad2-ts1 uptake into cells were comparable to Ad2 suggesting similar endocytic uptake mechanisms. Surprisingly, infectious Ad2 or Ad5 but not Ad2-ts1 uptake required CALM (clathrin assembly lymphoid myeloid protein, which controls clathrin-mediated endocytosis and membrane transport between endosomes and the trans-Golgi-network. The data show that no other mutations than P137L in the viral protease are necessary to give rise to particles that are defective in capsid processing and endosomal escape. This provides a basis for

Accumulation of dsRNA in endosomes contributes to inefficient RNA interference in the fall armyworm, Spodoptera frugiperda.

Science.gov (United States)

Yoon, June-Sun; Gurusamy, Dhandapani; Palli, Subba Reddy

2017-11-01

RNA interference (RNAi) efficiency varies among insects studied. The barriers for successful RNAi include the presence of double-stranded ribonucleases (dsRNase) in the lumen and hemolymph that could potentially digest double-stranded RNA (dsRNA) and the variability in the transport of dsRNA into and within the cells. We recently showed that the dsRNAs are transported into lepidopteran cells, but they are not processed into small interference RNAs (siRNAs) because they are trapped in acidic bodies. In the current study, we focused on the identification of acidic bodies in which dsRNAs accumulate in Sf9 cells. Time-lapse imaging studies showed that dsRNAs enter Sf9 cells and accumulate in acidic bodies within 20 min after their addition to the medium. CypHer-5E-labeled dsRNA also accumulated in the midgut and fat body dissected from Spodoptera frugiperda larvae with similar patterns observed in Sf9 cells. Pharmacological inhibitor assays showed that the dsRNAs use clathrin mediated endocytosis pathway for transport into the cells. We investigated the potential dsRNA accumulation sites employing LysoTracker and double labeling experiments using the constructs to express a fusion of green fluorescence protein with early or late endosomal marker proteins and CypHer-5E-labeled dsRNA. Interestingly, CypHer-5E-labeled dsRNA accumulated predominantly in early and late endosomes. These data suggest that entrapment of internalized dsRNA in endosomes is one of the major factors contributing to inefficient RNAi response in lepidopteran insects. Copyright © 2017 Elsevier Ltd. All rights reserved.

PET and Recycling

Directory of Open Access Journals (Sweden)

Funda Sevencan

2007-08-01

Full Text Available This review aims to clarify the need of decreasing the environmental effects caused by human and draw attention to the increasing environmental effects of plastics wastes. Plastics consist of organic molecules with high density molecules or polymers. Main resources of plastics are the residue of oil rafineries. Several advantages of plastics, have increased the usage continuously. Polyethylene Terephthalate (PET is the most commonly used plastics. PET is used to protect food, drinking water, fruit juice, alcoholic beverage, and food packing films. By the increasing interest on the environmental effects of plastic wastes, concerns on the recyclable packing materials also grew up. Also the daily use of recyclable containers consisting PET have increased. There are five steps for recycling of plastics. These steps are; using large amounts of plastics, collecting them in a big center, classifying and sorting the plastics, reproducing the polymers and obtaining new products with melted plastics. Providing a healthy recycling of plastics, the consumers should have knowledge and responsibility. The consumer should know what he/she has to do before putting the plastics in the recycling containers. Recycling containers and bags should be placed near the sources of plastic wastes. Consequently, the plastic wastes and environmental problems they cause will be on the agenda in future. [TAF Prev Med Bull. 2007; 6(4: 307-312

PET and Recycling

Directory of Open Access Journals (Sweden)

Funda Sevencan

2007-08-01

Full Text Available This review aims to clarify the need of decreasing the environmental effects caused by human and draw attention to the increasing environmental effects of plastics wastes. Plastics consist of organic molecules with high density molecules or polymers. Main resources of plastics are the residue of oil rafineries. Several advantages of plastics, have increased the usage continuously. Polyethylene Terephthalate (PET is the most commonly used plastics. PET is used to protect food, drinking water, fruit juice, alcoholic beverage, and food packing films. By the increasing interest on the environmental effects of plastic wastes, concerns on the recyclable packing materials also grew up. Also the daily use of recyclable containers consisting PET have increased. There are five steps for recycling of plastics. These steps are; using large amounts of plastics, collecting them in a big center, classifying and sorting the plastics, reproducing the polymers and obtaining new products with melted plastics. Providing a healthy recycling of plastics, the consumers should have knowledge and responsibility. The consumer should know what he/she has to do before putting the plastics in the recycling containers. Recycling containers and bags should be placed near the sources of plastic wastes. Consequently, the plastic wastes and environmental problems they cause will be on the agenda in future. [TAF Prev Med Bull 2007; 6(4.000: 307-312

Life Cycle Analysis of High Quality Recycled Aggregate Produced byHeating and Rubbing Method

Science.gov (United States)

Shima, Hirokazu; Matsuhashi, Ryuji; Yoshida, Yoshikuni; Tateyashiki, Hisashi

Most of demolished concrete is recycled as road subbase, but its generation is expected to increase rapidly and exceed the demand of road subbase in a near future. To promote the recycling of concrete, the technology to produce high quality recycled aggregate by the heating and rubbing method has been developed. In this method, demolished concrete is heated up to about 300°C in a heater to make cement paste brittle with its dehydration. The heated concrete is then rubbed in two mills to recover the recycled aggregate, while the paste is removed from the surface of aggregate and collected as cement fine powder. In this method, much energy is consumed to heat and rub concrete; however, the cement fine powder is utilized for a soil stabilizer and cement raw materials, so that the environmental load is reduced in cement manufacturing. The life cycle analysis of the recycled aggregate is carried out to evaluate this technology. As a result, the life cycle CO2 is a negative value because the deduction of CO2 emission in cement manufacturing by the powder is much large. This technology is proved to be very effective to reduce CO2.

Reusing and recycling in Saskatchewan: Environmental benefits of reusing and recycling

Energy Technology Data Exchange (ETDEWEB)

1994-01-01

After an introduction explaining the environmental benefits of reusing and recycling, as well as providing suggestions on minimizing waste and conserving energy, a directory of recyclers and handlers of various kinds of waste in Saskatchewan is presented. Names, addresses/telephone numbers, and types of materials accepted are given for recyclers of animal products, clothing or textiles, glass, compostable materials, industrial hardware, metals, office products, paper, plastic, and tires. Collection depots in the SARCAN recycling program for beverage containers are listed, giving town name, address, hours of operation, and telephone number. Receivers of waste dangerous goods are listed under the categories of ozone-depleting substances, waste batteries, solvents, lubricating oils and oil filters, paint, flammable liquids, antifreeze, drycleaning waste, and miscellaneous.

Recycling of Paper and Cardboard

DEFF Research Database (Denmark)

Christensen, Thomas Højlund; Damgaard, Anders

2011-01-01

waste. Recycling of paper and cardboard production waste and postconsumer waste has a long history in the pulp and paper industry. The recycled material now makes up more than half of the raw material used in European pulp and paper industry (ERPC, 2004). This chapter describes briefly how paper...... and cardboard are produced and how waste paper is recycled in the industry. Quality requirements and use of recycled products are discussed, as are the resource and environmental issues of paper recycling....

Water Recycling in Australia

Directory of Open Access Journals (Sweden)

Ross Young

2011-09-01

Full Text Available Australia is the driest inhabited continent on earth and, more importantly, experiences the most variable rainfall of all the continents on our planet. The vast majority of Australians live in large cities on the coast. Because wastewater treatments plants were all located near the coast, it was thought that large scale recycling would be problematic given the cost of infrastructure and pumping required to establish recycled water schemes. This all changed when Australia experienced a decade of record low rainfall and water utilities were given aggressive targets to increase the volume of water recycled. This resulted in recycled water being accepted as a legitimate source of water for non-drinking purposes in a diversified portfolio of water sources to mitigate climate risk. To ensure community support for recycled water, Australia lead the world in developing national guidelines for the various uses of recycled water to ensure the protection of public health and the environment. Australia now provides a great case study of the developments in maximizing water recycling opportunities from policy, regulatory and technological perspectives. This paper explores the evolution in thinking and how approaches to wastewater reuse has changed over the past 40 years from an effluent disposal issue to one of recognizing wastewater as a legitimate and valuable resource. Despite recycled water being a popular choice and being broadly embraced, the concept of indirect potable reuse schemes have lacked community and political support across Australia to date.

Recycling of concrete

International Nuclear Information System (INIS)

Halaszovich, S.

1988-01-01

The paper reviews potentials and problems of disposal or recycling of concrete removed from nuclear installations. Due to the difficulties in determining radioactivity limits that are compatible with utilization of recycled material in practice, a method is proposed that takes into account inhalation of dusts, as occurring during the reprocessing or recycling of the concrete, for instance in road building. This method is based on the maximum permissible radioactivity uptake by inhalation of a nuclide mixture of unknown composition. (RB) [de

Myo1c regulates lipid raft recycling to control cell spreading, migration and Salmonella invasion.

Science.gov (United States)

Brandstaetter, Hemma; Kendrick-Jones, John; Buss, Folma

2012-04-15

A balance between endocytosis and membrane recycling regulates the composition and dynamics of the plasma membrane. Internalization and recycling of cholesterol- and sphingolipid-enriched lipid rafts is an actin-dependent process that is mediated by a specialized Arf6-dependent recycling pathway. Here, we identify myosin1c (Myo1c) as the first motor protein that drives the formation of recycling tubules emanating from the perinuclear recycling compartment. We demonstrate that the single-headed Myo1c is a lipid-raft-associated motor protein that is specifically involved in recycling of lipid-raft-associated glycosylphosphatidylinositol (GPI)-linked cargo proteins and their delivery to the cell surface. Whereas Myo1c overexpression increases the levels of these raft proteins at the cell surface, in cells depleted of Myo1c function through RNA interference or overexpression of a dominant-negative mutant, these tubular transport carriers of the recycling pathway are lost and GPI-linked raft markers are trapped in the perinuclear recycling compartment. Intriguingly, Myo1c only selectively promotes delivery of lipid raft membranes back to the cell surface and is not required for recycling of cargo, such as the transferrin receptor, which is mediated by parallel pathways. The profound defect in lipid raft trafficking in Myo1c-knockdown cells has a dramatic impact on cell spreading, cell migration and cholesterol-dependent Salmonella invasion; processes that require lipid raft transport to the cell surface to deliver signaling components and the extra membrane essential for cell surface expansion and remodeling. Thus, Myo1c plays a crucial role in the recycling of lipid raft membrane and proteins that regulate plasma membrane plasticity, cell motility and pathogen entry.

Drosophila VAMP7 regulates Wingless intracellular trafficking.

Science.gov (United States)

Gao, Han; He, Fang; Lin, Xinhua; Wu, Yihui

2017-01-01

Drosophila Wingless (Wg) is a morphogen that determines cell fate during development. Previous studies have shown that endocytic pathways regulate Wg trafficking and signaling. Here, we showed that loss of vamp7, a gene required for vesicle fusion, dramatically increased Wg levels and decreased Wg signaling. Interestingly, we found that levels of Dally-like (Dlp), a glypican that can interact with Wg to suppress Wg signaling at the dorsoventral boundary of the Drosophila wing, were also increased in vamp7 mutant cells. Moreover, Wg puncta in Rab4-dependent recycling endosomes were Dlp positive. We hypothesize that VAMP7 is required for Wg intracellular trafficking and the accumulation of Wg in Rab4-dependent recycling endosomes might affect Wg signaling.

Aluminium beverage can recycling

Energy Technology Data Exchange (ETDEWEB)

Lewinski, A von

1985-08-01

Canned beverages have become a controversial issue in this era of ecological sensitivity. METALL has already discussed the problem of can recycling. The present article discusses the technical aspects of aluminium can recycling. Two further articles will follow on aluminium can recycling in North America and on the results of European pilot projects.

Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast.

Directory of Open Access Journals (Sweden)

Ayako Kita

Full Text Available BACKGROUND: We had previously identified the mutant allele of apm1(+ that encodes a homolog of the mammalian µ1A subunit of the clathrin-associated adaptor protein-1 (AP-1 complex, and we demonstrated the role of Apm1 in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we isolated rho3(+, which encodes a Rho-family small GTPase, an important regulator of exocystosis, as a multicopy-suppressor of the temperature-sensitive growth of the apm1-1 mutant cells. Overexpression of Rho3 suppressed the Cl(- sensitivity and immunosuppressant sensitivity of the apm1-1 mutant cells. Overexpression of Rho3 also suppressed the fragmentation of vacuoles, and the accumulation of v-SNARE Syb1 in Golgi/endosomes and partially suppressed the defective secretion associated with apm1-deletion cells. Notably, electron microscopic observation of the rho3-deletion cells revealed the accumulation of abnormal Golgi-like structures, vacuole fragmentation, and accumulation of secretory vesicles; these phenotypes were very similar to those of the apm1-deletion cells. Furthermore, the rho3-deletion cells and apm1-deletion cells showed very similar phenotypic characteristics, including the sensitivity to the immunosuppressant FK506, the cell wall-damaging agent micafungin, Cl(-, and valproic acid. Green fluorescent protein (GFP-Rho3 was localized at Golgi/endosomes as well as the plasma membrane and division site. Finally, Rho3 was shown to form a complex with Apm1 as well as with other subunits of the clathrin-associated AP-1 complex in a GTP- and effector domain-dependent manner. CONCLUSIONS/SIGNIFICANCE: Taken together, our findings reveal a novel role of Rho3 in the regulation of Golgi/endosome trafficking and suggest that clathrin-associated adaptor protein-1 and Rho3 co-ordinate in intracellular transport in fission yeast. To the best of our knowledge, this study provides the first evidence

The Fernald Waste Recycling Program

International Nuclear Information System (INIS)

Motl, G.P.

1993-01-01

Recycling is considered a critical component of the waste disposition strategy at the Fernald Plant. It is estimated that 33 million cubic feet of waste will be generated during the Fernald cleanup. Recycling some portion of this waste will not only conserve natural resources and disposal volume but will, even more significantly, support the preservation of existing disposition options such as off-site disposal or on-site storage. Recognizing the strategic implications of recycling, this paper outlines the criteria used at Fernald to make recycle decisions and highlights several of Fernald's current recycling initiatives

Localization of the AP-3 adaptor complex defines a novel endosomal exit site for lysosomal membrane proteins

NARCIS (Netherlands)

Peden, A.A.; Oorschot, V.; Hesser, B.A.; Austin, C.D.; Scheller, R.H.; Klumperman, J.

2004-01-01

The adaptor protein (AP) 3 adaptor complex has been implicated in the transport of lysosomal membrane proteins, but its precise site of action has remained controversial. Here, we show by immuno-electron microscopy that AP-3 is associated with budding profiles evolving from a tubular endosomal

The recycling is moving

CERN Multimedia

GS Department

2011-01-01

The recycling site currently situated near building 133 has been transferred to the car park of building 156. The site is identified by the sign “RECYCLING” and the above logo. In this new, more accessible site, you will find recycling bins for the following waste: PET (recyclable plastic bottles); Aluminium cans; Nespresso coffee capsules.  

A tiered approach to evaluate an iodine recycling inhibition adverse outcome pathway (AOP) in amphibians

Science.gov (United States)

The enzyme iodotyrosine deiodinase (dehalogenase, IYD) catalyzes iodide recycling and promotes iodide retention in thyroid follicular cells. Loss of function or chemical inhibition of IYD reduces thyroid hormone synthesis, which leads to insufficiency in tissues and subsequent ne...

Transport, metabolism, and endosomal trafficking-dependent regulation of intestinal fructose absorption

Science.gov (United States)

Patel, Chirag; Douard, Veronique; Yu, Shiyan; Gao, Nan; Ferraris, Ronaldo P.

2015-01-01

Dietary fructose that is linked to metabolic abnormalities can up-regulate its own absorption, but the underlying regulatory mechanisms are not known. We hypothesized that glucose transporter (GLUT) protein, member 5 (GLUT5) is the primary fructose transporter and that fructose absorption via GLUT5, metabolism via ketohexokinase (KHK), as well as GLUT5 trafficking to the apical membrane via the Ras-related protein-in-brain 11 (Rab11)a-dependent endosomes are each required for regulation. Introducing fructose but not lysine and glucose solutions into the lumen increased by 2- to 10-fold the heterogeneous nuclear RNA, mRNA, protein, and activity levels of GLUT5 in adult wild-type mice consuming chow. Levels of GLUT5 were >100-fold that of candidate apical fructose transporters GLUTs 7, 8, and 12 whose expression, and that of GLUT 2 and the sodium-dependent glucose transporter protein 1 (SGLT1), was not regulated by luminal fructose. GLUT5-knockout (KO) mice exhibited no facilitative fructose transport and no compensatory increases in activity and expression of SGLT1 and other GLUTs. Fructose could not up-regulate GLUT5 in GLUT5-KO, KHK-KO, and intestinal epithelial cell-specific Rab11a-KO mice. The fructose-specific metabolite glyceraldehyde did not increase GLUT5 expression. GLUT5 is the primary transporter responsible for facilitative absorption of fructose, and its regulation specifically requires fructose uptake and metabolism and normal GLUT5 trafficking to the apical membrane.—Patel, C., Douard, V., Yu, S., Gao, N., Ferraris, R. P. Transport, metabolism, and endosomal trafficking-dependent regulation of intestinal fructose absorption. PMID:26071406

Rabies virus co-localizes with early (Rab5) and late (Rab7) endosomal proteins in neuronal and SH-SY5Y cells.

Science.gov (United States)

Ahmad, Waqas; Li, Yingying; Guo, Yidi; Wang, Xinyu; Duan, Ming; Guan, Zhenhong; Liu, Zengshan; Zhang, Maolin

2017-06-01

Rabies virus (RABV) is a highly neurotropic virus that follows clathrin-mediated endocytosis and pH-dependent pathway for trafficking and invasion into endothelial cells. Early (Rab5, EEA1) and late (Rab7, LAMP1) endosomal proteins play critical roles in endosomal sorting, maturity and targeting various molecular cargoes, but their precise functions in the early stage of RABV neuronal infection remain elusive. In this study, the relationship between enigmatic entry of RABV with these endosomal proteins into neuronal and SH-SY5Y cells was investigated. Immunofluorescence, TCID 50 titers, electron microscopy and western blotting were carried out to determine the molecular interaction of the nucleoprotein (N) of RABV with early or late endosomal proteins in these cell lines. The expression of N was also determined by down-regulating Rab5 and Rab7 in both cell lines through RNA interference. The results were indicative that N proficiently colocalized with Rab5/EEA1 and Rab7/LAMP1 in both cell lines at 24 and 48 h post-infection, while N titers significantly decreased in early infection of RABV. Down-regulation of Rab5 and Rab7 did not inhibit N expression, but it prevented productive infection via blocking the normal trafficking of RABV in a low pH environment. Ultrathin sections of cells studied by electron microscope also verified the close association of RABV with Rab5 and Rab7 in neurons. From the data it was concluded that primary entry of RABV strongly correlates with the kinetics of Rab-proteins present on early and late vesicles, which provides helpful clues to explain the early events of RABV in nerve cells.

Tire Recycling

Science.gov (United States)

1997-01-01

Cryopolymers, Inc. tapped NASA expertise to improve a process for recycling vehicle tires by converting shredded rubber into products that can be used in asphalt road beds, new tires, hoses, and other products. In conjunction with the Southern Technology Applications Center and Stennis Space Center, NASA expertise in cryogenic fuel-handling needed for launch vehicle and spacecraft operations was called upon to improve the recycling concept. Stennis advised Cryopolymers on the type of equipment required, as well as steps to reduce the amount of liquid nitrogen used in the process. They also guided the company to use more efficient ways to control system hardware. It is estimated that more than 300 million tires nationwide are produced per year. Cryopolymers expects to reach a production rate of 5,000 tires recycled per day.

