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Sample records for metalloprotease invadolysin localizes

  1. Intracellular co-localization of trypsin-2 and matrix metalloprotease-9: possible proteolytic cascade of trypsin-2, MMP-9 and enterokinase in carcinoma.

    PubMed

    Vilen, Suvi-Tuuli; Nyberg, Pia; Hukkanen, Mika; Sutinen, Meeri; Ylipalosaari, Merja; Bjartell, Anders; Paju, Annukka; Haaparanta, Virpi; Stenman, Ulf-Håkan; Sorsa, Timo; Salo, Tuula

    2008-02-15

    Tumor-associated trypsin-2 and matrix metalloprotease-9 (MMP-9) are associated with cancer, particularly with invasive squamous cell carcinomas. They require activation for catalytical competence via proteolytic cascades. One cascade is formed by enterokinase, trypsin-2 and MMP-9; enterokinase activates trypsinogen-2 to trypsin-2, which is an efficient proMMP-9 activator. We describe here that oral squamous cell carcinomas express all members of this cascade: MMP-9, trypsin-2 and enterokinase. The expression of enterokinase in a carcinoma cell line not derived from the duodenum was shown here for the first time. Enterokinase directly cleaved proMMP-9 at the Lys65-Ser66 site, but failed to activate it in vitro. We demonstrated by confocal microscopy that MMP-9 and trypsin-2 co-localized in intracellular vesicles of the carcinoma cells. This co-localization of trypsin-2 and MMP-9 resulted in intracellular proMMP-9 processing that represented fully or partially activated MMP-9. However, although both proteases were present also in various bone tumor tissues, MMP-9 and trypsin-2 never co-localized at the cellular level in these tissues. This suggests that the intracellular vesicular co-localization, storage and possible activation of these proteases may be a unique feature for aggressive epithelial tumors, such as squamous cell carcinomas, but not for tumors of mesenchymal origin.

  2. Metalloproteases and tendinopathy.

    PubMed

    Del Buono, Angelo; Oliva, Francesco; Osti, Leonardo; Maffulli, Nicola

    2013-01-01

    Matrix metalloproteinases (MMP) are involved in the development of tendinopathy. These potent enzymes completely degrade all components of the connective tissue, modify the extracellular matrix (ECM), and mediate the development of painful tendinopathy. To control the local activity of activated proteinases, the same cells produce tissue inhibitors of metalloproteinases (TIMP). These latter bind to the enzyme and prevent degradation. The balance between the activities of MMPs and TIMPs regulates tendon remodeling, whereas an imbalance produces a collagen dis-regulation and disturbances in tendons. ADAMs (a disintegrin and metalloproteinase) are cell membrane-linked enzymes with proteolytic and cell signaling functions. ADAMTSs (ADAM with thrombospondin motifs) are secreted into the circulation and constitute a heterogenous family of proteases with both anabolic and catabolic functions. Further studies are needed to better define the mechanism of action, and whether these new strategies are safe and effective in larger models.

  3. Membrane type-1 matrix metalloprotease-independent activation of pro-matrix metalloprotease-2 by proprotein convertases.

    PubMed

    Koo, Bon-Hun; Kim, Hee-Hyun; Park, Michael Y; Jeon, Ok-Hee; Kim, Doo-Sik

    2009-11-01

    Matrix metalloprotease-2 is implicated in many biological processes and degrades extracellular and non-extracellular matrix molecules. Matrix metalloprotease-2 maintains a latent state through a cysteine-zinc ion pairing which, when disrupted, results in full enzyme activation. This pairing can be disrupted by a conformational change or cleavage within the propeptide. The best known activation mechanism for pro-matrix metalloprotease-2 occurs via cleavage of the propeptide by membrane type-1 matrix metalloprotease. However, significant residual activation of pro-matrix metalloprotease-2 is seen in membrane type-1 matrix metalloprotease knockout mice and in fibroblasts treated with metalloprotease inhibitors. These findings indicate the presence of a membrane type-1 matrix metalloprotease-independent activation mechanism for pro-matrix metalloprotease-2 in vivo, which prompted us to explore an alternative activation mechanism for pro-matrix metalloprotese-2. In this study, we demonstrate membrane type-1 matrix metalloprotease-independent propeptide processing of matrix metalloprotease-2 in HEK293F and various tumor cell lines, and show that proprotein convertases can mediate the processing intracellularly as well as extracellularly. Furthermore, processed matrix metalloprotease-2 exhibits enzymatic activity that is enhanced by intermolecular autolytic cleavage. Thus, our experimental data, taken together with the broad expression of proprotein convertases, suggest that the proprotein convertase-mediated processing may be a general activation mechanism for pro-matrix metalloprotease-2 in vivo.

  4. Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro.

    PubMed

    Beek, J; Nauwynck, H; Maes, D; Van Soom, A

    2012-12-01

    In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 μM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1-TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

  5. Gold Nanoparticles Inhibit Matrix Metalloproteases without Cytotoxicity.

    PubMed

    Hashimoto, M; Sasaki, J I; Yamaguchi, S; Kawai, K; Kawakami, H; Iwasaki, Y; Imazato, S

    2015-08-01

    Nanoparticles (NPs) are currently the focus of considerable attention for dental applications; however, their biological effects have not been fully elucidated. The long-term, slow release of matrix metalloproteases (MMPs) digests collagen fibrils within resin-dentin bonds. Therefore, MMP inhibitors can prolong the durability of resin-dentin bonds. However, there have been few reports evaluating the combined effect of MMP inhibition and the cytotoxic effects of NPs for dentin bonding. The aim of this study was to evaluate MMP inhibition and cytotoxic responses to gold (AuNPs) and platinum nanoparticles (PtNPs) stabilized by polyvinylpyrrolidone (PVP) in cultured murine macrophages (RAW264) by using MMP inhibition assays, measuring cell viability and inflammatory responses (quantitative reverse transcription polymerase chain reaction [RT-qPCR]), and conducting a micromorphological analysis by fluorescence and transmission electron microscopy. Cultured RAW264 cells were exposed to metal NPs at various concentrations (1, 10, 100, and 400 µg/mL). AuNPs and PtNPs markedly inhibited MMP-8 and MMP-9 activity. Although PtNPs were cytotoxic at high concentrations (100 and 400 µg/mL), no cytotoxic effects were observed for AuNPs at any concentration. Transmission electron microscopy images showed a significant nonrandom intercellular distribution for AuNPs and PtNPs, which were mostly observed to be localized in lysosomes but not in the nucleus. RT-qPCR analysis demonstrated inflammatory responses were not induced in RAW264 cells by AuNPs or PtNPs. The cytotoxicity of nanoparticles might depend on the core metal composition and arise from a "Trojan horse" effect; thus, MMP inhibition could be attributed to the surface charge of PVP, which forms the outer coating of NPs. The negative charge of the surface coating of PVP binds to Zn(2+) from the active center of MMPs by chelate binding and results in MMP inhibition. In summary, AuNPs are attractive NPs that effectively

  6. Metalloproteases and rotator cuff disease.

    PubMed

    Del Buono, Angelo; Oliva, Francesco; Longo, Umile Giuseppe; Rodeo, Scott A; Orchard, John; Denaro, Vincenzo; Maffulli, Nicola

    2012-02-01

    The molecular changes occurring in rotator cuff tears are still unknown, but much attention has been paid to better understand the role of matrix metalloproteinases (MMP) in the development of tendinopathy. These are potent enzymes that, once activated, can completely degrade all components of the connective tissue, modify the extracellular matrix (ECM), and mediatethe development of painful tendinopathy and tendon rupture. To control the local activity of activated proteinases, the same cells produce tissue inhibitors of metalloproteinases (TIMP) that bind to the enzymes and prevent degradation. The balance between the activities of MMPs and TIMPs regulates tendon remodeling, whereas an imbalance produces a collagen dis-regulation and disturbances intendons. ADAMs (a disintegrin and metalloproteinase) are cell membrane-linked enzymes with proteolytic and cell signaling functions. ADAMTSs (ADAM with thrombospondin motifs) are secreted into the circulation, and constitute a heterogenous family of proteases with both anabolic and catabolic functions. Biologic modulation of endogenous MMP activity to basal levels may reduce pathologic tissue degradation and favorably influence healing after rotator cuff repair. Further studies are needed to better define the mechanism of action, and whether these new strategies are safe and effective in larger models.

  7. Ursolic acid reduces the metalloprotease/anti-metalloprotease imbalance in cerebral ischemia and reperfusion injury

    PubMed Central

    Wang, Yanzhe; He, Zhiyi; Deng, Shumin

    2016-01-01

    Background Activators of PPARs, particularly PPARγ, may be effective neuroprotective drugs against inflammatory responses in cerebral ischemia and reperfusion injury. Ursolic acid (UA) may act as a PPARγ agonist and serve as an anti-inflammatory agent. In this study, we used a rat middle cerebral artery occlusion and reperfusion model to examine how UA acts as a neuroprotective agent to modulate the metalloprotease/anti-metalloprotease balance. Methods The middle cerebral artery occlusion and reperfusion model (occlusion for 2 hours followed by reperfusion for 48 hours) was induced in male Sprague Dawley rats. UA was administered intragastrically 0.5, 24, and 47 hours after reperfusion. Bisphenol A diglycidyl ether (a PPARγ antagonist) was intraperitoneally administered 1, 24.5, and 47.5 hours after reperfusion. Forty-eight hours after reperfusion, neurological deficits and infarct volume were estimated. The PPARγ level and the metalloprotease/anti-metalloprotease balance were examined by Western blotting and immunohistochemistry. The activation of MAPK signaling pathways was also assessed. Results UA-treated (5, 10, or 20 mg/kg) rats showed significant improvement in neurological deficit score, infarct volume, and the number of intact neurons compared with control rats (P<0.01). Both the PPARγ protein level and the percentage of PPARγ-positive cells were increased in the UA-treated groups (P<0.01). Compared with the control group, the UA-treated groups exhibited reduced protein levels of MMP2, MMP9, and activated MAPKs (P<0.01) but an increased level of TIMP1 (P<0.01). UA exerted its protective effects in a dose-dependent manner. Co-treatment with UA and bisphenol A diglycidyl ether completely abolished the UA-induced changes in PPARγ expression; however UA continued to exert a significant but partial neuroprotective effect. Conclusion UA can act as a PPARγ agonist to improve the metalloprotease/anti-metalloprotease balance, possibly by inhibiting the

  8. Novel apigenin based small molecule that targets snake venom metalloproteases.

    PubMed

    Srinivasa, Venkatachalaiah; Sundaram, Mahalingam S; Anusha, Sebastian; Hemshekhar, Mahadevappa; Chandra Nayaka, Siddaiah; Kemparaju, Kempaiah; Basappa; Girish, Kesturu S; Rangappa, Kanchugarakoppal S

    2014-01-01

    The classical antivenom therapy has appreciably reduced snakebite mortality rate and thus is the only savior drug available. Unfortunately, it considerably fails to shield the viper bite complications like hemorrhage, local tissue degradation and necrosis responsible for severe morbidity. Moreover, the therapy is also tagged with limitations including anaphylaxis, serum sickness and poor availability. Over the last decade, snake venom metalloproteases (SVMPs) are reported to be the primary component responsible for hemorrhage and tissue degradation at bitten site. Thus, antivenom inability to offset viper venom-induced local toxicity has been a basis for an insistent search for SVMP inhibitors. Here we report the inhibitory effect of compound 5d, an apigenin based molecule against SVMPs both in silico and in vivo. Several apigenin analogues are synthesized using multicomponent Ugi reactions. Among them, compound 5d effectively abrogated Echis carinatus (EC) venom-induced local hemorrhage, tissue necrosis and myotoxicity in a dose dependant fashion. The histopathological study further conferred effective inhibition of basement membrane degradation, and accumulation of inflammatory leucocytes at the site of EC venom inoculation. The compound also protected EC venom-induced fibrin and fibrinogen degradation. The molecular docking of compound 5d and bothropasin demonstrated the direct interaction of hydroxyl group of compound with Glu146 present in hydrophobic pocket of active site and does not chelate Zn2+. Hence, it is concluded that compound 5d could be a potent agent in viper bite management.

  9. Catalytic Domain Architecture of Metzincin Metalloproteases*

    PubMed Central

    Gomis-Rüth, F. Xavier

    2009-01-01

    Metalloproteases cleave proteins and peptides, and deregulation of their function leads to pathology. An understanding of their structure and mechanisms of action is necessary to the development of strategies for their regulation. Among metallopeptidases are the metzincins, which are mostly multidomain proteins with ∼130–260-residue globular catalytic domains showing a common core architecture characterized by a long zinc-binding consensus motif, HEXXHXXGXX(H/D), and a methionine-containing Met-turn. Metzincins participate in unspecific protein degradation such as digestion of intake proteins and tissue development, maintenance, and remodeling, but they are also involved in highly specific cleavage events to activate or inactivate themselves or other (pro)enzymes and bioactive peptides. Metzincins are subdivided into families, and seven such families have been analyzed at the structural level: the astacins, ADAMs/adamalysins/reprolysins, serralysins, matrix metalloproteinases, snapalysins, leishmanolysins, and pappalysins. These families are reviewed from a structural point of view. PMID:19201757

  10. Metalloprotease ADAM10 Is Required for Notch1 Site 2 Cleavage*

    PubMed Central

    van Tetering, Geert; van Diest, Paul; Verlaan, Ingrid; van der Wall, Elsken; Kopan, Raphael; Vooijs, Marc

    2009-01-01

    Notch signaling is controlled by ligand binding, which unfolds a negative control region to induce proteolytic cleavage of the receptor. First, a membrane-proximal cleavage is executed by a metalloprotease, removing the extracellular domain. This allows γ-secretase to execute a second cleavage within the Notch transmembrane domain, which releases the intracellular domain to enter the nucleus. Here we show that the ADAM10 metalloprotease Kuzbanian, but not ADAM17/tumor necrosis factor α-converting enzyme, plays an essential role in executing ligand-induced extracellular cleavage at site 2 (S2) in cells and localizes this step to the plasma membrane. Importantly, genetic or pharmacological inhibition of metalloproteases still allowed extracellular cleavage of Notch, indicating the presence of unknown proteases with the ability to cleave at S2. Gain of function mutations identified in human cancers and in model organisms that map to the negative control region alleviate the requirement for ligand binding for extracellular cleavage to occur. Because cancer-causing Notch1 mutations also depend on (rate-limiting) S2 proteolysis, the identity of these alternative proteases has important implications for understanding Notch activation in normal and cancer cells. PMID:19726682

  11. "Click" synthesis of small molecule probes for activity-based fingerprinting of matrix metalloproteases.

    PubMed

    Wang, Jun; Uttamchandani, Mahesh; Li, Junqi; Hu, Mingyu; Yao, Shao Q

    2006-09-28

    By using "Click Chemistry", we achieved the facile synthesis of various affinity-based hydroxamate probes that enable generation of activity-based fingerprints of a variety of metalloproteases, including matrix metalloproteases (MMPs), in proteomics experiments.

  12. Maturation pathway of metalloprotease produced by Aeromonas sobria.

    PubMed

    Takahashi, Eizo; Fujii, Yoshio; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Nair, Gopinath Balakrish; Takeda, Yoshifumi; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2010-10-01

    Previously, we cloned the metalloprotease gene of Aeromonas sobria (amp) and determined its nucleotide sequence (GenBank accession number DQ784565). The protease is composed of 591 amino acid residues. In this study, we purified the mature metalloprotease from the culture supernatant of A. sobria and determined the amino terminal sequence and molecular size of AMP. In addition, we examined the production of AMP diachronically and found that AMP emerges outside of the cell as an intermediate composed of mature and propeptide regions. Subsequently, we determined that the N-terminal amino acid sequence of the intermediate and found that the sequence is identical to that of the mature metalloprotease. This means that the intermediate is composed of a mature AMP region and a C-terminal propeptide. The cross culture experiment of mutants of metalloprotease and serine protease of A. sobria on skim milk agar medium indicates that the intermediate released outside of the cell is inactive and that serine protease produced by A. sobria accelerates the conversion of the intermediate from the inactive to the active form.

  13. Comparative expression and tissue distribution analyses of astacin-like squid metalloprotease in squid and cuttlefish.

    PubMed

    Kanzawa, Nobuyuki; Ogawa, Takuya; Asakura, Masanori; Okiyama, Keisuke; Honda, Michiyo; Tsuchiya, Takahide

    2008-01-01

    Astacin-like squid metalloprotease (ALSM) is a member of the astacin family of metalloproteases. In the present study, we investigated the expression and tissue distribution of ALSM in bigfin reef squid (Sepioteuthis lessoniana) and golden cuttlefish (Sepia esculenta). Myosin heavy chain hydrolysis tests showed ALSM-I-like activity in both species. We isolated partial cDNA clones showing high sequence similarity to ALSM-I and -III, suggesting that ALSM is common to squid and cuttlefish. Phylogenetic analysis showed that ALSMs are classified into two clades: ALSM-I forms one clade, and ALSM-II and -III form the other. ALSM was expressed in several tissues in bigfin reef squid, though expression was confined to the liver in cuttlefish. ALSMs are distributed in digestive organs but not in mantle muscle of squid and cuttlefish. Immunofluorescence analysis further showed that cellular localization of ALSM is evident not only in hepatic cells but also in pancreatic cells of bigfin reef squid. Thus, ALSM is commonly expressed in squid and cuttlefish, but its expression levels and distribution are distinct.

  14. Electrophysiological Characterization of the Antarease Metalloprotease from Tityus serrulatus Venom

    PubMed Central

    Zornetta, Irene; Scorzeto, Michele; Mendes Dos Reis, Pablo Victor; De Lima, Maria E.; Montecucco, Cesare; Megighian, Aram; Rossetto, Ornella

    2017-01-01

    Scorpions are among the oldest venomous living organisms and the family Buthidae is the largest and most medically relevant one. Scorpion venoms include many toxic peptides, but recently, a metalloprotease from Tityus serrulatus called antarease was reported to be capable of cleaving VAMP2, a protein involved in the neuroparalytic syndromes of tetanus and botulism. We have produced antarease and an inactive metalloprotease mutant in a recombinant form and analyzed their enzymatic activity on recombinant VAMP2 in vitro and on mammalian and insect neuromuscular junction. The purified recombinant antarease paralyzed the neuromuscular junctions of mice and of Drosophila melanogaster whilst the mutant was inactive. We were unable to demonstrate any cleavage of VAMP2 under conditions which leads to VAMP proteolysis by botulinum neurotoxin type B. Antarease caused a reduced release probability, mainly due to defects upstream of the synaptic vesicles fusion process. Paired pulse experiments indicate that antarease might proteolytically inactivate a voltage-gated calcium channel. PMID:28264432

  15. A Disintegrin and Metalloprotease (ADAM): Historical Overview of Their Functions

    PubMed Central

    Giebeler, Nives; Zigrino, Paola

    2016-01-01

    Since the discovery of the first disintegrin protein from snake venom and the following identification of a mammalian membrane-anchored metalloprotease-disintegrin implicated in fertilization, almost three decades of studies have identified additional members of these families and several biochemical mechanisms regulating their expression and activity in the cell. Most importantly, new in vivo functions have been recognized for these proteins including cell partitioning during development, modulation of inflammatory reactions, and development of cancers. In this review, we will overview the a disintegrin and metalloprotease (ADAM) family of proteases highlighting some of the major research achievements in the analysis of ADAMs’ function that have underscored the importance of these proteins in physiological and pathological processes over the years. PMID:27120619

  16. Neprilysin II: A putative novel metalloprotease and its isoforms in CNS and testis.

    PubMed

    Ouimet, T; Facchinetti, P; Rose, C; Bonhomme, M C; Gros, C; Schwartz, J C; Tanja, O

    2000-05-19

    Metalloproteases of the M13 subfamily, comprising namely neprylisin (NEP) and endothelin-converting enzyme (ECE), are involved in the metabolism of various neuronal and hormonal peptides, and inhibitors thereof have already led to therapeutically useful agents. Using homology cloning, we have identified a new member of this family in rat tissues. It is a glycosylated, type II integral membrane protein of 774 amino acids, containing a zinc-binding consensus motif, highly homologous to NEP and, therefore, designated NEPII. We have characterized multiple splice variants of NEPII mRNA with distinct expression patterns in brain regions, pituitary and testis. In situ hybridization of testis, where levels of the NEPII gene transcript are the highest, reveals a localization within round spermatids. In brain, NEPII is expressed heterogeneously among several neuronal populations and according to a pattern grossly complementary to that of NEP.

  17. The enterotoxin of Bacteroides fragilis is a metalloprotease.

    PubMed Central

    Moncrief, J S; Obiso, R; Barroso, L A; Kling, J J; Wright, R L; Van Tassell, R L; Lyerly, D M; Wilkins, T D

    1995-01-01

    During the past decade, strains of Bacteroides fragilis that produce an enterotoxin have been implicated in diarrheal disease in animals and humans. The extracellular enterotoxin has been purified and characterized as a single polypeptide (M(r), approximately 20,000). Single specific primer-PCR was used to clone a portion of the B. fragilis enterotoxin gene. The recombinant protein expressed by the cloned gene fragment reacted with monospecific antibodies to B. fragilis enterotoxin by enzyme-linked immunosorbent assay and immunoblot analysis. The deduced amino acid sequence revealed a signature zinc-binding consensus motif (HEXXHXXGXXH/Met-turn) characteristic of metalloproteases termed metzincins. Sequence comparisons showed close identity to matrix metalloproteases (e.g., human fibroblast collagenase) within the zinc-binding and Met-turn region. Purified enterotoxin contained 1 g-atom of Zn2+ per molecule and hydrolyzed gelatin, azocoll, actin, tropomyosin, and fibrinogen. The enterotoxin also underwent autodigestion. The N-terminal amino acid sequences of two autodigestion products were identical to the deduced amino acid sequence of the recombinant enterotoxin and revealed cleavage at Cys-Leu and Ser-Leu peptide bonds. Gelatinase (type IV collagenase) activity comigrated with the toxin when analyzed by gel fractionation and zymography, indicating that protease activity is due to the enterotoxin and not to a contaminating protease(s). Optimal proteolytic activity occurred at 37 degrees C and pH 6.5. Primary proteolytic cleavage sites in actin were identified, revealing cleavage at Gly-Met and Thr-Leu peptide bonds. Enzymatic activity was inhibited by metal chelators but not by inhibitors of other classes of proteases. Additionally, cytotoxic activity of the enterotoxin on human carcinoma HT-29 cells was inhibited by acetoxymethyl ester EDTA. The metalloprotease activity of the enterotoxin suggests a possible mechanism for enterotoxicity and may have additional

  18. Functional analysis of a breast cancer-associated mutation in the intracellular domain of the metalloprotease ADAM12.

    PubMed

    Stautz, Dorte; Wewer, Ulla M; Kveiborg, Marie

    2012-01-01

    A recently identified breast cancer-associated mutation in the metalloprotease ADAM12 alters a potential dileucine trafficking signal, which could affect protein processing and cellular localization. ADAM12 belongs to the group of A Disintegrin And Metalloproteases (ADAMs), which are typically membrane-associated proteins involved in ectodomain shedding, cell-adhesion, and signaling. ADAM12 as well as several members of the ADAM family are over-expressed in various cancers, correlating with disease stage. Three breast cancer-associated somatic mutations were previously identified in ADAM12, and two of these, one in the metalloprotease domain and another in the disintegrin domain, were investigated and found to result in protein misfolding, retention in the secretory pathway, and failure of zymogen maturation. The third mutation, p.L792F in the ADAM12 cytoplasmic tail, was not investigated, but is potentially significant given its location within a di-leucine motif, which is recognized as a potential cellular trafficking signal. The present study was motivated both by the potential relevance of this documented mutation to cancer, as well as for determining the role of the di-leucine motif in ADAM12 trafficking. Expression of ADAM12 p.L792F in mammalian cells demonstrated quantitatively similar expression levels and zymogen maturation as wild-type (WT) ADAM12, as well as comparable cellular localizations. A cell surface biotinylation assay demonstrated that cell surface levels of ADAM12 WT and ADAM12 p.L792F were similar and that internalization of the mutant occurred at the same rate and extent as for ADAM12 WT. Moreover, functional analysis revealed no differences in cell proliferation or ectodomain shedding of epidermal growth factor (EGF), a known ADAM12 substrate between WT and mutant ADAM12. These data suggest that the ADAM12 p.L792F mutation is unlikely to be a driver (cancer causing)-mutation in breast cancer.

  19. A novel synthetic quinolinone inhibitor presents proteolytic and hemorrhagic inhibitory activities against snake venom metalloproteases.

    PubMed

    Baraldi, Patrícia T; Magro, Angelo J; Matioli, Fábio F; Marcussi, Silvana; Lemke, Ney; Calderon, Leonardo A; Stábeli, Rodrigo G; Soares, Andreimar M; Correa, Arlene G; Fontes, Marcos R M

    2016-02-01

    Metalloproteases play a fundamental role in snake venom envenomation inducing hemorrhagic, fibrigen(ogen)olytic and myotoxic effects in their victims. Several snake venoms, such as those from the Bothrops genus, present important local effects which are not efficiently neutralized by conventional serum therapy. Consequently, these accidents may result in permanent sequelae and disability, creating economic and social problems, especially in developing countries, leading the attention of the World Health Organization that considered ophidic envenomations a neglected tropical disease. Aiming to produce an efficient inhibitor against bothropic venoms, we synthesized different molecules classified as quinolinones - a group of low-toxic chemical compounds widely used as antibacterial and antimycobacterial drugs - and tested their inhibitory properties against hemorrhage caused by bothropic venoms. The results from this initial screening indicated the molecule 2-hydroxymethyl-6-methoxy-1,4-dihydro-4-quinolinone (Q8) was the most effective antihemorrhagic compound among all of the assayed synthetic quinolinones. Other in vitro and in vivo experiments showed this novel compound was able to inhibit significantly the hemorrhagic and/or proteolytic activities of bothropic crude venoms and isolated snake venom metalloproteases (SVMPs) even at lower concentrations. Docking and molecular dynamic simulations were also performed to get insights into the structural basis of Q8 inhibitory mechanism against proteolytic and hemorrhagic SVMPs. These structural studies demonstrated that Q8 may form a stable complex with SVMPs, impairing the access of substrates to the active sites of these toxins. Therefore, both experimental and structural data indicate that Q8 compound is an interesting candidate for antiophidic therapy, particularly for the treatment of the hemorrhagic and necrotic effects induced by bothropic venoms.

  20. Genetic inactivation of ADAMTS15 metalloprotease in human colorectal cancer.

    PubMed

    Viloria, Cristina G; Obaya, Alvaro J; Moncada-Pazos, Angela; Llamazares, María; Astudillo, Aurora; Capellá, Gabriel; Cal, Santiago; López-Otín, Carlos

    2009-06-01

    Matrix metalloproteinases have been traditionally linked to cancer dissemination through their ability to degrade most extracellular matrix components, thus facilitating invasion and metastasis of tumor cells. However, recent functional studies have revealed that some metalloproteases, including several members of the ADAMTS family, also exhibit tumor suppressor properties. In particular, ADAMTS1, ADAMTS9, and ADAMTS18 have been found to be epigenetically silenced in malignant tumors of different sources, suggesting that they may function as tumor suppressor genes. Herein, we show that ADAMTS15 is genetically inactivated in colon cancer. We have performed a mutational analysis of the ADAMTS15 gene in human colorectal carcinomas, with the finding of four mutations in 50 primary tumors and 6 colorectal cancer cell lines. Moreover, functional in vitro and in vivo studies using HCT-116 and SW-620 colorectal cancer cells and severe combined immunodeficient mice have revealed that ADAMTS15 restrains tumor growth and invasion. Furthermore, the presence of ADAMTS15 in human colorectal cancer samples showed a negative correlation with the histopathologic differentiation grade of the corresponding tumors. Collectively, these results provide evidence that extracellular proteases, including ADAMTS15, may be targets of inactivating mutations in human cancer and further validate the concept that secreted metalloproteases may show tumor suppressor properties.

  1. Control of Entamoeba histolytica adherence involves metallosurface protease 1, an M8 family surface metalloprotease with homology to leishmanolysin.

    PubMed

    Teixeira, Jose E; Sateriale, Adam; Bessoff, Kovi E; Huston, Christopher D

    2012-06-01

    Invasive amebiasis due to Entamoeba histolytica infection is an important cause of morbidity in developing countries. The E. histolytica genome contains two homologues to the metalloprotease leishmanolysin gene, Entamoeba histolytica MSP-1 (EhMSP-1) and EhMSP-2, while the commensal ameba Entamoeba dispar has lost EhMSP-1. In this study, we sought to characterize E. histolytica metallosurface protease 1 (EhMSP-1). Using immunoprecipitation and a model substrate, we found that EhMSP-1 was a functional metalloprotease. Confocal microscopy and flow cytometry revealed that EhMSP-1 localized to the cell surface and revealed the existence of distinct, nonclonal trophozoite populations with high and low EhMSP-1 surface abundance that became synchronized following serum starvation. Phenotypic assays were performed after silencing EhMSP-1. Adherence of EhMSP-1-deficient trophozoites to tissue culture cell monolayers was more than five times greater than that of control amebas, but surface staining of several antigens, including the galactose adherence lectin, was unchanged. EhMSP-1 silencing similarly increased adherence to both viable and apoptotic Jurkat lymphocytes. Tissue culture cell monolayer destruction was reduced by EhMSP-1 silencing, although it was blocked almost completely by inhibiting cysteine proteases. Consistent with a primary defect in regulation of amebic adherence, EhMSP-1 silencing also resulted in reduced mobility on tissue culture cell monolayers and in increased phagocytosis. In conclusion, EhMSP-1 was shown to be a surface metalloprotease involved in regulation of amebic adherence, with additional effects on cell motility, cell monolayer destruction, and phagocytosis.

  2. The Metalloprotease Neprilysin Degrades and Inactivates Apelin Peptides.

    PubMed

    McKinnie, Shaun M K; Fischer, Conrad; Tran, Kelvin M H; Wang, Wang; Mosquera, Fabricio; Oudit, Gavin Y; Vederas, John C

    2016-08-17

    The apelinergic system is a mammalian peptide hormone network with key physiological roles. Apelin isoforms and analogues are believed to be promising therapeutics for cardiovascular disease. Despite extensive studies on apelin-13 degradation patterns, only one protease, angiotensin-converting enzyme 2 (ACE2), had been implicated in its physiological regulation. Through use of a peptide-based fluorescent probe, we identified the metalloprotease neprilysin (NEP, a target for Entresto used in treatment of heart failure) as an enzyme that cleaves apelin isoforms. In vitro NEP proteolysis generated fragments that lacked the ability to bind to the apelin receptor, thereby making NEP the first protease to fully inactivate apelin. The involvement of NEP in the apelinergic system contributes to the understanding of its role in cardiovascular physiology.

  3. ADM-1, a protein with metalloprotease- and disintegrin-like domains, is expressed in syncytial organs, sperm, and sheath cells of sensory organs in Caenorhabditis elegans.

    PubMed Central

    Podbilewicz, B

    1996-01-01

    A search was carried out for homologues of possible fusogenic proteins to study their function in a genetically tractable animal. The isolation, molecular, and cellular characterization of the Caenorhabditis elegans adm-1 gene (a disintegrin and metalloprotease domain) are described. A glycoprotein analogous to viral fusion proteins has been identified on the surface of guinea pig sperm (PH-30/fertilin) and is implicated in sperm-egg fusion. adm-1 is the first reported invertebrate gene related to PH-30 and a family of proteins containing snake venom disintegrin- and metalloprotease-like domains. ADM-1 shows a domain organization identical to PH-30. It contains prepro, metalloprotease, disintegrin, cysteine rich with putative fusion peptide, epidermal growth factor-like repeat, transmembrane, and cytoplasmic domains. Antibodies which recognize ADM-1 protein in immunoblots were generated. Using immunofluorescence and in situ hybridization, the products of adm-1 have been detected in specific cells during different stages of development. The localization of ADM-1 to the plasma membrane of embryonic cells and to the sheath cells of sensory organs suggests a function in cell adhesion. ADM-1 expression in the hypodermis, pharynx, vulva, and mature sperm is consistent with a putative role in somatic and gamete cell fusions. Images PMID:8970152

  4. The emerging role of matrix metalloproteases of the ADAM family in male germ cell apoptosis

    PubMed Central

    Urriola-Muñoz, Paulina; Lagos-Cabré, Raúl

    2011-01-01

    Constitutive germ cell apoptosis during mammalian spermatogenesis is a key process for controlling sperm output and to eliminate damaged or unwanted cells. An increase or decrease in the apoptosis rate has deleterious consequences and leads to low sperm production. Apoptosis in spermatogenesis has been widely studied, but the mechanism by which it is induced under physiological or pathological conditions has not been clarified. We have recently identified the metalloprotease ADAM17 (TACE) as a putative physiological inducer of germ cell apoptosis. The mechanisms involved in regulating the shedding of the ADAM17 extracellular domain are still far from being understood, although they are important in order to understand cell-cell communications. Here, we review the available data regarding apoptosis during mammalian spermatogenesis and the localization of ADAM proteins in the male reproductive tract. We propose an integrative working model where ADAM17, p38 MAPK, protein kinase C (PKC) and the tyrosine kinase c-Abl participate in the physiological signalling cascade inducing apoptosis in germ cells. In our model, we also propose a role for the Sertoli cell in regulating the Fas/FasL system in order to induce the extrinsic pathway of apoptosis in germ cells. This working model could be applied to further understand constitutive apoptosis in spermatogenesis and in pathological conditions (e.g., varicocele) or following environmental toxicants exposure (e.g., genotoxicity or xenoestrogens). PMID:22319668

  5. ADAMTS metalloproteases generate active versican fragments that regulate interdigital web regression

    PubMed Central

    McCulloch, Daniel R.; Nelson, Courtney M.; Dixon, Laura J.; Silver, Debra L.; Wylie, James D.; Lindner, Volkhard; Sasaki, Takako; Cooley, Marion A.; Argraves, W. Scott; Apte, Suneel S.

    2009-01-01

    We show that combinatorial mouse alleles for the secreted metalloproteases Adamts5, Adamts20 (bt), and Adamts9 result in fully penetrant soft-tissue syndactyly. Interdigital webs in Adamts5−/−; bt/bt mice had reduced apoptosis and decreased cleavage of the proteoglycan versican; however, the BMP-FGF axis, which regulates interdigital apoptosis was unaffected. BMP4 induced apoptosis, but without concomitant versican proteolysis. Haploinsufficiency of either Vcan or Fbln1, a co-factor for versican processing by ADAMTS5, led to highly penetrant syndactyly in bt mice, suggesting that cleaved versican was essential for web regression. The local application of an amino-terminal versican fragment corresponding to ADAMTS-processed versican, induced cell death in Adamts5−/−; bt/bt webs. Thus, ADAMTS proteases cooperatively maintain versican proteolysis above a required threshold to create a permissive environment for apoptosis. The data highlight the developmental significance of proteolytic action on the ECM, not only as a clearance mechanism, but also as a means to generate bioactive versican fragments. PMID:19922873

  6. A fluorescent peptide substrate for the surface metalloprotease of Leishmania.

    PubMed

    Bouvier, J; Schneider, P; Malcolm, B

    1993-03-01

    A fluorescent oligopeptide substrate for the promastigote surface protease (PSP) of Leishmania was designed using the data reported for the substrate specificity of the enzyme (Bouvier, J., Schneider, P., Etges, R. J., and Bordier, C. 1990. Biochemistry 29, 10113-10119). The indole fluorescence of the tryptophan residue was efficiently quenched through resonance energy transfer by an N-terminal dansyl group located five amino acid residues away. The heptapeptide, dansyl-A-Y-L-K-K-W-V-NH2, was cleaved by PSP between the tyrosine and leucine residues with a kcat/Km ratio of 8.8 x 10(6) M-1sec-1. Hydrolysis by the enzyme results in a time-dependent increase of fluorescence intensity of 3.7-fold. Assays can be designed based on the tryptophan fluorescence at 360 nm or by individual product analyses using thin-layer chromatography. The synthetic substrate is readily cleaved by the metalloprotease at the surface of fixed promastigotes. The specificity and sensitivity of such internally quenched fluorescent peptide substrate will facilitate the identification of novel inhibitors for the enzyme and aid in detailed studies on its enzymology.

  7. Time-dependent botulinum neurotoxin serotype A metalloprotease inhibitors

    PubMed Central

    Cardinale, Steven C.; Butler, Michelle M.; Pai, Ramdas; Nuss, Jonathan E.; Peet, Norton P.; Bavari, Sina; Bowlin, Terry L.

    2011-01-01

    Botulinum neurotoxins (BoNTs) are the most lethal of biological substances, and are categorized as class A biothreat agents by the Centers for Disease Control and Prevention. There are currently no drugs to treat the deadly flaccid paralysis resulting from BoNT intoxication. Among the seven BoNT serotypes, the development of therapeutics to counter BoNT/A is a priority (due to its long half-life in the neuronal cytosol and its ease of production). In this regard, the BoNT/A enzyme light chain (LC) component, a zinc metalloprotease responsible for the intracellular cleavage of synaptosomal-associated protein of 25 kDa, is a desirable target for developing post-BoNT/A intoxication rescue therapeutics. In an earlier study, we reported the high throughput screening of a library containing 70,000 compounds, and uncovered a novel class of benzimidazole acrylonitrile-based BoNT/A LC inhibitors. Herein, we present both structure-activity relationships and a proposed mechanism of action for this novel inhibitor chemotype. PMID:22082667

  8. Immunoprotective potential of a Rhipicephalus (Boophilus) microplus metalloprotease.

    PubMed

    Ali, Abid; Parizi, Luís Fernando; Guizzo, Melina Garcia; Tirloni, Lucas; Seixas, Adriana; Vaz, Itabajara da Silva; Termignoni, Carlos

    2015-01-15

    Ticks have serious impacts on animal and human health, causing significant economic losses in cattle breeding. Besides damage due to the hematophagous behavior, they transmit several pathogens. Low cost and environmental safety have made vaccines a promising alternative control method against tick infestation. Metalloproteases (MPs) have been shown to be essential for diverse biological functions in hematophagous organisms, inhibiting blood clotting, degrading extracellular matrix proteins, and inhibiting host tissue repair via anti-angiogenic activity. In this study, we analyzed the immunoprotective potential of a recombinant MP against Rhipicephalus (Boophilus) microplus infestation. First, a cDNA encoding R. microplus amino acids sequence with highly conserved regions of the metzincin (reprolysin) group of MP was identified (BrRm-MP4). After expression and purification, recombinant BrRm-MP4 was used as a vaccinal antigen against R. microplus infestation in cattle (Bos taurus taurus). All vaccinated bovines developed immune response to the antigen, resulting in increased antibody level throughout the immunization protocol. Immunization with rBrRm-MP4 reduced tick feeding success, decreasing the number of engorged females and their reproduction potential, representing a 60% overall protection. These results show that rBrRm-MP4 provides protection against tick infestation, placing it is a potential candidate for an anti-tick vaccine.

  9. Recent advances in the design of matrix metalloprotease inhibitors.

    PubMed

    Matter, Hans; Schudok, Manfred

    2004-07-01

    Inhibition of matrix metalloproteases (MMPs) for the treatment of diseases, such as cancer, arthritis and other diseases associated with tissue remodeling, has become an area of intense interest in the pharmaceutical industry in recent years. Despite tremendous efforts over the last decade to explore individual members of this target family, along with multiple inhibitor classes, simple and effective drugs for inhibiting individual MMPs have not yet emerged. This review highlights the major developments in research into MMPs and their inhibitors, from the recent medicinal chemistry literature, with a focus on structure-based design, selectivity and pharmacokinetic (PK) properties. The increasing availability of high-resolution X-ray crystal structures for many members of this protein family makes MMPs ideally suited for structure-based design approaches, which are now routinely used in this area. The most challenging aspect of lead optimization for MMP inhibitors is in finding candidates having acceptable pharmacological, PK and selectivity profiles. Clinical trials in cancer giving disappointing results have led to discussions on how to gain adequate MMP selectivity in order to minimize side effects. Unfortunately, careful analysis of X-ray crystal structures has not suggested any simple solutions. These areas collectively constitute the main challenges in the current search for orally available MMP inhibitors, and will be discussed in this review.

  10. The plastid metalloprotease FtsH6 and small heat shock protein HSP21 jointly regulate thermomemory in Arabidopsis

    PubMed Central

    Sedaghatmehr, Mastoureh; Mueller-Roeber, Bernd; Balazadeh, Salma

    2016-01-01

    Acquired tolerance to heat stress is an increased resistance to elevated temperature following a prior exposure to heat. The maintenance of acquired thermotolerance in the absence of intervening stress is called ‘thermomemory' but the mechanistic basis for this memory is not well defined. Here we show that Arabidopsis HSP21, a plastidial small heat shock protein that rapidly accumulates after heat stress and remains abundant during the thermomemory phase, is a crucial component of thermomemory. Sustained memory requires that HSP21 levels remain high. Through pharmacological interrogation and transcriptome profiling, we show that the plastid-localized metalloprotease FtsH6 regulates HSP21 abundance. Lack of a functional FtsH6 protein promotes HSP21 accumulation during the later stages of thermomemory and increases thermomemory capacity. Our results thus reveal the presence of a plastidial FtsH6–HSP21 control module for thermomemory in plants. PMID:27561243

  11. Purification and Characterization of a Serratia marcescens Metalloprotease

    PubMed Central

    Lyerly, David; Kreger, Arnold

    1979-01-01

    An extracellular, nonelastolytic, neutral metalloprotease of Serratia marcescens was purified by sequential ammonium sulfate precipitation, hydroxyapatite adsorption chromatography, flat-bed isoelectric focusing, and Sephadex G-100 gel filtration. The protease preparation had a 280/260 nm absorbance ratio of 1.8, was free of detectable amounts of endotoxin, carbohydrate, phosphorus, and other known extracellular enzymes of S. marcescens, and was homogeneous by Ouchterlony double immunodiffusion and Grabar-Williams immunoelectrophoresis. Crossed immunoelectrophoresis, thin-layer electrofocusing in polyacrylamide gel, and polyacrylamide disc gel electrophoresis showed three to four closely migrating, Coomassie blue-staining components in the protease preparation. However, zymogram analyses of the patterns showed that protease activity was associated with each component and that the protease was, therefore, microheterogeneous. The isoelectric point and sedimentation coefficient of the protease were approximately 5.3 to 5.4 and 4.2S, respectively, and the molecular weight estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by gel filtration was approximately 52,500 and 44,000, respectively. The pH optimum range, with azocasein as the substrate, was 5.5 to 7.5. The enzyme contained a high percentage of acidic amino acids, no cysteine, and 1 g-atom of Zn2+ and 7 g-atoms of Ca2+ per mol. Various heavy metal ions and chelating agents and heating at 60°C for 15 min inactivated the enzyme. Intracorneal, intratracheal, and intradermal administration of the protease into rabbits elicited rapid and extensive tissue damage. The minimum lethal intravenous dose for mice was approximately 17 mg/kg of body weight. Images PMID:37166

  12. Mycobacterium tuberculosis Zinc Metalloprotease-1 Assists Mycobacterial Dissemination in Zebrafish

    PubMed Central

    Vemula, Mani H.; Medisetti, Raghavender; Ganji, Rakesh; Jakkala, Kiran; Sankati, Swetha; Chatti, Kiranam; Banerjee, Sharmistha

    2016-01-01

    Zinc metalloprotease-1 (Zmp1) from Mycobacterium tuberculosis (M.tb), the tuberculosis (TB) causing bacillus, is a virulence factor involved in inflammasome inactivation and phagosome maturation arrest. We earlier reported that Zmp1 was secreted under granuloma-like stress conditions, induced Th2 cytokine microenvironment and was highly immunogenic in TB patients as evident from high anti-Zmp1 antibody titers in their sera. In this study, we deciphered a new physiological role of Zmp1 in mycobacterial dissemination. Exogenous treatment of THP-1 cells with 500 nM and 1 μM of recombinant Zmp1 (rZmp1) resulted in necrotic cell death. Apart from inducing secretion of necrotic cytokines, TNFα, IL-6, and IL-1β, it also induced the release of chemotactic chemokines, MCP-1, MIP-1β, and IL-8, suggesting its likely function in cell migration and mycobacterial dissemination. This was confirmed by Gap closure and Boyden chamber assays, where Zmp1 treated CHO or THP-1 cells showed ∼2 fold increased cell migration compared to the untreated cells. Additionally, Zebrafish-M. marinum based host–pathogen model was used to study mycobacterial dissemination in vivo. Td-Tomato labeled M. marinum (TdM. marinum) when injected with rZmp1 showed increased dissemination to tail region from the site of injection as compared to the untreated control fish in a dose-dependent manner. Summing up these observations along with the earlier reports, we propose that Zmp1, a multi-faceted protein, when released by mycobacteria in granuloma, may lead to necrotic cell damage and release of chemotactic chemokines by surrounding infected macrophages, attracting new immune cells, which in turn may lead to fresh cellular infections, thus assisting mycobacterial dissemination. PMID:27621726

  13. Wound bed preparation 2014 update: management of critical colonization with a gentian violet and methylene blue absorbent antibacterial dressing and elevated levels of matrix metalloproteases with an ovine collagen extracellular matrix dressing.

    PubMed

    Sibbald, R Gary; Ovington, Liza G; Ayello, Elizabeth A; Goodman, Laurie; Elliott, James A

    2014-03-01

    Wound bed preparation (WBP) is a paradigm for holistic patient care that includes treatment of the cause along with patient-centered concerns before optimizing the components of local wound care (debridement, infection/inflammation, moisture balance, and, when required, the edge effect). This review incorporates a methylene blue and gentian violet bound foam dressing for critical colonization and an ovine collagen extracellular matrix dressing for reduction of elevated levels of matrix metalloproteases into the WBP paradigm.

  14. Helicobacter pylori and gastritis: the role of extracellular matrix metalloproteases, their inhibitors, and the disintegrins and metalloproteases--a systematic literature review.

    PubMed

    Sampieri, Clara L

    2013-10-01

    Helicobacter pylori (H. pylori) is the etiologic agent of gastritis; it has been estimated that 50 % of the world's population could be infected by this bacteria. Gastritis may progress to chronic atrophic gastritis, a condition associated with the development of gastric cancer (GC). Several matrix metalloproteases (MMP) and tissue inhibitors of MMPs (TIMP) as well as disintegrins and metalloproteases (ADAM) have been reported as being involved in gastritis. Among other processes, these protein families participate in remodeling the extracellular matrix, cell signaling, immune response, angiogenesis, inflammation and epithelial mesenchymal transition. This systematic review analyzes the scientific evidence surrounding the relationship between members of the MMP, TIMP and ADAM families and infection by H. pylori in gastritis, considering both in vitro and in vivo studies. Given the potential clinical value of certain members of the MMP, TIMP and ADAM families as molecular markers in gastritis and the association of gastritis with GC, the need for further study is highlighted.

  15. A Putative Chloroplast Thylakoid Metalloprotease VIRESCENT3 Regulates Chloroplast Development in Arabidopsis thaliana*

    PubMed Central

    Qi, Yafei; Liu, Xiayan; Liang, Shuang; Wang, Rui; Li, Yuanfeng; Zhao, Jun; Shao, Jingxia; An, Lijun; Yu, Fei

    2016-01-01

    The chloroplast is the site of photosynthesis and many other essential plant metabolic processes, and chloroplast development is an integral part of plant growth and development. Mutants defective in chloroplast development can display various color phenotypes including the intriguing virescence phenotype, which shows yellow/white coloration at the leaf base and greening toward the leaf tip. Through large scale genetic screens, we identified a series of new virescent mutants including virescent3-1 (vir3-1), vir4-1, and vir5-1 in Arabidopsis thaliana. We showed that VIR3 encodes a putative chloroplast metalloprotease by map-based cloning. Through site-directed mutagenesis, we showed that the conserved histidine 235 residue in the zinc binding motif HEAGH of VIR3 is indispensable for VIR3 accumulation in the chloroplast. The chloroplast localization of VIR3 was confirmed by the transient expression of VIR3-GFP in leaf protoplasts. Furthermore, taking advantage of transgenic lines expressing VIR3-FLAG, we demonstrated that VIR3 is an intrinsic thylakoid membrane protein that mainly resides in the stromal lamellae. Moreover, topology analysis using transgenic lines expressing a dual epitope-tagged VIR3 indicated that both the N and C termini of VIR3 are located in the stroma, and the catalytic domain of VIR3 is probably facing the stroma. Blue native gel analysis indicated that VIR3 is likely present as a monomer or part of a small complex in the thylakoid membrane. This work not only implicates VIR3 as a new factor involved in early chloroplast development but also provides more insight into the roles of chloroplast proteases in chloroplast biogenesis. PMID:26702056

  16. Metalloprotease Peptide Inhibitors: A Semester-Long Organic Synthetic Research Project for the Introductory Laboratory Course

    ERIC Educational Resources Information Center

    Pontrello, Jason K.

    2015-01-01

    A semester-long research project to synthesize unique compounds designed after published metalloprotease peptide inhibitors is presented. The research project encompasses a set of nine organic chemistry reactions traditionally taught in the second semester lab course, and the procedures are derived from scientific literature. The two principle…

  17. Activation of the human keratinocyte B1 bradykinin receptor induces expression and secretion of metalloproteases 2 and 9 by transactivation of epidermal growth factor receptor.

    PubMed

    Matus, Carola E; Ehrenfeld, Pamela; Pavicic, Francisca; González, Carlos B; Concha, Miguel; Bhoola, Kanti D; Burgos, Rafael A; Figueroa, Carlos D

    2016-09-01

    The B1 bradykinin receptor (BDKRB1) is a component of the kinin cascade localized in the human skin. Some of the effects produced by stimulation of BDKRB1 depend on transactivation of epidermal growth factor receptor (EGFR), but the mechanisms involved in this process have not been clarified yet. The primary purpose of this study was to determine the effect of a BDKRB1 agonist on wound healing in a mouse model and the migration and secretion of metalloproteases 2 and 9 from human HaCaT keratinocytes and delineate the signalling pathways that triggered their secretion. Although stimulation of BDKRB1 induces weak chemotactic migration of keratinocytes and wound closure in an in vitro scratch-wound assay, the BDKRB1 agonist improved wound closure in a mouse model. BDKRB1 stimulation triggers synthesis and secretion of both metalloproteases, effects that depend on the activity of EGFR and subsequent phosphorylation of ERK1/2 and p38 mitogen-activated protein kinases and PI3K/Akt. In the mouse model, immunoreactivity for both gelatinases was concentrated around wound borders. EGFR transactivation by BDKRB1 agonist involves Src kinases family and ADAM17. In addition to extracellular matrix degradation, metalloproteases 2 and 9 regulate cell migration and differentiation, cell functions that are associated with the role of BDKRB1 in keratinocyte differentiation. Considering that BDKRB1 is up-regulated by inflammation and/or by cytokines that are abundant in the inflammatory milieu, more stable BDKRB1 agonists may be of therapeutic value to modulate wound healing.

  18. Molecular detection and analysis of a novel metalloprotease gene of entomopathogenic Serratia marcescens strains in infected Galleria mellonella.

    PubMed

    Tambong, J T; Xu, R; Sadiku, A; Chen, Q; Badiss, A; Yu, Q

    2014-04-01

    Serratia marcescens strains isolated from entomopathogenic nematodes (Rhabditis sp.) were examined for their pathogenicity and establishment in wax moth (Galleria mellonella) larvae. All the Serratia strains were potently pathogenic to G. mellonella larvae, leading to death within 48 h. The strains were shown to possess a metalloprotease gene encoding for a novel serralysin-like protein. Rapid establishment of the bacteria in infected larvae was confirmed by specific polymerase chain reaction (PCR) detection of a DNA fragment encoding for this protein. Detection of the viable Serratia strains in infected larvae was validated using the SYBR Green reverse transcriptase real-time PCR assay targeting the metalloprotease gene. Nucleotide sequences of the metalloprotease gene obtained in our study showed 72 single nucleotide polymorphisms (SNP) and 3 insertions compared with the metalloprotease gene of S. marcescens E-15. The metalloprotease gene had 60 synonymous and 8 nonsynonymous substitutions relative to the closest GenBank entry, S. marcescens E-15. A comparison of the amino acid composition of the new serralysin-like protein with that of the serralysin protein of S. marcescens E-15 revealed differences at 11 positions and a new aspartic acid residue. Analysis of the effect of protein variation suggests that a new aspartic acid residue resulting from nonsynonymous nucleotide mutations in the protein structure could have the most significant effect on its biological function. The new metalloprotease gene and (or) its product could have applications in plant agricultural biotechnology.

  19. ADAMTS-13 metalloprotease abnormalities in systemic lupus erythematosus: is there a correlation with disease status?

    PubMed

    Klonizakis, P; Tselios, K; Sarantopoulos, A; Gougourellas, I; Rouka, E; Onufriadou, Z; Kapali, P; Kyriakou, D; Boura, P

    2013-04-01

    To clarify the role of ADAMTS-13 in the pathogenesis of thrombotic microangiopathy in systemic lupus erythematosus (SLE) we evaluated ADAMTS-13 profile (metalloprotease antigen levels, anti-ADAMTS-13 autoantibody levels, activity) in distinct patient groups according to disease activity, extent of cumulative tissue damage and history of antiphospholipid syndrome or end-organ damage. Forty-one lupus patients were analysed. ADAMTS-13 metalloprotease antigen levels and anti-ADAMTS-13 autoantibodies were evaluated by ELISA. ADAMTS-13 activity was measured by Fluorescence resonance energy transfer (FRET) technique. ADAMTS-13 metalloprotease antigen levels were significantly decreased in patients with Systemic Lupus International Collaborative Clinics/American College of Rheumatology (SLICC/ACR) >1 (p<0.05). ADAMTS-13 metalloprotease antigen levels also exhibited a significant inverse correlation with anti-dsDNA levels (r= -0.60, p<0.05). Anti-ADAMTS-13 autoantibodies were marginally higher in patients with positive anti-dsDNA (p=0.08). Additionally, patients with positive anti-ADAMTS-13 autoantibodies exhibited the lowest activity levels (p<0.05). To our knowledge ADAMTS-13 profile in SLE has not been studied in regard to composite structured indices. The results of this study suggest that in patients with active SLE or considerable cumulative tissue damage, ADAMTS-13 levels may be decreased and anti-ADAMTS-13 autoantibodies may partially mediate this reduction. Further evaluation of ADAMTS-13 profile may explain its role in the pathogenesis of thrombotic microangiopathy in lupus patients and reveal a potential prognostic marker of microthrombotic manifestations in SLE.

  20. Comparison of the Internal Dynamics of Metalloproteases Provides New Insights on Their Function and Evolution

    PubMed Central

    Carvalho, Henrique F.; Roque, Ana C. A.; Iranzo, Olga; Branco, Ricardo J. F.

    2015-01-01

    Metalloproteases have evolved in a vast number of biological systems, being one of the most diverse types of proteases and presenting a wide range of folds and catalytic metal ions. Given the increasing understanding of protein internal dynamics and its role in enzyme function, we are interested in assessing how the structural heterogeneity of metalloproteases translates into their dynamics. Therefore, the dynamical profile of the clan MA type protein thermolysin, derived from an Elastic Network Model of protein structure, was evaluated against those obtained from a set of experimental structures and molecular dynamics simulation trajectories. A close correspondence was obtained between modes derived from the coarse-grained model and the subspace of functionally-relevant motions observed experimentally, the later being shown to be encoded in the internal dynamics of the protein. This prompted the use of dynamics-based comparison methods that employ such coarse-grained models in a representative set of clan members, allowing for its quantitative description in terms of structural and dynamical variability. Although members show structural similarity, they nonetheless present distinct dynamical profiles, with no apparent correlation between structural and dynamical relatedness. However, previously unnoticed dynamical similarity was found between the relevant members Carboxypeptidase Pfu, Leishmanolysin, and Botulinum Neurotoxin Type A, despite sharing no structural similarity. Inspection of the respective alignments shows that dynamical similarity has a functional basis, namely the need for maintaining proper intermolecular interactions with the respective substrates. These results suggest that distinct selective pressure mechanisms act on metalloproteases at structural and dynamical levels through the course of their evolution. This work shows how new insights on metalloprotease function and evolution can be assessed with comparison schemes that incorporate

  1. A Disintegrin and Metalloprotease 17 in the Cardiovascular and Central Nervous Systems

    PubMed Central

    Xu, Jiaxi; Mukerjee, Snigdha; Silva-Alves, Cristiane R. A.; Carvalho-Galvão, Alynne; Cruz, Josiane C.; Balarini, Camille M.; Braga, Valdir A.; Lazartigues, Eric; França-Silva, Maria S.

    2016-01-01

    ADAM17 is a metalloprotease and disintegrin that lodges in the plasmatic membrane of several cell types and is able to cleave a wide variety of cell surface proteins. It is somatically expressed in mammalian organisms and its proteolytic action influences several physiological and pathological processes. This review focuses on the structure of ADAM17, its signaling in the cardiovascular system and its participation in certain disorders involving the heart, blood vessels, and neural regulation of autonomic and cardiovascular modulation. PMID:27803674

  2. Modulation of the epithelial sodium channel (ENaC) by bacterial metalloproteases and protease inhibitors.

    PubMed

    Butterworth, Michael B; Zhang, Liang; Liu, Xiaoning; Shanks, Robert M; Thibodeau, Patrick H

    2014-01-01

    The serralysin family of metalloproteases is associated with the virulence of multiple gram-negative human pathogens, including Pseudomonas aeruginosa and Serratia marcescens. The serralysin proteases share highly conserved catalytic domains and show evolutionary similarity to the mammalian matrix metalloproteases. Our previous studies demonstrated that alkaline protease (AP) from Pseudomonas aeruginosa is capable of activating the epithelial sodium channel (ENaC), leading to an increase in sodium absorption in airway epithelia. The serralysin proteases are often co-expressed with endogenous, intracellular or periplasmic inhibitors, which putatively protect the bacterium from unwanted or unregulated protease activities. To evaluate the potential use of these small protein inhibitors in regulating the serralysin induced activation of ENaC, proteases from Pseudomonas aeruginosa and Serratia marcescens were purified for characterization along with a high affinity inhibitor from Pseudomonas. Both proteases showed activity against in vitro substrates and could be blocked by near stoichiometric concentrations of the inhibitor. In addition, both proteases were capable of activating ENaC when added to the apical surfaces of multiple epithelial cells with similar slow activation kinetics. The high-affinity periplasmic inhibitor from Pseudomonas effectively blocked this activation. These data suggest that multiple metalloproteases are capable of activating ENaC. Further, the endogenous, periplasmic bacterial inhibitors may be useful for modulating the downstream effects of the serralysin virulence factors under physiological conditions.

  3. Identification and purification of metalloprotease from dry grass pea (Lathyrus sativus L.) seeds.

    PubMed

    Ramakrishna, Vadde; Rajasekhar, Sake; Reddy, Lokireddy Sudarsana

    2010-01-01

    Proteolytic enzymes play a central role in the biochemical mechanism of germination. The present study reported the presence of Zn(2+)-dependent endoproteases in the dry seeds of grass pea (Lathyrus sativus L.) with maximum caseinolytic activity observed at pH 8.0. Studies with class-specific inhibitors (specific for cysteine, serine, aspartate, and metalloproteases) on crude extract identified the inhibitory effect of 1,10-phenanthroline. This inhibitory effect was overcome by addition of Zn(2+), not with Fe, Ca, Cu, Mg, or Co and indicates that the protease is Zn(2+) dependent. This metalloprotease was further characterized by attempting gelatin-PAGE zymography and observed three distinct zones of proteolytic activity with higher mobility. The protease fraction consisted of three isoforms as evidenced by the appearance of three different bands on gelatin-PAGE zymogram. We also purified these proteases to 110-fold by a three-step procedure comprising crude extract from dry seeds, (NH(4))(2)SO(4) fractionation, and casein-alginate affinity chromatography. The molecular mass of isoforms of metalloproteases is 25, 18, and 14 kDa.

  4. Mitochondrial impairment induced by 3-nitropropionic acid is enhanced by endogenous metalloprotease activity inhibition in cultured rat striatal neurons.

    PubMed

    de Oca Balderas, Pavel Montes; Ospina, Gabriel Gutiérrez; Del Ángel, Abel Santamaría

    2013-06-24

    Metalloproteases from the metzincin family mediate molecule processing at the cell membrane termed ectodomain shedding (ES). This mechanism enables the generation of intracellular and extracellular fragments from cell membrane molecules that exert additional functions involved in cell processes including cell death, beyond those of full length molecules. Micotoxin 3-nitropropionic acid (3-NP) induces striatal neuronal degeneration in vivo and in vitro through mitochondrial complex II inhibition. In this study, we hypothesized that metalloproteases regulate mitochondrial activity in cultured rat striatal neurons undergoing degeneration. To test this idea, striatal neuronal cultures characterized by NeuN and GAD-67 expression were treated with 3-NP together with the metalloprotease inhibitor GM6001 and their mitochondrial activity was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Our results showed that metalloprotease inhibition potentiated mitochondrial activity impairment induced by 3-NP whereas the inhibitor alone had no effect. These results indicate that metalloproteases regulate and promote mitochondrial functionality in striatal neurons undergoing degeneration induced by 3-NP. Since NMDA receptor is involved in the excitotoxic neuronal death triggered by 3-NP and is known to undergo ES, we analyzed NMDAR subunit NR1 phenotypic distribution by immunofluorescence. 3-NP and GM6001 induced abnormal perinuclear NR1 accumulation that was not observed with 3-NP or GM6001 alone. This observation suggests that metalloproteases are involved in NR1 cellular reorganization induced by 3-NP, and that their inhibition results in abnormal NR1 distribution. Together results indicate that endogenous metalloproteases are activated during striatal neurodegeneration induced by 3-NP eliciting an adaptative or compensatory response that protects mitochondrial functionality.

  5. Immune cells and mediators involved in the inflammatory responses induced by a P-I metalloprotease and a phospholipase A2 from Bothrops atrox venom.

    PubMed

    Menaldo, Danilo L; Bernardes, Carolina P; Zoccal, Karina F; Jacob-Ferreira, Anna L; Costa, Tássia R; Del Lama, Maria P F M; Naal, Rose M Z G; Frantz, Fabiani G; Faccioli, Lúcia H; Sampaio, Suely V

    2017-05-01

    Bothrops envenomations can promote severe inflammatory responses by inducing edema, pain, leukocyte recruitment and release of chemical mediators by local cells. In the present study, two toxins from Bothrops atrox venom (the P-I metalloprotease Batroxase and the acidic phospholipase A2 BatroxPLA2) were evaluated in relation to their inflammatory effects induced in vivo and in vitro, mainly focusing on the participation of different immune cells and inflammatory mediators. Both toxins mainly promoted acute inflammatory responses with significant recruitment of neutrophils in the early hours (1-4h) after administration into the peritoneal cavity of C57BL/6 mice, and increased infiltration of mononuclear cells especially after 24h. Among the mediators induced by both toxins are IL-6, IL-10 and PGE2, with Batroxase also inducing the release of L-1β, and BatroxPLA2 of LTB4 and CysLTs. These responses pointed to possible involvement of immune cells such as macrophages and mast cells, which were then evaluated in vitro. Mice peritoneal macrophages stimulated with Batroxase produced significant levels of IL-6, IL-1β, PGE2 and LTB4, whereas stimulus with BatroxPLA2 induced increases of IL-6, PGE2 and LTB4. Furthermore, both toxins were able to stimulate degranulation of RBL-2H3 mast cells, but with distinct concentration-dependent effects. Altogether, these results indicated that Batroxase and BatroxPLA2 promoted local and acute inflammatory responses related to macrophages and mast cells and to the production of several mediators. Our findings should contribute for better understanding the different mechanisms of toxicity induced by P-I metalloproteases and phospholipases A2 after snakebite envenomations.

  6. Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

    PubMed Central

    Li, Shangyong; Wang, Linna; Xu, Ximing; Lin, Shengxiang; Wang, Yuejun; Hao, Jianhua; Sun, Mi

    2016-01-01

    Metalloproteases are emerging as useful agents in the treatment of many diseases including arthritis, cancer, cardiovascular diseases, and fibrosis. Studies that could shed light on the metalloprotease pharmaceutical applications require the pure enzyme. Here, we reported the structure-based design and synthesis of the affinity medium for the efficient purification of metalloprotease using the 4-aminophenylboronic acid (4-APBA) as affinity ligand, which was coupled with Sepharose 6B via cyanuric chloride as spacer. The molecular docking analysis showed that the boron atom was interacting with the hydroxyl group of Ser176 residue, whereas the hydroxyl group of the boronic moiety is oriented toward Leu175 and His177 residues. In addition to the covalent bond between the boron atom and hydroxyl group of Ser176, the spacer between boronic acid derivatives and medium beads contributes to the formation of an enzyme-medium complex. With this synthesized medium, we developed and optimized a one-step purification procedure and applied it for the affinity purification of metalloproteases from three commercial enzyme products. The native metalloproteases were purified to high homogeneity with more than 95% purity. The novel purification method developed in this work provides new opportunities for scientific, industrial and pharmaceutical projects. PMID:28036010

  7. Rosiglitazone attenuates the metalloprotease/anti-metalloprotease imbalance in emphysema induced by cigarette smoke: involvement of extracellular signal-regulated kinase and NFκB signaling

    PubMed Central

    Hou, Gang; Yin, Yan; Han, Dan; Wang, Qiu-yue; Kang, Jian

    2015-01-01

    Objective We investigated how rosiglitazone attenuated cigarette smoke (CS)-induced emphysema in a rat model. In particular, we focused on its possible effects on the imbalance between metalloprotease (MMP) and anti-MMP activity, mitogen-activated protein kinase (MAPK) phosphorylation, and nuclear factor kappa-light-chain-enhancer of activated B cell (NFκB) signaling pathway over-activation. Methods A total of 36 Wistar rats were divided into three groups (n=12 each): animals were exposed to CS for 12 weeks in the absence (the CS group) or presence of 30 mg/kg rosiglitazone (the rosiglitazone-CS [RCS] group); a control group was treated with the rosiglitazone vehicle only, without any CS exposure. Histopathology of lung tissue in all groups was evaluated to grade severity of the disease. Expression levels of peroxisome proliferator-activated receptor γ (PPARγ), MMP2, and MMP9 in lung tissue were determined and compared using Western blotting and immunohistochemistry. Activation of MAPKs, NFκB, and the nuclear factor of kappa light polypeptide gene enhancer in B-cell inhibitor, alpha (IκBα) phosphorylation in lung tissue was examined by Western blotting. Results Emphysema-related pathology, based on inter-alveolar wall distance and alveolar density, was less severe in the RCS group than in the CS group. Compared with the CS group, levels of PPARγ were higher in the RCS group, and levels of MMP2 and MMP9 proteins were lower in the RCS rats. Levels of activated MAPKs and NFκB were also lower, while the IκBαphosphorylation was increased in the lung tissue of RCS rats. Conclusion Our findings suggest that oral administration of rosiglitazone attenuates the metalloprotease activity induced by CS, and the underlying mechanism might involve the activation of signaling pathways dependent on MAPKs or NFκB. Our results further suggest that PPARγ contributes to the pathogenesis of emphysema as well as airway inflammation induced by CS. PMID:25897215

  8. Cloning and expression of ADAM-related metalloproteases in equine laminitis.

    PubMed

    Coyne, Michael J; Cousin, Hélène; Loftus, John P; Johnson, Philip J; Belknap, James K; Gradil, Carlos M; Black, Samuel J; Alfandari, Dominique

    2009-06-15

    Equine laminitis is a debilitating disease affecting the digital laminae that suspend the distal phalanx within the hoof. While the clinical progression of the disease has been well documented, the molecular events associated with its pathogenesis remain largely unknown. Using real time quantitative PCR (RT-qPCR), we have investigated the expression of genes coding for proteins containing a Disintegrin and Metalloprotease domain (ADAM), as well as genes encoding the natural inhibitors of these enzymes (tissue inhibitor of metalloprotease; TIMP) in horses with naturally-acquired (acute, chronic and aggravated chronic clinical cases) or experimentally-induced (black walnut extract (BWE) and starch gruel models) laminitis. Changes in expression of these enzymes and regulators may underlie the pathologic remodeling of lamellar tissue in laminitis. Genes encoding ADAMs involved in inflammation (ADAM-10 and ADAM-17), as well as those implicated in arthritis (ADAMTS-1, ADAMTS-4 and ADAMTS-5) were cloned, and the sequences used to generate specific oligonucleotide primers for the RT-qPCR experiments. Our results show that genes encoding ADAM-10 and ADAM-17 were not induced in most laminitic animals, whereas ADAMTS-4 gene expression was strongly upregulated in nearly all horses with experimentally-induced and naturally-acquired laminitis. The expression of matrix metalloproteases (MMP)-9 and ADAMTS-5 was also increased in many of the laminitic horses. In addition, TIMP-2 gene expression was decreased in most laminitic horses, whereas expression of genes encoding other TIMPs, namely TIMP-1 and TIMP-3, was randomly increased or decreased in the various models. We conclude that increased expression of lamellar ADAMTS-4 is a common feature of laminitis consistent with a central role of the gene product in the pathophysiology of the disease.

