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Sample records for abc transporter required

  1. Molecular and immunological analysis of an ABC transporter complex required for cytochrome c biogenesis.

    Goldman, B S; Beckman, D L; Bali, A; Monika, E M; Gabbert, K K; Kranz, R G

    1997-05-16

    The helABC genes are predicted to encode an ATP-binding cassette (ABC) transporter necessary for heme export for ligation in bacterial cytochrome c biogenesis. The recent discoveries of homologs of the helB and helC genes in plant mitochondrial genomes suggest this is a highly conserved transporter in prokaryotes and some eukaryotes with the HelB and HelC proteins comprising the transmembrane components. Molecular genetic analysis in the Gram-negative bacterium Rhodobacter capsulatus was used to show that the helABC and helDX genes are part of an operon linked to the secDF genes. To facilitate analysis of this transporter, strains with non-polar deletions in each gene, epitope and reporter-tagged HelABCD proteins, and antisera specific to the HelA and HelX proteins were generated. We directly demonstrate that this transporter is present in the cytoplasmic membrane as an HelABCD complex. The HelB and HelC but not HelD proteins are necessary for the binding and stability of the HelA protein, the cytoplasmic subunit containing the ATP-binding region. In addition we show that the HelA protein co-immunoprecipitates with either the HelC or HelD proteins. Thus, the HelABCD heme export complex is distinguished by the presence of four membrane-associated subunits and represents a unique subfamily of ABC transporters. PMID:9175857

  2. The predicted ABC transporter AbcEDCBA is required for type IV secretion system expression and lysosomal evasion by Brucella ovis.

    Teane M A Silva

    Full Text Available Brucella ovis is a major cause of reproductive failure in rams and it is one of the few well-described Brucella species that is not zoonotic. Previous work showed that a B. ovis mutant lacking a species-specific ABC transporterabcBA was attenuated in mice and was unable to survive in macrophages. The aim of this study was to evaluate the role of this ABC transporter during intracellular survival of B. ovis. In HeLa cells, B. ovis WT was able to survive and replicate at later time point (48 hpi, whereas an ΔabcBA mutant was attenuated at 24 hpi. The reduced survival of the ΔabcBA mutant was associated with a decreased ability to exclude the lysosomal marker LAMP1 from its vacuolar membrane, suggesting a failure to establish a replicative niche. The ΔabcBA mutant showed a reduced abundance of the Type IV secretion system (T4SS proteins VirB8 and VirB11 in both rich and acid media, when compared to WT B. ovis. However, mRNA levels of virB1, virB8, hutC, and vjbR were similar in both strains. These results support the notion that the ABC transporter encoded by abcEDCBA or its transported substrate acts at a post-transcriptional level to promote the optimal expression of the B. ovis T4SS within infected host cells.

  3. ABC Transporters, Atherosclerosis and Inflammation

    Fitzgerald, Michael L.; Mujawar, Zahedi; Tamehiro, Norimasa

    2010-01-01

    Atherosclerosis, driven by inflamed lipid-laden lesions, can occlude the coronary arteries and lead to myocardial infarction. This chronic disease is a major and expensive health burden. However, the body is able to mobilize and excrete cholesterol and other lipids, thus preventing atherosclerosis by a process termed reverse cholesterol transport (RCT). Insight into the mechanism of RCT has been gained by the study of two rare syndromes caused by the mutation of ABC transporter loci. In Tangi...

  4. Mapping the functional yeast ABC transporter interactome.

    Snider, Jamie; Hanif, Asad; Lee, Mid Eum; Jin, Ke; Yu, Analyn R; Graham, Chris; Chuk, Matthew; Damjanovic, Dunja; Wierzbicka, Marta; Tang, Priscilla; Balderes, Dina; Wong, Victoria; Jessulat, Matthew; Darowski, Katelyn D; San Luis, Bryan-Joseph; Shevelev, Igor; Sturley, Stephen L; Boone, Charles; Greenblatt, Jack F; Zhang, Zhaolei; Paumi, Christian M; Babu, Mohan; Park, Hay-Oak; Michaelis, Susan; Stagljar, Igor

    2013-09-01

    ATP-binding cassette (ABC) transporters are a ubiquitous class of integral membrane proteins of immense clinical interest because of their strong association with human disease and pharmacology. To improve our understanding of these proteins, we used membrane yeast two-hybrid technology to map the protein interactome of all of the nonmitochondrial ABC transporters in the model organism Saccharomyces cerevisiae and combined this data with previously reported yeast ABC transporter interactions in the BioGRID database to generate a comprehensive, integrated 'interactome'. We show that ABC transporters physically associate with proteins involved in an unexpectedly diverse range of functions. We specifically examine the importance of the physical interactions of ABC transporters in both the regulation of one another and in the modulation of proteins involved in zinc homeostasis. The interaction network presented here will be a powerful resource for increasing our fundamental understanding of the cellular role and regulation of ABC transporters. PMID:23831759

  5. Mapping the functional yeast ABC transporter interactome

    Snider, Jamie; Hanif, Asad; Lee, Mid Eum; Jin, Ke; Yu, Analyn R.; Graham, Chris; Chuk, Matthew; Damjanovic, Dunja; Wierzbicka, Marta; Tang, Priscilla; Balderes, Dina; Wong, Victoria; Jessulat, Matthew; Darowski, Katelyn D.; Luis, Bryan-Joseph San

    2013-01-01

    ABC transporters are a ubiquitous class of integral membrane proteins of immense clinical interest because of their strong association with human disease and pharmacology. To improve our understanding of these proteins, we used Membrane Yeast Two-Hybrid (MYTH) technology to map the protein interactome of all non-mitochondrial ABC transporters in the model organism Saccharomy cescerevisiae, and combined this data with previously reported yeast ABC transporter interactions in the BioGRID databa...

  6. ABC-B transporter genes in Dirofilaria immitis.

    Bourguinat, Catherine; Che, Hua; Mani, Thangadurai; Keller, Kathy; Prichard, Roger K

    2016-08-01

    Dirofilaria immitis is a filarial nematode causing infection and heartworm disease in dogs and other canids, cats, and occasionally in humans. Prevention with macrocyclic lactones (ML) is recommended during the mosquito transmission season. Recently, ML resistance has been reported. ABC-B transporter genes are thought to be involved in the mechanism of ML resistance in other nematodes. This study aimed to identify all the ABC-B transporter genes in D. immitis using as a reference the nDi.2.2 D. immitis whole genome, which is not completely annotated. Using bioinformatic tools and PCR amplification on pooled D. immitis genomic DNA and on pooled cDNA, nine ABC transporter genes including one pseudogene were characterized. Bioinformatic and phylogenetic analyses allowed identification of three P-glycoproteins (Pgps) (Dim-pgp-3 Dim-pgp-10, Dim-pgp-11), of two ABC-B half transporter genes (one ortholog of Cel-haf-4 and Cel-haf-9; and one ortholog of Cel-haf-1 and Cel-haf-3), of one ABC half transporter gene (ortholog of Cel-haf-5) that contained an ABC-C motif, and of one additional half transporter that would require functional study for characterization. The number of ABC-B transporter genes identified was lower than in Caenorhabditis elegans and Haemonchus contortus. Further studies are needed to understand their possible role in ML resistance in D. immitis. These ABC transporters constitute a base for ML resistance investigation in D. immitis and advance our understanding of the molecular biology of this parasite. PMID:27164440

  7. Fungal ABC transporter deletion and localization analysis.

    Kovalchuk, Andriy; Weber, Stefan S; Nijland, Jeroen G; Bovenberg, Roel A L; Driessen, Arnold J M

    2012-01-01

    Fungal cells are highly complex as their metabolism is compartmentalized harboring various types of subcellular organelles that are bordered by one or more membranes. Knowledge about the intracellular localization of transporter proteins is often required for the understanding of their biological function. Among different approaches available, the localization analysis based on the expression of GFP fusions is commonly used as a relatively fast and cost-efficient method that allows visualization of proteins of interest in both live and fixed cells. In addition, inactivation of transporter genes is an important tool to resolve their specific function. Here we provide a detailed protocol for the deletion and localization analysis of ABC transporters in the filamentous fungus Penicillium chrysogenum. It includes construction of expression plasmids, their transformation into fungal strains, cultivation of transformants, microscopy analysis, as well as additional protocols on staining of fungal cells with organelle-specific dyes like Hoechst 33342, MitoTracker DeepRed, and FM4-64. PMID:22183644

  8. Abc1: a new ABC transporter from the fission yeast Schizosaccharomyces pombe

    Christensen, P U; Davis, K; Nielsen, O; Davey, William John; Nielsen, Olaf

    1997-01-01

    We have isolated the abc1 gene from the fission yeast Schizosaccharomyces pombe. Sequence analysis suggests that the Abc1 protein is a member of the ABC superfamily of transporters and is composed of two structurally homologous halves, each consisting of a hydrophobic region of six transmembrane...

  9. Identification and functional characterization of Penicillium marneffei pleiotropic drug resistance transporters ABC1 and ABC2.

    Panapruksachat, Siribun; Iwatani, Shun; Oura, Takahiro; Vanittanakom, Nongnuch; Chindamporn, Ariya; Niimi, Kyoko; Niimi, Masakazu; Lamping, Erwin; Cannon, Richard D; Kajiwara, Susumu

    2016-07-01

    Penicilliosis caused by the dimorphic fungus Penicillium marneffei is an endemic, AIDS-defining illness and, after tuberculosis and cryptococcosis, the third most common opportunistic infection of AIDS patients in tropical Southeast Asia. Untreated, patients have poor prognosis; however, primary amphotericin B treatment followed by prolonged itraconazole prophylaxis is effective. To identify ATP-binding cassette (ABC) transporters that may play a role in potential multidrug resistance of P. marneffei, we identified and classified all 46 P. marneffei ABC transporters from the genome sequence. PmABC1 and PmABC2 were most similar to the archetype Candida albicans multidrug efflux pump gene CDR1. P. marneffei Abc1p (PmAbc1p) was functionally expressed in Saccharomyces cerevisiae, although at rather low levels, and correctly localized to the plasma membrane, causing cells to be fourfold to eightfold more resistant to azoles and many other xenobiotics than untransformed cells. P. marneffei Abc2p (PmAbc2p) was expressed at similarly low levels, but it had no efflux activity and did not properly localize to the plasma membrane. Interestingly, PmAbc1p mislocalized and lost its transport activity when cells were shifted to 37 °C. We conclude that expression of PmAbc1p in S. cerevisiae confers resistance to several xenobiotics indicating that PmAbc1p may be a multidrug efflux pump. PMID:26782644

  10. Isolation and Characterization of the Colletotrichum acutatum ABC Transporter CaABC1

    Suyoung Kim

    2014-12-01

    Full Text Available Fungi tolerate exposure to various abiotic stresses, including cytotoxic compounds and fungicides, via their ATP-driven efflux pumps belonging to ATP-binding cassette (ABC transporters. To clarify the molecular basis of interaction between the fungus and various abiotic stresses including fungicides, we constructed a cDNA library from germinated conidia of Colletotrichum acutatum, a major anthracnose pathogen of pepper (Capsicum annum L.. Over 1,000 cDNA clones were sequenced, of which single clone exhibited significant nucleotide sequence homology to ABC transporter genes. We isolated three fosmid clones containing the C. acutatum ABC1 (CaABC1 gene in full-length from genomic DNA library screening. The CaABC1 gene consists of 4,059 bp transcript, predicting a 1,353-aa protein. The gene contains the typical ABC signature and Walker A and B motifs. The 5′-flanking region contains a CAAT motif, a TATA box, and a Kozak region. Phylogenetic and structural analysis suggested that the CaABC1 is a typical ABC transporter gene highly conserved in various fungal species, as well as in Chromista, Metazoans, and Viridiplantae. We also found that CaABC1 was up-regulated during conidiation and a minimal medium condition. Moreover, CaABC1 was induced in iprobenfos, kresoxim-methyl, thiophanate-methyl, and hygromycin B. These results demonstrate that CaABC1 is necessary for conidiation, abiotic stress, and various fungicide resistances. These results will provide the basis for further study on the function of ABC transporter genes in C. acutatum.

  11. The N-terminal extension domain of the C. elegans half-molecule ABC transporter, HMT-1, is required for protein-protein interactions and function.

    Sungjin Kim

    Full Text Available BACKGROUND: Members of the HMT-1 (heavy metal tolerance factor 1 subfamily of the ATP-binding cassette (ABC transporter superfamily detoxify heavy metals and have unique topology: they are half-molecule ABC transporters that, in addition to a single transmembrane domain (TMD1 and a single nucleotide-binding domain (NBD1, possess a hydrophobic NH2-terminal extension (NTE. These structural features distinguish HMTs from other ABC transporters in different species including Drosophila and humans. Functional ABC transporters, however, are comprised of at least four-domains (two TMDs and two NDBs formed from either a single polypeptide or by the association of two or four separate subunits. Whether HMTs act as oligomers and what role the NTE domain plays in their function have not been determined. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we examined the oligomeric status of Caenorhabditis elegans HMT-1 and the functional significance of its NTE using gel-filtration chromatography in combination with the mating-based split-ubiquitin yeast two-hybrid system (mbSUS and functional in vivo assays. We found that HMT-1 exists in a protein complex in C. elegans. Studies in S. cerevisiae showed that HMT-1 at a minimum homodimerizes and that oligomerization is essential for HMT-1 to confer cadmium tolerance. We also established that the NTE domain plays an important structural and functional role: it is essential for HMT-1 oligomerization and Cd-detoxification function. However, the NTE itself was not sufficient for oligomerization suggesting that multiple structural features of HMT-1 must associate to form a functional transporter. CONCLUSIONS: The prominence of heavy metals as environmental toxins and the remarkable conservation of HMT-1 structural architecture and function in different species reinforce the value of continued studies of HMT-1 in model systems for identifying functional domains in HMT1 of humans.