The small G protein Arl5 contributes to endosome-to-Golgi traffic by aiding the recruitment of the GARP complex to the Golgi

Directory of Open Access Journals (Sweden)

Cláudia Rosa-Ferreira

2015-03-01

Full Text Available The small G proteins of the Arf family play critical roles in membrane trafficking and cytoskeleton organization. However, the function of some members of the family remains poorly understood including Arl5 which is widely conserved in eukaryotes. Humans have two closely related Arl5 paralogues (Arl5a and Arl5b, and both Arl5a and Arl5b localize to the trans-Golgi with Arl5b being involved in retrograde traffic from endosomes to the Golgi apparatus. To investigate the function of Arl5, we have used Drosophila melanogaster as a model system. We find that the single Arl5 orthologue in Drosophila also localizes to the trans-Golgi, but flies lacking the Arl5 gene are viable and fertile. By using both liposome and column based affinity chromatography methods we find that Arl5 interacts with the Golgi-associated retrograde protein (GARP complex that acts in the tethering of vesicles moving from endosomes to the trans-Golgi network (TGN. In Drosophila tissues the GARP complex is partially displaced from the Golgi when Arl5 is absent, and the late endosomal compartment is enlarged. In addition, in HeLa cells GARP also becomes cytosolic upon depletion of Arl5b. These phenotypes are consistent with a role in endosome-to-Golgi traffic, but are less severe than loss of GARP itself. Thus it appears that Arl5 is one of the factors that directs the recruitment of the GARP complex to the trans-Golgi, and this function is conserved in both flies and humans.

Recycle Glass in Foam Glass Production

DEFF Research Database (Denmark)

Petersen, Rasmus Rosenlund; König, Jakob; Yue, Yuanzheng

The foam glass industry turn recycle glass into heat insulating building materials. The foaming process is relative insensitive to impurities in the recycle glass. It is therefore considered to play an important role in future glass recycling. We show and discuss trends of use of recycled glasses...... in foam glass industry and the supply sources and capacity of recycle glass....

Chemical-mineralogical characterisation of coarse recycled concrete aggregate

International Nuclear Information System (INIS)

Limbachiya, M.C.; Marrocchino, E.; Koulouris, A.

2007-01-01

The construction industry is now putting greater emphasis than ever before on increasing recycling and promoting more sustainable waste management practices. In keeping with this approach, many sectors of the industry have actively sought to encourage the use of recycled concrete aggregate (RCA) as an alternative to primary aggregates in concrete production. The results of a laboratory experimental programme aimed at establishing chemical and mineralogical characteristics of coarse RCA and its likely influence on concrete performance are reported in this paper. Commercially produced coarse RCA and natural aggregates (16-4 mm size fraction) were tested. Results of X-ray fluorescence (XRF) analyses showed that original source of RCA had a negligible effect on the major elements and a comparable chemical composition between recycled and natural aggregates. X-ray diffraction (XRD) analyses results indicated the presence of calcite, portlandite and minor peaks of muscovite/illite in recycled aggregates, although they were directly proportioned to their original composition. The influence of 30%, 50%, and 100% coarse RCA on the chemical composition of equal design strength concrete has been established, and its suitability for use in a concrete application has been assessed. In this work, coarse RCA was used as a direct replacement for natural gravel in concrete production. Test results indicated that up to 30% coarse RCA had no effect on the main three oxides (SiO 2 , Al 2 O 3 and CaO) of concrete, but thereafter there was a marginal decrease in SiO 2 and increase in Al 2 O 3 and CaO contents with increase in RCA content in the mix, reflecting the original constituent's composition

Chemical-mineralogical characterisation of coarse recycled concrete aggregate.

Science.gov (United States)

Limbachiya, M C; Marrocchino, E; Koulouris, A

2007-01-01

The construction industry is now putting greater emphasis than ever before on increasing recycling and promoting more sustainable waste management practices. In keeping with this approach, many sectors of the industry have actively sought to encourage the use of recycled concrete aggregate (RCA) as an alternative to primary aggregates in concrete production. The results of a laboratory experimental programme aimed at establishing chemical and mineralogical characteristics of coarse RCA and its likely influence on concrete performance are reported in this paper. Commercially produced coarse RCA and natural aggregates (16-4 mm size fraction) were tested. Results of X-ray fluorescence (XRF) analyses showed that original source of RCA had a negligible effect on the major elements and a comparable chemical composition between recycled and natural aggregates. X-ray diffraction (XRD) analyses results indicated the presence of calcite, portlandite and minor peaks of muscovite/illite in recycled aggregates, although they were directly proportioned to their original composition. The influence of 30%, 50%, and 100% coarse RCA on the chemical composition of equal design strength concrete has been established, and its suitability for use in a concrete application has been assessed. In this work, coarse RCA was used as a direct replacement for natural gravel in concrete production. Test results indicated that up to 30% coarse RCA had no effect on the main three oxides (SiO2, Al2O3 and CaO) of concrete, but thereafter there was a marginal decrease in SiO2 and increase in Al2O3 and CaO contents with increase in RCA content in the mix, reflecting the original constituent's composition.

Auditing an intensive care unit recycling program.

Science.gov (United States)

Kubicki, Mark A; McGain, Forbes; O'Shea, Catherine J; Bates, Samantha

2015-06-01

The provision of health care has significant direct environmental effects such as energy and water use and waste production, and indirect effects, including manufacturing and transport of drugs and equipment. Recycling of hospital waste is one strategy to reduce waste disposed of as landfill, preserve resources, reduce greenhouse gas emissions, and potentially remain fiscally responsible. We began an intensive care unit recycling program, because a significant proportion of ICU waste was known to be recyclable. To determine the weight and proportion of ICU waste recycled, the proportion of incorrect waste disposal (including infectious waste contamination), the opportunity for further recycling and the financial effects of the recycling program. We weighed all waste and recyclables from an 11-bed ICU in an Australian metropolitan hospital for 7 non-consecutive days. As part of routine care, ICU waste was separated into general, infectious and recycling streams. Recycling streams were paper and cardboard, three plastics streams (polypropylene, mixed plastics and polyvinylchloride [PVC]) and commingled waste (steel, aluminium and some plastics). ICU waste from the waste and recycling bins was sorted into those five recycling streams, general waste and infectious waste. After sorting, the waste was weighed and examined. Recycling was classified as achieved (actual), potential and total. Potential recycling was defined as being acceptable to hospital protocol and local recycling programs. Direct and indirect financial costs, excluding labour, were examined. During the 7-day period, the total ICU waste was 505 kg: general waste, 222 kg (44%); infectious waste, 138 kg (27%); potentially recyclable waste, 145 kg (28%). Of the potentially recyclable waste, 70 kg (49%) was actually recycled (14% of the total ICU waste). In the infectious waste bins, 82% was truly infectious. There was no infectious contamination of the recycling streams. The PVC waste was 37% contaminated

Recycling of Metals

DEFF Research Database (Denmark)

Damgaard, Anders; Christensen, Thomas Højlund

2011-01-01

Metals like iron and aluminium are produced from mineral ore and used for a range of products, some of which have very short lifetimes and thus constitute a major fraction of municipal waste. Packaging in terms of cans, foils and containers are products with a short lifetime. Other products like...... appliances, vehicles and buildings, containing iron and aluminium metals, have long lifetimes before they end up in the waste stream. The recycling of production waste and postconsumer metals has a long history in the metal industry. Some metal smelters are today entirely based on scarp metals. This chapter...... describes briefly how iron and aluminium are produced and how scrap metal is recycled in the industry. Quality requirements and use of recycled products are discussed, as are the resource and environmental issues of metal recycling. Copper and other metals are also found in waste but in much smaller...

Nuclear reactor recyclation device

International Nuclear Information System (INIS)

Takigawa, Yukio; Chuma, Kazuto

1987-01-01

Purpose: To prevent the unevenness for the coolant flow rate even when abnormality occurs to one of recycling pumps. Constitution: A plurality of jet pumps disposed at an interval around the reactor core are divided circumferentially into two sets, and a pipeway is disposed to the outside of each pair including recycling pumps corresponding to each of the sets. The pipeway is connected to the recycling inlet of the jet pump by way of a manifold. The discharge portion of the recycling pumps of the loop pipeway are connected with each other by way of communication pipes, and a normally closed valve is disposed to the communication pipe and the normally closed valve of the communication pipe is opened upon detecting abnormality for one of the recycling pumps. Thus, if either one of the pair of recycling pumps shows abnormal state, coolants flows from the other of pipeway to the outside of the loop pipeway and coolants are supplied from all the jet pumps to the reactor core portion and, accordingly, the not-uniform flow rate can be prevented to eliminate undesired effect on the reactor core. (Kamimura, M.)

Recycling - Danish Waste Management Strategy

DEFF Research Database (Denmark)

Romann, Anne Funch; Thøgersen, John; Husmer, Lis

The report challanges recycling as the only waste handling strategy. The tonnes of recycled materials should not be the only goal - it is essential to minimize the waste production and focus on eliminating hazardous materials.......The report challanges recycling as the only waste handling strategy. The tonnes of recycled materials should not be the only goal - it is essential to minimize the waste production and focus on eliminating hazardous materials....

Length sensing and control of a Michelson interferometer with power recycling and twin signal recycling cavities.

Science.gov (United States)

Gräf, Christian; Thüring, André; Vahlbruch, Henning; Danzmann, Karsten; Schnabel, Roman

2013-03-11

The techniques of power recycling and signal recycling have proven as key concepts to increase the sensitivity of large-scale gravitational wave detectors by independent resonant enhancement of light power and signal sidebands within the interferometer. Developing the latter concept further, twin signal recycling was proposed as an alternative to conventional detuned signal recycling. Twin signal recycling features the narrow-band sensitivity gain of conventional detuned signal recycling but furthermore facilitates the injection of squeezed states of light, increases the detector sensitivity over a wide frequency band and requires a less complex detection scheme for optimal signal readout. These benefits come at the expense of an additional recycling mirror, thus increasing the number of degrees of freedom in the interferometer which need to be controlled.In this article we describe the development of a length sensing and control scheme and its successful application to a tabletop-scale power recycled Michelson interferometer with twin signal recycling. We were able to lock the interferometer in all relevant longitudinal degrees of freedom and thus laid the foundation for further investigations of this interferometer configuration to evaluate its viability for the application in gravitational wave detectors.

Involvement of the endosomal-lysosomal system correlates with regional pathology in Creutzfeldt-Jakob disease

DEFF Research Database (Denmark)

Kovács, Gábor G; Gelpi, Ellen; Ströbel, Thomas

2007-01-01

The endosomal-lysosomal system (ELS) has been suggested to play a role in the pathogenesis of prion diseases. The purpose of this study was to examine how experimental observations can be translated to human neuropathology and whether alterations of the ELS relate to neuropathologic changes...... correlate with regional pathology. Overloading of this system might impair the function of lysosomal enzymes and thus may mimic some features of lysosomal storage disorders. Udgivelsesdato: 2007-Jul...

Nuclear recycling: costs, savings, and safeguards

International Nuclear Information System (INIS)

Spinrad, B.I.

1985-01-01

This chapter discusses the economics, physical and chemical processes, and safety of nuclear fuel recycling. The spent fuel must be chemically reprocessed in order to recover uranium and plutonium. Topics considered include indifference costs, recycling in light water reactors (LWRs), plutonium in fast reactors, the choice between recycling and storage, safeguards, and weapons proliferation. It is shown that the economics of recycling nuclear fuel involves the actual costs and savings of the recycling operation in terms of money spent, made, and saved, and the impact of the recycling on the future cost of uranium

Obtaining multifunctional composites using styrofoam recycling: methylene blue adsorption

International Nuclear Information System (INIS)

Silva, T.H. da; Ferreira, B.F.; Faria, E.H. de; Ciuffi, K.J.; Calefi, P.S.

2011-01-01

There is high risk of environmental contamination by the textile industry if the generated waste is disposed into water streams and rivers. Another sector responsible for the generation of large amounts of waste is the styrofoam industry, since styrofoam is employed in various areas for packaging production. Therefore, today there is constant search for sustainable economic growth by means of materials recycling and reduction of residual pollutants. In this context, the present work aims to promote styrofoam recycling with its further utilization in the production of a kaolin-styrofoam composite for application as adsorbent of methylene blue originated from textile industry effluents. This adsorption is investigated in a column system, by exploiting the adsorptive properties of kaolin, such as large surface area and chemical and structural characteristics. The obtained materials were characterized by thermogravimetric analysis, X-ray diffraction, and UV-visible absorption spectroscopy. (author)

Benchmarking survey for recycling.

Energy Technology Data Exchange (ETDEWEB)

Marley, Margie Charlotte; Mizner, Jack Harry

2005-06-01

This report describes the methodology, analysis and conclusions of a comparison survey of recycling programs at ten Department of Energy sites including Sandia National Laboratories/New Mexico (SNL/NM). The goal of the survey was to compare SNL/NM's recycling performance with that of other federal facilities, and to identify activities and programs that could be implemented at SNL/NM to improve recycling performance.

GTPase ROP2 binds and promotes activation of target of rapamycin, TOR, in response to auxin.

Science.gov (United States)

Schepetilnikov, Mikhail; Makarian, Joelle; Srour, Ola; Geldreich, Angèle; Yang, Zhenbiao; Chicher, Johana; Hammann, Philippe; Ryabova, Lyubov A

2017-04-03

Target of rapamycin (TOR) promotes reinitiation at upstream ORFs (uORFs) in genes that play important roles in stem cell regulation and organogenesis in plants. Here, we report that the small GTPase ROP2, if activated by the phytohormone auxin, promotes activation of TOR, and thus translation reinitiation of uORF-containing mRNAs. Plants with high levels of active ROP2, including those expressing constitutively active ROP2 (CA-ROP2), contain high levels of active TOR ROP2 physically interacts with and, when GTP-bound, activates TOR in vitro TOR activation in response to auxin is abolished in ROP-deficient rop2 rop6 ROP4 RNAi plants. GFP-TOR can associate with endosome-like structures in ROP2-overexpressing plants, indicating that endosomes mediate ROP2 effects on TOR activation. CA-ROP2 is efficient in loading uORF-containing mRNAs onto polysomes and stimulates translation in protoplasts, and both processes are sensitive to TOR inhibitor AZD-8055. TOR inactivation abolishes ROP2 regulation of translation reinitiation, but not its effects on cytoskeleton or intracellular trafficking. These findings imply a mode of translation control whereby, as an upstream effector of TOR, ROP2 coordinates TOR function in translation reinitiation pathways in response to auxin. © 2017 The Authors.

A tale of five cities: Using recycling frameworks to analyse inclusive recycling performance.

Science.gov (United States)

Scheinberg, Anne; Simpson, Michael

2015-11-01

'Recycling' is a source of much confusion, particularly when comparing solid waste systems in high-income countries with those in low- and middle-income countries. Few analysts can explain why the performance and structure of recycling appears to be so different in rich countries from poor ones, nor why well-meaning efforts to implement recycling so often fail. The analysis of policy drivers, and the Integrated Sustainable Waste Management (ISWM) framework, come close to an explanation.This article builds on these earlier works, focusing in on five cities profiled in the 2010 UN-Habitat publication (Scheinberg A, Wilson DC and Rodic L (2010) Solid Waste Management in the World's Cities. UN-Habitat's Third Global Report on the State of Water and Sanitation in the World's Cities. Newcastle-on-Tyne, UK: Earthscan Publications). Data from these cities and others provides the basis for developing a new tool to analyse inclusive recycling performance. The points of departure are the institutional and economic relationships between the service chain, the public obligation to remove waste, pollution, and other forms of disvalue, and the value chain, a system of private enterprises trading valuable materials and providing markets for recyclables. The methodological innovation is to use flows of materials and money as indicators of institutional relationships, and is an extension of process flow diagramming.The authors are using the term 'recycling framework analysis' to describe this new form of institutional analysis. The diagrams increase our understanding of the factors that contribute to high-performance inclusive recycling. By focusing on institutional relationships, the article seeks to improve analysis, planning, and ultimately, outcomes, of recycling interventions. © The Author(s) 2015.

Resource Efficient Metal and Material Recycling

Science.gov (United States)

Reuter, Markus A.; van Schaik, Antoinette

Metals enable sustainability through their use and their recyclability. However, various factors can affect the Resource Efficiency of Metal Processing and Recycling. Some typical factors that enable Resource Efficiency include and arranged under the drivers of sustainability: Environment (Maximize Resource Efficiency — Energy, Recyclates, Materials, Water, Sludges, Emissions, Land); Economic Feasibility (BAT & Recycling Systems Simulation / Digitalization, Product vis-à-vis Material Centric Recycling); and Social — Licence to Operate (Legislation, consumer, policy, theft, manual labour.). In order to realize this primary production has to be linked systemically with typical actors in the recycling chain such as Original Equipment Manufacturers (OEMs), Recyclers & Collection, Physical separation specialists as well as process metallurgical operations that produce high value metals, compounds and products that recycle back to products. This is best done with deep knowledge of multi-physics, technology, product & system design, process control, market, life cycle management, policy, to name a few. The combination of these will be discussed as Design for Sustainability (DfS) and Design for Recycling (DfR) applications.

Printability of papers recycled from toner and inkjet-printed papers after deinking and recycling processes.

Science.gov (United States)

Karademir, Arif; Aydemir, Cem; Tutak, Dogan; Aravamuthan, Raja

2018-04-01

In our contemporary world, while part of the fibers used in the paper industry is obtained from primary fibers such as wood and agricultural plants, the rest is obtained from secondary fibers from waste papers. To manufacture paper with high optical quality from fibers of recycled waste papers, these papers require deinking and bleaching of fibers at desired levels. High efficiency in removal of ink from paper mass during recycling, and hence deinkability, are especially crucial for the optical and printability quality of the ultimate manufactured paper. In the present study, deinkability and printability performance of digitally printed paper with toner or inkjet ink were compared for the postrecycling product. To that end, opaque 80 g/m 2 office paper was digitally printed under standard printing conditions with laser toner or inkjet ink; then these sheets of paper were deinked by a deinking process based on the INGEDE method 11 p. After the deinking operation, the optical properties of the obtained recycled handsheets were compared with unprinted (reference) paper. Then the recycled paper was printed on once again under the same conditions as before with inkjet and laser printers, to monitor and measure printing color change before and after recycling, and differences in color universe. Recycling and printing performances of water-based inkjet and toner-based laser printed paper were obtained. The outcomes for laser-printed recycled paper were better than those for inkjet-printed recycled paper. Compared for luminosity Y, brightness, CIE a* and CIE b* values, paper recycled from laser-printed paper exhibited higher value than paper recycled from inkjet-printed paper.

Recycling of Glass

DEFF Research Database (Denmark)

Christensen, Thomas Højlund; Damgaard, Anders

2011-01-01

Glass is used for many purposes, but in the waste system glass is predominantly found in terms of beverage and food containers with a relatively short lifetime before ending up in the waste. Furthermore there is a large amount of flat glass used in building materials which also ends up in the waste...... system; this glass though has a long lifetime before ending up in the waste. Altogether these product types add up to 82% of the production of the European glass industry (IPCC, 2001). Recycling of glass in terms of cleaning and refilling of bottles as well as the use of broken glass in the production...... of new glass containers is well established in the glass industry. This chapter describes briefly howglass is produced and howwaste glass is recycled in the industry. Quality requirements and use of recycled products are discussed, as are the resource and environmental issues of glass recycling....