  9. Metalloproteases Affecting Blood Coagulation, Fibrinolysis and Platelet Aggregation from Snake Venoms: Definition and Nomenclature of Interaction Sites

    PubMed Central

    Kini, R. Manjunatha; Koh, Cho Yeow

    2016-01-01

    Snake venom metalloproteases, in addition to their contribution to the digestion of the prey, affect various physiological functions by cleaving specific proteins. They exhibit their activities through activation of zymogens of coagulation factors, and precursors of integrins or receptors. Based on their structure–function relationships and mechanism of action, we have defined classification and nomenclature of functional sites of proteases. These metalloproteases are useful as research tools and in diagnosis and treatment of various thrombotic and hemostatic conditions. They also contribute to our understanding of molecular details in the activation of specific factors involved in coagulation, platelet aggregation and matrix biology. This review provides a ready reference for metalloproteases that interfere in blood coagulation, fibrinolysis and platelet aggregation. PMID:27690102

  10. Wss1 metalloprotease partners with Cdc48/Doa1 in processing genotoxic SUMO conjugates

    PubMed Central

    Balakirev, Maxim Y; Mullally, James E; Favier, Adrien; Assard, Nicole; Sulpice, Eric; Lindsey, David F; Rulina, Anastasia V; Gidrol, Xavier; Wilkinson, Keith D

    2015-01-01

    Sumoylation during genotoxic stress regulates the composition of DNA repair complexes. The yeast metalloprotease Wss1 clears chromatin-bound sumoylated proteins. Wss1 and its mammalian analog, DVC1/Spartan, belong to minigluzincins family of proteases. Wss1 proteolytic activity is regulated by a cysteine switch mechanism activated by chemical stress and/or DNA binding. Wss1 is required for cell survival following UV irradiation, the smt3-331 mutation and Camptothecin-induced formation of covalent topoisomerase 1 complexes (Top1cc). Wss1 forms a SUMO-specific ternary complex with the AAA ATPase Cdc48 and an adaptor, Doa1. Upon DNA damage Wss1/Cdc48/Doa1 is recruited to sumoylated targets and catalyzes SUMO chain extension through a newly recognized SUMO ligase activity. Activation of Wss1 results in metalloprotease self-cleavage and proteolysis of associated proteins. In cells lacking Tdp1, clearance of topoisomerase covalent complexes becomes SUMO and Wss1-dependent. Upon genotoxic stress, Wss1 is vacuolar, suggesting a link between genotoxic stress and autophagy involving the Doa1 adapter. DOI: http://dx.doi.org/10.7554/eLife.06763.001 PMID:26349035

  11. An oxidant- and organic solvent-resistant alkaline metalloprotease from Streptomyces olivochromogenes.

    PubMed

    Simkhada, Jaya Ram; Cho, Seung Sik; Park, Seong Ju; Mander, Poonam; Choi, Yun Hee; Lee, Hyo Jeong; Yoo, Jin Cheol

    2010-11-01

    Organic solvent- and detergent-resistant proteases are important from an industrial viewpoint. However, they have been less frequently reported and only few of them are from actinomycetes. A metalloprotease from Streptomyces olivochromogenes (SOMP) was purified by ion exchange with Poros HQ and gel filtration with Sepharose CL-6B. Apparent molecular mass of the enzyme was estimated to be 51 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gelatin zymography. The activity was optimum at pH 7.5 and 50 degrees C and stable between pH 7.0 and 10.0. SOMP was stable below 45 degrees C and Ca(2+) increased its thermostability. Ca(2+) enhanced while Co(2+), Cu(2+), Zn(2+), Mn(2+), and Fe(2+) inhibited the activity. Ethylenediaminetetraacetic acid and ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, but not phenylmethylsulfonyl fluoride, aprotinin, and pefabloc SC, significantly suppressed the activity, suggesting that it might be a metalloprotease. Importantly, it is highly resistant against various detergents, organic solvents, and oxidizing agents, and the activity is enhanced by H(2)O(2). The enzyme could be a novel protease based on its origin and peculiar biochemical properties. It may be useful in biotechnological applications especially for organic solvent-based enzymatic synthesis.

  12. Notch modulates VEGF action in endothelial cells by inducing Matrix Metalloprotease activity

    PubMed Central

    2011-01-01

    Background In the vasculature, Notch signaling functions as a downstream effecter of Vascular Endothelial Growth Factor (VEGF) signaling. VEGF regulates sprouting angiogenesis in part by inducing and activating matrix metalloproteases (MMPs). This study sought to determine if VEGF regulation of MMPs was mediated via Notch signaling and to determine how Notch regulation of MMPs influenced endothelial cell morphogenesis. Methods and Results We assessed the relationship between VEGF and Notch signaling in cultured human umbilical vein endothelial cells. Overexpression of VEGF-induced Notch4 and the Notch ligand, Dll4, activated Notch signaling, and altered endothelial cell morphology in a fashion similar to that induced by Notch activation. Expression of a secreted Notch antagonist (Notch1 decoy) suppressed VEGF-mediated activation of endothelial Notch signaling and endothelial morphogenesis. We demonstrate that Notch mediates VEGF-induced matrix metalloprotease activity via induction of MMP9 and MT1-MMP expression and activation of MMP2. Introduction of a MMP inhibitor blocked Notch-mediated endothelial morphogenesis. In mice, analysis of VEGF-induced dermal angiogenesis demonstrated that the Notch1 decoy reduced perivascular MMP9 expression. Conclusions Taken together, our data demonstrate that Notch signaling can act downstream of VEGF signaling to regulate endothelial cell morphogenesis via induction and activation of specific MMPs. In a murine model of VEGF-induced dermal angiogenesis, Notch inhibition led to reduced MMP9 expression. PMID:21349159

  13. Consequences and utility of the zinc-dependent metalloprotease activity of anthrax lethal toxin.

    PubMed

    Bromberg-White, Jennifer; Lee, Chih-Shia; Duesbery, Nicholas

    2010-05-01

    Anthrax is caused by the gram-positive bacterium Bacillus anthracis. The pathogenesis of this disease is dependent on the presence of two binary toxins, edema toxin (EdTx) and lethal toxin (LeTx). LeTx, the major virulence factor contributing to anthrax, contains the effector moiety lethal factor (LF), a zinc-dependent metalloprotease specific for targeting mitogen-activated protein kinase kinases. This review will focus on the protease-specific activity and function of LF, and will include a discussion on the implications and consequences of this activity, both in terms of anthrax disease, and how this activity can be exploited to gain insight into other pathologic conditions.

  14. Microbial biodegradation of proteinaceous tannery solid waste and production of a novel value added product - Metalloprotease.

    PubMed

    Ravindran, Balasubramani; Wong, Jonathan W C; Selvam, Ammaiyappan; Thirunavukarasu, Kathirvel; Sekaran, Ganesan

    2016-10-01

    In this study, animal fleshing (ANFL) was utilized as a substrate for the production of extracellular protease by Clostridium limosum through central composite rotatable design (CCRD) and response surface methodology (RSM). Optimum protease production of 433U/ml was achieved and the purified enzyme was identified as acidic metalloprotease, a monomeric protein. The molecular weight of the enzyme was 71kDa, whose activity was enhanced by bivalent metals such as Zn(2+) and Mg(2+). Scanning electron microscopy (SEM) examination also revealed the hydrolysis/microbial degradation of ANFL through protease activity in the anaerobic fermentation process. Simultaneous hydrolysis of ANFL and production of an enzyme with the potential for different industrial applications provide an attractive methodology for the disposal of tannery solid waste.

  15. Correlated structural kinetics and retarded solvent dynamics at the metalloprotease active site

    SciTech Connect

    Grossman, Moran; Born, Benjamin; Heyden, Matthias; Tworowski, Dmitry; Fields, Gregg B.; Sagi, Irit; Havenith, Martina

    2011-09-18

    Solvent dynamics can play a major role in enzyme activity, but obtaining an accurate, quantitative picture of solvent activity during catalysis is quite challenging. Here, we combine terahertz spectroscopy and X-ray absorption analyses to measure changes in the coupled water-protein motions during peptide hydrolysis by a zinc-dependent human metalloprotease. These changes were tightly correlated with rearrangements at the active site during the formation of productive enzyme-substrate intermediates and were different from those in an enzyme–inhibitor complex. Molecular dynamics simulations showed a steep gradient of fast-to-slow coupled protein-water motions around the protein, active site and substrate. Our results show that water retardation occurs before formation of the functional Michaelis complex. We propose that the observed gradient of coupled protein-water motions may assist enzyme-substrate interactions through water-polarizing mechanisms that are remotely mediated by the catalytic metal ion and the enzyme active site.

  16. Massively parallel enzyme kinetics reveals the substrate recognition landscape of the metalloprotease ADAMTS13.

    PubMed

    Kretz, Colin A; Dai, Manhong; Soylemez, Onuralp; Yee, Andrew; Desch, Karl C; Siemieniak, David; Tomberg, Kärt; Kondrashov, Fyodor A; Meng, Fan; Ginsburg, David

    2015-07-28

    Proteases play important roles in many biologic processes and are key mediators of cancer, inflammation, and thrombosis. However, comprehensive and quantitative techniques to define the substrate specificity profile of proteases are lacking. The metalloprotease ADAMTS13 regulates blood coagulation by cleaving von Willebrand factor (VWF), reducing its procoagulant activity. A mutagenized substrate phage display library based on a 73-amino acid fragment of VWF was constructed, and the ADAMTS13-dependent change in library complexity was evaluated over reaction time points, using high-throughput sequencing. Reaction rate constants (kcat/KM) were calculated for nearly every possible single amino acid substitution within this fragment. This massively parallel enzyme kinetics analysis detailed the specificity of ADAMTS13 and demonstrated the critical importance of the P1-P1' substrate residues while defining exosite binding domains. These data provided empirical evidence for the propensity for epistasis within VWF and showed strong correlation to conservation across orthologs, highlighting evolutionary selective pressures for VWF.

  17. MMP-25 Metalloprotease Regulates Innate Immune Response through NF-κB Signaling.

    PubMed

    Soria-Valles, Clara; Gutiérrez-Fernández, Ana; Osorio, Fernando G; Carrero, Dido; Ferrando, Adolfo A; Colado, Enrique; Fernández-García, M Soledad; Bonzon-Kulichenko, Elena; Vázquez, Jesús; Fueyo, Antonio; López-Otín, Carlos

    2016-07-01

    Matrix metalloproteases (MMPs) regulate innate immunity acting over proinflammatory cytokines, chemokines, and other immune-related proteins. MMP-25 (membrane-type 6-MMP) is a membrane-bound enzyme predominantly expressed in leukocytes whose biological function has remained largely unknown. We have generated Mmp25-deficient mice to elucidate the in vivo function of this protease. These mutant mice are viable and fertile and do not show any spontaneous phenotype. However, Mmp25-null mice exhibit a defective innate immune response characterized by low sensitivity to bacterial LPS, hypergammaglobulinemia, and reduced secretion of proinflammatory molecules. Moreover, these immune defects can be tracked to a defective NF-κB activation observed in Mmp25-deficient leukocytes. Globally, our findings provide new mechanistic insights into innate immunity through the activity of MMP-25, suggesting that this proteinase could be a potential therapeutic target for immune-related diseases.

  18. Growth factor and co-receptor release by structural regulation of substrate metalloprotease accessibility

    PubMed Central

    Parra, Liseth M.; Hartmann, Monika; Schubach, Salome; Ma, Junzhi; Herrlich, Peter; Herrlich, Andreas

    2016-01-01

    Release of cytokines, growth factors and other life-essential molecules from precursors by a-disintegrin-and-metalloproteases (ADAMs) is regulated with high substrate-specificity. We hypothesized that this is achieved by cleavage-regulatory intracellular-domain (ICD)-modifications of the precursors. We show here that cleavage-stimuli-induced specific ICD-modifications cause structural substrate changes that enhance ectodomain sensitivity of neuregulin-1 (NRG1; epidermal-growth-factor) or CD44 (receptor-tyrosine-kinase (RTK) co-receptor) to chymotrypsin/trypsin or soluble ADAM. This inside-out signal transfer required substrate homodimerization and was prevented by cleavage-inhibitory ICD-mutations. In chimeras, regulation could be conferred to a foreign ectodomain, suggesting a common higher-order structure. We predict that substrate-specific protease-accessibility-regulation controls release of numerous ADAM substrates. PMID:27876763

  19. The Metalloprotease Mpl Supports Listeria monocytogenes Dissemination through Resolution of Membrane Protrusions into Vacuoles

    PubMed Central

    Alvarez, Diego E.

    2016-01-01

    Listeria monocytogenes is an intracellular pathogen that disseminates within the intestinal epithelium through acquisition of actin-based motility and formation of plasma membrane protrusions that project into adjacent cells. The resolution of membrane protrusions into vacuoles from which the pathogen escapes results in bacterial spread from cell to cell. This dissemination process relies on the mlp-actA-plcB operon, which encodes ActA, a bacterial nucleation-promoting factor that mediates actin-based motility, and PlcB, a phospholipase that mediates vacuole escape. Here we investigated the role of the metalloprotease Mpl in the dissemination process. In agreement with previous findings showing that Mpl is required for PlcB activation, infection of epithelial cells with the ΔplcB or Δmpl strains resulted in the formation of small infection foci. As expected, the ΔplcB strain displayed a strong defect in vacuole escape. However, the Δmpl strain showed an unexpected defect in the resolution of protrusions into vacuoles, in addition to the expected but mild defect in vacuole escape. The Δmpl strain displayed increased levels of ActA on the bacterial surface in protrusions. We mapped an Mpl-dependent processing site in ActA between amino acid residues 207 to 238. Similar to the Δmpl strain, the ΔactA207–238 strain displayed increased levels of ActA on the bacterial surface in protrusions. Although the ΔactA207–238 strain displayed wild-type actin-based motility, it formed small infection foci and failed to resolve protrusions into vacuoles. We propose that, in addition to its role in PlcB processing and vacuole escape, the metalloprotease Mpl is required for ActA processing and protrusion resolution. PMID:27068088

  20. Inhibition of biosynthesis of metalloprotease of Aeromonas sobria by sodium chloride in the medium.

    PubMed

    Takahashi, Eizo; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Nair, Gopinath Balakrish; Takeda, Yoshifumi; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2011-01-01

    The present authors have previously shown that the serine protease activity of Aeromonas sobria is markedly decreased when A. sobria is cultured in medium containing 3.0% sodium chloride (NaCl, concentration almost equivalent to sea water salinity), and that this occurs because, although the synthesis of ASP is not disturbed by the salt in the medium, the maturation pathway of serine protease of A. sobria (ASP) does not proceed successfully in such a medium. In this study, the effect of salt in the medium on the production of metalloprotease by A. sobria (AMP) was examined. A. sobria produced AMP in the milieu when the bacteria were cultured in medium containing (NaCl) at a concentration of 0.5%. However, AMP was not produced when the bacteria were cultured in salty medium containing 1.5% or more NaCl. To examine how NaCl reduces the production of metalloprotease by A. sobria, the amount of amp mRNA in the cell was measured and it was found that this decreased in proportion to the concentration of NaCl in the medium. The mRNA of amp was not detected in cells cultured in medium containing 1.5% or more NaCl. This means that the transcription of amp is inhibited in salty condition. As described, NaCl in the medium disturbs the maturation pathway of ASP. The mode of action whereby NaCl suppresses AMP activity in A. sobria differs from the mechanism for suppressing ASP activity.

  1. CLCAs - A Family of Metalloproteases of Intriguing Phylogenetic Distribution and with Cases of Substituted Catalytic Sites

    PubMed Central

    Lenart, Anna; Dudkiewicz, Małgorzata; Grynberg, Marcin; Pawłowski, Krzysztof

    2013-01-01

    The zinc-dependent metalloproteases with His-Glu-x-x-His (HExxH) active site motif, zincins, are a broad group of proteins involved in many metabolic and regulatory functions, and found in all forms of life. Human genome contains more than 100 genes encoding proteins with known zincin-like domains. A survey of all proteins containing the HExxH motif shows that approximately 52% of HExxH occurrences fall within known protein structural domains (as defined in the Pfam database). Domain families with majority of members possessing a conserved HExxH motif include, not surprisingly, many known and putative metalloproteases. Furthermore, several HExxH-containing protein domains thus identified can be confidently predicted to be putative peptidases of zincin fold. Thus, we predict zincin-like fold for eight uncharacterised Pfam families. Besides the domains with the HExxH motif strictly conserved, and those with sporadic occurrences, intermediate families are identified that contain some members with a conserved HExxH motif, but also many homologues with substitutions at the conserved positions. Such substitutions can be evolutionarily conserved and non-random, yet functional roles of these inactive zincins are not known. The CLCAs are a novel zincin-like protease family with many cases of substituted active sites. We show that this allegedly metazoan family has a number of bacterial and archaeal members. An extremely patchy phylogenetic distribution of CLCAs in prokaryotes and their conserved protein domain composition strongly suggests an evolutionary scenario of horizontal gene transfer (HGT) from multicellular eukaryotes to bacteria, providing an example of eukaryote-derived xenologues in bacterial genomes. Additionally, in a protein family identified here as closely homologous to CLCA, the CLCA_X (CLCA-like) family, a number of proteins is found in phages and plasmids, supporting the HGT scenario. PMID:23671590

  2. The cell envelope-bound metalloprotease (camelysin) from Bacillus cereus is a possible pathogenic factor.

    PubMed

    Fricke, B; Drössler, K; Willhardt, I; Schierhorn, A; Menge, S; Rücknagel, P

    2001-09-28

    A novel membrane proteinase of the nosocomial important bacteria species Bacillus cereus (synonyms: camelysin, CCMP) was purified up to homogeneity as was shown by mass spectrometry in its amphiphilic form. Camelysin is a neutral metalloprotease with a molecular mass of 19 kDa. Its unique N-terminus Phe-Phe-Ser-Asp-Lys-Glu-Val-Ser-Asn-Asn-Thr-Phe-Ala-Ala-Gly-Thr-Leu-Asp-Leu-Thr-Leu-Asn-Pro-Lys-Thr-Leu-Val-Asp-(Ile-Lys-Asp)- was not detected in the protein data bases during BLAST searches, but in the partially sequenced genome of Bacillus anthracis, coding for an unknown protein. Cleavage sites of the membrane proteinase for the insulin A- and B-chains were determined by mass spectrometry and N-terminal sequencing. Camelysin prefers cleavage sites in front of aliphatic and hydrophilic amino acid residues (-OH, -SO3H, amido group), avoiding bulky aromatic residues. The internally quenched fluorogenic substrates of the matrix metalloproteases 2 and 7 were cleaved with the highest efficiency at the Leu-decrease-Gly or Leu-decrease-Ala bond with the smaller residue in the P1' position. The protein specificity is broad--all various kinds of casein were cleaved as well as acid-soluble collagen, globin and ovalbumin; intact insulin was destroyed only to a low extent. Actin, collagen type I, fibrinogen, fibrin, alpha2-antiplasmin and alpha1-antitrypsin were cleaved. The protease formed SDS-stable complexes with Glu-plasminogen and antithrombin III, visible after SDS electrophoresis by gold staining and Western blot. The CCMP-plasminogen complex caused a partial activation of plasminogen to plasmin. Camelysin interacts with proteins of the blood coagulation cascade and could facilitate the penetration of fibrin clots and of the extracellular matrix during bacterial invasion.

  3. Domain structure and function of matrix metalloprotease 23 (MMP23): role in potassium channel trafficking.

    PubMed

    Galea, Charles A; Nguyen, Hai M; George Chandy, K; Smith, Brian J; Norton, Raymond S

    2014-04-01

    MMP23 is a member of the matrix metalloprotease family of zinc- and calcium-dependent endopeptidases, which are involved in a wide variety of cellular functions. Its catalytic domain displays a high degree of structural homology with those of other metalloproteases, but its atypical domain architecture suggests that it may possess unique functional properties. The N-terminal MMP23 pro-domain contains a type-II transmembrane domain that anchors the protein to the plasma membrane and lacks the cysteine-switch motif that is required to maintain other MMPs in a latent state during passage to the cell surface. Instead of the C-terminal hemopexin domain common to other MMPs, MMP23 contains a small toxin-like domain (TxD) and an immunoglobulin-like cell adhesion molecule (IgCAM) domain. The MMP23 pro-domain can trap Kv1.3 but not closely-related Kv1.2 channels in the endoplasmic reticulum, preventing their passage to the cell surface, while the TxD can bind to the channel pore and block the passage of potassium ions. The MMP23 C-terminal IgCAM domain displays some similarity to Ig-like C2-type domains found in IgCAMs of the immunoglobulin superfamily, which are known to mediate protein-protein and protein-lipid interactions. MMP23 and Kv1.3 are co-expressed in a variety of tissues and together are implicated in diseases including cancer and inflammatory disorders. Further studies are required to elucidate the mechanism of action of this unique member of the MMP family.

  4. Molecular cloning and characterization of Ac-MTP-2, an astacin-like metalloprotease released by adult Ancylostoma caninum.

    PubMed

    Feng, Jianjun; Zhan, Bin; Liu, Yueyuan; Liu, Sen; Williamson, Angela; Goud, Gaddam; Loukas, Alex; Hotez, Peter

    2007-04-01

    Ac-MTP-2 is an astacin-like metalloprotease secreted by adult Ancylostoma caninum hookworms. Ac-mtp-2 cDNA was cloned by immunoscreening a cDNA library with antisera prepared against adult A. caninum excretory/secretory (ES) products. The full-length Ac-mtp-2 contains 850 bp cDNA encoding a 233 amino acid open reading frame (ORF) with 32% amino acid identity to Ce-NSP-4, a pharyngeal cell-derived secreted metalloprotease of the nematode Caenorhabditis elegans. The predicted ORF contained a conserved Met-turn sequence (SXMHY), but only a partial zinc-binding signature sequence (GXXXEHXRXER instead of HEXXHXXGXXHEXXRXDR) found in other astacins. However, by both gelatin gel electrophoresis and azocasein digestion, the recombinant Ac-MTP-2 exhibited proteolytic activity that was inhibited by the zinc chelator 1,10-phenanthroline and Ac-TMP, a putative tissue inhibitor of metalloprotease that was previously shown to be a highly abundant component of adult A. caninum ES products. By RT-PCR, Western blot Ac-MTP-2 was found only expressed in adult hookworms and secreted in the adult ES products. Immunolocalization with antisera shows that Ac-MTP-2 is located to the esophageal glands (confirming its role as a secretory protein), as well as to the parasite uterus. It is hypothesized that Ac-MTP-2 functions in the extracorporeal digestion of the intestinal mucosal plug lodged in the buccal capsule of the adult parasite.

  5. Ikaros and its interacting partner CtBP target the metalloprotease ADAMTS10 to modulate pituitary cell function.

    PubMed

    Shen, Zhongyi; Asa, Sylvia L; Ezzat, Shereen

    2017-01-05

    We have previously described the expression and up-regulation of the C-terminal Binding Protein (CtBP) in response to pituitary hypoxia. This co-repressor interacts with the hematopoietic factor Ikaros to target several components implicated in cellular growth and apoptotic pathways. To identify common transcriptional pituitary targets we performed promoter arrays using Ikaros and CtBP chromatin immunoprecipitated (ChIP) DNA from pituitary AtT20 cells. This approach yielded a finite list of gene targets common to both transcription factors. Of these, the metalloprotease ADAMTS10 emerged as a validated target. We show the ability of Ikaros to bind the ADAMTS10 promoter, influence its transfected activity, and induce endogenous gene expression. ADAMTS10 is expressed in primary pituitary cells and is down-regulated in Ikaros null mice. Further, knockdown of ADAMTS10 in AtT20 cells recapitulates the impact of Ikaros deficiency on POMC/ACTH hormone expression. These results uncover a novel role for the metalloprotease ADAMTS10 in the pituitary. Additionally, they position this metalloprotease as a potential functional integrator of the Ikaros-CtBP chromatin remodeling network.

  6. Two Paralogous Tetraspanins TSP-12 and TSP-14 Function with the ADAM10 Metalloprotease SUP-17 to Promote BMP Signaling in Caenorhabditis elegans

    PubMed Central

    Shi, Herong

    2017-01-01

    The highly conserved bone morphogenetic protein (BMP) signaling pathway regulates many developmental and homeostatic processes. While the core components of the BMP pathway have been well studied, much research is needed for understanding the mechanisms involved in the precise spatiotemporal control of BMP signaling in vivo. Here, we provide evidence that two paralogous and evolutionarily conserved tetraspanins, TSP-12 and TSP-14, function redundantly to promote BMP signaling in C. elegans. We further show that the ADAM10 (a disintegrin and metalloprotease 10) ortholog SUP-17 also functions to promote BMP signaling, and that TSP-12 can bind to and promote the cell surface localization of SUP-17. SUP-17/ADAM10 is known to be involved in the ligand-induced proteolytic processing of the Notch receptor. We have evidence that the function of SUP-17, and of TSP-12/TSP-14 in BMP signaling is independent of their roles in Notch signaling. Furthermore, presenilins, core components of the γ-secretase complex involved in processing Notch, do not appear to play a role in BMP signaling. These studies established a new role of the TSP-12/TSP-14/SUP-17 axis in regulating BMP signaling, in addition to their known function in the Notch signaling pathway. We also provide genetic evidence showing that a known BMP signaling modulator, UNC-40/neogenin/DCC, is one of the substrates of SUP-17/ADAM10 in the BMP signaling pathway. PMID:28068334

  7. SPRTN is a mammalian DNA-binding metalloprotease that resolves DNA-protein crosslinks

    PubMed Central

    Lopez-Mosqueda, Jaime; Maddi, Karthik; Prgomet, Stefan; Kalayil, Sissy; Marinovic-Terzic, Ivana; Terzic, Janos; Dikic, Ivan

    2016-01-01

    Ruijs-Aalfs syndrome is a segmental progeroid syndrome resulting from mutations in the SPRTN gene. Cells derived from patients with SPRTN mutations elicit genomic instability and people afflicted with this syndrome developed hepatocellular carcinoma. Here we describe the molecular mechanism by which SPRTN contributes to genome stability and normal cellular homeostasis. We show that SPRTN is a DNA-dependent mammalian protease required for resolving cytotoxic DNA-protein crosslinks (DPCs)— a function that had only been attributed to the metalloprotease Wss1 in budding yeast. We provide genetic evidence that SPRTN and Wss1 function distinctly in vivo to resolve DPCs. Upon DNA and ubiquitin binding, SPRTN can elicit proteolytic activity; cleaving DPC substrates and itself. SPRTN null cells or cells derived from patients with Ruijs-Aalfs syndrome are impaired in the resolution of covalent DPCs in vivo. Collectively, SPRTN is a mammalian protease required for resolving DNA-protein crosslinks in vivo whose function is compromised in Ruijs-Aalfs syndrome patients. DOI: http://dx.doi.org/10.7554/eLife.21491.001 PMID:27852435

  8. Co-distribution of cysteine cathepsins and matrix metalloproteases in human dentin.

    PubMed

    Scaffa, Polliana Mendes Candia; Breschi, Lorenzo; Mazzoni, Annalisa; Vidal, Cristina de Mattos Pimenta; Curci, Rosa; Apolonio, Fabianni; Gobbi, Pietro; Pashley, David; Tjäderhane, Leo; Tersariol, Ivarne Luis Dos Santos; Nascimento, Fábio Dupart; Carrilho, Marcela Rocha

    2017-02-01

    It has been hypothesized that cysteine cathepsins (CTs) along with matrix metalloproteases (MMPs) may work in conjunction in the proteolysis of mature dentin matrix. The aim of this study was to verify simultaneously the distribution and presence of cathepsins B (CT-B) and K (CT-K) in partially demineralized dentin; and further to evaluate the activity of CTs and MMPs in the same tissue. The distribution of CT-B and CT-K in sound human dentin was assessed by immunohistochemistry. A double-immunolabeling technique was used to identify, at once, the occurrence of those enzymes in dentin. Activities of CTs and MMPs in dentin extracts were evaluated spectrofluorometrically. In addition, in situ gelatinolytic activity of dentin was assayed by zymography. The results revealed the distribution of CT-B and CT-K along the dentin organic matrix and also indicated co-occurrence of MMPs and CTs in that tissue. The enzyme kinetics studies showed proteolytic activity in dentin extracts for both classes of proteases. Furthermore, it was observed that, at least for sound human dentin matrices, the activity of MMPs seems to be predominant over the CTs one.

  9. Cloning, sequencing and expression of the gene encoding the extracellular metalloprotease of Aeromonas caviae.

    PubMed

    Kawakami, K; Toma, C; Honma, Y

    2000-01-01

    A gene (apk) encoding the extracellular protease of Aeromonas caviae Ae6 has been cloned and sequenced. For cloning the gene, the DNA genomic library was screened using skim milk LB agar. One clone harboring plasmid pKK3 was selected for sequencing. Nucleotide sequencing of the 3.5 kb region of pKK3 revealed a single open reading frame (ORF) of 1,785 bp encoding 595 amino acids. The deduced polypeptide contained a putative 16-amino acid signal peptide followed by a large propeptide. The N-terminal amino acid sequence of purified recombinant protein (APK) was consistent with the DNA sequence. This result suggested a mature protein of 412 amino acids with a molecular mass of 44 kDa. However, the molecular mass of purified recombinant APK revealed 34 kDa by SDS-PAGE, suggesting that further processing at the C-terminal region took place. The 2 motifs of zinc binding sites deduced are highly conserved in the APK as well as in other zinc metalloproteases including Vibrio proteolyticus neutral protease, Emp V from Vibrio vulnificus, HA/P from Vibrio cholerae, and Pseudomonas aeruginosa elastase. Proteolytic activity was inhibited by EDTA, Zincov, 1,10-phenanthroline and tetraethylenepentamine while unaffected by the other inhibitors tested. The protease showed maximum activity at pH 7.0 and was inactivated by heating at 80 C for 15 min. These results together suggest that APK belongs to the thermolysin family of metalloendopeptidases.

  10. The Metalloprotease Meprin β Is an Alternative β-Secretase of APP

    PubMed Central

    Becker-Pauly, Christoph; Pietrzik, Claus U.

    2017-01-01

    The membrane bound metalloprotease meprin β is important for collagen fibril assembly in connective tissue formation and for the detachment of the intestinal mucus layer for proper barrier function. Recent proteomic studies revealed dozens of putative new substrates of meprin β, including the amyloid precursor protein (APP). It was shown that APP is cleaved by meprin β in distinct ways, either at the β-secretase site resulting in increased levels of Aβ peptides, or at the N-terminus releasing 11 kDa, and 20 kDa peptide fragments. The latter event was discussed to be rather neuroprotective, whereas the ectodomain shedding of APP by meprin β reminiscent to BACE-1 is in line with the amyloid hypothesis of Alzheimer's disease, promoting neurodegeneration. The N-terminal 11 kDa and 20 kDa peptide fragments represent physiological cleavage products, since they are found in human brains under different diseased or non-diseased states, whereas these fragments are completely missing in brains of meprin β knock-out animals. Meprin β is not only a sheddase of adhesion molecules, such as APP, but was additionally demonstrated to cleave within the prodomain of ADAM10. Activated ADAM10, the α-secretase of APP, is then able to shed meprin β from the cell surface thereby abolishing the β-secretase activity. All together meprin β seems to be a novel player in APP processing events, even influencing other enzymes involved in APP cleavage. PMID:28105004

  11. Squaric Acid-Based Peptidic Inhibitors of Matrix Metalloprotease-1 (MMP-1)

    PubMed Central

    Onaran, M. Burak; Comeau, Anthony B.; Seto, Christopher T.

    2008-01-01

    A series of squaric acid-peptide conjugates were synthesized and evaluated as inhibitors of MMP-1. The cyclobut-3-enedione core was substituted at the 3-position with several functional groups, such as -N(alkyl)OH, -NHOH and –OH, that are designed to bind to the zinc atom in the active site of the metalloprotease. The 4-position of the cyclobut-3-enedione was derivatized with mono- or dipeptides that are designed to bind in the S1′ and S2′ subsites of the enzyme, and position the metal chelating group appropriately in the active site for binding to zinc. Positional scanning revealed that -N(Me)OH provided the highest level of inhibition among the chelating groups that were tested, and Leu-Tle-NHMe was the preferred amino acid sequence. A combination of these groups yielded an inhibitor with an IC50 value of 95 μM. For one inhibitor, conversion of one of the carbonyl groups on the cyclobut-3-enedione core to a thiocarbonyl group resulted in a 18-fold increase in potency, and yielded a compound with an IC50 value of 15 μM. PMID:16356002

  12. Metabolism of AGEs – Bacterial AGEs Are Degraded by Metallo-Proteases

    PubMed Central

    Cohen-Or, Ifat; Katz, Chen; Ron, Eliora Z.

    2013-01-01

    Advanced Glycation End Products (AGEs) are the final products of non-enzymatic protein glycation that results in loss of protein structure and function. We have previously shown that in E. coli AGEs are continually formed as high-molecular weight protein complexes. Moreover, we showed that AGEs are removed from the cells by an active, ATP-dependent secretion and that these secreted molecules have low molecular weight. Taken together, these results indicate that E. coli contains a fraction of low molecular weight AGEs, in addition to the high-molecular weight AGEs. Here we show that the low-molecular weight AGEs originate from high-molecular weight AGEs by proteolytic degradation. Results of in-vitro and in vivo experiments indicated that this degradation is carried out not by the major ATP-dependent proteases that are responsible for the main part of bacterial protein quality control but by an alternative metal-dependent proteolysis. This proteolytic reaction is essential for the further secretion of AGEs from the cells. As the biochemical reactions involving AGEs are not yet understood, the implication of a metalloprotease in breakdown of high molecular weight AGEs and their secretion constitutes an important step in the understanding of AGEs metabolism. PMID:24130678

  13. Hyaluronic acid alkyl derivative: A novel inhibitor of metalloproteases and hyaluronidases.

    PubMed

    Pavan, Mauro; Galesso, Devis; Secchieri, Cynthia; Guarise, Cristian

    2016-03-01

    Extracellular matrix (ECM) degradation, one of the main features of osteoarthritis, is driven by at least two major classes of enzymes: matrix metalloproteases (MMPs) and hyaluronidases. Among certain glycosaminoglycans, including natural and chemically cross-linked HAs, which are currently used as viscosupplements, the hyaluronic acid (HA) alkyl-amides (Hyadd) were here selected as the strongest MMP and hyaluronidase inhibitors. We used C. histolyticum collagenase (ChC) and bovine testicular hyaluronidase (BTH) as representative models of human MMPs and hyaluronidases, respectively. The role of the alkyl moiety was investigated using HA derivatives with varying alkyl lengths and degrees of derivatization. The selected compound was then screened against 10 different human MMPs in vitro, and the results were validated ex vivo in human synovial fluid. Hyadd-C16, identified as a lead compound, showed the highest inhibition potency against MMP13 and MMP8. The in vitro results were confirmed by the inhibition of human MMP13 (Ki=106.1 μM) and hyaluronidase-2 in the synovial fluid of patients with osteoarthritis. This study demonstrates the unique properties of Hyadd-C16, including its remarkable enzymatic inhibitory activity, which is conferred by the hydrophobic chain, and its high biocompatibility and water solubility of the HA backbone.

  14. Targeting a Single Function of the Multifunctional Matrix Metalloprotease MT1-MMP

    PubMed Central

    Ingvarsen, Signe; Porse, Astrid; Erpicum, Charlotte; Maertens, Ludovic; Jürgensen, Henrik J.; Madsen, Daniel H.; Melander, Maria C.; Gårdsvoll, Henrik; Høyer-Hansen, Gunilla; Noel, Agnès; Holmbeck, Kenn; Engelholm, Lars H.; Behrendt, Niels

    2013-01-01

    The group of matrix metalloproteases (MMPs) is responsible for multiple processes of extracellular matrix remodeling in the healthy body but also for matrix and tissue destruction during cancer invasion and metastasis. The understanding of the contributions from each individual MMP, both in healthy and pathological events, has been complicated by the lack of specific inhibitors and the fact that some of the potent MMPs are multifunctional enzymes. These factors have also hampered the setup of therapeutic strategies targeting MMP activity. A tempting target is the membrane-associated MT1-MMP, which has well-documented importance in matrix degradation but which takes part in more than one pathway in this regard. In this report, we describe the selective targeting of a single function of this enzyme by means of a specific monoclonal antibody against MT1-MMP, raised in an MT1-MMP knock-out mouse. The antibody blocks the enzyme ability to activate proMMP-2 without interfering with the collagenolytic function or the general proteolytic activity of MT1-MMP. Using this antibody, we have shown that the MT1-MMP-catalyzed activation of proMMP-2 is involved in the outgrowth of cultured lymphatic endothelial cells in a collagen matrix in vitro, as well as in lymphatic vessel sprouting assayed ex vivo. This is the first example of the complete inactivation of a single function of a multifunctional MMP and the use of this strategy to pursue its role. PMID:23413031

  15. Targeting a single function of the multifunctional matrix metalloprotease MT1-MMP: impact on lymphangiogenesis.

    PubMed

    Ingvarsen, Signe; Porse, Astrid; Erpicum, Charlotte; Maertens, Ludovic; Jürgensen, Henrik J; Madsen, Daniel H; Melander, Maria C; Gårdsvoll, Henrik; Høyer-Hansen, Gunilla; Noel, Agnès; Holmbeck, Kenn; Engelholm, Lars H; Behrendt, Niels

    2013-04-12

    The group of matrix metalloproteases (MMPs) is responsible for multiple processes of extracellular matrix remodeling in the healthy body but also for matrix and tissue destruction during cancer invasion and metastasis. The understanding of the contributions from each individual MMP, both in healthy and pathological events, has been complicated by the lack of specific inhibitors and the fact that some of the potent MMPs are multifunctional enzymes. These factors have also hampered the setup of therapeutic strategies targeting MMP activity. A tempting target is the membrane-associated MT1-MMP, which has well-documented importance in matrix degradation but which takes part in more than one pathway in this regard. In this report, we describe the selective targeting of a single function of this enzyme by means of a specific monoclonal antibody against MT1-MMP, raised in an MT1-MMP knock-out mouse. The antibody blocks the enzyme ability to activate proMMP-2 without interfering with the collagenolytic function or the general proteolytic activity of MT1-MMP. Using this antibody, we have shown that the MT1-MMP-catalyzed activation of proMMP-2 is involved in the outgrowth of cultured lymphatic endothelial cells in a collagen matrix in vitro, as well as in lymphatic vessel sprouting assayed ex vivo. This is the first example of the complete inactivation of a single function of a multifunctional MMP and the use of this strategy to pursue its role.

  16. Detection of Intracellular ADAMTS13, a Secreted Zinc-metalloprotease, via Flow Cytometry

    PubMed Central

    S., Geetha; Allen, Courtni E.; Hunt, Ryan; Plum, Elizabeth; Garfield, Susan; Friedman, Scott L.; Soejima, Kenji; Sauna, Zuben E.; Kimchi-Sarfaty, Chava

    2009-01-01

    Background ADAMTS13 is a secreted metalloprotease that cleaves von Willebrand Factor multimers and maintains proper homeostasis. A severe deficiency in ADAMTS13 triggers a disorder known as thrombotic thrombocytopenic purpura (TTP). At present, ADAMTS13 expression levels are determined by immunoblotting. Methods We established a flow cytometry methodology to detect intracellular ADAMTS13 in liver and kidney cells using a polyclonal antibody, BL154G, and several monoclonal antibodies previously used to detect ADAMTS13 by immunoblotting. Results were validated using confocal microscopy, immunoblotting and an activity assay (FRETS-VWF73). Results We show that labeling ADAMTS13 with specific antibodies and detection by flow cytometry yields results that are comparable to previously established methods for ADAMTS13 detection. Specifically, we compared the endogenous expression levels of ADAMTS13 in various liver cell lines using flow cytometry and obtained results that parallel immunoblot analysis. Knock-down of ADAMTS13 expression via targeted siRNA resulted in significantly reduced median signal, displaying the sensitivity of this detection method. A further analysis of reliability and specificity was achieved through plasmid DNA and transfection reagent dose response studies. Conclusions The flow cytometry method described here is useful in determining the expression of both endogenous and recombinant forms of intracellular ADAMTS13. Flow cytometry is a convenient, efficient and cost effective way to measure the expression levels of ADAMTS13. PMID:19526483

  17. A novel matrix metalloprotease-like enzyme (karilysin) of the periodontal pathogen Tannerella forsythia ATCC 43037.

    PubMed

    Karim, Abdulkarim Y; Kulczycka, Magdalena; Kantyka, Tomasz; Dubin, Grzegorz; Jabaiah, Abeer; Daugherty, Patrick S; Thogersen, Ida B; Enghild, Jan J; Nguyen, Ky-Anh; Potempa, Jan

    2010-01-01

    Proteases of Tannerella forsythia, a pathogen associated with periodontal disease, are implicated as virulence factors. Here, we characterized a matrix metalloprotease (MMP)-like enzyme of T. forsythia referred to as karilysin. Full-length (without a signal peptide) recombinant karilysin (49.9 kDa) processed itself into the mature 18-kDa enzyme through sequential autoproteolytic cleavage at both N- and C-terminal profragments. The first cleavage at the Asn14-Tyr15 peptide bond generated the fully active enzyme (47.9 kDa) and subsequent truncations at the C-terminus did not affect proteolytic activity. Mutation of Tyr15 to Ala generated a prokarilysin variant that processed itself into the final 18-kDa form with greatly reduced kinetics. Inactive prokarilysin with the mutated catalytic Glu residue (E136A) was processed by active karilysin at the same sites as the active enzymes. Karilysin proteolytic activity and autoprocessing were inhibited by 1,10-phenanthroline and EDTA. Calcium ions were found to be important for both the activity and thermal stability of karilysin. Using CLiPS technology, the specificity of karilysin was found to be similar to that of MMPs with preference for Leu/Tyr/Met at P1' and Pro/Ala at P3. This specificity and the ability to degrade elastin, fibrinogen and fibronectin may contribute to the pathogenicity of periodontitis.

  18. Hetero- and autoprocessing of the extracellular metalloprotease (Mpr) in Bacillus subtilis.

    PubMed

    Park, Chi Hye; Lee, Sang Jun; Lee, Sung Gu; Lee, Weon Sup; Byun, Si Myung

    2004-10-01

    Most proteases are synthesized as inactive precursors which are processed by proteolytic cleavage into a mature active form, allowing regulation of their proteolytic activity. The activation of the glutamic-acid-specific extracellular metalloprotease (Mpr) of Bacillus subtilis has been examined. Analysis of Mpr processing in defined protease-deficient mutants by activity assay and Western blotting revealed that the extracellular protease Bpr is required for Mpr processing. pro-Mpr remained a precursor form in bpr-deficient strains, and glutamic-acid-specific proteolytic activity conferred by Mpr was not activated in bpr-deficient strains. Further, purified pro-Mpr was processed to an active form by purified Bpr protease in vitro. We conclude that Mpr is activated by Bpr in vivo, and that heteroprocessing, rather than autoprocessing, is the major mechanism of Mpr processing in vivo. Exchange of glutamic acid for serine in the cleavage site of Mpr (S93E) allowed processing of Mpr into its mature form, regardless of the presence of other extracellular proteases, including Bpr. Thus, a single amino acid change is sufficient to convert the Mpr processing mechanism from heteroprocessing to autoprocessing.

  19. A disintegrin and metalloprotease 10 (ADAM10) is a central regulator of murine liver tissue homeostasis

    PubMed Central

    Köhn-Gaone, Julia; Chalupsky, Karel; Lüllmann-Rauch, Renate; Barikbin, Roja; Bergmann, Juri; Wöhner, Birte; Zbodakova, Olga; Leuschner, Ivo; Martin, Gregor; Tiegs, Gisa; Rose-John, Stefan; Sedlacek, Radislav; Tirnitz-Parker, Janina E.E.; Saftig, Paul; Schmidt-Arras, Dirk

    2016-01-01

    A Disintegrin And Metalloprotease (ADAM) 10 exerts essential roles during organ development and tissue integrity in different organs, mainly through activation of the Notch pathway. However, only little is known about its implication in liver tissue physiology. Here we show that in contrast to its role in other tissues, ADAM10 is dispensable for the Notch2-dependent biliary tree formation. However, we demonstrate that expression of bile acid transporters is dependent on ADAM10. Consequently, mice deficient for Adam10 in hepatocytes, cholangiocytes and liver progenitor cells develop spontaneous hepatocyte necrosis and concomitant liver fibrosis. We furthermore observed a strongly augmented ductular reaction in 15-week old ADAM10Δhep/Δch mice and demonstrate that c-Met dependent liver progenitor cell activation is enhanced. Additionally, liver progenitor cells are primed to hepatocyte differentiation in the absence of ADAM10. These findings show that ADAM10 is a novel central node controlling liver tissue homeostasis. Highlights: Loss of ADAM10 in murine liver results in hepatocyte necrosis and concomitant liver fibrosis. ADAM10 directly regulates expression of bile acid transporters but is dispensable for Notch2-dependent formation of the biliary system. Activation of liver progenitor cells is enhanced through increased c-Met signalling, in the absence of ADAM10. Differentiation of liver progenitor cells to hepatocytes is augmented in the absence of ADAM10. PMID:26942887

  20. The Metalloprotease Meprin β Is an Alternative β-Secretase of APP.

    PubMed

    Becker-Pauly, Christoph; Pietrzik, Claus U

    2016-01-01

    The membrane bound metalloprotease meprin β is important for collagen fibril assembly in connective tissue formation and for the detachment of the intestinal mucus layer for proper barrier function. Recent proteomic studies revealed dozens of putative new substrates of meprin β, including the amyloid precursor protein (APP). It was shown that APP is cleaved by meprin β in distinct ways, either at the β-secretase site resulting in increased levels of Aβ peptides, or at the N-terminus releasing 11 kDa, and 20 kDa peptide fragments. The latter event was discussed to be rather neuroprotective, whereas the ectodomain shedding of APP by meprin β reminiscent to BACE-1 is in line with the amyloid hypothesis of Alzheimer's disease, promoting neurodegeneration. The N-terminal 11 kDa and 20 kDa peptide fragments represent physiological cleavage products, since they are found in human brains under different diseased or non-diseased states, whereas these fragments are completely missing in brains of meprin β knock-out animals. Meprin β is not only a sheddase of adhesion molecules, such as APP, but was additionally demonstrated to cleave within the prodomain of ADAM10. Activated ADAM10, the α-secretase of APP, is then able to shed meprin β from the cell surface thereby abolishing the β-secretase activity. All together meprin β seems to be a novel player in APP processing events, even influencing other enzymes involved in APP cleavage.

  1. Reprolysin metalloproteases from Ixodes persulcatus, Rhipicephalus sanguineus and Rhipicephalus microplus ticks.

    PubMed

    Ali, Abid; Tirloni, Lucas; Isezaki, Masayoshi; Seixas, Adriana; Konnai, Satoru; Ohashi, Kazuhiko; da Silva Vaz Junior, Itabajara; Termignoni, Carlos

    2014-08-01

    Metalloproteases (MPs) have been considered essential for blood feeding and other physiological functions in several hematophagous animals, including ticks. We report the characterization of MP sequences of three important ticks from Asia, Africa and America: Ixodes persulcatus (Ip-MPs), Rhipicephalus sanguineus (Rs-MPs) and R. microplus (BrRm-MPs). Amino acid sequence identity between R. microplus and R. sanguineus MPs ranged from 76 to 100 %, and identities among I. persulcatus, I. ricinus and I. scapularis MP sequences ranged from 88 to 97 %. This high sequence identity and typical functional motifs show that all sequences are MPs. The presence of a zinc binding site, a Met-turn and cysteine rich domain at the C-terminal region indicates that these proteins belong to the reproplysin family of MPs. Differences in amino acid sequences of BrRm-MP1, BrRm-MP2, BrRm-MP4 and BrRm-MP5 (from Porto Alegre strain ticks) were 6, 2, 7 and 5 %, respectively, when compared with sequences deposited in GenBank for the same genes from other R. microplus isolates. Analyses of MPs predicted that they have various highly antigenic regions. Semi-quantitative RT-PCR analysis revealed the presence of transcripts in salivary glands of partially and fully fed female ticks. None of these transcripts were observed in males (except BrRm-MP4) and eggs. These enzymes may be functional components required during tick feeding to manipulate host defenses and support tick hematophagy.

  2. Particulate matter (PM₁₀) induces metalloprotease activity and invasion in airway epithelial cells.

    PubMed

    Morales-Bárcenas, Rocío; Chirino, Yolanda I; Sánchez-Pérez, Yesennia; Osornio-Vargas, Álvaro Román; Melendez-Zajgla, Jorge; Rosas, Irma; García-Cuellar, Claudia María

    2015-09-17

    Airborne particulate matter with an aerodynamic diameter ≤ 10 μm (PM10) is a risk factor for the development of lung diseases and cancer. The aim of this work was to identify alterations in airway epithelial (A549) cells induced by PM10 that could explain how subtoxic exposure (10 μg/cm(2)) promotes a more aggressive in vitro phenotype. Our results showed that cells exposed to PM10 from an industrial zone (IZ) and an urban commercial zone (CZ) induced an increase in protease activity and invasiveness; however, the cell mechanism is different, as only PM10 from CZ up-regulated the activity of metalloproteases MMP-2 and MMP-9 and disrupted E-cadherin/β-catenin expression after 48 h of exposure. These in vitro findings are relevant in terms of the mechanism action of PM10 in lung epithelial cells, which could be helpful in understanding the pathogenesis of some human illness associated with highly polluted cities.

  3. Meprin Metalloproteases Generate Biologically Active Soluble Interleukin-6 Receptor to Induce Trans-Signaling

    PubMed Central

    Arnold, Philipp; Boll, Inga; Rothaug, Michelle; Schumacher, Neele; Schmidt, Frederike; Wichert, Rielana; Schneppenheim, Janna; Lokau, Juliane; Pickhinke, Ute; Koudelka, Tomas; Tholey, Andreas; Rabe, Björn; Scheller, Jürgen; Lucius, Ralph; Garbers, Christoph; Rose-John, Stefan; Becker-Pauly, Christoph

    2017-01-01

    Soluble Interleukin-6 receptor (sIL-6R) mediated trans-signaling is an important pro-inflammatory stimulus associated with pathological conditions, such as arthritis, neurodegeneration and inflammatory bowel disease. The sIL-6R is generated proteolytically from its membrane bound form and A Disintegrin And Metalloprotease (ADAM) 10 and 17 were shown to perform ectodomain shedding of the receptor in vitro and in vivo. However, under certain conditions not all sIL-6R could be assigned to ADAM10/17 activity. Here, we demonstrate that the IL-6R is a shedding substrate of soluble meprin α and membrane bound meprin β, resulting in bioactive sIL-6R that is capable of inducing IL-6 trans-signaling. We determined cleavage within the N-terminal part of the IL-6R stalk region, distinct from the cleavage site reported for ADAM10/17. Interestingly, meprin β can be shed from the cell surface by ADAM10/17 and the observation that soluble meprin β is not capable of shedding the IL-6R suggests a regulatory mechanism towards trans-signaling. Additionally, we observed a significant negative correlation of meprin β expression and IL-6R levels on human granulocytes, providing evidence for in vivo function of this proteolytic interaction. PMID:28276471

  4. ORMDL3 is an inducible lung epithelial gene regulating metalloproteases, chemokines, OAS, and ATF6.

    PubMed

    Miller, Marina; Tam, Arvin B; Cho, Jae Youn; Doherty, Taylor A; Pham, Alexa; Khorram, Naseem; Rosenthal, Peter; Mueller, James L; Hoffman, Hal M; Suzukawa, Maho; Niwa, Maho; Broide, David H

    2012-10-09

    Orosomucoid like 3 (ORMDL3) has been strongly linked with asthma in genetic association studies, but its function in asthma is unknown. We demonstrate that in mice ORMDL3 is an allergen and cytokine (IL-4 or IL-13) inducible endoplasmic reticulum (ER) gene expressed predominantly in airway epithelial cells. Allergen challenge induces a 127-fold increase in ORMDL3 mRNA in bronchial epithelium in WT mice, with lesser 15-fold increases in ORMDL-2 and no changes in ORMDL-1. Studies of STAT-6-deficient mice demonstrated that ORMDL3 mRNA induction highly depends on STAT-6. Transfection of ORMDL3 in human bronchial epithelial cells in vitro induced expression of metalloproteases (MMP-9, ADAM-8), CC chemokines (CCL-20), CXC chemokines (IL-8, CXCL-10, CXCL-11), oligoadenylate synthetases (OAS) genes, and selectively activated activating transcription factor 6 (ATF6), an unfolded protein response (UPR) pathway transcription factor. siRNA knockdown of ATF-6α in lung epithelial cells inhibited expression of SERCA2b, which has been implicated in airway remodeling in asthma. In addition, transfection of ORMDL3 in lung epithelial cells activated ATF6α and induced SERCA2b. These studies provide evidence of the inducible nature of ORMDL3 ER expression in particular in bronchial epithelial cells and suggest an ER UPR pathway through which ORMDL3 may be linked to asthma.

  5. Collagenolytic activity related to metalloproteases (and serine proteases) in the fish parasite Hysterothylacium aduncum (Nematoda: Anisakidae).

    PubMed

    Malagón, David; Adroher, Francisco Javier; Díaz-López, Manuel; Benítez, Rocío

    2010-06-11

    Proteases play a vital role in both the life cycle of parasites and the parasite-host relationship and are considered important virulence factors. In the present study, the presence of proteases with collagenolytic activity was investigated in the fish nematode Hysterothylacium aduncum during in vitro development. Collagenolytic activity was found in all studied developmental stages of the nematode (third [L3] and fourth [L4] larval stages and adults). In L3, the activity was maximum at pH 6.5 and, in the other stages, at 7.0. Pepsin is known to favour in vitro development of the worm, but, in this study, collagenolytic activity was shown to be significantly greater when no pepsin was added to the culture medium (at pH 6.5, p = 0.011). At pH 7.0, most activity was observed in the immature adult, after the final moult, suggesting that the collagenolytic activity may be involved in remodelling of the cuticle and in sexual maturity. On the other hand, at pH 6.5, activity may be related to tissue migration by L3 within the host. Using specific inhibitors, it was demonstrated that most of the collagenolytic activity detected in all the developmental stages was due to metalloproteases (40 to 100%), although serine proteases were also detected in L4 and adults (10 to 30%).

  6. The metalloprotease ADAM12 regulates the effector function of human Th17 cells.

    PubMed

    Zhou, Angela X; El Hed, Aimee; Mercer, Frances; Kozhaya, Lina; Unutmaz, Derya

    2013-01-01

    A key modulator of immune homeostasis, TGFβ has an important role in the differentiation of regulatory T cells (Tregs) and IL-17-secreting T cells (Th17). How TGFβ regulates these functionally opposing T cell subsets is not well understood. We determined that an ADAM family metalloprotease called ADAM12 is specifically and highly expressed in both Tregs and CCR6+ Th17 cells. ADAM12 is induced in vitro upon differentiation of naïve T cells to Th17 cells or IL-17-secreting Tregs. Remarkably, silencing ADAM12 expression in CCR6+ memory T cells enhances the production of Th17 cytokines, similar to suppressing TGFβ signaling. Further, ADAM12 knockdown in naïve human T cells polarized towards Th17/Treg cells, or ectopically expressing RORC, greatly enhances IL-17-secreting cell differentiation, more potently then inhibiting TGFβ signals. Together, our findings reveal a novel regulatory role for ADAM12 in Th17 cell differentiation or function and may have implications in regulating their aberrant responses during immune pathologies.

  7. Metalloprotease-disintegrin ADAM12 expression is regulated by Notch signaling via microRNA-29.

    PubMed

    Li, Hui; Solomon, Emilia; Duhachek Muggy, Sara; Sun, Danqiong; Zolkiewska, Anna

    2011-06-17

    Metalloprotease-disintegrin ADAM12 is overexpressed and frequently mutated in breast cancer. We report here that ADAM12 expression in cultured mammalian cells is up-regulated by Notch signals. Expression of a constitutively active form of Notch1 in murine fibroblasts, myoblasts, or mammary epithelial cells or activation of the endogenous Notch signaling by co-culture with ligand-expressing cells increases ADAM12 protein and mRNA levels. Up-regulation of ADAM12 expression by Notch requires new transcription, is activated in a CSL-dependent manner, and is abolished upon inhibition of IκB kinase. Expression of a constitutively active Notch1 in NIH3T3 cells increases the stability of Adam12 mRNA. We further show that the microRNA-29 family, which has a predicted conserved site in the 3'-untranslated region of mouse Adam12, plays a critical role in mediating the stimulatory effect of Notch on ADAM12 expression. In human cells, Notch up-regulates the expression of the long form, but not the short form, of ADAM12 containing a divergent 3'-untranslated mRNA region. These studies uncover a novel paradigm in Notch signaling and establish Adam12 as a Notch-related gene.

  8. Cell-surface metalloprotease ADAM12 is internalized by a clathrin- and Grb2-dependent mechanism.

    PubMed

    Stautz, Dorte; Leyme, Anthony; Grandal, Michael Vibo; Albrechtsen, Reidar; van Deurs, Bo; Wewer, Ulla; Kveiborg, Marie

    2012-11-01

    ADAM12 (A Disintegrin And Metalloprotease 12), a member of the ADAMs family of transmembrane proteins, is involved in ectodomain shedding, cell-adhesion and signaling, with important implications in cancer. Therefore, mechanisms that regulate the levels and activity of ADAM12 at the cell-surface are possibly crucial in these contexts. We here investigated internalization and subsequent recycling or degradation of ADAM12 as a potentially important regulatory mechanism. Our results show that ADAM12 is constitutively internalized primarily via the clathrin-dependent pathway and is subsequently detected in both early and recycling endosomes. The protease activity of ADAM12 does not influence this internalization mechanism. Analysis of essential elements for internalization established that proline-rich regions in the cytoplasmic domain of ADAM12, previously shown to interact with Src-homology 3 domains, were necessary for proper internalization. These sites in the ADAM12 cytoplasmic domain interacted with the adaptor protein growth factor receptor-bound protein 2 (Grb2) and knockdown of Grb2 markedly reduced ADAM12 internalization. These studies establish that internalization is indeed a mechanism that regulates ADAM cell surface levels and show that ADAM12 internalization involves the clathrin-dependent pathway and Grb2.

  9. Plasmatic ADAMTS-13 metalloprotease and von Willebrand factor in children with cyanotic congenital heart disease

    PubMed Central

    Soares, R.P.S.; Bydlowski, S.P.; Nascimento, N.M.; Thomaz, A.M.; Bastos, E.N.M.; Lopes, A.A.

    2013-01-01

    Changes in plasma von Willebrand factor concentration (VWF:Ag) and ADAMTS-13 activity (the metalloprotease that cleaves VWF physiologically) have been reported in several cardiovascular disorders with prognostic implications. We therefore determined the level of these proteins in the plasma of children with cyanotic congenital heart disease (CCHD) undergoing surgical treatment. Forty-eight children were enrolled (age 0.83 to 7.58 years). Measurements were performed at baseline and 48 h after surgery. ELISA, collagen-binding assays and Western blotting were used to estimate antigenic and biological activities, and proteolysis of VWF multimers. Preoperatively, VWF:Ag and ADAMTS-13 activity were decreased (65 and 71% of normal levels considered as 113 (105-129) U/dL and 91 ± 24% respectively, P < 0.003) and correlated (r = 0.39, P = 0.0064). High molecular weight VWF multimers were not related, suggesting an interaction of VWF with cell membranes, followed by proteolytic cleavage. A low preoperative ADAMTS-13 activity, a longer activated partial thromboplastin time and the need for cardiopulmonary bypass correlated with postoperative bleeding (P < 0.05). Postoperatively, ADAMTS-13 activity increased but less extensively than VWF:Ag (respectively, 2.23 and 2.83 times baseline, P < 0.0001), resulting in an increased VWF:Ag/ADAMTS-13 activity ratio (1.20 to 1.54, respectively, pre- and postoperative median values, P = 0.0029). ADAMTS-13 consumption was further confirmed by decreased ADAMTS-13 antigenic concentration (0.91 ± 0.30 to 0.70 ± 0.25 µg/mL, P < 0.0001) and persistent proteolysis of VWF multimers. We conclude that, in pediatric CCHD, changes in circulating ADAMTS-13 suggest enzyme consumption, associated with abnormal structure and function of VWF. PMID:23558858

  10. Neprilysins: an evolutionarily conserved family of metalloproteases that play important roles in reproduction in Drosophila.

    PubMed

    Sitnik, Jessica L; Francis, Carmen; Hens, Korneel; Huybrechts, Roger; Wolfner, Mariana F; Callaerts, Patrick

    2014-03-01

    Members of the M13 class of metalloproteases have been implicated in diseases and in reproductive fitness. Nevertheless, their physiological role remains poorly understood. To obtain a tractable model with which to analyze this protein family's function, we characterized the gene family in Drosophila melanogaster and focused on reproductive phenotypes. The D. melanogaster genome contains 24 M13 class protease homologs, some of which are orthologs of human proteases, including neprilysin. Many are expressed in the reproductive tracts of either sex. Using RNAi we individually targeted the five Nep genes most closely related to vertebrate neprilysin, Nep1-5, to investigate their roles in reproduction. A reduction in Nep1, Nep2, or Nep4 expression in females reduced egg laying. Nep1 and Nep2 are required in the CNS and the spermathecae for wild-type fecundity. Females that are null for Nep2 also show defects as hosts of sperm competition as well as an increased rate of depletion for stored sperm. Furthermore, eggs laid by Nep2 mutant females are fertilized normally, but arrest early in embryonic development. In the male, only Nep1 was required to induce normal patterns of female egg laying. Reduction in the expression of Nep2-5 in the male did not cause any dramatic effects on reproductive fitness, which suggests that these genes are either nonessential for male fertility or perform redundant functions. Our results suggest that, consistent with the functions of neprilysins in mammals, these proteins are also required for reproduction in Drosophila, opening up this model system for further functional analysis of this protein class and their substrates.

  11. Role in proinflammatory response of YghJ, a secreted metalloprotease from neonatal septicemic Escherichia coli.

    PubMed

    Tapader, Rima; Bose, Dipro; Basu, Pallabi; Mondal, Moumita; Mondal, Ayan; Chatterjee, Nabendu Sekhar; Dutta, Pujarini; Basu, Sulagna; Bhadra, Rupak K; Pal, Amit

    2016-11-01

    Neonatal sepsis is the invasion of microbial pathogens into blood stream and is associated with a systemic inflammatory response with production and release of a wide range of inflammatory mediators. The increased serum levels of cytokines were found to correlate with the severity and mortality in course of sepsis. There have been no reports on the role of microbial proteases in stimulation of proinflammatory response in neonatal sepsis. We have identified YghJ, a secreted metalloprotease from a neonatal septicemic Escherichia coli (NSEC) isolate. The protease was partially purified from culture supernatant by successive anion and gel filtration chromatography. MS/MS peptide sequencing of the protease showed homology with YghJ. YghJ was cloned, expressed and purified in pBAD TOPO expression vector. YghJ was found to be proteolytically active against Methoxysuccinyl Ala-Ala-Pro-Met-p-nitroanilide oligopeptide substrate, but not against casein and gelatin. YghJ showed optimal activity at pH 7-8 and at temperatures 37-40°C. YghJ showed clear changes in cellular morphologies of Int407, HT-29 and HEK293 cells. YghJ stimulated the secretion of cytokines IL-1α, IL-1β and TNF-α in murine macrophages (RAW 264.7) and IL-8 from human intestinal epithelial cells (HT-29). YghJ also down-regulated the production of anti-inflammatory cytokines such as IL-10. YghJ is present in both septicemic (78%) and fecal E. coli isolates (54%). However, expression and secretion of YghJ is significantly higher among the septicemic (89%) than the fecal isolates (33%). This is the first study to show the role of a microbial protease, YghJ in triggering proinflammatory response in NSEC.

  12. Identification, cloning, expression and functional characterization of an astacin-like metalloprotease toxin from Loxosceles intermedia (brown spider) venom

    PubMed Central

    da Silveira, Rafael B.; Wille, Ana C. M.; Chaim, Olga M.; Appel, Marcia H.; Silva, Dilza T.; Franco, Célia R. C.; Toma, Leny; Mangili, Oldemir C.; Gremski, Waldemiro; Dietrich, Carl P.; Nader, Helena B.; Veiga, Silvio S.

    2007-01-01

    Injuries caused by brown spiders (Loxosceles genus) are associated with dermonecrotic lesions with gravitational spreading and systemic manifestations. The venom has a complex composition containing many different toxins, of which metalloproteases have been described in many different species of this genus. These toxins may degrade extracellular matrix constituents acting as a spreading factor. By using a cDNA library from an Loxosceles intermedia venom gland, we cloned and expressed a 900 bp cDNA, which encoded a signal peptide and a propeptide, which corresponded to a 30 kDa metalloprotease, now named LALP (Loxosceles astacin-like protease). Recombinant LALP was refolded and used to produce a polyclonal antiserum, which showed cross-reactivity with a 29 kDa native venom protein. CD analysis provided evidence that the recombinant LALP toxin was folded correctly, was still in a native conformation and had not aggregated. LALP addition to endothelial cell cultures resulted in de-adhesion of the cells, and also in the degradation of fibronectin and fibrinogen (this could be inhibited by the presence of the bivalent chelator 1,10-phenanthroline) and of gelatin in vitro. Sequence comparison (nucleotide and deduced amino acid), phylogenetic analysis and analysis of the functional recombinant toxin revealed that LALP is related in both structure and function to the astacin family of metalloproteases. This suggests that an astacin-like toxin is present in a animal venom secretion and indicates that recombinant LALP will be a useful tool for future structural and functional studies on venom and the astacin family. PMID:17535156

  13. Identification, cloning, expression and functional characterization of an astacin-like metalloprotease toxin from Loxosceles intermedia (brown spider) venom.

    PubMed

    da Silveira, Rafael B; Wille, Ana C M; Chaim, Olga M; Appel, Marcia H; Silva, Dilza T; Franco, Célia R C; Toma, Leny; Mangili, Oldemir C; Gremski, Waldemiro; Dietrich, Carl P; Nader, Helena B; Veiga, Silvio S

    2007-09-01

    Injuries caused by brown spiders (Loxosceles genus) are associated with dermonecrotic lesions with gravitational spreading and systemic manifestations. The venom has a complex composition containing many different toxins, of which metalloproteases have been described in many different species of this genus. These toxins may degrade extracellular matrix constituents acting as a spreading factor. By using a cDNA library from an Loxosceles intermedia venom gland, we cloned and expressed a 900 bp cDNA, which encoded a signal peptide and a propeptide, which corresponded to a 30 kDa metalloprotease, now named LALP (Loxosceles astacin-like protease). Recombinant LALP was refolded and used to produce a polyclonal antiserum, which showed cross-reactivity with a 29 kDa native venom protein. CD analysis provided evidence that the recombinant LALP toxin was folded correctly, was still in a native conformation and had not aggregated. LALP addition to endothelial cell cultures resulted in de-adhesion of the cells, and also in the degradation of fibronectin and fibrinogen (this could be inhibited by the presence of the bivalent chelator 1,10-phenanthroline) and of gelatin in vitro. Sequence comparison (nucleotide and deduced amino acid), phylogenetic analysis and analysis of the functional recombinant toxin revealed that LALP is related in both structure and function to the astacin family of metalloproteases. This suggests that an astacin-like toxin is present in a animal venom secretion and indicates that recombinant LALP will be a useful tool for future structural and functional studies on venom and the astacin family.