  12. How heterogeneous is the involvement of ABC transporters against insecticides?

    Porretta, Daniele; Epis, Sara; Mastrantonio, Valentina; Ferrari, Marco; Bellini, Romeo; Favia, Guido; Urbanelli, Sandra

    2016-05-01

    Understanding the molecular mechanisms underlying cellular defense against xenobiotic compounds is a main research issue in medical and veterinary entomology, as insecticide/acaricide resistance is a major threat in the control of arthropods. ABC transporters are recognized as a component of the detoxifying mechanism in arthropods. We investigated the possible involvement of ABC transporters in defense to the organophosphate insecticide temephos in the malarial vector Anopheles stephensi. We performed bioassays on larvae of An. stephensi, using insecticide alone and in combination with ABC-transporter inhibitors, to assess synergism between these compounds. Next, we investigated the expression profiles of six ABC transporter genes in larvae exposed to temephos. Surprisingly, neither bioassays nor gene expression analyses provided any evidence for a major role of ABC transporters in defense against temephos in An. stephensi. We thus decided to review existing literature to generate a record of other studies that failed to reveal a role for ABC transporters against particular insecticides/acaricides. A review of the scientific literature led to the recovery of 569 papers about ABC transporters; among these, 50 involved arthropods, and 10 reported negative results. Our study on An. stephensi and accompanying literature review highlight the heterogeneity that exists in ABC transporter involvement in defense/resistance mechanisms in arthropods. PMID:26855383

  13. Human ATP-binding cassette (ABC transporter family

    Vasiliou Vasilis

    2009-04-01

    Full Text Available Abstract There exist four fundamentally different classes of membrane-bound transport proteins: ion channels; transporters; aquaporins; and ATP-powered pumps. ATP-binding cassette (ABC transporters are an example of ATP-dependent pumps. ABC transporters are ubiquitous membrane-bound proteins, present in all prokaryotes, as well as plants, fungi, yeast and animals. These pumps can move substrates in (influx or out (efflux of cells. In mammals, ABC transporters are expressed predominantly in the liver, intestine, blood-brain barrier, blood-testis barrier, placenta and kidney. ABC proteins transport a number of endogenous substrates, including inorganic anions, metal ions, peptides, amino acids, sugars and a large number of hydrophobic compounds and metabolites across the plasma membrane, and also across intracellular membranes. The human genome contains 49 ABC genes, arranged in eight subfamilies and named via divergent evolution. That ABC genes are important is underscored by the fact that mutations in at least I I of these genes are already known to cause severe inherited diseases (eg cystic fibrosis and X-linked adrenoleukodystrophy [X-ALD]. ABC transporters also participate in the movement of most drugs and their metabolites across cell surface and cellular organelle membranes; thus, defects in these genes can be important in terms of cancer therapy, pharmacokinetics and innumerable pharmacogenetic disorders.

  14. The role of ABCG-type ABC transporters in phytohormone transport

    Borghi, Lorenzo; Kang, Joohyun; Ko, Donghwi; Lee, Youngsook; Martinoia, Enrico

    2015-01-01

    Plant hormones (phytohormones) integrate endogenous and exogenous signals thus synchronizing plant growth with environmental and developmental changes. Similar to animals, phytohormones have distinct source and target tissues, hence controlled transport and focused targeting are required for their functions. Many evidences accumulated in the last years about the regulation of long-distance and directional transport of phytohormones. ATP-binding cassette (ABC) transporters turned out to play m...

  15. ABC Transporters and their Role in Nucleoside and Nucleotide Drug Resistance

    Fukuda, Yu; Schuetz, John D.

    2012-01-01

    ATP-binding cassette (ABC) transporters confer drug resistance against a wide range of chemotherapeutic agents, including nucleoside and nucleotide based drugs. While nucleoside based drugs have been used for many years in the treatment of solid and hematological malignancies as well as viral and autoimmune diseases, the potential contribution of ABC transporters has only recently been recognized. This neglect is likely because activation of nucleoside derivatives require an initial carrier-m...

  16. Export of recombinant proteins in Escherichia coli using ABC transporter with an attached lipase ABC transporter recognition domain (LARD

    Moon Yuseok

    2009-01-01

    Full Text Available Abstract Background ATP binding cassette (ABC transporter secretes the protein through inner and outer membranes simultaneously in gram negative bacteria. Thermostable lipase (TliA of Pseudomonas fluorescens SIK W1 is secreted through the ABC transporter. TliA has four glycine-rich repeats (GGXGXD in its C-terminus, which appear in many ABC transporter-secreted proteins. From a homology model of TliA derived from the structure of P. aeruginosa alkaline protease (AprA, lipase ABC transporter domains (LARDs were designed for the secretion of fusion proteins. Results The LARDs included four glycine-rich repeats comprising a β-roll structure, and were added to the C-terminus of test proteins. Either Pro-Gly linker or Factor Xa site was added between fusion proteins and LARDs. We attached different length of LARDs such as LARD0, LARD1 or whole TliA (the longest LARD to three types of proteins; green fluorescent protein (GFP, epidermal growth factor (EGF and cytoplasmic transduction peptide (CTP. These fusion proteins were expressed in Escherichia coli together with ABC transporter of either P. fluorescens or Erwinia chrysanthemi. Export of fusion proteins with the whole TliA through the ABC transporter was evident on the basis of lipase enzymatic activity. Upon supplementation of E. coli with ABC transporter, GFP-LARDs and EGF-LARDs were excreted into the culture supernatant. Conclusion The LARDs or whole TliA were attached to C-termini of model proteins and enabled the export of the model proteins such as GFP and EGF in E. coli supplemented with ABC transporter. These results open the possibility for the extracellular production of recombinant proteins in Pseudomonas using LARDs or TliA as a C-terminal signal sequence.

  17. Asymmetric switching in a homodimeric ABC transporter: a simulation study.

    Jussi Aittoniemi

    2010-04-01

    Full Text Available ABC transporters are a large family of membrane proteins involved in a variety of cellular processes, including multidrug and tumor resistance and ion channel regulation. Advances in the structural and functional understanding of ABC transporters have revealed that hydrolysis at the two canonical nucleotide-binding sites (NBSs is co-operative and non-simultaneous. A conserved core architecture of bacterial and eukaryotic ABC exporters has been established, as exemplified by the crystal structure of the homodimeric multidrug exporter Sav1866. Currently, it is unclear how sequential ATP hydrolysis arises in a symmetric homodimeric transporter, since it implies at least transient asymmetry at the NBSs. We show by molecular dynamics simulation that the initially symmetric structure of Sav1866 readily undergoes asymmetric transitions at its NBSs in a pre-hydrolytic nucleotide configuration. MgATP-binding residues and a network of charged residues at the dimer interface are shown to form a sequence of putative molecular switches that allow ATP hydrolysis only at one NBS. We extend our findings to eukaryotic ABC exporters which often consist of two non-identical half-transporters, frequently with degeneracy substitutions at one of their two NBSs. Interestingly, many residues involved in asymmetric conformational switching in Sav1866 are substituted in degenerate eukaryotic NBS. This finding strengthens recent suggestions that the interplay of a consensus and a degenerate NBS in eukaroytic ABC proteins pre-determines the sequence of hydrolysis at the two NBSs.

  18. ABC transporters from Botrytis cinerea in biotic and abiotic interactions

    Schoonbeek, H.

    2004-01-01

    Botrytis cinereais the causal agent of grey mould disease on a wide variety of crop plants. It is relatively insensitive to natural and synthetic fungitoxic compounds. This thesis describes how ABC (ATP-binding cassette) transporters contribute to protection by actively secre

  19. The ABCs of Candida albicans Multidrug Transporter Cdr1.

    Prasad, Rajendra; Banerjee, Atanu; Khandelwal, Nitesh Kumar; Dhamgaye, Sanjiveeni

    2015-12-01

    In the light of multidrug resistance (MDR) among pathogenic microbes and cancer cells, membrane transporters have gained profound clinical significance. Chemotherapeutic failure, by far, has been attributed mainly to the robust and diverse array of these proteins, which are omnipresent in every stratum of the living world. Candida albicans, one of the major fungal pathogens affecting immunocompromised patients, also develops MDR during the course of chemotherapy. The pivotal membrane transporters that C. albicans has exploited as one of the strategies to develop MDR belongs to either the ATP binding cassette (ABC) or the major facilitator superfamily (MFS) class of proteins. The ABC transporter Candida drug resistance 1 protein (Cdr1p) is a major player among these transporters that enables the pathogen to outplay the battery of antifungals encountered by it. The promiscuous Cdr1 protein fulfills the quintessential need of a model to study molecular mechanisms of multidrug transporter regulation and structure-function analyses of asymmetric ABC transporters. In this review, we cover the highlights of two decades of research on Cdr1p that has provided a platform to study its structure-function relationships and regulatory circuitry for a better understanding of MDR not only in yeast but also in other organisms. PMID:26407965

  20. Protein secretion by hybrid bacterial ABC-transporters: specific functions of the membrane ATPase and the membrane fusion protein.

    Binet, R; Wandersman, C

    1995-01-01

    The Erwinia chrysanthemi metalloprotease C and the Serratia marcescens haem acquisition protein HasA are both secreted from Gram-negative bacteria by a signal peptide-independent pathway which requires a C-terminal secretion signal and a specific ABC-transporter made up of three proteins: a membrane ATPase (the ABC-protein), a second inner membrane component belonging to the membrane fusion protein family and an outer membrane polypeptide. HasA and protease C transporters are homologous altho...

  1. Know your ABCs: Characterization and gene expression dynamics of ABC transporters in the polyphagous herbivore Helicoverpa armigera.

    Bretschneider, Anne; Heckel, David G; Vogel, Heiko

    2016-05-01

    Polyphagous insect herbivores are adapted to many different secondary metabolites of their host plants. However, little is known about the role of ATP-binding cassette (ABC) transporters, a multigene family involved in detoxification processes. To study the larval response of the generalist Helicoverpa armigera (Lepidoptera) and the putative role of ABC transporters, we performed developmental assays on artificial diet supplemented with secondary metabolites from host plants (atropine-scopolamine, nicotine and tomatine) and non-host plants (taxol) in combination with a replicated RNAseq experiment. A maximum likelihood phylogeny identified the subfamily affiliations of the ABC transporter sequences. Larval performance was equal on the atropine-scopolamine diet and the tomatine diet. For the latter we could identify a treatment-specific upregulation of five ABC transporters in the gut. No significant developmental difference was detected between larvae fed on nicotine or taxol. This was also mirrored in the upregulation of five ABC transporters when fed on either of the two diets. The highest number of differentially expressed genes was recorded in the gut samples in response to feeding on secondary metabolites. Our results are consistent with the expectation of a general detoxification response in a polyphagous herbivore. This is the first study to characterize the multigene family of ABC transporters and identify gene expression changes across different developmental stages and tissues, as well as the impact of secondary metabolites in the agricultural pest H. armigera. PMID:26951878

  2. Conformational coupling of the nucleotide-binding and the transmembrane domains in ABC transporters.

    Wen, Po-Chao; Tajkhorshid, Emad

    2011-08-01

    Basic architecture of ABC transporters includes two transmembrane domains (TMDs) and two nucleotide-binding domains (NBDs). Although the transport process takes place in the TMDs, which provide the substrate translocation pathway across the cell membrane and control its accessibility between the two sides of the membrane, the energy required for the process is provided by conformational changes induced in the NBDs by binding and hydrolysis of ATP. Nucleotide-dependent conformational changes in the NBDs, therefore, need to be coupled to structural changes in the TMDs. Using molecular dynamics simulations, we have investigated the structural elements involved in the conformational coupling between the NBDs and the TMDs in the Escherichia coli maltose transporter, an ABC importer for which an intact structure is available both in inward-facing and outward-facing conformations. The prevailing model of coupling is primarily based on a single structural motif, known as the coupling helices, as the main structural element for the NBD-TMD coupling. Surprisingly, we find that in the absence of the NBDs the coupling helices can be conformationally decoupled from the rest of the TMDs, despite their covalent connection. That is, the structural integrity of the coupling helices and their tight coupling to the core of the TMDs rely on the contacts provided by the NBDs. Based on the conformational and dynamical analysis of the simulation trajectories, we propose that the core coupling elements in the maltose transporter involve contributions from several structural motifs located at the NBD-TMD interface, namely, the EAA loops from the TMDs, and the Q-loop and the ENI motifs from the NBDs. These three structural motifs in small ABC importers show a high degree of correlation in motion and mediate the necessary conformational coupling between the core of TMDs and the helical subdomains of NBDs. A comprehensive analysis of the structurally known ABC transporters shows a high degree

  3. A reciprocating twin-channel model for ABC transporters.

    Jones, Peter M; George, Anthony M

    2014-08-01

    ABC transporters comprise a large, diverse, and ubiquitous superfamily of membrane active transporters. Their core architecture is a dimer of dimers, comprising two transmembrane (TM) domains that bind substrate, and two ATP-binding cassettes, which use the cell's energy currency to couple substrate translocation to ATP hydrolysis. Despite the availability of over a dozen resolved structures and a wealth of biochemical and biophysical data, this field is bedeviled by controversy and long-standing mechanistic questions remain unresolved. The prevailing paradigm for the ABC transport mechanism is the Switch Model, in which the ATP-binding cassettes dimerize upon binding two ATP molecules, and thence dissociate upon sequential ATP hydrolysis. This cycle of nucleotide-binding domain (NBD) dimerization and dissociation is coupled to a switch between inward- or outward facing conformations of a single TM channel; this alternating access enables substrate binding on one face of the membrane and its release at the other. Notwithstanding widespread acceptance of the Switch Model, there is substantial evidence that the NBDs do not separate very much, if at all, and thus physical separation of the ATP cassettes observed in crystallographic structures may be an artefact. An alternative Constant Contact Model has been proposed, in which ATP hydrolysis occurs alternately at the two ATP-binding sites, with one of the sites remaining closed and containing occluded nucleotide at all times. In this model, the cassettes remain in contact and the active sites swing open in an alternately seesawing motion. Whilst the concept of NBD association/dissociation in the Switch Model is naturally compatible with a single alternating-access channel, the asymmetric functioning proposed by the Constant Contact model suggests an alternating or reciprocating function in the TMDs. Here, a new model for the function of ABC transporters is proposed in which the sequence of ATP binding, hydrolysis, and

  4. Inhibition of ABC transporters associated with multidrug resistance

    Egger, Michael

    2010-01-01

    The first part of this dissertation (chapters 1 and 2) deals with the inhibition of the ABC transporters ABCB1 (p-glycoprotein) and ABCG2 (breast cancer resistance protein). Less lipophilic and better water soluble analogues of the known ABCB1 inhibitor tariquidar were synthesized from one central building block via Cu(I)-catalyzed N/O-arylation reactions. These compounds were tested for their inhibitory activity against the ABCB1 transporter in a flow cytometric calcein-AM efflux assay and a...