Study of Aramid Fiber/Polychloroprene Recycling Process by Thermal Degradation

Directory of Open Access Journals (Sweden)

Igor Dabkiewicz

2016-07-01

Full Text Available Aramid fiber is an important polymer applied as reinforcement in high-performance composites, which, due its exceptional properties, becomes an excellent impact absorption material. It has been broadly utilized in aeronautic industry and ballistic protection. In aircrafts, it is mainly used in secondary structures, such as fairings, floor panels, and bullet proof structures in helicopters, whereas, in ballistic protection industry, it is applied in automotive armor and bullet proof vest. Under environmental perspective, it is worrying the development and application of composites, which generate proportional discards of these materials, whether originated from manufacturing process, spare parts or end of life cycle. High-performance composite materials like those using aramid fiber are generally difficult to recycle due to their properties and the difficulty for the separation of the components, making their recycling economically unviable. From the characteristics of composite materials and environmental viewpoint, this paper presents a new aramid fiber recycling process. The main objective of this research was to study different recycling methods in aramid fibers/Neoprene® composites. To promote the Neoprene® degradation, it was used a pyrolysis oven with controlled atmosphere and CO 2 injection. For the degraded separation, it was designed a mechanical washing machine in which the most degraded separation occurred. To complete the materials separation, it was employed a manual cleaning process, and, at least to prove the efficacy of the process, it was applied a tensile test in the yarns.

You're a "What"? Recycling Coordinator

Science.gov (United States)

Torpey, Elka Maria

2011-01-01

Recycling coordinators supervise curbside and dropoff recycling programs for municipal governments or private firms. Today, recycling is mandatory in many communities. And advancements in collection and processing methods have helped to increase the quantity of materials for which the recycling coordinator is responsible. In some communities,…

WATER RESISTANCE OF RECYCLED PAPER PANEL

Directory of Open Access Journals (Sweden)

Alexander Rani Suryandono

2017-06-01

Alice Wisler (2015 Facts about Recycling Paper. http://greenliving.lovetoknow.com/Facts_About_Recycling_Paper. Accessed 2 April 2016 Clay Miller (2011 5 Benefits of Recycling Paper. http://www.ways2gogreenblog.com/2011/09/28/5-benefits-of-recycling-paper/. Accessed 10 May 2016 Hari Goyal (2015 Grades of Paper. http://www.paperonweb.com/grade.htm. Accessed 2 April 2016 Hari Goyal (2015 Properties of Paper. http://www.paperonweb.com/paperpro.htm. Accessed 2 April 2016 Kathryn Sukalich (2016 Everything You Need to Know about Paper Recycling. http://earth911.com/business-policy/business/paper-recycling-details-basics/. Accessed 15 July 2016 [U1] Larry West (2015 Why Recycle Paper. http://environment.about.com/od/recycling/a/The-Benefits-Of-Paper-Recycling-Why-Recycle-Paper.htm. Accesed 15 June 2016 Marie-Luise Blue (2008 The Advantages of Recycling Paper. http://education.seattlepi.com/advantages-recycling-paper-3440.html. Accessed 15 June 2016 Nina Spitzer (2009 http://www.sheknows.com/home-and-gardening/articles/810025/the-impact-of-disposable-coffee-cups-on-the-environment. Accessed 15 June 2016 Radio New Zealand (2010 Iwi not Giving Up Fight against Tasman Mill Discharges. http://www.radionz.co.nz/news/regional/64521/iwi-not-giving-up-fight-against-tasman-mill-discharges. Accessed 15 July 2016 Rick LeBlanc (2016 Paper Recycling Facts, Figures and Information Sources. https://www.thebalance.com/paper-recycling-facts-figures-and-information-sources-2877868?_ga=1.192832942.544061388.1477446686. Accesed 2 April 2016 Robinson Meyer (2016 Will More Newspapers Go Nonprofit? http://www.theatlantic.com/technology/archive/2016/01/newspapers-philadelphia-inquirer-daily-news-nonprofit-lol-taxes/423960/. Accessed 3 August 2016 School of Engineering at Darthmouth (2010 Forest and Paper Industry. http://engineering.dartmouth.edu/~d30345d/courses/engs171/Paper.pdf. Accessed 2 April 2016 T. Subramani, V. Angappan. (2015. Experimental Investigation of Papercrete Concrete

Technology options for future recycling

International Nuclear Information System (INIS)

Kikuchi, T.

2001-01-01

Recycling of nuclear material is indispensable, not only for using valuable resources but also for reducing the debt which we may leave to the next generations. Advanced reprocessing technologies have been developed in several countries to deal with the diversification of nuclear fuels. Also technologies derived from reprocessing or other fuel cycle areas have continued to be developed in terms of recycling. Cost effectiveness and waste-free processing are increasingly important factors in the applicable of an alternate recycling policy. This paper introduces an example of the studies in this field conducted in some countries including Japan and considers the establishment of effective recycling methodologies taking into account the uncertainty of future recycling policy. (author)

Rab22a enhances CD147 recycling and is required for lung cancer cell migration and invasion.

Science.gov (United States)

Zhou, Yang; Wu, Bo; Li, Jiang-Hua; Nan, Gang; Jiang, Jian-Li; Chen, Zhi-Nan

2017-08-01

Rab22a is a member of the Ras-related small GTPase family, which plays a key role in regulating the recycling of cargo proteins entering cells through clathrin-independent endocytosis (CIE). Rab22a is overexpressed in different cancer types, including liver cancer, malignant melanoma, ovarian cancer and osteosarcoma. However, its oncogenic role remains unknown. In this study, we found that silencing of Rab22a suppressed the migration and invasion of lung cancer cells. Furthermore, Rab22a interacts with CD147, and knockdown of Rab22a blocks CD147 recycling and promotes CD147 degradation. Taken together, our findings indicate that Rab22a enhances recycling of CD147, which is required for lung cancer cell migration and invasion,and targeting CD147 recycling may be a rational strategy for lung cancer therapy. Copyright © 2017. Published by Elsevier Inc.

The nonlinear relationship between paper recycling and primary pulp requirements : modeling paper production and recycling in Europe

NARCIS (Netherlands)

Schenk, Niels J.; Moll, Henri C.; Potting, Josepha

Waste paper is suitable for recycling back into paper or for incineration for energy recovery. If waste paper is used for recycling, secondary pulp replaces virgin pulp. Fiber recycling is limited, however, because of physical constraints—particularly the breakage of fiber in the recycling

The Three Rs: Reduce, Reuse, Recycle.

Science.gov (United States)

Science Activities, 1991

1991-01-01

A student hand-out for a recycling unit defines the terms reduce, recycle, and reuse as they relate to solid waste management. Presents the characteristics of recyclable items such as yard wastes, metals, glass, and paper. Lists organizations through which more information about recycling can be obtained. (MCO)

Reuse, Reduce, Recycle.

Science.gov (United States)

Briscoe, Georgia

1991-01-01

Discussion of recycling paper in law libraries is also applicable to other types of libraries. Results of surveys of law libraries that investigated recycling practices in 1987 and again in 1990 are reported, and suggestions for reducing the amount of paper used and reusing as much as possible are offered. (LRW)

Cellular regulation of the dopamine transporter

DEFF Research Database (Denmark)

Eriksen, Jacob

2010-01-01

-membrane spanning protein Tac, thereby creating an extracellular antibody epitope. Upon expression in HEK293 cells this TacDAT fusion protein displayed functional properties similar to the wild type transporter. In an ELISA based internalization assay, TacDAT intracellular accumulation was increased by inhibitors...... of lysosomal degradation and moreover TacDAT colocalized with the late endosomal marker Rab7. In the DA cell line 1Rb3An27 TacDAT also co-localized with EGFP-Rab7 and not with the recycling endosomal marker EGFP-Rab11. To assess whether sorting to late endosomes/lysosomes was a property also inherent...... to natively expressed transporter, DAT was visualized directly in cultured DA neurons using the fluorescent cocaine analog JHC 1-64. These data showed pronounced colocalization upon constitutive internalization with Lysotracker, a late endosomal/lysosomal marker; however only little cololization was observed...

What can recycling in thermal reactors accomplish?

International Nuclear Information System (INIS)

Piet, Steven J.; Matthern, Gretchen E.; Jacobson, Jacob J.

2007-01-01

Thermal recycle provides several potential benefits when used as stop-gap, mixed, or backup recycling to recycling in fast reactors. These three roles involve a mixture of thermal and fast recycling; fast reactors are required to some degree at some time. Stop-gap uses thermal reactors only until fast reactors are adequately deployed and until any thermal-recycle-only facilities have met their economic lifetime. Mixed uses thermal and fast reactors symbiotically for an extended period of time. Backup uses thermal reactors only if problems later develop in the fast reactor portion of a recycling system. Thermal recycle can also provide benefits when used as pure thermal recycling, with no intention to use fast reactors. However, long term, the pure thermal recycling approach is inadequate to meet several objectives. (authors)

What can Recycling in Thermal Reactors Accomplish?

International Nuclear Information System (INIS)

Steven Piet; Gretchen E. Matthern; Jacob J. Jacobson

2007-01-01

Thermal recycle provides several potential benefits when used as stop-gap, mixed, or backup recycling to recycling in fast reactors. These three roles involve a mixture of thermal and fast recycling; fast reactors are required to some degree at some time. Stop-gap uses thermal reactors only until fast reactors are adequately deployed and until any thermal-recycle-only facilities have met their economic lifetime. Mixed uses thermal and fast reactors symbiotically for an extended period of time. Backup uses thermal reactors only if problems later develop in the fast reactor portion of a recycling system. Thermal recycle can also provide benefits when used as pure thermal recycling, with no intention to use fast reactors. However, long term, the pure thermal recycling approach is inadequate to meet several objectives

Implementing a campus wide recycling program

International Nuclear Information System (INIS)

Alvarez, L.

2002-01-01

'Full text:' The University of Windsor is currently expanding its recycling program to include all buildings on campus, but faces two challenges: 1) uncertainty about the current waste composition and distribution on campus; and 2) uncertainty about the effectiveness of increased recycling. This project assesses the current waste composition and the attitudes of the students towards recycling, and evaluates the effectiveness of proposed recycling activities. At present, paper is the only material that is collected throughout the entire campus. Except for two buildings, all other potentially recyclable materials within buildings, such as metal, glass, and plastic beverage containers, are discarded. The main focus of this research is on beverage containers as they represent clearly identifiable materials, but other materials were examined as well. To quantify the waste, different buildings on campus were classified according to their function: academic,operational and administrative. The waste composition study indicated that approximately 33% of the campus waste which is landfilled is composed of potentially recyclable material. A survey was then conducted to gauge the campus population's views on recycling issues that could affect the design of a recycling program. Interestingly, 97% of the respondents indicated a high willingness to recycle, but were uncertain as to how and where to recycle on campus. The project is currently assessing potential diversion rates using new, clearly identifiable recycling receptacles placed within selected classrooms for all major materials. There is a significant tradeoff however because the cost for new receptacles is considerable: multiple materials containers are often placed in high pedestrian traffic locations (e.g., hallways) and not always in classrooms,of which there are often many. This project will evaluate the basic benefits and costs of implementing a more comprehensive recycling program, and recommend how other

Lamps recycling aiming at the environment preservation

International Nuclear Information System (INIS)

Yamachita, Roberto Akira; Gama, Paulo Henrique R. Pereira; Haddad, Jamil; Santos, Afonso H. Moreira; Guardia, Eduardo C.

1999-01-01

The article discusses the following issues of lamps recycling in Brazil: mercury lamps recycling, recycling potential, energy conservation and environmental impacts, enterprises lamps recycling, and incentives policy

An industry response to recycle 2000

International Nuclear Information System (INIS)

Motl, G.P.; Loiselle, V.

1996-01-01

The US DOE is expected to issue a policy early this year articulating DOE's position on the recycle of DOE radioactive scrap metal. In anticipation of this 'Recycle 2000' initiative, the nuclear industry has formed a new trade association called the Association of Radioactive Metal Recyclers (ARMR). This article describes the Recycle 2000 initiative, provides some background on the ARMR and its membership, and identifies industry views on the actions to be taken and issues to be resolved in Recycle 2000 is to become a reality

Structure of the LDL receptor extracellular domain at endosomalpH

Energy Technology Data Exchange (ETDEWEB)

Rudenko, Gabby; Henry, Lisa; Henderson, Keith; Ichtchenko,Konstantin; Brown, Michael S.; Goldstein, Joseph L.; Deisenhofer, Johann

2002-09-05

The structure of the low-density lipoprotein receptor extracellular portion has been determined. The document proposes a mechanism for the release of lipoprotein in the endosome. Without this release, the mechanism of receptor recycling cannot function.

Recycling Behavior: A Multidimensional Approach

Science.gov (United States)

Meneses, Gonzalo Diaz; Palacio, Asuncion Beerli

2005-01-01

This work centers on the study of consumer recycling roles to examine the sociodemographic and psychographic profile of the distribution of recycling tasks and roles within the household. With this aim in mind, an empirical work was carried out, the results of which suggest that recycling behavior is multidimensional and comprises the undertaking…

Attributes to facilitate e-waste recycling behaviour

Directory of Open Access Journals (Sweden)

Senawi Nur Hidayah

2016-01-01

Full Text Available This study aims to identify the set of attributes to facilitate electronic waste (e-waste behaviour among the community. E-waste disposal is increasing from year to year in parallel with increasing of global population. The short lifespan of electronics and poor e-waste recycling behaviour is among the main contributors to the steadily increasing of e-waste generated. Current recycling rate among the nation is lacking behind, which is only 10.5%. A questionnaire survey has been conducted among the students in Universiti Teknologi Malaysia to evaluate the current e-waste recycling practice. The results showed that majority of the respondents did not recycle their e-waste on campus. Aggressive efforts is needed to realize the country’s target of 20% recycling rate in year 2020, one of the effective paths is to minimize e-waste generation via active e-waste recycling behaviour among the community. Extensive literatures have been reviewed to classify the attributes to facilitate effective e-waste recycling among the community. Total of five attributes that identified in this study which are Convenience of E- waste Recycling Infrastruture and Services, E-waste Recycling Information, Incentives For E-waste Recycling, Reminder to Recycle E-waste And E-waste Recycling Infrastructure and Services. The set of attributes identified in this study may serve as guideline for the management in designing program to foster e-waste recycling behaviour among the community.

Endosomal protein sorting and autophagy genes contribute to the regulation of yeast life span.

Science.gov (United States)

Longo, Valter D; Nislow, Corey; Fabrizio, Paola

2010-11-01

Accumulating evidence from various organisms points to a role for autophagy in the regulation of life span. By performing a genome-wide screen to identify novel life span determinants in Saccharomyces cerevisiae, we have obtained further insights into the autophagy-related and -unrelated degradation processes that may be important for preventing cellular senescence. The generation of multivesicular bodies and their fusion with the vacuole in the endosomal pathway emerged as novel cell functions involved in yeast chronological survival and longevity extension.

Recycling of used oil

International Nuclear Information System (INIS)

Vipulanandan, C.; Ghurye, G.

1992-01-01

This paper reports on used oil which is a valuable resource that should be recycled. Recycling used oil saves energy and natural resources. Used oil can be reprocessed and used as fuel in industrial burners and boilers. Unfortunately, more than 400 million gallons/year of used oil is lost through widespread dumping, partly due to lack of effective recycling procedures. Although used oil is not currently a federally listed hazardous waste, the U.S. EPA has proposed to list it as a hazardous waste, which will make recycling of used oil even more attractive. Laboratory samples, representing used oil, were used for detailed parametric studies and to determine the limitation of extending some of the current physical separation techniques such as sedimentation and centrifuging developed for oil-water and solid-liquid separation

Challenges in plastics recycling

DEFF Research Database (Denmark)

Pivnenko, Kostyantyn; Jakobsen, L. G.; Eriksen, Marie Kampmann

2015-01-01

Recycling of waste plastics still remains a challenging area in the waste management sector. The current and potential goals proposed on EU or regional levels are difficult to achieve, and even to partially fullfil them the improvements in collection and sorting should be considerable. A study...... was undertaken to investigate the factors affecting quality in plastics recycling. The preliminary results showed factors primarily influencing quality of plastics recycling to be polymer cross contamination, presence of additives, non-polymer impurities, and polymer degradation. Deprivation of plastics quality......, with respect to recycling, has been shown to happen throughout the plastics value chain, but steps where improvements may happen have been preliminary identified. Example of Cr in plastic samples analysed showed potential spreading and accumulation of chemicals ending up in the waste plastics. In order...

Studies on recycled aggregates-based concrete.

Science.gov (United States)

Rakshvir, Major; Barai, Sudhirkumar V

2006-06-01

Reduced extraction of raw materials, reduced transportation cost, improved profits, reduced environmental impact and fast-depleting reserves of conventional natural aggregates has necessitated the use of recycling, in order to be able to conserve conventional natural aggregate. In this study various physical and mechanical properties of recycled concrete aggregates were examined. Recycled concrete aggregates are different from natural aggregates and concrete made from them has specific properties. The percentages of recycled concrete aggregates were varied and it was observed that properties such as compressive strength showed a decrease of up to 10% as the percentage of recycled concrete aggregates increased. Water absorption of recycled aggregates was found to be greater than natural aggregates, and this needs to be compensated during mix design.

Innovative Vacuum Distillation for Magnesium Recycling

Science.gov (United States)

Zhu, Tianbai; Li, Naiyi; Mei, Xiaoming; Yu, Alfred; Shang, Shixiang

Magnesium recycling now becomes a very important subject as magnesium consumption increases fast around the world. All commonly used magnesium die-casting alloys can be recycled and recovered to the primary metal quality. The recycled materials may be comprised of biscuits, sprues, runners, flash, overflows, dross, sludge, scrap parts, and old parts that are returned from service, An innovative magnesium recycle method, vacuum distillation, is developed and proved out to be able to recycle magnesium scraps, especially machining chips, oily magnesium, smelting sludge, dross or the mixture. With this process at a specific temperature and environment condition, magnesium in scraps can be gasified and then solidified to become crystal magnesium crown. This `recycled' magnesium crown is collected and used as the raw material of magnesium alloys. The experimental results show the vacuum distillation is a feasible and plausible method to recycle magnesium. Further, the cost analysis will be addressed in this paper.

Management and recycling of electronic waste

International Nuclear Information System (INIS)

Tanskanen, Pia

2013-01-01

Waste electrical and electronic equipment (WEEE) is one of the largest growing waste streams globally. Hence, for a sustainable environment and the economic recovery of valuable material for reuse, the efficient recycling of electronic scrap has been rendered indispensable, and must still be regarded as a major challenge for today’s society. In contrast to the well-established recycling of metallic scrap, it is much more complicated to recycle electronics products which have reached the end of their life as they contain many different types of material types integrated into each other. As illustrated primarily for the recycling of mobile phones, the efficient recycling of WEEE is not only a challenge for the recycling industry; it is also often a question of as-yet insufficient collection infrastructures and poor collection efficiencies, and a considerable lack of the consumer’s awareness for the potential of recycling electronics for the benefit of the environment, as well as for savings in energy and raw materials

Investigation about the ecotown-enterprise for establishing recycling system of non-radioactive waste arising from power plant decommissioning

International Nuclear Information System (INIS)

Hironaga, Michihiko; Nishiuchi, Tatsuo; Ozaki, Yukio; Yamamoto, Kimio

2004-01-01

About 95% of demolition wastes generated by decommissioning nuclear power plants are below the clearance level, i.e., the wastes can be dealt with as industrial wastes. On that case, rational processing, disposal, and reuse are expectable. However, even if the demolition waste is below a clearance level, it seems to be difficult to be immediately accepted in general society with the demolition wastes. Therefore, it is important to establish the technology for an effective recycle system of demolition wastes, and to reuse demolition wastes as much as possible, resulting in recognition of the value by the society. On the other hand, as for recycling of industrial waste, the recycling enterprise is promoted in the domestic self-governing body in response to the 'eco-town enterprise' which is recommended by the government. This report investigates the system and subjects of a 'eco-town enterprise' for recycling demolition wastes. (author)

The Compressor Recycle System

OpenAIRE

Barstad, Bjørn Ove

2010-01-01

The compressor recycle system is the main focus of this thesis. When the mass flow through a compressor becomes too low, the compressor can plunge into surge. Surge is a term that is used for axisymmetric oscillation through a compressor and is highly unwanted. The recycle system feeds compressed gas back to the intake when the mass flow becomes too low, and thereby act as a safety system.A mathematical model of the recycle system is extended and simulated in SIMULINK. The mathematical model ...