  14. Solution structure and dynamics of C-terminal regulatory domain of Vibrio vulnificus extracellular metalloprotease

    SciTech Connect

    Yun, Ji-Hye; Kim, Heeyoun; Park, Jung Eun; Lee, Jung Sup; Lee, Weontae

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer We have determined solution structures of vEP C-terminal regulatory domain. Black-Right-Pointing-Pointer vEP C-ter100 has a compact {beta}-barrel structure with eight anti-parallel {beta}-strands. Black-Right-Pointing-Pointer Solution structure of vEP C-ter100 shares its molecular topology with that of the collagen-binding domain of collagenase. Black-Right-Pointing-Pointer Residues in the {beta}3 region of vEP C-ter100 might be important in putative ligand/receptor binding. Black-Right-Pointing-Pointer vEP C-ter100 interacts strongly with iron ion. -- Abstract: An extracellular metalloprotease (vEP) secreted by Vibrio vulnificus ATCC29307 is a 45-kDa proteolytic enzyme that has prothrombin activation and fibrinolytic activities during bacterial infection. The action of vEP could result in clotting that could serve to protect the bacteria from the host defense machinery. Very recently, we showed that the C-terminal propeptide (C-ter100), which is unique to vEP, is involved in regulation of vEP activity. To understand the structural basis of this function of vEP C-ter100, we have determined the solution structure and backbone dynamics using multidimensional nuclear magnetic resonance spectroscopy. The solution structure shows that vEP C-ter100 is composed of eight anti-parallel {beta}-strands with a unique fold that has a compact {beta}-barrel formation which stabilized by hydrophobic and hydrogen bonding networks. Protein dynamics shows that the overall structure, including loops, is very rigid and stabilized. By structural database analysis, we found that vEP C-ter100 shares its topology with that of the collagen-binding domain of collagenase, despite low sequence homology between the two domains. Fluorescence assay reveals that vEP C-ter100 interacts strongly with iron (Fe{sup 3+}). These findings suggest that vEP protease might recruit substrate molecules, such as collagen, by binding at C-ter100 and that vEP participates

  15. Unusual life cycle and impact on microfibril assembly of ADAMTS17, a secreted metalloprotease mutated in genetic eye disease

    PubMed Central

    Hubmacher, Dirk; Schneider, Michael; Berardinelli, Steven J.; Takeuchi, Hideyuki; Willard, Belinda; Reinhardt, Dieter P.; Haltiwanger, Robert S.; Apte, Suneel S.

    2017-01-01

    Secreted metalloproteases have diverse roles in the formation, remodeling, and the destruction of extracellular matrix. Recessive mutations in the secreted metalloprotease ADAMTS17 cause ectopia lentis and short stature in humans with Weill-Marchesani-like syndrome and primary open angle glaucoma and ectopia lentis in dogs. Little is known about this protease or its connection to fibrillin microfibrils, whose major component, fibrillin-1, is genetically associated with ectopia lentis and alterations in height. Fibrillin microfibrils form the ocular zonule and are present in the drainage apparatus of the eye. We show that recombinant ADAMTS17 has unique characteristics and an unusual life cycle. It undergoes rapid autocatalytic processing in trans after its secretion from cells. Secretion of ADAMTS17 requires O-fucosylation and its autocatalytic activity does not depend on propeptide processing by furin. ADAMTS17 binds recombinant fibrillin-2 but not fibrillin-1 and does not cleave either. It colocalizes to fibrillin-1 containing microfibrils in cultured fibroblasts and suppresses fibrillin-2 (FBN2) incorporation in microfibrils, in part by transcriptional downregulation of Fbn2 mRNA expression. RNA in situ hybridization detected Adamts17 expression in specific structures in the eye, skeleton and other organs, where it may regulate the fibrillin isoform composition of microfibrils. PMID:28176809

  16. Systematic substrate identification indicates a central role for the metalloprotease ADAM10 in axon targeting and synapse function

    PubMed Central

    Kuhn, Peer-Hendrik; Colombo, Alessio Vittorio; Schusser, Benjamin; Dreymueller, Daniela; Wetzel, Sebastian; Schepers, Ute; Herber, Julia; Ludwig, Andreas; Kremmer, Elisabeth; Montag, Dirk; Müller, Ulrike; Schweizer, Michaela; Saftig, Paul; Bräse, Stefan; Lichtenthaler, Stefan F

    2016-01-01

    Metzincin metalloproteases have major roles in intercellular communication by modulating the function of membrane proteins. One of the proteases is the a-disintegrin-and-metalloprotease 10 (ADAM10) which acts as alpha-secretase of the Alzheimer's disease amyloid precursor protein. ADAM10 is also required for neuronal network functions in murine brain, but neuronal ADAM10 substrates are only partly known. With a proteomic analysis of Adam10-deficient neurons we identified 91, mostly novel ADAM10 substrate candidates, making ADAM10 a major protease for membrane proteins in the nervous system. Several novel substrates, including the neuronal cell adhesion protein NrCAM, are involved in brain development. Indeed, we detected mistargeted axons in the olfactory bulb of conditional ADAM10-/- mice, which correlate with reduced cleavage of NrCAM, NCAM and other ADAM10 substrates. In summary, the novel ADAM10 substrates provide a molecular basis for neuronal network dysfunctions in conditional ADAM10-/- mice and demonstrate a fundamental function of ADAM10 in the brain. DOI: http://dx.doi.org/10.7554/eLife.12748.001 PMID:26802628

  17. Ancylostoma caninum MTP-1, an astacin-like metalloprotease secreted by infective hookworm larvae, is involved in tissue migration.

    PubMed

    Williamson, Angela L; Lustigman, Sara; Oksov, Yelena; Deumic, Vehid; Plieskatt, Jordan; Mendez, Susana; Zhan, Bin; Bottazzi, Maria Elena; Hotez, Peter J; Loukas, Alex

    2006-02-01

    Infective larvae (L3) of nematodes secrete macromolecules that are critical to infection and establishment of the parasite in the host. The dog hookworm Ancylostoma caninum secretes an astacin-like metalloprotease, Ac-MTP-1, upon activation in vitro with host serum. Recombinant Ac-MTP-1 was expressed in the baculovirus/insect cell system as a secreted protein and was purified from culture medium by two separate methods, cation-exchange fast-performance liquid chromatography and gelatin-affinity chromatography. Recombinant MTP-1 was catalytically active and digested a range of native and denatured connective tissue substrates, including gelatin, collagen, laminin, and fibronectin. A dog was immunized with recombinant Ac-MTP-1 formulated with AS03 adjuvant, and the antiserum was used to immunolocalize the anatomic sites of expression within A. caninum L3 to secretory granules in the glandular esophagus and the channels that connect the esophagus to the L3 surface and to the cuticle. Antiserum inhibited the ability of recombinant MTP-1 to digest collagen by 85% and inhibited larval migration through tissue in vitro by 70 to 75%, in contrast to just 5 to 10% inhibition obtained with preimmunization serum. The metalloprotease inhibitors EDTA and 1,10-phenanthroline also reduced the penetration of L3 through skin in vitro by 43 to 61%. The data strongly suggest that Ac-MTP-1 is critical for the invasion process of hookworm larvae, and moreover, that antibodies against the enzyme can neutralize its function and inhibit migration.

  18. Purification and characterisation of κ-casein specific milk-clotting metalloprotease from Termitomyces clypeatus MTCC 5091.

    PubMed

    Majumder, Rajib; Banik, Samudra Prosad; Khowala, Suman

    2015-04-15

    Milk-clotting enzymes are valued as chymosin-like protease substitutes for cheese making industries. An extracellular metalloprotease (AcPs) with high milk-clotting activity was purified from edible mushroom Termitomyces clypeatus and characterised. AcPs was preferentially active towards κ-casein, analysed by Urea-PAGE and LC-ESI-MS, whereas the degradation of α and β-casein components by AcPs proceeded slowly justifying its suitability for cheese making. RP-HPLC peptide profiling revealed that the AcPs activity on milk casein was similar to that of a commercial milk coagulant. The enzyme exhibited pH and temperature optima at 5.0 and 45 °C, respectively and showed a pI value of 4.6. One- and two dimensional zymographies revealed a single polypeptide band with proteolytic signal. The MALDI-TOF/MS followed by peptide mass fingerprinting revealed homology with a predicted protein of Populus trichocarpa. To our knowledge, this is the first report on a metalloprotease from T. clypeatus, and the results indicate that this enzyme can be considered as a potential substitute for chymosin in cheese manufacturing.

  19. The astacin metalloprotease moulting enzyme NAS-36 is required for normal cuticle ecdysis in free-living and parasitic nematodes.

    PubMed

    Stepek, Gillian; McCormack, Gillian; Birnie, Andrew J; Page, Antony P

    2011-02-01

    Nematodes represent one of the most abundant and species-rich groups of animals on the planet, with parasitic species causing chronic, debilitating infections in both livestock and humans worldwide. The prevalence and success of the nematodes is a direct consequence of the exceptionally protective properties of their cuticle. The synthesis of this cuticle is a complex multi-step process, which is repeated 4 times from hatchling to adult and has been investigated in detail in the free-living nematode, Caenorhabditis elegans. This process is known as moulting and involves numerous enzymes in the synthesis and degradation of the collagenous matrix. The nas-36 and nas-37 genes in C. elegans encode functionally conserved enzymes of the astacin metalloprotease family which, when mutated, result in a phenotype associated with the late-stage moulting defects, namely the inability to remove the preceding cuticle. Extensive genome searches in the gastrointestinal nematode of sheep, Haemonchus contortus, and in the filarial nematode of humans, Brugia malayi, identified NAS-36 but not NAS-37 homologues. Significantly, the nas-36 gene from B. malayi could successfully complement the moult defects associated with C. elegans nas-36, nas-37 and nas-36/nas-37 double mutants, suggesting a conserved function for NAS-36 between these diverse nematode species. This conservation between species was further indicated when the recombinant enzymes demonstrated a similar range of inhibitable metalloprotease activities.

  20. Tumor necrosis factor-α alters integrins and metalloprotease ADAM12 levels and signaling in differentiating myoblasts.

    PubMed

    Grzelkowska-Kowalczyk, K; Tokarska, J; Grabiec, K; Gajewska, M; Milewska, M; Błaszczyk, M

    2016-01-01

    The extracellular matrix (ECM) is important in the regulation of myogenesis. We hypothesized that tumor necrosis factor-α (TNF-α) modifies ECM during differentiation of mouse C2C12 myoblasts. Exogenous TNF-α (1 ng/ml) stimulated myoblast fusion on the 3rd day (by 160% vs control) but not on the 5th day of myogenesis. The level of integrin α5 was significantly augmented by TNF-α during 5 day-differentiation; however, integrin β1 was higher than control only on the 3rd day of cytokine treatment. Both the abundance of integrin α5 bound to actin and the level of integrin β1 complexed with integrin α5 increased in the presence of TNF-α, especially on the 3rd day of differentiation. Similarly, the stimulatory effects of TNF-α on integrin α3, metalloprotease ADAM12 and kinases related to integrins, FAK and ILK, were limited to the 3rd day of differentiation. We concluded that TNF-α-induced changes in ECM components in differentiating myogenic cells, i.e. i) increased expression of integrin α5, β1, α3, and metalloprotease ADAM12, ii) enhanced formation of α5β1 integrin receptors and interaction of integrin α5-cytoskeleton, and iii) increased expression of kinases associated with integrin signaling, FAK and ILK, were temporarily associated with the onset of myocyte fusion.

  1. Hampered cumulus expansion of porcine cumulus-oocyte complexes by excessive presence of alpha2 -macroglobulin is likely mediated via inhibition of zinc-dependent metalloproteases.

    PubMed

    Appeltant, Ruth; Beek, Josine; Maes, Dominiek; Bijttebier, Jo; Van Steendam, Katleen; Nauwynck, Hans; Van Soom, Ann

    2017-01-26

    In vitro maturation (IVM) in serum causes hampered expansion of porcine cumulus-oocyte complexes (COCs) due to excessive alpha2 -macroglobulin (A2M). This study investigated two hypotheses that could explain the effect of A2M: (i) binding of epidermal growth factor (EGF) to A2M, followed by its decreased availability; and (ii) inhibition of zinc-dependent metalloproteases. Cumulus expansion was evaluated based on the diameter of the COCs, the proportion of COCs participating in a floating cloud and the proportion of COCs with loss of cumulus cells. The first hypothesis of decreased EGF availability was tested by increasing the EGF concentration (20 and 50 ng/mL vs. 10 ng/mL), but was not confirmed because cumulus expansion did not improve. To verify the second hypothesis of inhibited zinc-dependent metalloproteases, the effect of tissue inhibitor of metalloproteases-3 (TIMP-3) on cumulus expansion during IVM with and without A2M was investigated. To immuno-neutralize A2M, serum was pre-incubated with A2M antibodies. Impaired cumulus expansion because of TIMP-3 could only be observed during IVM in 10% of serum with A2M antibodies. No effect of TIMP-3 was observed in medium without A2M antibodies. These results indicate that A2M and TIMP-3 share a common target, a zinc-dependent metalloprotease. Future research is directed toward the identification of the protease involved.

  2. Activation of kinin B1 receptor increases the release of metalloproteases-2 and -9 from both estrogen-sensitive and -insensitive breast cancer cells.

    PubMed

    Ehrenfeld, Pamela; Conejeros, Ivan; Pavicic, Maria F; Matus, Carola E; Gonzalez, Carlos B; Quest, Andrew F G; Bhoola, Kanti D; Poblete, Maria T; Burgos, Rafael A; Figueroa, Carlos D

    2011-02-01

    The kinin B(1) receptor (B(1)R) agonist Lys-des[Arg(9)]-bradykinin (LDBK) increases proliferation of estrogen-sensitive breast cancer cells by a process involving activation of the epidermal growth factor receptor (EGFR) and downstream signaling via the ERK1/2 mitogen-activated protein kinase pathway. Here, we investigated whether B(1)R stimulation induced release of the extracellular matrix metalloproteases MMP-2 and MMP-9 via ERK-dependent pathway in both estrogen-sensitive MCF-7 and -insensitive MDA-MB-231 breast cancer cells. Cells were stimulated with 1-100nM of the B(1)R agonist for variable time-points. Western blotting and gelatin zymography were used to evaluate the presence of MMP-2 and MMP-9 in the extracellular medium. Stimulation of B(1)R with as little as 1 nM LDBK induced the accumulation of these metalloproteases in the medium within 5-30min of stimulation. In parallel, immunocytochemistry revealed that metalloprotease levels in the breast cancer cells declined after stimulation. This effect was blocked either by pre-treating the cells with a B(1)R antagonist or by transfecting with B(1)R-specific siRNA. Activation of the ERK1/2 pathway and EGFR transactivation was required for release of metalloproteases because both the MEK1 inhibitor, PD98059, and AG1478, an inhibitor of the EGFR-tyrosine kinase activity, blocked this event. The importance of EGFR-dependent signaling was additionally confirmed since transfection of cells with the dominant negative EGFR mutant HERCD533 blocked the release of metalloproteases. Thus, activation of B(1)R is likely to enhance breast cancer cells invasiveness by releasing enzymes that degrade the extracellular matrix and thereby favor metastasis.

  3. A Disintegrin-like and Metalloprotease (Reprolysin-type) with Thrombospondin Type 1 Motif (ADAMTS) Superfamily: Functions and Mechanisms*

    PubMed Central

    Apte, Suneel S.

    2009-01-01

    Together with seven ADAMTS-like proteins, the 19 mammalian ADAMTS proteases constitute a superfamily. ADAMTS proteases are secreted zinc metalloproteases whose hallmark is an ancillary domain containing one or more thrombospondin type 1 repeats. ADAMTS-like proteins resemble ADAMTS ancillary domains and lack proteolytic activity. Vertebrate expansion of the superfamily reflects emergence of new substrates, duplication of proteolytic activities in new contexts, and cooperative functions of the duplicated genes. ADAMTS proteases are involved in maturation of procollagen and von Willebrand factor, as well as in extracellular matrix proteolysis relating to morphogenesis, angiogenesis, ovulation, cancer, and arthritis. New insights into ADAMTS mechanisms indicate significant regulatory roles for ADAMTS ancillary domains, propeptide processing, and glycosylation. ADAMTS-like proteins appear to have regulatory roles in the extracellular matrix. PMID:19734141

  4. The Adam family metalloprotease Kuzbanian regulates the cleavage of the roundabout receptor to control axon repulsion at the midline.

    PubMed

    Coleman, Hope A; Labrador, Juan-Pablo; Chance, Rebecca K; Bashaw, Greg J

    2010-07-01

    Slits and their Roundabout (Robo) receptors mediate repulsive axon guidance at the Drosophila ventral midline and in the vertebrate spinal cord. Slit is cleaved to produce fragments with distinct signaling properties. In a screen for genes involved in Slit-Robo repulsion, we have identified the Adam family metalloprotease Kuzbanian (Kuz). Kuz does not regulate midline repulsion through cleavage of Slit, nor is Slit cleavage essential for repulsion. Instead, Kuz acts in neurons to regulate repulsion and Kuz can cleave the Robo extracellular domain in Drosophila cells. Genetic rescue experiments using an uncleavable form of Robo show that this receptor does not maintain normal repellent activity. Finally, Kuz activity is required for Robo to recruit its downstream signaling partner, Son of sevenless (Sos). These observations support the model that Kuz-directed cleavage is important for Robo receptor activation.

  5. Induction of matrix metalloprotease-1 gene expression by retinoic acid in the human pancreatic tumour cell line Dan-G

    PubMed Central

    Marschall, Z von; Riecken, E-O; Rosewicz, S

    1999-01-01

    We have investigated the effects of retinoic acid (RA) on matrix metalloprotease-1 (MMP-1) gene expression in the human pancreatic tumour cell line Dan-G. 13-cis RA results in a time- and dose-dependent increase of MMP-1 protein concentration. These stimulatory effects were paralleled by a time- and dose-dependent increase of MMP-1 mRNA steady-state concentrations. Nuclear run-on analysis revealed that the increase of MMP-1 mRNA was partially due to an increase of MMP-1 gene transcription. In addition, 13-cis RA treatment results in an increase of MMP-1 mRNA stability. These data demonstrate that RA stimulates MMP-1 gene expression in human pancreatic carcinoma cells by transcriptional and post-transcriptional mechanisms. © 1999 Cancer Research Campaign PMID:10362099

  6. PKC/ROS-Mediated NLRP3 Inflammasome Activation Is Attenuated by Leishmania Zinc-Metalloprotease during Infection

    PubMed Central

    Jung, Jee Yong; Chang, Kwang-Poo; Olivier, Martin

    2015-01-01

    Parasites of the Leishmania genus infect and survive within macrophages by inhibiting several microbicidal molecules, such as nitric oxide and pro-inflammatory cytokines. In this context, various species of Leishmania have been reported to inhibit or reduce the production of IL-1β both in vitro and in vivo. However, the mechanism whereby Leishmania parasites are able to affect IL-1β production and secretion by macrophages is still not fully understood. Dependent on the stimulus at hand, the maturation of IL-1β is facilitated by different inflammasome complexes. The NLRP3 inflammasome has been shown to be of pivotal importance in the detection of danger molecules such as inorganic crystals like asbestos, silica and malarial hemozoin, (HZ) as well as infectious agents. In the present work, we investigated whether Leishmania parasites modulate NLRP3 inflammasome activation. Using PMA-differentiated THP-1 cells, we demonstrate that Leishmania infection effectively inhibits macrophage IL-1β production upon stimulation. In this context, the expression and activity of the metalloprotease GP63 - a critical virulence factor expressed by all infectious Leishmania species - is a prerequisite for a Leishmania-mediated reduction of IL-1β secretion. Accordingly, L. mexicana, purified GP63 and GP63-containing exosomes, caused the inhibition of macrophage IL-1β production. Leishmania-dependent suppression of IL-1β secretion is accompanied by an inhibition of reactive oxygen species (ROS) production that has previously been shown to be associated with NLRP3 inflammasome activation. The observed loss of ROS production was due to an impaired PKC-mediated protein phosphorylation. Furthermore, ROS-independent inflammasome activation was inhibited, possibly due to an observed GP63-dependent cleavage of inflammasome and inflammasome-related proteins. Collectively for the first time, we herein provide evidence that the protozoan parasite Leishmania, through its surface

  7. Disintegrin Metalloprotease (ADAM) 10 Regulates Endothelial Permeability and T Cell Transmigration by Proteolysis of Vascular Endothelial Cadherin

    PubMed Central

    Schulz, Beate; Pruessmeyer, Jessica; Maretzky, Thorsten; Ludwig, Andreas; Blobel, Carl P.; Saftig, Paul; Reiss, Karina

    2009-01-01

    Vascular endothelial (VE)-cadherin is the major adhesion molecule of endothelial adherens junctions. It plays an essential role in controlling endothelial permeability, vascular integrity, leukocyte transmigration, and angiogenesis. Elevated levels of soluble VE-cadherin are associated with diseases like coronary atherosclerosis. Previous data showed that the extracellular domain of VE-cadherin is released by an unknown metalloprotease activity during apoptosis. In this study, we used gain of function analyses, inhibitor studies and RNA interference experiments to analyze the proteolytic release of VE-cadherin in human umbilical vein endothelial cells (HUVECs). We found that VE-cadherin is specifically cleaved by the disintegrin and metalloprotease ADAM10 in its ectodomain releasing a soluble fragment and generating a carboxyterminal membrane bound stub, which is a substrate for a subsequent γ-secretase cleavage. This ADAM10-mediated proteolysis could be induced by Ca2+-influx and staurosporine treatment, indicating that ADAM10-mediated VE-cadherin cleavage contributes to the dissolution of adherens junctions during endothelial cell activation and apoptosis, respectively. In contrast, protein kinase C activation or inhibition did not modulate VE-cadherin processing. Increased ADAM10 expression was functionally associated with an increase in endothelial permeability. Remarkably, our data indicate that ADAM10 activity also contributes to the thrombin-induced decrease of endothelial cell-cell adhesion. Moreover, knockdown of ADAM10 in HUVECs as well as in T cells by small interfering RNA impaired T cell transmigration. Taken together our data identify ADAM10 as a novel regulator of vascular permeability and demonstrate a hitherto unknown function of ADAM10 in the regulation of VE-cadherin-dependent endothelial cell functions and leukocyte transendothelial migration. PMID:18420943

  8. TspanC8 Tetraspanins and A Disintegrin and Metalloprotease 10 (ADAM10) Interact via Their Extracellular Regions

    PubMed Central

    Noy, Peter J.; Yang, Jing; Reyat, Jasmeet S.; Matthews, Alexandra L.; Charlton, Alice E.; Furmston, Joanna; Rogers, David A.; Rainger, G. Ed; Tomlinson, Michael G.

    2016-01-01

    A disintegrin and metalloprotease 10 (ADAM10) is a ubiquitously expressed transmembrane metalloprotease that cleaves the extracellular regions from its transmembrane substrates. ADAM10 is essential for embryonic development and is implicated in cancer, Alzheimer, and inflammatory diseases. The tetraspanins are a superfamily of 33 four-transmembrane proteins in mammals, of which the TspanC8 subgroup (Tspan5, 10, 14, 15, 17, and 33) promote ADAM10 intracellular trafficking and enzymatic maturation. However, the interaction between TspanC8s and ADAM10 has only been demonstrated in overexpression systems and the interaction mechanism remains undefined. To address these issues, an antibody was developed to Tspan14, which was used to show co-immunoprecipitation of Tspan14 with ADAM10 in primary human cells. Chimeric Tspan14 constructs demonstrated that the large extracellular loop of Tspan14 mediated its co-immunoprecipitation with ADAM10, and promoted ADAM10 maturation and trafficking to the cell surface. Chimeric ADAM10 constructs showed that membrane-proximal stalk, cysteine-rich, and disintegrin domains of ADAM10 mediated its co-immunoprecipitation with Tspan14 and other TspanC8s. This TspanC8-interacting region was required for ADAM10 exit from the endoplasmic reticulum. Truncated ADAM10 constructs revealed differential TspanC8 binding requirements for the stalk, cysteine-rich, and disintegrin domains. Moreover, Tspan15was the only TspanC8 to promote cleavage of the ADAM10 substrate N-cadherin, whereas Tspan14 was unique in reducing cleavage of the platelet collagen receptor GPVI. These findings suggest that ADAM10 may adopt distinct conformations in complex with different TspanC8s, which could impact on substrate selectivity. Furthermore, this study identifies regions of TspanC8s and ADAM10 for potential interaction-disrupting therapeutic targeting. PMID:26668317

  9. Heightened cleavage of Axl receptor tyrosine kinase by ADAM metalloproteases may contribute to disease pathogenesis in SLE

    PubMed Central

    Orme, Jacob J.; Du, Yong; Vanarsa, Kamala; Mayeux, Jessica; Li, Li; Mutwally, Azza; Arriens, Cristina; Min, Soyoun; Hutcheson, Jack; Davis, Laurie S.; Chong, Benjamin F.; Satterthwaite, Anne B.; Wu, Tianfu; Mohan, Chandra

    2016-01-01

    Systemic lupus erythematosus (SLE) is characterized by antibody-mediated chronic inflammation in the kidney, lung, skin, and other organs to cause inflammation and damage. Several inflammatory pathways are dysregulated in SLE, and understanding these pathways may improve diagnosis and treatment. In one such pathway, Axl tyrosine kinase receptor responds to Gas6 ligand to block inflammation in leukocytes. A soluble form of the Axl receptor ectodomain (sAxl) is elevated in serum from patients with SLE and lupus-prone mice. We hypothesized that sAxl in SLE serum originates from the surface of leukocytes and that the loss of leukocyte Axl contributes to the disease. We determined that macrophages and B cells are a source of sAxl in SLE and in lupus-prone mice. Shedding of the Axl ectodomain from the leukocytes of lupus-prone mice is mediated by the matrix metalloproteases ADAM10 and TACE (ADAM17). Loss of Axl from lupus-prone macrophages renders them unresponsive to Gas6-induced anti-inflammatory signaling in vitro. This phenotype is rescued by combined ADAM10/TACE inhibition. Mice with Axl-deficient macrophages develop worse disease than controls when challenged with anti-glomerular basement membrane (anti-GBM) sera in an induced model of nephritis. ADAM10 and TACE also mediate human SLE PBMC Axl cleavage. Collectively, these studies indicate that increased metalloprotease-mediated cleavage of leukocyte Axl may contribute to end organ disease in lupus. They further suggest dual ADAM10/TACE inhibition as a potential therapeutic modality in SLE. PMID:27237127

  10. Differential metalloprotease content and activity of three Loxosceles spider venoms revealed using two-dimensional electrophoresis approaches.

    PubMed

    Trevisan-Silva, Dilza; Bednaski, Aline Viana; Gremski, Luiza Helena; Chaim, Olga Meiri; Veiga, Silvio Sanches; Senff-Ribeiro, Andrea

    2013-12-15

    Loxosceles bites have been associated with characteristic dermonecrotic lesions with gravitational spreading and systemic manifestations. Venom primarily comprises peptides and protein molecules (5-40 kDa) with multiple biological activities. Although poorly studied, metalloproteases have been identified in venoms of several Loxosceles species, presenting proteolytic effects on extracellular matrix components. The characterization of an Astacin-like protease (LALP) in Loxosceles intermedia venom was the first report of an Astacin family member as a component of animal venom. Recently, these proteases were described as a gene family in L. intermedia, Loxosceles laeta and Loxosceles gaucho. Herein, the whole venom complexity of these three Loxosceles species was analyzed using two-dimensional electrophoresis (2DE). Subproteomes of LALPs were explored through 2DE immunostaining using anti-LALP1 antibodies and 2DE gelatin zymogram. Proteins presented molecular masses ranging from 24 to 29 kDa and the majority of these molecules had basic or neutral isoelectric points (6.89-9.93). Likewise, the measurement of gelatinolytic effects of Loxosceles venom using fluorescein-gelatin showed that the three venoms have distinct proteolytic activities. The metalloprotease fibrinogenolytic activities were also evaluated. All venoms showed fibrinogenolytic activity with different proteolytic effects on Aα and Bβ chains of fibrinogen. The results reported herein suggest that the LALP family is larger than indicated in previously published data and that the complex profile of the gelatinolytic activity reflects their relevance in loxoscelism. Furthermore, our investigation implicates the brown spider venom as a source of Astacin-like proteases for use in loxoscelism studies, cell biology research and biotechnological applications.

  11. Characterization of the Zinc-Containing Metalloprotease Encoded by zpx and Development of a Species-Specific Detection Method for Enterobacter sakazakii▿

    PubMed Central

    Kothary, M. H.; McCardell, B. A.; Frazar, C. D.; Deer, D.; Tall, B. D.

    2007-01-01

    Enterobacter sakazakii causes a severe form of neonatal meningitis that occurs as sporadic cases as well as outbreaks. The disease has been epidemiologically associated with consumption of reconstituted, dried infant formulas. Very little information is available regarding pathogenicity of the organism and production of virulence factors. Clinical and environmental strains were screened for production of factors which have activity against Chinese hamster ovary (CHO) cells in tissue culture. Polymyxin B lysate and sonicate preparations but not culture supernatants from the strains caused “rounding” of CHO cells. Subsequent studies showed that the CHO cell-rounding factor is a proteolytic enzyme that has activity against azocasein. The cell-bound protease was isolated by using a combination of polymyxin B lysis, followed by sonication of cells harvested from tryptone broth. The protease was purified to homogeneity by sequential ammonium sulfate precipitation, gel filtration chromatography with Sephadex G-100, hydrophobic interaction chromatography with phenyl-Sepharose CL-4B, and a second gel filtration with Sephadex G-100. In addition to activity against azocasein, the purified protease also exhibits activity against azocoll and insoluble casein but not elastin. The protease has a molecular weight of 38,000 and an isoelectric point of 4.4. It is heat labile and for maximal activity against azocasein has an optimum temperature of 37°C and a pH range of 5 to 7. Proteolytic activity is inhibited by ortho-phenanthroline and Zincov but is not affected by phenylmethylsulfonyl fluoride, N-ethylmaleimide, and trypsin inhibitors, which demonstrates that the protease is a zinc-containing metalloprotease. The metalloprotease does not hemagglutinate chicken or sheep erythrocytes. Twenty-three to 27 of the first 42 N-terminal amino acid residues of the metalloprotease are identical to proteases produced by Serratia proteamaculans, Pectobacterium carotovorum, and Anabaena

  12. IL-1β-induced matrix metalloproteinase-13 is activated by a disintegrin and metalloprotease-28-regulated proliferation of human osteoblast-like cells

    SciTech Connect

    Ozeki, Nobuaki; Kawai, Rie; Yamaguchi, Hideyuki; Hiyama, Taiki; Kinoshita, Katsue; Hase, Naoko; Nakata, Kazuhiko; Kondo, Ayami; Mogi, Makio; Nakamura, Hiroshi

    2014-04-15

    We reported previously that matrix metalloproteinase (MMP)-13 accelerates bone remodeling in oral periradicular lesions, and indicated a potentially unique role for MMP-13 in wound healing and regeneration of alveolar bone. The ADAM (a disintegrin and metalloprotease) family is a set of multifunctional cell surface and secreted glycoproteins, of which ADAM-28 has been localized in bone and bone-like tissues. In this study, we show that interleukin (IL)-1β induces the expression of MMP-13 and ADAM-28 in homogeneous α7 integrin-positive human skeletal muscle stem cell (α7{sup +}hSMSC)-derived osteoblast-like (α7{sup +}hSMSC-OB) cells, and promotes proliferation while inhibiting apoptosis in these cells. At higher concentrations, however, IL-1β failed to induce the expression of these genes and caused an increase in apoptosis. We further employed ADAM-28 small interfering RNA (siRNA) to investigate whether IL-1β-induced MMP-13 expression is linked to this IL-1β-mediated changes in cell proliferation and apoptosis. Silencing ADAM-28 expression potently suppressed IL-1β-induced MMP-13 expression and activity, decreased cell proliferation and increased apoptosis in α7{sup +}hSMSC-OB cells. In contrast, MMP-13 siRNA had no effect on ADAM-28 expression, suggesting ADAM-28 regulates MMP-13. Exogenous MMP-13 induced α7{sup +}hSMSC-OB cell proliferation and could rescue ADAM-28 siRNA-induced apoptosis, and we found that proMMP-13 is partially cleaved into its active form by ADAM-28 in vitro. Overall, our results suggest that IL-1β-induced MMP-13 expression and changes in cell proliferation and apoptosis in α7{sup +}hSMSC-OB cells are regulated by ADAM-28. - Highlights: • IL-1β induces the MMP-13 and ADAM-28 expression in human osteoblast-like cells. • IL-1β-induced MMP-13 expression increases proliferation and decreased apoptosis. • MMP-13 expression induced by IL-1β is regulated by ADAM-28. • proMMP-13 appears to be cleaved into its active form via

  13. Expression and immunological cross-reactivity of LALP3, a novel astacin-like metalloprotease from brown spider (Loxosceles intermedia) venom.

    PubMed

    Morgon, Adriano M; Belisario-Ferrari, Matheus R; Trevisan-Silva, Dilza; Meissner, Gabriel O; Vuitika, Larissa; Marin, Brenda; Tashima, Alexandre K; Gremski, Luiza H; Gremski, Waldemiro; Senff-Ribeiro, Andrea; Veiga, Silvio S; Chaim, Olga M

    2016-01-01

    Loxosceles spiders' venom comprises a complex mixture of biologically active toxins, mostly consisting of low molecular mass components (2-40 kDa). Amongst, isoforms of astacin-like metalloproteases were identified through transcriptome and proteome analyses. Only LALP1 (Loxosceles Astacin-Like protease 1) has been characterized. Herein, we characterized LALP3 as a novel recombinant astacin-like metalloprotease isoform from Loxosceles intermedia venom. LALP3 cDNA was cloned in pET-SUMO vector, and its soluble heterologous expression was performed using a SUMO tag added to LALP3 to achieve solubility in Escherichia coli SHuffle T7 Express LysY cells, which express the disulfide bond isomerase DsbC. Protein purification was conducted by Ni-NTA Agarose resin and assayed for purity by SDS-PAGE under reducing conditions. Immunoblotting analyses were performed with specific antibodies recognizing LALP1 and whole venom. Western blotting showed linear epitopes from recombinant LALP3 that cross-reacted with LALP1, and dot blotting revealed conformational epitopes with native venom astacins. Mass spectrometry analysis revealed that the recombinant expressed protein is an astacin-like metalloprotease from L. intermedia venom. Furthermore, molecular modeling of LALP3 revealed that this isoform contains the zinc binding and Met-turn motifs, forming the active site, as has been observed in astacins. These data confirmed that LALP3, which was successfully obtained by heterologous expression using a prokaryote system, is a new astacin-like metalloprotease isoform present in L. intermedia venom.

  14. The InhA2 Metalloprotease of Bacillus thuringiensis Strain 407 Is Required for Pathogenicity in Insects Infected via the Oral Route

    PubMed Central

    Fedhila, Sinda; Nel, Patricia; Lereclus, Didier

    2002-01-01

    The entomopathogenic bacterium Bacillus thuringiensis is known to secrete a zinc metalloprotease (InhA) that specifically cleaves antibacterial peptides produced by insect hosts. We identified a second copy of the inhA gene, named inhA2, in B. thuringiensis strain 407 Cry−. The inhA2 gene encodes a putative polypeptide showing 66.2% overall identity with the InhA protein and harboring the zinc-binding domain (HEXXH), which is characteristic of the zinc-requiring metalloproteases. We used a transcriptional inhA2′-lacZ fusion to show that inhA2 expression is induced at the onset of the stationary phase and is overexpressed in a Spo0A minus background. The presence of a reverse Spo0A box in the promoter region of inhA2 suggests that Spo0A directly regulates the transcription of inhA2. To determine the role of the InhA and InhA2 metalloproteases in pathogenesis, we used allelic exchange to isolate single and double mutant strains for the two genes. Spores and vegetative cells of the mutant strains were as virulent as those of the parental strain in immunized Bombyx mori larvae infected by the intrahemocoelic route. Exponential phase cells of all the strains displayed the same in vitro potential for colonizing the vaccinated hemocoel. We investigated the synergistic effect of the mutant strain spores on the toxicity of Cry1C proteins against Galleria mellonella larvae infected via the oral pathway. The spores of ΔinhA2 mutant strain were ineffective in providing synergism whereas those of the ΔinhA mutant strain were not. These results indicate that the B. thuringiensis InhA2 zinc metalloprotease has a vital role in virulence when the host is infected via the oral route. PMID:12029046

  15. Epidermal growth factor (EGF) ligand release by substrate-specific a disintegrin and metalloproteases (ADAMs) involves different protein kinase C (PKC) isoenzymes depending on the stimulus.

    PubMed

    Dang, Michelle; Dubbin, Karen; D'Aiello, Antonio; Hartmann, Monika; Lodish, Harvey; Herrlich, Andreas

    2011-05-20

    The dysregulation of EGF family ligand cleavage has severe consequences for the developing as well as the adult organism. Therefore, their production is highly regulated. The limiting step is the ectodomain cleavage of membrane-bound precursors by one of several a disintegrin and metalloprotease (ADAM) metalloproteases, and understanding the regulation of cleavage is an important goal of current research. We have previously reported that in mouse lung epithelial cells, the pro-EGF ligands TGFα, neuregulin 1β (NRG), and heparin-binding EGF are differentially cleaved depending on the cleavage stimulus (Herrlich, A., Klinman, E., Fu, J., Sadegh, C., and Lodish, H. (2008) FASEB J.). In this study in mouse embryonic fibroblasts that lack different ADAMs, we show that induced cleavage of EGF ligands can involve the same substrate-specific metalloprotease but does require different stimulus-dependent signaling pathways. Cleavage was stimulated by phorbol ester (12-O-tetradecanoylphorbol-13-acetate (TPA), a mimic of diacylglycerol and PKC activator), hypertonic stress, lysophosphatidic acid (LPA)-induced G protein-coupled receptor activation, or by ionomycin-induced intracellular calcium release. Although ADAMs showed substrate preference (ADAM17, TGFα and heparin-binding EGF; and ADAM9, NRG), substrate cleavage differed substantially with the stimulus, and cleavage of the same substrate depended on the presence of different, sometimes multiple, PKC isoforms. For instance, classical PKC was required for TPA-induced but not hypertonic stress-induced cleavage of all EGF family ligands. Inhibition of PKCζ enhanced NRG release upon TPA stimulation, but it blocked NRG release in response to hypertonic stress. Our results suggest a model in which substantial regulation of ectodomain cleavage occurs not only on the metalloprotease level but also on the level of the substrate or of a third protein.

  16. Roles of the signal peptide and mature domains in the secretion and maturation of the neutral metalloprotease from Streptomyces cacaoi.

    PubMed Central

    Chang, S C; Su, M H; Lee, Y H

    1997-01-01

    The neutral metalloprotease (Npr) of Streptomyces cacaoi is synthesized as a prepro-Npr precursor form consisting of a secretory signal peptide, a propeptide and the mature metalloprotease. The maturation of Npr occurs extracellularly via an autoproteolytic processing of the secreted pro-Npr. The integrity of the propeptide is essential for the formation of mature active Npr but not for its secretion [Chang, Chang and Lee (1994) J. Biol. Chem. 269, 3548-3554]. In this study we investigated whether the secretion and maturation of Npr require the integrity of its signal peptide region and mature protease domain. Five signal peptide mutants were generated, including the substitution mutations at the positively charged region (mutant IR6LE), the central hydrophobic region (mutants GI19EL and G19N), the boundary of the hydrophobic core-cleavage region (mutant P30L) and at the residues adjacent to the signal peptidase cleavage site (mutant YA33SM). All these lesions delayed the export of Npr to the growth medium and also resulted in a 2-10-fold decrease in Npr export. The most severe effect was noted in mutants GI19EL and P30L. When these signal peptide mutations were fused separately with the propeptide lacking the Npr mature domain, the secretory defect on the propeptide was also observed, and this impairment was again more severely expressed in mutants GI19EL and P30L. Thus the Npr signal peptide seems to have more constraints on the hydrophobic core region and at the proline residue within the boundary of the hydrophobic core-cleavage site. Deletion mutations within the C-terminal mature protease domain that left its active site intact still blocked the proteolytic processing of mutant precursor forms of pro-Npr, although their secretions were unaffected. These results, together with our previous findings, strongly suggest that the signal peptide of Npr plays a pivotal role in the secretion of both Npr and the propeptide, but not in the maturation of Npr. On the

  17. Increased cerebral matrix metalloprotease-9 activity is associated with compromised recovery in the diabetic db/db mouse following a stroke.

    PubMed

    Kumari, Rashmi; Willing, Lisa B; Patel, Shyama D; Baskerville, Karen A; Simpson, Ian A

    2011-12-01

    Diabetes is a major risk factor of stroke and is associated with increased frequency of stroke and a poorer prognosis for recovery. In earlier studies we have utilized type 2 diabetic mouse models of stroke and demonstrated that diabetic db/db and ob/ob mice experience larger infarct volumes and impaired recovery associated with greater infiltration of macrophage following hypoxic-ischemic (H/I) insult than their heterozygous non-diabetic db/+ and ob/+ littermates. To obtain a better understanding of the pathogenesis of the impaired recovery, we have investigated the role of matrix metalloproteases and their endogenous inhibitors in the breakdown of the blood-brain barrier (BBB) following H/I. Diabetic db/db mice showed a significant and more rapid increase in matrix metalloprotease (MMP)-9 mRNA, protein and gelatinolytic activity compared with db/+, which resulted in an increased degradation of occludin and collagen IV and subsequently, an increased BBB permeability and greater infiltration of neutrophils into the infarct area. The expression of the MMPs, especially in the db/+ mice, is preceded by an elevated expression of their endogenous tissue inhibitors of metalloproteases (TIMPs) 1, 2, and 3, whereas in the db/db mice, a lower expression of the TIMPs is associated with greater MMP 3 and 9 expression. These results suggest that an imbalance in the MMPs/TIMPs cascade in the diabetic mouse, particularly MMP-9, results in a greater neutrophil invasion, a compromised BBB and consequently a greater insult.

  18. Metalloprotease-mediated tumor cell shedding of B7-H6, the ligand of the natural killer cell-activating receptor NKp30.

    PubMed

    Schlecker, Eva; Fiegler, Nathalie; Arnold, Annette; Altevogt, Peter; Rose-John, Stefan; Moldenhauer, Gerhard; Sucker, Antje; Paschen, Annette; von Strandmann, Elke Pogge; Textor, Sonja; Cerwenka, Adelheid

    2014-07-01

    Natural killer (NK) cells are potent immune effector cells capable of mediating antitumor responses. Thus, during immunoediting, tumor cell populations evolve strategies to escape NK-cell-mediated recognition. In this study, we report a novel mechanism of immune escape involving tumor cell shedding of B7-H6, a ligand for the activating receptor NKp30 that mediates NK-cell binding and NK-cell-mediated killing. Tumor cells from different cancer entities released B7-H6 by ectodomain shedding mediated by the cell surface proteases "a disintegrin and metalloproteases" (ADAM)-10 and ADAM-17, as demonstrated through the use of pharmacologic inhibitors or siRNA-mediated gene attenuation. Inhibiting this proteolytic shedding process increased the levels of B7-H6 expressed on the surface of tumor cells, enhancing NKp30-mediated activation of NK cells. Notably, we documented elevated levels of soluble B7-H6 levels in blood sera obtained from a subset of patients with malignant melanoma, compared with healthy control individuals, along with evidence of elevated B7-H6 expression in melanoma specimens in situ. Taken together, our results illustrated a novel mechanism of immune escape in which tumor cells impede NK-mediated recognition by metalloprotease-mediated shedding of B7-H6. One implication of our findings is that therapeutic inhibition of specific metalloproteases may help support NK-cell-based cancer therapy.

  19. The NTR module: domains of netrins, secreted frizzled related proteins, and type I procollagen C-proteinase enhancer protein are homologous with tissue inhibitors of metalloproteases.

    PubMed Central

    Bányai, L.; Patthy, L.

    1999-01-01

    Using homology search, structure prediction, and structural characterization methods we show that the C-terminal domains of (1) netrins, (2) complement proteins C3, C4, C5, (3) secreted frizzled-related proteins, and (4) type I procollagen C-proteinase enhancer proteins (PCOLCEs) are homologous with the N-terminal domains of (5) tissue inhibitors of metalloproteinases (TIMPs). The proteins harboring this netrin module (NTR module) fulfill diverse biological roles ranging from axon guidance, regulation of Wnt signaling, to the control of the activity of metalloproteases. With the exception of TIMPs, it is not known at present what role the NTR modules play in these processes. In view of the fact that the NTR modules of TIMPs are involved in the inhibition of matrixin-type metalloproteases and that the NTR module of PCOLCEs is involved in the control of the activity of the astacin-type metalloprotease BMP1, it seems possible that interaction with metzincins could be a shared property of NTR modules and could be critical for the biological roles of the host proteins. PMID:10452607

  20. Structural Basis of Proline-Proline Peptide Bond Specificity of the Metalloprotease Zmp1 Implicated in Motility of Clostridium difficile.

    PubMed

    Schacherl, Magdalena; Pichlo, Christian; Neundorf, Ines; Baumann, Ulrich

    2015-09-01

    Clostridium difficile is a pathogenic bacterium causing gastrointestinal diseases from mild diarrhea to toxic megacolon. In common with other pathogenic bacteria, C. difficile secretes proteins involved in adhesion, colonization, and dissemination. The recently identified Zmp1 is an extracellular metalloprotease showing a unique specificity for Pro-Pro peptide bonds. The endogenous substrates of Zmp1 are two surface proteins implicated in adhesion of C. difficile to surface proteins of human cells. Thus, Zmp1 is believed to be involved in the regulation of the adhesion-motility balance of C. difficile. Here, we report crystal structures of Zmp1 from C. difficile in its unbound and peptide-bound forms. The structure analysis revealed a fold similar to Bacillus anthracis lethal factor. Crystal structures in the open and closed conformation of the S-loop shed light on the mode of binding of the substrate, and reveal important residues for substrate recognition and the strict specificity of Zmp1 for Pro-Pro peptide bonds.

  1. (99m)Tc-MAG3-aptamer for imaging human tumors associated with high level of matrix metalloprotease-9.

    PubMed

    Da Rocha Gomes, Sonia; Miguel, Julie; Azéma, Laurent; Eimer, Sandrine; Ries, Colette; Dausse, Eric; Loiseau, Hugues; Allard, Michèle; Toulmé, Jean-Jacques

    2012-11-21

    The human matrix metalloprotease 9 (hMMP-9) is involved in many physiological processes such as tissue remodeling. Its overexpression in tumors promotes the release of cancer cells thus contributing to tumor metastasis. It is a relevant marker of malignant tumors. We selected an RNA aptamer containing 2'-fluoro, pyrimidine ribonucleosides, that exhibits a strong affinity for hMMP-9 (K(d) = 20 nM) and that discriminates other human MMPs: no binding was detected to either hMMP-2 or -7. Investigating the binding properties of different MMP-9 aptamer variants by surface plasmon resonance allowed the determination of recognition elements. As a result, a truncated aptamer, 36 nucleotides long, was made fully resistant to nuclease following the substitution of every purine ribonucleoside residue by 2'-O-methyl analogues and was conjugated to S-acetylmercaptoacetyltriglycine for imaging purposes. The resulting modified aptamer retained the binding properties of the originally selected sequence. Following (99m)Tc labeling, this aptamer was used for ex vivo imaging slices of human brain tumors. We were able to specifically detect the presence of hMMP-9 in such tissues.

  2. The Apoptogenic Toxin AIP56 Is a Metalloprotease A-B Toxin that Cleaves NF-κb P65

    PubMed Central

    Silva, Daniela S.; Pereira, Liliana M. G.; Moreira, Ana R.; Ferreira-da-Silva, Frederico; Brito, Rui M.; Faria, Tiago Q.; Zornetta, Irene; Montecucco, Cesare; Oliveira, Pedro; Azevedo, Jorge E.; Pereira, Pedro J. B.; Macedo-Ribeiro, Sandra; do Vale, Ana; dos Santos, Nuno M. S.

    2013-01-01

    AIP56 (apoptosis-inducing protein of 56 kDa) is a major virulence factor of Photobacterium damselae piscicida (Phdp), a Gram-negative pathogen that causes septicemic infections, which are among the most threatening diseases in mariculture. The toxin triggers apoptosis of host macrophages and neutrophils through a process that, in vivo, culminates with secondary necrosis of the apoptotic cells contributing to the necrotic lesions observed in the diseased animals. Here, we show that AIP56 is a NF-κB p65-cleaving zinc-metalloprotease whose catalytic activity is required for the apoptogenic effect. Most of the bacterial effectors known to target NF-κB are type III secreted effectors. In contrast, we demonstrate that AIP56 is an A-B toxin capable of acting at distance, without requiring contact of the bacteria with the target cell. We also show that the N-terminal domain cleaves NF-κB at the Cys39-Glu40 peptide bond and that the C-terminal domain is involved in binding and internalization into the cytosol. PMID:23468618

  3. The Role of SnoN in Transforming Growth Factor β1-induced Expression of Metalloprotease-Disintegrin ADAM12*

    PubMed Central

    Solomon, Emilia; Li, Hui; Duhachek Muggy, Sara; Syta, Emilia; Zolkiewska, Anna

    2010-01-01

    Increased expression of metalloprotease-disintegrin ADAM12 is a hallmark of several pathological conditions, including cancer, cardiovascular disease, and certain inflammatory diseases of the central nervous system or the muscoskeletal system. We show that transforming growth factor β1 (TGFβ1) is a potent inducer of ADAM12 mRNA and protein in mouse fibroblasts and in mouse and human mammary epithelial cells. Induction of ADAM12 is detected within 2 h of treatment with TGFβ1, is Smad2/Smad3-dependent, and is a result of derepression of the Adam12 gene. SnoN, a negative regulator of the TGFβ signaling pathway, is a master regulator of ADAM12 expression in response to TGFβ1 stimulation. Overexpression of SnoN in NIH3T3 cells reduces the magnitude of ADAM12 induction by TGFβ1 treatment. Down-regulation of SnoN expression by short hairpin RNA enhances TGFβ1-induced expression of ADAM12. In a panel of TGFβ1-responsive cancer cell lines with high expression of SnoN, induction of ADAM12 by TGFβ1 is significantly impaired, suggesting that the endogenous SnoN plays a role in regulating ADAM12 expression in response to TGFβ1. Identification of SnoN as a repressor of the ADAM12 gene should contribute to advances in the studies on the role of ADAM12 in tumor progression and in the development of other pathologies. PMID:20457602

  4. Cyclosporin A Impairs the Secretion and Activity of ADAMTS13 (A Disintegrin and Metalloprotease with Thrombospondin Type 1 Repeat)*

    PubMed Central

    Hershko, Klilah; Simhadri, Vijaya L.; Blaisdell, Adam; Hunt, Ryan C.; Newell, Jordan; Tseng, Sandra C.; Hershko, Alon Y.; Choi, Jae Won; Sauna, Zuben E.; Wu, Andrew; Bram, Richard J.; Komar, Anton A.; Kimchi-Sarfaty, Chava

    2012-01-01

    The protease ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 repeat) cleaves multimers of von Willebrand factor, thus regulating platelet aggregation. ADAMTS13 deficiency leads to the fatal disorder thrombotic thrombocytopenic purpura (TTP). It has been observed that cyclosporin A (CsA) treatment, particularly in transplant patients, may sometimes be linked to the development of TTP. Until now, the reason for such a link was unclear. Here we provide evidence demonstrating that cyclophilin B (CypB) activity plays an important role in the secretion of active ADAMTS13. We found that CsA, an inhibitor of CypB, reduces the secretion of ADAMTS13 and leads to conformational changes in the protein resulting in diminished ADAMTS13 proteolytic activity. A direct, functional interaction between CypB (which possesses peptidyl-prolyl cis-trans isomerase (PPIase) and chaperone functions) and ADAMTS13 is demonstrated using immunoprecipitation and siRNA knockdown of CypB. Finally, CypB knock-out mice were found to have reduced ADAMTS13 levels. Taken together, our findings indicate that cyclophilin-mediated activity is an important factor affecting secretion and activity of ADAMTS13. The large number of proline residues in ADAMTS13 is consistent with the important role of cis-trans isomerization in the proper folding of this protein. These results altogether provide a novel mechanistic explanation for CsA-induced TTP in transplant patients. PMID:23144461

  5. MicroRNA-145 Targets the Metalloprotease ADAM17 and Is Suppressed in Renal Cell Carcinoma Patients1

    PubMed Central

    Doberstein, Kai; Steinmeyer, Nico; Hartmetz, Ann-Kathrin; Eberhardt, Wolfgang; Mittelbronn, Michel; Harter, Patrick N; Juengel, Eva; Blaheta, Roman; Pfeilschifter, Josef; Gutwein, Paul

    2013-01-01

    A disintegrin and metalloproteinase 17 (ADAM17) is a metalloprotease that is overexpressed in many cancer types, including renal cancers. However, the regulatory mechanisms of ADAM17 in cancer development and progression are poorly understood. In the present work, we provide evidence using overexpression and inhibition of microRNA 145 (miR-145) that miR-145 negatively regulates ADAM17 expression. Furthermore, we show that ADAM17 negatively regulates miR-145 through tumor necrosis factor-α, resulting in a reciprocal negative feedback loop. In this study, the expression of ADAM17 and miR-145 correlated negatively in renal cancer tumor tissues and cell lines, suggesting an important regulatory mechanism. Additionally, we showed that the regulation of ADAM17 is partly involved in the effects of miR-145 on proliferation and migration, whereas no involvement in chemosensitivity was observed. Importantly, in the healthy kidney, miR-145 was detected in different cell types including tubular cells, which are considered the origin of renal cancer. In renal cancer cell lines, miR-145 expression was strongly suppressed by methylation. In summary, miR-145 is downregulated in renal cancer patients, which leads to the up-regulation of ADAM17 in renal cancer. Importantly, miR-145 and ADAM17 are regulated in a reciprocal negative feedback loop. PMID:23441135

  6. Molecular pathology of brain matrix metalloproteases, claudin5, and aquaporins in forensic autopsy cases with special regard to methamphetamine intoxication.

    PubMed

    Wang, Qi; Ishikawa, Takaki; Michiue, Tomomi; Zhu, Bao-Li; Guan, Da-Wei; Maeda, Hitoshi

    2014-05-01

    Methamphetamine (METH) is a highly addictive drug of abuse and toxic to the brain. Recent studies indicated that besides direct damage to dopamine and 5-HT terminals, neurotoxicity of METH may also result from its ability to modify the structure of blood-brain barrier (BBB). The present study investigated the postmortem brain mRNA and immunohistochemical expressions of matrix metalloproteases (MMPs), claudin5 (CLDN5), and aquaporins (AQPs) in forensic autopsy cases of carbon monoxide (n = 14), METH (n = 21), and phenobarbital (n = 17) intoxication, compared with mechanical asphyxia (n = 15), brain injury (n = 11), non-brain injury (n = 21), and sharp instrument injury (n = 15) cases. Relative mRNA quantification using Taqman real-time PCR assay demonstrated higher expression of AQP4 and MMP9, lower expression of CLDN5 in METH intoxication cases and lower expression of MMP2 in phenobarbital intoxication cases. Immunostaining results showed substantial interindividual variations in each group, showing no evident differences in distribution or intensity among all the causes of death. These findings suggest that METH may increase BBB permeability by altering CLDN5 and MMP9, and the self-protective system maybe activated to eliminate accumulating water from the extracellular space of the brain by up-regulating AQP4. Systematic analysis of gene expressions using real-time PCR may be a useful procedure in forensic death investigation.

  7. The role of SnoN in transforming growth factor beta1-induced expression of metalloprotease-disintegrin ADAM12.

    PubMed

    Solomon, Emilia; Li, Hui; Duhachek Muggy, Sara; Syta, Emilia; Zolkiewska, Anna

    2010-07-16

    Increased expression of metalloprotease-disintegrin ADAM12 is a hallmark of several pathological conditions, including cancer, cardiovascular disease, and certain inflammatory diseases of the central nervous system or the muscoskeletal system. We show that transforming growth factor beta1 (TGFbeta1) is a potent inducer of ADAM12 mRNA and protein in mouse fibroblasts and in mouse and human mammary epithelial cells. Induction of ADAM12 is detected within 2 h of treatment with TGFbeta1, is Smad2/Smad3-dependent, and is a result of derepression of the Adam12 gene. SnoN, a negative regulator of the TGFbeta signaling pathway, is a master regulator of ADAM12 expression in response to TGFbeta1 stimulation. Overexpression of SnoN in NIH3T3 cells reduces the magnitude of ADAM12 induction by TGFbeta1 treatment. Down-regulation of SnoN expression by short hairpin RNA enhances TGFbeta1-induced expression of ADAM12. In a panel of TGFbeta1-responsive cancer cell lines with high expression of SnoN, induction of ADAM12 by TGFbeta1 is significantly impaired, suggesting that the endogenous SnoN plays a role in regulating ADAM12 expression in response to TGFbeta1. Identification of SnoN as a repressor of the ADAM12 gene should contribute to advances in the studies on the role of ADAM12 in tumor progression and in the development of other pathologies.

  8. Insulin-Like Growth Factor-I Increases Laminin, Integrin Subunits and Metalloprotease ADAM12 in Mouse Myoblasts.

    PubMed

    Grzelkowska-Kowalczyk, Katarzyna; Grabiec, Kamil; Tokarska, Justyna; Gajewska, Małgorzata; Błaszczyk, Maciej; Milewska, Marta

    2015-01-01

    The extracellular matrix (ECM) is considered a part of the myogenesis signaling mechanism. we hypothesized that insulin-like growth factor-I (IGF-I) modifies ECM during differentiation of mouse C2C12 cells. The myogenic effect of IGF-I (30 nmol/l) was manifested by increased myogenin and myosin heavy chain (MyHC) levels as well as fusion index (2.6 times over control) on the 3rd day of differentiation. IGF-I markedly augmented laminin, but not fibronectin. Cellular contents of integrin α3, α5 and β1 during 3-day differentiation increased in the presence of IGF-I. Treatment with IGF-I increased the expression of the long form of metalloprotease ADAM12 (100 kDa) in myocytes. In conclusion: i) IGF-I caused an increase of laminin, integrin α3 and β1 in C2C12 myogenic cells that can be secondary to stimulation of myogenesis; ii) IGF-I augmented integrin α5 and ADAM12 levels, suggesting a role of this growth factor in determination of the pool of reserve cells during myogenesis.

  9. A highly conserved, inhibitable astacin metalloprotease from Teladorsagia circumcincta is required for cuticle formation and nematode development☆

    PubMed Central

    Stepek, Gillian; McCormack, Gillian; Winter, Alan D.; Page, Antony P.

    2015-01-01

    Parasitic nematodes cause chronic, debilitating infections in both livestock and humans worldwide, and many have developed multiple resistance to the currently available anthelmintics. The protective collagenous cuticle of these parasites is required for nematode survival and its synthesis has been studied extensively in the free-living nematode, Caenorhabditis elegans. The collagen synthesis pathway is a complex, multi-step process involving numerous key enzymes, including the astacin metalloproteases. Nematode astacinsare crucial for C. elegans development, having specific roles in hatching, moulting and cuticle synthesis. NAS-35 (also called DPY-31) is a homologue of a vertebrate procollagen C-proteinase and performs a central role in cuticle formation of C. elegans as its mutation causes temperature-sensitive lethality and cuticle defects. The characterisation of DPY-31 from the ovine gastrointestinal nematode Teladorsagia circumcincta and its ability to rescue the C. elegans mutant is described. Compounds with a hydroxamate functional group have previously been shown to be potent inhibitors of procollagen C-proteinases and were therefore examined for inhibitory activity against the T. circumcincta enzyme. Phenotypic screening against T. circumcincta, Haemonchus contortus and C. elegans larval stages identified compounds that caused body morphology phenotypes consistent with the inhibition of proteases involved in cuticle collagen synthesis. These compounds correspondingly inhibited the activity of recombinant T. circumcincta DPY-31, supporting the hypothesis that this enzyme may represent a potentially novel anthelmintic drug target. PMID:25736599

  10. Intracellular Cleavage of the Cx43 C-Terminal Domain by Matrix-Metalloproteases: A Novel Contributor to Inflammation?

    PubMed Central

    De Bock, Marijke; Wang, Nan; Decrock, Elke; Bultynck, Geert; Leybaert, Luc

    2015-01-01

    The coordination of tissue function is mediated by gap junctions (GJs) that enable direct cell-cell transfer of metabolic and electric signals. GJs are formed by connexin (Cx) proteins of which Cx43 is most widespread in the human body. Beyond its role in direct intercellular communication, Cx43 also forms nonjunctional hemichannels (HCs) in the plasma membrane that mediate the release of paracrine signaling molecules in the extracellular environment. Both HC and GJ channel function are regulated by protein-protein interactions and posttranslational modifications that predominantly take place in the C-terminal domain of Cx43. Matrix metalloproteases (MMPs) are a major group of zinc-dependent proteases, known to regulate not only extracellular matrix remodeling, but also processing of intracellular proteins. Together with Cx43 channels, both GJs and HCs, MMPs contribute to acute inflammation and a small number of studies reports on an MMP-Cx43 link. Here, we build further on these reports and present a novel hypothesis that describes proteolytic cleavage of the Cx43 C-terminal domain by MMPs and explores possibilities of how such cleavage events may affect Cx43 channel function. Finally, we set out how aberrant channel function resulting from cleavage can contribute to the acute inflammatory response during tissue injury. PMID:26424967

  11. Degradation of circulating von Willebrand factor and its regulator ADAMTS13 implicates secreted Bacillus anthracis metalloproteases in anthrax consumptive coagulopathy.

    PubMed

    Chung, Myung-Chul; Popova, Taissia G; Jorgensen, Shelley C; Dong, Li; Chandhoke, Vikas; Bailey, Charles L; Popov, Serguei G

    2008-04-11

    Pathology data from the anthrax animal models show evidence of significant increases in vascular permeability coincident with hemostatic imbalances manifested by thrombocytopenia, transient leucopenia, and aggressive disseminated intravascular coagulation. In this study we hypothesized that anthrax infection modulates the activity of von Willebrand factor (VWF) and its endogenous regulator ADAMTS13, which play important roles in hemostasis and thrombosis, including interaction of endothelial cells with platelets. We previously demonstrated that purified anthrax neutral metalloproteases Npr599 and InhA are capable of cleaving a variety of host structural and regulatory proteins. Incubation of human plasma with these proteases at 37 degrees C in the presence of urea as a mild denaturant results in proteolysis of VWF. Also in these conditions, InhA directly cleaves plasma ADAMTS13 protein. Npr599 and InhA digest synthetic VWF substrate FRETS-VWF73. Amino acid sequencing of VWF fragments produced by InhA suggests that one of the cleavage sites of VWF is located at domain A2, the target domain of ADAMTS13. Proteolysis of VWF by InhA impairs its collagen binding activity (VWF:CBA) and ristocetin-induced platelet aggregation activity. In plasma from anthrax spore-challenged DBA/2 mice, VWF antigen levels increase up to 2-fold at day 3 post-infection with toxigenic Sterne 34F(2) strain, whereas VWF:CBA levels drop in a time-dependent manner, suggesting dysfunction of VWF instead of its quantitative deficiency. This conclusion is further supported by significant reduction in the amount of VWF circulating in blood in the ultra-large forms. In addition, Western blot analysis shows proteolytic depletion of ADAMTS13 from plasma of spore-challenged mice despite its increased expression in the liver. Our results suggest a new mechanism of anthrax coagulopathy affecting the levels and functional activities of both VWF and its natural regulator ADAMTS13. This mechanism may

  12. YghJ, the secreted metalloprotease of pathogenic E. coli induces hemorrhagic fluid accumulation in mouse ileal loop.

    PubMed

    Tapader, Rima; Bose, Dipro; Pal, Amit

    2017-04-01

    YghJ, also known as SslE (Secreted and surface associated lipoprotein) is a cell surface associated and secreted lipoprotein harbouring M60 metalloprotease domain. Though the gene is known to be conserved among both pathogenic and commensal Escherichia coli isolates, the expression and secretion of YghJ was found to be higher among diverse E. coli pathotypes. YghJ, secreted from intestinal pathogens such as enterotoxigenic E. coli (ETEC) and enteropathogenic E. coli (EPEC) has been demonstrated to possess mucinase activity and hence facilitates colonization of these enteric pathogens to intestinal epithelial cells. Importantly, YghJ is also reported to be secreted from extraintestinal pathogenic E. coli isolates. In our previous study we have shown that YghJ, purified from a neonatal septicemic E. coli isolate could trigger induction of various proinflammatory cytokines in vitro. This led us to investigate the role of YghJ in causing in vivo tissue hemorrhage. In the present study, we validate the earlier in vitro finding and have showed that YghJ can cause extensive tissue damage in mouse ileum and is also able to induce significant fluid accumulation in a dose dependent manner in a mouse ileal loop (MIL) assay. Hence, our present study not only confirms the pathogenic potential of YghJ in sepsis pathophysiology but also indicates the enterotoxic ability of YghJ which makes it an important virulence determinant of intestinal pathogenic E. coli.

  13. A Metalloprotease (MprIII) Involved in the Chitinolytic System of a Marine Bacterium, Alteromonas sp. Strain O-7

    PubMed Central

    Miyamoto, Katsushiro; Nukui, Eiji; Hirose, Mariko; Nagai, Fumi; Sato, Takaji; Inamori, Yoshihiko; Tsujibo, Hiroshi

    2002-01-01

    Alteromonas sp. strain O-7 secretes several proteins in addition to chitinolytic enzymes in response to chitin induction. In this paper, we report that one of these proteins, designated MprIII, is a metalloprotease involved in the chitin degradation system of the strain. The gene encoding MprIII was cloned in Escherichia coli. The open reading frame of mprIII encoded a protein of 1,225 amino acids with a calculated molecular mass of 137,016 Da. Analysis of the deduced amino acid sequence of MprIII revealed that the enzyme consisted of four domains: the signal sequence, the N-terminal proregion, the protease region, and the C-terminal extension. The C-terminal extension (PkdDf) was characterized by four polycystic kidney disease domains and two domains of unknown function. Western and real-time quantitative PCR analyses demonstrated that mprIII was induced in the presence of insoluble polysaccharides, such as chitin and cellulose. Native MprIII was purified to homogeneity from the culture supernatant of Alteromonas sp. strain O-7 and characterized. The molecular mass of mature MprIII was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 115 kDa. The optimum pH and temperature of MprIII were 7.5 and 50°C, respectively, when gelatin was used as a substrate. Pretreatment of native chitin with MprIII significantly promoted chitinase activity. Furthermore, the combination of MprIII and a novel chitin-binding protease (AprIV) remarkably promoted the chitin hydrolysis efficiency of chitinase. PMID:12406750

  14. Ex Vivo and In Vivo Imaging and Biodistribution of Aptamers Targeting the Human Matrix MetalloProtease-9 in Melanomas

    PubMed Central

    Kryza, David; Debordeaux, Frédéric; Azéma, Laurent; Hassan, Aref; Paurelle, Olivier; Schulz, Jürgen; Savona-Baron, Catherine; Charignon, Elsa; Bonazza, Pauline; Taleb, Jacqueline; Fernandez, Philippe; Janier, Marc; Toulmé, Jean Jacques

    2016-01-01

    The human Matrix MetalloProtease-9 (hMMP-9) is overexpressed in tumors where it promotes the release of cancer cells thus contributing to tumor metastasis. We raised aptamers against hMMP-9, which constitutes a validated marker of malignant tumors, in order to design probes for imaging tumors in human beings. A chemically modified RNA aptamer (F3B), fully resistant to nucleases was previously described. This compound was subsequently used for the preparation of F3B-Cy5, F3B-S-acetylmercaptoacetyltriglycine (MAG) and F3B-DOTA. The binding properties of these derivatives were determined by surface plasmon resonance and electrophoretic mobility shift assay. Optical fluorescence imaging confirmed the binding to hMMP-9 in A375 melanoma bearing mice. Quantitative biodistribution studies were performed at 30 min, 1h and 2 h post injection of 99mTc-MAG-aptamer and 111In-DOTA-F3B. 99mTc radiolabeled aptamer specifically detected hMMP-9 in A375 melanoma tumors but accumulation in digestive tract was very high. Following i.v. injection of 111In-DOTA-F3B, high level of radioactivity was observed in kidneys and bladder but digestive tract uptake was very limited. Tumor uptake was significantly (student t test, p<0.05) higher for 111In-DOTA-F3B with 2.0%ID/g than for the 111In-DOTA-control oligonucleotide (0.7%ID/g) with tumor to muscle ratio of 4.0. Such difference in tumor accumulation has been confirmed by ex vivo scintigraphic images performed at 1h post injection and by autoradiography, which revealed the overexpression of hMMP-9 in sections of human melanomas. These results demonstrate that F3B aptamer is of interest for detecting hMMP-9 in melanoma tumor. PMID:26901393

  15. A Model for the Conformational Activation of the Structurally Quiescent Metalloprotease ADAMTS13 by Von Willebrand Factor.