  5. Mutations in the Arabidopsis Peroxisomal ABC Transporter COMATOSE Allow Differentiation between Multiple Functions In Planta: Insights from an Allelic Series

    Dietrich, Daniela; Schmuths, Heike; Lousa, Carine De Marcos; Baldwin, Jocelyn M.; Baldwin, Stephen A.; Baker, Alison; Theodoulou, Frederica L; Holdsworth, Michael J

    2009-01-01

    COMATOSE (CTS), the Arabidopsis homologue of human Adrenoleukodystrophy protein (ALDP), is required for import of substrates for peroxisomal β-oxidation. A new allelic series and a homology model based on the bacterial ABC transporter, Sav1866, provide novel insights into structure-function relations of ABC subfamily D proteins. In contrast to ALDP, where the majority of mutations result in protein absence from the peroxisomal membrane, all CTS mutants produced stable protein. Mutation of con...

  6. The Twisted Dwarf's ABC: How Immunophilins Regulate Auxin Transport

    Bailly, Aurélien; Sovero, Valpuri; Geisler, Markus

    2006-01-01

    There is increasing evidence that immunophilins function as key regulators of plant development. One of the best investigated members, the multi-domain FKBP TWISTED DWARF1 (TWD1)/FKBP42, has been shown to reside on both the vacuolar and plasma membranes where it interacts in mirror image with two pairs of ABC transporters, MRP1/ MRP2 and PGP1/PGP19(MDR1), respectively. Twisted dwarf1 and pgp1/pgp19 mutants display strongly overlapping phenotypes, including reduction and disorientation of grow...

  7. Toward Determining ATPase Mechanism in ABC Transporters: Development of the Reaction Path–Force Matching QM/MM Method

    Zhou, Y.; Ojeda-May, P.; Nagaraju, M.; Pu, J.

    2016-01-01

    Adenosine triphosphate (ATP)-binding cassette (ABC) transporters are ubiquitous ATP-dependent membrane proteins involved in translocations of a wide variety of substrates across cellular membranes. To understand the chemomechanical coupling mechanism as well as functional asymmetry in these systems, a quantitative description of how ABC transporters hydrolyze ATP is needed. Complementary to experimental approaches, computer simulations based on combined quantum mechanical and molecular mechanical (QM/MM) potentials have provided new insights into the catalytic mechanism in ABC transporters. Quantitatively reliable determination of the free energy requirement for enzymatic ATP hydrolysis, however, requires substantial statistical sampling on QM/MM potential. A case study shows that brute force sampling of ab initio QM/MM (AI/MM) potential energy surfaces is computationally impractical for enzyme simulations of ABC transporters. On the other hand, existing semiempirical QM/MM (SE/MM) methods, although affordable for free energy sampling, are unreliable for studying ATP hydrolysis. To close this gap, a multiscale QM/MM approach named reaction path–force matching (RP–FM) has been developed. In RP–FM, specific reaction parameters for a selected SE method are optimized against AI reference data along reaction paths by employing the force matching technique. The feasibility of the method is demonstrated for a proton transfer reaction in the gas phase and in solution. The RP–FM method may offer a general tool for simulating complex enzyme systems such as ABC transporters. PMID:27498639

  8. Toward Determining ATPase Mechanism in ABC Transporters: Development of the Reaction Path-Force Matching QM/MM Method.

    Zhou, Y; Ojeda-May, P; Nagaraju, M; Pu, J

    2016-01-01

    Adenosine triphosphate (ATP)-binding cassette (ABC) transporters are ubiquitous ATP-dependent membrane proteins involved in translocations of a wide variety of substrates across cellular membranes. To understand the chemomechanical coupling mechanism as well as functional asymmetry in these systems, a quantitative description of how ABC transporters hydrolyze ATP is needed. Complementary to experimental approaches, computer simulations based on combined quantum mechanical and molecular mechanical (QM/MM) potentials have provided new insights into the catalytic mechanism in ABC transporters. Quantitatively reliable determination of the free energy requirement for enzymatic ATP hydrolysis, however, requires substantial statistical sampling on QM/MM potential. A case study shows that brute force sampling of ab initio QM/MM (AI/MM) potential energy surfaces is computationally impractical for enzyme simulations of ABC transporters. On the other hand, existing semiempirical QM/MM (SE/MM) methods, although affordable for free energy sampling, are unreliable for studying ATP hydrolysis. To close this gap, a multiscale QM/MM approach named reaction path-force matching (RP-FM) has been developed. In RP-FM, specific reaction parameters for a selected SE method are optimized against AI reference data along reaction paths by employing the force matching technique. The feasibility of the method is demonstrated for a proton transfer reaction in the gas phase and in solution. The RP-FM method may offer a general tool for simulating complex enzyme systems such as ABC transporters. PMID:27498639

  9. The multidrug-resistant phenotype associated with overexpression of the new ABC half-transporter, MXR (ABCG2)

    Litman, Thomas; Brangi, M; Hudson, E;

    2000-01-01

    Mechanisms of drug resistance other than P-glycoprotein are of increasing interest as the list of newly identified members of the ABC transport family has grown. We sought to characterize the phenotype of the newly discovered ABC transporter encoded by the mitoxantrone resistance gene, MXR, also...... known as ABCP1 or BCRP. The pharmacodynamics of mitoxantrone and 12 other fluorescent drugs were evaluated by confocal microscopy in four multidrug-resistant human colon (S1) and breast (MCF-7) cancer cell lines. We utilized two sublines, MCF-7 AdVp3000 and S1-M1-80, and detected overexpression of MXR...... studies suggest that the ABC half-transporter, MXR, is a potent, new mechanism for conferring multiple drug resistance. Definition of its mechanism of transport and its role in clinical oncology is required....

  10. The maize glossy13 gene, cloned via BSR-Seq and Seq-walking encodes a putative ABC transporter required for the normal accumulation of epicuticular waxes.

    Li Li

    Full Text Available Aerial plant surfaces are covered by epicuticular waxes that among other purposes serve to control water loss. Maize glossy mutants originally identified by their "glossy" phenotypes exhibit alterations in the accumulation of epicuticular waxes. By combining data from a BSR-Seq experiment and the newly developed Seq-Walking technology, GRMZM2G118243 was identified as a strong candidate for being the glossy13 gene. The finding that multiple EMS-induced alleles contain premature stop codons in GRMZM2G118243, and the one knockout allele of gl13, validates the hypothesis that gene GRMZM2G118243 is gl13. Consistent with this, GRMZM2G118243 is an ortholog of AtABCG32 (Arabidopsis thaliana, HvABCG31 (barley and OsABCG31 (rice, which encode ABCG subfamily transporters involved in the trans-membrane transport of various secondary metabolites. We therefore hypothesize that gl13 is involved in the transport of epicuticular waxes onto the surfaces of seedling leaves.

  11. Antitubercular Agent Delamanid and Metabolites as Substrates and Inhibitors of ABC and Solute Carrier Transporters.

    Sasabe, Hiroyuki; Shimokawa, Yoshihiko; Shibata, Masakazu; Hashizume, Kenta; Hamasako, Yusuke; Ohzone, Yoshihiro; Kashiyama, Eiji; Umehara, Ken

    2016-06-01

    Delamanid (Deltyba, OPC-67683) is the first approved drug in a novel class of nitro-dihydro-imidazooxazoles developed for the treatment of multidrug-resistant tuberculosis. Patients with tuberculosis require treatment with multiple drugs, several of which have known drug-drug interactions. Transporters regulate drug absorption, distribution, and excretion; therefore, the inhibition of transport by one agent may alter the pharmacokinetics of another, leading to unexpected adverse events. Therefore, it is important to understand how delamanid affects transport activity. In the present study, the potencies of delamanid and its main metabolites as the substrates and inhibitors of various transporters were evaluated in vitro Delamanid was not transported by the efflux ATP-binding cassette (ABC) transporters P-glycoprotein (P-gp; MDR1/ABCB1) and breast cancer resistance protein (BCRP/ABCG2), solute carrier (SLC) transporters, organic anion-transporting polypeptides, or organic cation transporter 1. Similarly, metabolite 1 (M1) was not a substrate for any of these transporters except P-gp. Delamanid showed no inhibitory effect on ABC transporters MDR1, BCRP, and bile salt export pump (BSEP; ABCB11), SLC transporters, or organic anion transporters. M1 and M2 inhibited P-gp- and BCRP-mediated transport but did so only at the 50% inhibitory concentrations (M1, 4.65 and 5.71 μmol/liter, respectively; M2, 7.80 and 6.02 μmol/liter, respectively), well above the corresponding maximum concentration in plasma values observed following the administration of multiple doses in clinical trials. M3 and M4 did not affect the activities of any of the transporters tested. These in vitro data suggest that delamanid is unlikely to have clinically relevant interactions with drugs for which absorption and disposition are mediated by this group of transporters. PMID:27021329

  12. Functional analysis of the ATP-binding cassette (ABC transporter gene family of Tribolium castaneum

    Broehan Gunnar

    2013-01-01

    Full Text Available Abstract Background The ATP-binding cassette (ABC transporters belong to a large superfamily of proteins that have important physiological functions in all living organisms. Most are integral membrane proteins that transport a broad spectrum of substrates across lipid membranes. In insects, ABC transporters are of special interest because of their role in insecticide resistance. Results We have identified 73 ABC transporter genes in the genome of T. castaneum, which group into eight subfamilies (ABCA-H. This coleopteran ABC family is significantly larger than those reported for insects in other taxonomic groups. Phylogenetic analysis revealed that this increase is due to gene expansion within a single clade of subfamily ABCC. We performed an RNA interference (RNAi screen to study the function of ABC transporters during development. In ten cases, injection of double-stranded RNA (dsRNA into larvae caused developmental phenotypes, which included growth arrest and localized melanization, eye pigmentation defects, abnormal cuticle formation, egg-laying and egg-hatching defects, and mortality due to abortive molting and desiccation. Some of the ABC transporters we studied in closer detail to examine their role in lipid, ecdysteroid and eye pigment transport. Conclusions The results from our study provide new insights into the physiological function of ABC transporters in T. castaneum, and may help to establish new target sites for insect control.

  13. Plant ABC Transporters Enable Many Unique Aspects of a Terrestrial Plant's Lifestyle.

    Hwang, Jae-Ung; Song, Won-Yong; Hong, Daewoong; Ko, Donghwi; Yamaoka, Yasuyo; Jang, Sunghoon; Yim, Sojeong; Lee, Eunjung; Khare, Deepa; Kim, Kyungyoon; Palmgren, Michael; Yoon, Hwan Su; Martinoia, Enrico; Lee, Youngsook

    2016-03-01

    Terrestrial plants have two to four times more ATP-binding cassette (ABC) transporter genes than other organisms, including their ancestral microalgae. Recent studies found that plants harboring mutations in these transporters exhibit dramatic phenotypes, many of which are related to developmental processes and functions necessary for life on dry land. These results suggest that ABC transporters multiplied during evolution and assumed novel functions that allowed plants to adapt to terrestrial environmental conditions. Examining the literature on plant ABC transporters from this viewpoint led us to propose that diverse ABC transporters enabled many unique and essential aspects of a terrestrial plant's lifestyle, by transporting various compounds across specific membranes of the plant. PMID:26902186

  14. Two molybdate/tungstate ABC transporters that interact very differently with their substrate binding proteins

    Vigonsky, Elena; Ovcharenko, Elena; Lewinson, Oded

    2013-01-01

    In all kingdoms of life, ATP Binding Cassette (ABC) transporters participate in many physiological and pathological processes. Despite the diversity of their functions, they have been considered to operate by a largely conserved mechanism. One deviant is the vitamin B12 transporter BtuCD that has been shown to operate by a distinct mechanism. However, it is unknown if this deviation is an exotic example, perhaps arising from the nature of the transported moiety. Here we compared two ABC impor...