Cascade use indicators for selected biopolymers: Are we aiming for the right solutions in the design for recycling of bio-based polymers?

Science.gov (United States)

Hildebrandt, Jakob; Bezama, Alberto; Thrän, Daniela

2017-04-01

When surveying the trends and criteria for the design for recycling (DfR) of bio-based polymers, priorities appear to lie in energy recovery at the end of the product life of durable products, such as bio-based thermosets. Non-durable products made of thermoplastic polymers exhibit good properties for material recycling. The latter commonly enjoy growing material recycling quotas in countries that enforce a landfill ban. Quantitative and qualitative indicators are needed for characterizing progress in the development towards more recycling friendly bio-based polymers. This would enable the deficits in recycling bio-based plastics to be tracked and improved. The aim of this paper is to analyse the trends in the DfR of bio-based polymers and the constraints posed by the recycling infrastructure on plastic polymers from a systems perspective. This analysis produces recommendations on how life cycle assessment indicators can be introduced into the dialogue between designers and recyclers in order to promote DfR principles to enhance the cascading use of bio-based polymers within the bioeconomy, and to meet circular economy goals.

Cascade use indicators for selected biopolymers: Are we aiming for the right solutions in the design for recycling of bio-based polymers?

Science.gov (United States)

Hildebrandt, Jakob; Bezama, Alberto; Thrän, Daniela

2017-01-01

When surveying the trends and criteria for the design for recycling (DfR) of bio-based polymers, priorities appear to lie in energy recovery at the end of the product life of durable products, such as bio-based thermosets. Non-durable products made of thermoplastic polymers exhibit good properties for material recycling. The latter commonly enjoy growing material recycling quotas in countries that enforce a landfill ban. Quantitative and qualitative indicators are needed for characterizing progress in the development towards more recycling friendly bio-based polymers. This would enable the deficits in recycling bio-based plastics to be tracked and improved. The aim of this paper is to analyse the trends in the DfR of bio-based polymers and the constraints posed by the recycling infrastructure on plastic polymers from a systems perspective. This analysis produces recommendations on how life cycle assessment indicators can be introduced into the dialogue between designers and recyclers in order to promote DfR principles to enhance the cascading use of bio-based polymers within the bioeconomy, and to meet circular economy goals. PMID:28097922

Use of recycled fine aggregate in concretes with durable requirements.

Science.gov (United States)

Zega, Claudio Javier; Di Maio, Angel Antonio

2011-11-01

The use of construction waste materials as aggregates for concrete production is highly attractive compared to the use of non-renewable natural resources, promoting environmental protection and allowing the development of a new raw material. Several countries have recommendations for the use of recycled coarse aggregate in structural concrete, whereas the use of the fine fraction is limited because it may produce significant changes in some properties of concrete. However, during the last decade the use of recycled fine aggregates (RFA) has achieved a great international interest, mainly because of economic implications related to the shortage of natural sands suitable for the production of concrete, besides to allow an integral use of this type of waste. In this study, the durable behaviour of structural concretes made with different percentage of RFA (0%, 20%, and 30%) is evaluated. Different properties related to the durability of concretes such as absorption, sorptivity, water penetration under pressure, and carbonation are determined. In addition, the results of compressive strength, static modulus of elasticity and drying shrinkage are presented. The obtained results indicate that the recycled concretes have a suitable resistant and durable behaviour, according to the limits indicated by different international codes for structural concrete. Copyright © 2011 Elsevier Ltd. All rights reserved.

MOX recycling-an industrial reality

International Nuclear Information System (INIS)

Shallo, G.D.F.

1996-01-01

Reprocessing and plutonium recycling have now attained industrial maturity in France and Europe. Specifically, mixed-oxide (MOX) fuel is fabricated and used in light water reactors (LWRs) in satisfactory operating conditions. The utilities and the fuel cycle industry experience no technical difficulties, and European recycling programs are growing steadily, from 18 reactors in operation today up to 50 expected around the year 2000, putting the system reprocessing-recycling in coherence: 25 t of plutonium will then be used each year to produce the electricity equivalence of 25 millions tons of oil. Plutonium recycling in MOX fuel in current LWRs proves to be technically safe and economically competitive and meets natural resource savings and environmental protection objectives. And recycling responds properly to the nonproliferation concerns. Such an industrial experience gives a unique reference for weapons plutonium disposition through MOX use in reactors

ZZ WPPR, Pu Recycling Benchmark Results

International Nuclear Information System (INIS)

Lutz, D.; Mattes, M.; Delpech, Marc; Juanola, Marc

2002-01-01

Description of program or function: The NEA NSC Working Party on Physics of Plutonium Recycling has commissioned a series of benchmarks covering: - Plutonium recycling in pressurized-water reactors; - Void reactivity effect in pressurized-water reactors; - Fast Plutonium-burner reactors: beginning of life; - Plutonium recycling in fast reactors; - Multiple recycling in advanced pressurized-water reactors. The results have been published (see references). ZZ-WPPR-1-A/B contains graphs and tables relative to the PWR Mox pin cell benchmark, representing typical fuel for plutonium recycling, one corresponding to a first cycle, the second for a fifth cycle. These computer readable files contain the complete set of results, while the printed report contains only a subset. ZZ-WPPR-2-CYC1 are the results from cycle 1 of the multiple recycling benchmarks

Frequent Questions on Recycling

Science.gov (United States)

This is a list of frequent questions on recycling, broken down into five categories. These are answers to common questions that EPA has received from press and web inquiries. This list is located on the Reduce, Reuse, Recycle website.

Recycled concrete with coarse recycled aggregate. An overview and analysis

Directory of Open Access Journals (Sweden)

B. González-Fonteboa

2018-04-01

Full Text Available The construction field has contributed to environmental degradation, producing a high amount of construction and demolition waste (C&D waste and consuming large volumes of natural resources. In this context, recycled concrete (RC has been recognised as a means to preserve natural resources and reduce space for waste storage. During the last decades, many researchers have developed works studying different recycled concrete properties. This review focuses on structural RC made with coarse recycled aggregate from concrete waste. The main objective is to provide a state of the art report on RC’s properties and an analysis on how to predict them taking into account relevant research works. Moreover, the study tries to collect and update RC findings, proposing equations to define RC’s performance, in terms of mechanical strength, modulus of elasticity, stress-strain, creep and shrinkage.

Solid waste recycling in Rajshahi city of Bangladesh.

Science.gov (United States)

Bari, Q Hamidul; Hassan, K Mahbub; Haque, M Ehsanul

2012-11-01

Efficient recycling of solid wastes is now a global concern for a sustainable and environmentally sound management. In this study, traditional recycling pattern of solid waste was investigated in Rajshahi municipality which is the fourth largest city of Bangladesh. A questionnaire survey had been carried out in various recycle shops during April 2010 to January 2011. There were 140 recycle shops and most of them were located in the vicinity of Stadium market in Rajshahi. About 1906 people were found to be involved in recycling activities of the city. The major fraction of recycled wastes were sent to capital city Dhaka for further manufacture of different new products. Only a small amount of wastes, specially plastics, were processed in local recycle factories to produce small washing pots and bottle caps. Everyday, an estimated 28.13 tons of recycled solid wastes were handled in Rajshahi city area. This recycled portion accounted for 8.25% of the daily total generated wastes (341 ton d(-1)), 54.6% of total recyclable wastes (51.49 ton d(-1)) and 68.29% of readily recyclable wastes (41.19 ton d(-1)). Major recycled materials were found to be iron, glass, plastic, and papers. Only five factories were involved in preliminary processing of recyclable wastes. Collecting and processing secondary materials, manufacturing recycled-content products, and then buying recycled products created a circle or loop that ensured the overall success of recycling and generated a host of financial, environmental, and social returns. Copyright © 2012 Elsevier Ltd. All rights reserved.

Carambola optics for recycling of light.

Science.gov (United States)

Leutz, Ralf; Fu, Ling; Ries, Harald

2006-04-20

Recycling of light allows the luminance (radiance) emitted by a light source to be increased at the cost of reducing the total luminous flux (radiant power). Recycling of light means returning part of the emitted light to the source, where part of it will escape absorption. An optical design that is suitable for multiple and controlled recycling is described. Carambola optics is named for its resemblance to star fruit. Several pairs of mirrors or prisms redirect light repeatedly onto the source, thus achieving multiple transits of the light through the source. This recycled light exits the carambola in the same phase space as light directly emitted and not recycled.

Text recycling: acceptable or misconduct?

Science.gov (United States)

Harriman, Stephanie; Patel, Jigisha

2014-08-16

Text recycling, also referred to as self-plagiarism, is the reproduction of an author's own text from a previous publication in a new publication. Opinions on the acceptability of this practice vary, with some viewing it as acceptable and efficient, and others as misleading and unacceptable. In light of the lack of consensus, journal editors often have difficulty deciding how to act upon the discovery of text recycling. In response to these difficulties, we have created a set of guidelines for journal editors on how to deal with text recycling. In this editorial, we discuss some of the challenges of developing these guidelines, and how authors can avoid undisclosed text recycling.

Structural recycled concrete: utilization of recycled aggregate from construction and demolition wastes

International Nuclear Information System (INIS)

Alaejos Gutierrez, P.; Sanchez de Juan, M.

2015-01-01

This paper aims to present the main results of CEDEX research works concerning the use of recycled aggregates for structural concretes. By way of conclusion, recommendations on the requirements of the recycled aggregates have been established, providing information about the influence of these aggregates on the properties of structural concrete. (Author)

Plastics recycling: challenges and opportunities.

Science.gov (United States)

Hopewell, Jefferson; Dvorak, Robert; Kosior, Edward

2009-07-27

Plastics are inexpensive, lightweight and durable materials, which can readily be moulded into a variety of products that find use in a wide range of applications. As a consequence, the production of plastics has increased markedly over the last 60 years. However, current levels of their usage and disposal generate several environmental problems. Around 4 per cent of world oil and gas production, a non-renewable resource, is used as feedstock for plastics and a further 3-4% is expended to provide energy for their manufacture. A major portion of plastic produced each year is used to make disposable items of packaging or other short-lived products that are discarded within a year of manufacture. These two observations alone indicate that our current use of plastics is not sustainable. In addition, because of the durability of the polymers involved, substantial quantities of discarded end-of-life plastics are accumulating as debris in landfills and in natural habitats worldwide. Recycling is one of the most important actions currently available to reduce these impacts and represents one of the most dynamic areas in the plastics industry today. Recycling provides opportunities to reduce oil usage, carbon dioxide emissions and the quantities of waste requiring disposal. Here, we briefly set recycling into context against other waste-reduction strategies, namely reduction in material use through downgauging or product reuse, the use of alternative biodegradable materials and energy recovery as fuel. While plastics have been recycled since the 1970s, the quantities that are recycled vary geographically, according to plastic type and application. Recycling of packaging materials has seen rapid expansion over the last decades in a number of countries. Advances in technologies and systems for the collection, sorting and reprocessing of recyclable plastics are creating new opportunities for recycling, and with the combined actions of the public, industry and governments it

Plastics recycling: challenges and opportunities

Science.gov (United States)

Hopewell, Jefferson; Dvorak, Robert; Kosior, Edward

2009-01-01

Plastics are inexpensive, lightweight and durable materials, which can readily be moulded into a variety of products that find use in a wide range of applications. As a consequence, the production of plastics has increased markedly over the last 60 years. However, current levels of their usage and disposal generate several environmental problems. Around 4 per cent of world oil and gas production, a non-renewable resource, is used as feedstock for plastics and a further 3–4% is expended to provide energy for their manufacture. A major portion of plastic produced each year is used to make disposable items of packaging or other short-lived products that are discarded within a year of manufacture. These two observations alone indicate that our current use of plastics is not sustainable. In addition, because of the durability of the polymers involved, substantial quantities of discarded end-of-life plastics are accumulating as debris in landfills and in natural habitats worldwide. Recycling is one of the most important actions currently available to reduce these impacts and represents one of the most dynamic areas in the plastics industry today. Recycling provides opportunities to reduce oil usage, carbon dioxide emissions and the quantities of waste requiring disposal. Here, we briefly set recycling into context against other waste-reduction strategies, namely reduction in material use through downgauging or product reuse, the use of alternative biodegradable materials and energy recovery as fuel. While plastics have been recycled since the 1970s, the quantities that are recycled vary geographically, according to plastic type and application. Recycling of packaging materials has seen rapid expansion over the last decades in a number of countries. Advances in technologies and systems for the collection, sorting and reprocessing of recyclable plastics are creating new opportunities for recycling, and with the combined actions of the public, industry and governments it

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

Science.gov (United States)

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

2015-04-01

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

Recycling in the 90's - a shared responsibility

International Nuclear Information System (INIS)

Anon.

1993-01-01

Recycling means different things to different people. To consumers, recycling can mean putting out bottles and cans for curbside collection. To a product maker - a manufacturer of raw materials, fabricator of goods or products, or brand owner - recycling can mean reformulating goods to include recycled materials. To recycling service providers, recycling can mean providing cost-efficient collection services. To public policy makers in all levels of government recycling can mean establishing collection and utilization regulations. For recycling to work successfully, these diverse groups must work together and share responsibility for its success. Also, if recycling is to succeed on a large scale and over the long term, three critical points must be first addressed: These points are: approach, economics, and markets. These points are discussed

Compressive strength improvement for recycled concrete aggregate

Directory of Open Access Journals (Sweden)

Mohammed Dhiyaa

2018-01-01

Full Text Available Increasing amount of construction waste and, concrete remnants, in particular pose a serious problem. Concrete waste exist in large amounts, do not decay and need long time for disintegration. Therefore, in this work old demolished concrete is crashed and recycled to produce recycled concrete aggregate which can be reused in new concrete production. The effect of using recycled aggregate on concrete compressive strength has been experimentally investigated; silica fume admixture also is used to improve recycled concrete aggregate compressive strength. The main parameters in this study are recycled aggregate and silica fume admixture. The percent of recycled aggregate ranged from (0-100 %. While the silica fume ranged from (0-10 %. The experimental results show that the average concrete compressive strength decreases from 30.85 MPa to 17.58 MPa when the recycled aggregate percentage increased from 0% to 100%. While, when silica fume is used the concrete compressive strength increase again to 29.2 MPa for samples with 100% of recycled aggregate.

Cost effectiveness of recycling: A systems model

Energy Technology Data Exchange (ETDEWEB)

Tonjes, David J., E-mail: david.tonjes@stonybrook.edu [Department of Technology and Society, College of Engineering and Applied Sciences, Stony Brook University, Stony Brook, NY 11794-3560 (United States); Waste Reduction and Management Institute, School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY 11794-5000 (United States); Center for Bioenergy Research and Development, Advanced Energy Research and Technology Center, Stony Brook University, 1000 Innovation Rd., Stony Brook, NY 11794-6044 (United States); Mallikarjun, Sreekanth, E-mail: sreekanth.mallikarjun@stonybrook.edu [Department of Technology and Society, College of Engineering and Applied Sciences, Stony Brook University, Stony Brook, NY 11794-3560 (United States)

2013-11-15

Highlights: • Curbside collection of recyclables reduces overall system costs over a range of conditions. • When avoided costs for recyclables are large, even high collection costs are supported. • When avoided costs for recyclables are not great, there are reduced opportunities for savings. • For common waste compositions, maximizing curbside recyclables collection always saves money. - Abstract: Financial analytical models of waste management systems have often found that recycling costs exceed direct benefits, and in order to economically justify recycling activities, externalities such as household expenses or environmental impacts must be invoked. Certain more empirically based studies have also found that recycling is more expensive than disposal. Other work, both through models and surveys, have found differently. Here we present an empirical systems model, largely drawn from a suburban Long Island municipality. The model accounts for changes in distribution of effort as recycling tonnages displace disposal tonnages, and the seven different cases examined all show that curbside collection programs that manage up to between 31% and 37% of the waste stream should result in overall system savings. These savings accrue partially because of assumed cost differences in tip fees for recyclables and disposed wastes, and also because recycling can result in a more efficient, cost-effective collection program. These results imply that increases in recycling are justifiable due to cost-savings alone, not on more difficult to measure factors that may not impact program budgets.

Reusing recycled aggregates in structural concrete

Science.gov (United States)

Kou, Shicong

The utilization of recycled aggregates in concrete can minimize environmental impact and reduce the consumption of natural resources in concrete applications. The aim of this thesis is to provide a scientific basis for the possible use of recycled aggregates in structure concrete by conducting a comprehensive programme of laboratory study to gain a better understanding of the mechanical, microstructure and durability properties of concrete produced with recycled aggregates. The study also explored possible techniques to of improve the properties of recycled aggregate concrete that is produced with high percentages (≧ 50%) of recycled aggregates. These techniques included: (a) using lower water-to-cement ratios in the concrete mix design; (b) using fly ash as a cement replacement or as an additional mineral admixture in the concrete mixes, and (c) precasting recycled aggregate concrete with steam curing regimes. The characteristics of the recycled aggregates produced both from laboratory and a commercially operated pilot construction and demolition (C&D) waste recycling plant were first studied. A mix proportioning procedure was then established to produce six series of concrete mixtures using different percentages of recycled coarse aggregates with and without the use of fly ash. The water-to-cement (binder) ratios of 0.55, 0.50, 0.45 and 0.40 were used. The fresh properties (including slump and bleeding) of recycled aggregate concrete (RAC) were then quantified. The effects of fly ash on the fresh and hardened properties of RAC were then studied and compared with those RAC prepared with no fly ash addition. Furthermore, the effects of steam curing on the hardened properties of RAC were investigated. For micro-structural properties, the interfacial transition zones of the aggregates and the mortar/cement paste were analyzed by SEM and EDX-mapping. Moreover, a detailed set of results on the fracture properties for RAC were obtained. Based on the experimental

A novel chimeric cell-penetrating peptide with membrane-disruptive properties for efficient endosomal escape.

Science.gov (United States)

Salomone, Fabrizio; Cardarelli, Francesco; Di Luca, Mariagrazia; Boccardi, Claudia; Nifosì, Riccardo; Bardi, Giuseppe; Di Bari, Lorenzo; Serresi, Michela; Beltram, Fabio

2012-11-10

Efficient endocytosis into a wide range of target cells and low toxicity make the arginine-rich Tat peptide (Tat(11): YGRKKRRQRRR, residues 47-57 of HIV-1 Tat protein) an excellent transporter for delivery purposes. Unfortunately, molecules taken up by endocytosis undergo endosomal entrapment and possible metabolic degradation. Escape from the endosome is therefore actively researched. In this context, antimicrobial peptides (AMPs) provide viable templates for the design of new membrane-disruptive motifs. In particular the Cecropin-A and Melittin hybrids (CMs) are among the smallest and most effective peptides with membrane-perturbing abilities. Here we present a novel chimeric peptide in which the Tat(11) motif is fused to the CM(18) hybrid (KWKLFKKIGAVLKVLTTG, residues 1-7 of Cecropin-A and 2-12 of Melittin). When administered to cells, CM(18)-Tat(11) combines the two desired functionalities: efficient uptake and destabilization of endocytotic-vesicle membranes. We show that this chimeric peptide effectively increases cargo-molecule cytoplasm availability and allows the subsequent intracellular localization of diverse membrane-impermeable molecules (i.e. Tat(11)-EGFP fusion protein, calcein, dextrans, and plasmidic DNA) with no detectable cytotoxicity. The present results open the way to the rational engineering of "modular" cell-penetrating peptides (CPPs) that combine (i) efficient translocation from the extracellular milieu into vesicles and (ii) efficient release of molecules from vesicles into the cytoplasm. Copyright © 2012 Elsevier B.V. All rights reserved.