    PubMed

    South, Kieron; Freitas, Marta O; Lane, David A

    2017-02-16

    Blood loss is prevented by the multi-domain glycoprotein von Willebrand factor (VWF), which binds exposed collagen at damaged vessels and captures platelets. VWF is regulated by the metalloprotease ADAMTS13, which, in turn, is conformationally activated by VWF. To delineate the structural requirements for VWF-mediated conformational activation of ADAMTS13, we performed binding and functional studies with a panel of truncated ADAMTS13 variants. We demonstrate that both the isolated CUB1 and CUB2 domains in ADAMTS13 bind to the spacer domain exosite of a truncated ADAMTS13 variant, MDTCS (KD of 135 ± 10.1 nM and 86.9 ± 9.0 nM, respectively). However, only the CUB1 domain inhibited proteolytic activity of MDTCS. Moreover, ADAMTS13∆CUB2, unlike ADAMTS13∆CUB1-2, exhibited activity similar to wild-type ADAMTS13 and could be activated by VWF D4-CK. The CUB2 domain is therefore not essential for maintaining the inactive conformation of ADAMTS13. Both CUB domains could bind to the VWF D4-CK domain fragment (KD of 53.7± 2.1 nM and 84.3 ± 2.0 nM, respectively). However, deletion of both CUB domains did not prevent VWF D4-CK binding, suggesting that competition for CUB-domain binding to the spacer domain is not the dominant mechanism behind the conformational activation. ADAMTS13∆TSP8-CUB2 could no longer bind to VWF D4-CK, and deletion of TSP8 abrogated ADAMTS13 conformational activation. These findings support an ADAMTS13-activation model in which VWF D4-CK engages the TSP8-CUB2 domains, inducing the conformational change that disrupts the CUB1-spacer domain interaction and thereby activates ADAMTS13.

  16. The disintegrin/metalloprotease ADAM 10 is essential for Notch signalling but not for alpha-secretase activity in fibroblasts.

    PubMed

    Hartmann, Dieter; de Strooper, Bart; Serneels, Lutgarde; Craessaerts, Katleen; Herreman, An; Annaert, Wim; Umans, Lieve; Lübke, Torben; Lena Illert, Anna; von Figura, Kurt; Saftig, Paul

    2002-10-01

    The metalloprotease ADAM 10 is an important APP alpha-secretase candidate, but in vivo proof of this is lacking. Furthermore, invertebrate models point towards a key role of the ADAM 10 orthologues Kuzbanian and sup-17 in Notch signalling. In the mouse, this function is, however, currently attributed to ADAM 17/TACE, while the role of ADAM 10 remains unknown. We have created ADAM 10-deficient mice. They die at day 9.5 of embryogenesis with multiple defects of the developing central nervous system, somites, and cardiovascular system. In situ hybridization revealed a reduced expression of the Notch target gene hes-5 in the neural tube and an increased expression of the Notch ligand dll-1, supporting an important role for ADAM 10 in Notch signalling in the vertebrates as well. Since the early lethality precluded the establishment of primary neuronal cultures, APPs alpha generation was analyzed in embryonic fibroblasts and found to be preserved in 15 out of 17 independently generated ADAM 10-deficient fibroblast cell lines, albeit at a quantitatively more variable level than in controls, whereas a severe reduction was found in only two cases. The variability was not due to differences in genetic background or to variable expression of the alternative alpha-secretase candidates ADAM 9 and ADAM 17. These results indicate, therefore, either a regulation between ADAMs on the post-translational level or that other, not yet known, proteases are able to compensate for ADAM 10 deficiency. Thus, the observed variability, together with recent reports on tissue-specific expression patterns of ADAMs 9, 10 and 17, points to the existence of tissue-specific 'teams' of different proteases exerting alpha-secretase activity.

  17. Wild-type amyloid beta 1-40 peptide induces vascular smooth muscle cell death independently from matrix metalloprotease activity.

    PubMed

    Blaise, Régis; Mateo, Véronique; Rouxel, Clotilde; Zaccarini, François; Glorian, Martine; Béréziat, Gilbert; Golubkov, Vladislav S; Limon, Isabelle

    2012-06-01

    Cerebral amyloid angiopathy (CAA) is an important cause of intracerebral hemorrhages in the elderly, characterized by amyloid-β (Aβ) peptide accumulating in central nervous system blood vessels. Within the vessel walls, Aβ-peptide deposits [composed mainly of wild-type (WT) Aβ(1-40) peptide in sporadic forms] induce impaired adhesion of vascular smooth muscle cells (VSMCs) to the extracellular matrix (ECM) associated with their degeneration. This process often results in a loss of blood vessel wall integrity and ultimately translates into cerebral ischemia and microhemorrhages, both clinical features of CAA. In this study, we decipher the molecular mechanism of matrix metalloprotease (MMP)-2 activation in WT-Aβ(1-40) -treated VSMC and provide evidence that MMP activity, although playing a critical role in cell detachment disrupting ECM components, is not involved in the WT-Aβ(1-40) -induced degeneration of VSMCs. Indeed, whereas this peptide clearly induced VSMC apoptosis, neither preventing MMP-2 activity nor hampering the expression of membrane type1-MMP, or preventing tissue inhibitors of MMPs-2 (TIMP-2) recruitment (two proteins evidenced here as involved in MMP-2 activation), reduced the number of dead cells. Even the use of broad-range MMP inhibitors (GM6001 and Batimastat) did not affect WT-Aβ(1-40) -induced cell apoptosis. Our results, in contrast to those obtained using the Aβ(1-40) Dutch variant suggesting a link between MMP-2 activity, VSMC mortality and degradation of specific matrix components, indicate that the ontogenesis of the Dutch familial and sporadic forms of CAAs is different. ECM degradation and VSMC degeneration would be tightly connected in the Dutch familial form while being two independent processes in sporadic forms of CAA.

  18. The Disintegrin and Metalloprotease ADAM12 Is Associated with TGF-β-Induced Epithelial to Mesenchymal Transition.

    PubMed

    Ruff, Michaël; Leyme, Anthony; Le Cann, Fabienne; Bonnier, Dominique; Le Seyec, Jacques; Chesnel, Franck; Fattet, Laurent; Rimokh, Ruth; Baffet, Georges; Théret, Nathalie

    2015-01-01

    The increased expression of the Disintegrin and Metalloprotease ADAM12 has been associated with human cancers, however its role remain unclear. We have previously reported that ADAM12 expression is induced by the transforming growth factor, TGF-β and promotes TGF-β-dependent signaling through interaction with the type II receptor of TGF-β. Here we explore the implication of ADAM12 in TGF-β-mediated epithelial to mesenchymal transition (EMT), a key process in cancer progression. We show that ADAM12 expression is correlated with EMT markers in human breast cancer cell lines and biopsies. Using a non-malignant breast epithelial cell line (MCF10A), we demonstrate that TGF-β-induced EMT increases expression of the membrane-anchored ADAM12L long form. Importantly, ADAM12L overexpression in MCF10A is sufficient to induce loss of cell-cell contact, reorganization of actin cytoskeleton, up-regulation of EMT markers and chemoresistance. These effects are independent of the proteolytic activity but require the cytoplasmic tail and are specific of ADAM12L since overexpression of ADAM12S failed to induce similar changes. We further demonstrate that ADAM12L-dependent EMT is associated with increased phosphorylation of Smad3, Akt and ERK proteins. Conversely, inhibition of TGF-β receptors or ERK activities reverses ADAM12L-induced mesenchymal phenotype. Together our data demonstrate that ADAM12L is associated with EMT and contributes to TGF-β-dependent EMT by favoring both Smad-dependent and Smad-independent pathways.

  19. Expression of metalloprotease insulin-degrading enzyme (insulysin) in normal and malignant human tissues

    PubMed Central

    Yfanti, Christina; Mengele, Karin; Gkazepis, Apostolos; Weirich, Gregor; Giersig, Cecylia; Kuo, Wen-Liang; Tang, Wei-Jen; Rosner, Marsha; Schmitt, Manfred

    2013-01-01

    Background Insulin-degrading enzyme (IDE, insulysin, insulinase; EC 3.4.22.11), a thiol metalloendopeptidase, is involved in intracellular degradation of insulin, thereby inhibiting its translocation and accumulation to the nucleus. Recently, protein expression of IDE has been demonstrated in the epithelial ducts of normal breast and in breast cancer tissue (Radulescu et al., Int J Oncol 30:73; 2007). Materials and Methods Utilizing four different antibodies generated against different epitopes of the IDE molecule, we performed western blot analysis and immunohistochemical staining on several normal human tissues, on a plethora of tumor cell lines of different tissue origin, and on malignant breast and ovarian tissue. Results Applying the four IDE-directed antibodies, we demonstrate IDE expression at the protein level, both by means of immunoblotting and immunocytochemistry, in all of the tumor cell lines analyzed. Besides, IDE protein expression was found in normal tissues of the kidney, liver, lung, brain, breast and skeletal muscle, as well as in breast and ovarian cancer tissues. Immunohistochemical visualization of IDE indicated cytoplasmic localization of IDE in all of the cell lines and tissues assessed. Conclusions We performed for the first time a wide-ranging survey on IDE protein expression in normal and malignant tissues and cells and thus extend knowledge about cellular and tissue distribution of IDE, an enzyme which so far has mainly been studied in connection with Alzheimer’s disease and diabetes but not in cancer. PMID:18813847

  20. Molecular cloning and characterisation of in vitro immune response against astacin-like metalloprotease Ace-MTP-2 from Ancylostoma ceylanicum.

    PubMed

    Bąska, Piotr; Wiśniewski, Marcin; Krzyżowska, Małgorzata; Długosz, Ewa; Zygner, Wojciech; Górski, Paweł; Wędrychowicz, Halina

    2013-04-01

    Ancylostoma ceylanicum belongs to the group of parasites commonly known as hookworms, blood-sucking nematodes which infect around 576 million people and hundreds of millions of animals. The interactions between these parasites and host immune systems are complicated and yet to be determined. Hookworm infections are usually long lasting and recurrent, due in part to their ability to synthesize macromolecules capable of modulating the host immune response. The interaction of parasite proteins with host immune systems has been proven, but so far there is no data describing the influence of astacin-like metalloproteases (expressed among different parasitic nematodes) on the human immune system. The cDNA encoding A. ceylanicum metalloprotease 2 (Ace-mtp-2) was cloned using RACE-PCR. Computational analysis was used to examine the immunogenicity and recombinant Ace-MTP-2 was used to investigate its influence on human THP-1 monocytes and macrophages. The Ace-mtp-2 gene encodes an astascin-like metalloprotease, with a theoretical molecular mass of 26.7 kDa. The protease has a putative signal peptide, 11 potential phosphorylation sites, and two disulfide bridges revealed by computational analysis. Maximal expression of Ace-mtp-2 by A. ceylanicum occurs in the adult stage of the parasite, and Western blot indicates the secretory nature of the protease. This suggests the protease is working at the host-parasite interface and would likely be exposed to the hosts immune response. Recombinant protein were expressed in Escherichia coli and Pichia pastoris. Recombinant Ace-MTP-2 amplified the in vitro release of TNFα and induced release of IFNγ by lipopolysaccharide activated THP-1 macrophages. The presence of Ace-MTP-2 in secretory products of the adult parasite and the induction of IFNγ release may suggest an important role for Ace-MTP-2 in host-parasite interactions since IFNγ is suggested to be responsible for the protective immune response against adult hookworms.

  1. Glycosyl-Phosphatidyl-Inositol (GPI)-Anchors and Metalloproteases: Their Roles in the Regulation of Exosome Composition and NKG2D-Mediated Immune Recognition.

    PubMed

    López-Cobo, Sheila; Campos-Silva, Carmen; Valés-Gómez, Mar

    2016-01-01

    Communication within the immune system depends on the release of factors that can travel and transmit information at points distant from the cell that produced them. In general, immune cells use two key strategies that can occur either at the plasma membrane or in intracellular compartments to produce such factors, vesicle release and proteolytic cleavage. Release of soluble factors in exosomes, a subset of vesicles that originate from intracellular compartments, depends generally on biochemical and lipid environment features. This physical environment allows proteins to be recruited to membrane microdomains that will be later endocytosed and further released to the extracellular milieu. Cholesterol and sphingolipid rich domains (also known as lipid rafts or detergent-resistant membranes, DRMs) often contribute to exosomes and these membrane regions are rich in proteins modified with Glycosyl-Phosphatidyl-Inositol (GPI) and lipids. For this reason, many palmitoylated and GPI-anchored proteins are preferentially recruited to exosomes. In this review, we analyse the biochemical features involved in the release of NKG2D-ligands as an example of functionally related gene families encoding both transmembrane and GPI-anchored proteins that can be released either by proteolysis or in exosomes, and modulate the intensity of the immune response. The immune receptor NKG2D is present in all human Natural Killer and T cells and plays an important role in the first barrier of defense against tumor and infection. However, tumor cells can evade the immune system by releasing NKG2D-ligands to induce down-regulation of the receptor. Some NKG2D-ligands can be recruited to exosomes and potently modulate receptor expression and immune function, while others are more susceptible to metalloprotease cleavage and are shed as soluble molecules. Strikingly, metalloprotease inhibition is sufficient to drive the accumulation in exosomes of ligands otherwise released by metalloprotease

  2. Glycosyl-Phosphatidyl-Inositol (GPI)-Anchors and Metalloproteases: Their Roles in the Regulation of Exosome Composition and NKG2D-Mediated Immune Recognition

    PubMed Central

    López-Cobo, Sheila; Campos-Silva, Carmen; Valés-Gómez, Mar

    2016-01-01

    Communication within the immune system depends on the release of factors that can travel and transmit information at points distant from the cell that produced them. In general, immune cells use two key strategies that can occur either at the plasma membrane or in intracellular compartments to produce such factors, vesicle release and proteolytic cleavage. Release of soluble factors in exosomes, a subset of vesicles that originate from intracellular compartments, depends generally on biochemical and lipid environment features. This physical environment allows proteins to be recruited to membrane microdomains that will be later endocytosed and further released to the extracellular milieu. Cholesterol and sphingolipid rich domains (also known as lipid rafts or detergent-resistant membranes, DRMs) often contribute to exosomes and these membrane regions are rich in proteins modified with Glycosyl-Phosphatidyl-Inositol (GPI) and lipids. For this reason, many palmitoylated and GPI-anchored proteins are preferentially recruited to exosomes. In this review, we analyse the biochemical features involved in the release of NKG2D-ligands as an example of functionally related gene families encoding both transmembrane and GPI-anchored proteins that can be released either by proteolysis or in exosomes, and modulate the intensity of the immune response. The immune receptor NKG2D is present in all human Natural Killer and T cells and plays an important role in the first barrier of defense against tumor and infection. However, tumor cells can evade the immune system by releasing NKG2D-ligands to induce down-regulation of the receptor. Some NKG2D-ligands can be recruited to exosomes and potently modulate receptor expression and immune function, while others are more susceptible to metalloprotease cleavage and are shed as soluble molecules. Strikingly, metalloprotease inhibition is sufficient to drive the accumulation in exosomes of ligands otherwise released by metalloprotease

  3. Doxycycline, a matrix metalloprotease inhibitor, reduces vascular remodeling and damage after cerebral ischemia in stroke-prone spontaneously hypertensive rats.

    PubMed

    Pires, Paulo W; Rogers, Curt T; McClain, Jonathon L; Garver, Hannah S; Fink, Gregory D; Dorrance, Anne M

    2011-07-01

    Matrix metalloproteases (MMPs) are a family of zinc peptidases involved in extracellular matrix turnover. There is evidence that increased MMP activity is involved in remodeling of resistance vessels in chronic hypertension. Thus we hypothesized that inhibition of MMP activity with doxycycline (DOX) would attenuate vascular remodeling. Six-week-old male stroke-prone spontaneously hypertensive rats (SHRSP) were treated with DOX (50 mg·kg(-1)·day(-1) in the drinking water) for 6 wk. Untreated SHRSP were controls. Blood pressure was measured by telemetry during the last week. Middle cerebral artery (MCA) and mesenteric resistance artery (MRA) passive structures were assessed by pressure myography. MMP-2 expression in aortas was measured by Western blot. All results are means ± SE. DOX caused a small increase in mean arterial pressure (SHRSP, 154 ± 1; SHRSP + DOX, 159 ± 3 mmHg; P < 0.001). Active MMP-2 expression was reduced in aorta from SHRSP + DOX (0.21 ± 0.06 vs. 0.49 ± 0.13 arbitrary units; P < 0.05). In the MCA, at 80 mmHg, DOX treatment increased the lumen (273.2 ± 4.7 vs. 238.3 ± 6.3 μm; P < 0.05) and the outer diameter (321 ± 5.3 vs. 290 ± 7.6 μm; P < 0.05) and reduced the wall-to-lumen ratio (0.09 ± 0.002 vs. 0.11 ± 0.003; P < 0.05). Damage after transient cerebral ischemia (transient MCA occlusion) was reduced in SHRSP + DOX (20.7 ± 4 vs. 45.5 ± 5% of hemisphere infarcted; P < 0.05). In the MRA, at 90 mmHg DOX, reduced wall thickness (29 ± 1 vs. 22 ± 1 μm; P < 0.001) and wall-to-lumen ratio (0.08 ± 0.004 vs. 0.11 ± 0.008; P < 0.05) without changing lumen diameter. These results suggest that MMPs are involved in hypertensive vascular remodeling in both the peripheral and cerebral vasculature and that DOX reduced brain damage after cerebral ischemia.

  4. MALDI-TOF MS and CD Spectral Analysis for Identification and Structure Prediction of a Purified, Novel, Organic Solvent Stable, Fibrinolytic Metalloprotease from Bacillus cereus B80

    PubMed Central

    Saxena, Rajshree

    2015-01-01

    The ability to predict protein function from structure is becoming increasingly important; hence, elucidation and determination of protein structure become the major steps in proteomics. The present study was undertaken for identification of metalloprotease produced by Bacillus cereus B80 and recognition of characteristics that can be industrially exploited. The enzyme was purified in three steps combining precipitation and chromatographic methods resulting in 33.5% recovery with 13.1-fold purification of enzyme which was detected as a single band with a molecular mass of 26 kDa approximately in SDS-PAGE and zymogram. The MALDI-TOF MS showed that the enzyme exhibited 70–93% similarity with zinc metalloproteases from various strains Bacillus sp. specifically from Bacillus cereus group. The sequence alignment revealed the presence of zinc-binding region VVVHEMCHMV in the most conserved C terminus region. Secondary structure of the enzyme was obtained by CD spectra and I-TASSER. The enzyme kinetics revealed a Michaelis constant (Km) of 0.140 μmol/ml and Vmax of 2.11 μmol/min. The application studies showed that the enzyme was able to hydrolyze various proteins with highest affinity towards casein followed by BSA and gelatin. The enzyme exhibited strong fibrinolytic, collagenolytic, and gelatinolytic properties and stability in various organic solvents. PMID:25802851

  5. Aminopeptidase N1 (EtAPN1), an M1 metalloprotease of the apicomplexan parasite Eimeria tenella, participates in parasite development.

    PubMed

    Gras, Simon; Byzia, Anna; Gilbert, Florence B; McGowan, Sheena; Drag, Marcin; Silvestre, Anne; Niepceron, Alisson; Lecaille, Fabien; Lalmanach, Gilles; Brossier, Fabien

    2014-07-01

    Aminopeptidases N are metalloproteases of the M1 family that have been reported in numerous apicomplexan parasites, including Plasmodium, Toxoplasma, Cryptosporidium, and Eimeria. While investigating the potency of aminopeptidases as therapeutic targets against coccidiosis, one of the most important avian diseases caused by the genus Eimeria, we identified and characterized Eimeria tenella aminopeptidase N1 (EtAPN1). Its inhibition by bestatin and amastatin, as well as its reactivation by divalent ions, is typical of zinc-dependent metalloproteases. EtAPN1 shared a similar sequence, three-dimensional structure, and substrate specificity and similar kinetic parameters with A-M1 from Plasmodium falciparum (PfA-M1), a validated target in the treatment of malaria. EtAPN1 is synthesized as a 120-kDa precursor and cleaved into 96-, 68-, and 38-kDa forms during sporulation. Further, immunolocalization assays revealed that, similar to PfA-M1, EtAPN1 is present during the intracellular life cycle stages in both the parasite cytoplasm and the parasite nucleus. The present results support the hypothesis of a conserved role between the two aminopeptidases, and we suggest that EtAPN1 might be a valuable target for anticoccidiosis drugs.

  6. A Natural Bacterial-Derived Product, the Metalloprotease Arazyme, Inhibits Metastatic Murine Melanoma by Inducing MMP-8 Cross-Reactive Antibodies

    PubMed Central

    Pereira, Felipe V.; Ferreira-Guimarães, Carla A.; Paschoalin, Thaysa; Scutti, Jorge A. B.; Melo, Filipe M.; Silva, Luis S.; Melo, Amanda C. L.; Silva, Priscila; Tiago, Manoela; Matsuo, Alisson L.; Juliano, Luiz; Juliano, Maria A.; Carmona, Adriana K.; Travassos, Luiz R.; Rodrigues, Elaine G.

    2014-01-01

    The increased incidence, high rates of mortality and few effective means of treatment of malignant melanoma, stimulate the search for new anti-tumor agents and therapeutic targets to control this deadly metastatic disease. In the present work the antitumor effect of arazyme, a natural bacterial-derived metalloprotease secreted by Serratia proteomaculans, was investigated. Arazyme significantly reduced the number of pulmonary metastatic nodules after intravenous inoculation of B16F10 melanoma cells in syngeneic mice. In vitro, the enzyme showed a dose-dependent cytostatic effect in human and murine tumor cells, and this effect was associated to the proteolytic activity of arazyme, reducing the CD44 expression at the cell surface, and also reducing in vitro adhesion and in vitro/in vivo invasion of these cells. Arazyme treatment or immunization induced the production of protease-specific IgG that cross-reacted with melanoma MMP-8. In vitro, this antibody was cytotoxic to tumor cells, an effect increased by complement. In vivo, arazyme-specific IgG inhibited melanoma lung metastasis. We suggest that the antitumor activity of arazyme in a preclinical model may be due to a direct cytostatic activity of the protease in combination with the elicited anti-protease antibody, which cross-reacts with MMP-8 produced by tumor cells. Our results show that the bacterial metalloprotease arazyme is a promising novel antitumor chemotherapeutic agent. PMID:24788523

  7. Expression of tissue inhibitor of metalloprotease 3 is reduced in ischemic but not neuropathic ulcers from patients with type 2 diabetes mellitus.

    PubMed

    Menghini, R; Uccioli, L; Vainieri, E; Pecchioli, C; Casagrande, V; Stoehr, R; Cardellini, M; Porzio, O; Rizza, S; Federici, M

    2013-12-01

    Diabetic foot ulceration remains one of the most common and most serious consequences of diabetes. Persistently high levels of matrix metalloproteases (MMPs) contribute to wound chronicity. Our aim was to assess the concentrations of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in neuropathic and ischemic diabetic foot ulcers by analyzing biopsy samples. In this study, biopsies were taken from 35 diabetic foot ulcers of type 2 diabetes mellitus patients and distinguished in neuropathic (n = 14) or ischemic (n = 21). Zymography assay was utilized for the analysis of MMP-2 and MMP-9 activity. TACE activity was evaluated by a specific fluorimetric assay. mRNA levels of MMPs as well as TIMPs were detected using quantitative real-time polymerase chain reaction. The activity of MMP9 and A Disintegrin and A MetalloProtease Domain 17/TNF-Alpha Converting Enzyme (ADAM17/TACE) was significantly increased in ischemic compared to neuropathic biopsies. No differences were detected between both groups for the mRNA levels of MMPs as well as of ADAMs. However, TIMP3 mRNA expression was decreased in ischemic samples. The combination of increased activity of MMP9 and ADAM17/TACE with decreased concentrations of TIMP-3 mRNA expression in ischemic diabetic foot ulcers compared to neuropathic samples suggests that the increased proteolytic environment may represent a causative factor in the ulcer progression. New treatment strategies for healing diabetic foot ulcers could be directed toward increasing levels of TIMP3.

  8. Precipitation of Iron on the Surface of Leptospira interrogans Is Associated with Mutation of the Stress Response Metalloprotease HtpX

    PubMed Central

    Henry, Rebekah; Lo, Miranda; Khoo, Chenai; Zhang, Hailong; Boysen, Reinhard I.; Picardeau, Mathieu; Murray, Gerald L.; Bulach, Dieter M.

    2013-01-01

    High concentrations of free metal ions in the environment can be detrimental to bacterial survival. However, bacteria utilize strategies, including the activation of stress response pathways and immobilizing chemical elements on their surface, to limit this toxicity. In this study, we characterized LA4131, the HtpX-like M48 metalloprotease from Leptospira interrogans, with a putative role in bacterial stress response and membrane homeostasis. Growth of the la4131 transposon mutant strain (L522) in 360 μM FeSO4 (10-fold the normal in vitro concentration) resulted in the production of an amorphous iron precipitate. Atomic force microscopy and transmission electron microscopy analysis of the strain demonstrated that precipitate production was associated with the generation and release of outer membrane vesicles (OMVs) from the leptospiral surface. Transcriptional studies indicated that inactivation of la4131 resulted in altered expression of a subset of metal toxicity and stress response genes. Combining these findings, this report describes OMV production in response to environmental stressors and associates OMV production with the in vitro activity of an HtpX-like metalloprotease. PMID:23709510

  9. H-ras oncogene-transformed human bronchial epithelial cells (TBE-1) secrete a single metalloprotease capable of degrading basement membrane collagen

    SciTech Connect

    Collier, I.E.; Wilhelm, S.M.; Eisen, A.Z.; Marmer, B.L.; Grant, G.A.; Seltzer, J.L.; Kronberger, A.; He, C.; Bauer, E.A.; Goldberg, G.I.

    1988-05-15

    H-ras transformed human bronchial epithelial cells (TBE-1) secrete a single major extracellular matrix metalloprotease which is not found in the normal parental cells. The enzyme is secreted in a latent form which can be activated to catalyze the cleavage of the basement membrane macromolecule type IV collagen. The substrates in their order of preference are: gelatin, type IV collagen, type V collagen, fibronectin, and type VII collagen; but the enzyme does not cleave the interstitial collagens or laminin. This protease is identical to gelatinase isolated from normal human skin explants, normal human skin fibroblasts, and SV40-transformed human lung fibroblasts. Based on this ability to initiate the degradation of type IV collagen in a pepsin-resistant portion of the molecule, it will be referred to as type IV collagenase. This enzyme is most likely the human analog of type IV collagenase detected in several rodent tumors. Type IV collagenase consists of three domains. Type IV collagenase represents the third member of a newly recognized gene family coding for secreted extracellular matrix metalloproteases, which includes interstitial fibroblast collagenase and stromelysin.

  10. A smallest 6 kda metalloprotease, mini-matrilysin, in living world: a revolutionary conserved zinc-dependent proteolytic domain- helix-loop-helix catalytic zinc binding domain (ZBD)

    PubMed Central

    2012-01-01

    Background The Aim of this study is to study the minimum zinc dependent metalloprotease catalytic folding motif, helix B Met loop-helix C, with proteolytic catalytic activities in metzincin super family. The metzincin super family share a catalytic domain consisting of a twisted five-stranded β sheet and three long α helices (A, B and C). The catalytic zinc is at the bottom of the cleft and is ligated by three His residues in the consensus sequence motif, HEXXHXXGXXH, which is located in helix B and part of the adjacent Met turn region. An interesting question is - what is the minimum portion of the enzyme that still possesses catalytic and inhibitor recognition?” Methods We have expressed a 60-residue truncated form of matrilysin which retains only the helix B-Met turn-helix C region and deletes helix A and the five-stranded β sheet which form the upper portion of the active cleft. This is only 1/4 of the full catalytic domain. The E. coli derived 6 kDa MMP-7 ZBD fragments were purified and refolded. The proteolytic activities were analyzed by Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 peptide assay and CM-transferrin zymography analysis. SC44463, BB94 and Phosphoramidon were computationally docked into the 3day structure of the human MMP7 ZBD and TAD and thermolysin using the docking program GOLD. Results This minimal 6 kDa matrilysin has been refolded and shown to have proteolytic activity in the Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 peptide assay. Triton X-100 and heparin are important factors in the refolding environment for this mini-enzyme matrilysin. This minienzyme has the proteolytic activity towards peptide substrate, but the hexamer and octamer of the mini MMP-7 complex demonstrates the CM-transferrin proteolytic activities in zymographic analysis. Peptide digestion is inhibited by SC44463, specific MMP7 inhibitors, but not phosphorimadon. Interestingly, the mini MMP-7 can be processed by autolysis and producing ~ 6 ~ 7 kDa fragments. Thus

  11. Moojenactivase, a novel pro-coagulant PIIId metalloprotease isolated from Bothrops moojeni snake venom, activates coagulation factors II and X and induces tissue factor up-regulation in leukocytes.

    PubMed

    Sartim, Marco A; Costa, Tassia R; Laure, Helen J; Espíndola, Milena S; Frantz, Fabiani G; Sorgi, Carlos A; Cintra, Adélia C O; Arantes, Eliane C; Faccioli, Lucia H; Rosa, José C; Sampaio, Suely V

    2016-05-01

    Coagulopathies following snakebite are triggered by pro-coagulant venom toxins, in which metalloproteases play a major role in envenomation-induced coagulation disorders by acting on coagulation cascade, platelet function and fibrinolysis. Considering this relevance, here we describe the isolation and biochemical characterization of moojenactivase (MooA), a metalloprotease from Bothrops moojeni snake venom, and investigate its involvement in hemostasis in vitro. MooA is a glycoprotein of 85,746.22 Da, member of the PIIId group of snake venom metalloproteases, composed of three linked disulfide-bonded chains: an N-glycosylated heavy chain, and two light chains. The venom protease induced human plasma clotting in vitro by activating on both blood coagulation factors II (prothrombin) and X, which in turn generated α-thrombin and factor Xa, respectively. Additionally, MooA induced expression of tissue factor (TF) on the membrane surface of peripheral blood mononuclear cells (PBMC), which led these cells to adopt pro-coagulant characteristics. MooA was also shown to be involved with production of the inflammatory mediators TNF-α, IL-8 and MCP-1, suggesting an association between MooA pro-inflammatory stimulation of PBMC and TF up-regulation. We also observed aggregation of washed platelets when in presence of MooA; however, the protease had no effect on fibrinolysis. Our findings show that MooA is a novel hemostatically active metalloprotease, which may lead to the development of coagulopathies during B. moojeni envenomation. Moreover, the metalloprotease may contribute to the development of new diagnostic tools and pharmacological approaches applied to hemostatic disorders.

  12. Identification of two components of the Serratia marcescens metalloprotease transporter: protease SM secretion in Escherichia coli is TolC dependent.

    PubMed

    Létoffé, S; Ghigo, J M; Wandersman, C

    1993-11-01

    The Serratia marcescens metalloprotease (protease SM) belongs to a family of proteins secreted from gram-negative bacteria by a signal peptide-independent pathway which requires a specific transporter consisting of three proteins: two in the inner membrane and one in the outer membrane. The prtDSM and prtESM genes encoding the two S. marcescens inner membrane components were cloned and expressed in Escherichia coli. Their nucleotide sequence revealed high overall homology with the two analogous inner membrane components of the Erwinia chrysanthemi protease secretion apparatus and lower, but still significant, homology with the two analogous inner membrane components of the E. coli hemolysin transporter. When expressed in E. coli, these two proteins, PrtDSM and PrtESM, allowed the secretion of protease SM only in the presence of TolC protein, the outer membrane component of the hemolysin transporter.

  13. EphrinA/EphA-induced ectodomain shedding of neural cell adhesion molecule regulates growth cone repulsion through ADAM10 metalloprotease.

    PubMed

    Brennaman, Leann H; Moss, Marcia L; Maness, Patricia F

    2014-01-01

    EphrinA/EphA-dependent axon repulsion is crucial for synaptic targeting in developing neurons but downstream molecular mechanisms remain obscure. Here, it is shown that ephrinA5/EphA3 triggers proteolysis of the neural cell adhesion molecule (NCAM) by the metalloprotease a disintegrin and metalloprotease (ADAM)10 to promote growth cone collapse in neurons from mouse neocortex. EphrinA5 induced ADAM10 activity to promote ectodomain shedding of polysialic acid-NCAM in cortical neuron cultures, releasing a ~ 250 kDa soluble fragment consisting of most of its extracellular region. NCAM shedding was dependent on ADAM10 and EphA3 kinase activity as shown in HEK293T cells transfected with dominant negative ADAM10 and kinase-inactive EphA3 (K653R) mutants. Purified ADAM10 cleaved NCAM at a sequence within the E-F loop of the second fibronectin type III domain (Leu(671) -Lys(672) /Ser(673) -Leu(674) ) identified by mass spectrometry. Mutations of NCAM within the ADAM10 cleavage sequence prevented EphA3-induced shedding of NCAM in HEK293T cells. EphrinA5-induced growth cone collapse was dependent on ADAM10 activity, was inhibited in cortical cultures from NCAM null mice, and was rescued by WT but not ADAM10 cleavage site mutants of NCAM. Regulated proteolysis of NCAM through the ephrin5/EphA3/ADAM10 mechanism likely impacts synapse development, and may lead to excess NCAM shedding when disrupted, as implicated in neurodevelopmental disorders such as schizophrenia. PSA-NCAM and ephrinA/EphA3 coordinately regulate inhibitory synapse development. Here, we have found that ephrinA5 stimulates EphA3 kinase and ADAM10 activity to promote PSA-NCAM cleavage at a site in its second FNIII repeat, which regulates ephrinA5-induced growth cone collapse in GABAergic and non-GABAergic neurons. These findings identify a new regulatory mechanism which may contribute to inhibitory connectivity.

  14. Interaction between TNF and BmooMP-Alpha-I, a Zinc Metalloprotease Derived from Bothrops moojeni Snake Venom, Promotes Direct Proteolysis of This Cytokine: Molecular Modeling and Docking at a Glance

    PubMed Central

    Silva, Maraisa Cristina; Lopes Silva, Tamires; Silva, Murilo Vieira; Mota, Caroline Martins; Santiago, Fernanda Maria; Fonseca, Kelly Cortes; Oliveira, Fábio; Mineo, Tiago Wilson Patriarca; Mineo, José Roberto

    2016-01-01

    Tumor necrosis factor (TNF) is a major cytokine in inflammatory processes and its deregulation plays a pivotal role in several diseases. Here, we report that a zinc metalloprotease extracted from Bothrops moojeni venom (BmooMP-alpha-I) inhibits TNF directly by promoting its degradation. This inhibition was demonstrated by both in vitro and in vivo assays, using known TLR ligands. These findings are supported by molecular docking results, which reveal interaction between BmooMP-alpha-I and TNF. The major cluster of interaction between BmooMP-alpha-I and TNF was confirmed by the structural alignment presenting Ligand Root Mean Square Deviation LRMS = 1.05 Å and Interactive Root Mean Square Deviation IRMS = 1.01 Å, this result being compatible with an accurate complex. Additionally, we demonstrated that the effect of this metalloprotease on TNF is independent of cell cytotoxicity and it does not affect other TLR-triggered cytokines, such as IL-12. Together, these results indicate that this zinc metalloprotease is a potential tool to be further investigated for the treatment of inflammatory disorders involving TNF deregulation. PMID:27447669

  15. Expression of mRNAs coding for VAP1/crotastatin-like metalloproteases in the venom glands of three South American pit vipers assessed by quantitative real-time PCR.

    PubMed

    Tavares, N A C; Correia, J M; Guarnieri, M C; Lima-Filho, J L; Prieto-da-Silva, A R B; Rádis-Baptista, G

    2008-12-15

    Snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. From this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like VAP1 and 2 from the venom of Crotalus atrox, are known. Since most VAP1-like toxins ever characterized were purified from the venom of Viperidae species inhabiting diverse places on earth, we investigate the expression of VAP-like metalloproteases in the venom gland of three representative pit vipers of the Brazilian territory. By molecular cloning and quantitative real-time polymerase chain reaction, using as calibrator gene the Crotalus durissus terrificus homolog of VAP1, named crotastatin, it is reported here that VAP1/crotastatin-like homologues in the venom gland of Bothrops atrox, C. d. cascavella and Lachesis m. rhombeata are expressed at different levels. Hence, batroxstatins, the crotastatin-like precursors from B. atrox, are expressed 87 times more than crotastatin-1, from C. d. cascavella, and 7.5-fold that lachestatins, from L. m. rhombeata. Moreover, in silico structural analysis of amino acid sequences indicates that batroxstatin-2, crotastatins and lachestatin-1 and -2 which share the archetypal motifs and metal- binding sites of VAP1, are subgrouped in a branch that comprises some apoptosis-inducing toxins.

  16. IFAP Syndrome Is Caused by Deficiency in MBTPS2, an Intramembrane Zinc Metalloprotease Essential for Cholesterol Homeostasis and ER Stress Response

    PubMed Central

    Oeffner, Frank; Fischer, Gayle; Happle, Rudolf; König, Arne; Betz, Regina C.; Bornholdt, Dorothea; Neidel, Ulrike; del Carmen Boente, María; Redler, Silke; Romero-Gomez, Javier; Salhi, Aïcha; Vera-Casaño, Ángel; Weirich, Christian; Grzeschik, Karl-Heinz

    2009-01-01

    Ichthyosis follicularis with atrichia and photophobia (IFAP syndrome) is a rare X-linked, oculocutaneous human disorder. Here, we assign the IFAP locus to the 5.4 Mb region between DXS989 and DXS8019 on Xp22.11-p22.13 and provide evidence that missense mutations exchanging highly conserved amino acids of membrane-bound transcription factor protease, site 2 (MBTPS2) are associated with this phenotype. MBTPS2, a membrane-embedded zinc metalloprotease, activates signaling proteins involved in sterol control of transcription and ER stress response. Wild-type MBTPS2 was able to complement the protease deficiency in Chinese hamster M19 cells as shown by induction of an SRE-regulated reporter gene in transient transfection experiments and by growth of stably transfected cells in media devoid of cholesterol and lipids. These functions were impaired in five mutations as detected in unrelated patients. The degree of diminished activity correlated with clinical severity as noted in male patients. Our findings indicate that the phenotypic expression of IFAP syndrome is quantitatively related to a reduced function of a key cellular regulatory system affecting cholesterol homeostasis and ability to cope with ER stress. PMID:19361614

  17. A phage display selected 7-mer peptide inhibitor of the Tannerella forsythia metalloprotease-like enzyme Karilysin can be truncated to Ser-Trp-Phe-Pro.

    PubMed

    Skottrup, Peter Durand; Sørensen, Grete; Ksiazek, Miroslaw; Potempa, Jan; Riise, Erik

    2012-01-01

    Tannerella forsythia is a gram-negative bacteria, which is strongly associated with the development of periodontal disease. Karilysin is a newly identified metalloprotease-like enzyme, that is secreted from T. forsythia. Karilysin modulates the host immune response and is therefore considered a likely drug target. In this study peptides were selected towards the catalytic domain from Karilysin (Kly18) by phage display. The peptides were linear with low micromolar binding affinities. The two best binders (peptide14 and peptide15), shared the consensus sequence XWFPXXXGGG. A peptide15 fusion with Maltose Binding protein (MBP) was produced with peptide15 fused to the N-terminus of MBP. The peptide15-MBP was expressed in E. coli and the purified fusion-protein was used to verify Kly18 specific binding. Chemically synthesised peptide15 (SWFPLRSGGG) could inhibit the enzymatic activity of both Kly18 and intact Karilysin (Kly48). Furthermore, peptide15 could slow down the autoprocessing of intact Kly48 to Kly18. The WFP motif was important for inhibition and a truncation study further demonstrated that the N-terminal serine was also essential for Kly18 inhibition. The SWFP peptide had a Ki value in the low micromolar range, which was similar to the intact peptide15. In conclusion SWFP is the first reported inhibitor of Karilysin and can be used as a valuable tool in structure-function studies of Karilysin.

  18. Expression of the Matrix Metalloproteases 2, 14, 24, and 25 and Tissue Inhibitor 3 as Potential Molecular Markers in Advanced Human Gastric Cancer

    PubMed Central

    de la Peña, Sol; Sampieri, Clara Luz; Ochoa-Lara, Mariana; León-Córdoba, Kenneth; Remes-Troche, José María

    2014-01-01

    Background. During progression of gastric cancer (GC), degradation of the extracellular matrix is mediated by the matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs): changes in the expression of these have been related to unfavorable prognosis in GC. Objective. To analyze the expression of certain MMPs and TIMPs in chronic superficial gastritis (SG) and GC. Methods. The expression of MMPs and TIMPs was determined using qRT-PCR; the expression was classified, using threshold cycle (CT) values, as very high (CT ≤ 25), high (CT = 26–30), moderate (CT = 31–35), low (CT = 36–39), or not detected (CT = 40). Strength of association was estimated between the proteins, which were detected by Western blot, and the risk of developing GC. Results. We found a high expression of MMP1, MMP2, MMP14, TIMP1, and TIMP3; moderate one of MMP9 and MMP25, and low one of MMP13 and MMP24 in both tissues. In absolute mRNA levels, significant differences were found in expression of MMP2, MMP24, and MMP25, which are overexpressed in GC compared with SG. The presence of the proteins MMP-14 and TIMP-3 was associated with the risk of developing GC. Conclusions. We consider that MMP2, MMP24, and MMP25 and the proteins MMP-14 and TIMP-3 could be candidates for prognostic molecular markers in GC. PMID:24669030

  19. Linking Temporal Changes in Bacterial Community Structures with the Detection and Phylogenetic Analysis of Neutral Metalloprotease Genes in the Sediments of a Hypereutrophic Lake

    PubMed Central

    Tsuboi, Shun; Yamamura, Shigeki; Imai, Akio; Satou, Takayuki; Iwasaki, Kazuhiro

    2014-01-01

    We investigated spatial and temporal variations in bacterial community structures as well as the presence of three functional proteolytic enzyme genes in the sediments of a hypereutrophic freshwater lake in order to acquire an insight into dynamic links between bacterial community structures and proteolytic functions. Bacterial communities determined from 16S rRNA gene clone libraries markedly changed bimonthly, rather than vertically in the sediment cores. The phylum Firmicutes dominated in the 4–6 cm deep sediment layer sample after August in 2007, and this correlated with increases in interstitial ammonium concentrations (p < 0.01). The Firmicutes clones were mostly composed of the genus Bacillus. npr genes encoding neutral metalloprotease, an extracellular protease gene, were detected after the phylum Firmicutes became dominant. The deduced Npr protein sequences from the retrieved npr genes also showed that most of the Npr sequences used in this study were closely related to those of the genus Bacillus, with similarities ranging from 61% to 100%. Synchronous temporal occurrences of the 16S rRNA gene and Npr sequences, both from the genus Bacillus, were positively associated with increases in interstitial ammonium concentrations, which may imply that proteolysis by Npr from the genus Bacillus may contribute to the marked increases observed in ammonium concentrations in the sediments. Our results suggest that sedimentary bacteria may play an important role in the biogeochemical nitrogen cycle of freshwater lakes. PMID:25130992

  20. Analysis of Botulinum Neurotoxin Serotype A Metalloprotease Inhibitors: Analogs of a Chemotype for Therapeutic Development in the Context of a Three-Zone Pharmacophore

    PubMed Central

    Burnett, James C.; Li, Bing; Pai, Ramdas; Cardinale, Steven C.; Butler, Michelle M.; Peet, Norton P.; Moir, Donald; Bavari, Sina; Bowlin, Terry

    2010-01-01

    Botulinum neurotoxins (BoNTs), and in particular serotype A, are the most poisonous of known biological substances, and are responsible for the flaccid paralysis of the disease state botulism. Because of the extreme toxicity of these enzymes, BoNTs are considered highest priority biothreat agents. To counter BoNT serotype A (BoNT/A) poisoning, the discovery and development of small molecule, drug-like inhibitors as post-intoxication therapeutic agents has been/is being pursued. Specifically, we are focusing on inhibitors of the BoNT/A light chain (LC) (ie, a metalloprotease) subunit, since such compounds can enter neurons and provide post-intoxication protection of the enzyme target substrate. To aid/facilitate this drug development effort, a pharmacophore for inhibition of the BoNT/A LC subunit was previously developed, and is continually being refined via the incorporation of novel and diverse inhibitor chemotypes. Here, we describe several analogs of a promising therapeutic chemotype in the context of the pharmacophore for BoNT/A LC inhibition. Specifically, we describe: 1) the pharmacophoric ‘fits’ of the analogs and how these ‘fits’ rationalize the in vitro inhibitory potencies of the analogs and 2) pharmacophore refinement via the inclusion of new components from the most potent of the presented analogs. PMID:21103387

  1. A 24-kDa cloned zinc metalloprotease from Actinobacillus pleuropneumoniae is common to all serotypes and cleaves actin in vitro.

    PubMed Central

    García-Cuéllar, C; Montañez, C; Tenorio, V; Reyes-Esparza, J; Durán, M J; Negrete, E; Guerrero, A; de la Garza, M

    2000-01-01

    Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes proteases that degrade porcine hemoglobin and IgA in vitro. To further characterize A. pleuropneumoniae proteases, we constructed a genomic library expressed in Escherichia coli DH5alpha, and selected a clone that showed proteolytic activity. The recombinant plasmid carries an 800-base pair A. pleuropneumoniae gene sequence that.codes for a 24-kDa polypeptide. A 350-base pair PstI fragment from the sequence hybridized at high stringency with DNA from 12 serotypes of A. pleuropneumoniae, but not with DNA from Actinobacillus suis, Haemophilus parasuis, Pasteurella haemolytica, Pasteurella multocida A or D, or E. coli DH5alpha, thus showing specificity for A. pleuropneumoniae. The expressed polypeptide was recognized as an antigen by convalescent-phase pig sera. Furthermore, a polyclonal antiserum developed against the purified polypeptide recognized an A. pleuropneumoniae oligomeric protein in both crude-extract and cell-free culture media. This recombinant polypeptide cleaved azocoll, gelatin, and actin. Inhibition of the proteolytic activity by diethylpyrocarbonate suggests that this polypeptide is a zinc metalloprotease. Images Figure 1. Figure 2. Figure 3. Figure 4. Figure 6. Figure 7. PMID:10805246

  2. Single particle tracking of ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type-1 repeats) molecules on endothelial von Willebrand factor strings.

    PubMed

    De Ceunynck, Karen; Rocha, Susana; De Meyer, Simon F; Sadler, J Evan; Uji-i, Hiroshi; Deckmyn, Hans; Hofkens, Johan; Vanhoorelbeke, Karen

    2014-03-28

    von Willebrand factor (VWF) strings are removed from the endothelial surface by ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type-1 repeats)-mediated proteolysis. To visualize how single ADAMTS13 molecules bind to these long strings, we built a customized single molecule fluorescence microscope and developed single particle tracking software. Extensive analysis of over 6,000 single inactive ADAMTS13(E225Q) enzymes demonstrated that 20% of these molecules could be detected in at least two consecutive 60-ms frames and followed two types of trajectories. ADAMTS13(E225Q) molecules either decelerated in the vicinity of VWF strings, whereas sometimes making brief contact with the VWF string before disappearing again, or readily bound to the VWF strings and this for 120 ms or longer. These interactions were observed at several sites along the strings. Control experiments using an IgG protein revealed that only the second type of trajectory reflected a specific interaction of ADAMTS13 with the VWF string. In conclusion, we developed a dedicated single molecule fluorescence microscope for detecting single ADAMTS13 molecules (nm scale) on their long, flow-stretched VWF substrates (μm scale) anchored on living cells. Comprehensive analysis of all detected enzymes showed a random interaction mechanism for ADAMTS13 with many available binding sites on the VWF strings.

  3. An extracellular serine protease produced by Vibrio vulnificus NCIMB 2137, a metalloprotease-gene negative strain isolated from a diseased eel.

    PubMed

    Miyoshi, Shin-Ichi; Wang, Jiyou; Katoh, Keizo; Senoh, Mitsutoshi; Mizuno, Tamaki; Maehara, Yoko

    2012-04-01

    Vibrio vulnificus is a ubiquitous estuarine microorganism but causes fatal systemic infections in immunocompromised humans, cultured eels or shrimps. An extracellular metalloprotease VVP/VvpE has been reported to be a potential virulence factor of the bacterium; however, a few strains isolated from a diseased eel or shrimp were recently found to produce a serine protease termed VvsA, but not VVP/VvpE. In the present study, we found that these strains had lost the 80 kb genomic region including the gene encoding VVP/VvpE. We also purified VvsA from the culture supernatant through ammonium sulfate fractionation, gel filtration and ion-exchange column chromatography, and the enzyme was demonstrated to be a chymotrypsin-like protease, as well as those from some vibrios. The gene vvsA was shown to constitute an operon with a downstream gene vvsB, and several Vibrio species were found to have orthologues of vvsAB. These findings indicate that the genes vvp/vvpE and vvsAB might be mobile genetic elements.

  4. Self-cleavage of Human CLCA1 Protein by a Novel Internal Metalloprotease Domain Controls Calcium-activated Chloride Channel Activation*♦

    PubMed Central

    Yurtsever, Zeynep; Sala-Rabanal, Monica; Randolph, David T.; Scheaffer, Suzanne M.; Roswit, William T.; Alevy, Yael G.; Patel, Anand C.; Heier, Richard F.; Romero, Arthur G.; Nichols, Colin G.; Holtzman, Michael J.; Brett, Tom J.

    2012-01-01

    The chloride channel calcium-activated (CLCA) family are secreted proteins that regulate both chloride transport and mucin expression, thus controlling the production of mucus in respiratory and other systems. Accordingly, human CLCA1 is a critical mediator of hypersecretory lung diseases, such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis, that manifest mucus obstruction. Despite relevance to homeostasis and disease, the mechanism of CLCA1 function remains largely undefined. We address this void by showing that CLCA proteins contain a consensus proteolytic cleavage site recognized by a novel zincin metalloprotease domain located within the N terminus of CLCA itself. CLCA1 mutations that inhibit self-cleavage prevent activation of calcium-activated chloride channel (CaCC)-mediated chloride transport. CaCC activation requires cleavage to unmask the N-terminal fragment of CLCA1, which can independently gate CaCCs. Gating of CaCCs mediated by CLCA1 does not appear to involve proteolytic cleavage of the channel because a mutant N-terminal fragment deficient in proteolytic activity is able to induce currents comparable with that of the native fragment. These data provide both a mechanistic basis for CLCA1 self-cleavage and a novel mechanism for regulation of chloride channel activity specific to the mucosal interface. PMID:23112050

  5. Self-cleavage of human CLCA1 protein by a novel internal metalloprotease domain controls calcium-activated chloride channel activation.

    PubMed

    Yurtsever, Zeynep; Sala-Rabanal, Monica; Randolph, David T; Scheaffer, Suzanne M; Roswit, William T; Alevy, Yael G; Patel, Anand C; Heier, Richard F; Romero, Arthur G; Nichols, Colin G; Holtzman, Michael J; Brett, Tom J

    2012-12-07

    The chloride channel calcium-activated (CLCA) family are secreted proteins that regulate both chloride transport and mucin expression, thus controlling the production of mucus in respiratory and other systems. Accordingly, human CLCA1 is a critical mediator of hypersecretory lung diseases, such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis, that manifest mucus obstruction. Despite relevance to homeostasis and disease, the mechanism of CLCA1 function remains largely undefined. We address this void by showing that CLCA proteins contain a consensus proteolytic cleavage site recognized by a novel zincin metalloprotease domain located within the N terminus of CLCA itself. CLCA1 mutations that inhibit self-cleavage prevent activation of calcium-activated chloride channel (CaCC)-mediated chloride transport. CaCC activation requires cleavage to unmask the N-terminal fragment of CLCA1, which can independently gate CaCCs. Gating of CaCCs mediated by CLCA1 does not appear to involve proteolytic cleavage of the channel because a mutant N-terminal fragment deficient in proteolytic activity is able to induce currents comparable with that of the native fragment. These data provide both a mechanistic basis for CLCA1 self-cleavage and a novel mechanism for regulation of chloride channel activity specific to the mucosal interface.

  6. Caveolin-1-dependent activation of the metalloprotease TACE/ADAM17 by TGF-β in hepatocytes requires activation of Src and the NADPH oxidase NOX1.

    PubMed

    Moreno-Càceres, Joaquim; Mainez, Jèssica; Mayoral, Rafael; Martín-Sanz, Paloma; Egea, Gustavo; Fabregat, Isabel

    2016-04-01

    Transforming growth factor-β (TGF-β) plays a dual role in hepatocytes, inducing both pro- and anti-apoptotic responses, the balance between which decides cell fate. Survival signals are mediated by the epidermal growth factor receptor (EGFR) pathway, which is activated by TGF-β. We have previously shown that caveolin-1 (CAV1) is required for activation of the metalloprotease tumour necrosis factor (TNF)-α-converting enzyme/a disintegrin and metalloproteinase 17 (TACE/ADAM17), and hence transactivation of the EGFR pathway. The specific mechanism by which TACE/ADAM17 is activated has not yet been determined. Here we show that TGF-β induces phosphorylation of sarcoma kinase (Src) in hepatocytes, a process that is impaired in Cav1(-/-) hepatocytes, coincident with a decrease in phosphorylated Src in detergent-resistant membrane fractions. TGF-β-induced activation of TACE/ADAM17 and EGFR phosphorylation were blocked using the Src inhibitor PP2. Cav1(+/+) hepatocytes showed early production of reactive oxygen species (ROS) induced by TGF-β, which was not seen in Cav1(-/-) cells. Production of ROS was inhibited by both the NADPH oxidase 1 (NOX1) inhibitor STK301831 and NOX1 knock-down, which also impaired TACE/ADAM17 activation and thus EGFR phosphorylation. Finally, neither STK301831 nor NOX1 silencing impaired Src phosphorylation, but PP2 blocked early ROS production, showing that Src is involved in NOX1 activation. As expected, inhibition of Src or NOX1 increased TGF-β-induced cell death in Cav1(+/+) cells. In conclusion, CAV1 is required for TGF-β-mediated activation of TACE/ADAM17 through a mechanism that involves phosphorylation of Src and NOX1-mediated ROS production.

  7. The hookworm tissue inhibitor of metalloproteases (Ac-TMP-1) modifies dendritic cell function and induces generation of CD4 and CD8 suppressor T cells.

    PubMed

    Cuéllar, Carmen; Wu, Wenhui; Mendez, Susana

    2009-05-19

    Hookworm infection is a major cause of disease burden for humans. Recent studies have described hookworm-related immunosuppression in endemic populations and animal models. A Tissue Inhibitor of Metalloproteases (Ac-TMP-1) has been identified as one of the most abundant proteins released by the adult parasite. We investigated the effect of recombinant Ac-TMP-1 on dendritic cell (DC) and T cell function. Splenic T cells from C57BL/6 mice injected with Ac-TMP-1 showed reduced proliferation to restimulation with anti CD3 or bystander antigens such as OVA. Incubation of bone marrow-derived DCs with Ac-TMP-1 decreased MHC Class I and, especially, Class II expression but increased CD86 and IL-10 expression. Co-incubation of splenic T cells with DCs pulsed with Ac-TMP-1 induced their differentiation into CD4+ and, particularly, CD8+ CD25+Foxp3+ T cells that expressed IL-10. These cells were able to suppress proliferation of naïve and activated CD4+ T cells by TGF-Beta-dependent (CD4+ suppressors) or independent (CD8+ suppressors) mechanisms. Priming of DCs with non-hookworm antigens, such as OVA, did not result in the generation of suppressor T cells. These data indicate that Ac-TMP-1 initiates the development of a regulatory response through modifications in DC function and generation of suppressor T cells. This is the first report to propose a role of suppressor CD8+ T cells in gastrointestinal helminthic infections.

  8. Characterization of a metalloprotease from ovine chromaffin granules which cleaves a proenkephalin fragment (BAM12P) at a single arginine residue.

    PubMed Central

    Tezapsidis, N; Parish, D C

    1994-01-01

    A metalloprotease has been identified in ovine chromaffin granules which cleaves the proenkephalin fragment BAM12P to produce adrenorphin-Gly. This cleavage occurs at a single arginine residue and is an intermediate step in the production of the opiate adrenorphin in vivo. The identity of the product was confirmed by reverse-phase and ion-exchange chromatography. The adrenorphin-Gly-generating enzyme (AGE) was determined by chromatofocusing to have a pI value of 5.2 and bound strongly to a metal-chelate affinity column. After purification by gel-filtration and ion-exchange chromatography AGE was free of contaminating activities, as cleavage of radiolabelled BAM12P generated a single product as judged by reverse-phase and ion-exchange chromatography. The enzyme has a molecular mass of approx. 45 kDa and a pH optimum of 8.6 in Mops, Taps and Hepes buffers, but was inhibited by phosphate buffers. It was inhibited by micromolar concentrations of copper and zinc ions, but not by millimolar concentrations of calcium or manganese ions. The addition of BAM22P, dynorphin 1-13 or dynorphin 1-8 to the incubation mixture inhibited the cleavage of radiolabelled BAM12P. The cleavage was also inhibited by the presence of catecholamines at concentrations similar to those found within the chromaffin granule. This may explain the known effect of reserpine on chromaffin cells of reducing catecholamine levels and simultaneously increasing adrenorphin levels. It may also indicate a function for AGE and adrenorphin as reporters of intragranular conditions. Images Figure 1 PMID:8043007

  9. Adamts5, the gene encoding a proteoglycan-degrading metalloprotease, is expressed by specific cell lineages during mouse embryonic development and in adult tissues.

    PubMed

    McCulloch, Daniel R; Le Goff, Carine; Bhatt, Sumantha; Dixon, Laura J; Sandy, John D; Apte, Suneel S

    2009-06-01

    The secreted metalloprotease ADAMTS5 is implicated in destruction of the cartilage proteoglycan aggrecan in arthritis, but its physiological functions are unknown. Its expression profile during embryogenesis and in adult tissues is therefore of considerable interest. beta-Galactosidase (beta-gal) histochemistry, enabled by a LacZ cassette inserted in the Adamts5 locus, and validated by in situ hybridization with an Adamts5 cRNA probe and ADAMTS5 immunohistochemistry, was used to profile Adamts5 expression during mouse embryogenesis and in adult mouse tissues. Embryonic expression was scarce prior to 11.5 days of gestation (E11.5) and noted only in the floor plate of the developing brain at E 9.5. After E11.5 there was continued expression in brain, especially in the choroid plexus, peripheral nerves, dorsal root ganglia, cranial nerve ganglia, spinal and cranial nerves, and neural plexuses of the gut. In addition to nerves, developing limbs have Adamts5 expression in skeletal muscle (from E13.5), tendons (from E16.5), and inter-digital mesenchyme of the developing autopod (E13.5-15.5). In adult tissues, there is constitutive Adamts5 expression in arterial smooth muscle cells, mesothelium lining the peritoneal, pericardial and pleural cavities, smooth muscle cells in bronchi and pancreatic ducts, glomerular mesangial cells in the kidney, dorsal root ganglia, and in Schwann cells of the peripheral and autonomic nervous system. Expression of Adamts5 during neuromuscular development and in smooth muscle cells coincides with the broadly distributed proteoglycan versican, an ADAMTS5 substrate. These observations suggest the major contexts in which developmental and physiological roles could be sought for this protease.

  10. Evidence for Cleavage of the Metalloprotease Vsm from Vibrio splendidus Strain JZ6 by an M20 Peptidase (PepT-like Protein) at Low Temperature

    PubMed Central

    Liu, Rui; Qiu, Limei; Cheng, Qi; Zhang, Huan; Wang, Lingling; Song, Linsheng

    2016-01-01

    Metalloprotease Vsm is a major extracellular virulence factor of Vibrio splendidus. The toxicity of Vsm from V. splendidus strain JZ6 has been characterized, and production of this virulence factor proved to be temperature-regulated. The present study provides evidence that two forms (JZE1 and JZE2) of Vsm protein exist in extracellular products (ECPs) of strain JZ6, and a significant conversion of these two forms was detected by SDS-PAGE and immunoblotting analyses of samples obtained from cells grown at 4, 10, 16, 20, 24, and 28°C. Mass spectroscopy confirmed that JZE1 was composed only of the peptidase_M4 domain of Vsm, and JZE2 contained both the PepSY domain and the peptidase_M4 domain. An M20 peptidase T-like protein (PepTL) was screened from the transcriptome data of strain JZ6, which was considered as a crucial molecule to produce the active Vsm (JZE1) by cleavage of the propeptide. Similar to that of Vsm, PepTL mRNA accumulation was highest at 4°C (836.82-fold of that at 28°C), decreased with increasing of temperature and reached its lowest level at 28°C. Deletion of the gene encoding the PepTL resulted in a mutant strain that did not produce the JZE1 cleavage product. The peptidase activity of PepTL recombinant protein (rPepTL) was confirmed by cleaving the Vsm in ECPs with an in vitro degradation reaction. These results demonstrate that PepTL participates in activating Vsm in strain JZ6 by proteolytic cleavage at low temperature. PMID:27826294

  11. A Disintegrin and Metalloprotease (ADAM) 10 and ADAM17 Are Major Sheddases of T Cell Immunoglobulin and Mucin Domain 3 (Tim-3)*

    PubMed Central

    Möller-Hackbarth, Katja; Dewitz, Christin; Schweigert, Olga; Trad, Ahmad; Garbers, Christoph; Rose-John, Stefan; Scheller, Jürgen

    2013-01-01

    T cell immunoglobulin and mucin domain 3 (Tim-3) dampens the response of CD4+ and CD8+ effector T cells via induction of cell death and/or T cell exhaustion and enhances the ability of macrophages to clear pathogens via binding to galectin 9. Here we provide evidence that human Tim-3 is a target of A disintegrin and metalloprotease (ADAM)-mediated ectodomain shedding resulting in a soluble form of Tim-3. We identified ADAM10 and ADAM17 as major sheddases of Tim-3 as shown by ADAM-specific inhibitors and the ADAM10 pro-domain in HEK293 cells and ADAM10/ADAM17-deficient murine embryonic fibroblasts. PMA-induced shedding of Tim-3 was abrogated by deletion of amino acids Glu181–Asp190 of the stalk region and Tim-3 lacking the intracellular domain was not efficiently cleaved after PMA stimulation. Surprisingly, a single lysine residue within the intracellular domain rescues shedding of Tim-3. Shedding of endogenous Tim-3 was found in primary human CD14+ monocytes after PMA and ionomycin stimulation. Importantly, the recently described down-regulation of Tim-3 from Toll-like receptor-activated CD14+ monocytes was caused by ADAM10- and ADAM17-mediated shedding. Inhibition of Tim-3 shedding from lipopolysaccharide-induced monocytes did not influence lipopolysaccharide-induced TNFα and IL-6 but increases IL-12 expression. In summary, we describe Tim-3 as novel target for ADAM-mediated ectodomain shedding and suggest a role of Tim-3 shedding in TLR-mediated immune responses of CD14+ monocytes. PMID:24121505

  12. Isolation, characterization and molecular three-dimensional structural predictions of metalloprotease from a phytopathogenic fungus, Alternaria solani (Ell. and Mart.) Sor.

    PubMed

    Chandrasekaran, Murugesan; Chandrasekar, Raman; Chun, Se-Chul; Sathiyabama, Muthukrishnan

    2016-08-01

    The present study aims at isolation, identification, characterization and prediction of three-dimensional molecular architecture of a proteolytic enzyme from the early blight pathogen, Alternaria solani which are hypothesized to be a marker of phytopathogenicity. Maximum enzyme production by A. solani was observed in Czapex's Dox broth amended with 2% (w/v) casein than other inducer amendments. Results indicate that the enzyme remained highly active in a pH range of 7.0-10.0 and a temperature range of 45-50°C. The enzyme was strongly inhibited by EDTA, whereas phenylmethylsulfonyl fluoride and monovalent cations (Na(+), K(+)) had little effect. Metal ions such as MgSO4, CaCl2, KCl at 10 mM concentration showed a stimulatory effect (>85%) on protease activity. Matrix-assisted laser desorption and ionization time of flight/mass spectrometry analysis of partially purified enzyme revealed the presence of protease belonging to a keratinolytic protein (metalloprotease) of exopeptidase nature. Putative A. solani keratinolytic enzyme (AsK) is made up of 216 amino acid residues with molecular weight (MW) 24.5 kDa, having a molecular formula of C1094H1704N290O342S4. Ramachandran plot analysis of the protein residues falling into the most favored secondary structures was observed at 84.2%. The major protein structural blocks, 2-β-sheets, and 9-α-helices have a greater tendency to be conserved during the evolutionary process than do mere sequences of amino acids. Besides, AsK, model prediction showed the presence of a Zinc atom at helix regions (Helix 3, 6, 7: His(57), His(130), His(169), and Cys(123)). Thus, it can be concluded that the major proteinases of AsK are divalent cation-requiring metalloproteinases and make them potential targets of protease inhibitors designing.

  13. Effects of chondroitin and glucosamine sulfate in a dietary bar formulation on inflammation, interleukin-1beta, matrix metalloprotease-9, and cartilage damage in arthritis.

    PubMed

    Chou, May M; Vergnolle, Nathalie; McDougall, Jason J; Wallace, John L; Marty, Stephanie; Teskey, Val; Buret, Andre G

    2005-04-01

    This study examined the effects of chondroitin sulfate (CS) alone and CS plus glucosamine sulfate (GS) in a dietary bar formulation on inflammatory parameters of adjuvant-induced arthritis and on the synthesis of interleukin-1beta (IL-1beta) and matrix metalloprotease-9 (MMP-9). Following 25 days pretreatment with dietary bars containing either CS alone, CS plus GS, or neither CS nor GS, rats were either sham injected or injected with Freund's complete adjuvant into the tail vein. Rats were fed their respective bars for another 17 days after inoculation. Parameters of disease examined included clinical score (combination of joint temperature, edema, hyperalgesia, and standing and walking limb function), incidence of disease, levels of IL-1beta in the serum and paw joints, levels of MMP-9 in the paw joints, paw joint histology, and joint cartilage thickness. Treatment with CS plus GS, but not CS alone, significantly reduced clinical scores, incidences of disease, joint temperatures, and joint and serum IL-1beta levels. Treatment with CS alone and CS plus GS inhibited the production of edema and prevented raised levels of joint MMP-9 associated with arthritis. Similarly, CS alone and CS plus GS treatment also prevented the development of cartilage damage associated with arthritis. Combination CS plus GS treatment in a dietary bar formulation ameliorates clinical, inflammatory, and histologic parameters of adjuvant-induced arthritis. The benefits of CS and GS in combination are more pronounced than those of CS alone. The reduction of arthritic disease by CS plus GS is associated with a reduction of IL-1beta and MMP-9 synthesis.

  14. Acute morphine activates satellite glial cells and up-regulates IL-1β in dorsal root ganglia in mice via matrix metalloprotease-9

    PubMed Central

    2012-01-01

    Background Activation of spinal cord glial cells such as microglia and astrocytes has been shown to regulate chronic opioid-induced antinociceptive tolerance and hyperalgesia, due to spinal up-regulation of the proinflammatory cytokines such as interleukin-1 beta (IL-1β). Matrix metalloprotease-9 (MMP-9) has been implicated in IL-1β activation in neuropathic pain. However, it is unclear whether acute opioid treatment can activate glial cells in the peripheral nervous system. We examined acute morphine-induced activation of satellite glial cells (SGCs) and up-regulation of IL-1β in dorsal root ganglia (DRGs), and further investigated the involvement of MMP-9 in these opioid-induced peripheral changes. Results Subcutaneous morphine injection (10 mg/kg) induced robust peripheral glial responses, as evidenced by increased GFAP expression in DRGs but not in spinal cords. The acute morphine-induced GFAP expression is transient, peaking at 2 h and declining after 3 h. Acute morphine treatment also increased IL-1β immunoreactivity in SGCs and IL-1β activation in DRGs. MMP-9 and GFAP are expressed in DRG neurons and SGCs, respectively. Confocal analysis revealed a close proximity of MMP-9 and GFAP immunostaining. Importantly, morphine-induced DRG up-regulation of GFAP expression and IL-1β activation was abolished after Mmp9 deletion or naloxone pre-treatment. Finally, intrathecal injections of IL-1β-selective siRNA not only reduced DRG IL-1β expression but also prolonged acute morphine-induced analgesia. Conclusions Acute morphine induces opioid receptors- and MMP-9-dependent up-regulation of GFAP expression and IL-1β activation in SGCs of DRGs. MMP-9 could mask and shorten morphine analgesia via peripheral neuron-glial interactions. Targeting peripheral glial activation might prolong acute opioid analgesia. PMID:22439811

  15. Cloning and characterization of a cDNA coding for Astacus embryonic astacin, a member of the astacin family of metalloproteases from the crayfish Astacus astacus.