  15. An ambiguous interface – on the transport mechanism of the ABC transport complex TAP

    Großmann, Nina

    2012-01-01

    The adaptive immune system protects against daily infections and malignant transformation. In this, the translocation of antigenic peptides by the transporter associated with antigen processing (TAP) into the ER lumen is an essential step in the antigen presentation by MHC I molecules. The heterodimeric ATP-binding cassette transporter (ABC) TAP consist of the two halftransporters TAP1 and TAP2. Each monomer contains an N-terminal transmembrane domain (TMD) and a conserved C-terminal nucleoti...

  16. Functional analysis of candidate ABC transporter proteins for sitosterol transport

    Albrecht, C; Elliott, J I; Sardini, A;

    2002-01-01

    protein (Bcrp; Abcg2) and the bile salt export pump (Bsep; Abcb11) was assessed using several assays. Unexpectedly, none of the candidate proteins mediated significant sitosterol transport. This has implications for the pathology of sitosterolemia. In addition, the data suggest that otherwise broad...

  17. Homologous Expression of the Lipase and ABC Transporter Gene Cluster, tliDEFA, Enhances Lipase Secretion in Pseudomonas spp.

    Ahn, Jung Hoon; Pan, Jae Gu; Rhee, Joon Shick

    2001-01-01

    The ABC transporter TliDEF was found to be an efficient secretory apparatus for extracellular lipase TliA in Pseudomonas fluorescens. For the enhanced secretion of the lipase, we tried to coexpress tliA and tliDEF in various Pseudomonas species. Whereas the coexpression of tliA and tliDEF was required for the lipase secretion in P. fragi, the expression of tliA was sufficient for the lipase secretion in P. fluorescens, P. syringae, and P. putida, indicating the existence of compatible ABC tra...

  18. Glycolysis Inhibition Inactivates ABC Transporters to Restore Drug Sensitivity in Malignant Cells

    Ayako Nakano; Daisuke Tsuji; Hirokazu Miki; Qu Cui; Salah Mohamed El Sayed; Akishige Ikegame; Asuka Oda; Hiroe Amou; Shingen Nakamura; Takeshi Harada; Shiro Fujii; Kumiko Kagawa; Kyoko Takeuchi; Akira Sakai; Shuji Ozaki

    2011-01-01

    Cancer cells eventually acquire drug resistance largely via the aberrant expression of ATP-binding cassette (ABC) transporters, ATP-dependent efflux pumps. Because cancer cells produce ATP mostly through glycolysis, in the present study we explored the effects of inhibiting glycolysis on the ABC transporter function and drug sensitivity of malignant cells. Inhibition of glycolysis by 3-bromopyruvate (3BrPA) suppressed ATP production in malignant cells, and restored the retention of daunorubic...

  19. Influence of detergents on the activity of the ABC transporter LmrA

    Infed, Nacera; Hanekop, Nils; Driessen, Arnold J. M.; Smits, Sander H. J.; Schmitt, Lutz

    2011-01-01

    The ABC transporter LmrA from Lactococcus lactis has been intensively studied and a role in multidrug resistance was proposed. Here, we performed a comprehensive detergent screen to analyze the impact of detergents for a successful solubilization, purification and retention of functional properties of this ABC transporter. Our screen revealed the preference of LmrA for zwitterionic detergents. In detergent solution, LmrA purified with FC-16 was highly active with respect to ATPase activity, w...

  20. Mechanism of ABC transporters: A molecular dynamics simulation of a well characterized nucleotide-binding subunit

    Peter M Jones; Anthony M George

    2002-01-01

    ATP-binding cassette (ABC) transporters are membrane-bound molecular pumps that form one of the largest of all protein families. Several of them are central to phenomena of biomedical interest, including cystic fibrosis and resistance to chemotherapeutic drugs. ABC transporters share a common architecture comprising two hydrophilic nucleotide-binding domains (NBDs) and two hydrophobic transmembrane domains (TMDs) that form the substrate pathway across the membrane. The conformational changes ...

  1. Interaction of ABC multidrug transporters with anticancer protein kinase inhibitors: substrates and/or inhibitors?

    Hegedus, Csilla; Ozvegy-Laczka, Csilla; Szakács, Gergely; Sarkadi, Balázs

    2009-05-01

    Protein kinase inhibitors (PKI) are becoming key agents in modern cancer chemotherapy, and combination of PKIs with classical chemotherapeutic drugs may help to overcome currently untreatable metastatic cancers. Since chemotherapy resistance is a recurrent problem, mechanisms of resistance should be clarified in order to help further drug development. Here we suggest that in addition to PKI resistance based on altered target structures, the active removal of these therapeutic agents by the MDR-ABC transporters should also be considered as a major cause of clinical resistance. We discuss the occurring systemic and cellular mechanisms, which may hamper PKI efficiency, and document the role of selected MDR-ABC transporters in these phenomena through their interactions with these anticancer agents. Moreover, we suggest that PKI interactions with ABC transporters may modulate overall drug metabolism, including the fate of diverse, chemically or target-wise unrelated drugs. These effects are based on multiple forms of MDR-ABC transporter interaction with PKIs, as these compounds may be both substrates and/or inhibitors of an ABC transporter. We propose that these interactions should be carefully considered in clinical application, and a combined MDR-ABC transporter and PKI effect may bring a major advantage in future drug development. PMID:19442047

  2. Phenotypic profiling of ABC transporter coding genes in Myxococcus xanthus

    RoyDWelch

    2014-07-01

    Full Text Available Information about a gene sometimes can be deduced by examining the impact of its mutation on phenotype. However, the genome-scale utility of the method is limited because, for nearly all model organisms, the majority of mutations result in little or no observable phenotypic impact. The cause of this is often attributed to robustness or redundancy within the genome, but that is only one plausible hypothesis. We examined a standard set of phenotypic traits, and applied statistical methods commonly used in the study of natural variants to an engineered mutant strain collection representing disruptions in 180 of the 192 ABC transporters within the bacterium Myxococcus xanthus. These strains display continuous variation in their phenotypic distributions, with a small number of “outlier” strains at both phenotypic extremes, and the majority within a confidence interval about the mean that always includes wild type. Correlation analysis reveals substantial pleiotropy, indicating that the traits do not represent independent variables. The traits measured in this study co-cluster with expression profiles, thereby demonstrating that these changes in phenotype correspond to changes at the molecular level, and therefore can be indirectly connected to changes in the genome. However, the continuous distributions, the pleiotropy, and the placement of wild type always within the confidence interval all indicate that this standard set of M. xanthus phenotypic assays is measuring a narrow range of partially overlapping traits that do not directly reflect fitness. This is likely a significant cause of the observed small phenotypic impact from mutation, and is unrelated to robustness and redundancy.

  3. An asymmetric post-hydrolysis state of the ABC transporter ATPase dimer.

    George, Anthony M; Jones, Peter M

    2013-01-01

    ABC transporters are a superfamily of enzyme pumps that hydrolyse ATP in exchange for translocation of substrates across cellular membranes. Architecturally, ABC transporters are a dimer of transmembrane domains coupled to a dimer of nucleotide binding domains (NBDs): the NBD dimer contains two ATP-binding sites at the intersubunit interface. A current controversy is whether the protomers of the NBD dimer separate during ATP hydrolysis cycling, or remain in constant contact. In order to investigate the ABC ATPase catalytic mechanism, MD simulations using the recent structure of the ADP+Pi-bound MJ0796 isolated NBD dimer were performed. In three independent simulations of the ADP+Pi/apo state, comprising a total of >0.5 µs, significant opening of the apo (empty) active site was observed; occurring by way of intrasubunit rotations between the core and helical subdomains within both NBD monomers. In contrast, in three equivalent simulations of the ATP/apo state, the NBD dimer remained close to the crystal structure, and no opening of either active site occurred. The results thus showed allosteric coupling between the active sites, mediated by intrasubunit conformational changes. Opening of the apo site is exquisitely tuned to the nature of the ligand, and thus to the stage of the reaction cycle, in the opposite site. In addition to this, in also showing how one active site can open, sufficient to bind nucleotide, while the opposite site remains occluded and bound to the hydrolysis products ADP+Pi, the results are consistent with a Constant Contact Model. Conversely, they show how there may be no requirement for the NBD protomers to separate to complete the catalytic cycle. PMID:23573213

  4. An asymmetric post-hydrolysis state of the ABC transporter ATPase dimer.

    Anthony M George

    Full Text Available ABC transporters are a superfamily of enzyme pumps that hydrolyse ATP in exchange for translocation of substrates across cellular membranes. Architecturally, ABC transporters are a dimer of transmembrane domains coupled to a dimer of nucleotide binding domains (NBDs: the NBD dimer contains two ATP-binding sites at the intersubunit interface. A current controversy is whether the protomers of the NBD dimer separate during ATP hydrolysis cycling, or remain in constant contact. In order to investigate the ABC ATPase catalytic mechanism, MD simulations using the recent structure of the ADP+Pi-bound MJ0796 isolated NBD dimer were performed. In three independent simulations of the ADP+Pi/apo state, comprising a total of >0.5 µs, significant opening of the apo (empty active site was observed; occurring by way of intrasubunit rotations between the core and helical subdomains within both NBD monomers. In contrast, in three equivalent simulations of the ATP/apo state, the NBD dimer remained close to the crystal structure, and no opening of either active site occurred. The results thus showed allosteric coupling between the active sites, mediated by intrasubunit conformational changes. Opening of the apo site is exquisitely tuned to the nature of the ligand, and thus to the stage of the reaction cycle, in the opposite site. In addition to this, in also showing how one active site can open, sufficient to bind nucleotide, while the opposite site remains occluded and bound to the hydrolysis products ADP+Pi, the results are consistent with a Constant Contact Model. Conversely, they show how there may be no requirement for the NBD protomers to separate to complete the catalytic cycle.

  5. A PhoPQ-Regulated ABC Transporter System Exports Tetracycline in Pseudomonas aeruginosa.

    Chen, Lin; Duan, Kangmin

    2016-05-01

    Pseudomonas aeruginosa is an important human pathogen whose infections are difficult to treat due to its high intrinsic resistance to many antibiotics. Here, we show that the disruption of PA4456, encoding the ATP binding component of a putative ATP-binding cassette (ABC) transporter, increased the bacterium's susceptible to tetracycline and other antibiotics or toxic chemicals. Fluorescence spectroscopy and antibiotic accumulation tests showed that the interruption of the ABC transporter caused increased intracellular accumulation of tetracycline, demonstrating a role of the ABC transporter in tetracycline expulsion. Site-directed mutagenesis proved that the conserved residues of E170 in the Walker B motif and H203 in the H-loop, which are important for ATP hydrolysis, were essential for the function of PA4456. Through a genome-wide search, the PhoPQ two-component system was identified as a regulator of the computationally predicted PA4456-4452 operon that encodes the ABC transporter system. A >5-fold increase of the expression of this operon was observed in the phoQ mutant. The results obtained also show that the expression of the phzA1B1C1D1E1 operon and the production of pyocyanin were significantly higher in the ABC transporter mutant, signifying a connection between the ABC transporter and pyocyanin production. These results indicated that the PhoPQ-regulated ABC transporter is associated with intrinsic resistance to antibiotics and other adverse compounds in P. aeruginosa, probably by extruding them out of the cell. PMID:26953208

  6. ATP-dependent substrate transport by the ABC transporter MsbA is proton-coupled

    Singh, Himansha; Velamakanni, Saroj; Deery, Michael J.; Howard, Julie; Wei, Shen L.; van Veen, Hendrik W.

    2016-01-01

    ATP-binding cassette transporters mediate the transbilayer movement of a vast number of substrates in or out of cells in organisms ranging from bacteria to humans. Current alternating access models for ABC exporters including the multidrug and Lipid A transporter MsbA from Escherichia coli suggest a role for nucleotide as the fundamental source of free energy. These models involve cycling between conformations with inward- and outward-facing substrate-binding sites in response to engagement and hydrolysis of ATP at the nucleotide-binding domains. Here we report that MsbA also utilizes another major energy currency in the cell by coupling substrate transport to a transmembrane electrochemical proton gradient. The dependence of ATP-dependent transport on proton coupling, and the stimulation of MsbA-ATPase by the chemical proton gradient highlight the functional integration of both forms of metabolic energy. These findings introduce ion coupling as a new parameter in the mechanism of this homodimeric ABC transporter. PMID:27499013

  7. ABC transporters as multidrug resistance mechanisms and the development of chemosensitizers for their reversal

    Choi Cheol-Hee

    2005-10-01

    Full Text Available Abstract One of the major problems related with anticancer chemotherapy is resistance against anticancer drugs. The ATP-binding cassette (ABC transporters are a family of transporter proteins that are responsible for drug resistance and a low bioavailability of drugs by pumping a variety of drugs out cells at the expense of ATP hydrolysis. One strategy for reversal of the resistance of tumor cells expressing ABC transporters is combined use of anticancer drugs with chemosensitizers. In this review, the physiological functions and structures of ABC transporters, and the development of chemosensitizers are described focusing on well-known proteins including P-glycoprotein, multidrug resistance associated protein, and breast cancer resistance protein.