The Diffusion Effect of MSW Recycling

OpenAIRE

Yi-Tui Chen; Fu-Chiang Yang; Shih-Heng Yu

2017-01-01

The purpose of this paper is to compare the recycling performance for some waste fractions selected including food waste, bulk waste, paper, metal products, plastics/rubber and glass products and then to develop some directions for the future improvements. The priority of each waste fraction for recycling is also analyzed by using an importance-performance analysis. Traditionally, the recycling rate that is calculated by the ratio of waste recycled to waste collected is used as an indicator t...

Coolant clean-up and recycle systems

International Nuclear Information System (INIS)

Ito, Takao.

1979-01-01

Purpose: To increase the service life of mechanical seals in a shaft sealing device, eliminate leakages and improve the safety by providing a recycle pump for feeding coolants to a coolant clean-up device upon reactor shut-down and adapting the pump treat only low temperature and low pressure coolants. Constitution: The system is adapted to partially take out coolants from the pipeways of a recycling pump upon normal operation and feed them to a clean-up device. Upon reactor shut-down, the recycle pump is stopped and coolants are extracted by the recycle pump for shut-down into the clean-up device. Since the coolants are not fed to the clean-up device by the recycle pump during normal operation as conducted so far, high temperature and high pressure coolants are not directly fed to the recycle pump, thereby enabling to avoid mechanical problems in the pump. (Kamimura, M.)

Japan's fuel recycling policy

International Nuclear Information System (INIS)

Anon.

1991-01-01

The Atomic Energy Commission (AEC) has formulated Japanese nuclear fuel recycling plan for the next 20 years, based on the idea that the supply and demand of plutonium should be balanced mainly through the utilization of plutonium for LWRs. The plan was approved by AEC, and is to be incorporated in the 'Long term program for development and utilization of nuclear energy' up for revision next year. The report on 'Nuclear fuel recycling in Japan' by the committee is characterized by Japanese nuclear fuel recycling plan and the supply-demand situation for plutonium, the principle of the possession of plutonium not more than the demand in conformity with nuclear nonproliferation attitude, and the establishment of a domestic fabrication system of uranium-plutonium mixed oxide fuel. The total plutonium supply up to 2010 is estimated to be about 85 t, on the other hand, the demand will be 80-90 t. The treatment of plutonium is the key to the recycling and utilization of nuclear fuel. By around 2000, the private sector will commercialize the fabrication of the MOX fuel for LWRs at the annual rate of about 100 t. Commitment to nuclear nonproliferation, future nuclear fuel recycling program in Japan, MOX fuel fabrication system in Japan and so on are reported. (K.I.)

African Swine Fever Virus Undergoes Outer Envelope Disruption, Capsid Disassembly and Inner Envelope Fusion before Core Release from Multivesicular Endosomes.

Directory of Open Access Journals (Sweden)

Bruno Hernáez

2016-04-01

Full Text Available African swine fever virus (ASFV is a nucleocytoplasmic large DNA virus (NCLDV that causes a highly lethal disease in domestic pigs. As other NCLDVs, the extracellular form of ASFV possesses a multilayered structure consisting of a genome-containing nucleoid successively wrapped by a thick protein core shell, an inner lipid membrane, an icosahedral protein capsid and an outer lipid envelope. This structural complexity suggests an intricate mechanism of internalization in order to deliver the virus genome into the cytoplasm. By using flow cytometry in combination with pharmacological entry inhibitors, as well as fluorescence and electron microscopy approaches, we have dissected the entry and uncoating pathway used by ASFV to infect the macrophage, its natural host cell. We found that purified extracellular ASFV is internalized by both constitutive macropinocytosis and clathrin-mediated endocytosis. Once inside the cell, ASFV particles move from early endosomes or macropinosomes to late, multivesicular endosomes where they become uncoated. Virus uncoating requires acidic pH and involves the disruption of the outer membrane as well as of the protein capsid. As a consequence, the inner viral membrane becomes exposed and fuses with the limiting endosomal membrane to release the viral core into the cytosol. Interestingly, virus fusion is dependent on virus protein pE248R, a transmembrane polypeptide of the inner envelope that shares sequence similarity with some members of the poxviral entry/fusion complex. Collective evidence supports an entry model for ASFV that might also explain the uncoating of other multienveloped icosahedral NCLDVs.

Design study of advanced nuclear fuel recycle system. Conceptual study of recycle system using molten salt

International Nuclear Information System (INIS)

Kakehi, I.; Shirai, N.; Hatano, M.; Kajitani, M.; Yonezawa, S.; Kawai, T.; Kawamura, F.; Tobe, K.; Takahashi, K.

1996-12-01

For the purpose of developing the future nuclear fuel recycle system, the design study of the advanced nuclear fuel recycle system is being conducted. This report describes intermediate accomplishments in the conceptual system study of the advanced nuclear fuel recycle system. Fundamental concepts of this system is the recycle system using molten salt which intend to break through the conventional concepts of purex and pellet fuel system. Contents of studies in this period are as follows, 1)feasibility study of the process by Cd-cathode for nitride fuel, 2)application study for the molten salt of low melting point (AlCl3+organic salt), 3)research for decladding (advantage of decladding by heat treatment), 4)behavior of FPs in electrorefining (behavior of iodine and volatile FP chlorides, FPs behavior in chlorination), 5)criticality analysis in electrorefiner, 6)drawing of off-gas flow diagram, 7)drawing of process machinery concept (cathode processor, vibration packing), 8)evaluation for the amounts of the high level radioactive wastes, 9)quality of the recycle fuels (FPs contamination of recycle fuel), 10)conceptual study of in-cell handling system, 11)meaning of the advanced nuclear fuel recycle system. The conceptual system study will be completed in describing concepts of the system and discussing issues for the developments. (author)

Economic and policy instrument analyses in support of the scrap tire recycling program in Taiwan.

Science.gov (United States)

Chang, Ni-Bin

2008-02-01

Understanding the cost-effectiveness and the role of economic and policy instruments, such as the combined product tax-recycling subsidy scheme or a tradable permit, for scrap tire recycling has been of crucial importance in a market-oriented environmental management system. Promoting product (tire) stewardship on one hand and improving incentive-based recycling policy on the other hand requires a comprehensive analysis of the interfaces and interactions in the nexus of economic impacts, environmental management, environmental valuation, and cost-benefit analysis. This paper presents an assessment of the interfaces and interactions between the implementation of policy instruments and its associated economic evaluation for sustaining a scrap tire recycling program in Taiwan during the era of the strong economic growth of the late 1990s. It begins with an introduction of the management of the co-evolution between technology metrics of scrap tire recycling and organizational changes for meeting the managerial goals island-wide during the 1990s. The database collected and used for such analysis covers 17 major tire recycling firms and 10 major tire manufacturers at that time. With estimates of scrap tire generation and possible scale of subsidy with respect to differing tire recycling technologies applied, economic analysis eventually leads to identify the associated levels of product tax with respect to various sizes of new tires. It particularly demonstrates a broad perspective of how an integrated econometric and engineering economic analysis can be conducted to assist in implementing policy instruments for scrap tire management. Research findings indicate that different subsidy settings for collection, processing, and end use of scrap tires should be configured to ameliorate the overall managerial effectiveness. Removing the existing boundaries between designated service districts could strengthen the competitiveness of scrap tires recycling industry, helping to

Probiotics promote endocytic allergen degradation in gut epithelial cells

International Nuclear Information System (INIS)

Song, Chun-Hua; Liu, Zhi-Qiang; Huang, Shelly; Zheng, Peng-Yuan; Yang, Ping-Chang

2012-01-01

Highlights: ► Knockdown of A20 compromised the epithelial barrier function. ► The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. ► Antigens transported across A20-deficient HT-29 monolayers conserved antigenicity. ► Probiotic proteins increased the expression of A20 in HT-29 cells. -- Abstract: Background and aims: Epithelial barrier dysfunction plays a critical role in the pathogenesis of allergic diseases; the mechanism is to be further understood. The ubiquitin E3 ligase A20 (A20) plays a role in the endocytic protein degradation in the cells. This study aims to elucidate the role of A20 in the maintenance of gut epithelial barrier function. Methods: Gut epithelial cell line, HT-29 cell, was cultured into monolayers to evaluate the barrier function in transwells. RNA interference was employed to knock down the A20 gene in HT-29 cells to test the role of A20 in the maintenance of epithelial barrier function. Probiotic derived proteins were extracted from the culture supernatants using to enhance the expression of A20 in HT-29 cells. Results: The results showed that the knockdown of A20 compromised the epithelial barrier function in HT-29 monolayers, mainly increased the intracellular permeability. The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Allergens collected from the transwell basal chambers of A20-deficient HT-29 monolayers still conserved functional antigenicity. Treating with probiotic derived proteins increased the expression of A20 in HT-29 cells and promote the barrier function. Conclusion: A20 plays an important role in the maintenance of epithelial barrier function as shown by HT-29 monolayer. Probiotic derived protein increases the expression of A20 and promote the HT-29 monolayer barrier function.

Probiotics promote endocytic allergen degradation in gut epithelial cells

Energy Technology Data Exchange (ETDEWEB)

Song, Chun-Hua [Department of Epidemiology and Biostatistics, College of Public Health, Zhengzhou University, Zhengzhou (China); Liu, Zhi-Qiang [Department of Gastroenterology, The Second Hospital, Zhengzhou University, Zhengzhou (China); Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON (Canada); Huang, Shelly [Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON (Canada); Zheng, Peng-Yuan, E-mail: medp7123@126.com [Department of Gastroenterology, The Second Hospital, Zhengzhou University, Zhengzhou (China); Yang, Ping-Chang, E-mail: yangp@mcmaster.ca [Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON (Canada)

2012-09-14

Highlights: Black-Right-Pointing-Pointer Knockdown of A20 compromised the epithelial barrier function. Black-Right-Pointing-Pointer The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Black-Right-Pointing-Pointer Antigens transported across A20-deficient HT-29 monolayers conserved antigenicity. Black-Right-Pointing-Pointer Probiotic proteins increased the expression of A20 in HT-29 cells. -- Abstract: Background and aims: Epithelial barrier dysfunction plays a critical role in the pathogenesis of allergic diseases; the mechanism is to be further understood. The ubiquitin E3 ligase A20 (A20) plays a role in the endocytic protein degradation in the cells. This study aims to elucidate the role of A20 in the maintenance of gut epithelial barrier function. Methods: Gut epithelial cell line, HT-29 cell, was cultured into monolayers to evaluate the barrier function in transwells. RNA interference was employed to knock down the A20 gene in HT-29 cells to test the role of A20 in the maintenance of epithelial barrier function. Probiotic derived proteins were extracted from the culture supernatants using to enhance the expression of A20 in HT-29 cells. Results: The results showed that the knockdown of A20 compromised the epithelial barrier function in HT-29 monolayers, mainly increased the intracellular permeability. The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Allergens collected from the transwell basal chambers of A20-deficient HT-29 monolayers still conserved functional antigenicity. Treating with probiotic derived proteins increased the expression of A20 in HT-29 cells and promote the barrier function. Conclusion: A20 plays an important role in the maintenance of epithelial barrier function as shown by HT-29 monolayer. Probiotic derived protein increases the expression of A20 and promote the HT-29 monolayer barrier function.

Achievement report in fiscal 2000 on technological development to recycle waste building materials and glasses. Development of waste building material recycling technology (Research and development of recycling technology corresponding to grades of demolished building lumbers); 2000 nendo kenchiku haizai glass nado recycle gijutsu kaihatsu seika hokokusho. Kenchiku haizai recycle gijutsu kaihatsu (kenchiku kaitai mokuzai no hin'i ni taioshita recycle gijutsu no kenkyu kaihatsu)

Energy Technology Data Exchange (ETDEWEB)

NONE

2001-03-01

With an objective to reduce wastes, and promote effective utilization of wood resources, research and development has been made on a demolished building material recycling technology. This paper summarizes the achievements in fiscal 2000. In developing the technology to manufacture high water resistant wood boards, discussions were given on resor type phenolic resin as an adhesive, and on the medium density fiberboard (MDF) being a substitute material for plywood as the wooden board. As a result, a highly water resistant MDF that can clear JIS E0 has been developed. In the research of a technology to enhance durability of wooden boards, the in-liquid roll press method was devised to perform impregnation of chemicals into board raw materials continually and simply, whose device was fabricated on a trial basis. With regard to recycling of medium to low grade wood-based wastes, researches were performed on pulverization of the wastes, fabrication of liquefied woods, and effective utilization of the liquefied woods. Both of a hammer mill and a chip saw crusher fabricated wood powder with nearly uniform grain size regardless of types of the wood-based wastes. Liquefaction of plywood and PB boards required more stringent reaction conditions than liquefaction of such ordinary members as pillar materials and laminated lumbers. (NEDO)

Influence of recycled aggregate quality and proportioning criteria on recycled concrete properties.

Science.gov (United States)

López-Gayarre, F; Serna, P; Domingo-Cabo, A; Serrano-López, M A; López-Colina, C

2009-12-01

This paper presents the results of experimental research using concrete produced by substituting part of the natural coarse aggregates with recycled aggregates from concrete demolition. The influence of the quality of the recycled aggregate (amount of declassified and source of aggregate), the percentage of replacement on the targeted quality of the concrete to be produced (strength and workability) has been evaluated. The granular structure of concrete and replacement criteria were analyzed in this study, factors which have not been analyzed in other studies. The following properties of recycled concretes were analyzed: density, absorption, compressive strength, elastic modulus, amount of occluded air, penetration of water under pressure and splitting tensile strength. A simplified test program was designed to control the costs of the testing while still producing sufficient data to develop reliable conclusions in order to make the number of tests viable whilst guaranteeing the reliability of the conclusions. Several factors were analyzed including the type of aggregate, the percentage of replacement, the type of sieve curve, the declassified content, the strength of concrete and workability of concrete and the replacement criteria. The type of aggregate and the percentage of replacement were the only factors that showed a clear influence on most of the properties. Compressive strength is clearly affected by the quality of recycled aggregates. If the water-cement ratio is kept constant and the loss of workability due to the effect of using recycled aggregate is compensated for with additives, the percentage of replacement of the recycled aggregate will not affect the compressive strength. The elastic modulus is affected by the percentage of replacement. If the percentage of replacement does not exceed 50%, the elastic modulus will only change slightly.

Reorganization of the Endosomal System in Salmonella-Infected Cells: The Ultrastructure of Salmonella-Induced Tubular Compartments

Science.gov (United States)

Krieger, Viktoria; Liebl, David; Zhang, Yuying; Rajashekar, Roopa; Chlanda, Petr; Giesker, Katrin; Chikkaballi, Deepak; Hensel, Michael

2014-01-01

During the intracellular life of Salmonella enterica, a unique membrane-bound compartment termed Salmonella-containing vacuole, or SCV, is formed. By means of translocated effector proteins, intracellular Salmonella also induce the formation of extensive, highly dynamic membrane tubules termed Salmonella-induced filaments or SIF. Here we report the first detailed ultrastructural analyses of the SCV and SIF by electron microscopy (EM), EM tomography and live cell correlative light and electron microscopy (CLEM). We found that a subset of SIF is composed of double membranes that enclose portions of host cell cytosol and cytoskeletal filaments within its inner lumen. Despite some morphological similarities, we found that the formation of SIF double membranes is independent from autophagy and requires the function of the effector proteins SseF and SseG. The lumen of SIF network is accessible to various types of endocytosed material and our CLEM analysis of double membrane SIF demonstrated that fluid phase markers accumulate only between the inner and outer membrane of these structures, a space continual with endosomal lumen. Our work reveals how manipulation of the endosomal membrane system by an intracellular pathogen results in a unique tubular membrane compartmentalization of the host cell, generating a shielded niche permissive for intracellular proliferation of Salmonella. PMID:25254663

Proceedings of the waste recycling workshop

Energy Technology Data Exchange (ETDEWEB)

Bailey, R.E.; Thomas, A.F.; Ries, M.A. [eds.] [Ohio State Univ., Columbus, OH (United States)

1993-12-31

Recorded are seventeen talks from five sessions at the workshop. FERMCO`s recycling program, state of the art recycling technology, and an integrated demonstration of deactivation, decommissioning and decommissioning are presented in the plenary session. In the concrete session, decontamination and recycling are discussed. In the transite session, regulations are considered along with recycling and decontamination. In the metals session, radioactive scrap metals are emphasized. And in the regulatory considerations and liabilities session, DOE and EPA viewpoints are discussed. (GHH)

MOX fuel reprocessing and recycling

International Nuclear Information System (INIS)

Guillet, J.L.

1990-01-01

This paper is devoted to the reprocessing of MOX fuel in UP2-800 plant at La Hague, and to the MOX successive reprocessing and recycling. 1. MOX fuel reprocessing. In a first step, the necessary modifications in UP2-800 to reprocess MOX fuel are set out. Early in the UP2-800 project, actions have been taken to reprocess MOX fuel without penalty. They consist in measures regarding: Dissolution; Radiological shieldings; Nuclear instrumentation; Criticality. 2. Mox successive reprocessing and recycling. The plutonium recycling in the LWR is now a reality and, as said before, the MOX fuel reprocessing is possible in UP2-800 plant at La Hague. The following actions in this field consist in verifying the MOX successive reprocessing and recycling possibilities. After irradiation, the fissile plutonium content of irradiated MOX fuel is decreased and, in this case, the re-use of plutonium in the LWR need an important increase of initial Pu enrichment inconsistent with the Safety reactor constraints. Cogema opted for reprocessing irradiated MOX fuel in dilution with the standard UO2 fuel in appropriate proportions (1 MOX for 4 UO2 fuel for instance) in order to save a fissile plutonium content compatible with MOX successive recycling (at least 3 recyclings) in LWR. (author). 2 figs

40 CFR 141.76 - Recycle provisions.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Recycle provisions. 141.76 Section 141...) NATIONAL PRIMARY DRINKING WATER REGULATIONS Filtration and Disinfection § 141.76 Recycle provisions. (a... recycle spent filter backwash water, thickener supernatant, or liquids from dewatering processes must meet...

Integration of informal recycling sector in Brazil and the case of Sorocaba City.

Science.gov (United States)

Silva de Souza Lima, Nathalia; Mancini, Sandro Donnini

2017-07-01

Catadores are people who collect and sell materials that can be recycled. This activity has been done informally in many countries for years. Recently, a recognition process has begun for the informal recycling sector, with public and private initiatives. In Brazil, catadores started out associating with each other in co-operatives in the 1980s. In 2010, the Solid Waste National Policy was approved, promoting the inclusion of theses co-operatives in the formal waste management system. However, only in 25 out of 5670 Brazilian municipalities have hired co-operatives as Private Service Providers. The integration of the informal sector has contributed with social, economic and environmental benefits; income generation, reduction of poverty and resource preservation are highlights. Although there was a legal progress, there are great challenges for various actors involved. This paper aims to diagnose the informal recycling sector, emphasizing the integration process that has happened in Brazil. For this, a substantial literature review and a case study were conducted, applying the tool 'InteRa' to the case of Sorocaba. The case showed that it is possible to improve the integration of catadores in the formal waste management system. The co-operatives achieve recycling rates of 2%, higher than the official national rate of 1%. However, we estimate that autonomous pickers increase total recycling in Sorocaba to 9%, still short of the 25% target via source segregation. Therefore, continuing the integration process will benefit both the pickers, and also the municipality through savings on landfill costs.