    PubMed

    Geier, G; Zwilling, R

    1998-05-01

    The astacin family of zinc endopeptidases was named after the digestive enzyme astacin isolated from the crayfish Astacus astacus. Employing a reverse transcription/PCR strategy with degenerate oligonucleotide primers specific for two signature seqences of the astacin family, we have isolated a 1602-bp cDNA from embryos of developing A. astacus eggs, which was designated Astacus embryonic astacin (AEA). This cDNA was found to code for an astacin-like protease domain which accounts for the N-terminal half of the predicted protein. The C-terminal half mainly consists of two complement subcomponent C1r/C1s/embryonic sea urchin protein Uegf/bone morphogenetic protein 1 (CUB) domains. The metalloprotease domain displays an amino acid sequence identity of 42% with astacin. A higher sequence similarity was found to astacin family members that act as hatching enzymes in different species, e.g. chorioallantoic membrane protein 1 (CAM-1; from quail) and Xenopus hatching enzyme (formerly UVS.2), both of which show 54% identity, and high and low choriolytic enzymes (HCE and LCE) from the teleost Oryzias latipes (52% and 48% identity, respectively). A relationship to astacin-like hatching enzymes is further supported by a phylogenetic analysis of the protease domains. Expression of AEA mRNA in developing embryos was found to be restricted to unhatched juveniles (larvae) during the last 8 days before hatching. AEA transcripts could not be detected in various tissues of adult animals or in eggs and embryos from an earlier developmental stage. AEA expression starts about 8 days prior to hatching, followed by a strong (18-fold) induction with a maximum at day 4 before hatching. Newly hatched juveniles were found not to express the AEA mRNA.

  16. TH1/TH2 cytokine profile, metalloprotease-9 activity and hormonal status in pregnant rheumatoid arthritis and systemic lupus erythematosus patients

    PubMed Central

    MUÑOZ-VALLE, J F; VÁZQUEZ-DEL MERCADO, M; GARCÍA-IGLESIAS, T; OROZCO-BAROCIO, G; BERNARD-MEDINA, G; MARTÍNEZ-BONILLA, G; BASTIDAS-RAMÍREZ, B E; NAVARRO, A D; BUENO, M; MARTÍNEZ-LÓPEZ, E; BEST-AGUILERA, C R; KAMACHI, M; ARMENDÁRIZ-BORUNDA, J

    2003-01-01

    During the course of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), several immune and neuroendocrine changes associated with pregnancy may exert positive (amelioration) or negative (exacerbation) effects on the clinical outcome. In order to shed light on the mechanisms underlying these responses, we performed a prospective longitudinal study in RA and SLE pregnant women, including healthy pregnant women as a control group. Cytokine messenger RNA (mRNA) expression assessed by quantitative competitive polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), cytokine levels and lymphocyte proliferation responses (LPR) following phytohaemagglutinin (PHA) stimulation of PBMC, plasma metalloprotease-9 activity (MMP-9) and hormonal status during pregnancy were determined. TNFa was the most abundant cytokine mRNA expressed in PBMC in all groups studied (healthy pregnant women, RA and SLE pregnant patients). However, a general TH2 response reflected by high IL-10 levels was found in RA, as well as SLE, patients. A significant change in IFN-γ was observed in RA patients but only during the first trimester of pregnancy. This compared with a major TH1 response in healthy pregnant women. Interestingly, our study showed a homogeneous hormonal pattern in RA and SLE patients. Although decreased cortisol levels were observed in all patients studied, this is possibly related to the remission of disease activity status brought about by steroid treatment before and during pregnancy. In summary, we suggest that complex immune and hormonal networks are involved in pregnancy and that rheumatic diseases are very dynamic immune processes that cannot be described with a clear-cut cytokine profile. Furthermore, the observations in this study may reflect treatment-related immune effects more than those associated with disease. PMID:12562402

  17. Transcription factor KLF6 upregulates expression of metalloprotease MMP14 and subsequent release of soluble endoglin during vascular injury.

    PubMed

    Gallardo-Vara, Eunate; Blanco, Francisco J; Roqué, Mercè; Friedman, Scott L; Suzuki, Toru; Botella, Luisa M; Bernabeu, Carmelo

    2016-04-01

    After endothelial injury, the transcription factor Krüppel-like factor 6 (KLF6) translocates into the cell nucleus to regulate a variety of target genes involved in angiogenesis, vascular repair and remodeling, including components of the membrane transforming growth factor beta (TGF-β) receptor complex such as endoglin and activin receptor-like kinase 1. The membrane metalloproteinase 14 (MMP14 or MT1-MMP) targets endoglin to release soluble endoglin and is involved in vascular inflammation and endothelial tubulogenesis. However, little is known about the regulation of MMP14 expression during vascular wounding. In vitro denudation of monolayers of human endothelial cell monolayers leads to an increase in the KLF6 gene transcriptional rate, followed by an upregulation of MMP14 and release of soluble endoglin. Concomitant with this process, MMP14 co-localizes with endoglin in the sprouting endothelial cells surrounding the wound border. MMP14 expression at mRNA and protein levels is increased by ectopic KLF6 and downregulated by KLF6 suppression in cultured endothelial cells. Moreover, after wire-induced endothelial denudation, Klf6 (+/-) mice show lower levels of MMP14 in their vasculature compared with their wild-type siblings. Ectopic cellular expression of KLF6 results in an increased transcription rate of MMP14, and chromatin immunoprecipitation assays show that KLF6 interacts with MMP14 promoter in ECs, this interaction being enhanced during wound healing. Furthermore, KLF6 markedly increases the transcriptional activity of different reporter constructs of MMP14 gene promoter. These results suggest that KLF6 regulates MMP14 transcription and is a critical player of the gene expression network triggered during endothelial repair.

  18. A phenotype from tumor stroma based on the expression of metalloproteases and their inhibitors, associated with prognosis in breast cancer

    PubMed Central

    Eiró, Noemí; Fernandez-Garcia, Belen; Vázquez, Julio; del Casar, José M; González, Luis O; Vizoso, Francisco J

    2015-01-01

    The objective of the present work was to evaluate the impact of the phenotype of both mononuclear inflammatory cells (MICs) and cancer-associated fibroblast (CAFs) in early breast cancer patients, specifically assessed as to their expression of MMP/TIMP relative to their position within the tumor (i.e., localization at the tumor center or invasive front) and the occurrence of distant metastases.. An immunohistochemical study was performed using tissue arrays and specific antibodies against matrix metalloproteinase (MMP)−1, −2, −7, −9, −11, −13 and −14, tissue inhibitors of metalloproteinase (TIMP)−1, −2 and −3, both at tumor center and at invasive front, in 107 patients with primary ductal invasive breast tumors. Data were analyzed by unsupervised hierarchical clustering analysis. Our results indicated that MMP-11 expression by MICs, and TIMP-2 expression by CAFs at either the tumor center or the invasive front, were the most potent independent prognostic factors for predicting the clinical outcome of patients. Using the unsupervised hierarchical clustering analysis, we found well-defined clusters of cases identifying subgroups of tumors showing a high molecular profile of MMPs/TIMPs expression by stromal cells (CAFs and MICs), both at the tumor center and at the invasive front, which were strongly associated with a higher prevalence of distant metastasis. In addition, we found combinations of these clusters defining subpopulations of breast carcinomas differing widely in their clinical outcome. The results presented here identify biologic markers useful to categorize patients into different subgroups based on their tumor stroma, which may contribute to improved understanding of the prognosis of breast cancer patients. PMID:26140253

  19. Semaphorin7A Promotion of Tumoral Growth and Metastasis in Human Oral Cancer by Regulation of G1 Cell Cycle and Matrix Metalloproteases: Possible Contribution to Tumoral Angiogenesis

    PubMed Central

    Saito, Tomoaki; Kasamatsu, Atsushi; Ogawara, Katsunori; Miyamoto, Isao; Saito, Kengo; Iyoda, Manabu; Suzuki, Takane; Endo-Sakamoto, Yosuke; Shiiba, Masashi; Tanzawa, Hideki; Uzawa, Katsuhiro

    2015-01-01

    Background Semaphorins (SEMAs) consist of a large family of secreted and membrane-anchored proteins that are important in neuronal pathfinding and axon guidance in selected areas of the developing nervous system. Of them, SEMA7A has been reported to have a chemotactic activity in neurogenesis and to be an immunomodulator; however, little is known about the relevance of SEMA7A in the behaviors of oral squamous cell carcinoma (OSCC). Methods We evaluated SEMA7A expression in OSCC-derived cell lines and primary OSCC samples using quantitative reverse transcriptase-polymerase chain reaction, immunoblotting, and semiquantitative immunohistochemistry (sq-IHC). In addition, SEMA7A knockdown cells (shSEMA7A cells) were used for functional experiments, including cellular proliferation, invasiveness, and migration assays. We also analyzed the clinical correlation between SEMA7A status and clinical behaviors in patients with OSCC. Results SEMA7A mRNA and protein were up-regulated significantly (P<0.05) in OSCC-derived cell lines compared with human normal oral keratinocytes. The shSEMA7A cells showed decreased cellular growth by cell-cycle arrest at the G1 phase, resulting from up-regulation of cyclin-dependent kinase inhibitors (p21Cip1 and p27Kip1) and down-regulation of cyclins (cyclin D1, cyclin E) and cyclin-dependent kinases (CDK2, CDK4, and CDK6); and decreased invasiveness and migration activities by reduced secretion of matrix metalloproteases (MMPs) (MMP-2, proMMP-2, pro-MMP-9), and expression of membrane type 1- MMP (MT1-MMP). We also found inactivation of the extracellular regulated kinase 1/2 and AKT pathways, an upstream molecule of cell-cycle arrest at the G1 phase, and reduced secretion of MMPs in shSEMA7A cells. sq-IHC showed that SEMA7A expression in the primary OSCCs was significantly (P = 0.001) greater than that in normal counterparts and was correlated with primary tumoral size (P = 0.0254) and regional lymph node metastasis (P = 0.0002). Conclusion Our

  20. New approach for separating Bacillus subtilis metalloprotease and alpha-amylase by affinity chromatography and for purifying neutral protease by hydrophobic chromatography.

    PubMed

    Lauer, I; Bonnewitz, B; Meunier, A; Beverini, M

    2000-01-14

    Proteases are commonly used in the biscuit and cracker industry as processing aids. They cause moderate hydrolysis of gluten proteins and improve dough rheology to better control product texture and crunchiness. Commercial bacterial proteases are derived from Bacillus fermentation broth. As filtration and ultrafiltration are carried out as the only recovery steps, these preparations contain also alpha-amylase and beta-glucanase as the main side activities. The aim of this study is to purify and characterize the Bacillus subtilis metalloprotease from a commercial preparation, in order to study separately the impact of the protease activity with regards to its functionality on biscuit properties. Purification was achieved by means of affinity chromatography on Cibacron Blue and HIC as a polishing step. Affinity appeared to be the most appropriate matrix for large scale purification while ion exchange chromatography was inefficient in terms of recovery yields. The crude product was first loaded on a Hi Trap Blue column (34 microm, Pharmacia Biotech); elution was carried out with a gradient of NaCl in the presence of 1 mM ZnCl2. This step was only efficient in the presence of Zn cations, because this salt promoted both protease stabilization resulting in high recovery yields and also complexation of amylase units into dimers resulting in amylase retention on the column and a better separation of the 3 activities. Beta-glucanase was mostly non retained on the column and a part was coeluted with the protease. This protease fraction was then loaded on a Resource Phe column (15 microm, Pharmacia Biotech) in a last step of polishing. Elution was carried out with a linear gradient of 100-0% ammonium sulfate 1.3 M; protease was eluted at the beginning of the gradient and well separated from amylase and glucanase trace impurities. The homogeneity of the purified protease was confirmed by SDS-PAGE, which showed that its MW was about 38. pH and temperature optima were also

  1. The binding effectiveness of anti-r-disintegrin polyclonal antibodies against disintegrins and PII and PIII metalloproteases: An immunological survey of type A, B and A + B venoms from Mohave rattlesnakes

    PubMed Central

    Cantú, Esteban; Mallela, Sahiti; Nyguen, Matthew; Báez, Raúl; Parra, Victoria; Johnson, Rachel; Wilson, Kyle; Suntravat, Montamas; Lucena, Sara; Rodríguez-Acosta, Alexis; Sánchez, Elda E.

    2016-01-01

    Snake venoms are known to have different venom compositions and toxicity, but differences can also be found within populations of the same species contributing to the complexity of treatment of envenomated victims. One of the first well-documented intraspecies venom variations comes from the Mohave rattlesnake (Crotalus scutulatus scutulatus). Initially, three types of venoms were described; type A venom is the most toxic as a result of ~45% Mojave toxin in the venom composition, type B lacks the Mojave toxin but contains over 50% of snake venom metalloproteases (SVMPs). Also, type A + B venom contains a combination of Mojave toxin and SVMP. The use of an anti-disintegrin antibody in a simple Enzyme-Linked Immunosorbent Assay (ELISA) can be used to identify the difference between the venoms of the type A, B, and A+B Mohave rattlesnakes. This study implements the use of an anti-recombinant disintegrin polyclonal antibody (ARDPA) for the detection of disintegrins and ADAMs (a disintegrin and metalloproteases) in individual crude snake venoms of Mohave rattlesnakes (Crotalus scutulatus scutulatus) of varying geographical locations. After correlation with Western blots, coagulation activity and LD50 data, it was determined that the antibody allows for a quick and cost-efficient identification of venom types. PMID:27989783

  2. Spatial localization of bacteria controls coagulation of human blood by 'quorum acting'.

    PubMed

    Kastrup, Christian J; Boedicker, James Q; Pomerantsev, Andrei P; Moayeri, Mahtab; Bian, Yao; Pompano, Rebecca R; Kline, Timothy R; Sylvestre, Patricia; Shen, Feng; Leppla, Stephen H; Tang, Wei-Jen; Ismagilov, Rustem F

    2008-12-01

    Blood coagulation often accompanies bacterial infections and sepsis and is generally accepted as a consequence of immune responses. Though many bacterial species can directly activate individual coagulation factors, they have not been shown to directly initiate the coagulation cascade that precedes clot formation. Here we demonstrated, using microfluidics and surface patterning, that the spatial localization of bacteria substantially affects coagulation of human and mouse blood and plasma. Bacillus cereus and Bacillus anthracis, the anthrax-causing pathogen, directly initiated coagulation of blood in minutes when bacterial cells were clustered. Coagulation of human blood by B. anthracis required secreted zinc metalloprotease InhA1, which activated prothrombin and factor X directly (not via factor XII or tissue factor pathways). We refer to this mechanism as 'quorum acting' to distinguish it from quorum sensing--it does not require a change in gene expression, it can be rapid and it can be independent of bacterium-to-bacterium communication.

  3. Local gravitomagnetism

    NASA Astrophysics Data System (ADS)

    Shahid-Saless, Bahman

    1990-10-01

    In a simple two-body system, the gravitomagnetic components of the metric in the local quasi-inertial frame of one of the bodies is calculated. The local geometry in this frame which is freely falling along the geodesic but is directionally fixed with respect to distant stars is primarily defined by the gravitomagnetic components of the local metric. This metric serves to track down the various contributions from the local and distant source and thus provides further insight to the nature of gravitomagnetism. As a result it is shown that in the quasi-inertial frame geodetic precession is a gravitomagnetic phenomenon. Furthermore a connection between local gravitomagnetic effects and Einstein's principle of equivalence is established.

  4. Local Group

    NASA Astrophysics Data System (ADS)

    Mateo, M.; Murdin, P.

    2000-11-01

    Not long after EDWIN HUBBLE established that galaxies are `island universes' similar to our home galaxy, the MILKY WAY, he realized that a few of these external galaxies are considerably closer to us than any others. In 1936 he first coined the term `Local Group' in his famous book The Realm of the Nebulae to identify our nearest galactic neighbors. More than 60 yr later, the galaxies of the Loca...

  5. Cold adaptation of zinc metalloproteases in the thermolysin family from deep sea and arctic sea ice bacteria revealed by catalytic and structural properties and molecular dynamics: new insights into relationship between conformational flexibility and hydrogen bonding.

    PubMed

    Xie, Bin-Bin; Bian, Fei; Chen, Xiu-Lan; He, Hai-Lun; Guo, Jun; Gao, Xiang; Zeng, Yin-Xin; Chen, Bo; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2009-04-03

    Increased conformational flexibility is the prevailing explanation for the high catalytic efficiency of cold-adapted enzymes at low temperatures. However, less is known about the structural determinants of flexibility. We reported two novel cold-adapted zinc metalloproteases in the thermolysin family, vibriolysin MCP-02 from a deep sea bacterium and vibriolysin E495 from an Arctic sea ice bacterium, and compared them with their mesophilic homolog, pseudolysin from a terrestrial bacterium. Their catalytic efficiencies, k(cat)/K(m) (10-40 degrees C), followed the order pseudolysin < MCP-02 < E495 with a ratio of approximately 1:2:4. MCP-02 and E495 have the same optimal temperature (T(opt), 57 degrees C, 5 degrees C lower than pseudolysin) and apparent melting temperature (T(m) = 64 degrees C, approximately 10 degrees C lower than pseudolysin). Structural analysis showed that the slightly lower stabilities resulted from a decrease in the number of salt bridges. Fluorescence quenching experiments and molecular dynamics simulations showed that the flexibilities of the proteins were pseudolysin < MCP-02 < E495, suggesting that optimization of flexibility is a strategy for cold adaptation. Molecular dynamics results showed that the ordinal increase in flexibility from pseudolysin to MCP-02 and E495, especially the increase from MCP-02 to E495, mainly resulted from the decrease of hydrogen-bond stability in the dynamic structure, which was due to the increase in asparagine, serine, and threonine residues. Finally, a model for the cold adaptation of MCP-02 and E495 was proposed. This is the first report of the optimization of hydrogen-bonding dynamics as a strategy for cold adaptation and provides new insights into the structural basis underlying conformational flexibility.

  6. Quantum Locality?

    NASA Astrophysics Data System (ADS)

    Stapp, Henry P.

    2012-05-01

    , in response to Griffiths' challenge, why a putative proof of locality that he has described is flawed.

  7. Local Foods, Local Places Summary Reports

    EPA Pesticide Factsheets

    These summary reports describe Local Foods, Local Places projects in communities across the country, including farmers markets, cooperatives, community gardens, and other food-related enterprises that can boost local economies and drive revitalization.

  8. The alpha and beta subunits of the metalloprotease meprin are expressed in separate layers of human epidermis, revealing different functions in keratinocyte proliferation and differentiation.

    PubMed

    Becker-Pauly, Christoph; Höwel, Markus; Walker, Tatjana; Vlad, Annica; Aufenvenne, Karin; Oji, Vinzenz; Lottaz, Daniel; Sterchi, Erwin E; Debela, Mekdes; Magdolen, Viktor; Traupe, Heiko; Stöcker, Walter

    2007-05-01

    The zinc endopeptidase meprin (EC 3.4.24.18) is expressed in brush border membranes of intestine and kidney tubules, intestinal leukocytes, and certain cancer cells, suggesting a role in epithelial differentiation and cell migration. Here we show by RT-PCR and immunoblotting that meprin is also expressed in human skin. As visualized by immunohistochemistry, the two meprin subunits are localized in separate cell layers of the human epidermis. Meprin alpha is expressed in the stratum basale, whereas meprin beta is found in cells of the stratum granulosum just beneath the stratum corneum. In hyperproliferative epidermis such as in psoriasis vulgaris, meprin alpha showed a marked shift of expression from the basal to the uppermost layers of the epidermis. The expression patterns suggest distinct functions for the two subunits in skin. This assumption is supported by diverse effects of recombinant meprin alpha and beta on human adult low-calcium high-temperature keratinocytes. Here, beta induced a dramatic change in cell morphology and reduced the cell number, indicating a function in terminal differentiation, whereas meprin alpha did not affect cell viability, and may play a role in basal keratinocyte proliferation.

  9. Quantum Locality?

    SciTech Connect

    Stapp, Henry

    2011-11-10

    vagaries that he cites do not upset the proof in question. It is show here in detail why the precise statement of this theorem justifies the specified application of CQT. It is also shown, in response to his challenge, why a putative proof of locality that he has proposed is not valid.

  10. cDNA sequence and chromosomal localization of human enterokinase, the proteolytic activator of trypsinogen.

    PubMed

    Kitamoto, Y; Veile, R A; Donis-Keller, H; Sadler, J E

    1995-04-11

    Enterokinase is a serine protease of the duodenal brush border membrane that cleaves trypsinogen and produces active trypsin, thereby leading to the activation of many pancreatic digestive enzymes. Overlapping cDNA clones that encode the complete human enterokinase amino acid sequence were isolated from a human intestine cDNA library. Starting from the first ATG codon, the composite 3696 nt cDNA sequence contains an open reading frame of 3057 nt that encodes a 784 amino acid heavy chain followed by a 235 amino acid light chain; the two chains are linked by at least one disulfide bond. The heavy chain contains a potential N-terminal myristoylation site, a potential signal anchor sequence near the amino terminus, and six structural motifs that are found in otherwise unrelated proteins. These domains resemble motifs of the LDL receptor (two copies), complement component Clr (two copies), the metalloprotease meprin (one copy), and the macrophage scavenger receptor (one copy). The enterokinase light chain is homologous to the trypsin-like serine proteinases. These structural features are conserved among human, bovine, and porcine enterokinase. By Northern blotting, a 4.4 kb enterokinase mRNA was detected only in small intestine. The enterokinase gene was localized to human chromosome 21q21 by fluorescence in situ hybridization.

  11. Extracellular engagement of ADAM12 induces clusters of invadopodia with localized ectodomain shedding activity.

    PubMed

    Albrechtsen, Reidar; Stautz, Dorte; Sanjay, Archana; Kveiborg, Marie; Wewer, Ulla M

    2011-01-15

    Invadopodia are dynamic actin structures at the cell surface that degrade extracellular matrix and act as sites of signal transduction. The biogenesis of invadopodia, including the mechanisms regulating their formation, composition, and turnover is not entirely understood. Here, we demonstrate that antibody ligation of ADAM12, a transmembrane disintegrin and metalloprotease, resulted in the rapid accumulation of invadopodia with extracellular matrix-degrading capacity in epithelial cells expressing the αvβ3 integrin and active c-Src kinase. The induction of invadopodia clusters required an intact c-Src interaction site in the ADAM12 cytoplasmic domain, but was independent of the catalytic activity of ADAM12. Caveolin-1 and transmembrane protease MMP14/MT1-MMP were both present in the ADAM12-induced clusters of invadopodia, and cholesterol depletion prevented their formation, suggesting that lipid-raft microdomains are involved in the process. Importantly, our data demonstrate that ADAM12-mediated ectodomain shedding of epidermal growth factor receptor ligands can occur within these invadopodia. Such localized growth factor signalling offers an interesting novel biological concept highly relevant to the properties of carcinoma cells, which often show upregulated ADAM12 and β3 integrin expression, together with high levels of c-Src kinase activity.

  12. Local perturbations perturb—exponentially–locally

    SciTech Connect

    De Roeck, W. Schütz, M.

    2015-06-15

    We elaborate on the principle that for gapped quantum spin systems with local interaction, “local perturbations [in the Hamiltonian] perturb locally [the groundstate].” This principle was established by Bachmann et al. [Commun. Math. Phys. 309, 835–871 (2012)], relying on the “spectral flow technique” or “quasi-adiabatic continuation” [M. B. Hastings, Phys. Rev. B 69, 104431 (2004)] to obtain locality estimates with sub-exponential decay in the distance to the spatial support of the perturbation. We use ideas of Hamza et al. [J. Math. Phys. 50, 095213 (2009)] to obtain similarly a transformation between gapped eigenvectors and their perturbations that is local with exponential decay. This allows to improve locality bounds on the effect of perturbations on the low lying states in certain gapped models with a unique “bulk ground state” or “topological quantum order.” We also give some estimate on the exponential decay of correlations in models with impurities where some relevant correlations decay faster than one would naively infer from the global gap of the system, as one also expects in disordered systems with a localized groundstate.

  13. Aging in Sweden: local variation, local control.

    PubMed

    Davey, Adam; Malmberg, Bo; Sundström, Gerdt

    2014-08-01

    Aging in Sweden has been uniquely shaped by its history-most notably the long tradition of locally controlled services for older adults. We considered how local variations and local control shape the experience of aging in Sweden and organized the paper into 3 sections. First, we examine aging in Sweden along demography, economy, and housing. Next, we trace the origins and development of the Swedish welfare state to consider formal supports (service provision) and informal supports (caregiving and receipt of care). Finally, we direct researchers to additional data resources for understanding aging in Sweden in greater depth. Sweden was one of the first countries to experience rapid population aging. Quality of life for a majority of older Swedes is high. Local control permits a flexible and adaptive set of services and programs, where emphasis is placed on improving the quality and targeting of services that have already reached a plateau as a function of population and expenditures.

  14. Time to Go Local!

    MedlinePlus

    ... Bar Home Current Issue Past Issues Time to Go Local! Past Issues / Winter 2007 Table of Contents ... MedlinePlus.gov health topic pages, you will find "Go Local" links that take you to information about ...

  15. Temporal Non-locality

    NASA Astrophysics Data System (ADS)

    Filk, Thomas

    2013-04-01

    In this article I investigate several possibilities to define the concept of "temporal non-locality" within the standard framework of quantum theory. In particular, I analyze the notions of "temporally non-local states", "temporally non-local events" and "temporally non-local observables". The idea of temporally non-local events is already inherent in the standard formalism of quantum mechanics, and Basil Hiley recently defined an operator in order to measure the degree of such a temporal non-locality. The concept of temporally non-local states enters as soon as "clock-representing states" are introduced in the context of special and general relativity. It is discussed in which way temporally non-local measurements may find an interesting application for experiments which test temporal versions of Bell inequalities.

  16. Localized Prostate Cancer

    MedlinePlus

    ... a decision aid for men with clinically localized prostate cancer (available at http://effectivehealthcare.ahrq.gov/prostate_da) ... A Decision Aid for Men With Clinically Localized Prostate Cancer Page 1 of 24 Introduction Men with clinically ...

  17. Learning from Software Localization.

    ERIC Educational Resources Information Center

    Guo, She-Sen

    2003-01-01

    Localization is the process of adapting a product to meet the language, cultural and other requirements of a specific target environment or market. This article describes ways in which software localization impacts upon curriculum, and discusses what students will learn from software localization. (AEF)

  18. Exponential Localization of Photons

    NASA Astrophysics Data System (ADS)

    Bialynicki-Birula, Iwo

    1998-06-01

    It is shown that photons can be localized in space with an exponential falloff of the energy density and photodetection rates. The limits of localization are determined by the fundamental Paley-Wiener theorem. A direct mathematical connection between the spatial localization of photons and the decay in time of quantum mechanical systems is established.

  19. Local quantum thermal susceptibility

    PubMed Central

    De Pasquale, Antonella; Rossini, Davide; Fazio, Rosario; Giovannetti, Vittorio

    2016-01-01

    Thermodynamics relies on the possibility to describe systems composed of a large number of constituents in terms of few macroscopic variables. Its foundations are rooted into the paradigm of statistical mechanics, where thermal properties originate from averaging procedures which smoothen out local details. While undoubtedly successful, elegant and formally correct, this approach carries over an operational problem, namely determining the precision at which such variables are inferred, when technical/practical limitations restrict our capabilities to local probing. Here we introduce the local quantum thermal susceptibility, a quantifier for the best achievable accuracy for temperature estimation via local measurements. Our method relies on basic concepts of quantum estimation theory, providing an operative strategy to address the local thermal response of arbitrary quantum systems at equilibrium. At low temperatures, it highlights the local distinguishability of the ground state from the excited sub-manifolds, thus providing a method to locate quantum phase transitions. PMID:27681458

  20. Experiments with program locality.

    NASA Technical Reports Server (NTRS)

    Spirn, J. R.; Denning, P. J.

    1972-01-01

    Attempt to validate experimentally several intrinsic models for the concept of program locality. The models considered are a two-parameter simple locality model, a simple least recently used stack model, and an independent reference model. The first two models appear to produce good approximations to real world behavior, while the independent reference model, because of its static concept of locality, does very poorly. The working set is shown to be a good estimator of the simple two-parameters model's locality, provided that the locality does not change too rapidly. The working set exactly measures the locality in the case of the least recently used stack model and is thus nearly optimal for programs whose behavior can be closely approximated by this model.

  1. Local Estuary Programs

    EPA Pesticide Factsheets

    This page provides information about Local Individual Estuary Programs including links to their NEP homepages, social media, Comprehensive Conservation and Management Plans, and state of the bay reports.

  2. Comparative localization of ADAMs 10 and 15 in human cerebral cortex normal aging, Alzheimer disease and Down syndrome.

    PubMed

    Bernstein, Hans-Gert; Bukowska, Alicja; Krell, Dieter; Bogerts, Bernhard; Ansorge, Siegfried; Lendeckel, Uwe

    2003-02-01

    Using immunohistochemical techniques we studied the light microscopic localization of ADAMs (A Disintegrin And Metalloprotease) 10 and 15 in different neocortical areas of the human brain during normal aging, and also in patients with Alzheimer disease (AD) and Down syndrome (DS). ADAM 10, a putative alpha-secretase involved in Notch signaling, was found in neurons of the perinatal cortex. During aging there is an increase in intraneuronal staining intensity and in the number of cortical nerve cells that contain the enzyme. Furthermore, in AD and DS brains ADAM 10 immunoreactivity was associated with diffuse and neuritic plaques. ADAM 15 was detected in perinatal cortical pyramidal cells; during aging there was also an increase in intracellular staining and the number of stained cells per volume (cell density). In AD brains ADAM 15 was seen in a few diffuse plaques. Morphometric analysis revealed a significant reduction of ADAM 10 but not ADAM 15 immunoreactive neurons in AD brains in comparison to controls. Our findings support the idea that ADAM 10 is involved in the pathophysiology of AD and DS. ADAM 15 might be linked to AD via interaction with integrin and/or src protein tyrosine kinases.

  3. Local theory for Mott-Anderson localization

    NASA Astrophysics Data System (ADS)

    Sen, Sudeshna; Terletska, Hanna; Moreno, Juana; Vidhyadhiraja, N. S.; Jarrell, Mark

    2016-12-01

    The paramagnetic metallic phase of the Anderson-Hubbard model (AHM) is investigated using a nonperturbative local moment approach within the framework of dynamical mean-field theory with a typical medium. Our focus is on the breakdown of the metallic phase near the metal-insulators transition as seen in the single-particle spectra, scattering rates, and the associated distribution of Kondo scales. We demonstrate the emergence of a universal, underlying low-energy scale, TKpeak. This lies close to the peak of the distribution of Kondo scales obtained within the metallic phase of the paramagnetic AHM. Spectral dynamics for energies ω ≲TKpeak display Fermi liquid universality crossing over to an incoherent universal dynamics for ω ≫TKpeak in the scaling regime. Such universal dynamics indicate that within a local theory the low to moderately low-energy physics is governed by an effective, disorder renormalized Kondo screening.

  4. Localization of Interference Fringes.

    ERIC Educational Resources Information Center

    Simon, J. M.; Comastri, Silvia A.

    1980-01-01

    Discusses a proof for determining the localized fringes position arrived at when one considers the interference of two extended sources when one is able to observe fringes only at certain points in space. Shows how the localized fringes may be found in a device used to observe Newton's rings. (Author/CS)

  5. Local anesthesia: a review.

    PubMed

    Malamed, S F; Sykes, P; Kubota, Y; Matsuura, H; Lipp, M

    1992-01-01

    Local anesthetics are the most widely administered drugs in dentistry. Significant advances have been made in past decades that have greatly increased both the safety and the efficacy of these important drugs. This paper reviews the history of local anesthesia, pharmacokinetics and clinical implications, techniques, complications, and future directions in the quest for more effective pain control in dentistry.

  6. Local health traditions.

    PubMed

    Shankar, D

    1988-03-01

    A very systematic study made in Karjat, Maharasht (a tribal area in India) has found that in comparison with the official health care and primary health centers, the traditional health practices are far more comprehensive. However, although the local traditions are comprehensive in their 'scope', they nevertheless reveal several weaknesses when subjected to critical evaluation by the science of Ayurveda. For example, whereas some remedies are found to be sound, there are others that are incomplete, and some appear to be totally distorted. Similarly, the diagnostic abilities of local practitioners, while sound in some cases, are in several others found to be inadequate. As to the use of local herbs, whereas the local tradition has an amazing knowledge of local flora--the knowledge about properties of plants is in many cases incomplete. There are several reasons that may explain how and why these weaknesses have set in. 1st, the local traditions are 'oral' and in the natural course of things, oral traditions the world over have been found to decay over time. A 2nd, external, reason for the current decay of local traditions is the derision, neglect, and oppression they have suffered due to the intolerance of western scientific tradition. A 3rd reason for weaknesses in the local health stream is the breakdown of active links, during the last few centuries, with the mainstream science of Ayurveda.

  7. Interaural Coherence and Localization

    NASA Astrophysics Data System (ADS)

    Pepin, Eric

    2006-10-01

    In a study of the relationship between interaural coherence and localization ability, two experiments were performed. Both made use of a 1/3 octave band of low frequency sound and a 1/3 octave band of high frequency sound. Stimuli with coherences ranging from 0.2 to 0.8 were created in three recording environments using a KEMAR and digitally altered to eliminate interaural level differences (ILD). The environments had short, medium, and long reverberation times. The coherences were measured and were accurate to one significant figure. Experiment 1 had two goals: to determine the relationship between interaural coherence and the ability to localize using interaural time differences (ITD) and to determine if localization ability was dependent only on coherence. The relationship between coherence and localization was tested in a headphone lateralization experiment in which psychometric functions were generated. The functions revealed a linear relationship, with the ability to localize high coherence sounds breaking down quickly at small ITD. Within standard error, ITD localization appeared to be dependent only on coherence. In Experiment 2, a 3-down 1-up staircase method was employed to determine how opposing ILDs affected ITD localization. When the task could be completed, the threshold values were linearly related, however, the ability broke down at large ILDs. Both experiments provide a linear description of interaural coherence and localization, with thresholds being sharp deviations from these trends.

  8. Local Power: Tribe & Township.

    ERIC Educational Resources Information Center

    Matlala, Padi; Moloi, Dudley

    1995-01-01

    Examines the service infrastructure of a rural township in South Africa and the struggle to acquire services like water and electricity. Discusses the interaction of a system of transitional local councils and tribal authorities in the face of local government elections. (LZ)

  9. Mechanisms Regulating Protein Localization.

    PubMed

    Bauer, Nicholas C; Doetsch, Paul W; Corbett, Anita H

    2015-10-01

    Cellular functions are dictated by protein content and activity. There are numerous strategies to regulate proteins varying from modulating gene expression to post-translational modifications. One commonly used mode of regulation in eukaryotes is targeted localization. By specifically redirecting the localization of a pool of existing protein, cells can achieve rapid changes in local protein function. Eukaryotic cells have evolved elegant targeting pathways to direct proteins to the appropriate cellular location or locations. Here, we provide a general overview of these localization pathways, with a focus on nuclear and mitochondrial transport, and present a survey of the evolutionarily conserved regulatory strategies identified thus far. We end with a description of several specific examples of proteins that exploit localization as an important mode of regulation.

  10. Local Solid Shape

    PubMed Central

    Koenderink, Jan; van Doorn, Andrea

    2015-01-01

    Local solid shape applies to the surface curvature of small surface patches—essentially regions of approximately constant curvatures—of volumetric objects that are smooth volumetric regions in Euclidean 3-space. This should be distinguished from local shape in pictorial space. The difference is categorical. Although local solid shape has naturally been explored in haptics, results in vision are not forthcoming. We describe a simple experiment in which observers judge shape quality and magnitude of cinematographic presentations. Without prior training, observers readily use continuous shape index and Casorati curvature scales with reasonable resolution. PMID:27648217

  11. Local Governments Reimbursement Program

    EPA Pesticide Factsheets

    In the event of a release (or threatened release) of hazardous substances, EPA may provide a safety net of up to $25,000 per incident to local governments for expenses related to the release and associated emergency response measures.

  12. Stereotype locally convex spaces

    NASA Astrophysics Data System (ADS)

    Akbarov, S. S.

    2000-08-01

    We give complete proofs of some previously announced results in the theory of stereotype (that is, reflexive in the sense of Pontryagin duality) locally convex spaces. These spaces have important applications in topological algebra and functional analysis.

  13. Local Anesthetic Microcapsules.

    DTIC Science & Technology

    1981-04-15

    III Chemical Structure of Local Anesthetics 12 Table IV Processing Summary of Lidocaine Microencapsulation 15 Table V Lidocaine Microcapsule Size...Distribution 17 Table VI Processing Summary of Etidocaine Microencapsulation 18 Table VII Etidocaine Microcapsule Size Distribution 19 Table VIII Lidocaine...REPORT I PERIOD COVERED Annual Local Anesthetic Microcapsules 1 July 1980-30 March 1981 6. PERFORMING ORG. REPORT NUMBER 2106-1 7. AUTHOR() S

  14. Localization protected quantum order

    NASA Astrophysics Data System (ADS)

    Nandkishore, Rahul

    2015-03-01

    Many body localization occurs in isolated quantum systems, usually with strong disorder, and is marked by absence of dissipation, absence of thermal equilibration, and a memory of the initial conditions that survives in local observables for arbitrarily long times. The many body localized regime is a non-equilibrium, strongly disordered, non-self averaging regime that presents a new frontier for quantum statistical mechanics. In this talk, I point out that there exists a vast zoo of correlated many body localized states of matter, which may be classified using familiar notions of spontaneous symmetry breaking and topological order. I will point out that in the many body localized regime, spontaneous symmetry breaking can occur even at high energy densities in one dimensional systems, and topological order can occur even without a bulk gap. I will also discuss the phenomenology of imperfectly isolated many body localized systems, which are weakly coupled to a heat bath. I will conclude with a brief discussion of how these phenomena may best be detected in experiments. Collaborators: David Huse, S.L. Sondhi, Arijeet Pal, Vadim Oganesyan, A.C. Potter, Sarang Gopalakrishnan, S. Johri, R.N. Bhatt.

  15. Snake venom induced local toxicities: plant secondary metabolites as an auxiliary therapy.

    PubMed

    Santhosh, M Sebastin; Hemshekhar, M; Sunitha, K; Thushara, R M; Jnaneshwari, S; Kemparaju, K; Girish, K S

    2013-01-01

    Snakebite is a serious medical and socio-economic problem affecting the rural and agricultural laborers of tropical and sub-tropical region across the world leading to high morbidity and mortality. In most of the snakebite incidences, victims usually end up with permanent tissue damage and sequelae with high socioeconomic and psychological impacts. Although, mortality has been reduced markedly due to anti-venom regimen, it is associated with several limitations. Snake venom metalloprotease, hyaluronidase and myotoxic phospholipase A2 are the kingpins of tissue necrosis and extracellular matrix degradation. Thus, inhibition of these enzymes is considered to be the rate limiting step in the management of snakebite. Unfortunately, tissue necrosis and extracellular matrix degradation persists even after the administration of anti-venom. At present, inhibitors from snake serum and plasma, several synthetic compounds and their analogs have been demonstrated to possess anti-snake venom activities, but the use of plant metabolites for this purpose has an added advantage of traditional knowledge and will make the treatment cheaper and more accessible to the affected population. Therefore, the clinical and research forums are highly oriented towards plant metabolites and interestingly, certain phytochemicals are implicated as the antibody elicitors against venom toxicity that can be exploited in designing effective anti-venoms. Based on these facts, we have made an effort to enlist plant based secondary metabolites with antiophidian abilities and their mechanism of action against locally acting enzymes/toxins in particular. The review also describes their functional groups responsible for therapeutic beneficial and certainly oblige in designing potent inhibitors against venom toxins.

  16. Monaural Sound Localization Revisited

    NASA Technical Reports Server (NTRS)

    Wightman, Frederic L.; Kistler, Doris J.

    1997-01-01

    Research reported during the past few decades has revealed the importance for human sound localization of the so-called 'monaural spectral cues.' These cues are the result of the direction-dependent filtering of incoming sound waves accomplished by the pinnae. One point of view about how these cues are extracted places great emphasis on the spectrum of the received sound at each ear individually. This leads to the suggestion that an effective way of studying the influence of these cues is to measure the ability of listeners to localize sounds when one of their ears is plugged. Numerous studies have appeared using this monaural localization paradigm. Three experiments are described here which are intended to clarify the results of the previous monaural localization studies and provide new data on how monaural spectral cues might be processed. Virtual sound sources are used in the experiments in order to manipulate and control the stimuli independently at the two ears. Two of the experiments deal with the consequences of the incomplete monauralization that may have contaminated previous work. The results suggest that even very low sound levels in the occluded ear provide access to interaural localization cues. The presence of these cues complicates the interpretation of the results of nominally monaural localization studies. The third experiment concerns the role of prior knowledge of the source spectrum, which is required if monaural cues are to be useful. The results of this last experiment demonstrate that extraction of monaural spectral cues can be severely disrupted by trial-to-trial fluctuations in the source spectrum. The general conclusion of the experiments is that, while monaural spectral cues are important, the monaural localization paradigm may not be the most appropriate way to study their role.

  17. Representing properties locally.

    PubMed

    Solomon, K O; Barsalou, L W

    2001-09-01

    Theories of knowledge such as feature lists, semantic networks, and localist neural nets typically use a single global symbol to represent a property that occurs in multiple concepts. Thus, a global symbol represents mane across HORSE, PONY, and LION. Alternatively, perceptual theories of knowledge, as well as distributed representational systems, assume that properties take different local forms in different concepts. Thus, different local forms of mane exist for HORSE, PONY, and LION, each capturing the specific form that mane takes in its respective concept. Three experiments used the property verification task to assess whether properties are represented globally or locally (e.g., Does a PONY have mane?). If a single global form represents a property, then verifying it in any concept should increase its accessibility and speed its verification later in any other concept. Verifying mane for PONY should benefit as much from having verified mane for LION earlier as from verifying mane for HORSE. If properties are represented locally, however, verifying a property should only benefit from verifying a similar form earlier. Verifying mane for PONY should only benefit from verifying mane for HORSE, not from verifying mane for LION. Findings from three experiments strongly supported local property representation and ruled out the interpretation that object similarity was responsible (e.g., the greater overall similarity between HORSE and PONY than between LION and PONY). The findings further suggest that property representation and verification are complicated phenomena, grounded in sensory-motor simulations.

  18. Frequently Asked Questions (FAQs) About Local Foods, Local Places

    EPA Pesticide Factsheets

    Answers to frequently asked questions about EPA's Local Foods, Local Places planning assistance program to help communities revitalize downtowns, create economic opportunities, and improve access to healthy food by promoting local foods.

  19. Local Foods, Local Places 2016-2017 Application

    EPA Pesticide Factsheets

    2016-2017 application for Local Foods, Local Places, technical assistance that helps communities use local foods to create walkable and economically vibrant districts, boost economic opportunities, and improve access to healthy foods.

  20. Robust and Secure Localization

    DTIC Science & Technology

    2009-06-03

    attacker-chosen location to the nodes as they are deployed.  establish wormhole links and tunnel legitimate messages to distant locations in the...network which are beyond the radio range of the source nodes. The wormhole attack can potentially introduce significant localization error of any node

  1. Teaching Local History.

    ERIC Educational Resources Information Center

    Singer, Alan, Ed.

    2003-01-01

    This Social Science Docket theme issue focuses on teaching local history and included theme and non-themed articles, lesson plans, learning activities, and book, movie, and museum reviews designed for K-12 social studies teachers. Articles and materials in this issue are: "Editing Is Not Censorship" (Alan Singer); "Teachers Respond…

  2. Principle of relative locality

    SciTech Connect

    Amelino-Camelia, Giovanni; Freidel, Laurent; Smolin, Lee; Kowalski-Glikman, Jerzy

    2011-10-15

    We propose a deepening of the relativity principle according to which the invariant arena for nonquantum physics is a phase space rather than spacetime. Descriptions of particles propagating and interacting in spacetimes are constructed by observers, but different observers, separated from each other by translations, construct different spacetime projections from the invariant phase space. Nonetheless, all observers agree that interactions are local in the spacetime coordinates constructed by observers local to them. This framework, in which absolute locality is replaced by relative locality, results from deforming energy-momentum space, just as the passage from absolute to relative simultaneity results from deforming the linear addition of velocities. Different aspects of energy-momentum space geometry, such as its curvature, torsion and nonmetricity, are reflected in different kinds of deformations of the energy-momentum conservation laws. These are in principle all measurable by appropriate experiments. We also discuss a natural set of physical hypotheses which singles out the cases of energy-momentum space with a metric compatible connection and constant curvature.

  3. Local Area Networks.

    ERIC Educational Resources Information Center

    Bullard, David

    1983-01-01

    The proliferation of word processors, micro- and minicomputer systems, and other digital office equipment is causing major design changes in existing networks. Local Area Networks (LANs) which have adequately served terminal users in the past must now be redesigned. Implementation at Clemson is described. (MLW)

  4. Local Area Networks.

    ERIC Educational Resources Information Center

    Marks, Kenneth E.; Nielsen, Steven

    1991-01-01

    Discusses cabling that is needed in local area networks (LANs). Types of cables that may be selected are described, including twisted pair, coaxial cables (or ethernet), and fiber optics; network topologies, the manner in which the cables are laid out, are considered; and cable installation issues are discussed. (LRW)

  5. Local Area Networks.

    ERIC Educational Resources Information Center

    Nasatir, Marilyn; And Others

    1990-01-01

    Four papers discuss LANs (local area networks) and library applications: (1) "Institute for Electrical and Electronic Engineers Standards..." (Charles D. Brown); (2) "Facilities Planning for LANs..." (Gail Persky); (3) "Growing up with the Alumni Library: LAN..." (Russell Buchanan); and (4) "Implementing a LAN...at the Health Sciences Library"…

  6. Tackling a Local Problem.

    ERIC Educational Resources Information Center

    Cook, Martin

    1995-01-01

    Students studying water as a class project were concerned by levels of pollution at a nearby river and the local beach. They identified three environmental problems for research including sewage discharge, beach litter, and quality of swimming water. Research consisted of field trips which allowed for opportunities to improve skills in collecting…

  7. State and local governments

    NASA Technical Reports Server (NTRS)

    Barnes, Dennis

    1990-01-01

    The Virginia Space Grant Consortium approach to a close working relation to state and local governments is presented as a model for consideration. State government relations are especially important in that this is a primary resource in securing matching funds. Avenues for establishing these relationships are listed and discussed.

  8. Dynamic localized load balancing

    NASA Astrophysics Data System (ADS)

    Balandin, Sergey I.; Heiner, Andreas P.

    2003-08-01

    Traditionally dynamic load balancing is applied in resource-reserved connection-oriented networks with a large degree of managed control. Load balancing in connectionless networks is rather rudimentary and is either static or requires network-wide load information. This paper presents a fully automated, traffic driven dynamic load balancing mechanism that uses local load information. The proposed mechanism is easily deployed in a multi-vendor environment in which only a subset of routers supports the function. The Dynamic Localized Load Balancing (DLLB) mechanism distributes traffic based on two sets of weights. The first set is fixed and is inverse proportional to the path cost, typically the sum of reciprocal bandwidths along the path. The second weight reflects the utilization of the link to the first next hop along the path, and is therefore variable. The ratio of static weights defines the ideal load distribution, the ratio of variable weights the node-local load distribution estimate. By minimizing the difference between variable and fixed ratios the traffic distribution, with the available node-local knowledge, is optimal. The above mechanism significantly increases throughput and decreases delay from a network-wide perspective. Optionally the variable weight can include load information of nodes downstream to prevent congestion on those nodes. The latter function further improves network performance, and is easily implemented on top of the standard OSPF signaling. The mechanism does not require many node resources and can be implemented on existing router platforms.

  9. Local anesthetic update.

    PubMed Central

    MacKenzie, T. A.; Young, E. R.

    1993-01-01

    The development of new local anesthetics has not been an area of particularly active research for a number of years. However, as the use of regional anesthesia has expanded, additional anesthetic requirements and techniques have stimulated the search for newer drugs and ways of modifying existing ones. This article reviews some of the more recent developments in this field. PMID:8185087

  10. Global and Local Textuality.

    ERIC Educational Resources Information Center

    Campbell, B. G.

    Coherence and cohesion are fundamental considerations of the composing process that help to define the global and local components of texuality. Global text coherence centers on those aspects of the familiar rhetorical situation. Coherence operates at the paragraph and essay levels, answering questions about focus, tone, mode, topic, and thesis.…

  11. ADAM12 localizes with c-Src to actin-rich structures at the cell periphery and regulates Src kinase activity.

    PubMed

    Stautz, Dorte; Sanjay, Archana; Hansen, Matilde Thye; Albrechtsen, Reidar; Wewer, Ulla M; Kveiborg, Marie

    2010-01-01

    ADAM12 is an active metalloprotease playing an important role in tumour progression. Human ADAM12 exists in two splice variants: a long transmembrane form, ADAM12-L, and a secreted form, ADAM12-S. The subcellular localization of ADAM12-L is tightly regulated and involves intracellular interaction partners and signalling proteins. We demonstrate here a c-Src-dependent redistribution of ADAM12-L from perinuclear areas to actin-rich Src-positive structures at the cell periphery, and identified two separate c-Src binding sites in the cytoplasmic tail of ADAM12-L that interact with the SH3 domain of c-Src with different binding affinities. The association between ADAM12-L and c-Src is transient, but greatly stabilized when the c-Src kinase activity is disrupted. In agreement with this observation, kinase-active forms of c-Src induce ADAM12-L tyrosine phosphorylation. Interestingly, ADAM12-L was also found to enhance Src kinase activity in response to external signals, such as integrin engagement. Thus, we suggest that activated c-Src binds, phosphorylates, and redistributes ADAM12-L to specific sites at the cell periphery, which may in turn promote signalling mechanisms regulating cellular processes with importance in cancer.

  12. Broadband local dielectric spectroscopy

    NASA Astrophysics Data System (ADS)

    Labardi, M.; Lucchesi, M.; Prevosto, D.; Capaccioli, S.

    2016-05-01

    A route to extend the measurement bandwidth of local dielectric spectroscopy up to the MHz range has been devised. The method is based on a slow amplitude modulation at a frequency Ω of the excitation field oscillating at a frequency ω and the coherent detection of the modulated average electric force or force gradient at Ω. The cantilever mechanical response does not affect the measurement if Ω is well below its resonant frequency; therefore, limitations on the excitation field frequency are strongly reduced. Demonstration on a thin poly(vinyl acetate) film is provided, showing its structural relaxation spectrum on the local scale up to 45 °C higher than glass temperature, and nanoscale resolution dielectric relaxation imaging near conductive nanowires embedded in the polymer matrix was obtained up to 5 MHz frequency, with no physical reason to hinder further bandwidth extension.

  13. Local normal galaxies

    NASA Technical Reports Server (NTRS)

    Fichtel, Carl E.

    1990-01-01

    In the near future, high energy (E greater than 20 MeV) gamma ray astronomy offers the promise of a new means of examining the closest galaxies. Two and possibly three local galaxies, the Small and Large Magellanic Clouds and M31, should be visible to the high energy gamma ray telescope on the Gamma Ray Observatory, and the first should be seen by GAMMA-1. With the assumptions of adequate cosmic ray production and reasonable magnetic field strengths, both of which should likely be satisfied, specific predictions of the gamma ray emission can be made separating the concepts of the galactic and universal nature of cosmic rays. A study of the synchrotron radiation from the Large Magellanic Cloud (LMC) suggests that the cosmic ray density is similar to that in the local region of our galaxy, but not uniform. It is hoped the measurements will be able to verify this independent of assumptions about the magnetic fields in the LMC.

  14. Stable local oscillator module.

    SciTech Connect

    Brocato, Robert Wesley

    2007-11-01

    This report gives a description of the development of a Stable Local Oscillator (StaLO) multi-chip module (MCM). It is a follow-on report to SAND2006-6414, Stable Local Oscillator Microcircuit. The StaLO accepts a 100MHz input signal and produces output signals at 1.2, 3.3, and 3.6 GHz. The circuit is built as a multi-chip module (MCM), since it makes use of integrated circuit technologies in silicon and lithium niobate as well as discrete passive components. This report describes the development of an MCM-based version of the complete StaLO, fabricated on an alumina thick film hybrid substrate.

  15. THz Local Oscillator Technology

    NASA Technical Reports Server (NTRS)

    Mehdi, Imran

    2004-01-01

    The last decade has seen a number of technological advancements that have now made it possible to implement fully solid state local oscillator chains up to 2 THz. These chains are composed of cascaded planar multiplier stages that are pumped with W-band high power sources. The high power W-band sources are achieved by power combining MMIC amplifiers and can provide in access of 150 mW with about 10% bandwidth. Planar diode technology has also enabled novel circuit topologies that can take advantage of the high input power and demonstrate significant efficiencies well into the THz range. Cascaded chains to 1.9 THz have now been demonstrated with enough output power to successfully pump hot-electron bolometer mixers in this frequency range. An overview of the current State-of-the-Art of the local oscillator technology will be presented along with highlighting future trends and challenges.

  16. Local Anesthetic Microencapsulation.

    DTIC Science & Technology

    1983-11-04

    tollowing I.M. injection of microencapsulated lidocaine and etidocaine than following solution injections. Local toxicity of these microcapsule injections...Distribution 41 Table 12 Processing Summary of Lidocaine (Base) 43 Microencapsulation Table 13 Lidocaine (Base) Microcapsule Size 44 Distribution...Table 14 Processing Summary of Et’idocaine-HCl 45 Microencapsulation Table 15 Etidocaine-HCl Microcapsule Size 47 Distribution Table 16 Process Summary

  17. Local Rotational Symmetries.

    DTIC Science & Technology

    1985-08-01

    spanner wrench and the teaspoon, the pointed jaws of the wrench, and the main axes of the gourd , the pear, the squash, and the bowl of the teaspoon...regions such as the handle of the spanner wrench and the main axes of the gourd , squash, and teaspoon, and also pointed regions such as the end of the...Local Symmetry representation does not provide in- tuitively acceptable analyses for round regions, such as the lemon and the round ends of the gourd

  18. The Local Dark Matter

    SciTech Connect

    Helfer, H.L.

    2005-10-21

    The observations of the extended rotation curves of some galaxies provide important constraints upon the nature of the local dark matter present in the halos of these galaxies. Using these constraints, one can show that the halo dark matter cannot be some population of conventional astronomical objects and (most probably) cannot be a population of exotic non-interacting particles. We suggest that the halos can be regarded as large spatial fluctuations in a classic scalar field.

  19. Acoustics Local Area Network

    DTIC Science & Technology

    2013-01-31

    contract was to provide a shared computing i : resource - the acou tics local area network (ALAN) - to support ocean acoustic and related oceanographic...SECURITY CLASSIFICATION 20. UMITATION OF ABSTRACT OF REPORT: THIS PAGE OF ABSTRACT Unclassified I I ONRCtI COMPUTER V 10 11/94 STANDARD FORM 233 (REV 241) oo 0 90 " VLNV1LV HNO Og6OuLtOI, CT:tT 96/OT/0

  20. Localized solar collectors

    SciTech Connect

    Ghasemi, Hadi; Marconnet, Amy Marie; Chen, Gang; Ni, George Wei

    2016-10-04

    A localized heating structure, and method of forming same, for use in solar systems includes a thermally insulating layer having interconnected pores, a density of less than about 3000 kg/m.sup.3, and a hydrophilic surface, and an expanded carbon structure adjacent to the thermally insulating layer. The expanded carbon structure has a porosity of greater than about 80% and a hydrophilic surface.

  1. Enhanced local tomography

    DOEpatents

    Katsevich, Alexander J.; Ramm, Alexander G.

    1996-01-01

    Local tomography is enhanced to determine the location and value of a discontinuity between a first internal density of an object and a second density of a region within the object. A beam of radiation is directed in a predetermined pattern through the region of the object containing the discontinuity. Relative attenuation data of the beam is determined within the predetermined pattern having a first data component that includes attenuation data through the region. In a first method for evaluating the value of the discontinuity, the relative attenuation data is inputted to a local tomography function .function..sub..LAMBDA. to define the location S of the density discontinuity. The asymptotic behavior of .function..sub..LAMBDA. is determined in a neighborhood of S, and the value for the discontinuity is estimated from the asymptotic behavior of .function..sub..LAMBDA.. In a second method for evaluating the value of the discontinuity, a gradient value for a mollified local tomography function .gradient..function..sub..LAMBDA..epsilon. (x.sub.ij) is determined along the discontinuity; and the value of the jump of the density across the discontinuity curve (or surface) S is estimated from the gradient values.

  2. Simulating the local Universe

    NASA Astrophysics Data System (ADS)

    Sorce, J. G.

    2016-12-01

    In the local Universe, cosmic structures can be observed down to very small scales, scales on which the standard cosmological model might fail. Such detailed observations have to be compared with simulations in order to verify the predictions of different cosmological models. However, the cosmic variance can obscure the tests. More precisely, comparisons on a one-to-one basis are feasible only with simulations that look like the local Universe. Constrained by observed positions and peculiar velocities of galaxies, the simulations presented here reproduce locally the three-dimensional distribution of matter. Within a sphere of radius 100 hMpc, the observed nearby Large and Small Scale Structure is simulated with an accuracy of a few megaparsecs. These simulations include our nearest cluster neighbor, Virgo, allowing a detailed study of its formation history. It follows that the Virgo cluster has had a quiet merging history within the last seven gigayears. In the near future, zoom-in hydrodynamical simulations of the later will permit deeper comparisons with observations.

  3. Local Group Cosmology

    NASA Astrophysics Data System (ADS)

    Martínez-Delgado, David

    2013-11-01

    List of contributors; List of participants; Preface; Acknowledgments; Abbreviations; 1. The formation of the Milky Way in the CDM paradigm Ken Freeman; 2. Dark matter content and tidal effects in Local Group dwarf galaxies Steven R. Majewski; 3. Notes on the missing satellites problem James Bullock; 4. The Milky Way satellite galaxies Pavel Kroupa; 5. Stellar tidal streams Rodrigo Ibata; 6. Tutorial: the analysis of colour-magnitude diagrams David Valls-Gabaud; 7. Tutorial: modeling tidal streams using N-body simulations Jorge Peñarrubia.

  4. Local Allergic Rhinitis.

    PubMed

    Campo, Paloma; Salas, María; Blanca-López, Natalia; Rondón, Carmen

    2016-05-01

    This review focuses on local allergic rhinitis, a new phenotype of allergic rhinitis, commonly misdiagnosed as nonallergic rhinitis. It has gained attention over last decade and can affect patients from all countries, ethnic groups and ages, impairing their quality of life, and is frequently associated with conjunctivitis and asthma. Diagnosis is based on clinical history, the demonstration of a positive response to nasal allergen provocation test and/or the detection of nasal sIgE. A positive basophil activation test may support the diagnosis. Recent studies have demonstrated that allergen immunotherapy is an effective immune-modifying treatment, highlighting the importance of early diagnosis.

  5. The Cytosolic Domain of Protein-tyrosine Kinase 7 (PTK7), Generated from Sequential Cleavage by a Disintegrin and Metalloprotease 17 (ADAM17) and γ-Secretase, Enhances Cell Proliferation and Migration in Colon Cancer Cells*

    PubMed Central

    Na, Hye-Won; Shin, Won-Sik; Ludwig, Andreas; Lee, Seung-Taek

    2012-01-01

    Protein-tyrosine kinase 7 (PTK7) is a member of the defective receptor protein-tyrosine kinases and is known to function as a regulator of planar cell polarity during development. Its expression is up-regulated in some cancers including colon carcinomas. A 100-kDa fragment of PTK7 was detected in the culture media from colon cancer cells and HEK293 cells. The shed fragment was named sPTK7-Ig1–7 because its molecular mass was very similar to that of the entire extracellular domain of PTK7 that contains immunoglobulin-like loops 1 to 7 (Ig1–7). The shedding of sPTK7-Ig1–7 was enhanced by treatment with phorbol 12-myristate 13-acetate. In addition to the sPTK7-Ig1–7 found in the culture medium, two C-terminal fragments of PTK7 were detected in the cell lysates: PTK7-CTF1, which includes a transmembrane segment and a cytoplasmic domain, and PTK7-CTF2, which lacks most of the transmembrane segment from PTK7-CTF1. Analysis of PTK7 processing in the presence of various protease inhibitors or after knockdown of potential proteases suggests that shedding of PTK7 into sPTK7-Ig1–7 and PTK7-CTF1 is catalyzed by ADAM17, and further cleavage of PTK7-CTF1 into PTK7-CTF2 is mediated by the γ-secretase complex. PTK7-CTF2 localizes to the nucleus and enhances proliferation, migration, and anchorage-independent colony formation. Our findings demonstrate a novel role for PTK7 in the tumorigenesis via generation of PTK7-CTF2 by sequential cleavage of ADAM17 and γ-secretase. PMID:22665490

  6. Passive Acoustic Vessel Localization

    NASA Astrophysics Data System (ADS)

    Suwal, Pasang Sherpa

    This thesis investigates the development of a low-cost passive acoustic system for localizing moving vessels to monitor areas where human activities such as fishing, snorkeling and poaching are restricted. The system uses several off-the-shelf sensors with unsynchronized clocks where the Time Difference of Arrival (TDOA) or time delay is extracted by cross-correlation of the signal between paired sensors. The cross-correlation function uses phase correlation or Phase Transform (PHAT) which whitens the cross-spectrum in order to de-emphasize dominant frequency components. Using the locations of pairs of sensors as foci, hyperbolic equations can be defined using the time delay between them. With three or more sensors, multiple hyperbolic functions can be calculated which intersect at a unique point: the boat's location. It is also found that increasing separation distances between sensors decreased the correlation between the signals. However larger separation distances have better localization capability than with small distances. Experimental results from the Columbia and Willamette Rivers are presented to demonstrate performance.

  7. Draining the Local Void

    NASA Astrophysics Data System (ADS)

    Rizzi, Luca; Tully, R. Brent; Shaya, Edward J.; Kourkchi, Ehsan; Karachentsev, Igor D.

    2017-01-01

    Two galaxies that lie deep within the Local Void provide a test of the expectation that voids expand. The modest ({M}B∼ -14) HI bearing dwarf galaxies ALFAZOAJ1952+1428 and KK246 have been imaged with Hubble Space Telescope in order to study the stellar populations and determine distances from the luminosities of stars at the tip of the red giant branch. The mixed age systems have respective distances of 8.39 Mpc and 6.95 Mpc and inferred line-of-sight peculiar velocities of ‑114 km s‑1 and ‑66 km s‑1 toward us and away from the void center. These motions compound on the Milky Way motion of ∼230 km s‑1 away from the void. The orbits of the two galaxies are reasonably constrained by a numerical action model encompassing an extensive region that embraces the Local Void. It is unambiguously confirmed that these two void galaxies are moving away from the void center at several hundred km s‑1.

  8. Generalized local emission tomography

    DOEpatents

    Katsevich, Alexander J.

    1998-01-01

    Emission tomography enables locations and values of internal isotope density distributions to be determined from radiation emitted from the whole object. In the method for locating the values of discontinuities, the intensities of radiation emitted from either the whole object or a region of the object containing the discontinuities are inputted to a local tomography function .function..sub..LAMBDA..sup.(.PHI.) to define the location S of the isotope density discontinuity. The asymptotic behavior of .function..sub..LAMBDA..sup.(.PHI.) is determined in a neighborhood of S, and the value for the discontinuity is estimated from the asymptotic behavior of .function..sub..LAMBDA..sup.(.PHI.) knowing pointwise values of the attenuation coefficient within the object. In the method for determining the location of the discontinuity, the intensities of radiation emitted from an object are inputted to a local tomography function .function..sub..LAMBDA..sup.(.PHI.) to define the location S of the density discontinuity and the location .GAMMA. of the attenuation coefficient discontinuity. Pointwise values of the attenuation coefficient within the object need not be known in this case.

  9. Acoustic sniper localization system

    NASA Astrophysics Data System (ADS)

    Prado, Gervasio; Dhaliwal, Hardave; Martel, Philip O.

    1997-02-01

    Technologies for sniper localization have received increased attention in recent months as American forces have been deployed to various trouble spots around the world. Among the technologies considered for this task acoustics is a natural choice for various reasons. The acoustic signatures of gunshots are loud and distinctive, making them easy to detect even in high noise background environments. Acoustics provides a passive sensing technology with excellent range and non line of sight capabilities. Last but not least, an acoustic sniper location system can be built at a low cost with off the shelf components. Despite its many advantages, the performance of acoustic sensors can degrade under adverse propagation conditions. Localization accuracy, although good, is usually not accurate enough to pinpoint a sniper's location in some scenarios (for example which widow in a building or behind which tree in a grove). For these more demanding missions, the acoustic sensor can be used in conjunction with an infra red imaging system that detects the muzzle blast of the gun. The acoustic system can be used to cue the pointing system of the IR camera in the direction of the shot's source.

  10. Pattern Driven Stress Localization

    NASA Astrophysics Data System (ADS)

    Croll, Andrew; Crosby, Alfred

    2010-03-01

    The self-assembly of patterns from isotropic initial states is a major driver of modern soft-matter research. This avenue of study is directed by the desire to understand the complex physics of the varied structures found in Nature, and by technological interest in functional materials that may be derived through biomimicry. In this work we show how a simple striped phase can respond with significant complexity to an appropriately chosen perturbation. In particular, we show how a buckled elastic plate transitions into a state of stress localization using a simple, self-assembled variation in surface topography. The collection of topographic boundaries act in concert to change the state from isotropic sinusoidal wrinkles, to sharp folds or creases separated by relatively flat regions. By varying the size of the imposed topographic pattern or the wavelength of the wrinkles, we construct a state diagram of the system. The localized state has implications for both biological systems, and for the control of non-linear pattern formation.

  11. Locality and reality

    SciTech Connect

    Stapp, Henry P.

    1980-10-01

    Einstein's principle that no signal travels faster than suggests that observations in one spacetime region should not depend on whether or not a radioactive decay is detected in a spacelike separated region. This locality property is incompatible with the predictions of quantum theory, and this incompatibility holds independently of the questions of realism, objective reality, and hidden variables. It holds both in the pragmatic quantum theory of Bohr and in realistic frameworks. It is shown here to hold in a completed realistic quantum theory that reconciles Einstein's demand for a description of reality itself with Bohr's contention that quantum theory is complete. This completed realistic quantum theory has no hidden variables, and no objective reality in which observable attributes can become definite independently of observers. The, theory is described in some detail, with particular attention to those aspects related to the question of locality. This completed realistic quantum theory is in principle more comprehensive than Bohr.' s pragmatic quantum theory because it is not limited in principle by the requirement that the observed system be physically separated from the observing one. Applications are discussed.

  12. A Global Census of Fission Yeast Deubiquitinating Enzyme Localization and Interaction Networks Reveals Distinct Compartmentalization Profiles and Overlapping Functions in Endocytosis and Polarity

    PubMed Central

    Kouranti, Ilektra; McLean, Janel R.; Feoktistova, Anna; Liang, Ping; Johnson, Alyssa E.; Roberts-Galbraith, Rachel H.; Gould, Kathleen L.