  8. Glycolysis inhibition inactivates ABC transporters to restore drug sensitivity in malignant cells.

    Ayako Nakano

    Full Text Available Cancer cells eventually acquire drug resistance largely via the aberrant expression of ATP-binding cassette (ABC transporters, ATP-dependent efflux pumps. Because cancer cells produce ATP mostly through glycolysis, in the present study we explored the effects of inhibiting glycolysis on the ABC transporter function and drug sensitivity of malignant cells. Inhibition of glycolysis by 3-bromopyruvate (3BrPA suppressed ATP production in malignant cells, and restored the retention of daunorubicin or mitoxantrone in ABC transporter-expressing, RPMI8226 (ABCG2, KG-1 (ABCB1 and HepG2 cells (ABCB1 and ABCG2. Interestingly, although side population (SP cells isolated from RPMI8226 cells exhibited higher levels of glycolysis with an increased expression of genes involved in the glycolytic pathway, 3BrPA abolished Hoechst 33342 exclusion in SP cells. 3BrPA also disrupted clonogenic capacity in malignant cell lines including RPMI8226, KG-1, and HepG2. Furthermore, 3BrPA restored cytotoxic effects of daunorubicin and doxorubicin on KG-1 and RPMI8226 cells, and markedly suppressed subcutaneous tumor growth in combination with doxorubicin in RPMI8226-implanted mice. These results collectively suggest that the inhibition of glycolysis is able to overcome drug resistance in ABC transporter-expressing malignant cells through the inactivation of ABC transporters and impairment of SP cells with enhanced glycolysis as well as clonogenic cells.

  9. Evidence that Bacterial ABC-Type Transporter Imports Free EDTA for Metabolism▿

    Zhang, Hua; Herman, Jacob P.; Bolton, Harvey; Zhang, Zhicheng; Clark, Sue; XUN, Luying

    2007-01-01

    EDTA, a common chelating agent, is becoming a major organic pollutant in the form of metal-EDTA complexes in surface waters, partly due to its recalcitrance to biodegradation. Even an EDTA-degrading bacterium, BNC1, does not degrade stable metal-EDTA complexes. In the present study, an ABC-type transporter was identified for possible uptake of EDTA because the transporter genes and the EDTA monooxygenase gene were expressed from a single operon in BNC1. The ABC-type transporter had a periplas...

  10. Inhibition or knockdown of ABC transporters enhances susceptibility of adult and juvenile schistosomes to Praziquantel.

    Ravi S Kasinathan

    2014-10-01

    Full Text Available Parasitic flatworms of the genus Schistosoma cause schistosomiasis, a neglected tropical disease that affects hundreds of millions. Treatment of schistosomiasis depends almost entirely on the drug praziquantel (PZQ. Though essential to treating and controlling schistosomiasis, a major limitation of PZQ is that it is not active against immature mammalian-stage schistosomes. Furthermore, there are reports of field isolates with heritable reductions in PZQ susceptibility, and researchers have selected for PZQ-resistant schistosomes in the laboratory. P-glycoprotein (Pgp; ABCB1 and other ATP binding cassette (ABC transporters remove a wide variety of toxins and xenobiotics from cells, and have been implicated in multidrug resistance (MDR. Changes in ABC transporter structure or expression levels are also associated with reduced drug susceptibility in parasitic helminths, including schistosomes. Here, we show that the activity of PZQ against schistosome adults and juveniles ex vivo is potentiated by co-administration of either the highly potent Pgp inhibitor tariquidar or combinations of inhibitors targeting multiple ABC multidrug transporters. Adult worms exposed to sublethal PZQ concentrations remain active, but co-administration of ABC transporter inhibitors results in complete loss of motility and disruption of the tegument. Notably, juvenile schistosomes (3-4 weeks post infection, normally refractory to 2 µM PZQ, become paralyzed when transporter inhibitors are added in combination with the PZQ. Experiments using the fluorescent PZQ derivative (R-PZQ-BODIPY are consistent with the transporter inhibitors increasing effective intraworm concentrations of PZQ. Adult worms in which expression of ABC transporters has been suppressed by RNA interference show increased responsiveness to PZQ and increased retention of (R-PZQ-BODIPY consistent with an important role for these proteins in setting levels of PZQ susceptibility. These results indicate that

  11. Inhibition of ABC transport proteins by oil sands process affected water.

    Alharbi, Hattan A; Saunders, David M V; Al-Mousa, Ahmed; Alcorn, Jane; Pereira, Alberto S; Martin, Jonathan W; Giesy, John P; Wiseman, Steve B

    2016-01-01

    The ATP-binding cassette (ABC) superfamily of transporter proteins is important for detoxification of xenobiotics. For example, ABC transporters from the multidrug-resistance protein (MRP) subfamily are important for excretion of polycyclic aromatic hydrocarbons (PAHs) and their metabolites. Effects of chemicals in the water soluble organic fraction of relatively fresh oil sands process affected water (OSPW) from Base Mine Lake (BML-OSPW) and aged OSPW from Pond 9 (P9-OSPW) on the activity of MRP transporters were investigated in vivo by use of Japanese medaka at the fry stage of development. Activities of MRPs were monitored by use of the lipophilic dye calcein, which is transported from cells by ABC proteins, including MRPs. To begin to identify chemicals that might inhibit activity of MRPs, BML-OSPW and P9-OSPW were fractionated into acidic, basic, and neutral fractions by use of mixed-mode sorbents. Chemical compositions of fractions were determined by use of ultrahigh resolution orbitrap mass spectrometry in ESI(+) and ESI(-) mode. Greater amounts of calcein were retained in fry exposed to BML-OSPW at concentration equivalents greater than 1× (i.e., full strength). The neutral and basic fractions of BML-OSPW, but not the acidic fraction, caused greater retention of calcein. Exposure to P9-OSPW did not affect the amount of calcein in fry. Neutral and basic fractions of BML-OSPW contained relatively greater amounts of several oxygen-, sulfur, and nitrogen-containing chemical species that might inhibit MRPs, such as O(+), SO(+), and NO(+) chemical species, although secondary fractionation will be required to conclusively identify the most potent inhibitors. Naphthenic acids (O2(-)), which were dominant in the acidic fraction, did not appear to be the cause of the inhibition. This is the first study to demonstrate that chemicals in the water soluble organic fraction of OSPW inhibit activity of this important class of proteins. However, aging of OSPW attenuates

  12. Overcoming ABC transporter-mediated multidrug resistance: Molecular mechanisms and novel therapeutic drug strategies.

    Li, Wen; Zhang, Han; Assaraf, Yehuda G; Zhao, Kun; Xu, Xiaojun; Xie, Jinbing; Yang, Dong-Hua; Chen, Zhe-Sheng

    2016-07-01

    Multidrug resistance is a key determinant of cancer chemotherapy failure. One of the major causes of multidrug resistance is the enhanced efflux of drugs by membrane ABC transporters. Targeting ABC transporters projects a promising approach to eliminating or suppressing drug resistance in cancer treatment. To reveal the functional mechanisms of ABC transporters in drug resistance, extensive studies have been conducted from identifying drug binding sites to elucidating structural dynamics. In this review article, we examined the recent crystal structures of ABC proteins to depict the functionally important structural elements, such as domains, conserved motifs, and critical amino acids that are involved in ATP-binding and drug efflux. We inspected the drug-binding sites on ABC proteins and the molecular mechanisms of various substrate interactions with the drug binding pocket. While our continuous battle against drug resistance is far from over, new approaches and technologies have emerged to push forward our frontier. Most recent developments in anti-MDR strategies include P-gp inhibitors, RNA-interference, nano-medicines, and delivering combination strategies. With the advent of the 'Omics' era - genomics, epigenomics, transcriptomics, proteomics, and metabolomics - these disciplines play an important role in fighting the battle against chemoresistance by further unraveling the molecular mechanisms of drug resistance and shed light on medical therapies that specifically target MDR. PMID:27449595

  13. Surface charge-specific interactions between polymer nanoparticles and ABC transporters in Caco-2 cells

    Bhattacharjee, Sourav, E-mail: sourav.bhattacharjee@wur.nl [Wageningen University, Laboratory of Organic Chemistry (Netherlands); Opstal, Edward J. van; Alink, Gerrit M. [Wageningen University, Division of Toxicology (Netherlands); Marcelis, Antonius T. M.; Zuilhof, Han [Wageningen University, Laboratory of Organic Chemistry (Netherlands); Rietjens, Ivonne M. C. M. [Wageningen University, Division of Toxicology (Netherlands)

    2013-06-15

    The surface charge-dependent transport of polymeric nanoparticles (PNPs) across Caco-2 monolayers grown on transwell culture systems as an in vitro model for intestinal transport was tested. The transport of well-characterized, monodisperse, and fluorescent tri-block copolymer nanoparticles (TCNPs/size {approx}45 nm) and polystyrene nanoparticles (PSNPs/size {approx}50 nm), with different surface charges (positive and negative), was quantified. The positive PNPs showed a higher intracellular uptake and flux across the Caco-2 monolayers than the negative PNPs. Multidrug resistance/P-glycoprotein (MDR1/P-gp), a specific ATP-binding cassette (ABC) transporter, was found to play a major role in the cellular efflux of positive PNPs, whereas the multidrug resistance protein 1 took part in the efflux of negative PNPs from Caco-2 cells. The positive PNPs also caused an increased cellular uptake and apical to basolateral transport of the carcinogen PhIP across the Caco-2 monolayer. The flavonoid quercetin, which is known to interact with ABC transporters, promoted the intracellular uptake of different PNPs and interfered with the normal distribution patterns of PNPs in the transwell system. These results indicate that PNPs display surface charge-specific interactions with ABC transporters and can even affect the bioavailability of toxic food-borne compounds (like pro-carcinogens).

  14. Sensitive and specific fluorescent probes for functional analysis of the three major types of mammalian ABC transporters.

    Irina V Lebedeva

    Full Text Available An underlying mechanism for multi drug resistance (MDR is up-regulation of the transmembrane ATP-binding cassette (ABC transporter proteins. ABC transporters also determine the general fate and effect of pharmaceutical agents in the body. The three major types of ABC transporters are MDR1 (P-gp, P-glycoprotein, ABCB1, MRP1/2 (ABCC1/2 and BCRP/MXR (ABCG2 proteins. Flow cytometry (FCM allows determination of the functional expression levels of ABC transporters in live cells, but most dyes used as indicators (rhodamine 123, DiOC(2(3, calcein-AM have limited applicability as they do not detect all three major types of ABC transporters. Dyes with broad coverage (such as doxorubicin, daunorubicin and mitoxantrone lack sensitivity due to overall dimness and thus may yield a significant percentage of false negative results. We describe two novel fluorescent probes that are substrates for all three common types of ABC transporters and can serve as indicators of MDR in flow cytometry assays using live cells. The probes exhibit fast internalization, favorable uptake/efflux kinetics and high sensitivity of MDR detection, as established by multidrug resistance activity factor (MAF values and Kolmogorov-Smirnov statistical analysis. Used in combination with general or specific inhibitors of ABC transporters, both dyes readily identify functional efflux and are capable of detecting small levels of efflux as well as defining the type of multidrug resistance. The assay can be applied to the screening of putative modulators of ABC transporters, facilitating rapid, reproducible, specific and relatively simple functional detection of ABC transporter activity, and ready implementation on widely available instruments.

  15. Transportation System Requirements Document

    1993-09-01

    This Transportation System Requirements Document (Trans-SRD) describes the functions to be performed by and the technical requirements for the Transportation System to transport spent nuclear fuel (SNF) and high-level radioactive waste (HLW) from Purchaser and Producer sites to a Civilian Radioactive Waste Management System (CRWMS) site, and between CRWMS sites. The purpose of this document is to define the system-level requirements for Transportation consistent with the CRWMS Requirement Document (CRD). These requirements include design and operations requirements to the extent they impact on the development of the physical segments of Transportation. The document also presents an overall description of Transportation, its functions, its segments, and the requirements allocated to the segments and the system-level interfaces with Transportation. The interface identification and description are published in the CRWMS Interface Specification.

  16. Transportation System Requirements Document

    This Transportation System Requirements Document (Trans-SRD) describes the functions to be performed by and the technical requirements for the Transportation System to transport spent nuclear fuel (SNF) and high-level radioactive waste (HLW) from Purchaser and Producer sites to a Civilian Radioactive Waste Management System (CRWMS) site, and between CRWMS sites. The purpose of this document is to define the system-level requirements for Transportation consistent with the CRWMS Requirement Document (CRD). These requirements include design and operations requirements to the extent they impact on the development of the physical segments of Transportation. The document also presents an overall description of Transportation, its functions, its segments, and the requirements allocated to the segments and the system-level interfaces with Transportation. The interface identification and description are published in the CRWMS Interface Specification

  17. Effects of the L511P and D512G Mutations on the Escherichia coli ABC Transporter MsbA

    Schultz, Kathryn M.; Merten, Jacqueline A.; Klug, Candice S.

    2011-01-01

    MsbA is a member of the ABC transporter superfamily and is homologous to ABC transporters linked to multidrug resistance. The nucleotide binding domains (NBDs) of these proteins include conserved motifs that are involved in ATP binding, including conserved SALD residues (D-loop) that are diagnostic in identifying ABC transporters but whose roles have not been identified. Within the D-loop, single point mutations L511P and D512G were discovered by random mutational analysis of MsbA to disrupt ...