Effect of Recycle Solvent Hydrotreatment on Oil Yield of Direct Coal Liquefaction

Directory of Open Access Journals (Sweden)

Shansong Gao

2015-07-01

Full Text Available Effects of the recycle solvent hydrotreatment on oil yield of direct coal liquefaction were carried out in the 0.18 t/day direct coal liquefaction bench support unit of National Engineering Laboratory for Direct Coal Liquefaction (China. Results showed that the hydrogen-donating ability of the hydrogenated recycle solvent improved and the hydrogen consumption of solvent hydrotreatment was increased by decreasing liquid hourly space velocity (LHSV from 1.5 to 1.0 h−1 and increasing reaction pressure from 13.7 to 19.0 MPa. The hydrogen-donating ability of the hydrogenated recycle solvent was enhanced, thus promoting the oil yield and coal conversion of the liquefaction reaction. The coal conversion and distillates yield of coal liquefaction were increased from 88.74% to 88.82% and from 47.41% to 49.10%, respectively, with the increase in the solvent hydrotreatment pressure from 13.7 to 19.0 MPa. The coal conversion and distillates of coal liquefaction were increased from 88.82% to 89.27% and from 49.10% to 54.49%, respectively, when the LHSV decreased from 1.5 to 1.0 h−1 under the solvent hydrotreatment pressure of 19.0 MPa.

Downcycling versus recycling of construction and demolition waste: Combining LCA and LCC to support sustainable policy making.

Science.gov (United States)

Di Maria, Andrea; Eyckmans, Johan; Van Acker, Karel

2018-05-01

Urgent solutions are needed in Europe to deal with construction and demolition waste (CDW). EU policy has contributed to significantly reducing the amount of CDW going to landfill, but most of the effort has been put in downcycling practices. Therefore, further policies are needed to stimulate high-quality recycling of CDW. The present paper presents a combined life cycle assessment (LCA) and life cycle costing (LCC) methodologies to analyse the environmental and the economic drivers in four alternative CDW end-of-life scenarios in the region of Flanders, in Belgium. The four analysed alternatives are (i) landfilling, (ii) downcycling, (iii) advanced recycling and (iv) recycling after selective demolition. LCA results show that landiflling is the scenario having the highest environmental impacts in terms of person equivalent (PE), followed by downcycling and recycling (-36%) and recycling after selective demolition (-59%). The decrease in environmental impacts is mostly due to the avoided landfilling of CDW and the recovery of materials from selective demolition. LCC results indicate that landfilling is the scenario bearing the highest total economic costs. This is due to the high landfill tax in Flanders. The recycling after selective demolition bears the second highest cost. The increase of high-quality CDW recycling can significantly reduce the overall environmental impact of the system. Implementing a high landfill tax, increasing the gate fee to the recycling plant, and boosting the sales price of recycled aggregates are the most effective drivers to facilitate a transition towards a more sustainable CDW management system. The paper demonstrates that the combined LCA and LCC results can highlight the environmental and economic drivers in CDW management. The results of the combined analysis can help policymakers to promote the aspects contributing to sustainability and to limit the ones creating a barrier. Copyright © 2018 Elsevier Ltd. All rights reserved.

Auditing Operating Room Recycling: A Management Case Report.

Science.gov (United States)

McGain, Forbes; Jarosz, Katherine Maria; Nguyen, Martin Ngoc Hoai Huong; Bates, Samantha; O'Shea, Catherine Jane

2015-08-01

Much waste arises from operating rooms (ORs). We estimated the practical and financial feasibility of an OR recycling program, weighing all waste from 6 ORs in Melbourne, Australia. Over 1 week, 237 operations produced 1265 kg in total: general waste 570 kg (45%), infectious waste 410 kg (32%), and recyclables 285 kg (23%). The achieved recycling had no infectious contamination. The achieved recycling/potential recycling rate was 285 kg/517 kg (55%). The average waste disposal costs were similar for general waste and recycling. OR recycling rates of 20%-25% total waste were achievable without compromising infection control or financial constraints.

Pep3p/Pep5p complex: a putative docking factor at multiple steps of vesicular transport to the vacuole of Saccharomyces cerevisiae.

OpenAIRE

Srivastava, A; Woolford, C A; Jones, E W

2000-01-01

Pep3p and Pep5p are known to be necessary for trafficking of hydrolase precursors to the vacuole and for vacuolar biogenesis. These proteins are present in a hetero-oligomeric complex that mediates transport at the vacuolar membrane. PEP5 interacts genetically with VPS8, implicating Pep5p in the earlier Golgi to endosome step and/or in recycling from the endosome to the Golgi. To understand further the cellular roles of Pep3p and Pep5p, we isolated and characterized a set of pep3 conditional ...

Electric vehicle recycling 2020: Key component power electronics.

Science.gov (United States)

Bulach, Winfried; Schüler, Doris; Sellin, Guido; Elwert, Tobias; Schmid, Dieter; Goldmann, Daniel; Buchert, Matthias; Kammer, Ulrich

2018-04-01

Electromobility will play a key role in order to reach the specified ambitious greenhouse gas reduction targets in the German transport sector of 42% between 1990 and 2030. Subsequently, a significant rise in the sale of electric vehicles (EVs) is to be anticipated in future. The amount of EVs to be recycled will rise correspondingly after a delay. This includes the recyclable power electronics modules which are incorporated in every EV as an important component for energy management. Current recycling methods using car shredders and subsequent post shredder technologies show high recycling rates for the bulk metals but are still associated with high losses of precious and strategic metals such as gold, silver, platinum, palladium and tantalum. For this reason, the project 'Electric vehicle recycling 2020 - key component power electronics' developed an optimised recycling route for recycling power electronics modules from EVs which is also practicable in series production and can be implemented using standardised technology. This 'WEEE recycling route' involves the disassembly of the power electronics from the vehicle and a subsequent recycling in an electronic end-of-life equipment recycling plant. The developed recycling process is economical under the current conditions and raw material prices, even though it involves considerably higher costs than recycling using the car shredder. The life cycle assessment shows basically good results, both for the traditional car shredder route and the developed WEEE recycling route: the latter provides additional benefits from some higher recovery rates and corresponding credits.

RAB-10-Dependent Membrane Transport Is Required for Dendrite Arborization

Science.gov (United States)

Zou, Wei; Yadav, Smita; DeVault, Laura; Jan, Yuh Nung; Sherwood, David R.

2015-01-01

Formation of elaborately branched dendrites is necessary for the proper input and connectivity of many sensory neurons. Previous studies have revealed that dendritic growth relies heavily on ER-to-Golgi transport, Golgi outposts and endocytic recycling. How new membrane and associated cargo is delivered from the secretory and endosomal compartments to sites of active dendritic growth, however, remains unknown. Using a candidate-based genetic screen in C. elegans, we have identified the small GTPase RAB-10 as a key regulator of membrane trafficking during dendrite morphogenesis. Loss of rab-10 severely reduced proximal dendritic arborization in the multi-dendritic PVD neuron. RAB-10 acts cell-autonomously in the PVD neuron and localizes to the Golgi and early endosomes. Loss of function mutations of the exocyst complex components exoc-8 and sec-8, which regulate tethering, docking and fusion of transport vesicles at the plasma membrane, also caused proximal dendritic arborization defects and led to the accumulation of intracellular RAB-10 vesicles. In rab-10 and exoc-8 mutants, the trans-membrane proteins DMA-1 and HPO-30, which promote PVD dendrite stabilization and branching, no longer localized strongly to the proximal dendritic membranes and instead were sequestered within intracellular vesicles. Together these results suggest a crucial role for the Rab10 GTPase and the exocyst complex in controlling membrane transport from the secretory and/or endosomal compartments that is required for dendritic growth. PMID:26394140

Feedwater recycling system in BWR type reactor

International Nuclear Information System (INIS)

Shimamoto, Yoshiharu.

1980-01-01

Purpose: To improve the reactor safety by preventing thermal stresses and cracks generated in structural materials due to the fluctuations in the temperature for high temperature water - low temperature water mixture near the feedwater nozzle. Method: Feedwater pipes are connected to a pressure vessel not directly but by way of a flow control valve. While the recycled water is circulated from an inlet nozzle to an outlet nozzle through a recycle pump, flow control valve and recycling pipeways, feedwater is fed from the feedwater pipes to the recycling pipeways by way of the flow control valve. More specifically, since the high temperature recycle water and the low temperature recycle water are mixed within the pipeways, the temperature fluctuations resulted from the temperature difference between the recycle water and the feedwater is reduced to prevent thermal fatigue and generation of cracks thereby securing the reactor safety. (Furukawa, Y.)

Technology options for future recycling

International Nuclear Information System (INIS)

Kikuchi, T.

2000-01-01

It goes without saying that recycling of nuclear material is indispensable, not only for the effective use of valuable resources but also to reduce the debt which we may leave to the next generations. Many developments in advanced reprocessing technologies have been carried out in several countries to deal with the diversification of nuclear fuels. Also technologies derived from reprocessing or other fuel cycle areas have continued to be developed in terms of recycling. Cost effectiveness and waste-free processing are increasingly important factors in the applicable of an alternate recycling policy. This paper introduces an example of the studies in this field, which has been conducted in Japan and considers the establishment of effective recycling methodologies taking into account the uncertainty of future policy. (authors)

Management options for recycling radioactive scrap metals

Energy Technology Data Exchange (ETDEWEB)

Dehmel, J.C.; MacKinney, J.; Bartlett, J.

1997-02-01

The feasibility and advantages of recycling radioactive scrap metals (RSM) have yet to be assessed, given the unique technical, regulatory, safety, and cost-benefit issues that have already been raised by a concerned recycling industry. As is known, this industry has been repeatedly involved with the accidental recycling of radioactive sources and, in some cases, with costly consequences. If recycling were deemed to be a viable option, it might have to be implemented with regulatory monitoring and controls. Its implementation may have to consider various and complex issues and address the requirements and concerns of distinctly different industries. There are three basic options for the recycling of such scraps. They are: (1) recycling through the existing network of metal-scrap dealers and brokers, (2) recycling directly and only with specific steelmills, or (3) recycling through regional processing centers. Under the first option, scrap dealers and brokers would receive material from RSM generators and determine at which steelmills such scraps would be recycled. For the second option, RSM generators would deal directly with selected steelmills under specific agreements. For the third option, generators would ship scraps only to regional centers for processing and shipment to participating steelmills. This paper addresses the potential advantages of each option, identifies the types of arrangements that would need to be secured among all parties, and attempts to assess the receptivity of the recycling industry to each option.

Strategies for the Activation and Release of the Membranolytic Peptide Melittin from Liposomes Using Endosomal pH as a Trigger

NARCIS (Netherlands)

Oude Blenke, E.; Sleszynska, M.; Evers, M. J W; Storm, G.; Martin, N. I.; Mastrobattista, E.

2017-01-01

Endosomolytic peptides are often coupled to drug delivery systems to enhance endosomal escape, which is crucial for the delivery of macromolecular drugs that are vulnerable to degradation in the endolysosomal pathway. Melittin is a 26 amino acid peptide derived from bee venom that has a very high

COMMD1 is linked to the WASH complex and regulates endosomal trafficking of the copper transporter ATP7A

NARCIS (Netherlands)

Phillips-Krawczak, Christine A.; Singla, Amika; Starokadomskyy, Petro; Deng, Zhihui; Osborne, Douglas G.; Li, Haiying; Dick, Christopher J.; Gomez, Timothy S.; Koenecke, Megan; Zhang, Jin-San; Dai, Haiming; Sifuentes-Dominguez, Luis F.; Geng, Linda N.; Kaufmann, Scott H.; Hein, Marco Y.; Wallis, Mathew; McGaughran, Julie; Gecz, Jozef; De Sluis, Bart van; Billadeau, Daniel D.; Burstein, Ezra

2015-01-01

COMMD1 deficiency results in defective copper homeostasis, but the mechanism for this has remained elusive. Here we report that COMMD1 is directly linked to early endosomes through its interaction with a protein complex containing CCDC22, CCDC93, and C16orf62. This COMMD/CCDC22/CCDC93 (CCC) complex

Recycling of reprocessed uranium

International Nuclear Information System (INIS)

Randl, R.P.

1987-01-01

Since nuclear power was first exploited in the Federal Republic of Germany, the philosophy underlying the strategy of the nuclear fuel cycle has been to make optimum use of the resource potential of recovered uranium and plutonium within a closed fuel cycle. Apart from the weighty argument of reprocessing being an important step in the treatment and disposal of radioactive wastes, permitting their optimum ecological conditioning after the reprocessing step and subsequent storage underground, another argument that, no doubt, carried weight was the possibility of reducing the demand of power plants for natural uranium. In recent years, strategies of recycling have emerged for reprocessed uranium. If that energy potential, too, is to be exploited by thermal recycling, it is appropriate to choose a slightly different method of recycling from the one for plutonium. While the first generation of reprocessed uranium fuel recycled in the reactor cuts down natural uranium requirement by some 15%, the recycling of a second generation of reprocessed, once more enriched uranium fuel helps only to save a further three per cent of natural uranium. Uranium of the second generation already carries uranium-232 isotope, causing production disturbances, and uranium-236 isotope, causing disturbances of the neutron balance in the reactor, in such amounts as to make further fabrication of uranium fuel elements inexpedient, even after mixing with natural uranium feed. (orig./UA) [de

Synthesis and chemical recycling of high polymers using C1 compounds; C1 kagobutsu ni yoru kobunshi no chemical recycle

Energy Technology Data Exchange (ETDEWEB)

Masuda, T. [National Institute of Materials and Chemical Research, Tsukuba (Japan)

1997-09-01

The paper outlined a study of the synthesis of high polymers using C1 compounds which are continuously usable chemical materials and the related compounds such as the derivatives, and also the chemical recycle. In the case of waste plastics mixed in urban refuse, effective is the chemical recycle where C1 compounds obtained by gasifying the mixed waste are used as high polymer material. For the synthesis and recycle of high polymers using C1 compounds, there are three routes: Route A (recycle via high polymer materials), Route B (recycle via C1 compounds and high polymer materials), and Route C including global-scale carbon recycle (recycle via carbon dioxide from biodegradable plastics using microorganism). Among high polymers, those that can be synthesized from C1 compounds, for example, polymethylene, polyacetal and polyketone can be chemically recycled by Route B. 30 refs., 2 figs., 1 tab.

U.S. Department of Energy National Center of Excellence for Metals Recycle

International Nuclear Information System (INIS)

Adams, V.; Bennett, M.; Bishop, L.

1998-05-01

The US Department of Energy (DOE) National Center of Excellence for Metals Recycle has recently been established. The vision of this new program is to develop a DOE culture that promotes pollution prevention by considering the recycle and reuse of metal as the first and primary disposition option and burial as a last option. The Center of Excellence takes the approach that unrestricted release of metal is the first priority because it is the most cost-effective disposition pathway. Where this is not appropriate, restricted release, beneficial reuse, and stockpile of ingots are considered. Current recycling activities include the sale of 40,000 tons of scrap metal from the East Tennessee Technology Park (formerly K-25 Plant) K-770 scrap yard, K-1064 surplus equipment and machinery, 7,000 PCB-contaminated drums, 12,000 tons of metal from the Y-l2 scrap yard, and 1,000 metal pallets. In addition, the Center of Excellence is developing a toolbox for project teams that will contain a number of specific tools to facilitate metals recycle. This Internet-based toolbox will include primers, computer programs, and case studies designed to help sites to perform life cycle analysis, perform ALARA (As Low As is Reasonably Achievable) analysis for radiation exposures, provide pollution prevention information and documentation, and produce independent government estimates. The use of these tools is described for two current activities: disposition of scrap metal in the Y-12 scrapyard, and disposition of PCB-contaminated drums

Waste paper recycling opportunities for government action. Vol. 1 summary. Final report

Energy Technology Data Exchange (ETDEWEB)

Bailey, J; Love, P

1978-01-01

This paper presents a summary of analyses of the current and expected waste paper market conditions in Canada with the objective of identifying government initiatives which could permanently increase recovery levels. Emphasis has been placed upon defining short-term, practical measures which take into account the variations in market conditions for each of the major types of secondary fiber: newsprint, fine paper, and corrugated cardboard. Demand, generation and recovery are analyzed for each of these types on a national and regional basis. The opportunities for increased recycling in each region are identified and recommendations are made regarding appropriate government actions. The principal benefits of increased waste paper reclamation and recycling are perceived as: reducing waste management costs, creating new industries, promoting the conserver ethic, conserving forest and energy resources, and reducing pollution. Includes glossary. 1 fig., 5 tabs.

A UK perspective on recycling

International Nuclear Information System (INIS)

Williams, T.

1991-01-01

The United Kingdom, through the recycling of depleted uranium from Magnox reactors into Advanced Gas-cooled Reactor (AGR) fuel, has already recycled significant quantities of reprocessed material in reactors owned by Nuclear Electric plc and Scottish Nuclear Limited. This AGR fuel has been satisfactorily irradiated and discharged over a decade or more, and will be reprocessed in the new Thermal Oxide Reprocessing Plant (THORP), currently under construction in the UK. British Nuclear Fuels plc (BNFL) and the UK Atomic Energy Authority (UKAEA) have also been exploiting the potential of plutonium recycled in mixed oxide (MOX) fuel, which they have been making since 1963. All of the UK nuclear companies are committed to further recycling of Magnox depleted uranium during the 1990s, and it is anticipated that oxide recycling will also become firmly established during the next decade. British Nuclear Fuels and Urenco Ltd, as the providers of fuel cycle services, are developing an infrastructure to close the fuel cycle for oxide nuclear fuel, using both the uranium and plutonium arising from reprocessing. (author)

Recycling Pressure-Sensitive Products

Science.gov (United States)

Jihui Guo; Larry Gwin; Carl Houtman; Mark Kroll; Steven J. Severtson

2012-01-01

The efficient control of contaminants such as metals, plastics, inks and adhesives during the processing of recovered paper products determines the profitability of recycling mills. In fact, it is arguably the most important technical obstacle in expanding the use of recycled paper.1-4 An especially challenging category of contaminants to manage...

DWPF Recycle Evaporator Simulant Tests

International Nuclear Information System (INIS)

Stone, M

2005-01-01

Testing was performed to determine the feasibility and processing characteristics of an evaporation process to reduce the volume of the recycle stream from the Defense Waste Processing Facility (DWPF). The concentrated recycle would be returned to DWPF while the overhead condensate would be transferred to the Effluent Treatment Plant. Various blends of evaporator feed were tested using simulants developed from characterization of actual recycle streams from DWPF and input from DWPF-Engineering. The simulated feed was evaporated in laboratory scale apparatus to target a 30X volume reduction. Condensate and concentrate samples from each run were analyzed and the process characteristics (foaming, scaling, etc) were visually monitored during each run. The following conclusions were made from the testing: Concentration of the ''typical'' recycle stream in DWPF by 30X was feasible. The addition of DWTT recycle streams to the typical recycle stream raises the solids content of the evaporator feed considerably and lowers the amount of concentration that can be achieved. Foaming was noted during all evaporation tests and must be addressed prior to operation of the full-scale evaporator. Tests were conducted that identified Dow Corning 2210 as an antifoam candidate that warrants further evaluation. The condensate has the potential to exceed the ETP WAC for mercury, silicon, and TOC. Controlling the amount of equipment decontamination recycle in the evaporator blend would help meet the TOC limits. The evaporator condensate will be saturated with mercury and elemental mercury will collect in the evaporator condensate collection vessel. No scaling on heating surfaces was noted during the tests, but splatter onto the walls of the evaporation vessels led to a buildup of solids. These solids were difficult to remove with 2M nitric acid. Precipitation of solids was not noted during the testing. Some of the aluminum present in the recycle streams was converted from gibbsite to

A dominant negative mutant of rab5 inhibits infection of cells by foot-and-mouth disease virus; implications for virus entry

DEFF Research Database (Denmark)

Johns, Helen; Berryman, Stephen; Monaghan, Paul

2009-01-01

Foot-and-mouth disease virus (FMDV) can use a number of different integrins (alphavβ1, alphavβ3, alphavβ6, and alphavβ8) as receptors to initiate infection. Infection mediated by alphavβ6 is known to occur by clathrin-mediated endocytosis and is dependent on the acidic pH within endosomes....... On internalization, virus is detected rapidly in early endosomes (EE) and subsequently in perinuclear recycling endosomes (PNRE), but not in late endosomal compartments. Due to the extreme sensitivity of FMDV to acidic pH, it is thought that EE can provide a pH low enough for infection to occur; however, definitive...... proof that infection takes place from within these compartments is still lacking. Here we have investigated the intracellular transport steps required for FMDV infection of IBRS-2 cells, which express vβ8 as their FMDV receptor. These experiments confirmed that FMDV infection mediated by alphavβ8...