    2010-01-01

    Ubiquitination and deubiquitination are reciprocal processes that tune protein stability, function, and/or localization. The removal of ubiquitin and remodeling of ubiquitin chains is catalyzed by deubiquitinating enzymes (DUBs), which are cysteine proteases or metalloproteases. Although ubiquitination has been extensively studied for decades, the complexity of cellular roles for deubiquitinating enzymes has only recently been explored, and there are still several gaps in our understanding of when, where, and how these enzymes function to modulate the fate of polypeptides. To address these questions we performed a systematic analysis of the 20 Schizosaccharomyces pombe DUBs using confocal microscopy, proteomics, and enzymatic activity assays. Our results reveal that S. pombe DUBs are present in almost all cell compartments, and the majority are part of stable protein complexes essential for their function. Interestingly, DUB partners identified by our study include the homolog of a putative tumor suppressor gene not previously linked to the ubiquitin pathway, and two conserved tryptophan-aspartate (WD) repeat proteins that regulate Ubp9, a DUB that we show participates in endocytosis, actin dynamics, and cell polarity. In order to understand how DUB activity affects these processes we constructed multiple DUB mutants and find that a quintuple deletion of ubp4 ubp5 ubp9 ubp15 sst2/amsh displays severe growth, polarity, and endocytosis defects. This mutant allowed the identification of two common substrates for five cytoplasmic DUBs. Through these studies, a common regulatory theme emerged in which DUB localization and/or activity is modulated by interacting partners. Despite apparently distinct cytoplasmic localization patterns, several DUBs cooperate in regulating endocytosis and cell polarity. These studies provide a framework for dissecting DUB signaling pathways in S. pombe and may shed light on DUB functions in metazoans. PMID:20838651

  13. All Holdings Are Local: Archivists and Local Government Archives

    ERIC Educational Resources Information Center

    James, Russell D.

    2004-01-01

    Archivists working in repositories that contain local government records play an integral role within the local community. Unlike their colleagues in state, regional, or national repositories, the local government archivist concentrates on a small geographic region and demonstrates knowledge of the politics, history, and socio-economics of the…

  14. THz Local Oscillator Sources

    NASA Astrophysics Data System (ADS)

    Mehdi, Imran; Schlecht, Erich; Chattopadhyay, Goutam; Siegel, Peter H.

    Most operational Submillimeter-wave radio telescopes, both space borne and ground based, employ local oscillator sources based on Gunn diodes followed by whisker contacted Schottky multipliers. Enough progress, however, has been made on a number of fronts to conclude that next generation of radio telescopes that become operational in the new Millennium will have a different local oscillator (LO) generation architecture. MMIC power amplifiers with impressive gain in the Ka- to-W band have enabled the use of microwave synthesizers which can then be actively multiplied to provide a frequency agile power source beyond 100 GHz. This medium power millimeter source can then be amplified to enable efficient pumping of follow-on balanced multiplier stages. Input power to the multipliers can be further enhanced by power combining to achieve close to half a Watt at W-band. An 800 GHz three-stage multiplier chain, implemented this way has demonstrated a peak output power of 1 mW. A second advance in LO generation lies in the Schottky diode varactor technology. Planar Schottky diode multipliers have now been demonstrated up to 1500 GHz and it can be assumed that most of the future multiplier chains will be based on these robust devices rather than the whisker contacted diode of the past. The ability to produce planar GaAs diode chips deep into the THz range, with submicron dimensions, has opened up a wide range of circuit design space which can be taken advantage of to improve efficiency, bandwidth, and power handling capability of the multipliers. A third breakthrough has been the demonstration of photonic based LO sources utilizing GaAs photomixers. These sources, though not yet implemented in robust space borne missions, offer a number of advantages over their electronic counterparts, including extremely broad tuning, fiber coupled components, and solid-state implementation. Another development, which holds some promise, is the use of micro-machining technology to implement

  15. Stable local oscillator microcircuit.

    SciTech Connect

    Brocato, Robert Wesley

    2006-10-01

    This report gives a description of the development of a Stable Local Oscillator (StaLO) Microcircuit. The StaLO accepts a 100MHz input signal and produces output signals at 1.2, 3.3, and 3.6 GHz. The circuit is built as a multi-chip module (MCM), since it makes use of integrated circuit technologies in silicon and lithium niobate as well as discrete passive components. The StaLO uses a comb generator followed by surface acoustic wave (SAW) filters. The comb generator creates a set of harmonic components of the 100MHz input signal. The SAW filters are narrow bandpass filters that are used to select the desired component and reject all others. The resulting circuit has very low sideband power levels and low phase noise (both less than -40dBc) that is limited primarily by the phase noise level of the input signal.

  16. Headphone localization of speech

    NASA Technical Reports Server (NTRS)

    Begault, Durand R.; Wenzel, Elizabeth M.

    1993-01-01

    Three-dimensional acoustic display systems have recently been developed that synthesize virtual sound sources over headphones based on filtering by head-related transfer functions (HRTFs), the direction-dependent spectral changes caused primarily by the pinnae. In this study, 11 inexperienced subjects judged the apparent spatial location of headphone-presented speech stimuli filtered with nonindividualized HRTFs. About half of the subjects 'pulled' their judgments toward either the median or the lateral-vertical planes, and estimates were almost always elevated. Individual differences were pronounced for the distance judgments; 15 to 46 percent of stimuli were heard inside the head, with the shortest estimates near the median plane. The results suggest that most listeners can obtain useful azimuth information from speech stimuli filtered by nonindividualized HRTFs. Measurements of localization error and reversal rates are comparable with a previous study that used broadband noise stimuli.

  17. Local control stations

    SciTech Connect

    Brown, W.S.; Higgins, J.C.; Wachtel, J.A.

    1993-05-01

    This paper describes research concerning the effects of human engineering design at local control stations (i.e., operator interfaces located outside the control room) on human performance and plant safety. The research considered both multifunction panels (e.g. remote shutdown panels) as well as single-function interfaces (e.g., valves, breakers, gauges, etc.). Changes in performance shaping factors associated with variations in human engineering at LCSs were estimated based on expert opinion. By means of a scaling procedure, these estimates were used to modify the human error probabilities in a PRA model, which was then employed to generate estimates of plant risk and scoping-level value/impact ratios for various human engineering upgrades. Recent documentation of human engineering deficiencies at single-function LCSs was also reviewed, and an assessment of the current status of LCSs with respect to human engineering was conducted.

  18. Headphone localization of speech.

    PubMed

    Begault, D R; Wenzel, E M

    1993-06-01

    Three-dimensional acoustic display systems have recently been developed that synthesize virtual sound sources over headphones based on filtering by head-related transfer functions (HRTFs), the direction-dependent spectral changes caused primarily by the pinnae. In this study 11 inexperienced subjects judged the apparent spatial location of headphone-presented speech stimuli filtered with non-individualized HRTFs. About half of the subjects "pulled" their judgments toward either the median or the lateral-vertical planes, and estimates were almost always elevated. Individual differences were pronounced for the distance judgments; 15% to 46% of stimuli were heard inside the head, with the shortest estimates near the median plane. The results suggest that most listeners can obtain useful azimuth information from speech stimuli filtered by nonindividualized HRTFs. Measurements of localization error and reversal rates are comparable with a previous study that used broadband noise stimuli.

  19. Localized wave pulse experiments

    SciTech Connect

    Chambers, D L; Henderson, T L; Krueger, K L; Lewis, D K; Zilkowski, R N

    1999-06-01

    The Localized Wave project of the Strategic System Support Program has recently finished an experiment in cooperation with the Advanced SONAR group of the Applied Research Laboratory of the University of Texas at Austin. The purpose of the experiment was three-fold. They wanted to see if (1) the LW pulse could propagate over significant distances, to see if (2) a new type of array and drive system specifically designed for the pulse would increase efficiency over single frequency tone bursts, and to see if (3) the complexity of our 24 channel drivers resulted in better efficiency than a single equivalent pulse driving a piston. In the experiment, several LW pulses were launched from the Lake Travis facility and propagated over distances of either 100 feet or 600 feet, through a thermocline for the 600 foot measurements. The results show conclusively that the Localized Wave will propagate past the near field distance. The LW pulses resulted in extremely broad frequency band width pulses with narrow spatial beam patterns and unmeasurable side lobes. Their array gain was better than most tone bursts and further, were better than their equivalent piston pulses. This marks the first test of several Low Diffraction beams against their equivalent piston pulses, as well as the first propagation of LW pulses over appreciable distances. The LW pulse is now proven a useful tool in open water, rather than a laboratory curiosity. The experimental system and array were built by ARL, and the experiments were conducted by ARL staff on their standard test range. The 600 feet measurements were made at the farthest extent of that range.

  20. Estimation of local spatial scale

    NASA Technical Reports Server (NTRS)

    Watson, Andrew B.

    1987-01-01

    The concept of local scale asserts that for a given class of psychophysical measurements, performance at any two visual field locations is equated by magnifying the targets by the local scale associated with each location. Local scale has been hypothesized to be equal to cortical magnification or alternatively to the linear density of receptors or ganglion cells. Here, it is shown that it is possible to estimate local scale without prior knowledge about the scale or its physiological basis.

  1. Federal, State, and Local regulations

    SciTech Connect

    Kelly, J.M.; Brandenburg, B.L. )

    1991-08-01

    This article is a review of federal, state, and local regulations pertinent treatment of leachate from hazardous materials landfills in California. The topics covered include under federal regulations: pretreatment, whole-effluent toxicity, hazardous waste regulation; under state regulations: hazardous waste regulations, air toxics, environmental quality act; under local regulations: local limits, toxicity-regional water quality board, air emissions and district code.

  2. Local Government: The Learning Laboratory.

    ERIC Educational Resources Information Center

    Degelman, Charles, Ed.

    2001-01-01

    This issue of "Service-Learning Network" looks at the ways that service learning can transform local government into a learning laboratory for civic education. The first article, "Creating the Missing Link: Local Government, Service Learning, and Civic Education" (Todd Clark), introduces the issue. "Service Learning and Local Government" (Ann…

  3. Localization algorithm and error analysis for micro radio-localizer

    NASA Astrophysics Data System (ADS)

    Li, Xudong; Wang, Xiaohao; Li, Qiang; Zhao, Huijie

    2006-11-01

    After more than ten years' research efforts on the Micro Aerial Vehicle (MAV) since it was proposed in 1990s, the stable flying platform has been matured. The next reasonable goal is to implement more practical applications for MAVs. Equipped with a micro radio-localizer, MAVs have the ability of localizing a target that transmitting radio signals, and further can be a novel promising Anti-Radiation device. A micro radio-localizer prototype and its localization principle and localization algorithm are proposed. The error analysis of the algorithm is also discussed. On the basis of the comparison of the often-used radio localization method, considering the MAVs' inherent limitation on the dimension of the antennas, a signal intensity and guidance information based localization method is proposed. Under the assumption that the electromagnetic wave obeys the free-space spreading model and the signal's power keeps unchanged, the measuring equations under different target motions are established. Localization algorithm is derived. The determination of several factors such as the number of measuring positions, numerical solving method and initial solution is discussed. Error analysis of the localization algorithm is also proposed by utilizing error analysis theory. A radio-localizer prototype is developed and experiment results are shown as well.

  4. Local Universe Dwarf Galaxies

    NASA Astrophysics Data System (ADS)

    Carignan, Claude

    2015-08-01

    One of the outstanding problems in cosmology is addressing the "small-scale crisis" and understanding structure formation at the smallest scales. Standard Lambda Cold Dark Matter cosmological simulations of Milky Way-size DM halos predict many more DM sub-halos than the number of dwarf galaxies observed. This is the so-called Missing Satellites Problem. The most popular interpretation of the Missing Satellites Problem is that the smallest dark matter halos in the universe are extremely inefficient at forming stars. The virialized extent of the Milky Way's halo should contain ~500 satellites, while only ˜100 satellites and dwarfs are observed in the whole Local Group. Despite the large amount of theoretical work and new optical observations, the discrepancy, even if reduced, still persists between observations and hierarchical models, regardless of the model parameters. It may be possible to find those isolated ultra-faint missing dwarf galaxies via their neutral gas component, which is one of the goals we are pursuing with the SKA precursor KAT-7 in South Africa, and soon with the SKA pathfinder MeerKAT.

  5. Local slope stability analysis

    NASA Astrophysics Data System (ADS)

    Hattendorf, I.; Hergarten, St.; Neugebauer, H. J.

    Mass movements under the influence of gravity occur as result of diverse disturbing and destabilizing processes, for example of climatic or anthropological origin. The stability of slopes is mainly determined by the geometry of the land-surface and designated slip-horizon. Further contributions are supplied by the pore water pressure, cohesion and friction. All relevant factors have to be integrated in a slope stability model, either by measurements and estimations (like phenomenological laws) or derived from physical equations. As result of stability calculations, it's suitable to introduce an expectation value, the factor-of-safety, for the slip-risk. Here, we present a model based on coupled physical equations to simulate hardly measurable phenomenons, like lateral forces and fluid flow. For the displacements of the soil-matrix we use a modified poroelasticity-equation with a Biot-coupling (Biot 1941) for the water pressure. Latter is described by a generalized Boussinesq equation for saturated-unsaturated porous media (Blendinger 1998). One aim of the calculations is to improve the knowledge about stability-distributions and their temporal variations. This requires the introduction of a local factor-of-safety which is the main difference to common stability models with global stability estimations. The reduction of immediate danger is still the emergent task of the most slope and landslide investigations, but this model is also useful with respect to understand the governing processes of landform evolution.

  6. Local reduction in physics

    NASA Astrophysics Data System (ADS)

    Rosaler, Joshua

    2015-05-01

    A conventional wisdom about the progress of physics holds that successive theories wholly encompass the domains of their predecessors through a process that is often called "reduction." While certain influential accounts of inter-theory reduction in physics take reduction to require a single "global" derivation of one theory's laws from those of another, I show that global reductions are not available in all cases where the conventional wisdom requires reduction to hold. However, I argue that a weaker "local" form of reduction, which defines reduction between theories in terms of a more fundamental notion of reduction between models of a single fixed system, is available in such cases and moreover suffices to uphold the conventional wisdom. To illustrate the sort of fixed-system, inter-model reduction that grounds inter-theoretic reduction on this picture, I specialize to a particular class of cases in which both models are dynamical systems. I show that reduction in these cases is underwritten by a mathematical relationship that follows a certain liberalized construal of Nagel/Schaffner reduction, and support this claim with several examples. Moreover, I show that this broadly Nagelian analysis of inter-model reduction encompasses several cases that are sometimes cited as instances of the "physicist's" limit-based notion of reduction.

  7. Local positioning system

    SciTech Connect

    Kyker, R.

    1995-07-25

    Navigation systems have been vital to transportation ever since man took to the air and sea. Early navigation systems utilized the sextant to navigate by starlight as well as the magnetic needle compass. As electronics and communication technologies improved, inertial navigation systems were developed for use in ships and missile delivery. These systems consisted of electronic compasses, gyro-compasses, accelerometers, and various other sensors. Recently, systems such as LORAN and the Global Positioning System (GPS) have utilized the properties of radio wave propagation to triangulate position. The Local Positioning System (LPS), described in this paper, is an implementation of a limited inertial navigation system designed to be used on a bicycle. LPS displays a cyclist`s current position relative to a starting location. This information is displayed in Cartesian-like coordinates. To accomplish this, LPS relies upon two sensors, an electronic compass sensor and a distance sensor. The compass sensor provides directional information while the distance sensor provides the distance traveled. This information yields a distance vector for each point in time which when summed produces the cyclist`s current position. LPS is microprocessor controlled and is designed for a range of less than 90 miles.

  8. Impact of Local Sensors

    NASA Technical Reports Server (NTRS)

    Watson, Leela R.; Bauman, William H., III

    2008-01-01

    Forecasters at the 45th Weather Squadron (45 WS) use observations from the Kennedy Space Center (KSC) and Cape Canaveral Air Force Station (CCAFS) wind tower network and the CCAFS (XMR) daily rawinsonde observations (RAOB) to issue and verify wind advisories and warnings for operations. These observations are also used by the National Weather Service (NWS) Spaceflight Meteorology Group (SMG) in Houston, Texas and the NWS Melbourne, Florida (NWS MLB) to initialize their locally-run mesoscale models. In addition, SMG uses these observations to support shuttle landings at the Shuttle Landing Facility (SLF). Due to impending budget cuts, some or all of the KSC/CCAFS wind towers on the east-central Florida mainland and the XMR RAOBs may be eliminated. The locations of the mainland towers and XMR RAOB site are shown in Figure I. The loss of these data may impact the forecast capability of the 45 WS, SMG and NWS MLB. The AMU was tasked to conduct an objective independent modeling study to help determine how important these observations are to the accuracy of the model output used by the forecasters. To accomplish this, the Applied Meteorology Unit (AMU) performed a sensitivity study using the Weather Research and Forecasting (WRF) model initialized with and without KSC/CCAFS wind tower and XMR RAOB data.

  9. Transcontinental local area network

    NASA Astrophysics Data System (ADS)

    Hunt, Gareth

    2000-06-01

    The National Radio Astronomy Observatory (NRAO) has facilities at 17 different locations scattered throughout the USA. These vary in size from the major laboratories occupied by research and support staff to the ten individual antennas of the Very Long Baseline Array. As is typical in astronomy, many sites are in remote locations, which are not well served with modern communication capabilities. Until 1996, the NRAO's internal network was achieved via the Internet; most sites simply had a local port to the Internet and the traffic was routed tortuously to the other locations. The burgeoning demand for Internet bandwidth was (and still is) growing faster than the services could be enhanced, and this led to intolerably slow response times and unacceptably low achieved data rates. To solve this problem, the NRAO acquired a frame relay intranet from AT&T to connect ten of its locations. The operating cost is approximately the same as the multiple Internet connections, but with vastly improved throughput and reliability. Recently, the access to the four major sites has been upgraded to support video conferencing.

  10. Local load shedding

    SciTech Connect

    Adibi, M.M.; Thorne, D.K. )

    1988-08-01

    Equipment overloads in an underground transmission network are caused by unscheduled outages. Repairs or replacements of damaged cables and/or transformers in urban areas are inherently difficult and time-consuming. Therefore, for overloads greatly in excess of short-time ratings, speed of load shedding is of paramount importance. Under such conditions, the system operator is faced with: recognizing the problem, determining the course of action and shedding the correct amount of load at the right locations. These tasks are difficult to perform, particularly under pressure of time. Reliance on pre-specified load shedding lists is not satisfactory since the load shedding lists do not necessarily match the amounts and locations of the required loads to be shed. Clearly, there has been a need for a local load shedding scheme, which in the first order of importance, would relieve the overloaded equipment within the time limits imposed by the equipment short-time ratings and in the second order of importance, would ''minimize'' the amount of load to be curtailed. This paper describes an approach which meets the dual objective, providing a practical solution to a difficult engineering/operating problem.

  11. All strategy is local.

    PubMed

    Greenwald, Bruce; Kahn, Judd

    2005-09-01

    The aim of strategy is to master a market environment by understanding and anticipating the actions of other economic agents, especially competitors. A firm that has some sort of competitive advantage-privileged access to customers, for instance--will have relatively few competitors to contend with, since potential competitors without an advantage, if they have their wits about them, will stay away. Thus, competitive advantages are actually barriers to entry and vice versa. In markets that are exposed, by contrast, competition is intense. If the incumbents have even brief success in earning greater than normal returns on investments, new entrants will swarm in to grab a share of the profits. Sooner or later, the additional competition will push returns as far down as the firms' costs of capital. For firms operating in such markets, the only choice is to forget about strategy and run the business as efficiently as possible. Barriers to entry are easier to maintain in a competitive arena that is "local", either in the geographic sense or in the sense of being limited to one product or a handful of related ones. The two most powerful competitive advantages-customer captivity and economies of scale-are more achievable and sustainable in circumscribed markets of this kind. Their opposites are the open markets and host of rivals that are features of globalization. Compapies entering such markets risk frittering away the advantages they secured on smaller playing fields.., Ifa company wants to grow but still obtain superior returns, the authors argue, the best strategy is to dominate a series of discrete but preferably contiguous markets and then expand only at their edges. WalMart's diminishing margins over the past 15 years are strong evidence of the danger of proceeding otherwise.

  12. A Local Galilean Invariant Thermostat.

    PubMed

    Groot, Robert D

    2006-05-01

    The thermostat introduced recently by Stoyanov and Groot (J. Chem. Phys. 2005, 122, 114112) is analyzed for inhomogeneous systems. This thermostat has one global feature, because the mean temperature used to drive the system toward equilibrium is a global average. The consequence is that the thermostat locally conserves energy rather than temperature. Thus, local temperature variations can be long-lived, although they do average out by thermal diffusion. To obtain a faster local temperature equilibration, a truly local thermostat must be introduced. To conserve momentum and, hence, to simulate hydrodynamic interactions, the thermostat must be Galilean invariant. Such a local Galilean invariant thermostat is studied here. It is shown that, by defining a local temperature on each particle, the ensemble is locally isothermal. The local temperature is obtained from a local square velocity average around each particle. Simulations on the ideal gas show that this local Nosé-Hoover algorithm has a similar artifact as dissipative particle dynamics:  the ideal gas pair correlation function is slightly distorted. This is attributed to the fact that the thermostat compensates fluctuations that are natural within a small cluster of particles. When the cutoff range rc for the square velocity average is increased, systematic errors decrease proportionally to rc(-)(3/2); hence, the systematic error can be made arbitrary small.

  13. Performing a local barrier operation

    DOEpatents

    Archer, Charles J; Blocksome, Michael A; Ratterman, Joseph D; Smith, Brian E

    2014-03-04

    Performing a local barrier operation with parallel tasks executing on a compute node including, for each task: retrieving a present value of a counter; calculating, in dependence upon the present value of the counter and a total number of tasks performing the local barrier operation, a base value, the base value representing the counter's value prior to any task joining the local barrier; calculating, in dependence upon the base value and the total number of tasks performing the local barrier operation, a target value of the counter, the target value representing the counter's value when all tasks have joined the local barrier; joining the local barrier, including atomically incrementing the value of the counter; and repetitively, until the present value of the counter is no less than the target value of the counter: retrieving the present value of the counter and determining whether the present value equals the target value.

  14. Performing a local barrier operation

    DOEpatents

    Archer, Charles J; Blocksome, Michael A; Ratterman, Joseph D; Smith, Brian E

    2014-03-04

    Performing a local barrier operation with parallel tasks executing on a compute node including, for each task: retrieving a present value of a counter; calculating, in dependence upon the present value of the counter and a total number of tasks performing the local barrier operation, a base value of the counter, the base value representing the counter's value prior to any task joining the local barrier; calculating, in dependence upon the base value and the total number of tasks performing the local barrier operation, a target value, the target value representing the counter's value when all tasks have joined the local barrier; joining the local barrier, including atomically incrementing the value of the counter; and repetitively, until the present value of the counter is no less than the target value of the counter: retrieving the present value of the counter and determining whether the present value equals the target value.

  15. Global/Local Dynamic Models

    SciTech Connect

    Pfeffer, A; Das, S; Lawless, D; Ng, B

    2006-10-10

    Many dynamic systems involve a number of entities that are largely independent of each other but interact with each other via a subset of state variables. We present global/local dynamic models (GLDMs) to capture these kinds of systems. In a GLDM, the state of an entity is decomposed into a globally influenced state that depends on other entities, and a locally influenced state that depends only on the entity itself. We present an inference algorithm for GLDMs called global/local particle filtering, that introduces the principle of reasoning globally about global dynamics and locally about local dynamics. We have applied GLDMs to an asymmetric urban warfare environment, in which enemy units form teams to attack important targets, and the task is to detect such teams as they form. Experimental results for this application show that global/local particle filtering outperforms ordinary particle filtering and factored particle filtering.

  16. Localized structure of Euglena bioconvection

    NASA Astrophysics Data System (ADS)

    Iima, Makoto; Shoji, Erika; Awazu, Akinori; Nishimori, Hiraku; Izumi, Shunsuke; Hiroshima University Collaboration

    2013-11-01

    Bioconvection of a suspension of Euglena gracilis, a photosensitive flagellate whose body length is approximately 50 micrometers, was experimentally studied. Under strong light intensity, Euglena has a negative phototaxis; they tend to go away from the light source. When the bright illumination is given from the bottom, a large scale spatio-temporal pattern is generated as a result of interaction between Euglena and surrounding flow. Recently, localized convection pattern had been reported, however, the generation process and interaction of the localized convection cells has not been analyzed. We performed experimental study to understand the localization mechanism, in particular, the onset of bioconvection and lateral localization behavior due to phototaxis. Experiments started from different initial condition suggests a bistability near the onset of the convection as binary fluid convection that also shows localized convection cells. Dynamics of localized convections cells, which is similar to the binary fluid convection case although the basic equations are not the same, is also reported.

  17. Auditory Spatial Perception: Auditory Localization

    DTIC Science & Technology

    2012-05-01

    Teas , D. C.; Jeffress, L. A. Localization of High Frequency Tones. Journal of the Acoustical Society of America 1957, 29, 988–991. Feinstein, S...Neurophysiology 2001, 86, 2647–2666. Itoh, M.; Adel, B. von; Kelly, J. B. Sound Localization after Transection of the Commissure of Probst in the Albino Rat...Neurology 1957, 7, 655–663. Sandel, T. T.; Teas , D. C.; Feddersen, W. E.; Jeffress, L. A. Localization of Sound From Single and Paired Sources. Journal

  18. Localization oscillation in antidot lattices

    NASA Astrophysics Data System (ADS)

    Uryu, S.; Ando, T.

    1998-06-01

    The Anderson localization in square and hexagonal antidot lattices is numerically studied with the use of a Thouless number method. It is revealed that localization is very sensitive to the aspect ratio between the antidot diameter and the lattice constant. In a hexagonal lattice, both the Thouless number and the localization length oscillate with the period equal to the Al’tshuler-Aronov-Spivak oscillation. The oscillation is quite weak in a square lattice.

  19. Adverse responses to local anaesthetics.

    PubMed

    Fisher, M M; Graham, R

    1984-11-01

    Progressive challenge was used to investigate twenty-seven patients with a history of an adverse response to local anaesthesia. True allergy was detected in only one patient. The method does not exclude reactions to additives and preservatives in local anaesthetics. If preservative-free local anaesthetics are used for subsequent exposure in patients with no response to progressive challenge, subsequent exposure is safe. The possibility that some of these patients may be reacting to preservatives in the solutions cannot be excluded by such testing. Where possible preservative-free local anaesthetic preparations should be used for subsequent anaesthesia.

  20. Alleged allergy to local anaesthetics.

    PubMed

    Fisher, M M; Bowey, C J

    1997-12-01

    The aim of this study was to determine the incidence of true local anaesthetic allergy in patients with an alleged history of local anaesthetic allergy and whether subsequent exposure to local anaesthetics is safe. Two hundred and eight patients with a history of allergy to local anaesthesia were referred over a twenty-year period to our Anaesthetic Allergy Clinic. In this open study, intradermal testing was performed in three patients and progressive challenge in 202 patients. Four patients had immediate allergy and four patients delayed allergic reactions. One hundred and ninety-seven patients were not allergic to local anaesthetics. In 39 patients an adverse response to additives in local anaesthetic solutions could not be excluded. In all but one patient local anaesthesia has been given uneventfully subsequently. A history of allergy to local anaesthesia is unlikely to be genuine and local anaesthetic allergy is rare. In most instances LA allergy can be excluded from the history and the safety of LA verified by progressive challenge.

  1. Indoor localization using passive RFID

    NASA Astrophysics Data System (ADS)

    Vastianos, George E.; Kyriazanos, Dimitris M.; Segou, Olga E.; Mitilineos, Stelios A.; Thomopoulos, Stelios C. A.

    2011-06-01

    Radio frequency identification (RFID) systems based on passive tags are used successfully in a wide range of object identification applications. However, the increasing needs to meet new demands on applications of localization and tracking create a new field for evolution of the RFID technology. This paper presents the design, implementation, and evaluation of a cost-effective localization system for in-building usage that is able to localize objects that carry passive RFID tags. The RFID reading is performed by a single Reader and an array of directional antennas through multiplexing. Evaluation and experimental results from three localization algorithms based on RSSI are presented.

  2. Empowering an Indigenous Rural Community: Local Teachers for Local Schools.

    ERIC Educational Resources Information Center

    Delany, John; Wenmoth, Derek

    2001-01-01

    Christchurch College of Education (New Zealand) offers a distance teacher education course to a rural Maori community experiencing a teacher shortage. Program success is attributed to local initiative in establishing the program, significant local input into program development, attention to the student profile in course design and delivery,…

  3. Agriculture and Locality Interrelationships: Perspectives of Local Officials and Farmers.

    ERIC Educational Resources Information Center

    Moxley, Robert L.; Liles, James

    The focus of this study is the impact of urban activities (such as industry) and local governments on agriculture and the impact of agriculture on localities. This report is based on a qualitative study of an agricultural county, and interviews with community and county officials and farmers. The perceptions and opinions of officials are compared…

  4. Local State and Sector Theory in Local Quantum Physics

    NASA Astrophysics Data System (ADS)

    Ojima, Izumi; Okamura, Kazuya; Saigo, Hayato

    2016-06-01

    We define a new concept of local states in the framework of algebraic quantum field theory (AQFT). Local states are a natural generalization of states and give a clear vision of localization in the context of QFT. In terms of them, we can find a condition from which follows automatically the famous DHR selection criterion in DHR-DR theory. As a result, we can understand the condition as consequences of physically natural state preparations in vacuum backgrounds. Furthermore, a theory of orthogonal decomposition of completely positive (CP) maps is developed. It unifies a theory of orthogonal decomposition of states and order structure theory of CP maps. Using it, localized version of sectors is formulated, which gives sector theory for local states with respect to general reference representations.

  5. [Matrix metalloproteases as molecular markers in gastric cancer].

    PubMed

    de la Peña, Sol; Sampieri, Clara L; León-Córdoba, Kenneth

    2010-02-06

    Gastric cancer is the second leading cause of cancer-associated mortality in the world. Prognosis in patients with gastric cancer is difficult to establish because it is commonly diagnosed when gastric wall invasion and metastasis have occurred. Currently, some members of the extracellular matrix metalloproteinases have been identified, whose expression in gastric tumor tissue is significantly elevated compared to healthy gastric tissue. Matrix metalloproteinases are 24 zinc-dependent endopeptidases that catalyze the proteolysis of the extracellular matrix. This degradation allows the cancer cells invade the surrounding stroma and trigger metastasis. Upregulation of certain matrix metalloproteinases in gastric cancer has been associated with a poor prognosis and elevated invasive capacity. This review compiles evidence about the genetic expression of matrix metalloproteinases in gastric cancer and their role in tumour invasion and metastasis, emphasizing their potential as molecular markers of prognosis.

  6. Endothelin-converting enzymes and related metalloproteases in Alzheimer's disease.

    PubMed

    Pacheco-Quinto, Javier; Herdt, Aimee; Eckman, Christopher B; Eckman, Elizabeth A

    2013-01-01

    The efficient clearance of amyloid-β (Aβ) is essential to modulate levels of the peptide in the brain and to prevent it from accumulating in senile plaques, a hallmark of Alzheimer's disease (AD) pathology.We and others have shown that failure in Aβ catabolism can produce elevations in Aβ concentration similar to those observed in familial forms of AD. Based on the available evidence, it remains plausible that in late-onset AD, disturbances in the activity of Aβ degrading enzymes could induce Aβ accumulation, and that this increase could result in AD pathology. The following review presents a historical perspective of the parallel discovery of three vasopeptidases (neprilysin and endothelin-converting enzymes-1 and -2) as important Aβ degrading enzymes. The recognition of the role of these vasopeptidases in Aβ degradation, beyond bringing to light a possible explanation of how cardiovascular risk factors may influence AD risk, highlights a possible risk of the use of inhibitors of these enzymes for other clinical indications such as hypertension. We will discuss in detail the experiments conducted to assess the impact of vasopeptidase deficiency (through pharmacological inhibition or genetic mutation) on Aβ accumulation, as well as the cooperative effect of multiple Aβ degrading enzymes to regulate the concentration of the peptide at multiple sites, both intracellular and extracellular, throughout the brain.

  7. Randomness and Non-Locality

    NASA Astrophysics Data System (ADS)

    Senno, Gabriel; Bendersky, Ariel; Figueira, Santiago

    2016-07-01

    The concepts of randomness and non-locality are intimately intertwined outcomes of randomly chosen measurements over entangled systems exhibiting non-local correlations are, if we preclude instantaneous influence between distant measurement choices and outcomes, random. In this paper, we survey some recent advances in the knowledge of the interplay between these two important notions from a quantum information science perspective.

  8. Community Control of Local Radio.

    ERIC Educational Resources Information Center

    Lewis, Peter M.

    This study was designed to determine to what extent local communities control the local radio which serves them, by what formal mechanisms their control is secured, and the underlying assumptions and goals which govern the practice of the professionals who have charge of the facilities. Two British radio stations, BBC Radio Bristol and the…

  9. How Just is Local Justice?

    ERIC Educational Resources Information Center

    Schultze, William A.

    1975-01-01

    Studies of criminal law practices suggest that variations in treatment from locality to locality depend on the judicial culture of the particular city, socioeconomic status of the accused, and the attitudes and actions of the police. This journal is available from the Law in American Society Foundation, 33 North LaSalle Street, Suite 1700; Chicago…

  10. Balancing State and Local Assessments.

    ERIC Educational Resources Information Center

    Rabinowitz, Stanley

    2001-01-01

    Describes rationale and attributes of local assessment program to augment statewide testing programs. Describes several steps to develop and implement a local assessment program, including setting priorities and goals, meeting with state officials, developing budgets and funding sources, forming development teams, providing professional…

  11. Developing Local Lifelong Guidance Strategies.

    ERIC Educational Resources Information Center

    Watts, A. G.; Hawthorn, Ruth; Hoffbrand, Jill; Jackson, Heather; Spurling, Andrea

    1997-01-01

    Outlines the background, rationale, methodology, and outcomes of developing local lifelong guidance strategies in four geographic areas. Analyzes the main components of the strategies developed and addresses a number of issues relating to the process of strategy development. Explores implications for parallel work in other localities. (RJM)

  12. Student Papers in Local History.

    ERIC Educational Resources Information Center

    Johnson County Community Coll., Overland Park, KS. Johnson County Center for Local History.

    Thirteen papers on Kansas and Johnson County, Kansas history are presented. The papers were written by students in a course at the Johnson County Center for Local History or for independent study in local history. The papers are: "Conditions and Construction of Gardner Lake"; "The History of St. Joseph's Church, Shawnee,…

  13. Volunteers Help Stretch Local Budgets.

    ERIC Educational Resources Information Center

    Valente, Maureen Godsey

    1985-01-01

    Discusses use of volunteers to augment local government services such as libraries, parks, paralegal aid, elderly care, data processing. Outlines requirements of successful programs and steps toward initiating volunteer programs. Presents case studies of volunteer programs in two Maryland communities and 41 examples of how local governments can…

  14. Local temperatures and local terms in modular Hamiltonians

    NASA Astrophysics Data System (ADS)

    Arias, Raúl E.; Blanco, David D.; Casini, Horacio; Huerta, Marina

    2017-03-01

    We show there are analogs to the Unruh temperature that can be defined for any quantum field theory and region of the space. These local temperatures are defined using relative entropy with localized excitations. We show that important restrictions arise from relative entropy inequalities and causal propagation between Cauchy surfaces. These suggest a large amount of universality for local temperatures, especially the ones affecting null directions. For regions with any number of intervals in two spacetime dimensions, the local temperatures might arise from a term in the modular Hamiltonian proportional to the stress tensor. We argue this term might be universal, with a coefficient that is the same for any theory, and check analytically and numerically that this is the case for free massive scalar and Dirac fields. In dimensions d ≥3 , the local terms in the modular Hamiltonian producing these local temperatures cannot be formed exclusively from the stress tensor. For a free scalar field, we classify the structure of the local terms.

  15. Localization scheme for relativistic spinors

    NASA Astrophysics Data System (ADS)

    Ciupka, J.; Hanrath, M.; Dolg, M.

    2011-12-01

    A new method to determine localized complex-valued one-electron functions in the occupied space is presented. The approach allows the calculation of localized orbitals regardless of their structure and of the entries in the spinor coefficient matrix, i.e., one-, two-, and four-component Kramers-restricted or unrestricted one-electron functions with real or complex expansion coefficients. The method is applicable to localization schemes that maximize (or minimize) a functional of the occupied spinors and that use a localization operator for which a matrix representation is available. The approach relies on the approximate joint diagonalization (AJD) of several Hermitian (symmetric) matrices which is utilized in electronic signal processing. The use of AJD in this approach has the advantage that it allows a reformulation of the localization criterion on an iterative 2 × 2 pair rotating basis in an analytical closed form which has not yet been described in the literature for multi-component (complex-valued) spinors. For the one-component case, the approach delivers the same Foster-Boys or Pipek-Mezey localized orbitals that one obtains from standard quantum chemical software, whereas in the multi-component case complex-valued spinors satisfying the selected localization criterion are obtained. These localized spinors allow the formulation of local correlation methods in a multi-component relativistic framework, which was not yet available. As an example, several heavy and super-heavy element systems are calculated using a Kramers-restricted self-consistent field and relativistic two-component pseudopotentials in order to investigate the effect of spin-orbit coupling on localization.

  16. An Open Localization and Local Communication Embodied Sensor

    PubMed Central

    Gutiérrez, Álvaro; Campo, Alexandre; Dorigo, Marco; Amor, Daniel; Magdalena, Luis; Félix, Monasterio-Huelin

    2008-01-01

    In this paper we describe a localization and local communication system which allows situated agents to communicate locally, obtaining at the same time both the range and the bearing of the emitter without the need of any centralized control or any external reference. The system relies on infrared communications with frequency modulation and is composed of two interconnected modules for data and power measurement. Thanks to the open hardware license under which it is released, the research community can easily replicate the system at a low cost and/or adapt it for applications in sensor networks and in robotics. PMID:27873944

  17. An Open Localization and Local Communication Embodied Sensor.

    PubMed

    Gutiérrez, Alvaro; Campo, Alexandre; Dorigo, Marco; Amor, Daniel; Magdalena, Luis; Monasterio-Huelin, Félix

    2008-11-25

    In this paper we describe a localization and local communication system which allows situated agents to communicate locally, obtaining at the same time both the range and the bearing of the emitter without the need of any centralized control or any external reference. The system relies on infrared communications with frequency modulation and is composed of two interconnected modules for data and power measurement. Thanks to the open hardware license under which it is released, the research community can easily replicate the system at a low cost and/or adapt it for applications in sensor networks and in robotics.

  18. Regulation of BLM Nucleolar Localization

    PubMed Central

    Tangeman, Larissa; McIlhatton, Michael A.; Grierson, Patrick; Groden, Joanna; Acharya, Samir

    2016-01-01

    Defects in coordinated ribosomal RNA (rRNA) transcription in the nucleolus cause cellular and organismal growth deficiencies. Bloom’s syndrome, an autosomal recessive human disorder caused by mutated recQ-like helicase BLM, presents with growth defects suggestive of underlying defects in rRNA transcription. Our previous studies showed that BLM facilitates rRNA transcription and interacts with RNA polymerase I and topoisomerase I (TOP1) in the nucleolus. The mechanisms regulating localization of BLM to the nucleolus are unknown. In this study, we identify the TOP1-interaction region of BLM by co-immunoprecipitation of in vitro transcribed and translated BLM segments and show that this region includes the highly conserved nuclear localization sequence (NLS) of BLM. Biochemical and nucleolar co-localization studies using site-specific mutants show that two serines within the NLS (S1342 and S1345) are critical for nucleolar localization of BLM but do not affect the functional interaction of BLM with TOP1. Mutagenesis of both serines to aspartic acid (phospho-mimetic), but not alanine (phospho-dead), results in approximately 80% reduction in nucleolar localization of BLM while retaining the biochemical functions and nuclear localization of BLM. Our studies suggest a role for this region in regulating nucleolar localization of BLM via modification of the two serines within the NLS. PMID:27657136

  19. Local Anesthetic-Induced Neurotoxicity

    PubMed Central

    Verlinde, Mark; Hollmann, Markus W.; Stevens, Markus F.; Hermanns, Henning; Werdehausen, Robert; Lirk, Philipp

    2016-01-01

    This review summarizes current knowledge concerning incidence, risk factors, and mechanisms of perioperative nerve injury, with focus on local anesthetic-induced neurotoxicity. Perioperative nerve injury is a complex phenomenon and can be caused by a number of clinical factors. Anesthetic risk factors for perioperative nerve injury include regional block technique, patient risk factors, and local anesthetic-induced neurotoxicity. Surgery can lead to nerve damage by use of tourniquets or by direct mechanical stress on nerves, such as traction, transection, compression, contusion, ischemia, and stretching. Current literature suggests that the majority of perioperative nerve injuries are unrelated to regional anesthesia. Besides the blockade of sodium channels which is responsible for the anesthetic effect, systemic local anesthetics can have a positive influence on the inflammatory response and the hemostatic system in the perioperative period. However, next to these beneficial effects, local anesthetics exhibit time and dose-dependent toxicity to a variety of tissues, including nerves. There is equivocal experimental evidence that the toxicity varies among local anesthetics. Even though the precise order of events during local anesthetic-induced neurotoxicity is not clear, possible cellular mechanisms have been identified. These include the intrinsic caspase-pathway, PI3K-pathway, and MAPK-pathways. Further research will need to determine whether these pathways are non-specifically activated by local anesthetics, or whether there is a single common precipitating factor. PMID:26959012

  20. Vibrational Spectroscopy and Quantum Localization

    NASA Astrophysics Data System (ADS)

    Fillaux, François

    These lecture-notes are meant to provide newcomers with an overview of the impact of vibrational spectroscopy in the field of nonlinear dynamics of atoms and molecules, in the perspective of energy localization. In the introduction, the terminology of nonlinear excitations and tentative experimental evidences are briefly recalled in a brief historical perspective. The basic principles of vibrational spectroscopy are presented in section 11 for infrared, Raman and inelastic neutron scattering. The potentialities for each technique to probing energy localization are discussed. In section 12, nonlinear dynamics in isolated molecules are treated within the framework of normal versus local mode representations. It is shown that these complementary representations are not necessarily distinctive of weak versus strong anharmonicity, in the context of chemical complexity. It is emphasized that local modes and energy localization are totally independent concepts. In section 4, examples of nonlinear dynamics in crystals are reviewed: multiphonon bound states, strong coupling between phonons and electrons probed with resonance Raman, local modes and quantum rotation in one-dimension probed with inelastic neutron scattering, strong coupling in hydrogen-bonded crystals and self-trapping probed with time-resolved vibrational-spectroscopy. The extended character of eigenstates in crystals free of impurities and disorder, the nature of the interaction of periodic lattices with plane waves, the Franck-Condon principle and the particle-wave duality in the quantum regime are key factors preventing observation of energy localization. It is shown that free spatially-localized nondissipative classical waves give rise to free pseudoparticles that behave as planar waves in the quantum regime. In conclusion, a clear demonstration that energy localization corresponds to eigenstates is eagerly expected for further evidencing these states with vibrational spectroscopy.

  1. Morbidity, mortality and local anaesthesia.

    PubMed

    Malamed, S F

    1999-01-01

    Two cases of local anaesthetic overdose and death are described. The patients, a four-year-old child and a 68-year old female, received local anaesthetic doses greatly in excess of those recommended. Their overdose reactions are described as well as subsequent management. The paper reviews the causes of local anaesthetic overdose, its signs and symptoms, and the recommended management of these reactions. Specific discussion as to the cause of death of these two patients and of the means to prevent such occurrences in the future concludes the paper.

  2. Speeding up local correlation methods

    SciTech Connect

    Kats, Daniel

    2014-12-28

    We present two techniques that can substantially speed up the local correlation methods. The first one allows one to avoid the expensive transformation of the electron-repulsion integrals from atomic orbitals to virtual space. The second one introduces an algorithm for the residual equations in the local perturbative treatment that, in contrast to the standard scheme, does not require holding the amplitudes or residuals in memory. It is shown that even an interpreter-based implementation of the proposed algorithm in the context of local MP2 method is faster and requires less memory than the highly optimized variants of conventional algorithms.

  3. Local governments LANDSAT applications program

    NASA Technical Reports Server (NTRS)

    1983-01-01

    The approach used to develop the internal capabilities of local governments to handle and evaluate LANDSAT data included remote sensing training, development of a low-cost digital image processing system, and technical assistance. Cost sharing, program management and coordination, and networking were also employed to address problems related to land use, water resources, environmental assessment, and air quality as experienced by urban planners. Local experiences gained in Atlanta, Georgia; Henrico County, Virginia; Oklahoma City; Oklahoma; and San Jose, California are described. Policy recommendations formulated for transferring remote sensing technologies to local governments are included.

  4. Advanced local area network concepts

    NASA Technical Reports Server (NTRS)

    Grant, Terry

    1985-01-01

    Development of a good model of the data traffic requirements for Local Area Networks (LANs) onboard the Space Station is the driving problem in this work. A parameterized workload model is under development. An analysis contract has been started specifically to capture the distributed processing requirements for the Space Station and then to develop a top level model to simulate how various processing scenarios can handle the workload and what data communication patterns result. A summary of the Local Area Network Extendsible Simulator 2 Requirements Specification and excerpts from a grant report on the topological design of fiber optic local area networks with application to Expressnet are given.

  5. Quantum localization of classical mechanics

    NASA Astrophysics Data System (ADS)

    Batalin, Igor A.; Lavrov, Peter M.

    2016-07-01

    Quantum localization of classical mechanics within the BRST-BFV and BV (or field-antifield) quantization methods are studied. It is shown that a special choice of gauge fixing functions (or BRST-BFV charge) together with the unitary limit leads to Hamiltonian localization in the path integral of the BRST-BFV formalism. In turn, we find that a special choice of gauge fixing functions being proportional to extremals of an initial non-degenerate classical action together with a very special solution of the classical master equation result in Lagrangian localization in the partition function of the BV formalism.

  6. Quantum transport localization through graphene.

    PubMed

    Srivastava, Saurabh; Kino, Hiori; Nakaharai, Shu; Verveniotis, Elisseos; Okawa, Yuji; Ogawa, Shinichi; Joachim, Christian; Aono, Masakazu

    2017-01-20

    Localization of atomic defect-induced electronic transport through a single graphene layer is calculated using a full-valence electronic structure description as a function of the defect density and taking into account the atomic-scale deformations of the layer. The elementary electronic destructive interferences leading to Anderson localization are analyzed. The low-voltage current intensity decreases with increasing length and defect density, with a calculated localization length ζ = 3.5 nm for a defect density of 5%. The difference from the experimental defect density of 0.5% required for an oxide surface-supported graphene to obtain the same ζ is discussed, pointing out how interactions of the graphene supporting surface and surface chemical modifications also control electronic transport localization.

  7. Local Authority and Town Planning.

    ERIC Educational Resources Information Center

    Duder, Bruce

    1987-01-01

    Provides an overview and definition of local authority and town planning in New Zealand. Demonstrates the relevance of planning matters to the teaching of geography. Reviews objectives of geography and specific planning methods used in several districts. (BR)

  8. Going Local to Find Help

    MedlinePlus

    ... Issues Cover Story: Traumatic Brain Injury Going Local to Find Help Past Issues / Fall 2008 Table of Contents ... description, phone numbers, maps and directions, such as To Find Out More: Visit www.ninds.nih.gov/disorders/ ...

  9. Recovering entanglement by local operations

    SciTech Connect

    D’Arrigo, A.; Lo Franco, R.; Benenti, G.; Paladino, E.; Falci, G.

    2014-11-15

    We investigate the phenomenon of bipartite entanglement revivals under purely local operations in systems subject to local and independent classical noise sources. We explain this apparent paradox in the physical ensemble description of the system state by introducing the concept of “hidden” entanglement, which indicates the amount of entanglement that cannot be exploited due to the lack of classical information on the system. For this reason this part of entanglement can be recovered without the action of non-local operations or back-transfer process. For two noninteracting qubits under a low-frequency stochastic noise, we show that entanglement can be recovered by local pulses only. We also discuss how hidden entanglement may provide new insights about entanglement revivals in non-Markovian dynamics.

  10. Local Area Networks (The Printout).

    ERIC Educational Resources Information Center

    Aron, Helen; Balajthy, Ernest

    1989-01-01

    Describes the Local Area Network (LAN), a project in which students used LAN-based word processing and electronic mail software as the center of a writing process approach. Discusses the advantages and disadvantages of networking. (MM)

  11. Composition of Local Galactic Medium

    NASA Video Gallery

    Animated view showing the neon to oxygen ratio in the neutral gas of the local cloud, as obtained with IBEX, in comparison with the ratio for the Sun and the Milky Way galaxy. There is much less ox...

  12. Nucleolar localization of myc transcripts.

    PubMed Central

    Bond, V C; Wold, B

    1993-01-01

    In situ hybridization has revealed a striking subnuclear distribution of c-myc RNA transcripts. A major fraction of the sense-strand nuclear c-myc transcripts was localized to the nucleoli. myc intron 1-containing RNAs were noticeably absent from nucleoli, accumulating instead in the nucleoplasm. The localization of myc RNA to nucleoli was shown to be common to a number of diverse cell types, including primary Sertoli cells and several cell lines. Furthermore, nucleolar localization was not restricted to c-myc and N-myc and myoD transcripts also displayed this phenomenon. In contrast, gamma-actin or lactate dehydrogenase transcripts did not display nucleolar localization. These observations suggest a new role for the nucleolus in transport and/or turnover of potential mRNAs. Images PMID:7684491

  13. Quantum transport localization through graphene

    NASA Astrophysics Data System (ADS)

    Srivastava, Saurabh; Kino, Hiori; Nakaharai, Shu; Verveniotis, Elisseos; Okawa, Yuji; Ogawa, Shinichi; Joachim, Christian; Aono, Masakazu

    2017-01-01

    Localization of atomic defect-induced electronic transport through a single graphene layer is calculated using a full-valence electronic structure description as a function of the defect density and taking into account the atomic-scale deformations of the layer. The elementary electronic destructive interferences leading to Anderson localization are analyzed. The low-voltage current intensity decreases with increasing length and defect density, with a calculated localization length ζ = 3.5 nm for a defect density of 5%. The difference from the experimental defect density of 0.5% required for an oxide surface-supported graphene to obtain the same ζ is discussed, pointing out how interactions of the graphene supporting surface and surface chemical modifications also control electronic transport localization.

  14. Fermion localization on thick branes

    SciTech Connect

    Melfo, Alejandra; Pantoja, Nelson; Tempo, Jose David

    2006-02-15

    We consider chiral fermion confinement in scalar thick branes, which are known to localize gravity, coupled through a Yukawa term. The conditions for the confinement and their behavior in the thin-wall limit are found for various different BPS branes, including double walls and branes interpolating between different AdS{sub 5} spacetimes. We show that only one massless chiral mode is localized in all these walls, whenever the wall thickness is keep finite. We also show that, independently of wall's thickness, chiral fermionic modes cannot be localized in dS{sub 4} walls embedded in a M{sub 5} spacetime. Finally, massive fermions in double wall spacetimes are also investigated. We find that, besides the massless chiral mode localization, these double walls support quasilocalized massive modes of both chiralities.

  15. Locally homogeneous pp-waves

    NASA Astrophysics Data System (ADS)

    Globke, Wolfgang; Leistner, Thomas

    2016-10-01

    We show that every n-dimensional locally homogeneous pp-wave is a plane wave, provided it is indecomposable and its curvature operator, when acting on 2-forms, has rank greater than one. As a consequence we obtain that indecomposable, Ricci-flat locally homogeneous pp-waves are plane waves. This generalises a classical result by Jordan, Ehlers and Kundt in dimension 4. Several examples show that our assumptions on indecomposability and the rank of the curvature are essential.

  16. Local-Area-Network Simulator

    NASA Technical Reports Server (NTRS)

    Gibson, Jim; Jordan, Joe; Grant, Terry

    1990-01-01

    Local Area Network Extensible Simulator (LANES) computer program provides method for simulating performance of high-speed local-area-network (LAN) technology. Developed as design and analysis software tool for networking computers on board proposed Space Station. Load, network, link, and physical layers of layered network architecture all modeled. Mathematically models according to different lower-layer protocols: Fiber Distributed Data Interface (FDDI) and Star*Bus. Written in FORTRAN 77.

  17. Local realism of macroscopic correlations.

    PubMed

    Ramanathan, R; Paterek, T; Kay, A; Kurzyński, P; Kaszlikowski, D

    2011-08-05

    We identify conditions under which correlations resulting from quantum measurements performed on macroscopic systems (systems composed of a number of particles of the order of the Avogadro number) can be described by local realism. We argue that the emergence of local realism at the macroscopic level is caused by an interplay between the monogamous nature of quantum correlations and the fact that macroscopic measurements do not reveal properties of individual particles.

  18. Local Realism of Macroscopic Correlations

    NASA Astrophysics Data System (ADS)

    Ramanathan, R.; Paterek, T.; Kay, A.; Kurzyński, P.; Kaszlikowski, D.

    2011-08-01

    We identify conditions under which correlations resulting from quantum measurements performed on macroscopic systems (systems composed of a number of particles of the order of the Avogadro number) can be described by local realism. We argue that the emergence of local realism at the macroscopic level is caused by an interplay between the monogamous nature of quantum correlations and the fact that macroscopic measurements do not reveal properties of individual particles.

  19. Monitoring cables for local degradation

    SciTech Connect

    Bustard, L.D.; Sliter, G.E.

    1989-01-01

    Recent experiences in operating nuclear plants in the United States have demonstrated the need for an in situ cable condition monitoring technique that can assess whether installed, low-voltage, unshielded cables have local damage that could compromise their ability to function under normal and accident service conditions. This paper summarizes current US programs that have been initiated to develop a technological basis for monitoring cables with local degradation. 7 refs.

  20. Cutaneous Vasodilation during Local Heating: Role of Local Cutaneous Thermosensation

    PubMed Central

    Mack, Gary W.; Foote, Kristopher M.; Nelson, W. Bradley

    2016-01-01

    We tested the hypothesis that cutaneous vasodilation during local skin heating in humans could be manipulated based upon the ability to desensitize TRPV4 ion channels by applying the thermal stimuli in a series of pulses. Each subject was instrumented with intradermal microdialysis probes in the dorsal forearm skin and perfused with 0.9% saline at 1.5 μl/min with local skin temperature controlled with a Peltier unit (9 cm2) at 34°C. Local skin temperature was manipulated for 50 min in two classic ways: a step increase to 38°C (0.1°C/s, n = 10), and a step increase to 42°C (n = 10). To desensitize TRPV4 ion channels local skin temperature was manipulated in the following way: pulsed increase to 38°C (1 pulse per min, 30 s duration, 1.0°C/s, n = 10), and 4) pulsed increase to 42°C (1.0°C/s, n = 9). Skin blood flow (SkBF, laser Doppler) was recorded directly over the middle microdialysis probe and the dialysate from all three probes were collected during baseline (34°C) and each skin heating period. The overall cutaneous vascular conductance (CVC) response to local heating was estimated from the area under the % CVCmax-time curve. The appearance of the neuropeptide calcitonin gene related peptide (CGRP) in dialysate did not change with skin heating in any protocol. For the skin temperature challenge of 34 to 38°C, the area under the % CVCmax-time curve averaged 1196 ± 295 (SD) % CVCmax•min, which was larger than the 656 ± 282% CVCmax•min during pulsed heating (p < 0.05). For the skin temperature challenge of 34 to 42°C, the area under the % CVCmax-time curve averaged 2678 ± 458% CVCmax•min, which was larger than the 1954 ± 533% CVCmax•min during pulsed heating (p < 0.05). The area under the % CVCmax•min curve, was directly proportional to the accumulated local skin thermal stress (in °C•min) (r2 = 0.62, p < 0.05, n = 39). This association indicates a critical role of local integration of thermosensitive receptors in mediating the cutaneous

  1. Calving localization at Helheim Glacier using multiple local seismic stations

    NASA Astrophysics Data System (ADS)

    Mei, M. Jeffrey; Holland, David M.; Anandakrishnan, Sridhar; Zheng, Tiantian

    2017-02-01

    A multiple-station technique for localizing glacier calving events is applied to Helheim Glacier in southeastern Greenland. The difference in seismic-wave arrival times between each pairing of four local seismometers is used to generate a locus of possible event origins in the shape of a hyperbola. The intersection of the hyperbolas provides an estimate of the calving location. This method is used as the P and S waves are not distinguishable due to the proximity of the local seismometers to the event and the emergent nature of calving signals. We find that the seismic waves that arrive at the seismometers are dominated by surface (Rayleigh) waves. The surface-wave velocity for Helheim Glacier is estimated using a grid search with 11 calving events identified at Helheim from August 2014 to August 2015. From this, a catalogue of 11 calving locations is generated, showing that calving preferentially happens at the northern end of Helheim Glacier.

  2. Local cloning of entangled states

    SciTech Connect

    Gheorghiu, Vlad; Yu Li; Cohen, Scott M.

    2010-08-15

    We investigate the conditions under which a set S of pure bipartite quantum states on a DxD system can be locally cloned deterministically by separable operations, when at least one of the states is full Schmidt rank. We allow for the possibility of cloning using a resource state that is less than maximally entangled. Our results include that: (i) all states in S must be full Schmidt rank and equally entangled under the G-concurrence measure, and (ii) the set S can be extended to a larger clonable set generated by a finite group G of order |G|=N, the number of states in the larger set. It is then shown that any local cloning apparatus is capable of cloning a number of states that divides D exactly. We provide a complete solution for two central problems in local cloning, giving necessary and sufficient conditions for (i) when a set of maximally entangled states can be locally cloned, valid for all D; and (ii) local cloning of entangled qubit states with nonvanishing entanglement. In both of these cases, we show that a maximally entangled resource is necessary and sufficient, and the states must be related to each other by local unitary 'shift' operations. These shifts are determined by the group structure, so need not be simple cyclic permutations. Assuming this shifted form and partially entangled states, then in D=3 we show that a maximally entangled resource is again necessary and sufficient, while for higher-dimensional systems, we find that the resource state must be strictly more entangled than the states in S. All of our necessary conditions for separable operations are also necessary conditions for local operations and classical communication (LOCC), since the latter is a proper subset of the former. In fact, all our results hold for LOCC, as our sufficient conditions are demonstrated for LOCC, directly.

  3. Localization and Quantitative Co-localization of Enamelin with Amelogenin

    PubMed Central

    Gallon, Victoria; Chen, Lisha; Yang, Xiudong; Moradian-Oldak, Janet

    2013-01-01

    Enamelin and amelogenin are vital proteins in enamel formation. The cooperative function of these two proteins controls crystal nucleation and morphology in vitro. We quantitatively analyzed the co-localization between enamelin and amelogenin by confocal microscopy and using two antibodies, one raised against a sequence in the porcine 32 kDa enamelin region and the other raised against full-length recombinant mouse amelogenin. We further investigated the interaction of the porcine 32 kDa enamelin and recombinant amelogenin using immuno-gold labeling. This study reports the quantitative co-localization results for postnatal days 1, 2, 3, 4, 5, 6, 7 and 8 mandibular mouse molars. We show that amelogenin and enamelin are secreted into the extracellular matrix on the cuspal slopes of the molars at day 1 and that secretion continues to at least day 8. Quantitative co-localization analysis (QCA) was performed in several different configurations using large (45 μm height, 33 μm width) and small (7 μm diameter) regions of interest to elucidate any patterns. Co-localization patterns in day 8 samples revealed that enamelin and amelogenin co-localize near the secretory face of the ameloblasts and appear to be secreted approximately in a 1:1 ratio. The degree of co-localization decreases as the enamel matures, both along the secretory face of ameloblasts and throughout the entire thickness of the enamel. Immuno-reactivity against enamelin is concentrated along the secretory face of ameloblasts, supporting the theory that this protein together with amelogenin is intimately involved in mineral induction at the beginning of enamel formation. PMID:23563189

  4. Locally adaptive document skew detection

    NASA Astrophysics Data System (ADS)

    Sauvola, Jaakko J.; Doermann, David S.; Pietikaeinen, Matti

    1997-04-01

    This paper proposes a new approach to the detection of local orientation and skew in document images. It is based on the observation that there are many documents where a single global estimate of the page skew is not sufficient. These documents require local adaptation to deal robustly with todays complex configurations of components on the page. The approach attempts to identify regions in the image which exhibit locally consistent physical properties and consistent physical properties and consistent orientation. To do this, we rapidly compute a coarse segmentation and delineate regions which differ with respect to layout and/or physical content. Each region is classified as text, graphics, mixed text/graphics, image or background using local features and additional features are extracted to estimate orientation. The local orientation decisions are propagated where appropriate to resolve ambiguity and to produce a global estimate of the skew for the page. The implementation of our algorithms is demonstrated on a set of images which have multiple regions with different orientations.

  5. Afghanistan’s Local War: Building Local Defense Forces

    DTIC Science & Technology

    2010-01-01

    Defense Forces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 CHAPTER SIX Mitigating Risks ...exploited. There are, of course, risks with any strategy, as this assessment documents. But the potential risks are outweighed by the potential...gains, especially since Afghan and NATO forces can monitor and provide oversight to local defense forces and mitigate these risks . xiii Acknowledgments

  6. Locally-Referenced Ultrasonic – LPS for Localization and Navigation

    PubMed Central

    Gualda, David; Ureña, Jesús; García, Juan C.; Lindo, Alejandro

    2014-01-01

    This paper presents a flexible deployment of ultrasonic position sensors and a novel positioning algorithm suitable for the navigation of mobile robots (MRs) in extensive indoor environments. Our proposal uses several independently-referenced local positioning systems (LPS), which means that each one of them operates within its own local reference system. In a typical layout, an indoor extensive area can be covered using just a reduced set of globally-referenced LPS (GRLPS), whose beacon positions are known to the global reference system, while the rest of the space can be covered using locally-referenced LPSs (LRLPS) that can be distributed arbitrarily. The number of LRLPS and their position can be also changed at any moment. The algorithm is composed of several Bayesian filters running in parallel, so that when an MR is under the GRLPS coverage area, its position is updated by a global filter, whereas when the MR is inside the LRLPS area, its position is updated using position increments within a local filter. The navigation algorithm has been tested by simulation and with actual data obtained using a real set of ultrasonic LPSs. PMID:25412215

  7. Localization via Automorphisms of the CARs: Local Gauge Invariance

    NASA Astrophysics Data System (ADS)

    Grundling, Hendrik; Neeb, Karl-Hermann

    2010-08-01

    The classical matter fields are sections of a vector bundle E with base manifold M, and the space L 2( E) of square integrable matter fields w.r.t. a locally Lebesgue measure on M, has an important module action of {C_b^infty(M)} on it. This module action defines restriction maps and encodes the local structure of the classical fields. For the quantum context, we show that this module action defines an automorphism group on the algebra of the canonical anticommutation relations, CAR( L 2( E)), with which we can perform the analogous localization. That is, the net structure of the CAR( L 2( E)) w.r.t. appropriate subsets of M can be obtained simply from the invariance algebras of appropriate subgroups. We also identify the quantum analogues of restriction maps, and as a corollary, we prove a well-known “folk theorem,” that the CAR( L 2( E)) contains only trivial gauge invariant observables w.r.t. a local gauge group acting on E.

  8. Sound Localization in the Alligator

    PubMed Central

    Carr, Catherine E.

    2016-01-01

    In early tetrapods, it is assumed that the tympana were acoustically coupled through the pharynx and therefore inherently directional, acting as pressure difference receivers. The later closure of the middle ear cavity in turtles, archosaurs, and mammals is a derived condition, and would have changed the ear by decoupling the tympana. Isolation of the middle ears would then have led to selection for structural and neural strategies to compute sound source localization in both archosaurs and mammalian ancestors. In the archosaurs (birds and crocodilians) the presence of air spaces in the skull provided connections between the ears that have been exploited to improve directional hearing, while neural circuits mediating sound localization are well developed. In this review, we will focus primarily on directional hearing in crocodilians, where vocalization and sound localization are thought to be ecologically important, and indicate important issues still awaiting resolution. PMID:26048335

  9. Dermatomycosis Frequency and Localization Sites

    PubMed Central

    Koçinaj, Allma Ferizi; Kotori, Merita Grajqevci; Koraqi, Andi; Fida, Monika

    2015-01-01

    Introduction: Since the prevalence of skin mycotic infections is changing and is area depended we aimed to analyze the frequency of the skin myocotic infections and the appearance sites. Material and Methods: There were involved 560 patients referred to the Dermatology Clinic of University Clinical Center of Kosova during a period of one year. Results: The mean age of our study group was around thirties with a predominance of female and rural patients. Although most of cases presented with single site disease localization, we observed the increase in number of cases with more than one site localization with age. Conclusion: The increased prevalence skin mycotic infections, as well as more than one place of localization deserve a multidimensional approach. PMID:25870481

  10. Boundary Preserving Dense Local Regions.

    PubMed

    Kim, Jaechul; Grauman, Kristen

    2015-05-01

    We propose a dense local region detector to extract features suitable for image matching and object recognition tasks. Whereas traditional local interest operators rely on repeatable structures that often cross object boundaries (e.g., corners, scale-space blobs), our sampling strategy is driven by segmentation, and thus preserves object boundaries and shape. At the same time, whereas existing region-based representations are sensitive to segmentation parameters and object deformations, our novel approach to robustly sample dense sites and determine their connectivity offers better repeatability. In extensive experiments, we find that the proposed region detector provides significantly better repeatability and localization accuracy for object matching compared to an array of existing feature detectors. In addition, we show our regions lead to excellent results on two benchmark tasks that require good feature matching: weakly supervised foreground discovery and nearest neighbor-based object recognition.

  11. Localized structures in convective experiments

    NASA Astrophysics Data System (ADS)

    Burguete, J.; Mancini, H.

    2014-01-01

    In this work we review localized structures appearing in thermo-convective experiments performed in extended (large "aspect ratio") fluid layers. After a brief general review (not exhaustive), we focus on some results obtained in pure fluids in a Bénard-Marangoni system with non-homogeneous heating where some structures of this kind appear. The experimental results are compared in reference to the most classical observed in binary mixtures experiments or simulations. In the Bénard-Marangoni experiment we present the stability diagram where localized structures appear and the typical situations where these local mechanisms have been studied experimentally. Some new experimental results are also included. The authors want to honor Prof. H. Brand in his 60th. birthday and to thank him for helpful discussions.

  12. LULUs: locally unwanted land uses

    SciTech Connect

    Popper, F.J.

    1983-06-01

    A LULU is a locally unwanted land use. It may be an old-age home or a nuclear-waste-disposal site. People need it but do not want to live next to it. Some characteristics LULUs have in common are: opposition (more or less organized), costs to the neighborhood (real or perceived), support from conservatives for LULUs of the right, support from liberals for LULUs of the left, and some local support. Today's LULU may be tomorrow's prize; witness the 1982 competition for a state prison by 21 towns in depressed Illinois. Regional and national LULUs, while offering (or appearing to offer) a regional or national benefit, put financial and environmental costs and social stresses on a locality. Governmental and legal questions confront the decision-makers who must untangle these conflicts.

  13. Sound localization in the alligator.

    PubMed

    Bierman, Hilary S; Carr, Catherine E

    2015-11-01

    In early tetrapods, it is assumed that the tympana were acoustically coupled through the pharynx and therefore inherently directional, acting as pressure difference receivers. The later closure of the middle ear cavity in turtles, archosaurs, and mammals is a derived condition, and would have changed the ear by decoupling the tympana. Isolation of the middle ears would then have led to selection for structural and neural strategies to compute sound source localization in both archosaurs and mammalian ancestors. In the archosaurs (birds and crocodilians) the presence of air spaces in the skull provided connections between the ears that have been exploited to improve directional hearing, while neural circuits mediating sound localization are well developed. In this review, we will focus primarily on directional hearing in crocodilians, where vocalization and sound localization are thought to be ecologically important, and indicate important issues still awaiting resolution.

  14. Local non-equilibrium thermodynamics

    PubMed Central

    Jinwoo, Lee; Tanaka, Hajime

    2015-01-01

    Local Shannon entropy lies at the heart of modern thermodynamics, with much discussion of trajectory-dependent entropy production. When taken at both boundaries of a process in phase space, it reproduces the second law of thermodynamics over a finite time interval for small scale systems. However, given that entropy is an ensemble property, it has never been clear how one can assign such a quantity locally. Given such a fundamental omission in our knowledge, we construct a new ensemble composed of trajectories reaching an individual microstate, and show that locally defined entropy, information, and free energy are properties of the ensemble, or trajectory-independent true thermodynamic potentials. We find that the Boltzmann-Gibbs distribution and Landauer's principle can be generalized naturally as properties of the ensemble, and that trajectory-free state functions of the ensemble govern the exact mechanism of non-equilibrium relaxation. PMID:25592077

  15. Local Flaps of The Hand

    PubMed Central

    Rehim, Shady A.; Chung, Kevin C.