  18. ABC transporters involved in the biogenesis of the outer membrane in gram-negative bacteria.

    Narita, Shin-ichiro

    2011-01-01

    The outer membrane of gram-negative bacteria is an asymmetric lipid bilayer with phospholipids and lipopolysaccharides (LPSs). β-Barreled outer membrane proteins and lipoproteins are embedded in the outer membrane. All of these constituents are essential to the function of the outer membrane. The transport systems for lipoproteins have been characterized in detail. An ATP-binding cassette (ABC) transporter, LolCDE, initiates sorting by mediating the detachment of lipoproteins from the inner membrane to form a water-soluble lipoprotein-LolA complex in the periplasm. Lipoproteins are then transferred to LolB at the outer membrane and are incorporated into the lipid bilayer. A model analogous to the Lol system has been suggested for the transport of LPS, where an ABC transporter, LptBFG, mediates the detachment of LPS from the inner membrane. Recent developments in the functional characterization of ABC transporters involved in the biogenesis of the outer membrane in gram-negative bacteria are discussed. PMID:21670534

  19. Functional analysis of ABC transporter genes from Botrytis cinerea identifies BcatrB as a transporter of eugenol

    Schoonbeek, H.; Nistelrooy, van J.G.M.; Waard, de M.A.

    2003-01-01

    The role of multiple ATP-binding cassette (ABC) and major facilitator superfamily (MFS) transporter genes from the plant pathogenic fungus Botrytis cinerea in protection against natural fungitoxic compounds was studied by expression analysis and phenotyping of gene-replacement mutants. The expressio

  20. Nucleotide-induced conformational dynamics in ABC transporters from structure-based coarse grained modelling.

    Flechsig, Holger

    2016-02-01

    ATP-binding cassette (ABC) transporters are integral membrane proteins which mediate the exchange of diverse substrates across membranes powered by ATP molecules. Our understanding of their activity is still hampered since the conformational dynamics underlying the operation of such proteins cannot yet be resolved in detailed molecular dynamics studies. Here a coarse grained model which allows to mimic binding of nucleotides and follow subsequent conformational motions of full-length transporter structures in computer simulations is proposed and implemented. To justify its explanatory quality, the model is first applied to the maltose transporter system for which multiple conformations are known and we find that the model predictions agree remarkably well with the experimental data. For the MalK subunit the switching from open to the closed dimer configuration upon ATP binding is reproduced and, moreover, for the full-length maltose transporter, progression from inward-facing to the outward-facing state is correctly obtained. For the heme transporter HmuUV, for which only the free structure could yet be determined, the model was then applied to predict nucleotide-induced conformational motions. Upon binding of ATP-mimicking ligands the structure changed from a conformation in which the nucleotide-binding domains formed an open shape, to a conformation in which they were found in tight contact, while, at the same time, a pronounced rotation of the transmembrane domains was observed. This finding is supported by normal mode analysis, and, comparison with structural data of the homologous vitamin B12 transporter BtuCD suggests that the observed rotation mechanism may contribute a common functional aspect for this class of ABC transporters. Although in HmuuV noticeable rearrangement of essential transmembrane helices was detected, there are no indications from our simulations that ATP binding alone may facilitate propagation of substrate molecules in this transporter

  1. ABC transporters and xenobiotic defense systems in early life stages of rainbow trout (Oncorhynchus mykiss).

    Kropf, Christian; Segner, Helmut; Fent, Karl

    2016-01-01

    Embryos of oviparous fish, in contrast to (ovo) viviparous species, develop in the aquatic environment, and therefore need solute transport systems at their body surfaces for maintaining internal homeostasis and defending against potentially harmful substances. We hypothesized that solute transporters undergo changes in tissue distribution from the embryo to the larval stage. We therefore studied the mRNA profiles of eight ABC transporters (abcb1a, abcb1b, abcc1, abcc2, abcc3, abcc4, abcc5, abcg2) and three solute carriers (oatp1d, putative oatp2 putative, mate1) in different body regions (head, yolk sac epithelium, abdominal viscera, skin/muscles) of developing rainbow trout. Additionally, we investigated mRNA levels of phase I (cyp1a, cyp3a) and phase II (gstp, putative ugt1, putative ugt2) biotransformation enzymes. The study covered the developmental period from the eleuthero-embryo stage to the first-feeding larval stage (1-20days post-hatch, dph). At 1dph, transcripts of abcc2, abcc4, abcg2, cyp3a, gstp, putative mate1, and putative oatp2 occurred primarily in the yolk sac epithelium, whereas at later stages expression of these genes was predominantly observed in the abdominal viscera. The functional activity of ABC transporters in fish early life stages was assessed by rhodamine B accumulation assays. Finally, we investigated the potential impact of xenobiotics (clotrimazole, clofibric acid) on the ABC and biotransformation systems of trout early life stages. While clofibric acid had no effect, clotrimazole lead to an increased rhodamine B accumulation. The results provide evidence that the transition from the eleuthero-embryo to the larval stage is accompanied by a major alteration in tissue expression of ABC transporters. PMID:26945521

  2. Functionally Relevant Residues of Cdr1p: A Multidrug ABC Transporter of Human Pathogenic Candida albicans

    Rajendra Prasad

    2011-01-01

    Full Text Available Reduced intracellular accumulation of drugs (due to rapid efflux mediated by the efflux pump proteins belonging to ABC (ATP Binding Cassette and MFS (Major Facilitators superfamily is one of the most common strategies adopted by multidrug resistance (MDR pathogenic yeasts. To combat MDR, it is essential to understand the structure and function of these transporters so that inhibitors/modulators to these can be developed. The sequence alignments of the ABC transporters reveal selective divergence within much conserved domains of Nucleotide-Binding Domains (NBDs which is unique to all fungal transporters. Recently, the role of conserved but divergent residues of Candida Drug Resistance 1 (CDR1, an ABC drug transporter of human pathogenic Candida albicans, has been examined with regard to ATP binding and hydrolysis. In this paper, we focus on some of the recent advances on the relevance of divergent and conserved amino acids of CaCdr1p and also discuss as to how drug interacts with Trans Membrane Domains (TMDs residues for its extrusion from MDR cells.

  3. Direct Observation of a Gate Tunable Band Gap in Electrical Transport in ABC-Trilayer Graphene.

    Khodkov, Tymofiy; Khrapach, Ivan; Craciun, Monica Felicia; Russo, Saverio

    2015-07-01

    Few layer graphene systems such as Bernal stacked bilayer and rhombohedral (ABC-) stacked trilayer offer the unique possibility to open an electric field tunable energy gap. To date, this energy gap has been experimentally confirmed in optical spectroscopy. Here we report the first direct observation of the electric field tunable energy gap in electronic transport experiments on doubly gated suspended ABC-trilayer graphene. From a systematic study of the nonlinearities in current versus voltage characteristics and the temperature dependence of the conductivity, we demonstrate that thermally activated transport over the energy-gap dominates the electrical response of these transistors. The estimated values for energy gap from the temperature dependence and from the current voltage characteristics follow the theoretically expected electric field dependence with critical exponent 3/2. These experiments indicate that high quality few-layer graphene are suitable candidates for exploring novel tunable terahertz light sources and detectors. PMID:26079989

  4. A putative ABC transporter is involved in negative regulation of biofilm formation by Listeria monocytogenes

    Zhu, Xinna; Long, Fei; Chen, Yonghui;

    2008-01-01

    Listeria monocytogenes may persist for long periods in food processing environments. In some instances, this may be due to aggregation or biofilm formation. To investigate the mechanism controlling biofilm formation in the food-borne pathogen L. monocytogenes, we characterized LM-49, a mutant with...... enhanced ability of biofilm-formation generated via transposon Tn917 mutagenesis of L. monocytogenes 4b G. In this mutant, a Tn917 insertion has disrupted the coding region of the gene encoding a putative ATP binding cassette (ABC) transporter permease identical to Lmof2365_1771 (a putative ABC......-transporter permease) presented in the sequenced strain L. monocytogenes str. 4b F2365. This disrupted gene, denoted lm.G_1771, encoded a protein with 10 transmembrane helixes. The revertant, LM-49RE, was obtained by replacing lm.G_1771::Tn917 with lm.G_1771 via homologous recombination. We found that LM-49RE formed...

  5. ABC transporters in CSCs membranes as a novel target for treating tumour relapse

    LAURA eZINZI; Marialessandra eContino; Mariangela eCantore; Elena eCapparelli; Marcello eLeopoldo; Nicola Antonio Colabufo

    2014-01-01

    CSCs are responsible for the high rate of recurrence and chemoresistance of different type of cancers. The current antineoplastic agents, able to inhibit bulk replicating cancer cells and radiation treatment, were found inefficacious towards CSCs since this subpopulation has several intrinsic mechanisms of resistance. Among these mechanisms, the expression of ATP-Binding Cassette (ABC) transporters family and the activation of different signaling pathway (such as Wnt/β-catenin signaling, Hedg...

  6. ABC transporters in CSCs membranes as a novel target for treating tumor relapse

    Zinzi, Laura; Contino, Marialessandra; Cantore, Mariangela; Capparelli, Elena; Leopoldo, Marcello; Nicola A. Colabufo

    2014-01-01

    CSCs are responsible for the high rate of recurrence and chemoresistance of different types of cancer. The current antineoplastic agents able to inhibit bulk replicating cancer cells and radiation treatment are not efficacious toward CSCs since this subpopulation has several intrinsic mechanisms of resistance. Among these mechanisms, the expression of ATP-Binding Cassette (ABC) transporters family and the activation of different signaling pathways (such as Wnt/β-catenin signaling, Hedgehog, N...

  7. Deoxycytidine kinase modulates the impact of the ABC transporter ABCG2 on clofarabine cytotoxicity

    Nagai, Shinjiro; Takenaka, Kazumasa; Nachagari, Deepa; Rose, Charles; Domoney, Kali; Sun, Daxi; Sparreboom, Alex; Schuetz, John D.

    2011-01-01

    Purine nucleoside antimetabolites, such as clofarabine, are effective antileukemic agents. However, their effectiveness depends on an initial activation step in which they are monophosphorylated by deoxycytidine kinase (dCK). Some purine nucleoside antimetabolites and their monophosphate derivatives are exported by the ABC transporter ABCG2. Because clofarabine is a dCK substrate, and we show substantial variation in dCK and ABCG2 in myeloid leukemia, we hypothesized that the activity of dCK ...

  8. CD4+ T cell immunity to the Burkholderia pseudomallei ABC transporter LolC in melioidosis

    Chu, Karen K.; Tippayawat, Patcharaporn; Walker, Nicola J.; Harding, Sarah V.; Atkins, Helen S.; Maillere, Bernard; Bancroft, Gregory J; Lertmemongkolchai, Ganjana; Altmann, Daniel M

    2010-01-01

    Burkholderia pseudomallei (Bp) causes melioidosis, a disease with a wide range of possible outcomes, from seroconversion and dormancy to sepsis and death. This spectrum of host-pathogen interactions poses challenging questions about heterogeneity in immunity to Bp. Models show protection to be dependent on CD4+ cells and IFNγ, but little is known about specific target antigens. Having previously implicated the ABC transporter, LolC, in protective immunity, we here use epitope prediction, HLA ...

  9. Effect of selected ABC-drug transporters and anticancer drug disposition in vitro and in vivo

    Marchetti, S

    2013-01-01

    Studies described in the thesis that is lying in front of you aim to address the possible implications of selected ABC-drug transporters on the disposition of a number of important anticancer drugs. Although variability in drug disposition has been known for as long as pharmacological studies supported drug development and clinical therapeutics general molecular pharmacological concepts explaining the given interpatient variation in drug disposition have been lacking for many decades. Firm ex...

  10. RLIP76, a non-ABC transporter, and drug resistance in epilepsy

    Awasthi Yogesh C; Cucullo Luca; Singhal Sharad S; Fazio Vince; Hallene Kerri L; Awasthi Sanjay; Dini Gabriele; Janigro Damir

    2005-01-01

    Abstract Background Permeability of the blood-brain barrier is one of the factors determining the bioavailability of therapeutic drugs and resistance to chemically different antiepileptic drugs is a consequence of decreased intracerebral accumulation. The ABC transporters, particularly P-glycoprotein, are known to play a role in antiepileptic drug extrusion, but are not by themselves sufficient to fully explain the phenomenon of drug-resistant epilepsy. Proteomic analyses of membrane protein ...

  11. Structural arrangement of the transmission interface in the antigen ABC transport complex TAP

    Oancea, Giani; O'Mara, Megan L.; Bennett, W.F. Drew; Tieleman, D. Peter; Abele, Rupert; Tampé, Robert

    2009-01-01

    The transporter associated with antigen processing (TAP) represents a focal point in the immune recognition of virally or malignantly transformed cells by translocating proteasomal degradation products into the endoplasmic reticulum–lumen for loading of MHC class I molecules. Based on a number of experimental data and the homology to the bacterial ABC exporter Sav1866, we constructed a 3D structural model of the core TAP complex and used it to examine the interface between the transmembrane a...

  12. ABC transporters in CSCs membranes as a novel target for treating tumour relapse

    LAURA eZINZI

    2014-07-01

    Full Text Available CSCs are responsible for the high rate of recurrence and chemoresistance of different type of cancers. The current antineoplastic agents, able to inhibit bulk replicating cancer cells and radiation treatment, were found inefficacious towards CSCs since this subpopulation has several intrinsic mechanisms of resistance. Among these mechanisms, the expression of ATP-Binding Cassette (ABC transporters family and the activation of different signaling pathway (such as Wnt/β-catenin signaling, Hedgehog, Notch, Akt/PKB are reported. Therefore, considering ABC transporters expression on CSCs membranes, compounds able to modulate MDR could induce cytotoxicity in these cells disclosing an exciting and alternative strategy for targeting CSCs in tumour therapy. The next challenge in the cure of cancer relapse may be a multimodal strategy, an approach in which specific CSCs targeting drugs exert simultaneously the ability to circumvent tumor drug resistance (ABC transporters modulation and cytotoxic activity towards CSCs and the corresponding differentiated tumour cells. The efficacy of suggested multimodal strategy could be probed by using several scaffolds active towards MDR pumps on CSCs isolated by tumour specimens.