Stargazin regulates AMPA receptor trafficking through adaptor protein complexes during long-term depression

Science.gov (United States)

Matsuda, Shinji; Kakegawa, Wataru; Budisantoso, Timotheus; Nomura, Toshihiro; Kohda, Kazuhisa; Yuzaki, Michisuke

2013-11-01

Long-term depression (LTD) underlies learning and memory in various brain regions. Although postsynaptic AMPA receptor trafficking mediates LTD, its underlying molecular mechanisms remain largely unclear. Here we show that stargazin, a transmembrane AMPA receptor regulatory protein, forms a ternary complex with adaptor proteins AP-2 and AP-3A in hippocampal neurons, depending on its phosphorylation state. Inhibiting the stargazin-AP-2 interaction disrupts NMDA-induced AMPA receptor endocytosis, and inhibiting that of stargazin-AP-3A abrogates the late endosomal/lysosomal trafficking of AMPA receptors, thereby upregulating receptor recycling to the cell surface. Similarly, stargazin’s interaction with AP-2 or AP-3A is necessary for low-frequency stimulus-evoked LTD in CA1 hippocampal neurons. Thus, stargazin has a crucial role in NMDA-dependent LTD by regulating two trafficking pathways of AMPA receptors—transport from the cell surface to early endosomes and from early endosomes to late endosomes/lysosomes—through its sequential binding to AP-2 and AP-3A.

Recycling ampersand incineration: Evaluating the choices

International Nuclear Information System (INIS)

Denison, R.A.; Ruston, J.

1993-01-01

Conflicts between proponents of municipal solid waste incineration and advocates of recycling have escalated with efforts to reduce the volume of waste that ends up in landfills. Central to this debate is competition for materials that are both combustible and recyclable. Environmental and economic concerns also play a major role. This book, produced by the Environmental Defense Fund, compares recycling and incineration. It is intended for 'citizens, government officials, and business people who want to help resolve the solid-waste crisis.' The book is divided into three parts: recycling and incineration; health and environmental risk of incineration; and planning, public participation, and environmental review requirements. The book does an excellent job of discussing the benefits of recycling and the pitfalls of incineration. It provides helpful information for identifying questions that should be raised about incineration, but it does not raise similar queries about recycling. There is much worthwhile information here, but the book would be more useful if it identified critical issues for all waste reduction and management options

Enrichment of Phosphatidylethanolamine in Viral Replication Compartments via Co-opting the Endosomal Rab5 Small GTPase by a Positive-Strand RNA Virus.

Directory of Open Access Journals (Sweden)

Kai Xu

2016-10-01

Full Text Available Positive-strand RNA viruses build extensive membranous replication compartments to support replication and protect the virus from antiviral responses by the host. These viruses require host factors and various lipids to form viral replication complexes (VRCs. The VRCs built by Tomato bushy stunt virus (TBSV are enriched with phosphatidylethanolamine (PE through a previously unknown pathway. To unravel the mechanism of PE enrichment within the TBSV replication compartment, in this paper, the authors demonstrate that TBSV co-opts the guanosine triphosphate (GTP-bound active form of the endosomal Rab5 small GTPase via direct interaction with the viral replication protein. Deletion of Rab5 orthologs in a yeast model host or expression of dominant negative mutants of plant Rab5 greatly decreases TBSV replication and prevents the redistribution of PE to the sites of viral replication. We also show that enrichment of PE in the viral replication compartment is assisted by actin filaments. Interestingly, the closely related Carnation Italian ringspot virus, which replicates on the boundary membrane of mitochondria, uses a similar strategy to the peroxisomal TBSV to hijack the Rab5-positive endosomes into the viral replication compartments. Altogether, usurping the GTP-Rab5-positive endosomes allows TBSV to build a PE-enriched viral replication compartment, which is needed to support peak-level replication. Thus, the Rab family of small GTPases includes critical host factors assisting VRC assembly and genesis of the viral replication compartment.

Antimony recycling in the United States in 2000

Science.gov (United States)

Carlin, James F.

2006-01-01

The importance of recycling has become more obvious as concerns about the environment and import dependence have grown in recent years. When materials are recycled, fewer natural resources are consumed, and less waste products go to landfills or pollute the water and air. This study, one of a series of reports on metals recycling in 2000, discusses the flow of antimony from mining through its uses and disposal with emphasis on recycling. In 2000, the recycling efficiency for antimony was estimated to be 89 percent, and the recycling rate was about 20 percent.

Packaging waste recycling in Europe: Is the industry paying for it?

International Nuclear Information System (INIS)

Ferreira da Cruz, Nuno; Ferreira, Sandra; Cabral, Marta; Simões, Pedro; Marques, Rui Cunha

2014-01-01

Highlights: • We study the recycling schemes of France, Germany, Portugal, Romania and the UK. • The costs and benefits of recycling are compared for France, Portugal and Romania. • The balance of costs and benefits depend on the perspective (strictly financial/economic). • Financial supports to local authorities ought to promote cost-efficiency. - Abstract: This paper describes and examines the schemes established in five EU countries for the recycling of packaging waste. The changes in packaging waste management were mainly implemented since the Directive 94/62/EC on packaging and packaging waste entered into force. The analysis of the five systems allowed the authors to identify very different approaches to cope with the same problem: meet the recovery and recycling targets imposed by EU law. Packaging waste is a responsibility of the industry. However, local governments are generally in charge of waste management, particularly in countries with Green Dot schemes or similar extended producer responsibility systems. This leads to the need of establishing a system of financial transfers between the industry and the local governments (particularly regarding the extra costs involved with selective collection and sorting). Using the same methodological approach, the authors also compare the costs and benefits of recycling from the perspective of local public authorities for France, Portugal and Romania. Since the purpose of the current paper is to take note of who is paying for the incremental costs of recycling and whether the industry (i.e. the consumer) is paying for the net financial costs of packaging waste management, environmental impacts are not included in the analysis. The work carried out in this paper highlights some aspects that are prone to be improved and raises several questions that will require further research. In the three countries analyzed more closely in this paper the industry is not paying the net financial cost of packaging waste

Packaging waste recycling in Europe: Is the industry paying for it?

Energy Technology Data Exchange (ETDEWEB)

Ferreira da Cruz, Nuno, E-mail: nunocruz@ist.utl.pt; Ferreira, Sandra; Cabral, Marta; Simões, Pedro; Marques, Rui Cunha

2014-02-15

Highlights: • We study the recycling schemes of France, Germany, Portugal, Romania and the UK. • The costs and benefits of recycling are compared for France, Portugal and Romania. • The balance of costs and benefits depend on the perspective (strictly financial/economic). • Financial supports to local authorities ought to promote cost-efficiency. - Abstract: This paper describes and examines the schemes established in five EU countries for the recycling of packaging waste. The changes in packaging waste management were mainly implemented since the Directive 94/62/EC on packaging and packaging waste entered into force. The analysis of the five systems allowed the authors to identify very different approaches to cope with the same problem: meet the recovery and recycling targets imposed by EU law. Packaging waste is a responsibility of the industry. However, local governments are generally in charge of waste management, particularly in countries with Green Dot schemes or similar extended producer responsibility systems. This leads to the need of establishing a system of financial transfers between the industry and the local governments (particularly regarding the extra costs involved with selective collection and sorting). Using the same methodological approach, the authors also compare the costs and benefits of recycling from the perspective of local public authorities for France, Portugal and Romania. Since the purpose of the current paper is to take note of who is paying for the incremental costs of recycling and whether the industry (i.e. the consumer) is paying for the net financial costs of packaging waste management, environmental impacts are not included in the analysis. The work carried out in this paper highlights some aspects that are prone to be improved and raises several questions that will require further research. In the three countries analyzed more closely in this paper the industry is not paying the net financial cost of packaging waste

Design study on advanced nuclear fuel recycle system. Conceptual design study of recycle system using molten salt

International Nuclear Information System (INIS)

Kasai, Y.; Kakehi, I.; Moro, T.; Higashi, T.; Tobe, K.; Kawamura, F.; Yonezawa, S.; Yoshiuji, T.

1998-10-01

Advanced recycle system engineering group of OEC (Oarai Engineering Center) has being carried out a design study of the advanced nuclear fuel recycle system using molten salt (electro-metallurgical process). This system is aiming for improvements of fuel cycle economy and reduction of environmental burden (MA recycles, Minimum of radioactive waste disposal), and also improvement of safety and nuclear non-proliferation. This report describes results of the design study that has been continued since December 1996. (1) A design concept of the advanced nuclear fuel recycle system, that is a module type recycles system of pyrochemical reprocessing and fuel re-fabrication was studied. The module system has advantage in balance of Pu recycle where modules are constructed in coincidence with the construction plan of nuclear power plants, and also has flexibility for technology progress. A demonstration system, minimum size of the above module, was studies. This system has capacity of 10 tHM/y and is able to demonstrate recycle technology of MOX fuel, metal fuel and nitride fuel. (2) Each process of the system, which are pyrochemical electrorefining system, cathode processor, de-cladding system, waste disposal system, etc., were studied. In this study, capacity of an electrorefiner was discussed, and vitrification experiment of molten salt using lead-boric acid glass was conducted. (3) A hot cell system and material handling system of the demonstration system was studied. A robot driven by linear motor was studied for the handling system, and an arrangement plan of the cell system was made. Criticality analysis in the cell system and investigation of material accountancy system of the recycle plant were also made. This design study will be continued in coincidence with design study of reactor and fuel, aiming to establish the concept of FBR recycle system. (author)

Mechanical and chemical recycling of solid plastic waste.

Science.gov (United States)

Ragaert, Kim; Delva, Laurens; Van Geem, Kevin

2017-11-01

This review presents a comprehensive description of the current pathways for recycling of polymers, via both mechanical and chemical recycling. The principles of these recycling pathways are framed against current-day industrial reality, by discussing predominant industrial technologies, design strategies and recycling examples of specific waste streams. Starting with an overview on types of solid plastic waste (SPW) and their origins, the manuscript continues with a discussion on the different valorisation options for SPW. The section on mechanical recycling contains an overview of current sorting technologies, specific challenges for mechanical recycling such as thermo-mechanical or lifetime degradation and the immiscibility of polymer blends. It also includes some industrial examples such as polyethylene terephthalate (PET) recycling, and SPW from post-consumer packaging, end-of-life vehicles or electr(on)ic devices. A separate section is dedicated to the relationship between design and recycling, emphasizing the role of concepts such as Design from Recycling. The section on chemical recycling collects a state-of-the-art on techniques such as chemolysis, pyrolysis, fluid catalytic cracking, hydrogen techniques and gasification. Additionally, this review discusses the main challenges (and some potential remedies) to these recycling strategies and ground them in the relevant polymer science, thus providing an academic angle as well as an applied one. Copyright © 2017 Elsevier Ltd. All rights reserved.

Filter Backwash Recycling Rule Documents

Science.gov (United States)

The purpose of the FBRR is to require (PWSs) to review their recycle practices and, where appropriate, work with the state Primacy Agency to make any necessary changes to recycle practices that may compromise microbial control.

Research on the recycling industry development model for typical exterior plastic components of end-of-life passenger vehicle based on the SWOT method.

Science.gov (United States)

Zhang, Hongshen; Chen, Ming

2013-11-01

In-depth studies on the recycling of typical automotive exterior plastic parts are significant and beneficial for environmental protection, energy conservation, and sustainable development of China. In the current study, several methods were used to analyze the recycling industry model for typical exterior parts of passenger vehicles in China. The strengths, weaknesses, opportunities, and challenges of the current recycling industry for typical exterior parts of passenger vehicles were analyzed comprehensively based on the SWOT method. The internal factor evaluation matrix and external factor evaluation matrix were used to evaluate the internal and external factors of the recycling industry. The recycling industry was found to respond well to all the factors and it was found to face good developing opportunities. Then, the cross-link strategies analysis for the typical exterior parts of the passenger car industry of China was conducted based on the SWOT analysis strategies and established SWOT matrix. Finally, based on the aforementioned research, the recycling industry model led by automobile manufacturers was promoted. Copyright © 2013 Elsevier Ltd. All rights reserved.

Coal liquefaction with preasphaltene recycle

Science.gov (United States)

Weimer, Robert F.; Miller, Robert N.

1986-01-01

A coal liquefaction system is disclosed with a novel preasphaltene recycle from a supercritical extraction unit to the slurry mix tank wherein the recycle stream contains at least 90% preasphaltenes (benzene insoluble, pyridine soluble organics) with other residual materials such as unconverted coal and ash. This subject process results in the production of asphaltene materials which can be subjected to hydrotreating to acquire a substitute for No. 6 fuel oil. The preasphaltene-predominant recycle reduces the hydrogen consumption for a process where asphaltene material is being sought.

U.S. Department of Energy National Center of Excellence for Metals Recycle

International Nuclear Information System (INIS)

Adams, V.; Bennett, M.; Bishop, L.

1998-06-01

The US Department of Energy (DOE) National Center of Excellence for Metals Recycle has recently been established. The vision of this new program is to develop a DOE culture that promotes pollution prevention by considering the recycle and reuse of metal as the first and primary disposition option and burial as a last option. The Center of Excellence takes the approach that unrestricted release of metal is the first priority because it is the most cost-effective disposition pathway. Where this is not appropriate, restricted release, beneficial reuse, and stockpile of ingots are considered. The Center has gotten off to a fast start. Current recycling activities include the sale of 40,000 tons of scrap metal from the East Tennessee Technology Park (formerly K-25 Plant) K-770 scrap yard, K-1064 surplus equipment and machinery, 7,000 PCB-contaminated drums, 12,000 tons of metal from the Y-12 scrap yard, and 1,000 metal pallets. In addition, the Center of Excellence is developing a toolbox for project teams that will contain a number of specific tools to facilitate metals recycle. This Internet-based toolbox will include primers, computer software, and case studies designed to help sites to perform life cycle analysis, perform ALARA (As Low As is Reasonably Achievable) analysis for radiation exposures, produce pollution prevention information and documentation, manage their materials inventory, produce independent government estimates, and implement sale/service contracts. The use of these tools is described for two current activities: disposition of scrap metal in the Y-12 scrap yard, and disposition of PCB-contaminated drums. Members of the Center look forward to working with all DOE sites, regulatory authorities, the private sector, and other stakeholders to achieve the metals recycle goals

Overview of HTGR fuel recycle

International Nuclear Information System (INIS)

Notz, K.J.

1976-01-01

An overview of HTGR fuel recycle is presented, with emphasis placed on reprocessing and fuel kernel refabrication. Overall recycle operations include (1) shipment and storage, (2) reprocessing, (3) refabrication, (4) waste handling, and (5) accountability and safeguards

Sustaining the environment through recycling: an empirical study.

Science.gov (United States)

Ramayah, T; Lee, Jason Wai Chow; Lim, Shuwen

2012-07-15

This paper examines the determinants of recycling behaviour among 200 university students from the perspective of the theory of planned behaviour (TPB). Data was analysed using Structural Equation Modelling technique. Findings indicate that environmental awareness was significantly related to attitude towards recycling, whilst attitude and social norms had significant impact on recycling behaviour. However, convenience and cost of recycling were not significant reasons for recycling. The study has enhanced the understanding of the determinants of recycling behaviour and has implications for schools and governmental agencies in educating and encouraging positive recycling behaviour. It also confirms the appropriateness of the TPB in examining studies of this nature. Further suggestions for future research are offered. Copyright © 2012 Elsevier Ltd. All rights reserved.

Recycling fusion materials

International Nuclear Information System (INIS)

Ooms, L.

2005-01-01

The inherent safety and environmental advantages of fusion power in comparison with other energy sources play an important role in the public acceptance. No waste burden for future generations is therefore one of the main arguments to decide for fusion power. The waste issue has thus been studied in several documents and the final conclusion of which it is stated that there is no permanent disposal waste needed if recycling is applied. But recycling of fusion reactor materials is far to be obvious regarding mostly the very high specific activity of the materials to be handled, the types of materials and the presence of tritium. The main objective of research performed by SCK-CEN is to study the possible ways of recycling fusion materials and analyse the challenges of the materials management from fusion reactors, based on current practices used in fission reactors and the requirements for the manufacture of fusion equipment

Extracellular delivery induced by ultrasound and microbubbles in cells

Science.gov (United States)

Hussein, Farah; Antonescu, Costin; Karshafian, Raffi

2017-03-01

Ultrasound and microbubble treatment (USMB) can enhance the intracellular uptake of molecules, which otherwise would be excluded from the cell, through USMB-mediated transient membrane disruption and through enhanced endocytosis. However, the effect of USMB on the outward movement of molecules from cells is not well understood. This study investigates the effects of USMB on the release of molecules from various cellular compartments including cytoplasm, lysosomes, and recycling endosomes. In vitro ARPE-19 (RPE henceforth) cells were loaded with Alexa fluor-labeled transferrin as a marker for recycling endosomes, LAMP-1 antibody was used to detect the fusion of lysosomes with the plasma membrane, GFP-transfected RPE cells were used to examine the release of GFP from the cytoplasm, and 7-AAD was used to assess cell viability. Subsequently, cells were exposed to USMB (106 cells/mL, 300 kPa peak negative pressure, 1 min treatment duration, and 20 µL/mL Definity microbubbles). Following USMB, the release of the fluorescent markers was examined at 1.5, 11.5, and 21.5 minutes from the start of USMB. The mean fluorescent intensity (MFI) of untreated and USMB treated samples were measured using flow cytometry. USMB increased the extracellular delivery of GFP molecules from the cytoplasm; the MFI in USMB treated GFP-transfected RPE cells decreased by 17% in viable cells and this MFI decreased by 70% in non-viable cells. This could be due to diffusion of GFP through the membrane disruptions induced by USMB. Additionally, the MFI of viable cells stained with LAMP-1 antibody increased by 50% and this increase was 15 folds in the non-viable cells indicating lysosome exocytosis as a mechanism for membrane repair. Furthermore, the MFI of cells loaded with fluorescent transferrin decreased by 22% after USMB treatment in viable cells, indicating a significant increase in transferrin recycling to the cell membrane. However, the increased recycling was not statistically significant

Recycling of polymers: a review.

Science.gov (United States)

Ignatyev, Igor A; Thielemans, Wim; Vander Beke, Bob

2014-06-01

Plastics are inexpensive, easy to mold, and lightweight. These and many other advantages make them very promising candidates for commercial applications. In many areas, they have substantially suppressed traditional materials. However, the problem of recycling still is a major challenge. There are both technological and economic issues that restrain the progress in this field. Herein, a state-of-art overview of recycling is provided together with an outlook for the future by using popular polymers such as polyolefins, poly(vinyl chloride), polyurethane, and poly(ethylene terephthalate) as examples. Different types of recycling, primary, secondary, tertiary, quaternary, and biological recycling, are discussed together with related issues, such as compatibilization and cross-linking. There are various projects in the European Union on research and application of these recycling approaches; selected examples are provided in this article. Their progress is mirrored by granted patents, most of which have a very limited scope and narrowly cover certain technologies. Global introduction of waste utilization techniques to the polymer market is currently not fully developed, but has an enormous potential. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Long-term aging of recycled binders : [summary].