    2014-01-01

    Synopsis A local flap consists of skin and subcutaneous tissue that is harvested from a site nearby a given defect while maintaining its intrinsic blood supply. When a soft tissue defect of the hand is not amenable to primary closure or skin grafting, local skin flaps can be a used as a reliable source of soft tissue replacement that replaces like with like. Flaps are categorized based on their composition, method of transfer, flap design and blood supply, yet flap circulation is considered the most critical factor for the flap survival. This article reviews the classification of local skin flaps of the hand and offers a practical reconstructive approach for several soft tissue defects of the hand and digits. PMID:24731606

  16. Foreign English Language Teachers' Local Pedagogy

    ERIC Educational Resources Information Center

    Eusafzai, Hamid Ali Khan

    2015-01-01

    ELT methods have been criticized for being limited and inadequate. Postmethod pedagogy has been offered as an alternate to these methods. The postmethod pedagogy emphasises localization of pedagogy and celebrates local culture, teachers and knowledge. Localizing pedagogy is easy for local teachers as knowledge and understanding of the local comes…

  17. Spanning the Local Government Information Gap.

    ERIC Educational Resources Information Center

    Durrance, Joan C.

    1985-01-01

    Discussion of implications of highly decentralized nature of local government for collection of local documents highlights meaning of access, effect of local government environment on access to local government information, library responses, and tools that can assist libraries in increasing citizen and government access to local government…

  18. [Emergencies evolving from local anesthesia].

    PubMed

    Kaufman, E; Garfunkel, A; Findler, M; Elad, S; Zusman, S P; Malamed, S F; Galili, D

    2002-01-01

    Local anesthesia is without doubt the most frequently used drug in dentistry and in medicine. In spite of records of safety set by using these drugs, there is evidence to adverse reactions ranging from 2.5%-11%. Most of the reactions originate from the autonomic system. A recent, well-planned study indicates that adverse reactions are highly correlated to the medical status of the patient: the higher the medical risk, the greater the chance to experience an adverse reaction. This study also found that adverse reactions highly correlated to the concentration of adrenalin. Another recent study found a direct relationship between adverse reactions and the level of anxiety experienced by the patient and to the dental procedure. Most of the reactions in this study occurred either immediately at injection time and within 2 hours following the injection. Since the beginning of last century, vasoconstrictors have been added to local anesthesia solutions in order to reduce toxicity and prologue activity of the LA. However, today it is commonly agreed that this addition to local anesthesia should not be administered to cardiac patients especially those suffering from refractory dysrhythmias, angina pectoris, post myocardial infarction (6 months) and uncontrolled hypertension. Other contraindications to vasoconstrictors are endocrine disorders such as hyperthyroidism, hyperfunction of the medullary adrenal (pheochromocytoma) and uncontrolled diabetes mellitus. Cross reactivity of local anesthetic solutions can occur with MAO inhibitors, non specific beta adrenergic blockers, tricyclic antidepressants, phenothiazides and cocaine abusers. Noradrenaline added to local anesthetics as a vasoconstrictor has been described as a trigger to a great increase in blood pressure and therefore has been forbidden for use in many countries. This paper describes 4 cases of severe complications following the injections of local anesthesia of which three ended in fatality.

  19. Local management of rectal neoplasia.

    PubMed

    Touzios, John; Ludwig, Kirk A

    2008-11-01

    The treatment of rectal neoplasia, whether benign or malignant, challenges the surgeon. The challenge in treating rectal cancer is selecting the proper approach for the appropriate patient. In a small number of rectal cancer patients local excision may be the best approach. In an attempt to achieve two goals-cure of disease with a low rate of local failure and maintenance of function and quality of life-multiple approaches can be utilized. The key to obtaining a good outcome for any one patient is balancing the competing factors that impact on these goals. Any effective treatment aimed at controlling rectal cancer in the pelvis must take into account the disease in the bowel wall itself and the disease, or potential disease, in the mesorectum. The major downside of local excision techniques is the potential of leaving untreated disease in the mesorectum. Local management techniques avoid the potential morbidity, mortality, and functional consequences of a major abdominal radical resection and are thus quite effective in achieving the maintenance of function and quality of life goal. The issue for the transanal techniques is how they fare in achieving the first goal-cure of the cancer while keeping local recurrence rates to an absolute minimum. Without removing both the rectum and the mesorectum there is no completely accurate way to determine whether a rectal cancer has moved outside the bowel wall, so any decision on local management of a rectal neoplasm is a calculated risk. For benign neoplasia, the challenge is removing the lesion without having to resort to a major abdominal procedure.

  20. Biodiversity conservation in local planning.

    PubMed

    Miller, James R; Groom, Martha; Hess, George R; Steelman, Toddi; Stokes, David L; Thompson, Jan; Bowman, Troy; Fricke, Laura; King, Brandon; Marquardt, Ryan

    2009-02-01

    Local land-use policy is increasingly being recognized as fundamental to biodiversity conservation in the United States. Many planners and conservation scientists have called for broader use of planning and regulatory tools to support the conservation of biodiversity at local scales. Yet little is known about the pervasiveness of these practices. We conducted an on-line survey of county, municipal, and tribal planning directors (n =116) in 3 geographic regions of the United States: metropolitan Seattle, Washington; metropolitan Des Moines, Iowa; and the Research Triangle, North Carolina. Our objectives were to gauge the extent to which local planning departments address biodiversity conservation and to identify factors that facilitate or hinder conservation actions in local planning. We found that biodiversity conservation was seldom a major consideration in these departments. Staff time was mainly devoted to development mandates and little time was spent on biodiversity conservation. Regulations requiring conservation actions that might benefit biodiversity were uncommon, with the exception of rules governing water quality in all 3 regions and the protection of threatened and endangered species in the Seattle region. Planning tools that could enhance habitat conservation were used infrequently. Collaboration across jurisdictions was widespread, but rarely focused on conservation. Departments with a conservation specialist on staff tended to be associated with higher levels of conservation actions. Jurisdictions in the Seattle region also reported higher levels of conservation action, largely driven by state and federal mandates. Increased funding was most frequently cited as a factor that would facilitate greater consideration of biodiversity in local planning. There are numerous opportunities for conservation biologists to play a role in improving conservation planning at local scales.

  1. Operator Localization of Virtual Objects

    NASA Technical Reports Server (NTRS)

    Ellis, Stephen R.; Menges, Brian M.; Null, Cynthia H. (Technical Monitor)

    1998-01-01

    Errors in the localization of nearby virtual objects presented via see-through, helmet mounted displays are examined as a function of viewing conditions and scene content. Monocular, biocular or stereoscopic presentation of the virtual objects, accommodation (required focus), subjects'age, and the position of physical surfaces are examined. Nearby physical surfaces are found to introduce localization errors that differ depending upon the other experimental factors. The apparent physical size and transparency of the virtual objects and physical surfaces respectively are also influenced by their relative position when superimposed. Design implications are discussed.

  2. Wedge locality and asymptotic commutativity

    NASA Astrophysics Data System (ADS)

    Soloviev, M. A.

    2014-05-01

    In this paper, we study twist deformed quantum field theories obtained by combining the Wightman axiomatic approach with the idea of spacetime noncommutativity. We prove that the deformed fields with deformation parameters of opposite sign satisfy the condition of mutual asymptotic commutativity, which was used earlier in nonlocal quantum field theory as a substitute for relative locality. We also present an improved proof of the wedge localization property discovered for the deformed fields by Grosse and Lechner, and we show that the deformation leaves the asymptotic behavior of the vacuum expectation values in spacelike directions substantially unchanged.

  3. Observations of Local Seychelles Circulation

    DTIC Science & Technology

    2015-09-30

    Identification of physical oceanographic controls on mass and momentum transport on scales that are relevant to local ecology and fisheries and that...of basic regional modeling capacity that Seychelles managers and fisheries can use to guide decisions and improve community outreach and education

  4. Local Initiative for Economic Development.

    ERIC Educational Resources Information Center

    Zeiss, P. Anthony

    In fall 1982, in an effort to address a rising unemployment rate in its service area, Pueblo Community College (PCC) joined with local community leaders and the Pueblo Economic Development Corporation to explore feasible, practical, and workable assistance models which would foster the retention and development of small businesses. Numerous…

  5. Localized coherence of freak waves

    NASA Astrophysics Data System (ADS)

    Latifah, Arnida L.; van Groesen, E.

    2016-09-01

    This paper investigates in detail a possible mechanism of energy convergence leading to freak waves. We give examples of a freak wave as a (weak) pseudo-maximal wave to illustrate the importance of phase coherence. Given a time signal at a certain position, we identify parts of the time signal with successive high amplitudes, so-called group events, that may lead to a freak wave using wavelet transform analysis. The local coherence of the critical group event is measured by its time spreading of the most energetic waves. Four types of signals have been investigated: dispersive focusing, normal sea condition, thunderstorm condition and an experimental irregular wave. In all cases presented in this paper, it is shown that a high correlation exists between the local coherence and the appearance of a freak wave. This makes it plausible that freak waves can be developed by local interactions of waves in a wave group and that the effect of waves that are not in the immediate vicinity is minimal. This indicates that a local coherence mechanism within a wave group can be one mechanism that leads to the appearance of a freak wave.

  6. Superbubbles and Local Cosmic Rays

    NASA Technical Reports Server (NTRS)

    Streitmatter, Robert E.; Jones, Frank C.

    2005-01-01

    We consider the possibility that distinctive features of the local cosmic ray spectra and composition are influenced by the Solar system being embedded within the cavity of an ancient superbubble. Shifts in the measured cosmic ray composition between 10(exp 11) and 10(exp 20) eV as well as the "knee" and "second knee" may be understood in this picture.

  7. Anisotropic weak localization of light.

    PubMed

    Sapienza, Riccardo; Mujumdar, Sushil; Cheung, Cecil; Yodh, A G; Wiersma, Diederik

    2004-01-23

    We have observed angular anisotropy in weak localization of light from highly scattering, orientationally ordered, nematic liquid crystals. This demonstration of angular anisotropy in a multiple-scattering interference phenomenon was facilitated by a light scattering instrument with extraordinary angular resolution. The measured anisotropies were consistent with a simple model of coherent backscattering generalized for propagation-direction dependent mean free paths.

  8. Research for Locally Relevant Solutions

    ERIC Educational Resources Information Center

    Association of Universities and Colleges of Canada, 2004

    2004-01-01

    In the CIDA-funded University Partnerships in Cooperation and Development program--where Canadian universities establish knowledge partnerships with Southern universities--projects with a well-developed research dimension have proven to be the strongest projects, with broader and deeper contributions to the local institutions and larger community.…

  9. Universities: Engaging with Local Communities

    ERIC Educational Resources Information Center

    Universities UK, 2010

    2010-01-01

    This leaflet illustrates the many ways in which universities impact on the local area. Universities are a major contributor to the economy in their own right, both as employers and purchasers of goods. Their social and cultural influence is also felt through their provision of: (1) art galleries, museums and exhibitions; (2) cinemas and theatres;…

  10. Local Control of Blood Flow

    ERIC Educational Resources Information Center

    Clifford, Philip S.

    2011-01-01

    Organ blood flow is determined by perfusion pressure and vasomotor tone in the resistance vessels of the organ. Local factors that regulate vasomotor tone include myogenic and metabolic autoregulation, flow-mediated and conducted responses, and vasoactive substances released from red blood cells. The relative importance of each of these factors…

  11. Local Area Networks: Part II.

    ERIC Educational Resources Information Center

    Dessy, Raymond E., Ed.

    1982-01-01

    Discusses five approaches used by industry/colleges to provide local area network (LAN) capabilities in the analytical laboratory: (1) mixed baseband bus network coupled to a star net; (2) broadband bus network; (3) ring network; (4) star network coupled to broadband net; and (5) simple multiprocessor center. Part I (September issue) focused on…

  12. Local Area Networks: Part I.

    ERIC Educational Resources Information Center

    Dessy, Raymond E.

    1982-01-01

    Local area networks are common communication conduits allowing various terminals, computers, discs, printers, and other electronic devices to intercommunicate over short distances. Discusses the vocabulary of such networks including RS-232C point-to-point and IEEE-488 multidrop protocols; error detection; message packets; multiplexing; star, ring,…

  13. Weak localization of seismic waves.

    PubMed

    Larose, E; Margerin, L; Van Tiggelen, B A; Campillo, M

    2004-07-23

    We report the observation of weak localization of seismic waves in a natural environment. It emerges as a doubling of the seismic energy around the source within a spot of the width of a wavelength, which is several tens of meters in our case. The characteristic time for its onset is the scattering mean-free time that quantifies the internal heterogeneity.

  14. Collaboration with the Local Community.

    ERIC Educational Resources Information Center

    Jackson, Michael L.; Cherrey, Cynthia

    2002-01-01

    Colleges and universities continually search for ways to enhance the safety and security of their educational programs and physical plant. This article examines how the University of Southern California and other institutions are using collaborative efforts with the local community to enhance their mutual safety and security through dynamic…

  15. Local Evaluation of Chemistry Journals

    ERIC Educational Resources Information Center

    Kraus, Joseph R.; Hansen, Rachel

    2008-01-01

    This paper reports on the evaluation of local usage statistics of a specific set of chemistry journals at the University of Denver in Colorado, USA. The objective of the study is to demonstrate that commercial publishers in chemistry charge considerably more for their journals than those from the non-commercial sector. There are three variables…

  16. Local Decisions and Global Networks

    ERIC Educational Resources Information Center

    King, David C.; Long, Cathryn J.

    1976-01-01

    Impact of economic and urban planning on the natural environment can be studied through local situations: California conservation students realized the detrimental effects of a seemingly beneficial dam project. Students were inspired to initiate community-state action to correct damage to wildlife, sanitation, and farming. (AV)

  17. Local shape of pictorial relief

    PubMed Central

    Koenderink, Jan; van Doorn, Andrea; Wagemans, Johan

    2014-01-01

    How is pictorial relief represented in visual awareness? Certainly not as a “depth map,” but perhaps as a map of local surface attitudes (Koenderink & van Doorn, 1995). Here we consider the possibility that observers might instead, or concurrently, represent local surface shape, a geometrical invariant with respect to motions. Observers judge local surface shape, in a picture of a piece of sculpture, on a five-point categorical scale. Categories are cap–ridge–saddle–rut–cup–flat, where “flat” denotes the absence of shape. We find that observers readily perform such a task, with full resolution of a shape index scale (cap–ridge–saddle–rut–cup), and with excellent self-consistency over days. There exist remarkable inter-observer differences. Over a group of 10 naive observers we find that the dispersion of judgments peaks at the saddle category. There may be a relation of this finding to the history of the topic—Alberti's (1827) omission of the saddle category in his purportedly exhaustive catalog of local surface shapes. PMID:25469225

  18. Arthur Smith, Local Baptist Pastor.

    ERIC Educational Resources Information Center

    Heaton, Moss, Ed.

    1985-01-01

    Written and published by the students at Gary High School this volume has three articles dealing with East Texas life. The first "Arthur Smith" (David Hancock and others) is an account of growing up in Marian County, Texas is described by the local Baptist minister. The pastor begins with the year of his birth and gives detailed…

  19. Localization model for cochlear implants

    NASA Astrophysics Data System (ADS)

    Miller, Douglas A.; Matin, Mohammad A.

    2011-09-01

    Normal hearing persons are able to localize the direction of sounds better using both ears than when listening with only one ear. Localization ability is dependent on auditory system perception of interaural differences in time, intensity, and phase. Interaural timing differences (ITDs) provide information for locating direction of low and mid frequency sounds, while interaural level differences (ILDs), which occur because of the horizontal plane shadowing effect of the head, provide information for locating direction of higher frequency sounds. The head related transfer function (HRTF) contains characteristic information important for acoustic localization. Models based on HRTFs take into account head shadow, torso, and pinna effects, and their impact on interaural frequency, level, and timing differences. Cochlear implants (CIs) have proven a successful treatment for persons with bilateral severe to profound hearing loss. A problem is that only some ITD and ILD cues are maintained with CI sound processing, and the microphone position alters the acoustic cues. The relative impact of differences in physical cues received by the auditory system with bilateral CIs versus differences in the ability of the damaged auditory nervous system to process bilateral inputs is not yet clear. The model presented in this paper was constructed as a step toward answering this question, and is intended to serve as a tool for future development of more optimal signal processing algorithms that may provide better localization ability for persons with bilateral CIs.

  20. Regularization of Localized Degradation Processes

    DTIC Science & Technology

    1996-12-28

    order to assess the regularization properties of non-classical micropolar Cosserat continua which feature non-symmetric stress and strain tensors because...of the presence of couple-stresses and micro-curvatures. It was shown that micropolar media may only exhibit localized failure in the form of tensile

  1. Localized gravity in string theory.

    PubMed

    Karch, A; Randall, L

    2001-08-06

    We propose a string realization of the AdS4 brane in AdS5 that is known to localize gravity. Our theory is M D5 branes in the near horizon geometry of N D3 branes, where M and N are appropriately tuned.

  2. Sound localization by echolocating bats

    NASA Astrophysics Data System (ADS)

    Aytekin, Murat

    Echolocating bats emit ultrasonic vocalizations and listen to echoes reflected back from objects in the path of the sound beam to build a spatial representation of their surroundings. Important to understanding the representation of space through echolocation are detailed studies of the cues used for localization, the sonar emission patterns and how this information is assembled. This thesis includes three studies, one on the directional properties of the sonar receiver, one on the directional properties of the sonar transmitter, and a model that demonstrates the role of action in building a representation of auditory space. The general importance of this work to a broader understanding of spatial localization is discussed. Investigations of the directional properties of the sonar receiver reveal that interaural level difference and monaural spectral notch cues are both dependent on sound source azimuth and elevation. This redundancy allows flexibility that an echolocating bat may need when coping with complex computational demands for sound localization. Using a novel method to measure bat sonar emission patterns from freely behaving bats, I show that the sonar beam shape varies between vocalizations. Consequently, the auditory system of a bat may need to adapt its computations to accurately localize objects using changing acoustic inputs. Extra-auditory signals that carry information about pinna position and beam shape are required for auditory localization of sound sources. The auditory system must learn associations between extra-auditory signals and acoustic spatial cues. Furthermore, the auditory system must adapt to changes in acoustic input that occur with changes in pinna position and vocalization parameters. These demands on the nervous system suggest that sound localization is achieved through the interaction of behavioral control and acoustic inputs. A sensorimotor model demonstrates how an organism can learn space through auditory-motor contingencies

  3. Local flicker stimulation evokes local retinal blood velocity changes

    PubMed Central

    Zhong, Zhangyi; Huang, Gang; Chui, Toco Yuen Ping; Petrig, Benno L.; Burns, Stephen A.

    2012-01-01

    We investigated the effect of localized visual stimulation on human retinal blood velocity using an adaptive optics scanning laser ophthalmoscope (AOSLO). To measure the blood velocity response, the AOSLO scanning raster was moved over the target arteries and red blood cell velocity was measured. Localized visual stimuli were delivered by projecting flicker patterns inside or outside the target artery's downstream region. The blood velocity increased in the presence of a flicker stimulus in the downstream region but not when outside the downstream region. The blood velocity increased more with larger area of stimulation. This increase was significant even when the stimulus was smaller than 600 μm × 600 μm. These findings suggest that when the retina regulates its blood flow to metabolic demands, it regulates blood velocity in the vascular system selectively, according to activity of neurons within its field of influence. PMID:22661609

  4. Comments on localized and integral localized approximations in spherical coordinates

    NASA Astrophysics Data System (ADS)

    Gouesbet, Gérard; Lock, James A.

    2016-08-01

    Localized approximation procedures are efficient ways to evaluate beam shape coefficients of laser beams, and are particularly useful when other methods are ineffective or inefficient. Comments on these procedures are, however, required in order to help researchers make correct decisions concerning their use. This paper has the flavor of a short review and takes the opportunity to attract the attention of the readers to a required refinement of terminology.

  5. Sacroplasty for Local or Massive Localization of Multiple Myeloma

    SciTech Connect

    Basile, Antonio; Tsetis, Dimitrios; Cavalli, Maide; Fiumara, Paolo; Raimondo, Francesco Di; Coppolino, Francesco; Coppolino, Carmelo; Mundo, Elena; Desiderio, Carla; Granata, Antonio; Patti, Maria Teresa

    2010-12-15

    The purpose of this study was to assess the efficacy of cementoplasty in the treatment of sacral multiple myelomas. We retrospectively reviewed the records of eight patients (four women and four men; age range 47-68 years; mean age 57.8) who underwent cementoplasty for painful osteolytic localization of multiple myeloma between April 2007 and May 2009. The patients had difficulty walking because of increasing pain. Six patients had persistent pain despite other cementoplasties for vertebral and femoral localization, whereas two patients referred at the time of diagnosis had only sacral lesions. The clinical indication for treatment was (1) a pain intensity score {>=}5 on visual analogue scale (VAS) and (2) pain totally or partially refractory to analgesic treatment in patients with a life expectancy >3 months. Technical planning was based on computed tomography and/or magnetic resonance imaging. Six patients had previously undergone radiotherapy or chemotherapy and were receiving varying doses of analgesics, whereas sacroplasty represented the first treatment for two patients. Five patients had monolateral local involvement, and the other patients had massive involvement of the sacrum; Technical success was achieved in all cases. We had only one small and asymptomatic foraminal leak. All patients experienced improvement in symptoms after the procedure, as demonstrated by improved VAS scores and performance status (PS) and decreased analgesic dose constant during follow-up. In our experience, percutaneous stabilization can be used effectively and safely in patients with focal or extensive involvement of the sacrum by multiple myeloma.

  6. Dynamically screened local correlation method using enveloping localized orbitals.

    PubMed

    Auer, Alexander A; Nooijen, Marcel

    2006-07-14

    In this paper we present a local coupled cluster approach based on a dynamical screening scheme, in which amplitudes are either calculated at the coupled cluster level (in this case CCSD) or at the level of perturbation theory, employing a threshold driven procedure based on MP2 energy increments. This way, controllable accuracy and smooth convergence towards the exact result are obtained in the framework of an a posteriori approximation scheme. For the representation of the occupied space a new set of local orbitals is presented with the size of a minimal basis set. This set is atom centered, is nonorthogonal, and has shapes which are fairly independent of the details of the molecular system of interest. Two slightly different versions of combined local coupled cluster and perturbation theory equations are considered. In the limit both converge to the untruncated CCSD result. Benchmark calculations for four systems (heptane, serine, water hexamer, and oxadiazole-2-oxide) are carried out, and decay of the amplitudes, truncation error, and convergence towards the exact CCSD result are analyzed.

  7. Dynamically screened local correlation method using enveloping localized orbitals

    NASA Astrophysics Data System (ADS)

    Auer, Alexander A.; Nooijen, Marcel

    2006-07-01

    In this paper we present a local coupled cluster approach based on a dynamical screening scheme, in which amplitudes are either calculated at the coupled cluster level (in this case CCSD) or at the level of perturbation theory, employing a threshold driven procedure based on MP2 energy increments. This way, controllable accuracy and smooth convergence towards the exact result are obtained in the framework of an a posteriori approximation scheme. For the representation of the occupied space a new set of local orbitals is presented with the size of a minimal basis set. This set is atom centered, is nonorthogonal, and has shapes which are fairly independent of the details of the molecular system of interest. Two slightly different versions of combined local coupled cluster and perturbation theory equations are considered. In the limit both converge to the untruncated CCSD result. Benchmark calculations for four systems (heptane, serine, water hexamer, and oxadiazole-2-oxide) are carried out, and decay of the amplitudes, truncation error, and convergence towards the exact CCSD result are analyzed.

  8. Fractal superconductivity near localization threshold

    SciTech Connect

    Feigel'man, M.V.; Ioffe, L.B.; Kravtsov, V.E.; Cuevas, E.

    2010-07-15

    We develop a semi-quantitative theory of electron pairing and resulting superconductivity in bulk 'poor conductors' in which Fermi energy E{sub F} is located in the region of localized states not so far from the Anderson mobility edge E{sub c}. We assume attractive interaction between electrons near the Fermi surface. We review the existing theories and experimental data and argue that a large class of disordered films is described by this model. Our theoretical analysis is based on analytical treatment of pairing correlations, described in the basis of the exact single-particle eigenstates of the 3D Anderson model, which we combine with numerical data on eigenfunction correlations. Fractal nature of critical wavefunction's correlations is shown to be crucial for the physics of these systems. We identify three distinct phases: 'critical' superconductive state formed at E{sub F} = E{sub c}, superconducting state with a strong pseudo-gap, realized due to pairing of weakly localized electrons and insulating state realized at E{sub F} still deeper inside a localized band. The 'critical' superconducting phase is characterized by the enhancement of the transition temperature with respect to BCS result, by the inhomogeneous spatial distribution of superconductive order parameter and local density of states. The major new feature of the pseudo-gapped state is the presence of two independent energy scales: superconducting gap {Delta}, that is due to many-body correlations and a new 'pseudo-gap' energy scale {Delta}{sub P} which characterizes typical binding energy of localized electron pairs and leads to the insulating behavior of the resistivity as a function of temperature above superconductive T{sub c}. Two gap nature of the pseudo-gapped superconductor is shown to lead to specific features seen in scanning tunneling spectroscopy and point-contact Andreev spectroscopy. We predict that pseudo-gapped superconducting state demonstrates anomalous behavior of the optical

  9. Local resection of ampullary tumors.

    PubMed

    Meneghetti, Adam T; Safadi, Bassem; Stewart, Lygia; Way, Lawrence W

    2005-12-01

    There is no consensus on the appropriateness of local resection for ampullary tumors, because malignant recurrence of what were thought to be benign tumors has been reported. This study examined the role of local resection in the management of ampullary tumors. Thirty patients (mean age 66 years) had transduodenal local resections performed at UCSF-Moffitt Hospital or the San Francisco VA Medical Center (February, 1992 to March, 2004). Mean follow-up time was 5.8 years. Preoperative biopsies (obtained in all patients) showed 18 adenomas, four adenomas with dysplasia, five adenomas with atypia, one adenoma with dysplasia and focal adenocarcinoma, and two tumors seen on endoscopy, whose biopsies showed only duodenal mucosa. In comparison with the final pathology findings, the results of frozen section examinations for malignancy in 20 patients, during the operation, were false-negative in three cases. The final pathologic diagnosis was 23 villous adenomas, six adenocarcinomas, and one paraganglioma. On preoperative biopsies, all patients who had high-grade dysplasia and one of five patients with atypia turned out to have invasive adenocarcinoma when the entire specimen was examined postoperatively. Two (33%) adenocarcinomas recurred at a mean of 4 years; both had negative margins at the initial resection. Among the 23 adenomas, three (13%) recurred (all as adenomas) at a mean of 3.2 years; in only one of these cases was the margin positive at the time of resection. Tumor size did not influence recurrence rate. Ampullary tumors with high-grade dysplasia on preoperative biopsy should be treated by pancreaticoduodenectomy because they usually harbor malignancy. Recurrence is too common and unpredictable after local resection of malignant lesions for this to be considered an acceptable alternative to pancreaticoduodenectomy. Ampullary adenomas can be resected locally with good results, but the recurrence rate was 13%, so endoscopic surveillance is indicated

  10. Quantum Locality in Game Strategy

    PubMed Central

    Melo-Luna, Carlos A.; Susa, Cristian E.; Ducuara, Andrés F.; Barreiro, Astrid; Reina, John H.

    2017-01-01

    Game theory is a well established branch of mathematics whose formalism has a vast range of applications from the social sciences, biology, to economics. Motivated by quantum information science, there has been a leap in the formulation of novel game strategies that lead to new (quantum Nash) equilibrium points whereby players in some classical games are always outperformed if sharing and processing joint information ruled by the laws of quantum physics is allowed. We show that, for a bipartite non zero-sum game, input local quantum correlations, and separable states in particular, suffice to achieve an advantage over any strategy that uses classical resources, thus dispensing with quantum nonlocality, entanglement, or even discord between the players’ input states. This highlights the remarkable key role played by pure quantum coherence at powering some protocols. Finally, we propose an experiment that uses separable states and basic photon interferometry to demonstrate the locally-correlated quantum advantage. PMID:28327567

  11. Acoustic Localization with Infrasonic Signals

    NASA Astrophysics Data System (ADS)

    Threatt, Arnesha; Elbing, Brian

    2015-11-01

    Numerous geophysical and anthropogenic events emit infrasonic frequencies (<20 Hz), including volcanoes, hurricanes, wind turbines and tornadoes. These sounds, which cannot be heard by the human ear, can be detected from large distances (in excess of 100 miles) due to low frequency acoustic signals having a very low decay rate in the atmosphere. Thus infrasound could be used for long-range, passive monitoring and detection of these events. An array of microphones separated by known distances can be used to locate a given source, which is known as acoustic localization. However, acoustic localization with infrasound is particularly challenging due to contamination from other signals, sensitivity to wind noise and producing a trusted source for system development. The objective of the current work is to create an infrasonic source using a propane torch wand or a subwoofer and locate the source using multiple infrasonic microphones. This presentation will present preliminary results from various microphone configurations used to locate the source.

  12. Thermometry considerations in localized hyperthermia.

    PubMed

    Cetas, T C; Connor, W G

    1978-01-01

    The introduction of local hyperthermia as a method of cancer therapy implies the necessity of quantitative measurements of the thermal dose. Our intention is to describe the nature of the problem, both physically and physiologically, with illustrations drawn from thermographic measurements in phantoms and in animals. The characteristics of a thermometry calibration facility are described. Some measurement problems associated with conventional thermometer probes are mentioned and several new thermometers which were developed for use in the electromagnetic fields are reviewed. We present some of the concepts that will guide the development of noninvasive thermometry. Systemic hyperthermia is not considered. We recommend that other reviews specifically directed toward localized hyperthermia be prepared on the methods of heating and on thermal physiological problems.

  13. Localization in Open Quantum Systems

    NASA Astrophysics Data System (ADS)

    Yusipov, I.; Laptyeva, T.; Denisov, S.; Ivanchenko, M.

    2017-02-01

    In an isolated single-particle quantum system, a spatial disorder can induce Anderson localization. Being a result of interference, this phenomenon is expected to be fragile in the face of dissipation. Here we show that a proper dissipation can drive a disordered system into a steady state with tunable localization properties. This can be achieved with a set of identical dissipative operators, each one acting nontrivially on a pair of sites. Operators are parametrized by a uniform phase, which controls the selection of Anderson modes contributing to the state. On the microscopic level, quantum trajectories of a system in the asymptotic regime exhibit intermittent dynamics consisting of long-time sticking events near selected modes interrupted by intermode jumps.

  14. Quantum Locality in Game Strategy

    NASA Astrophysics Data System (ADS)

    Melo-Luna, Carlos A.; Susa, Cristian E.; Ducuara, Andrés F.; Barreiro, Astrid; Reina, John H.

    2017-03-01

    Game theory is a well established branch of mathematics whose formalism has a vast range of applications from the social sciences, biology, to economics. Motivated by quantum information science, there has been a leap in the formulation of novel game strategies that lead to new (quantum Nash) equilibrium points whereby players in some classical games are always outperformed if sharing and processing joint information ruled by the laws of quantum physics is allowed. We show that, for a bipartite non zero-sum game, input local quantum correlations, and separable states in particular, suffice to achieve an advantage over any strategy that uses classical resources, thus dispensing with quantum nonlocality, entanglement, or even discord between the players’ input states. This highlights the remarkable key role played by pure quantum coherence at powering some protocols. Finally, we propose an experiment that uses separable states and basic photon interferometry to demonstrate the locally-correlated quantum advantage.

  15. Subcellular localization of pituitary enzymes

    NASA Technical Reports Server (NTRS)

    Smith, R. E.

    1970-01-01

    A cytochemical procedure is reported for identifying subcellular sites of enzymes hydrolyzing beta-naphthylamine substrates, and to study the sites of reaction product localization in cells of various tissues. Investigations using the substrate Leu 4-methoxy-8-naphthylamine, a capture with hexonium pararosaniline, and the final chelation of osmium have identified the hydrolyzing enzyme of rat liver cells; this enzyme localized on cell membranes with intense deposition in the areas of the parcanaliculi. The study of cells in the anterior pituitary of the rat showed the deposition of reaction product on cell membrane; and on the membranes of secretion granules contained within the cell. The deposition of reaction product on the cell membrane however showed no increase or decrease with changes in the physiological state of the gland and release of secretion granules from specific cells.

  16. Non-Local Euclidean Medians.

    PubMed

    Chaudhury, Kunal N; Singer, Amit

    2012-11-01

    In this letter, we note that the denoising performance of Non-Local Means (NLM) can be improved at large noise levels by replacing the mean by the Euclidean median. We call this new denoising algorithm the Non-Local Euclidean Medians (NLEM). At the heart of NLEM is the observation that the median is more robust to outliers than the mean. In particular, we provide a simple geometric insight that explains why NLEM performs better than NLM in the vicinity of edges, particularly at large noise levels. NLEM can be efficiently implemented using iteratively reweighted least squares, and its computational complexity is comparable to that of NLM. We provide some preliminary results to study the proposed algorithm and to compare it with NLM.

  17. Quantum Locality in Game Strategy.

    PubMed

    Melo-Luna, Carlos A; Susa, Cristian E; Ducuara, Andrés F; Barreiro, Astrid; Reina, John H

    2017-03-22

    Game theory is a well established branch of mathematics whose formalism has a vast range of applications from the social sciences, biology, to economics. Motivated by quantum information science, there has been a leap in the formulation of novel game strategies that lead to new (quantum Nash) equilibrium points whereby players in some classical games are always outperformed if sharing and processing joint information ruled by the laws of quantum physics is allowed. We show that, for a bipartite non zero-sum game, input local quantum correlations, and separable states in particular, suffice to achieve an advantage over any strategy that uses classical resources, thus dispensing with quantum nonlocality, entanglement, or even discord between the players' input states. This highlights the remarkable key role played by pure quantum coherence at powering some protocols. Finally, we propose an experiment that uses separable states and basic photon interferometry to demonstrate the locally-correlated quantum advantage.

  18. Localization of electrons and excitations

    NASA Astrophysics Data System (ADS)

    Larsson, Sven

    2006-07-01

    Electrons, electron holes, or excitations in finite or infinite 'multimer systems' may be localized or delocalized. In the theory of Hush, localization depends on the ratio Δ/ λ ( Δ/2 = coupling; λ = reorganization energy). The latter theory has been extended to the infinite system [S. Larsson, A. Klimkāns, Mol. Cryst. Liq. Cryst. 355 (2000) 217]. The metal/insulator transition often takes place abruptly as a function of Δ/ λ. It is argued that localization in a system with un-filled bands cannot be determined on the basis of Mott-Hubbard U alone, but depends on the number of accessible valence states, reorganization energy λ and coupling Δ (=2t). In fact U = 0 does not necessarily imply delocalization. The analysis here shows that there are many different situations for an insulator to metal transition. Charge transfer in doped NiO is characterized by Ni 2+ - Ni 3+ exchange while charge transfer in pure NiO is characterized by a disproportionation 2Ni 2+ → Ni + + Ni 3+. In spite of the great differences between these two cases, U has been applied without discrimination to both. The relevant localization parameters appear to be Δ and λ in the first case, with only two oxidation states, and U, Δ and λ in the second case with three oxidation states. The analysis is extended to insulator-metal transitions, giant magnetic resistance (GMR) and high Tc superconductivity (SC). λ and Δ can be determined quite accurately in quantum mechanical calculations involving only one and two monomers, respectively.

  19. Wireless local area network security.

    PubMed

    Bergeron, Bryan P

    2004-01-01

    Wireless local area networks (WLANs) are increasingly popular in clinical settings because they facilitate the use of wireless PDAs, laptops, and other pervasive computing devices at the point of care. However, because of the relative immaturity of wireless network technology and evolving standards, WLANs, if improperly configured, can present significant security risks. Understanding the security limitations of the technology and available fixes can help minimize the risks of clinical data loss and maintain compliance with HIPAA guidelines.

  20. Optimizing drugs for local delivery.

    PubMed

    Collingwood, S; Lock, R; Searcey, M

    2009-12-01

    An international panel of speakers together with approximately 70 delegates were brought together by The Society for Medicines Research's symposium on Optimising Drugs for Local Delivery, held on June 11, 2009 at the Novartis Institutes for Biomedical Research, Horsham, UK. The focus of the conference was on the delivery of drugs direct to the site of action and the consequences of this delivery route on delivery technologies, formulation science and molecular design.

  1. Local Area Network Implementation Plan.

    DTIC Science & Technology

    1985-11-01

    vendor support. The introduction of Ethernets should be taken into account during the procurement of new computing equipment (minicomputers. personal ...eventual connection of the IPD mainframes and two additional types of minicomputers as well as some personal computers. word processing systems, and...connection of personal computers to -" the network. and (6) the connection of facsimile devices to the network. 10 2. LOCAL NETWORKING In this section we

  2. Combining global and local approximations

    NASA Technical Reports Server (NTRS)

    Haftka, Raphael T.

    1991-01-01

    A method based on a linear approximation to a scaling factor, designated the 'global-local approximation' (GLA) method, is presented and shown capable of extending the range of usefulness of derivative-based approximations to a more refined model. The GLA approach refines the conventional scaling factor by means of a linearly varying, rather than constant, scaling factor. The capabilities of the method are demonstrated for a simple beam example with a crude and more refined FEM model.

  3. Combining global and local approximations

    SciTech Connect

    Haftka, R.T. )

    1991-09-01

    A method based on a linear approximation to a scaling factor, designated the 'global-local approximation' (GLA) method, is presented and shown capable of extending the range of usefulness of derivative-based approximations to a more refined model. The GLA approach refines the conventional scaling factor by means of a linearly varying, rather than constant, scaling factor. The capabilities of the method are demonstrated for a simple beam example with a crude and more refined FEM model. 6 refs.

  4. Working within local funding trends.

    PubMed

    Pomales-Connors, Irma

    2004-06-01

    Like politics, environmentalism, and fashion, there are trends in health care research and funding. According to a series of reports by the Foundation Center-which collects, organizes, and communicates information on U.S. philanthropy-it is important to understand the significant financial and programmatic changes in the way foundations give. For pharmacists considering soliciting grant support, it is critical that they become aware of these trends and be responsive to the local or regional environments that affect funding.

  5. Local friction in polyolefin blends

    NASA Astrophysics Data System (ADS)

    Luettmer-Strathmann, Jutta

    2005-07-01

    Processes on different length scales affect the dynamics of chain molecules. The friction experienced by a short chain segment depends on both small-scale chain properties and on the local environment of the segment. As a consequence, the (monomeric) friction coefficients of the two components of a binary polymer blend will, in general, differ from each other and from the friction coefficients of the corresponding melts. In this work, we investigate local friction in polyolefin blends with the aid of a small-scale simulation approach. The polymer chains, in united atom representation, are assumed to occupy the sites of a partially filled simple cubic lattice. The simulation focuses on short chain sections with straight backbones and enumerates all possible binary contacts and relative movements of such sections. By evaluating the exact enumeration results in conjunction with equations of state for the blends, we are able to make predictions about the variation of the friction coefficients with local chain architecture and thermodynamic state (temperature, pressure, and composition). We calculate relative values of friction coefficients at temperatures well above the glass transition for blends of PEP, an alternating copolymer of polyethylene and polypropylene, with polyethylene and polyisobutylene and for blends of polyethylene and atactic polypropylene. We also investigate a blend of PEP with head-to-head polypropylene and compare our results with experimental data.

  6. Local equilibrium in bird flocks

    NASA Astrophysics Data System (ADS)

    Mora, Thierry; Walczak, Aleksandra M.; Del Castello, Lorenzo; Ginelli, Francesco; Melillo, Stefania; Parisi, Leonardo; Viale, Massimiliano; Cavagna, Andrea; Giardina, Irene

    2016-12-01

    The correlated motion of flocks is an example of global order emerging from local interactions. An essential difference with respect to analogous ferromagnetic systems is that flocks are active: animals move relative to each other, dynamically rearranging their interaction network. This non-equilibrium characteristic has been studied theoretically, but its impact on actual animal groups remains to be fully explored experimentally. Here, we introduce a novel dynamical inference technique, based on the principle of maximum entropy, which accommodates network rearrangements and overcomes the problem of slow experimental sampling rates. We use this method to infer the strength and range of alignment forces from data of starling flocks. We find that local bird alignment occurs on a much faster timescale than neighbour rearrangement. Accordingly, equilibrium inference, which assumes a fixed interaction network, gives results consistent with dynamical inference. We conclude that bird orientations are in a state of local quasi-equilibrium over the interaction length scale, providing firm ground for the applicability of statistical physics in certain active systems.

  7. Non-local Optical Potentials

    NASA Astrophysics Data System (ADS)

    Thompson, Ian

    2010-11-01

    In all direct reactions to probe the structure of exotic nuclei at FRIB, optical potentials will be needed in the entrance and exit channels. At high energies Glauber approximations may be useful, but a low energies (5 to 20 MeV/nucleon) other approaches are required. Recent work of the UNEDF project [1] has shown that reaction cross sections at these energies can be accounted for by calculating all inelastic and transfer channels reachable by one particle-hole transitions from the elastic channel. In this model space, we may also calculate the two-step dynamic polarization potential (DPP) that adds to the bare folded potential to form the complex optical potential. Our calculations of the DPP, however, show that its non-localities are very significant, as well as the partial-wave dependence of both its real and imaginary components. The Perey factors (the wave function ratio to that from an equivalent local potential) are more than 20% different from unity, especially for partial waves inside grazing. These factors combine to suggest a reexamination of the validity of local and L-independent fitted optical potentials, especially for capture reactions that are dominated by low partial waves. Prepared by LLNL under Contract DE-AC52-07NA27344. [1] G.P.A. Nobre, F.S. Dietrich, J.E. Escher, I.J. Thompson, M. Dupuis, J. Terasaki and J. Engel, submitted to Phys. Rev. Letts., 2010.

  8. Local-scale dynamics and local drivers of bushmeat trade.

    PubMed

    Nyaki, Angela; Gray, Steven A; Lepczyk, Christopher A; Skibins, Jeffrey C; Rentsch, Dennis

    2014-10-01

    Bushmeat management policies are often developed outside the communities in which they are to be implemented. These policies are also routinely designed to be applied uniformly across communities with little regard for variation in social or ecological conditions. We used fuzzy-logic cognitive mapping, a form of participatory modeling, to compare the assumptions driving externally generated bushmeat management policies with perceptions of bushmeat trade dynamics collected from local community members who admitted to being recently engaged in bushmeat trading (e.g., hunters, sellers, consumers). Data were collected during 9 workshops in 4 Tanzanian villages bordering Serengeti National Park. Specifically, we evaluated 9 community-generated models for the presence of the central factors that comprise and drive the bushmeat trade and whether or not models included the same core concepts, relationships, and logical chains of reasoning on which bushmeat conservation policies are commonly based. Across local communities, there was agreement about the most central factors important to understanding the bushmeat trade (e.g., animal recruitment, low income, and scarcity of food crops). These matched policy assumptions. However, the factors perceived to drive social-ecological bushmeat trade dynamics were more diverse and varied considerably across communities (e.g., presence or absence of collaborative law enforcement, increasing human population, market demand, cultural preference). Sensitive conservation issues, such as the bushmeat trade, that require cooperation between communities and outside conservation organizations can benefit from participatory modeling approaches that make local-scale dynamics and conservation policy assumptions explicit. Further, communities' and conservation organizations' perceptions need to be aligned. This can improve success by allowing context appropriate policies to be developed, monitored, and appropriately adapted as new evidence is

  9. Dynamical Localization in Molecular Systems.

    NASA Astrophysics Data System (ADS)

    Wang, Xidi

    In the first four chapters of this thesis we concentrate on the Davydov model which describes the vibrational energy quanta of Amide I bonds (C=O bonds on the alpha -helix) coupled to the acoustic phonon modes of the alpha-helix backbone in the form of a Frohlich Hamiltonian. Following a brief introduction in chapter one, in chapter two we formulate the dynamics of vibrational quanta at finite temperature by using coherent state products. The fluctuation-dissipation relation is derived. At zero temperature, in the continuum limit, we recover the original results of Davydov. We also achieve good agreement with numerical simulations. In chapter three, the net contraction of the lattice is calculated exactly at any temperature, and its relation to the so -call "topological stability" of the Davydov soliton is discussed. In the second section of the chapter three we calculate the overtone spectra of crystalline acetanilide (according to some opinions ACN provides experimental evidence for the existence of Davydov solitons). Good agreement with experimental data has been obtained. In chapter four we study the self-trapped vibrational excitations by the Quantum Monte Carlo technique. For a single excitation, the temperature dependence of different physical observables is calculated. The quasi-particle which resembles the Davydov soliton has been found to be fairly narrow using the most commonly used data for the alpha -helix; at temperatures above a few Kelvin, the quasi-particle reaches its smallest limit (extends over three sites), which implies diffusive motion of the small polaron-like quasi-particle at high temperatures. For the multi-excitation case, bound pairs and clusters of excitations are found at low temperatures; they gradually dissociate when the temperature of the system is increased as calculated from the density-density correlation function. In the last chapter of this thesis, we study a more general model of dynamical local modes in molecular systems

  10. Allergy to local anesthetics: Reality or myth?

    PubMed

    Malinovsky, Jean-Marc; Chiriac, Anca M; Tacquard, Charles; Mertes, Paul Michel; Demoly, Pascal

    2016-09-01

    The incidence of allergic reactions to local anesthetics is low. Most cases involve a psychogenic reaction rather than an allergic reaction. Additives and preservatives added to local anesthetics may cause allergic reactions. Vascular resorption of epinephrine-containing local anesthetics may produce cardiovascular signs similar to an allergic reaction. Diagnosis of allergy to local anesthetics must be established by skin testing and provocative challenge.

  11. Computational methods for global/local analysis

    NASA Technical Reports Server (NTRS)

    Ransom, Jonathan B.; Mccleary, Susan L.; Aminpour, Mohammad A.; Knight, Norman F., Jr.

    1992-01-01

    Computational methods for global/local analysis of structures which include both uncoupled and coupled methods are described. In addition, global/local analysis methodology for automatic refinement of incompatible global and local finite element models is developed. Representative structural analysis problems are presented to demonstrate the global/local analysis methods.

  12. A Century of Local History Writing.

    ERIC Educational Resources Information Center

    Howe, Barbara J.

    1989-01-01

    Offers suggestions on finding sources on the historiography of local history. Examines books useful to the local historian, including books on local social history and studies on immigrants, minorities, and women. Suggests the use of visual versions of local history. Discusses architectural histories, landmarks, and the use of primary sources. (RW)

  13. [Symptoms. Localizations: knee, hip, hands, spine, other localizations].

    PubMed

    Pérez Martín, Álvaro

    2014-01-01

    The symptoms of osteoarthritis vary widely from patient to patient, depending especially on the localization on the disease. There is a poor correlation between radiological involvement and pain. In general, symptom onset is gradual and symptoms increase slowly but progressively. The most commonly affected joints are the knees, hips, hands, and spine. The main signs and symptoms are pain, stiffness, joint deformity, and crepitus. Pain is mechanical and its causes are multifactorial; in the initial phases, pain usually manifests in self-limiting episodes but may subsequently be almost constant. The criteria of the American college of Rheumatology for the classification of osteoarthritis of the knee, hip and hands are an aid to classification and standardization but are not useful for diagnosis. Hip osteoarthritis usually produces inguinal pain in the internal and anterior sections of the muscle extending to the knee and, with progression, tends to limit mobility. Knee osteoarthritis is more frequent in women and is usually associated with hand osteoarthritis and obesity. In hand osteoarthritis, the most commonly affected joints are the distal interphalangeal joints, followed by the proximal interphalangeal joints and the trapeziometacarpal joints; the development of Heberden and Bouchard nodes is common; involvement of the trapeziometacarpal joint is called rhizarthrosis and is one of the forms of osteoarthritis that produces the greatest limitation on hand function. Osteoarthritis of the spine affects the facet joints and the vertebral bodies. Other, less frequent, localizations are the foot, elbow and shoulder, which are generally secondary forms of osteoarthritis.

  14. Local and non-local correlations in nanoscopic systems

    NASA Astrophysics Data System (ADS)

    Sangiovanni, Giorgio; Valli, A.; Rohringer, G.; Toschi, A.; Held, K.; Das, H.; Saha-Dasgupta, T.

    2012-02-01

    Tools for reliably treating nanoscopic systems, like coupled quantum-dots, ad-atoms on surfaces, macromolecules, etc., in the presence of electronic correlations are either missing or prohibitively expensive. We have implemented a new computational scheme based on a self-consistently defined set of local problems [1]. Our method scales linearly with the number of sites and allows us to perform large-scale sign-problem free Quantum Monte-Carlo simulations. We have studied the behavior of a single-atom junction formed upon stretching a metallic wire and found that a metal-insulator crossover is induced when the wire is about to break up. The combination with ab-initio techniques allowed us to study size-dependent properties of Manganite nano-clusters [2]. The simplest implementation of our method includes only local self-energy effects. We recently went beyond this and applied the resulting more sophisticated version of our method to an exactly solvable model finding results in remarkable agreement with the exact solution. [1] A. Valli, G. Sangiovanni, O. Gunnarsson, A. Toschi and K. Held, PRL 104, 246402 (2010) [2] H. Das, G. Sangiovanni, A. Valli, K. Held and T. Saha-Dasgupta, PRL 107, 197202 (2011)

  15. The Spitzer Local Volume Legacy

    NASA Astrophysics Data System (ADS)

    Kennicutt, Robert; Lee, J. C.; Engelbracht, C.; Begum, A.; Block, M.; Calzetti, D.; Dalcanton, J.; Dale, D.; Funes, J.; Gil de Paz, A.; Gordon, K.; Johnson, B.; Sakai, S.; Skillman, E.; van Zee, L.; Walter, F.; Weisz, D.; Williams, B.; Wu, Y.

    2007-12-01

    The Local Volume Legacy (LVL) is a Spitzer Cycle 4 Legacy project, aimed at obtaining IRAC and MIPS imaging for a complete sample of 258 galaxies within 11 Mpc. Our observations probe the spatially- resolved star formation, dust, and red stellar populations of galaxies that have been drawn from a statistically robust local sample, in which a full diversity of galaxy properties such as luminosities, surface brightnesses, metallicities are represented. Our sample includes: (i) a complete volume-limited galaxy sample within 3.5 Mpc, and (ii) an unbiased sample of S-Irr galaxies within an 11 Mpc sphere. LVL will produce a multi-wavelength census of the Galactic neighborhood, extending to the faintest limits of the galactic luminosity function and exploiting the highest spatial resolution and absolute depth achievable with Spitzer. Our ancillary dataset includes H-alpha and UV imaging from the GALEX 11HUGS and NGS surveys, stellar population mapping from the HST ANGST Treasury survey, HI mapping with the VLA and GMRT, and optical broad-band imaging and spectroscopy. By homogeneously filling in critical gaps in the current Spitzer coverage of the Local Volume, and providing SED coverage from the UV to the FIR, LVL will supply an enduring homogeneous core dataset on the Galactic neighborhood for the astronomical community. Science issues to be addressed include: constraining the physical mechanisms underlying dust heating and understanding correlations between FIR emission, dust content and global galaxy properties; establishing the primary factors which influence PAH emission and evaluating the robustness of PAH emission as a SFR indicator, particularly at low metallicities and high specific SFRs; probing the temporal variation of star formation as a function of global properties, with special focus on dwarf galaxies. This poster will highlight the scientific goals and design of the survey, and present early results from the imaging campaign.

  16. On the local stellar populations

    NASA Astrophysics Data System (ADS)

    Fuhrmann, Klaus; Chini, Rolf; Kaderhandt, Lena; Chen, Zhiwei

    2017-01-01

    We present a study of the local stellar populations from a volume-complete all-sky survey of the about 500 bright stars with distances less than 25 pc and down to main-sequence effective temperatures Teff ≥ 5300 K. The sample is dominated by a 93 per cent fraction of Population I stars, only 22 sources (5 per cent) are Population II stars, and 9 sources (2 per cent) are intermediate-disc stars. No source belongs to the halo. By following the mass of the stars instead of their light, the resulting subset of 136 long-lived stars distributes as 22 (16.2 per cent):6 (4.4 per cent):108 (79.4 per cent) for the Population II:intermediate disc:Population I, respectively. Along with the much larger scaleheight reached by Population II, this unbiased census of long-lived stars provides plain evidence for a starburst epoch in the early Milky Way, with the formation of a massive, rotationally supported, and dark Population II. The same conclusion arises from the substantial early chemical enrichment levels, exemplified here by the elements magnesium and iron, as it arises also from the local Population II white dwarfs. The kinematics, metallicity distribution functions, star formation rates, age-metallicity relations, the inventory of young stars, and the occurrence of blue straggler stars are discussed. A potentially new aspect of the survey is the possibility for substructure among the local Population II stars that may further subdivide into metal-poor and metal-rich sources.

  17. Genetic Algorithms and Local Search

    NASA Technical Reports Server (NTRS)

    Whitley, Darrell

    1996-01-01

    The first part of this presentation is a tutorial level introduction to the principles of genetic search and models of simple genetic algorithms. The second half covers the combination of genetic algorithms with local search methods to produce hybrid genetic algorithms. Hybrid algorithms can be modeled within the existing theoretical framework developed for simple genetic algorithms. An application of a hybrid to geometric model matching is given. The hybrid algorithm yields results that improve on the current state-of-the-art for this problem.

  18. State and Local Government Partnership

    SciTech Connect

    Barton, Alexander; Rinebold, Joel; Aresta, Paul

    2012-03-30

    The State and Local Government Partnership project has built relationships between the Department of Energy (DOE), regional states, and municipalities. CCAT implemented this project using a structure that included leadership by the DOE. Outreach was undertaken through collaborative meetings, workshops, and briefings; the development of technical models and local energy plans; support for state stakeholder groups; and implementation of strategies to facilitate the deployment of hydrogen and fuel cell technologies. The final guidance documents provided to stakeholders consisted of individual strategic state “Roadmaps” to serve as development plans. These “Roadmaps” confirm economic impacts, identify deployment targets, and compare policies and incentives for facility development in each of the regional states. The partnerships developed through this project have improved the exchange of knowledge between state and local government stakeholders and is expected to increase the deployment of hydrogen and fuel cell technologies in early market applications, consistent with the DOE’s market transformation efforts. Technically accurate and objective information was, and continues to be, provided to improve public and stakeholder perceptions regarding the use of hydrogen and fuel cell technologies. Based on the “Roadmaps” and studies conducted for this project, there is the potential to generate approximately 10.75 million megawatt hours (MWh) of electricity annually from hydrogen and fuel cell technologies at potential host sites in the Northeast regional states, through the development of 1,364 to 1,818 megawatts (MW) of fuel cell electric generation capacity. Currently, the region has approximately 1,180 companies that are part of the growing hydrogen and fuel cell industry supply chain in the region. These companies are estimated to have over $1 billion in annual revenue and investment, contribute more than $51 million in annual state and local tax revenue

  19. Multi-directional local search.

    PubMed

    Tricoire, Fabien

    2012-12-01

    This paper introduces multi-directional local search, a metaheuristic for multi-objective optimization. We first motivate the method and present an algorithmic framework for it. We then apply it to several known multi-objective problems such as the multi-objective multi-dimensional knapsack problem, the bi-objective set packing problem and the bi-objective orienteering problem. Experimental results show that our method systematically provides solution sets of comparable quality with state-of-the-art methods applied to benchmark instances of these problems, within reasonable CPU effort. We conclude that the proposed algorithmic framework is a viable option when solving multi-objective optimization problems.

  20. Localization of hidden Chua's attractors

    NASA Astrophysics Data System (ADS)

    Leonov, G. A.; Kuznetsov, N. V.; Vagaitsev, V. I.

    2011-06-01

    The classical attractors of Lorenz, Rossler, Chua, Chen, and other widely-known attractors are those excited from unstable equilibria. From computational point of view this allows one to use numerical method, in which after transient process a trajectory, started from a point of unstable manifold in the neighborhood of equilibrium, reaches an attractor and identifies it. However there are attractors of another type: hidden attractors, a basin of attraction of which does not contain neighborhoods of equilibria. In the present Letter for localization of hidden attractors of Chua's circuit it is suggested to use a special analytical-numerical algorithm.

  1. A Localized Ensemble Kalman Smoother

    NASA Technical Reports Server (NTRS)

    Butala, Mark D.

    2012-01-01

    Numerous geophysical inverse problems prove difficult because the available measurements are indirectly related to the underlying unknown dynamic state and the physics governing the system may involve imperfect models or unobserved parameters. Data assimilation addresses these difficulties by combining the measurements and physical knowledge. The main challenge in such problems usually involves their high dimensionality and the standard statistical methods prove computationally intractable. This paper develops and addresses the theoretical convergence of a new high-dimensional Monte-Carlo approach called the localized ensemble Kalman smoother.

  2. Nonclassicality of local bipartite correlations

    NASA Astrophysics Data System (ADS)

    Jebaratnam, C.; Aravinda, S.; Srikanth, R.

    2017-03-01

    Simulating quantum nonlocality and steering requires augmenting preshared randomness with nonvanishing communication cost. This prompts the question of how one may provide such an operational characterization for the quantumness of correlations due to even unentangled states. Here we show that for a certain class of states, such quantumness can be pointed out by superlocality, the requirement for a larger dimension of the preshared randomness to simulate the correlations than that of the quantum state that generates them. This provides an approach to define the nonclassicality of local multipartite correlations in convex operational theories.

  3. [Carotid endarterectomy under local anesthesia].

    PubMed

    Kuz'min, A L; Belov, Iu V

    2001-01-01

    Results of carotid endarterectomy (CEAE) in 193 patients with different degree of cerebrovascular insufficiency were analyzed. All the patients were men with carotid atherosclerosis (age from 39 to 68 years, mean age 53.6 +/- 0.4). A total of 253 CEAEs were performed under local anesthesia (60 patients underwent consecutive bilateral operations). In early postoperative period 3 patients died, one of them--of ischemic stroke due to thrombosis of internal carotid artery on the side of the operation. Non-fatal stroke was in 1 patient. There were no intraoperative cerebral complications. This testifies to reliability of cerebral circulation control through direct contact with patient.

  4. Exploring Mars using local resources

    NASA Astrophysics Data System (ADS)

    Brody, Steven; McKay, Christopher; Kaplan, David; Rapp, Donald

    1996-03-01

    This paper takes the reader on a second human expedition to Mars. Following confirmation that life once existed on Mars during the first human mission launched in 2018, crew members of our subsequent mission, assisted by robotic devices, spend two years on Mars pursuing three goals: (1) continue the search for further evidence of ancient life and what may still exist, (2) safely endure their extended stay on Mars, and (3) lay the foundation for the eventual human settlement of the planet. A cornerstone in achieving these three goals, as discussed in the paper, will be the utilization of local resources as we further learn to ``live off the land''.

  5. Strabismus complications from local anesthetics.

    PubMed

    Guyton, David L

    2008-01-01

    Strabismus developing after retrobulbar or peribulbar anesthesia for both anterior and posterior segment eye surgery may be due to myotoxicity to an extraocular muscle from the local anesthetic agent. Initial paresis often causes diplopia immediately after surgery, but later progressive segmental fibrosis occurs, and/or hypertrophy of the muscle, producing diplopia in the opposite direction from the direction of the initial diplopia. The inferior rectus muscle is most commonly affected. Usually a large recession on an adjustable suture of the involved muscle(s) yields good alignment. Using topical anesthesia or sub-Tenon's anesthesia can avoid this complication.

  6. A local Vapnik-Chervonenkis complexity.

    PubMed

    Oneto, Luca; Anguita, Davide; Ridella, Sandro

    2016-10-01

    We define in this work a new localized version of a Vapnik-Chervonenkis (VC) complexity, namely the Local VC-Entropy, and, building on this new complexity, we derive a new generalization bound for binary classifiers. The Local VC-Entropy-based bound improves on the original Vapnik's results because it is able to discard those functions that, most likely, will not be selected during the learning phase. The result is achieved by applying the localization principle to the original global complexity measure, in the same spirit of the Local Rademacher Complexity. By exploiting and improving a recently developed geometrical framework, we show that it is also possible to relate the Local VC-Entropy to the Local Rademacher Complexity by finding an admissible range for one given the other. In addition, the Local VC-Entropy allows one to reduce the computational requirements that arise when dealing with the Local Rademacher Complexity in binary classification problems.

  7. Local entropy of a nonequilibrium fermion system

    NASA Astrophysics Data System (ADS)

    Stafford, Charles A.; Shastry, Abhay

    2017-03-01

    The local entropy of a nonequilibrium system of independent fermions is investigated and analyzed in the context of the laws of thermodynamics. It is shown that the local temperature and chemical potential can only be expressed in terms of derivatives of the local entropy for linear deviations from local equilibrium. The first law of thermodynamics is shown to lead to an inequality, not equality, for the change in the local entropy as the nonequilibrium state of the system is changed. The maximum entropy principle (second law of thermodynamics) is proven: a nonequilibrium distribution has a local entropy less than or equal to a local equilibrium distribution satisfying the same constraints. It is shown that the local entropy of the system tends to zero when the local temperature tends to zero, consistent with the third law of thermodynamics.

  8. Local dimensionality determines imaging speed in localization microscopy

    PubMed Central

    Fox-Roberts, Patrick; Marsh, Richard; Pfisterer, Karin; Jayo, Asier; Parsons, Maddy; Cox, Susan

    2017-01-01

    Localization microscopy allows biological samples to be imaged at a length scale of tens of nanometres. Live-cell super-resolution imaging is rare, as it is generally assumed to be too slow for dynamic samples. The speed of data acquisition can be optimized by tuning the density of activated fluorophores in each time frame. Here, we show that the maximum achievable imaging speed for a particular structure varies by orders of magnitude, depending on the sample dimensionality (that is, whether the sample is more like a point, a strand or an extended structure such as a focal adhesion). If too high an excitation density is used, we demonstrate that the analysis undergoes silent failure, resulting in reconstruction artefacts. We are releasing a tool to allow users to identify areas of the image in which the activation density was too high and correct for them, in both live- and fixed-cell experiments. PMID:28079054

  9. Local dimensionality determines imaging speed in localization microscopy

    NASA Astrophysics Data System (ADS)

    Fox-Roberts, Patrick; Marsh, Richard; Pfisterer, Karin; Jayo, Asier; Parsons, Maddy; Cox, Susan

    2017-01-01

    Localization microscopy allows biological samples to be imaged at a length scale of tens of nanometres. Live-cell super-resolution imaging is rare, as it is generally assumed to be too slow for dynamic samples. The speed of data acquisition can be optimized by tuning the density of activated fluorophores in each time frame. Here, we show that the maximum achievable imaging speed for a particular structure varies by orders of magnitude, depending on the sample dimensionality (that is, whether the sample is more like a point, a strand or an extended structure such as a focal adhesion). If too high an excitation density is used, we demonstrate that the analysis undergoes silent failure, resulting in reconstruction artefacts. We are releasing a tool to allow users to identify areas of the image in which the activation density was too high and correct for them, in both live- and fixed-cell experiments.

  10. Developing Photo Activated Localization Microscopy

    NASA Astrophysics Data System (ADS)

    Hess, Harald

    2015-03-01

    Photo Activated Localization Microscopy, PALM, acquires super-resolution images by activating a subset of activatable fluorescent labels and estimating the center of the each molecular label to sub-diffractive accuracy. When this process is repeated thousands of times for different subsets of molecules, then an image can be rendered from all the center coordinates of the molecules. I will describe the circuitous story of its development that began with another super-resolution technique, NSOM, developed by my colleague Eric Betzig, who imaged single molecules at room temperature, and later we spectrally resolved individual luminescent centers of quantum wells. These two observations inspired a generalized path to localization microscopy, but that path was abandoned because no really useful fluorescent labels were available. After a decade of nonacademic industrial pursuits and the subsequent freedom of unemployment, we came across a class of genetically expressible fluorescent proteins that were switchable or convertible that enabled the concept to be implemented and be biologically promising. The past ten years have been very active with many groups exploring applications and enhancements of this concept. Demonstrating significant biological relevance will be the metric if its success.

  11. ISS Local Environment Spectrometers (ISLES)

    NASA Technical Reports Server (NTRS)

    Krause, Linda Habash; Gilchrist, Brian E.

    2014-01-01

    In order to study the complex interactions between the space environment surrounding the ISS and the ISS surface materials, we propose to use lowcost, high-TRL plasma sensors on the ISS robotic arm to probe the ISS space environment. During many years of ISS operation, we have been able to condut effective (but not perfect) extravehicular activities (both human and robotic) within the perturbed local ISS space environment. Because of the complexity of the interaction between the ISS and the LEO space environment, there remain important questions, such as differential charging at solar panel junctions (the so-called "triple point" between conductor, dielectric, and space plasma), increased chemical contamination due to ISS surface charging and/or thruster activation, water dumps, etc, and "bootstrap" charging of insulating surfaces. Some compelling questions could synergistically draw upon a common sensor suite, which also leverages previous and current MSFC investments. Specific questions address ISS surface charging, plasma contactor plume expansion in a magnetized drifting plasma, and possible localized contamination effects across the ISS.

  12. [The local manifestations of diphtheria].

    PubMed

    Kapustian, V A; Boldyrev, V V; Maleev, V V; Mikhaĭlova, E I; Sedak, E F

    1994-01-01

    The development of diphtheria is characterized by a pronounced local process, but the description of local changes in the disease are based only on postmortem findings. 67 patients with different forms of diphtheria were examined. In 11 cases of descending croup bronchoscopic examination was carried out. As revealed in this study, at the primary stage of the disease films cover the whole of the air duct system, and the process is identical to that on tonsils. In the course of convalescence and under the action of specific antiserum films on the tonsils, the soft palate and the vocal cords disappeared, and films in the larynx, the trachea and bronchi could be easily separated from the mucous membrane, but always with the formation of erosions and even perforations in it. The character of the process was indicative of the absence of any specific action of diphtheria toxin on the mucous membrane at the site of the inoculation of the infective agent. No correlation between the severity of the course of diphtheria, the degree of edema and the frequency of the development of complications was noted.

  13. Local Effects of Delayed Food

    PubMed Central

    Davison, Michael; Baum, William M

    2007-01-01

    Five pigeons were trained on a procedure in which seven concurrent variable-interval schedules arranged seven different food–rate ratios in random sequence in each session. Each of these components lasted for 10 response-produced food deliveries, and components were separated by 10-s blackouts. We varied delays to food (signaled by blackout) between the two response alternatives in an experiment with three phases: In Phase 1, the delay on one alternative was 0 s, and the other was varied between 0 and 8 s; in Phase 2, both delays were equal and were varied from 0 to 4 s; in Phase 3, the two delays summed to 8 s, and each was varied from 1 to 7 s. The results showed that increasing delay affected local choice, measured by a pulse in preference, in the same way as decreasing magnitude, but we found also that increasing the delay at the other alternative increased local preference. This result casts doubt on the traditional view that a reinforcer strengthens a response depending only on the reinforcer's value discounted by any response–reinforcer delay. The results suggest that food guides, rather than strengthens, behavior. PMID:17465314

  14. Local optimization of energy systems

    SciTech Connect

    Lozano, M.A.; Valero, A.; Serra, L.

    1996-12-31

    Many thermal systems are very complex due to the number of components and/or its strong interdependence. This complexity makes difficult the optimization of the system design and operation. The theory of Exergetic Cost is based on concepts such as resources, structure, efficiency and purpose (belonging to any theory of production) and on the Second Law. This paper will show how it is possible to obtain from the theory of exergetic cost the marginal costs (Lagrange multipliers) of local resources being consumed by a component. This paper also shows the advantage of the proposed Theory of Perturbations when describing the complexity of structural interactions in a straightforward way. This theory allows to formulate simple procedures for local optimization of components in a plant. Finally, strategies for optimization of complex systems are shown. They are based in the sequential optimization from component to component. This clear and efficient method comes form the fact that the authors have now an operative application of the Thermoeconomic Isolation Principle. This is applied here to thermal power plants.

  15. Localized vibrational modes in diamond

    NASA Astrophysics Data System (ADS)

    Murzaev, R. T.; Bachurin, D. V.; Korznikova, E. A.; Dmitriev, S. V.

    2017-03-01

    Discrete breather (DB) or, synonymously, intrinsic localized mode (ILM) is a spatially localized and time-periodic vibrational mode in a defect-free nonlinear lattice, e.g., in a crystal lattice. Standing DB and DB clusters (double and triple) are studied in diamond using molecular dynamics method with the AIREBO interatomic potentials. Single DB can be easily excited by applying initial shifts, A0, to a pair of nearest atoms along the valence bond in the opposite directions. Admissible excitation amplitudes are 0.09 ≤A0 /a0 ≤ 0.12, where a0 is the equilibrium interatomic distance. The core of a DB is a pair of nearest carbon atoms oscillating out-of-phase, while the neighboring atoms oscillate with one order of magnitude lower amplitudes. DB frequency is above the top of the phonon spectrum and increases with the oscillation amplitude. DB lives for more than 100 oscillation periods which approximately corresponds to 2 ps. The range of initial amplitudes and other conditions necessary for the excitation of double and triple DB clusters as well as their lifetime are investigated in detail. Two different mechanisms of energy exchange between DBs in the DB clusters are revealed, which is the main result of the present study. Our results contribute to a deeper understanding of the nonlinear lattice dynamics of diamond.