  13. The ABC family of multidrug transporters in microorganisms

    van Veen, H.W; Konings, W.N

    1998-01-01

    Multidrug transporters are membrane proteins that are able to expel a broad range of toxic molecules from the cell. In humans, the overexpression of the multidrug resistance P-glycoprotein (Pgp) and the multidrug resistance-associated protein MRP1 (MRP) is a principal cause of resistance of cancers

  14. [Role of ABC efflux transporters in the oral bioavailability and drug-induced intestinal toxicity].

    Yokooji, Tomoharu

    2013-01-01

    The gastrointestinal tract is the organ that absorbs nutrients and water from foods and drinks. This organ is often exposed to various harmful xenobiotics, and therefore possesses various detoxification/barrier systems, including metabolizing enzymes and efflux transporters. Intestinal epithelial cells express ATP-binding cassette (ABC) efflux transporters such as P-glycoprotein, multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein, in addition to various solute carrier (SLC) influx transporters. These transporters are expressed site- and membrane-specifically in enterocytes, which affects the bioavailability of ingested substrate drugs. Expression and/or function of transporters can be modulated by various compounds, including therapeutic drugs, herbal products, some foods, and by disease states. The modulation of transporters could cause unexpectedly higher or lower blood concentrations, marked inter- and intra-individual variations in pharmacokinetics, and unreliable pharmacological actions in association with toxicities of substrates. Recently, we found that hyperbilirubinemia, which occurs in some disease states, increased intestinal accumulation and toxicity of methotrexate, an MRP substrate, because of the suppression of MRP function by high plasma concentrations of conjugated bilirubin. We also attempted to ameliorate the intestinal toxicity of irinotecan hydrochloride by modulating the hepatic and intestinal functions of MRP2. This review summarizes our findings regarding the role of ABC transporters, especially MRPs, in oral bioavailability and in drug-induced intestinal toxicity. Our approach to treat intestinal toxicity using an MRP2 modulator is also described. PMID:23811769

  15. ATP-Binding Cassette (ABC) Transporters of the Human Respiratory Tract Pathogen, Moraxella catarrhalis: Role in Virulence.

    Murphy, Timothy F; Brauer, Aimee L; Johnson, Antoinette; Kirkham, Charmaine

    2016-01-01

    Moraxella catarrhalis is a human respiratory tract pathogen that causes otitis media (middle ear infections) in children and respiratory tract infections in adults with chronic obstructive pulmonary disease. In view of the huge global burden of disease caused by M. catarrhalis, the development of vaccines to prevent these infections and better approaches to treatment have become priorities. In previous work, we used a genome mining approach that identified three substrate binding proteins (SBPs) of ATP-binding cassette (ABC) transporters as promising candidate vaccine antigens. In the present study, we performed a comprehensive assessment of 19 SBPs of 15 ABC transporter systems in the M. catarrhalis genome by engineering knockout mutants and studying their role in assays that assess mechanisms of infection. The capacity of M. catarrhalis to survive and grow in the nutrient-limited and hostile environment of the human respiratory tract, including intracellular growth, account in part for its virulence. The results show that ABC transporters that mediate uptake of peptides, amino acids, cations and anions play important roles in pathogenesis by enabling M. catarrhalis to 1) grow in nutrient-limited conditions, 2) invade and survive in human respiratory epithelial cells and 3) persist in the lungs in a murine pulmonary clearance model. The knockout mutants of SBPs and ABC transporters showed different patterns of activity in the assay systems, supporting the conclusion that different SBPs and ABC transporters function at different stages in the pathogenesis of infection. These results indicate that ABC transporters are nutritional virulence factors, functioning to enable the survival of M catarrhalis in the diverse microenvironments of the respiratory tract. Based on the role of ABC transporters as virulence factors of M. catarrhalis, these molecules represent potential drug targets to eradicate the organism from the human respiratory tract. PMID:27391026

  16. Pharmacogenomics of the human ABC transporter ABCG2: from functional evaluation to drug molecular design

    Ishikawa, Toshihisa; Tamura, Ai; Saito, Hikaru; Wakabayashi, Kanako; Nakagawa, Hiroshi

    2005-10-01

    In the post-genome-sequencing era, emerging genomic technologies are shifting the paradigm for drug discovery and development. Nevertheless, drug discovery and development still remain high-risk and high-stakes ventures with long and costly timelines. Indeed, the attrition of drug candidates in preclinical and development stages is a major problem in drug design. For at least 30% of the candidates, this attrition is due to poor pharmacokinetics and toxicity. Thus, pharmaceutical companies have begun to seriously re-evaluate their current strategies of drug discovery and development. In that light, we propose that a transport mechanism-based design might help to create new, pharmacokinetically advantageous drugs, and as such should be considered an important component of drug design strategy. Performing enzyme- and/or cell-based drug transporter, interaction tests may greatly facilitate drug development and allow the prediction of drug-drug interactions. We recently developed methods for high-speed functional screening and quantitative structure-activity relationship analysis to study the substrate specificity of ABC transporters and to evaluate the effect of genetic polymorphisms on their function. These methods would provide a practical tool to screen synthetic and natural compounds, and these data can be applied to the molecular design of new drugs. In this review article, we present an overview on the genetic polymorphisms of human ABC transporter ABCG2 and new camptothecin analogues that can circumvent AGCG2-associated multidrug resistance of cancer.

  17. The Heterodimeric ABC Transporter EfrCD Mediates Multidrug Efflux in Enterococcus faecalis.

    Hürlimann, Lea M; Corradi, Valentina; Hohl, Michael; Bloemberg, Guido V; Tieleman, D Peter; Seeger, Markus A

    2016-09-01

    Nosocomial infections with Enterococcus faecalis are an emerging health problem. However, drug efflux pumps contributing to intrinsic drug resistance are poorly studied in this Gram-positive pathogen. In this study, we functionally investigated seven heterodimeric ABC transporters of E. faecalis that are annotated as drug efflux pumps. Deletion of ef0789-ef0790 on the chromosome of E. faecalis resulted in increased susceptibility to daunorubicin, doxorubicin, ethidium, and Hoechst 33342, and the corresponding transporter was named EfrCD. Unexpectedly, the previously described heterodimeric multidrug ABC transporter EfrAB contributes marginally to drug efflux in the endogenous context of E. faecalis In contrast, heterologous expression in Lactococcus lactis revealed that EfrAB, EfrCD, and the product of ef2226-ef2227 (EfrEF) mediate the efflux of fluorescent substrates and confer resistance to multiple dyes and drugs, including fluoroquinolones. Four of seven transporters failed to exhibit drug efflux activity for the set of drugs and dyes tested, even upon overexpression in L. lactis Since all seven transporters were purified as heterodimers after overexpression in L. lactis, a lack of drug efflux activity is not attributed to poor expression or protein aggregation. Reconstitution of the purified multidrug transporters EfrAB, EfrCD, and EfrEF in proteoliposomes revealed functional coupling between ATP hydrolysis and drug binding. Our analysis creates an experimental basis for the accurate prediction of drug efflux transporters and indicates that many annotated multidrug efflux pumps might be incapable of drug transport and thus might fulfill other physiological functions in the cell. PMID:27381387

  18. Gene expression of ABC transporters in Cooperia oncophora after field and laboratory selection with macrocyclic lactones.

    Tydén, Eva; Skarin, Moa; Höglund, Johan

    2014-12-01

    The most widespread helminth parasites of grazing cattle in northern Europe are the gastrointestinal nematodes Ostertagia ostertagi and Cooperia oncophora. Heavy reliance on the use of macrocyclic lactone (ML) in cattle has led to world-wide emergence of resistance to this drug class in C. oncophora. There is evidence that members of the ATP-binding cassette (ABC) transporter family, such as P-glycoproteins (P-gp) and multidrug-resistant proteins (MRP), play a role in resistance to ML. In this study gene expression of Con-pgp9, Con-pgp11, Con-pgp12, Con-pgp16 and Con-mrp1 was examined in two isolates of C. oncophora sharing the same genetic background but exposed to ML differently. For isolate one (Laboratory-selected), adult worms were recovered before and after treatment with ML in vivo. For isolate two (Field-selected), adult worms were collected from tracer animals that had never received anthelmintics themselves. One group grazed together with untreated animals and one group grazed with animals that received suppressive prophylactic treatment with ML at monthly intervals for up to two consecutive grazing seasons. Real-time PCR data demonstrated differences in gene expression after ML selection, with the highest constitutive expression levels for Con-pgp16 and Con-mrp1. Remarkably, the same pattern of increasing expression levels of the ABC transport genes was observed in both Laboratory- and Field-selected isolates, despite the Field-selected isolate not being directly exposed to ML. The higher expression levels of ABC transporters observed in the Field-selected isolate was thus not a response to direct exposure to ML, but rather appeared to reflect a genetic characteristic inherited from worms in the previous generation which had survived exposure to ML in the co-grazing treated animals. PMID:25619799

  19. Distribution and Genetic Diversity of the ABC Transporter Lipoproteins PiuA and PiaA within Streptococcus pneumoniae and Related Streptococci

    Whalan, Rachael H.; Funnell, Simon G.P.; Bowler, Lucas D.; Hudson, Michael J.; Robinson, Andrew; Dowson, Christopher G.

    2006-01-01

    Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. The existence of approximately 90 antigenically distinct capsular serotypes has greatly complicated the development of an effective pneumococcal vaccine. Virulence-associated proteins common and conserved among all capsular types now represent the best strategy to combat pneumococcal infections. PiuA and PiaA are the lipoprotein components of two pneumococcal iron ABC transporters and are required for full virulen...

  20. Integrated analysis of residue coevolution and protein structure in ABC transporters.

    Attila Gulyás-Kovács

    Full Text Available Intraprotein side chain contacts can couple the evolutionary process of amino acid substitution at one position to that at another. This coupling, known as residue coevolution, may vary in strength. Conserved contacts thus not only define 3-dimensional protein structure, but also indicate which residue-residue interactions are crucial to a protein's function. Therefore, prediction of strongly coevolving residue-pairs helps clarify molecular mechanisms underlying function. Previously, various coevolution detectors have been employed separately to predict these pairs purely from multiple sequence alignments, while disregarding available structural information. This study introduces an integrative framework that improves the accuracy of such predictions, relative to previous approaches, by combining multiple coevolution detectors and incorporating structural contact information. This framework is applied to the ABC-B and ABC-C transporter families, which include the drug exporter P-glycoprotein involved in multidrug resistance of cancer cells, as well as the CFTR chloride channel linked to cystic fibrosis disease. The predicted coevolving pairs are further analyzed based on conformational changes inferred from outward- and inward-facing transporter structures. The analysis suggests that some pairs coevolved to directly regulate conformational changes of the alternating-access transport mechanism, while others to stabilize rigid-body-like components of the protein structure. Moreover, some identified pairs correspond to residues previously implicated in cystic fibrosis.

  1. Lysimachia foenum-graecum Herba Extract, a Novel Biopesticide, Inhibits ABC Transporter Genes and Mycelial Growth of Magnaporthe oryzae.

    Lee, Youngjin

    2016-02-01

    To identify a novel biopesticide controlling rice blast disease caused by Magnaporthe oryzae, 700 plant extracts were evaluated for their inhibitory effects on mycelial growth of M. oryzae. The L. foenum-graecum Herba extract showed the lowest inhibition concentration (IC50) of 39.28 μg/ml, which is lower than the IC50 of blasticidin S (63.06 μg/ml), a conventional fungicide for rice blast disease. When treatments were combined, the IC50 of blasticidin S was dramatically reduced to 10.67 μg/ml. Since ABC transporter genes are involved in fungicide resistance of many organisms, we performed RT-PCR to investigate the transcriptional changes of 40 ABC transporter family genes of M. oryzae treated with the plant extract, blasticidin S, and tetrandrine, a recognized ABC transporter inhibitor. Four ABC transporter genes were prominently activated by blasticidin S treatment, but were suppressed by combinational treatment of blasticidin S with the plant extract, or with tetrandrine that didn't show cellular toxicity by itself in this study. Mycelial death was detected via confocal microscopy at 24 h after plant extract treatment. Finally, subsequent rice field study revealed that the plant extract had high control efficacy of 63.3% and should be considered a biopesticide for rice blast disease. These results showed that extract of L. foenum graecum Herba suppresses M. oryzae ABC transporter genes inducing mycelial death and therefore may be a potent novel biopesticide. PMID:26889110

  2. A single intact ATPase site of the ABC transporter BtuCD drives 5% transport activity yet supports full in vivo vitamin B12 utilization

    Tal, Nir; Ovcharenko, Elena; Lewinson, Oded

    2013-01-01

    In all kingdoms of life, ATP binding cassette (ABC) transporters are essential to many cellular functions. In this large superfamily of proteins, two catalytic sites hydrolyze ATP to power uphill substrate translocation. A central question in the field concerns the relationship between the two ATPase catalytic sites: Are the sites independent of one another? Are both needed for function? Do they function cooperatively? These issues have been resolved for type I ABC transporters but never for ...

  3. The Klebsiella pneumoniae O12 ATP-binding Cassette (ABC) Transporter Recognizes the Terminal Residue of Its O-antigen Polysaccharide Substrate.