Science.gov (United States)

2015-10-01

At 80 million tons a year representing more than 80% of all milled asphalt pavement : asphalt paving is Americas most recycled material. Asphalt can be recycled in place, which is : very cost effective; however, aging of recycled binder ca...

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

Science.gov (United States)

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

2009-05-01

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

Waste management considerations in HTGR recycle operations

International Nuclear Information System (INIS)

Pence, D.T.; Shefcik, J.J.; Heath, C.A.

1975-01-01

Waste management considerations in the recycle of HTGR fuel are different from those encountered in the recycle of LWR fuel. The types of waste associated with HTGR recycle operations are discussed, and treatment methods for some of the wastes are described

Concrete Waste Recycling Process for High Quality Aggregate

International Nuclear Information System (INIS)

Ishikura, Takeshi; Fujii, Shin-ichi

2008-01-01

Large amount of concrete waste generates during nuclear power plant (NPP) dismantling. Non-contaminated concrete waste is assumed to be disposed in a landfill site, but that will not be the solution especially in the future, because of decreasing tendency of the site availability and natural resources. Concerning concrete recycling, demand for roadbeds and backfill tends to be less than the amount of dismantled concrete generated in a single rural site, and conventional recycled aggregate is limited of its use to non-structural concrete, because of its inferior quality to ordinary natural aggregate. Therefore, it is vital to develop high quality recycled aggregate for general uses of dismantled concrete. If recycled aggregate is available for high structural concrete, the dismantling concrete is recyclable as aggregate for industry including nuclear field. Authors developed techniques on high quality aggregate reclamation for large amount of concrete generated during NPP decommissioning. Concrete of NPP buildings has good features for recycling aggregate; large quantity of high quality aggregate from same origin, record keeping of the aggregate origin, and little impurities in dismantled concrete such as wood and plastics. The target of recycled aggregate in this development is to meet the quality criteria for NPP concrete as prescribed in JASS 5N 'Specification for Nuclear Power Facility Reinforced Concrete' and JASS 5 'Specification for Reinforced Concrete Work'. The target of recycled aggregate concrete is to be comparable performance with ordinary aggregate concrete. The high quality recycled aggregate production techniques are assumed to apply for recycling for large amount of non-contaminated concrete. These techniques can also be applied for slightly contaminated concrete dismantled from radiological control area (RCA), together with free release survey. In conclusion: a technology on dismantled concrete recycling for high quality aggregate was developed

What do we know about metal recycling rates?

Science.gov (United States)

Graedel, T.E.; Allwood, J.; Birat, J.-P.; Buchert, M.; Hageluken, C.; Reck, B.K.; Sibley, S.F.; Sonnemann, G.

2011-01-01

The recycling of metals is widely viewed as a fruitful sustainability strategy, but little information is available on the degree to which recycling is actually taking place. This article provides an overview on the current knowledge of recycling rates for 60 metals. We propose various recycling metrics, discuss relevant aspects of recycling processes, and present current estimates on global end-of-life recycling rates (EOL-RR; i.e., the percentage of a metal in discards that is actually recycled), recycled content (RC), and old scrap ratios (OSRs; i.e., the share of old scrap in the total scrap flow). Because of increases in metal use over time and long metal in-use lifetimes, many RC values are low and will remain so for the foreseeable future. Because of relatively low efficiencies in the collection and processing of most discarded products, inherent limitations in recycling processes, and the fact that primary material is often relatively abundant and low-cost (which thereby keeps down the price of scrap), many EOL-RRs are very low: Only for 18 metals (silver, aluminum, gold, cobalt, chromium, copper, iron, manganese, niobium, nickel, lead, palladium, platinum, rhenium, rhodium, tin, titanium, and zinc) is the EOL-RR above 50% at present. Only for niobium, lead, and ruthenium is the RC above 50%, although 16 metals are in the 25% to 50% range. Thirteen metals have an OSR greater than 50%. These estimates may be used in considerations of whether recycling efficiencies can be improved; which metric could best encourage improved effectiveness in recycling; and an improved understanding of the dependence of recycling on economics, technology, and other factors. ?? 2011 by Yale University.

Energetic conditions of effective recycling of composite castings

OpenAIRE

J. Jackowski

2009-01-01

The most reasonable way of recycling the metal composite materials consists in separation of the components. In case of the composites with saturated reinforcement it is the only recycling method. The process of separation of the components always undergoes in the presence of an additional liquid phase called a recycling medium. In a three-phase system including the material of composite reinforcement – liquid composite matrix – liquid recycling medium, an important role for the recycling pro...

Is Municipal Solid Waste Recycling Economically Efficient?

Science.gov (United States)

Lavee, Doron

2007-12-01

It has traditionally been argued that recycling municipal solid waste (MSW) is usually not economically viable and that only when externalities, long-term dynamic considerations, and/or the entire product life cycle are taken into account, recycling becomes worthwhile from a social point of view. This article explores the results of a wide study conducted in Israel in the years 2000 2004. Our results reveal that recycling is optimal more often than usually claimed, even when externality considerations are ignored. The study is unique in the tools it uses to explore the efficiency of recycling: a computer-based simulation applied to an extensive database. We developed a simulation for assessing the costs of handling and treating MSW under different waste-management systems and used this simulation to explore possible cost reductions obtained by designating some of the waste (otherwise sent to landfill) to recycling. We ran the simulation on data from 79 municipalities in Israel that produce over 60% of MSW in Israel. For each municipality, we were able to arrive at an optimal method of waste management and compare the costs associated with 100% landfilling to the costs born by the municipality when some of the waste is recycled. Our results indicate that for 51% of the municipalities, it would be efficient to adopt recycling, even without accounting for externality costs. We found that by adopting recycling, municipalities would be able to reduce direct costs by an average of 11%. Through interviews conducted with representatives of municipalities, we were also able to identify obstacles to the utilization of recycling, answering in part the question of why actual recycling levels in Israel are lower than our model predicts they should be.

India's ship recycling trade-off

NARCIS (Netherlands)

Worrell, E.; Athanasopoulou, V.

2014-01-01

The special nature of India's steel industry lends particular importance to ship recycling as a source of scrap. Ship recycling in upgraded 'green' facilities can substitute other 'dirty' ironmaking processes, resulting in energy savings and air pollutant emission reductions for the Indian steel

Mox fuels recycling

International Nuclear Information System (INIS)

Gay, A.

1998-01-01

This paper will firstly emphasis that the first recycling of plutonium is already an industrial reality in France thanks to the high degree of performance of La Hague and MELOX COGEMA's plants. Secondly, recycling of spent Mixed OXide fuel, as a complete MOX fuel cycle, will be demonstrated through the ability of the existing plants and services which have been designed to proceed with such fuels. Each step of the MOX fuel cycle concept will be presented: transportation, reception and storage at La Hague and steps of spent MOX fuel reprocessing. (author)

Recycling of Reinforced Plastics

Science.gov (United States)

Adams, R. D.; Collins, Andrew; Cooper, Duncan; Wingfield-Digby, Mark; Watts-Farmer, Archibald; Laurence, Anna; Patel, Kayur; Stevens, Mark; Watkins, Rhodri

2014-02-01

This work has shown is that it is possible to recycle continuous and short fibre reinforced thermosetting resins while keeping almost the whole of the original material, both fibres and matrix, within the recyclate. By splitting, crushing hot or cold, and hot forming, it is possible to create a recyclable material, which we designate a Remat, which can then be used to remanufacture other shapes, examples of plates and tubes being demonstrated. Not only can remanufacturing be done, but it has been shown that over 50 % of the original mechanical properties, such as the E modulus, tensile strength, and interlaminar shear strength, can be retained. Four different forms of composite were investigated, a random mat Glass Fibre Reinforced Plastic (GFRP) bathroom component and boat hull, woven glass and carbon fibre cloth impregnated with an epoxy resin, and unidirectional carbon fibre pre-preg. One of the main factors found to affect composite recyclability was the type of resin matrix used in the composite. Thermoset resins tested were shown to have a temperature range around the Glass Transition Temperature (Tg) where they exhibit ductile behaviour, hence aiding reforming of the material. The high-grade carbon fibre prepreg was found to be less easy to recycle than the woven of random fibre laminates. One method of remanufacturing was by heating the Remat to above its glass transition temperature, bending it to shape, and then cooling it. However, unless precautions are taken, the geometric form may revert. This does not happen with the crushed material.

A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

Directory of Open Access Journals (Sweden)

Gordon Ng

Full Text Available BACKGROUND: Nitrogen (N(2 fixation also yields hydrogen (H(2 at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2 as sole N-source bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase, has nevertheless been presumed responsible for recycling such endogenous hydrogen. METHODS AND FINDINGS: As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase. An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. CONCLUSIONS: Representative of aerobic N(2-fixing and H(2-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2 respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2, specifically that produced by N(2 fixation. To benefit human civilization, H(2 has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As

A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

Science.gov (United States)

Ng, Gordon; Tom, Curtis G S; Park, Angela S; Zenad, Lounis; Ludwig, Robert A

2009-01-01

Nitrogen (N(2)) fixation also yields hydrogen (H(2)) at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2) as sole N-source) bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase), has nevertheless been presumed responsible for recycling such endogenous hydrogen. As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase) was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase). An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2)-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. Representative of aerobic N(2)-fixing and H(2)-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2) respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2), specifically that produced by N(2) fixation. To benefit human civilization, H(2) has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As such, the reversible, group-4 Ni,Fe-hydrogenases, such

76 FR 71861 - America Recycles Day, 2011

Science.gov (United States)

2011-11-18

... electronics annually, and without following proper recycling and management practices, the disposal of our old..., and prevent the recovery and reuse of valuable resources. For the well- being of our people and our..., management, and recycling that will accelerate our burgeoning electronics recycling market and create jobs...

On the logistics of recycling : an introduction

NARCIS (Netherlands)

Flapper, S.D.P.

1993-01-01

An overview is given of the different logistic aspects of recycling, where recycling denotes "All the activities required for the reuse of materials and (semi-)finished products after they are no longer used by their last user." Special attention is paid to the forced recycling of durable

Hydrogen recycle modeling in transport codes

International Nuclear Information System (INIS)

Howe, H.C.

1979-01-01

The hydrogen recycling models now used in Tokamak transport codes are reviewed and the method by which realistic recycling models are being added is discussed. Present models use arbitrary recycle coefficients and therefore do not model the actual recycling processes at the wall. A model for the hydrogen concentration in the wall serves two purposes: (1) it allows a better understanding of the density behavior in present gas puff, pellet, and neutral beam heating experiments; and (2) it allows one to extrapolate to long pulse devices such as EBT, ISX-C and reactors where the walls are observed or expected to saturate. Several wall models are presently being studied for inclusion in transport codes

Exosomes derived from human macrophages suppress endothelial cell migration by controlling integrin trafficking.

Science.gov (United States)

Lee, Hee Doo; Kim, Yeon Hyang; Kim, Doo-Sik

2014-04-01

Integrin trafficking, including internalization, recycling, and lysosomal degradation, is crucial for the regulation of cellular functions. Exosomes, nano-sized extracellular vesicles, are believed to play important roles in intercellular communications. This study demonstrates that exosomes released from human macrophages negatively regulate endothelial cell migration through control of integrin trafficking. Macrophage-derived exosomes promote internalization of integrin β1 in primary HUVECs. The internalized integrin β1 persistently accumulates in the perinuclear region and is not recycled back to the plasma membrane. Experimental results indicate that macrophage-derived exosomes stimulate trafficking of internalized integrin β1 to lysosomal compartments with a corresponding decrease in the integrin destined for recycling endosomes, resulting in proteolytic degradation of the integrin. Moreover, ubiquitination of HUVEC integrin β1 is enhanced by the exosomes, and exosome-mediated integrin degradation is blocked by bafilomycin A, a lysosomal degradation inhibitor. Macrophage-derived exosomes were also shown to effectively suppress collagen-induced activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase signaling pathway and HUVEC migration, which are both dependent on integrin β1. These observations provide new insight into the functional significance of exosomes in the regulation of integrin trafficking. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The position of the Bureau of International Recycling on unwanted radioactivity and scrap

International Nuclear Information System (INIS)

Neenan, P.; Bartley, R.

1999-01-01

The Bureau of International Recycling (BIR) speaks as the unified voice of the international recycling industry on a world level, promoting free-trade in non-hazardous recyclables and the increased usage of recycled goods. In October 1998, BIR presented its 'Guide to Radioactivity' to its members in order to explain the basics of radioactivity, to help determine what actions material recyclers should take and to provide an initial contact guide for summoning expert help. The Guide was promulgated together with a survey questionnaire; initial results of this survey have been collated. BIR members do not trade in radioactive materials and do not wish to receive hazardous radioactive contaminated materials. Furthermore it is very uncommon to find this unwelcome and unwanted material. The burden of handling contaminated materials should not rest on the recycling industry alone, however, as the hazards associated with processing contaminated materials are potentially extreme, recyclers have a role to help safeguard both their workforce and subsequent customers. Some BIR members have invested in in-plant detection equipment of various types from a variety of suppliers to detect contamination from 'lost' sources or from naturally occurring radioactive materials (NORM). Strict State control on sources in use would be very welcome to limit these ' losses'. Not all BIR members are in a position to provide or operate detection equipment; therefore, BIR would welcome States providing detection and control at sea ports and at border checkpoints on railways and roads. Some states have proposed permitting requirements for the handling of radioactive contaminated materials and have tried, inappropriately, to impose on the mainstream recycling industry, permits designed for the specialist decontamination or decommissioning industry. Current discussions on applying the transport of dangerous goods regulations would lead to unnecessary and impracticable additional restraints to

Exposure to hazardous substances in Cathode Ray Tube (CRT) recycling sites in France

Energy Technology Data Exchange (ETDEWEB)

Lecler, Marie-Thérèse, E-mail: marie-therese.lecler@inrs.fr; Zimmermann, François; Silvente, Eric; Clerc, Frédéric; Chollot, Alain; Grosjean, Jérôme

2015-05-15

Highlights: • Chemical risks were assessed in the nine cathode ray tube screens recycling facilities. • The main hazardous agents are dust containing lead, cadmium, barium and yttrium. • Exposure and pollutant levels are described for different operations and processes. • All the operations and processes are concerned by significant levels of pollutants. • We suggest recommendations to reduce chemical risk. - Abstract: The Waste Electrical and Electronic Equipment (WEEE) or e-waste recycling sector has grown considerably in the last fifteen years due to the ever shorter life cycles of consumables and an increasingly restrictive policy context. Cathode Ray Tubes (CRTs) from used television and computer screens represent one of the main sources of e-waste. CRTs contain toxic materials such as lead, cadmium, barium, and fluorescent powders which can be released if recycling of CRTs is not appropriate. Exposure to these harmful substances was assessed in nine workshops where CRT screens are treated. Particulate exposure levels were measured using a gravimetric method and metals were analysed by plasma emission spectrometry. The maximum levels of worker exposure were 8.8 mg/m{sup 3}, 1504.3 μg/m{sup 3}, 434.9 μg/m{sup 3}, 576.3 μg/m{sup 3} and 2894.3 μg/m{sup 3} respectively for inhalable dust, barium, cadmium, lead and yttrium. The maximum levels of airborne pollutants in static samples were 39.0 mg/m{sup 3}, 848.2 μg/m{sup 3}, 698.4 μg/m{sup 3}, 549.3 μg/m{sup 3} and 3437.9 μg/m{sup 3} for inhalable dust, barium, cadmium, lead and yttrium. The most harmful operations were identified, and preventive measures for reducing the chemical risk associated with screen recycling were proposed. Workplace measurements were used to define recommendations for reducing the chemical risks in CRT screens recycling facilities and for promoting the design and development of “clean and safe” processes in emerging recycling channels.

Recycling of nonferrous metals from waste materials

Energy Technology Data Exchange (ETDEWEB)

Urban, A

1982-02-01

Recycling of metals was one of the 9 central subjects of the international symposium on 'Materials and Energy from Refuse', held in Antwerpen on October 20 to 22, 1981. Six of 65 poster sessions papers were on metal recycling; four of them discussed the recycling of nonferrous metals.

Recycling behaviour in healthcare: waste handling at work.

Science.gov (United States)

Vogt, Joachim; Nunes, Katia R A

2014-01-01

This article reviews the motivational factors for environmental behaviour in general, presenting a case study on recycling disposable plastics in hospitals. Results show that 90% of over 600 employees from six analysed hospitals in Germany reported that the recycling of disposable plastics on the wards makes sense from an environmental and economic point of view. The case study reports an assessment of recycling attitudes and problems of hospital staff, mainly nurses. Employees in eco-certified hospitals were much more satisfied and reported fewer problems with the recycling system. The gender effect was significant only for saving energy, while age correlated with nearly all reported pro-environmental behaviour at home. At work, the mere introduction of a recycling system was insufficient to achieve good recycling results. Based on the study findings, recommendations are given aimed at improving the safety and sustainability of the recycling system.

Space Plastic Recycling System, Phase I

Data.gov (United States)

National Aeronautics and Space Administration — Techshot's proposed Space Plastic Recycler (SPR) is an automated closed loop plastic recycling system that allows the automated conversion of disposable ISS...

Probabilistic Analysis of Structural Member from Recycled Aggregate Concrete

Science.gov (United States)

Broukalová, I.; Šeps, K.

2017-09-01

The paper aims at the topic of sustainable building concerning recycling of waste rubble concrete from demolition. Considering demands of maximising recycled aggregate use and minimising of cement consumption, composite from recycled concrete aggregate was proposed. The objective of the presented investigations was to verify feasibility of the recycled aggregate cement based fibre reinforced composite in a structural member. Reliability of wall from recycled aggregate fibre reinforced composite was assessed in a probabilistic analysis of a load-bearing capacity of the wall. The applicability of recycled aggregate fibre reinforced concrete in structural applications was demonstrated. The outcomes refer to issue of high scatter of material parameters of recycled aggregate concretes.

Bottle & Can Recycling in Denmark: addressing issues and optimizing the recycling rate

OpenAIRE

Carnevale, Alessandro; Larsen, Lucas; Tetens, Simon W.

2015-01-01

The research is conducted in the landscape of environmental care and consumer behaviour. This paper explores the different dynamics revolving around the recycling of plastic bottles and aluminium cans in Denmark. The aim is to shed light on the possible impediments people might encounter when attempting to hand back their deposit marked bottle to the allocated facilities as well as illuminating what the main forces involved in encouraging or inhibiting people from recycling are. The ultimate ...

Economics and recycling

Energy Technology Data Exchange (ETDEWEB)

Butlin, J A

1977-06-01

The current state of recycling technology could appear to be a question of supply and demand, first for storage, disposal, and reclamation facilities, and secondly, for reclaimed materials. If supply and demand are to be relied upon as an environmental policy tool, several conditions need to exist within the economy: supply data for storage and disposal facilities should reflect the full social cost of their use for this purpose relative to any other; demand data for the use of storage facilities must reflect the full social benefit of having waste go through one channel rather than some other; demand for and supply of reclaimed materials for recycling must reflect the full costs and benefits of rechanneling them back into production or consumption; and the markets for products competitive to recycled raw materials (mainly virgin raw materials) should reflect full social costs and benefits, as should the markets for the alternative uses of storage and disposal facilities. If these conditions are met (in addition to a few technical ones), then the problem of waste management will not arise. (MCW)