  16. Severe Local Storms Cultural Heritage

    NASA Astrophysics Data System (ADS)

    Gladich, I.; Gallai, I.; Giaiotti, D. B.; Morgan, G. M.; Stel, F.

    2009-09-01

    Local storms always had a deep impact on people communities, mainly because of the severe damage caused, because of their unpredictability and, up to a few years ago, even because of the lack of knowledge and awareness on their physical origin. Because of this large impact on real life and on imagination, people needed and wanted to describe and report the occurrence of these events, giving them suited names. Often, these nouns are related to the myth developed to explain the cause of the events. In this work, a short presentation and description of the popular nouns used to describe severe local storm events in different areas of the World is given. Countries taken into account span from Italy, moving toward Africa and reaching a few communities of Native Americans. The etymology of the names gives interesting information, useful even under the anthropological point of view, on the Culture and Believes of the peoples who adopted them. This research work is the result of an underground activity carried out in the last ten years by the authors, during their contacts with students and researchers coming from different Countries and mainly met at the International Center for Theoretical Physics in Trieste.

  17. Hardy's test for local realism

    NASA Astrophysics Data System (ADS)

    Nguyen, Quynh; Dregli, Aurelio

    2014-03-01

    We demonstrated the violation of Bell inequality using entangled photons produced by spontaneous parametric down-conversion. The experiment is based on a local realism test originally proposed by Lucien Hardy. Polarized entangled photons is produced in down-conversion through a pair of beta barium borate crystals. Polarization is adjusted by half wave plates and quartz plates. Single photons travel down two separate paths, each pass through a half-wave plate and a broad band polarizing beam splitter. Light from the beam splitter is collected by lens and focused into optic fibers that direct light into a single photon counting module. Coincidence counting is done using time-to-amplitude converter/single-channel-analyzer units. The probabilities in Bell-Clauser-Horne inequality is calculated by the ratio of the measured coincidence counts on the transmitted laser beams to the total number of coincidence. The counting is done by a LabVIEW program. We find the data to violate local realism by 30 standard deviation. The experiment is based on straight forward quantum mechanical calculations and experimental techniques more accessible to undergraduate students than other methods of testing Bell inequalities.

  18. Giants in the Local Region

    NASA Astrophysics Data System (ADS)

    Luck, R. Earle; Heiter, Ulrike

    2007-06-01

    We present parameter and abundance data for a sample of 298 nearby giants. The spectroscopic data for this work have a resolution of R~60,000, S/N>150, and spectral coverage from 475 to 685 nm. Overall trends in the Z>10 abundances are dominated by Galactic chemical evolution, while the light-element abundances are influenced by stellar evolution, as well as Galactic evolution. We find several super-Li stars in our sample and confirm that Li abundances in the first giant branch are related to mixing depths. Once astration of lithium on the main sequence along with the overall range of main-sequence lithium abundances are taken into account, the lithium abundances of the giants are not dramatically at odds with the predictions of standard stellar evolution. We find the giants to be carbon-diluted in accord with standard stellar evolution and that the carbon and oxygen abundances determined for the local giants are consistent with those found in local field dwarfs. We find that there is evidence for systematic carbon variations in the red giant clump in the sense that the blue side of the clump is carbon-poor (more diluted) than the red side.

  19. Localizing ventricular tachycardia through entrainment.

    PubMed

    Kuo, C T; Luqman, N; Lin, K H; Chiang, C W

    2000-12-01

    Area(s) of slow conduction are thought to be present within the reentry circuit of most clinically important ventricular tachycardia (VT). To prevent recurrence after ablation of VT late after myocardial infarction, it is desirable to localize and destroy area(s) of slow conduction "critical link" within the reentry circuit. Conventionally, they may be identified by endocardial catheter mapping, continuous electrical activity, mid-diastolic potentials, earliest endocardial activation, pace-mapping etc. However, none of these methods are very specific. Entrainment method may be used to localize the slow conduction zone of reentrant VT. Concealed entrainment is consistent with pacing at a site in the reentry circuit but may also occur at some "bystander" sites that are close to the reentry circuit but are not participating in the circuit itself. During pacing at the slow conduction area of the reentry circuit, the stimulus to QRS (S-QRS) interval should equal the electrogram to QRS (EG-QRS) interval during VT. Similarly the post-pacing interval (PPI) approximates the tachycardia cycle length. During pacing at bystander sites, the S-QRS interval may be greater, less than or equal to the EG-QRS interval, depending on the conduction time from the bystander site to the circuit. The PPI, however, always exceed the tachycardia cycle length. In conjunction with concealed entrainment, the use of diastolic potential, double potentials and continuous electrical activity enhances the prediction of radiofrequency termination of post-infarction VT.

  20. Causal localizations in relativistic quantum mechanics

    SciTech Connect

    Castrigiano, Domenico P. L. Leiseifer, Andreas D.

    2015-07-15

    Causal localizations describe the position of quantum systems moving not faster than light. They are constructed for the systems with finite spinor dimension. At the center of interest are the massive relativistic systems. For every positive mass, there is the sequence of Dirac tensor-localizations, which provides a complete set of inequivalent irreducible causal localizations. They obey the principle of special relativity and are fully Poincaré covariant. The boosters are determined by the causal position operator and the other Poincaré generators. The localization with minimal spinor dimension is the Dirac localization. Thus, the Dirac equation is derived here as a mere consequence of the principle of causality. Moreover, the higher tensor-localizations, not known so far, follow from Dirac’s localization by a simple construction. The probability of localization for positive energy states results to be described by causal positive operator valued (PO-) localizations, which are the traces of the causal localizations on the subspaces of positive energy. These causal Poincaré covariant PO-localizations for every irreducible massive relativistic system were, all the more, not known before. They are shown to be separated. Hence, the positive energy systems can be localized within every open region by a suitable preparation as accurately as desired. Finally, the attempt is made to provide an interpretation of the PO-localization operators within the frame of conventional quantum mechanics attributing an important role to the negative energy states.

  1. Causal localizations in relativistic quantum mechanics

    NASA Astrophysics Data System (ADS)

    Castrigiano, Domenico P. L.; Leiseifer, Andreas D.

    2015-07-01

    Causal localizations describe the position of quantum systems moving not faster than light. They are constructed for the systems with finite spinor dimension. At the center of interest are the massive relativistic systems. For every positive mass, there is the sequence of Dirac tensor-localizations, which provides a complete set of inequivalent irreducible causal localizations. They obey the principle of special relativity and are fully Poincaré covariant. The boosters are determined by the causal position operator and the other Poincaré generators. The localization with minimal spinor dimension is the Dirac localization. Thus, the Dirac equation is derived here as a mere consequence of the principle of causality. Moreover, the higher tensor-localizations, not known so far, follow from Dirac's localization by a simple construction. The probability of localization for positive energy states results to be described by causal positive operator valued (PO-) localizations, which are the traces of the causal localizations on the subspaces of positive energy. These causal Poincaré covariant PO-localizations for every irreducible massive relativistic system were, all the more, not known before. They are shown to be separated. Hence, the positive energy systems can be localized within every open region by a suitable preparation as accurately as desired. Finally, the attempt is made to provide an interpretation of the PO-localization operators within the frame of conventional quantum mechanics attributing an important role to the negative energy states.

  2. Multi-directional local search

    PubMed Central

    Tricoire, Fabien

    2012-01-01

    This paper introduces multi-directional local search, a metaheuristic for multi-objective optimization. We first motivate the method and present an algorithmic framework for it. We then apply it to several known multi-objective problems such as the multi-objective multi-dimensional knapsack problem, the bi-objective set packing problem and the bi-objective orienteering problem. Experimental results show that our method systematically provides solution sets of comparable quality with state-of-the-art methods applied to benchmark instances of these problems, within reasonable CPU effort. We conclude that the proposed algorithmic framework is a viable option when solving multi-objective optimization problems. PMID:25140071

  3. Exploring Mars using local resources

    SciTech Connect

    Brody, S.; McKay, C.; Kaplan, D.; Rapp, D.

    1996-03-01

    This paper takes the reader on a {ital second} human expedition to Mars. Following confirmation that life once existed on Mars during the first human mission launched in 2018, crew members of our subsequent mission, assisted by robotic devices, spend two years on Mars pursuing three goals: (1) continue the search for further evidence of ancient life and what may still exist, (2) safely endure their extended stay on Mars, and (3) lay the foundation for the eventual human settlement of the planet. A cornerstone in achieving these three goals, as discussed in the paper, will be the utilization of local resources as we further learn to {open_quote}{open_quote}live off the land{close_quote}{close_quote}. {copyright} {ital 1996 American Institute of Physics.}

  4. Headphone localization of speech stimuli

    NASA Technical Reports Server (NTRS)

    Begault, Durand R.; Wenzel, Elizabeth M.

    1991-01-01

    Recently, three dimensional acoustic display systems have been developed that synthesize virtual sound sources over headphones based on filtering by Head-Related Transfer Functions (HRTFs), the direction-dependent spectral changes caused primarily by the outer ears. Here, 11 inexperienced subjects judged the apparent spatial location of headphone-presented speech stimuli filtered with non-individualized HRTFs. About half of the subjects 'pulled' their judgements toward either the median or the lateral-vertical planes, and estimates were almost always elevated. Individual differences were pronounced for the distance judgements; 15 to 46 percent of stimuli were heard inside the head with the shortest estimates near the median plane. The results infer that most listeners can obtain useful azimuth information from speech stimuli filtered by nonindividualized RTFs. Measurements of localization error and reversal rates are comparable with a previous study that used broadband noise stimuli.

  5. Locally vascularized pelvic accessory spleen.

    PubMed

    Iorio, F; Frantellizzi, V; Drudi, Francesco M; Maghella, F; Liberatore, M

    2016-01-01

    Polysplenism and accessory spleen are congenital, usually asymptomatic anomalies. A rare case of polysplenism with ectopic spleen in pelvis of a 67-year-old, Caucasian female is reported here. A transvaginal ultrasound found a soft well-defined homogeneous and vascularized mass in the left pelvis. Patient underwent MRI evaluation and contrast-CT abdominal scan: images with parenchymal aspect, similar to spleen were obtained. Abdominal scintigraphy with 99mTc-albumin nanocolloid was performed and pelvic region was studied with planar scans and SPECT. The results showed the presence of an uptake area of the radiopharmaceutical in the pelvis, while the spleen was normally visualized. These findings confirmed the presence of an accessory spleen with an artery originated from the aorta and a vein that joined with the superior mesenteric vein. To our knowledge, in the literature, there is just only one case of a true ectopic, locally vascularized spleen in the pelvis.

  6. Local fat treatments: classification proposal

    PubMed Central

    Pinto, Hernán

    2016-01-01

    ABSTRACT The poor understanding of the real, intimate action mechanisms behind any aesthetic procedures is a huge problem for many Aesthetic physicians. In addition, nomenclature of and regarding any procedure has become a true barrier when speaking about medical knowledge in the Aesthetic Medicine field since marketing and science often collide one another. Medical procedures for localized fat reduction are very different from each other and it is, at least, inaccurate to refer to all of them plainly as “fat reduction methods.” A specific classification has become urgent and its categories should be able to imply what each method entails. For this classification proposal, “reversibility,” “membrane disruption or inflammation,” and “action selectivity,” have been the selected criteria. PMID:27144093

  7. Observer Localization in Multiverse Theories

    NASA Astrophysics Data System (ADS)

    Hutter, Marcus

    2011-11-01

    The progression of theories suggested for our world, from ego- to geo- to helio-centric models to universe and multiverse theories and beyond, shows one tendency: The size of the described worlds increases, with humans being expelled from their center to ever more remote and random locations. If pushed too far, a potential theory of everything (TOE) is actually more a theories of nothing (TON). Indeed such theories have already been developed. I show that including observer localization into such theories is necessary and su_cient to avoid this problem. I develop a quantitative recipe to identify TOEs and distinguish them from TONs and theories in-between. This precisely shows what the problem is with some recently suggested universal TOEs.

  8. Relative Locality in Curved Spacetime

    NASA Astrophysics Data System (ADS)

    Kowalski-Glikman, Jerzy; Rosati, Giacomo

    2013-07-01

    In this paper we construct the action describing dynamics of the particle moving in curved spacetime, with a nontrivial momentum space geometry. Curved momentum space is the core feature of theories where relative locality effects are present. So far aspects of nonlinearities in momentum space have been studied only for flat or constantly expanding (de Sitter) spacetimes, relying on their maximally symmetric nature. The extension of curved momentum space frameworks to arbitrary spacetime geometries could be relevant for the opportunities to test Planck-scale curvature/deformation of particles momentum space. As a first example of this construction we describe the particle with κ-Poincaré momentum space on a circular orbit in Schwarzschild spacetime, where the contributes of momentum space curvature turn out to be negligible. The analysis of this problem relies crucially on the solution of the soccer ball problem.

  9. Biomarkers in localized prostate cancer.

    PubMed

    Ferro, Matteo; Buonerba, Carlo; Terracciano, Daniela; Lucarelli, Giuseppe; Cosimato, Vincenzo; Bottero, Danilo; Deliu, Victor M; Ditonno, Pasquale; Perdonà, Sisto; Autorino, Riccardo; Coman, Ioman; De Placido, Sabino; Di Lorenzo, Giuseppe; De Cobelli, Ottavio

    2016-02-01

    Biomarkers can improve prostate cancer diagnosis and treatment. Accuracy of prostate-specific antigen (PSA) for early diagnosis of prostate cancer is not satisfactory, as it is an organ- but not cancer-specific biomarker, and it can be improved by using models that incorporate PSA along with other test results, such as prostate cancer antigen 3, the molecular forms of PSA (proPSA, benign PSA and intact PSA), as well as kallikreins. Recent reports suggest that new tools may be provided by metabolomic studies as shown by preliminary data on sarcosine. Additional molecular biomarkers have been identified by the use of genomics, proteomics and metabolomics. We review the most relevant biomarkers for early diagnosis and management of localized prostate cancer.

  10. Anticipation of Local Epidemics Management

    NASA Astrophysics Data System (ADS)

    Krivy, Ivan

    2008-10-01

    Local epidemics are systems where temporary elements (infected ones) are distributed over a hexagonal or rectangular network of permanent elements (humans). Their relation of being neighbors is considered to be constant. The systems of this type can occur in rest homes, institutes of social care, nursery schools, summer camps, prisons, etc. The process of the infection propagation depends on the neighborhood relation and on the actual distance of the neighbors. It is influenced by delays during the transitive affecting the people and by random effects, the size of which can be diminished by the intervention of health care service. The anticipation of the health care intervention effects was supported by simulation models implemented by means of the object-oriented programming.

  11. Local geometry of isoscalar surfaces.

    PubMed

    Dopazo, César; Martín, Jesús; Hierro, Juan

    2007-11-01

    An inert dynamically passive scalar in a constant density fluid forced by a statistically homogeneous field of turbulence has been investigated using the results of a 256(3) grid direct numerical simulation. Mixing characteristics are characterized in terms of either principal curvatures or mean and Gauss curvatures. The most probable small-scale scalar geometries are flat and tilelike isosurfaces. Preliminary correlations between flow and scalar small-scale structures associate highly curved saddle points with large-strain regions and elliptic points with vorticity-dominated zones. The concavity of the scalar profiles along the isosurface normal coordinate xn correlates well with negative mean curvatures, Gauss curvatures displaying any sign, which correspond to scalar minima, tiles, or saddle points; on the other hand, convexity along xn is associated with positive mean curvatures, Gauss curvatures ranging from negative to positive signs, featuring maxima, tiles, or saddle points; inflection points along xn correlate well with small values of the mean curvature and zero or negative values of kg, corresponding to plane isosurfaces or saddle points with curvatures of equal and opposite signs. Small values of the scalar gradient are associated with elliptic points, either concave or convex (kg>0) , for both concave and convex scalar profiles along xn. Large values of the scalar gradient (or, equivalently, scalar fluctuation dissipation rates) are generally connected with small values of the Gauss curvature (either flat or moderate-curvature tilelike local geometries), with both concave and convex scalar profiles along xn equally probable. Vortical local flow structures correlate well with small and moderate values of the scalar gradient, while strain-dominated regions are associated with large values.

  12. Disease Localization in Multilayer Networks

    NASA Astrophysics Data System (ADS)

    de Arruda, Guilherme Ferraz; Cozzo, Emanuele; Peixoto, Tiago P.; Rodrigues, Francisco A.; Moreno, Yamir

    2017-01-01

    We present a continuous formulation of epidemic spreading on multilayer networks using a tensorial representation, extending the models of monoplex networks to this context. We derive analytical expressions for the epidemic threshold of the susceptible-infected-susceptible (SIS) and susceptible-infected-recovered dynamics, as well as upper and lower bounds for the disease prevalence in the steady state for the SIS scenario. Using the quasistationary state method, we numerically show the existence of disease localization and the emergence of two or more susceptibility peaks, which are characterized analytically and numerically through the inverse participation ratio. At variance with what is observed in single-layer networks, we show that disease localization takes place on the layers and not on the nodes of a given layer. Furthermore, when mapping the critical dynamics to an eigenvalue problem, we observe a characteristic transition in the eigenvalue spectra of the supra-contact tensor as a function of the ratio of two spreading rates: If the rate at which the disease spreads within a layer is comparable to the spreading rate across layers, the individual spectra of each layer merge with the coupling between layers. Finally, we report on an interesting phenomenon, the barrier effect; i.e., for a three-layer configuration, when the layer with the lowest eigenvalue is located at the center of the line, it can effectively act as a barrier to the disease. The formalism introduced here provides a unifying mathematical approach to disease contagion in multiplex systems, opening new possibilities for the study of spreading processes.

  13. The Local Volume Legacy Survey

    NASA Astrophysics Data System (ADS)

    Lee, J. C.; Kennicutt, R. C.; Engelbracht, C. W.; Calzetti, D.; Dale, D. A.; Gordon, K. D.; Dalcanton, J. J.; Skillman, E.; Begum, A.; Funes, J. G.; Gil de Paz, A.; Johnson, B.; Sakai, S.; van Zee, L.; Walter, F.; Weisz, D.; Williams, B.; Wu, Y.; Block, M.

    2008-10-01

    We introduce the Local Volume Legacy (LVL), a Spitzer Cycle 4 IRAC and MIPS Legacy survey of a complete sample of 258 galaxies within 11 Mpc. The broad goal of LVL is to provide critical insight into two of the primary processes that shape the growth of galaxies: star formation and its interaction with the interstellar medium. This goal will be achieved by investigating the spatially-resolved star formation, dust, and red stellar populations of galaxies that have been drawn from a statistically robust local sample, in which the full diversity of galaxy properties (e.g., luminosities, surface brightnesses, metallicities) are represented. Our tiered sample includes: (1) all known galaxies inside a sub-volume bounded by 3.5 Mpc, and (2) an unbiased sample of S-Irr galaxies within the larger, and more representative, 11 Mpc sphere. LVL will produce a multi-wavelength census of the Galactic neighborhood, extending to the faintest limits of the galactic luminosity function and exploiting the highest spatial resolution and absolute depth achievable with Spitzer. Our rich suite of ancillary data includes Hα and UV imaging from the GALEX 11HUGS and NGS surveys, stellar population mapping from the HST ANGST Treasury survey, H I mapping with the VLA and GMRT, optical broad-band imaging and spectroscopy. LVL will homogeneously fill in critical gaps in the current Spitzer coverage of the LV, provide SED coverage from the ultraviolet to the far-infrared, and thus supply the community with a core archival dataset on the Galactic neighborhood.

  14. Localization of quantum Bernoulli noises

    SciTech Connect

    Wang, Caishi; Zhang, Jihong

    2013-10-15

    The family (∂{sub k},∂{sub k}{sup *}){sub k≥0} of annihilation and creation operators acting on square integrable functionals of a Bernoulli process Z= (Z{sub k}){sub k⩾0} can be interpreted as quantum Bernoulli noises. In this note we consider the operator family (ℓ{sub k},ℓ{sub k}{sup *}){sub k≥0}, where ℓ{sub k}=∂{sub k}E{sub k} with E{sub k} being the conditional expectation (operator) given σ-field σ(Z{sub j}; 0 ⩽j⩽k). We show that ℓ{sub k} (resp. ℓ{sub k}{sup *}) is essentially a kind of localization of the annihilation operator ∂{sub k} (resp. creation operator ∂{sub k}{sup *}). We examine properties of the family (ℓ{sub k},ℓ{sub k}{sup *}){sub k≥0} and prove, among other things, that ℓ{sub k} and ℓ{sub k}{sup *} satisfy a local canonical anti-communication relation and (ℓ{sub k}{sup *}){sub k≥0} forms a mutually orthogonal operator sequence although each ℓ{sub k} is not a projection operator. We find that the operator series Σ{sub k=0}{sup ∞}ℓ{sub k}{sup *}Xℓ{sub k} converges in the strong operator topology for each bounded operator X acting on square integrable functionals of Z. In particular we get an explicit sum of the operator series Σ{sub k=0}{sup ∞}ℓ{sub k}{sup *}ℓ{sub k}. A useful norm estimate on Σ{sub k=0}{sup ∞}ℓ{sub k}{sup *}Xℓ{sub k} is also obtained. Finally we show applications of our main results to quantum dynamical semigroups and quantum probability.

  15. Local critical power is an index of local endurance.

    PubMed

    Le Chevalier, J M; Vandewalle, H; Thépaut-Mathieu, C; Stein, J F; Caplan, L

    2000-01-01

    The hypothesis that critical power (CP) is significantly lower than the maximal aerobic power of the knee extensors has been tested in nine endurance-trained subjects, seven gymnasts and seven weight lifters. CP was calculated as being equal to the slope of the linear relationship between exhaustion time and work performed at exhaustion on a knee-extension ergometer. CP was compared with the power output at the end of a progressive knee-extension exercise (P(peak)) and the power outputs corresponding to exhaustion times equal to 4 (P(4 min)), 6 (P(6 min)), 8 (P(8 min)) and 10 min (P(10 min)), calculated according to the linear relationship between work and exhaustion time. The hypothesis that CP corresponds to a steady state in metabolic and physiological parameters was tested in the gymnasts and the weight lifters by comparing CP with the fatigue thresholds of the integrated electromyogram (iEMG(FT)), lactate level (La(FT)), oxygen uptake (VO(2FT)) and heart rate (HR(FT)). The results of the present study demonstrate that the value of CP of a local exercise cannot be considered as the equivalent of the maximal aerobic power for general exercises. The values of P(4 min), P(6 min), P(8 min), P(10 min) and P(peak) were significantly higher than CP, and corresponded to 138, 126, 119, 115 and 151% CP, respectively. The results of the present study indicate that CP can be considered as an index of muscular endurance. Indeed, La(FT), iEMG (FT), VO(2FT) and HR(FT) were not significantly different from CP. All of these fatigue thresholds were significantly correlated with CP (r > 0.92). Moreover, the highest coefficient of correlation (r=0.71; P < 0.01) between the percentage of maximal aerobic power in cycling that corresponds to a blood lactate concentration of 4 mmol x l(-1) (OBLA%) and the different local aerobic indices was observed with CP.

  16. The Local Group: Our Galactic Neighborhood.

    ERIC Educational Resources Information Center

    Hodge, Paul

    1987-01-01

    Presents information on the properties and largest spirals of the Local Group galaxies. Explains the three categories of galaxies, identifies the brightest members of the Local Group, and discusses recent discoveries within the group. (ML)

  17. Method for localizing heating in tumor tissue

    DOEpatents

    Doss, James D.; McCabe, Charles W.

    1977-04-12

    A method for a localized tissue heating of tumors is disclosed. Localized radio frequency current fields are produced with specific electrode configurations. Several electrode configurations are disclosed, enabling variations in electrical and thermal properties of tissues to be exploited.

  18. Climate adaptation: Cultural knowledge and local risks

    NASA Astrophysics Data System (ADS)

    Strauss, Sarah

    2015-07-01

    A focus on African American communities on the Eastern Shore of Maryland highlights the ways that local cultural knowledge differs from place to place, developing understanding of local climate risks and resources for adaptation.

  19. Local Employment Initiatives: Some Recent Developments.

    ERIC Educational Resources Information Center

    Kuenstler, Peter

    1988-01-01

    Intergovernmental and governmental measures to promote local employment initiatives are underway in Europe. Local authorities are assuming responsibility for economic development in their areas, and the private sector is providing direct or indirect support. (SK)

  20. Local anesthetics for facial plastic procedures.

    PubMed

    Ahlstrom, Karen K; Frodel, John L

    2002-02-01

    The use of local anesthetic in facial plastic surgical procedures is well established as an effective and safe mode of anesthesia delivery. Local infiltration of anesthesia may be used alone for minor surgical procedures, or it may be used with general anesthesia or intravenous sedation and analgesia for more complex, lengthy procedures. When considered independently, the use of local anesthetic agents has undeniable limitations. Local anesthetics can cause toxicity and side effects. Injection of local anesthetics for subcutaneous infiltration frequently is painful until sensory anesthesia occurs. Local anesthetics have limited efficacy with respect to the intensity and duration of sensory blockade that can be achieved. In some situations, use of local anesthesia with the maintenance of an awake patient also may be undesirable for the surgeon and impractical for the patient. Despite these shortcomings, local anesthetics are fundamentally ideal for use in facial plastic surgery.

  1. Local Authority Education in a Democratic Scotland.

    ERIC Educational Resources Information Center

    Fairley, John

    1998-01-01

    Establishment of a Scottish Parliament in 1999 will pose questions about its relationship to local authorities and their role in education. Possible scenarios are discussed in which Parliament pursues centralization or decentralization in education policy. Recent reforms to local government structures may allow local councils to become partners of…

  2. Local Observed-Score Kernel Equating

    ERIC Educational Resources Information Center

    Wiberg, Marie; van der Linden, Wim J.; von Davier, Alina A.

    2014-01-01

    Three local observed-score kernel equating methods that integrate methods from the local equating and kernel equating frameworks are proposed. The new methods were compared with their earlier counterparts with respect to such measures as bias--as defined by Lord's criterion of equity--and percent relative error. The local kernel item response…

  3. Choices of local anesthetics for ocular surgery.

    PubMed

    Cass, Gary D

    2006-06-01

    This article will help clinicians to be aware of their choices of local anesthetic solutions before they make their ocular anesthetic plan based on the specific requirements of the patient, the surgical procedure, and the properties of the local anesthetic. Choices of local anesthetic solutions and additives for both topical anesthesia and conduction blockade are discussed.

  4. Using Local Materials To Teach Writing.

    ERIC Educational Resources Information Center

    Ness, Verna M.

    1997-01-01

    English teachers in foreign countries frequently encounter a lack of materials in English dealing with local topics of interest to students. One suggestion is to use materials written in English for tourists, such as travel brochures or local maps. These materials are readily available in most places at little or no cost from local travel agencies…

  5. 34 CFR 361.15 - Local administration.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 34 Education 2 2012-07-01 2012-07-01 false Local administration. 361.15 Section 361.15 Education... Other Requirements for Vocational Rehabilitation Services Administration § 361.15 Local administration. (a) If the State plan provides for the administration of the plan by a local agency, the...

  6. 34 CFR 361.15 - Local administration.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 34 Education 2 2014-07-01 2013-07-01 true Local administration. 361.15 Section 361.15 Education... Other Requirements for Vocational Rehabilitation Services Administration § 361.15 Local administration. (a) If the State plan provides for the administration of the plan by a local agency, the...

  7. 34 CFR 361.15 - Local administration.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 34 Education 2 2013-07-01 2013-07-01 false Local administration. 361.15 Section 361.15 Education... Other Requirements for Vocational Rehabilitation Services Administration § 361.15 Local administration. (a) If the State plan provides for the administration of the plan by a local agency, the...

  8. 34 CFR 361.15 - Local administration.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 34 Education 2 2011-07-01 2010-07-01 true Local administration. 361.15 Section 361.15 Education... Other Requirements for Vocational Rehabilitation Services Administration § 361.15 Local administration. (a) If the State plan provides for the administration of the plan by a local agency, the...

  9. 34 CFR 361.15 - Local administration.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Local administration. 361.15 Section 361.15 Education... Other Requirements for Vocational Rehabilitation Services Administration § 361.15 Local administration. (a) If the State plan provides for the administration of the plan by a local agency, the...

  10. Local Service Learning in Teacher Preparation Program

    ERIC Educational Resources Information Center

    Nuangchalerm, Prasart

    2016-01-01

    The local knowledge is simply integrated in education and learning process. This study aims to promote local knowledge in school through service learning. The learning process is employed herbal plants to reinforce students learn how to sustain local knowledge with modern life and 21st century classroom. Participants consisted of 42 pre-service…

  11. 2 CFR 200.64 - Local government.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 2 Grants and Agreements 1 2014-01-01 2014-01-01 false Local government. 200.64 Section 200.64... REQUIREMENTS FOR FEDERAL AWARDS Acronyms and Definitions Acronyms § 200.64 Local government. Local government means any unit of government within a state, including a: (a) County; (b) Borough; (c) Municipality;...

  12. 33 CFR 238.9 - Local cooperation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Local cooperation. 238.9 Section 238.9 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF... Local cooperation. (a) Cost sharing and other provisions of local cooperation shall be in...

  13. Diverse Local Literacies and Standardizing Policies

    ERIC Educational Resources Information Center

    Bomer, Randy; Maloch, Beth

    2012-01-01

    The authors challenge educators to speculate on how to address local issues in a world that is increasingly diverse and global. This Research and Policy section emphasizes the seemingly paradoxical importance of both local and global literacy, illuminating the tension between the preservation of local literacy and a global standardization of…

  14. 10 CFR 440.13 - Local applications.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 3 2010-01-01 2010-01-01 false Local applications. 440.13 Section 440.13 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION WEATHERIZATION ASSISTANCE FOR LOW-INCOME PERSONS § 440.13 Local applications. (a) The Support Office Director shall give written notice to all local applicants throughout...

  15. Localism: The Changing Picture for Adult Learning

    ERIC Educational Resources Information Center

    Lamb, Penny

    2012-01-01

    The rapidly changing picture on localism and the government's focus on local economic growth have significant implications for adult learning and skills providers in England. Government now sees a sense of place as key to economic growth and recognises the need for a renewed debate on how business and state interact with localities. There is a…

  16. Colleges Chew on Local-Food Phenomenon

    ERIC Educational Resources Information Center

    Carlson, Scott

    2008-01-01

    Nationally, students have pushed hard for local-food programs, and the amount of local produce in the dining hall is sometimes used as a litmus test for a college's overall commitment to sustainability. Local food has been a hot topic in popular culture in recent years, thanks in part to books by Michael Pollan and Barbara Kingsolver, movies like…

  17. 32 CFR 1602.14 - Local board.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Local board. 1602.14 Section 1602.14 National Defense Other Regulations Relating to National Defense SELECTIVE SERVICE SYSTEM DEFINITIONS § 1602.14 Local board. A local board or a panel thereof of the Selective Service System is a group of not...

  18. 33 CFR 238.9 - Local cooperation.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false Local cooperation. 238.9 Section 238.9 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF... Local cooperation. (a) Cost sharing and other provisions of local cooperation shall be in...

  19. 33 CFR 238.9 - Local cooperation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false Local cooperation. 238.9 Section 238.9 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF... Local cooperation. (a) Cost sharing and other provisions of local cooperation shall be in...

  20. 33 CFR 238.9 - Local cooperation.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Local cooperation. 238.9 Section 238.9 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF... Local cooperation. (a) Cost sharing and other provisions of local cooperation shall be in...

  1. 33 CFR 238.9 - Local cooperation.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Local cooperation. 238.9 Section 238.9 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF... Local cooperation. (a) Cost sharing and other provisions of local cooperation shall be in...

  2. State and Local Compliance: a National Report.

    ERIC Educational Resources Information Center

    Beuke, Vernon

    1981-01-01

    Discusses the Abt Study of State and Local Compliance which sought to describe state and local implementation of the provisions of the Vocational Education Amendments of 1965; to understand ways in which local environment affects compliance; and to provide Congress with recommendations for achieving greater adherence to federal intent. (JOW)

  3. The local dark matter density

    NASA Astrophysics Data System (ADS)

    Read, J. I.

    2014-06-01

    I review current efforts to measure the mean density of dark matter near the Sun. This encodes valuable dynamical information about our Galaxy and is also of great importance for ‘direct detection’ dark matter experiments. I discuss theoretical expectations in our current cosmology; the theory behind mass modelling of the Galaxy; and I show how combining local and global measures probes the shape of the Milky Way dark matter halo and the possible presence of a ‘dark disc’. I stress the strengths and weaknesses of different methodologies and highlight the continuing need for detailed tests on mock data—particularly in the light of recently discovered evidence for disequilibria in the Milky Way disc. I collate the latest measurements of ρdm and show that, once the baryonic surface density contribution Σb is normalized across different groups, there is remarkably good agreement. Compiling data from the literature, I estimate Σb = 54.2 ± 4.9 M⊙pc-2, where the dominant source of uncertainty is in the H i gas contribution. Assuming this contribution from the baryons, I highlight several recent measurements of ρdm in order of increasing data complexity and prior, and, correspondingly, decreasing formal error bars. Comparing these measurements with spherical extrapolations from the Milky Way’s rotation curve, I show that the Milky Way is consistent with having a spherical dark matter halo at R0 ˜ 8 kpc. The very latest measures of ρdm based on ˜10 000 stars from the Sloan Digital Sky Survey appear to favour little halo flattening at R0, suggesting that the Galaxy has a rather weak dark matter disc, with a correspondingly quiescent merger history. I caution, however, that this result hinges on there being no large systematics that remain to be uncovered in the SDSS data, and on the local baryonic surface density being Σb ˜ 55 M⊙pc-2. I conclude by discussing how the new Gaia satellite will be transformative. We will obtain much tighter

  4. Completing HST's Local Volume Legacy

    NASA Astrophysics Data System (ADS)

    Dalcanton, Julianne

    2007-07-01

    Nearby galaxies offer one of the few laboratories within which stellar populations can be tied to multi-wavelength observations. They are thus essential for calibrating and interpreting key astrophysical observables, such as broad-band luminosities, durations and energy input from starbursts, and timescales of UV, H-alpha, and FIR emission. The study of stellar populations in nearby galaxies requires high-resolution observations with HST, but HST's legacy for this limited set of galaxies remains incomplete.As a first attempt to establish this legacy, The ACS Nearby Galaxy Survey Treasury {ANGST} began observations in late 2006. ANGST was designed to carry out a uniform multi-color survey of a volume-limited sample of 70 nearby galaxies that could be used for systematic studies of resolved stellar populations. The resulting data provide nuanced constraints on the processes which govern star formation and galaxy evolution, for a well-defined population of galaxies. All photometry for the survey has been publicly released.However, the failure of ACS 4.5 months after ANGST began taking data led to a drastic reduction in the planned survey. The loss is two-fold. First, the goals of completeness and uniformity were greatly compromised, impacting global comparison studies. Second, the variety of observed star formation histories was reduced. Given that we have never found two galaxies with identical star formation histories, and fully sampling the population allows us to catch those few systems whose star formation rates and metallicities place the strongest constraints on key astrophysical processes.Here we propose WFPC2 observations of all remaining galaxies within the Local Volume {D<3.5Mpc} for which current HST observations are insufficient for meaningful stellar population studies. We will use these observations for research on the star formation histories of individual galaxies and the Local Volume, detailed calibrations of star formation rate indicators, and the

  5. Localization of weakly disordered flat band states

    NASA Astrophysics Data System (ADS)

    Leykam, Daniel; Bodyfelt, Joshua D.; Desyatnikov, Anton S.; Flach, Sergej

    2017-01-01

    Certain tight binding lattices host macroscopically degenerate flat spectral bands. Their origin is rooted in local symmetries of the lattice, with destructive interference leading to the existence of compact localized eigenstates. We study the robustness of this localization to disorder in different classes of flat band lattices in one and two dimensions. Depending on the flat band class, the flat band states can either be robust, preserving their strong localization for weak disorder W, or they are destroyed and acquire large localization lengths ξ that diverge with a variety of unconventional exponents ν, ξ 1 / W ν .

  6. Local anaesthesia outside the operating room.

    PubMed

    Chan, S K; Karmakar, M K; Chui, P T

    2002-04-01

    An increasing number of minor surgical procedures are performed under local anaesthesia in clinical settings outside the operating room, where monitoring and resuscitation equipment--as well as personnel skilled in resuscitation--may not be readily available. Serious adverse effects and even fatalities may result from the use of local anaesthetic agents, arising from a variety of causes such as systemic toxicity, allergy, vasovagal syncope, and reaction to additives present in the local anaesthetic. This article briefly reviews the pharmacology of local anaesthetic agents, and describes various techniques commonly used for local anaesthesia, with special emphasis on safety. Clinical features of toxicity, and its differential diagnosis and management, are also discussed.

  7. Coordinating locally 'owned' treatment guidelines.

    PubMed

    Collier, J; Picton, C; Littlejohns, P

    1994-01-01

    South West Thames Regional Health Authority established and commissioned a regional guidelines unit to coordinate the introduction of a set of treatment guidelines on the management of common medical emergencies into all the acute intaking National Health Service (NHS) hospitals throughout the region. All hospitals were offered a set of template guidelines to be used at their discretion for producing their own customised equivalent. They were also offered full typing and production facilities, together with printing costs if publication was achieved by a target deadline (1 August 1993). In 11 of the 14 NHS hospitals guidelines were available to hospital staff by the target deadline, and one set was produced for a non-NHS hospital. In two hospitals the target date was not met, and one other declined to take part. As part of the project the unit assessed the extent to which the published guidelines were adapted to meet the requirements of each individual hospital. The template offered guidelines on 34 topic titles. No hospital used all core titles of the original template; titles were omitted or replaced in some, and added in others. Where the original guideline titles were used, there was almost always some customisation--changes in sentence structure, names or contact numbers, alterations in drugs and doses or the addition or omission of entire sections. By using an established resource, sets of customised, locally determined treatment guidelines were introduced with relative ease into most of the acute hospitals in a UK health region.

  8. Segmentation in local hospital markets.

    PubMed

    Dranove, D; White, W D; Wu, L

    1993-01-01

    This study examines evidence of market segmentation on the basis of patients' insurance status, demographic characteristics, and medical condition in selected local markets in California in the years 1983 and 1989. Substantial differences exist in the probability patients may be admitted to particular hospitals based on insurance coverage, particularly Medicaid, and race. Segmentation based on insurance and race is related to hospital characteristics, but not the characteristics of the hospital's community. Medicaid patients are more likely to go to hospitals with lower costs and fewer service offerings. Privately insured patients go to hospitals offering more services, although cost concerns are increasing. Hispanic patients also go to low-cost hospitals, ceteris paribus. Results indicate little evidence of segmentation based on medical condition in either 1983 or 1989, suggesting that "centers of excellence" have yet to play an important role in patient choice of hospital. The authors found that distance matters, and that patients prefer nearby hospitals, moreso for some medical conditions than others, in ways consistent with economic theories of consumer choice.

  9. Multimodality localization of epileptic foci

    NASA Astrophysics Data System (ADS)

    Desco, Manuel; Pascau, Javier; Pozo, M. A.; Santos, Andres; Reig, Santiago; Gispert, Juan D.; Garcia-Barreno, Pedro

    2001-05-01

    This paper presents a multimodality approach for the localization of epileptic foci using PET, MRI and EEG combined without the need of external markers. Mutual Information algorithm is used for MRI-PET registration. Dipole coordinates (provided by BESA software) are projected onto the MRI using a specifically developed algorithm. The four anatomical references used for electrode positioning (nasion, inion and two preauricular points) are located on the MRI using a triplanar viewer combined with a surface-rendering tool. Geometric transformation using deformation of the ideal sphere used for dipole calculations is then applied to match the patient's brain size and shape. Eight treatment-refractory epileptic patients have been studied. The combination of the anatomical information from the MRI, hipoperfusion areas in PET and dipole position and orientation helped the physician in the diagnosis of epileptic focus location. Neurosurgery was not indicated for patients where PET and dipole results were inconsistent; in two cases it was clinically indicated despite the mismatch, showing a negative follow up. The multimodality approach presented does not require external markers for dipole projection onto the MRI, this being the main difference with previous methods. The proposed method may play an important role in the indication of surgery for treatment- refractory epileptic patients.

  10. Carrier localization in gallium nitride

    SciTech Connect

    Wetzel, C.; Walukiewicz, W.; Haller, E.E.

    1996-09-01

    In wide bandgap GaN, a large number of interesting and important scientific questions remain to be answered. For example, the large free electron concentration reaching 10{sup 19} to 10{sup 20} cm{sup - 3} in nominally undoped material are ascribed to intrinsic defects because no chemical impurity has been found at such high concentrations. According to theoretical models, a nitrogen vacancy acts as a donor but its formation energy is very large in n-type materials, making this suggestion controversial. We have investigated the nature of this yet unidentified donor at large hydrostatic pressure. Results from infrared reflection and Raman scattering indicate strong evidence for localization of free carriers by large pressures. The carrier density is drastically decreased by two orders of magnitude between 20 and 30 GPa. Several techniques provide independent evidence for results in earlier reports and present the first quantitative analysis. A possible interpretation of this effect in terms of the resonant donor level is presented.

  11. Matryoshka locally resonant sonic crystal.

    PubMed

    Elford, Daniel P; Chalmers, Luke; Kusmartsev, Feodor V; Swallowe, Gerry M

    2011-11-01

    The results of numerical modeling of sonic crystals with resonant array elements are reported. The investigated resonant elements include plain slotted cylinders as well as their various combinations, in particular, Russian doll or Matryoshka configurations. The acoustic band structure and transmission characteristics of such systems have been computed with the use of finite element methods. The general concept of a locally resonant sonic crystal is proposed that utilizes acoustic resonances to form additional band gaps that are decoupled from Bragg gaps. An existence of a separate attenuation mechanism associated with the resonant elements that increases performance in the lower frequency regime has been identified. The results show a formation of broad band gaps positioned significantly below the first Bragg frequency. For low frequency broadband attenuation, a most optimal configuration is the Matryoshka sonic crystal, where each scattering unit is composed of multiple concentric slotted cylinders. This system forms numerous gaps in the lower frequency regime, below Bragg bands, while maintaining a reduced crystal size viable for noise barrier technology. The finding opens alternative perspectives for the construction of sound barriers in the low frequency range usually inaccessible by traditional means including conventional sonic crystals.

  12. Matryoshka locally resonant sonic crystal

    NASA Astrophysics Data System (ADS)

    Elford, Daniel P.; Chalmers, Luke; Kusmartsev, Feodor V.; Swallowe, Gerry M.

    The results of numerical modelling of sonic crystals with resonant array elements are reported. The investigated resonant elements include plain slotted cylinders as well as various their combinations, in particular, Russian doll or Matryoshka configurations. The acoustic band structure and transmission characteristics of such systems have been computed with the use of finite element methods. The general concept of a locally resonant sonic crystal is proposed, which utilises acoustic resonances to form additional band gaps that are decoupled from Bragg gaps. An existence of a separate attenuation mechanism associated with the resonant elements, which increases performance in the lower frequency regime has been identified. The results show a formation of broad band gaps positioned significantly below the first Bragg frequency. For low frequency broadband attenuation a most optimal configuration is the Matryoshka sonic crystal, where each scattering unit is composed of multiple concentric slotted cylinders. This system forms numerous gaps in the lower frequency regime, below Bragg bands, whilst maintaining a reduced crystal size viable for noise barrier technology. The finding opens new perspectives for construction of sound barriers in the low frequency range usually inaccessible by traditional means including conventional sonic crystals.

  13. Intrinsic Localized Modes in Proteins

    PubMed Central

    Nicolaï, Adrien; Delarue, Patrice; Senet, Patrick

    2015-01-01

    Protein dynamics is essential for proteins to function. Here we predicted the existence of rare, large nonlinear excitations, termed intrinsic localized modes (ILMs), of the main chain of proteins based on all-atom molecular dynamics simulations of two fast-folder proteins and of a rigid α/β protein at 300 K and at 380 K in solution. These nonlinear excitations arise from the anharmonicity of the protein dynamics. The ILMs were detected by computing the Shannon entropy of the protein main-chain fluctuations. In the non-native state (significantly explored at 380 K), the probability of their excitation was increased by a factor between 9 and 28 for the fast-folder proteins and by a factor 2 for the rigid protein. This enhancement in the non-native state was due to glycine, as demonstrated by simulations in which glycine was mutated to alanine. These ILMs might play a functional role in the flexible regions of proteins and in proteins in a non-native state (i.e. misfolded or unfolded states). PMID:26658321

  14. Hidden local symmetry and beyond

    NASA Astrophysics Data System (ADS)

    Yamawaki, Koichi

    Gerry Brown was a godfather of our hidden local symmetry (HLS) for the vector meson from the birth of the theory throughout his life. The HLS is originated from very nature of the nonlinear realization of the symmetry G based on the manifold G/H, and thus is universal to any physics based on the nonlinear realization. Here, I focus on the Higgs Lagrangian of the Standard Model (SM), which is shown to be equivalent to the nonlinear sigma model based on G/H = SU(2)L ×SU(2)R/SU(2)V with additional symmetry, the nonlinearly-realized scale symmetry. Then, the SM does have a dynamical gauge boson of the SU(2)V HLS, “SM ρ meson”, in addition to the Higgs as a pseudo-dilaton as well as the NG bosons to be absorbed in to the W and Z. Based on the recent work done with Matsuzaki and Ohki, I discuss a novel possibility that the SM ρ meson acquires kinetic term by the SM dynamics itself, which then stabilizes the skyrmion dormant in the SM as a viable candidate for the dark matter, what we call “dark SM skyrmion (DSMS)”.

  15. Improved Measure of Local Chirality

    SciTech Connect

    Terrence Draper; Andrei Alexandru; Ying Chen; Shao-Jing Dong; Ivan Horvath; Frank Lee; Nilmani Mathur; Harry B. Thacker; Sonali Tamhankar; Jianbo Zhang

    2004-06-01

    It is popular to probe the structure of the QCD vacuum indirectly by studying individual fermion eigenmodes, because this provides a natural way to filter out UV fluctuations. The double-peaking in the distribution of the local chiral orientation parameter (X) has been offered as evidence, by some, in support of a particular model of the vacuum. Here we caution that the X-distribution peaking varies significantly with various versions of the definition of X. Furthermore, each distribution varies little from that resulting from a random reshuffling of the left-handed (and independently the right-handed) fields, which destroys any QCD-induced left-right correlation; that is, the double-peaking is mostly a phase-space effect. We propose a new universal definition of the X parameter whose distribution is uniform for randomly reshuffled fields. Any deviations from uniformity for actual data can then be directly attributable to QCD-induced dynamics. We find that the familiar double peak disappears.

  16. Enzyme function is regulated by its localization.

    PubMed

    Gifford, Stacey M; Meyer, Pablo

    2015-12-01

    To better understand how enzyme localization affects enzyme activity we studied the cellular localization of the glycosyltransferase MurG, an enzyme necessary for cell wall synthesis at the spore during sporulation in the bacterium Bacillus subtilis. During sporulation MurG was gradually enriched to the membrane at the forespore and point mutations in a MurG helical domain disrupting its localization to the membrane caused severe sporulation defects, but did not affect localization nor caused detectable defects during exponential growth. We found that this localization is dependent on the phospholipid cardiolipin, as in strains where the cardiolipin-synthesizing genes were deleted, MurG levels were diminished at the forespore. Furthermore, in this cardiolipin-less strain, MurG localization during sporulation was rescued by external addition of purified cardiolipin. These results support localization as a critical factor in the regulation of proper enzyme function and catalysis.

  17. Metastable localization of diseases in complex networks

    NASA Astrophysics Data System (ADS)

    Ferreira, R. S.; da Costa, R. A.; Dorogovtsev, S. N.; Mendes, J. F. F.

    2016-12-01

    We describe the phenomenon of localization in the epidemic susceptible-infective-susceptible model on highly heterogeneous networks in which strongly connected nodes (hubs) play the role of centers of localization. We find that in this model the localized states below the epidemic threshold are metastable. The longevity and scale of the metastable outbreaks do not show a sharp localization transition; instead there is a smooth crossover from localized to delocalized states as we approach the epidemic threshold from below. Analyzing these long-lasting local outbreaks for a random regular graph with a hub, we show how this localization can be detected from the shape of the distribution of the number of infective nodes.

  18. What's new in local anaesthesia?

    PubMed

    Malamed, Stanley

    2009-01-01

    In this paper I have explored four areas of current interest to pain control in dentistry. Articaine HCl, the most recent addition to the dental LA armamentarium, has become a favoured drug in many, if not most, countries in which it is available. Rapid onset and improved hard- and soft-tissue penetration enable articaine HCl to be administered with great success as a mandibular infiltration, precluding the need, in most situations, to employ it by inferior alveolar nerve block. The 'question' about an increased risk of paresthesia following articaine administration via IANB has been answered by careful evaluation of case reports. C-CLAD systems have enabled the administration of LA to become much more comfortable, especially in the palate, and with accessory techniques such as the periodontal ligament injection (PDL, ILI). Two highly successful techniques, the AMSA and P-ASA, have been developed as a result of C-CLAD systems. Phentolamine mesylate (OraVerse) allows for the reversal of residual soft-tissue anaesthesia, decreasing its duration by approximately 50%. Reversal enables patients to 'feel normal' more quickly after dental treatment and should decrease the risk of traumatic injury to soft tissues. Knowledge of the maximum dosages of LAs to be administered to all patients, but to younger, lighter-weight patients in particular, is essential to safety. The prevention of LA overdose is more gratifying than managing this fear-inducing medical emergency. When used properly, local anaesthetics represent the safest and most effective drugs in all of medicine for the prevention and management of pain.

  19. Local conservation laws and the structure of the many-body localized states.

    PubMed

    Serbyn, Maksym; Papić, Z; Abanin, Dmitry A

    2013-09-20

    We construct a complete set of local integrals of motion that characterize the many-body localized (MBL) phase. Our approach relies on the assumption that local perturbations act locally on the eigenstates in the MBL phase, which is supported by numerical simulations of the random-field XXZ spin chain. We describe the structure of the eigenstates in the MBL phase and discuss the implications of local conservation laws for its nonequilibrium quantum dynamics. We argue that the many-body localization can be used to protect coherence in the system by suppressing relaxation between eigenstates with different local integrals of motion.

  20. A local approach for focussed Bayesian fusion

    NASA Astrophysics Data System (ADS)

    Sander, Jennifer; Heizmann, Michael; Goussev, Igor; Beyerer, Jürgen

    2009-04-01

    Local Bayesian fusion approaches aim to reduce high storage and computational costs of Bayesian fusion which is separated from fixed modeling assumptions. Using the small world formalism, we argue why this proceeding is conform with Bayesian theory. Then, we concentrate on the realization of local Bayesian fusion by focussing the fusion process solely on local regions that are task relevant with a high probability. The resulting local models correspond then to restricted versions of the original one. In a previous publication, we used bounds for the probability of misleading evidence to show the validity of the pre-evaluation of task specific knowledge and prior information which we perform to build local models. In this paper, we prove the validity of this proceeding using information theoretic arguments. For additional efficiency, local Bayesian fusion can be realized in a distributed manner. Here, several local Bayesian fusion tasks are evaluated and unified after the actual fusion process. For the practical realization of distributed local Bayesian fusion, software agents are predestinated. There is a natural analogy between the resulting agent based architecture and criminal investigations in real life. We show how this analogy can be used to improve the efficiency of distributed local Bayesian fusion additionally. Using a landscape model, we present an experimental study of distributed local Bayesian fusion in the field of reconnaissance, which highlights its high potential.

  1. When Gravity Fails: Local Search Topology

    NASA Technical Reports Server (NTRS)

    Frank, Jeremy; Cheeseman, Peter; Stutz, John; Lau, Sonie (Technical Monitor)

    1997-01-01

    Local search algorithms for combinatorial search problems frequently encounter a sequence of states in which it is impossible to improve the value of the objective function; moves through these regions, called {\\em plateau moves), dominate the time spent in local search. We analyze and characterize {\\em plateaus) for three different classes of randomly generated Boolean Satisfiability problems. We identify several interesting features of plateaus that impact the performance of local search algorithms. We show that local minima tend to be small but occasionally may be very large. We also show that local minima can be escaped without unsatisfying a large number of clauses, but that systematically searching for an escape route may be computationally expensive if the local minimum is large. We show that plateaus with exits, called benches, tend to be much larger than minima, and that some benches have very few exit states which local search can use to escape. We show that the solutions (i.e. global minima) of randomly generated problem instances form clusters, which behave similarly to local minima. We revisit several enhancements of local search algorithms and explain their performance in light of our results. Finally we discuss strategies for creating the next generation of local search algorithms.

  2. Local Estimators for Spacecraft Formation Flying

    NASA Technical Reports Server (NTRS)

    Fathpour, Nanaz; Hadaegh, Fred Y.; Mesbahi, Mehran; Nabi, Marzieh

    2011-01-01

    A formation estimation architecture for formation flying builds upon the local information exchange among multiple local estimators. Spacecraft formation flying involves the coordination of states among multiple spacecraft through relative sensing, inter-spacecraft communication, and control. Most existing formation flying estimation algorithms can only be supported via highly centralized, all-to-all, static relative sensing. New algorithms are needed that are scalable, modular, and robust to variations in the topology and link characteristics of the formation exchange network. These distributed algorithms should rely on a local information-exchange network, relaxing the assumptions on existing algorithms. In this research, it was shown that only local observability is required to design a formation estimator and control law. The approach relies on breaking up the overall information-exchange network into sequence of local subnetworks, and invoking an agreement-type filter to reach consensus among local estimators within each local network. State estimates were obtained by a set of local measurements that were passed through a set of communicating Kalman filters to reach an overall state estimation for the formation. An optimization approach was also presented by means of which diffused estimates over the network can be incorporated in the local estimates obtained by each estimator via local measurements. This approach compares favorably with that obtained by a centralized Kalman filter, which requires complete knowledge of the raw measurement available to each estimator.

  3. On the concept of Bell’s local causality in local classical and quantum theory

    SciTech Connect

    Hofer-Szabó, Gábor; Vecsernyés, Péter

    2015-03-15

    The aim of this paper is to implement Bell’s notion of local causality into a framework, called local physical theory. This framework, based on the axioms of algebraic field theory, is broad enough to integrate both probabilistic and spatiotemporal concepts and also classical and quantum theories. Bell’s original idea of local causality will arise as the classical case of our definition. Classifying local physical theories by whether they obey local primitive causality, a property rendering the dynamics of the theory causal, we then investigate what is needed for a local physical theory to be locally causal. Finally, comparing local causality with the common cause principles and relating both to the Bell inequalities we find a nice parallelism: Bell inequalities cannot be derived neither from local causality nor from a common cause unless the local physical theory is classical or the common cause is commuting, respectively.

  4. On speciation of VOC localization

    NASA Astrophysics Data System (ADS)

    Chen, S.; Chang, J.; Wang, J.

    2011-12-01

    any shortcomings in VOC details and the simulation of oxidant levels remain unchanged. But for the detail emissions of VOC, one can find out the error with local VOC speciation and make corrections.

  5. Local tomographic phase microscopy from differential projections

    NASA Astrophysics Data System (ADS)

    Vishnyakov, G. N.; Levin, G. G.; Minaev, V. L.; Nekrasov, N. A.

    2016-12-01

    It is proposed to use local tomography for optical studies of the internal structure of transparent phase microscopic objects, for example, living cells. From among the many local tomography methods that exist, the algorithms of back projection summation (in which partial derivatives of projections are used as projection data) are chosen. The application of local tomography to living cells is reasonable because, using optical phase microscopy, one can easily obtain projection data in the form of first-order derivatives of projections applying the methods of differential interference contrast and shear interferometry. The mathematical fundamentals of local tomography in differential projections are considered, and a computer simulation of different local tomography methods is performed. A tomographic phase microscope and the results of reconstructing a local tomogram of an erythrocyte from a set of experimental differential projections are described.

  6. Expanding the principle of local distinguishability

    NASA Astrophysics Data System (ADS)

    Carmeli, Claudio; Heinosaari, Teiko; Schultz, Jussi; Toigo, Alessandro

    2015-04-01

    The principle of local distinguishability states that an arbitrary physical state of a bipartite system can be determined by the combined statistics of local measurements performed on the subsystems. A necessary and sufficient requirement for the local measurements is that each one must be able to distinguish between all pairs of states of the respective subsystems. We show that, if the task is changed into the determination of an arbitrary bipartite pure state, then at least in certain cases it is possible to restrict to local measurements which can distinguish all pure states but not all states. Moreover, we show that, if the local measurements are such that the purity of the bipartite state can be verified from the statistics without any prior assumption, then in these special cases also this property is carried over to the composite measurement. These surprising facts give evidence that the principle of local distinguishability may be expanded beyond its usual applicability.

  7. Localization in an Idealized Heterogeneous Elastic Sheet

    NASA Astrophysics Data System (ADS)

    Gurmessa, Bekele; Croll, Andrew B.

    2015-03-01

    Localized deformation is ubiquitous in many natural and engineering materials. Often times such deformations are associated to non-homogeneous strain fields in the materials. In this work we demonstrate the response of idealized non-homogenous elastic sheets to uniaxial compression. The idealized/patterned surface layers are created by selective ultraviolet/ozone (UVO) treatment of the top surface of polydimethylsiloxane (PDMS) using TEM grid mask. By controlling the exposure time of the UVO, samples ranging from continuous thin films to sets of isolated small plates were created. We show how local strains vary with location in a patterned sample, leading to a complex localization process Even at low strains. We also see that continuous regions form isotropic undulations upon compression which persist to high strains, well beyond where localization is observed in the patterned regions. Despite the complexity, the localized deformation profile can be adequately described with a simple elastic model when appropriate local boundary conditions are considered.

  8. Methods and strategies of object localization

    NASA Technical Reports Server (NTRS)

    Shao, Lejun; Volz, Richard A.

    1989-01-01

    An important property of an intelligent robot is to be able to determine the location of an object in 3-D space. A general object localization system structure is proposed, some important issues on localization discussed, and an overview given for current available object localization algorithms and systems. The algorithms reviewed are characterized by their feature extracting and matching strategies; the range finding methods; the types of locatable objects; and the mathematical formulating methods.

  9. Localizing the Fast Radio Burst 121102

    NASA Astrophysics Data System (ADS)

    Chatterjee, Shami; Wharton, Robert; Law, Casey J.; Hessels, Jason; Burke-Spolaor, Sarah; Bower, Geoffrey C.; Abruzzo, Matthew W.; Bassa, Cees; Butler, Bryan J.; Cordes, James M.; Paul, Demorest; Kaspi, Victoria M.; McLaughlin, Maura; Ransom, Scott M.; Scholz, Paul; Seymour, Andrew; Spitler, Laura; Tendulkar, Shriharsh P.; PALFA Survey Team; VLA+AO FRB121102 Simultaneous Campaign Team; EVN FRB121102 Campaign Team

    2017-01-01

    The precise localization of a fast radio burst and the identification of its host counterpart would allow constraints on their distances and energetics, and enable us to discriminate between various origin scenarios, from the local and mundane to the cosmological and exotic. Here we report on the results of an ongoing localization campaign on the repeating fast radio burst source, FRB 121102, with the VLA, Arecibo, and other telescopes.

  10. Local quanta, unitary inequivalence, and vacuum entanglement

    SciTech Connect

    Vázquez, Matías R. Rey, Marco del Westman, Hans León, Juan

    2014-12-15

    In this work we develop a formalism for describing localised quanta for a real-valued Klein–Gordon field in a one-dimensional box [0,R]. We quantise the field using non-stationary local modes which, at some arbitrarily chosen initial time, are completely localised within the left or the right side of the box. In this concrete set-up we directly face the problems inherent to a notion of local field excitations, usually thought of as elementary particles. Specifically, by computing the Bogoliubov coefficients relating local and standard (global) quantisations, we show that the local quantisation yields a Fock representation of the Canonical Commutation Relations (CCR) which is unitarily inequivalent to the standard one. In spite of this, we find that the local creators and annihilators remain well defined in the global Fock space F{sup G}, and so do the local number operators associated to the left and right partitions of the box. We end up with a useful mathematical toolbox to analyse and characterise local features of quantum states in F{sup G}. Specifically, an analysis of the global vacuum state |0{sub G}〉∈F{sup G} in terms of local number operators shows, as expected, the existence of entanglement between the left and right regions of the box. The local vacuum |0{sub L}〉∈F{sup L}, on the contrary, has a very different character. It is neither cyclic (with respect to any local algebra of operators) nor separating and displays no entanglement between left and right partitions. Further analysis shows that the global vacuum also exhibits a distribution of local excitations reminiscent, in some respects, of a thermal bath. We discuss how the mathematical tools developed herein may open new ways for the analysis of fundamental problems in local quantum field theory.

  11. Wellhead protection programs: Tools for local governments

    SciTech Connect

    Harvey, R.; Linquiti, P.

    1989-04-01

    The Technical Assistance Document (TAD) describes how localities can, as a part of a State Wellhead Protection Program, develop and implement effective techniques for the protection of ground water. The document emphasizes innovative wellhead protection methods that have been used by local communities, discusses combinations of programs that have worked well, and presents several factors that affect the success of local wellhead protection programs, such as budgetary constraints and legal issues.

  12. Speech enhancement using local spectral regularization

    NASA Astrophysics Data System (ADS)

    Sandoval-Ibarra, Yuma; Diaz-Ramirez, Victor H.; Kober, Vitaly; Diaz, Arnoldo

    2016-09-01

    A locally-adaptive algorithm for speech enhancement based on local spectral regularization is presented. The algorithm is able to retrieve a clean speech signal from a noisy signal using locally-adaptive signal processing. The proposed algorithm is able to increase the quality of a noisy signal in terms of objective metrics. Computer simulation results obtained with the proposed algorithm are presented and discussed in processing speech signals corrupted with additive noise.

  13. Tactile localization depends on stimulus intensity.

    PubMed

    Steenbergen, Peter; Buitenweg, Jan R; Trojan, Jörg; Veltink, Peter H

    2014-02-01

    Few experimental data are available about the influence of stimulus intensity on localization of cutaneous stimuli. The localization behavior of an individual as function of the veridical stimulus sites can be represented in the form of a perceptual map. It is unknown how the intensity of cutaneous stimuli influences these perceptual maps. We investigated the effect of stimulus intensity on trial-to-trial localization variability and on perceptual maps. We applied non-painful electrocutaneous stimuli of three different intensities through seven surface electrodes on the lower arm of healthy participants. They localized the stimuli on a tablet monitor mounted directly above their arm, on which a photograph of this arm was presented. The length of the arm over which the stimuli were localized was contracted when compared to the real electrode positions. This length increased toward veridical with increasing stimulus intensity. The trial-to-trial variance of the localizations dropped significantly with increasing intensity. Furthermore, localization biases of individual stimulus positions were shown to decrease with increasing stimulus intensity. We conclude that tactile stimuli are localized closer to veridical with increasing intensity in two respects: the localizations become more consistent and more accurate.

  14. Local, nonlocal quantumness and information theoretic measures

    NASA Astrophysics Data System (ADS)

    Agrawal, Pankaj; Sazim, Sk; Chakrabarty, Indranil; Pati, Arun K.

    2016-08-01

    It has been suggested that there may exist quantum correlations that go beyond entanglement. The existence of such correlations can be revealed by information theoretic quantities such as quantum discord, but not by the conventional measures of entanglement. We argue that a state displays quantumness, that can be of local and nonlocal origin. Information theoretic measures not only characterize the nonlocal quantumness, but also the local quantumness, such as the “local superposition”. This can be a reason, why such measures are nonzero, when there is no entanglement. We consider a generalized version of the Werner state to demonstrate the interplay of local quantumness, nonlocal quantumness and classical mixedness of a state.

  15. Balance and Ensemble Kalman Filter Localization Techniques

    DTIC Science & Technology

    2010-01-01

    For the serial EnSRF ( Whitaker and Hamill, 2002), localization by a distance-dependent function is performed upon BHT, where each element...localization in terms of balance and accuracy. Here, B-localization is employed with the EnSRF algorithm ( Whitaker and Hamill, 2002), whereas R- localization...can be specified as follows:       2 1 K K   1111 12 11       RB B B (A1) where Bij is the background covariance between

  16. 22 CFR 228.40 - Local procurement.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... available locally: (1) Utilities, including fuel for heating and cooking, waste disposal and trash... housing and office space; (4) Petroleum, oils and lubricants for operating vehicles and equipment;...

  17. Key Objectives for State and Local Governments

    EPA Pesticide Factsheets

    This page discusses key objectives for state and local governments when developing Clean Energy Financing Programs, including generating demand, encouraging private lenders, and long-term program sustainability.

  18. Functional Localization of Genetic Network Programming

    NASA Astrophysics Data System (ADS)

    Eto, Shinji; Hirasawa, Kotaro; Hu, Jinglu

    According to the knowledge of brain science, it is suggested that there exists cerebral functional localization, which means that a specific part of the cerebrum is activated depending on various kinds of information human receives. The aim of this paper is to build an artificial model to realize functional localization based on Genetic Network Programming (GNP), a new evolutionary computation method recently developed. GNP has a directed graph structure suitable for realizing functional localization. We studied the basic characteristics of the proposed system by making GNP work in a functionally localized way.

  19. Local anesthetics induce human renal cell apoptosis.

    PubMed

    Lee, H Thomas; Xu, Hua; Siegel, Cory D; Krichevsky, Igor E

    2003-01-01

    Renal cell apoptosis contributes significantly to the pathogenesis of acute renal failure. Local anesthetics induce apoptosis in neuronal and lymphocytic cell lines. We examined the effects of chronic (48 h) local anesthetic treatment (lidocaine, bupivacaine and tetracaine) on human proximal tubular (HK-2) cells. Apoptosis induction was assessed by detecting poly(ADP)-ribose polymerase fragmentation, caspase activation, terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining, DNA laddering and by cellular morphology. Cell death was quantified by measuring neutral red dye uptake and lactate dehydrogenase released into the cell culture medium. All 3 local anesthetics caused concentration-dependent cell death, induced HK-2 cell apoptosis and potentiated TNF-alpha induced apoptosis. Local anesthetics induced HK-2 cell apoptosis by activation of caspases 3, 6, 7, 8 and 9. ZVAD-fmk, a pan-caspase inhibitor, blocked the local anesthetic induced HK-2 cell apoptosis. Local anesthetics also inhibited the activities of anti-apoptotic kinases protein kinase B (Akt) and extracellular signal regulated mitrogen-activated protein kinase. Local anesthetic's pro-apoptotic effects are independent of sodium channel inhibition as tetrodotoxin, a selective voltage-gated sodium channel blocker, failed to mimic local anesthetic-mediated induction or potentiation of HK-2 cell apoptosis. We conclude that local anesthetics induce human renal cell apoptotic signaling by caspase activation and via inhibition of pro-survival signaling pathways.

  20. Tuning vibrational mode localization with frequency windowing

    NASA Astrophysics Data System (ADS)

    Cheng, Xiaolu; Talbot, Justin J.; Steele, Ryan P.

    2016-09-01

    Local-mode coordinates have previously been shown to be an effective starting point for anharmonic vibrational spectroscopy calculations. This general approach borrows techniques from localized-orbital machinery in electronic structure theory and generates a new set of spatially localized vibrational modes. These modes exhibit a well-behaved spatial decay of anharmonic mode couplings, which, in turn, allows for a systematic, a priori truncation of couplings and increased computational efficiency. Fully localized modes, however, have been found to lead to unintuitive mixtures of characteristic motions, such as stretches and bends, and accordingly large bilinear couplings. In this work, a very simple, tunable localization frequency window is introduced, in order to realize the transition from normal modes to fully localized modes. Partial localization can be achieved by localizing only pairs of modes within this traveling frequency window, which allows for intuitive interpretation of modes. The optimal window size is suggested to be a few hundreds of wave numbers, based on small- to medium-sized test systems, including water clusters and polypeptides. The new sets of partially localized coordinates retain their spatial coupling decay behavior while providing a reduced number of potential energy evaluations for convergence of anharmonic spectra.