    Mann, Evan; Mallette, Evan; Clarke, Bradley R; Kimber, Matthew S; Whitfield, Chris

    2016-04-29

    Export of the Escherichia coli serotype O9a O-antigenic polysaccharides (O-PS) involves an ATP-binding cassette (ABC) transporter. The process requires a non-reducing terminal residue, which is recognized by a carbohydrate-binding module (CBM) appended to the C terminus of the nucleotide-binding domain of the transporter. Here, we investigate the process in Klebsiella pneumoniae serotype O12 (and Raoultella terrigena ATCC 33257). The O12 polysaccharide is terminated at the non-reducing end by a β-linked 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) residue. The O12 ABC transporter also binds its cognate O-PS via a CBM, and export is dependent on the presence of the terminal β-Kdo residue. The overall structural architecture of the O12 CBM resembles the O9a prototype, but they share only weak sequence similarity, and the putative binding pocket for the O12 glycan is different. Removal of the CBM abrogated O-PS transport, but export was restored when the CBM was expressed in trans with the mutant CBM-deficient ABC transporter. These results demonstrate that the CBM-mediated substrate-recognition mechanism is evolutionarily conserved and can operate with glycans of widely differing structures. PMID:26934919

  4. Pleiotropic effects of the vacuolar ABC transporter MLT1 of Candida albicans on cell function and virulence.

    Khandelwal, Nitesh Kumar; Kaemmer, Philipp; Förster, Toni M; Singh, Ashutosh; Coste, Alix T; Andes, David R; Hube, Bernhard; Sanglard, Dominique; Chauhan, Neeraj; Kaur, Rupinder; d'Enfert, Christophe; Mondal, Alok Kumar; Prasad, Rajendra

    2016-06-01

    Among the several mechanisms that contribute to MDR (multidrug resistance), the overexpression of drug-efflux pumps belonging to the ABC (ATP-binding cassette) superfamily is the most frequent cause of resistance to antifungal agents. The multidrug transporter proteins Cdr1p and Cdr2p of the ABCG subfamily are major players in the development of MDR in Candida albicans Because several genes coding for ABC proteins exist in the genome of C. albicans, but only Cdr1p and Cdr2p have established roles in MDR, it is implicit that the other members of the ABC family also have alternative physiological roles. The present study focuses on an ABC transporter of C. albicans, Mlt1p, which is localized in the vacuolar membrane and specifically transports PC (phosphatidylcholine) into the vacuolar lumen. Transcriptional profiling of the mlt1∆/∆ mutant revealed a down-regulation of the genes involved in endocytosis, oxidoreductase activity, virulence and hyphal development. High-throughput MS-based lipidome analysis revealed that the Mlt1p levels affect lipid homoeostasis and thus lead to a plethora of physiological perturbations. These include a delay in endocytosis, inefficient sequestering of reactive oxygen species (ROS), defects in hyphal development and attenuated virulence. The present study is an emerging example where new and unconventional roles of an ABC transporter are being identified. PMID:27026051

  5. Molecular dynamics simulations of the bacterial ABC transporter SAV1866 in the closed form.

    St-Pierre, Jean-François; Bunker, Alex; Róg, Tomasz; Karttunen, Mikko; Mousseau, Normand

    2012-03-01

    The ATP binding cassette (ABC) transporter family of proteins contains members involved in ATP-mediated import or export of ligands at the cell membrane. For the case of exporters, the translocation mechanism involves a large-scale conformational change that involves a clothespin-like motion from an inward-facing open state, able to bind ligands and adenosine triphosphate (ATP), to an outward-facing closed state. Our work focuses on SAV1866, a bacterial member of the ABC transporter family for which the structure is known for the closed state. To evaluate the ability of this protein to undergo conformational changes at physiological temperature, we first performed conventional molecular dynamics (MD) on the cocrystallized adenosine diphosphate (ADP)-bound structure and on a nucleotide-free structure. With this assessment of SAV1866's stability, conformational changes were induced by steered molecular dynamics (SMD), in which the nucleotide binding domains (NBD) were pushed apart, simulating the ATP hydrolysis energy expenditure. We found that the transmembrane domain is not easily perturbed by large-scale motions of the NBDs. PMID:22339391

  6. ABC transporters in Mycoplasma hyopneumoniae and Mycoplasma synoviae: insights into evolution and pathogenicity

    Marisa Fabiana Nicolás

    2007-01-01

    Full Text Available ABC transporters represent one of the largest superfamilies of active membrane transport proteins (MTPs with a highly conserved ATPase domain that binds and hydrolyzes ATP, supplying energy for the uptake of a variety of nutrients and for the extrusion of drugs and metabolic wastes. The complete genomes of a non-pathogenic (J and pathogenic (7448 strain of Mycoplasma hyopneumoniae, as well as of a pathogenic (53 strain of Mycoplasma synoviae have been recently sequenced. A detailed study revealed a high percentage of CDSs encoding MTPs in M. hyopneumoniae strains J (13.4%, 7448 (13.8%, and in M. synoviae 53 (11.2%, and the ABC systems represented from 85.0 to 88.6% of those CDSs. Uptake systems are mainly involved in cell nutrition and some might be associated with virulence. Exporter systems include both drug and multidrug resistant systems (MDR, which may represent mechanisms of resistance to toxic molecules. No relation was found between the phylogeny of the ATPase domains and the lifestyle or pathogenicity of Mycoplasma, but several proteins, potentially useful as targets for the control of infections, were identified.

  7. 1.55 Å Structure of the Ectoine Binding Protein TeaA of the Osmoregulated TRAP-Transporter TeaABC from Halomonas elongata

    Kuhlmann, Sonja I.; Terwisscha van Scheltinga, Anke C; Bienert, Ralf; Kunte, Hans-Jörg; Ziegler, Christine

    2008-01-01

    TeaABC from the moderate halophilic bacterium Halomonas elongata belongs to the tripartite ATP-independent periplasmic transporters (TRAP-T), a family of secondary transporters functioning in conjunction with periplasmic substrate binding proteins. TeaABC facilitates the uptake of the compatible solutes ectoine and hydroxyectoine that are accumulated in the cytoplasm under hyperosmotic stress to protect the cell from dehydration. TeaABC is the only known TRAP-T activated by osmotic stress. Cu...

  8. Using mass spectrometry for identification of ABC transporters from Xanthomonas citri and mutants expressed in different growth conditions

    Faria, J.N.; Balan, A. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil); Paes Leme, A.F. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil)

    2012-07-01

    Full text: Xanthomonas citri is a phytopathogenic bacterium that infects citrus plants causing significant losses for the economy. In our group, we have focused on the identification and characterization of ABC transport proteins of this bacterium, in order to determinate their function for growth in vitro and in vivo, during infection. ABC transporters represent one of the largest families of proteins, which transport since small molecules as ions up to oligopeptides and sugars. In prokaryotic cells many works have reported the ABC transport function in pathogenesis, resistance, biofilm formation, infectivity and DNA repair, but until our knowledge, there is no data related to these transporters and X. citri. So, In order to determinate which transporters are expressed in X. citri, we started a proteomic analysis based on mono and bi-dimensional gels associated to mass spectrometry analyses. After growing X. citri and two different mutants deleted for ssuA and nitA genes in LB and minimum media, cellular extracts were obtained and used for preparation of mono and bi-dimensional gels. Seven bands covering the expected mass of ABC transporter components (20 kDa to 50 kDa) in SDS-PAGE were cut off the gel, treated with trypsin and submitted to the MS for protein identification. The results of 2D gels were good enough and will serve as a standard for development of similar experiments in large scale. (author)

  9. Using mass spectrometry for identification of ABC transporters from Xanthomonas citri and mutants expressed in different growth conditions

    Full text: Xanthomonas citri is a phytopathogenic bacterium that infects citrus plants causing significant losses for the economy. In our group, we have focused on the identification and characterization of ABC transport proteins of this bacterium, in order to determinate their function for growth in vitro and in vivo, during infection. ABC transporters represent one of the largest families of proteins, which transport since small molecules as ions up to oligopeptides and sugars. In prokaryotic cells many works have reported the ABC transport function in pathogenesis, resistance, biofilm formation, infectivity and DNA repair, but until our knowledge, there is no data related to these transporters and X. citri. So, In order to determinate which transporters are expressed in X. citri, we started a proteomic analysis based on mono and bi-dimensional gels associated to mass spectrometry analyses. After growing X. citri and two different mutants deleted for ssuA and nitA genes in LB and minimum media, cellular extracts were obtained and used for preparation of mono and bi-dimensional gels. Seven bands covering the expected mass of ABC transporter components (20 kDa to 50 kDa) in SDS-PAGE were cut off the gel, treated with trypsin and submitted to the MS for protein identification. The results of 2D gels were good enough and will serve as a standard for development of similar experiments in large scale. (author)

  10. ABC transporters affect the elimination and toxicity of CdTe quantum dots in liver and kidney cells.

    Chen, Mingli; Yin, Huancai; Bai, Pengli; Miao, Peng; Deng, Xudong; Xu, Yingxue; Hu, Jun; Yin, Jian

    2016-07-15

    This paper aimed to investigate the role of adenosine triphosphate-binding cassette (ABC) transporters on the efflux and the toxicity of nanoparticles in liver and kidney cells. In this study, we synthesized CdTe quantum dots (QDs) that were monodispersed and emitted green fluorescence (maximum peak at 530nm). Such QDs tended to accumulate in human hepatocellular carcinoma cells (HepG2), human kidney cells 2 (HK-2), and Madin-Darby canine kidney (MDCK) cells, and cause significant toxicity in all the three cell lines. Using specific inhibitors and inducers of P-glycoprotein (Pgp) and multidrug resistance associated proteins (Mrps), the cellular accumulation and subsequent toxicity of QDs in HepG2 and HK-2 cells were significantly affected, while only slight changes appeared in MDCK cells, corresponding well with the functional expressions of ABC transporters in cells. Moreover, treatment of QDs caused concentration- and time- dependent induction of ABC transporters in HepG2 and HK-2 cells, but such phenomenon was barely found in MDCK cells. Furthermore, the effects of CdTe QDs on ABC transporters were found to be greater than those of CdCl2 at equivalent concentrations of cadmium, indicating that the effects of QDs should be a combination of free Cd(2+) and specific properties of QDs. Overall, these results indicated a strong dependence between the functional expressions of ABC transporters and the efflux of QDs, which could be an important reason for the modulation of QDs toxicity by ABC transporters. PMID:27131644

  11. Diversity and evolution of ABC proteins in mycorrhiza-forming fungi

    Kohler, Annegret; Martin, Francis; Asiegbu, Fred O

    2015-01-01

    Background Transporter proteins are predicted to have an important role in the mycorrhizal symbiosis, due to the fact that this type of an interaction between plants and fungi requires a continuous nutrient and signalling exchange. ABC transporters are one of the large groups of transporter proteins found both in plants and in fungi. The crucial role of plant ABC transporters in the formation of the mycorrhizal symbiosis has been demonstrated recently. Some of the fungal ABC transporter-encod...

  12. Genome-wide analysis of the ATP-binding cassette (ABC) transporter gene family in the silkworm, Bombyx mori.

    Xie, Xiaodong; Cheng, Tingcai; Wang, Genhong; Duan, Jun; Niu, Weihuan; Xia, Qingyou

    2012-07-01

    The ATP-binding cassette (ABC) superfamily is a larger protein family with diverse physiological functions in all kingdoms of life. We identified 53 ABC transporters in the silkworm genome, and classified them into eight subfamilies (A-H). Comparative genome analysis revealed that the silkworm has an expanded ABCC subfamily with more members than Drosophila melanogaster, Caenorhabditis elegans, or Homo sapiens. Phylogenetic analysis showed that the ABCE and ABCF genes were highly conserved in the silkworm, indicating possible involvement in fundamental biological processes. Five multidrug resistance-related genes in the ABCB subfamily and two multidrug resistance-associated-related genes in the ABCC subfamily indicated involvement in biochemical defense. Genetic variation analysis revealed four ABC genes that might be evolving under positive selection. Moreover, the silkworm ABCC4 gene might be important for silkworm domestication. Microarray analysis showed that the silkworm ABC genes had distinct expression patterns in different tissues on day 3 of the fifth instar. These results might provide new insights for further functional studies on the ABC genes in the silkworm genome. PMID:22311044

  13. 1.55 Å Structure of the Ectoine Binding Protein TeaA of the Osmoregulated TRAP-Transporter TeaABC from Halomonas elongata

    Kuhlmann, Sonja I.; Terwisscha van Scheltinga, Anke C.; Bienert, Ralf; Kunte, Hans-Jörg; Ziegler, Christine

    2008-01-01

    TeaABC from the moderate halophilic bacterium Halomonas elongata belongs to the tripartite ATP-independent periplasmic transporters (TRAP-T), a family of secondary transporters functioning in conjunction with periplasmic substrate binding proteins. TeaABC facilitates the uptake of the compatible sol

  14. ABC- and SLC-Transporters in Murine and Bovine Mammary Epithelium - Effects of Prochloraz

    Yagdiran, Yagmur; Oskarsson, Agneta; Knight, Christopher H.; Tallkvist, Jonas

    2016-01-01

    Some chemicals are ligands to efflux transporters which may result in high concentrations in milk. Limited knowledge is available on the influence of maternal exposure to chemicals on the expression and function of transporters in the lactating mammary gland. We determined gene expression of ABC and SLC transporters in murine mammary tissue of different gestation and lactation stages, in murine mammary cells (HC11) featuring resting and secreting phenotypes and in bovine mammary tissue and cells (BME-UV). Effects on transporter expression and function of the imidazole fungicide prochloraz, previously reported to influence BCRP in mammary cells, was investigated on transporter expression and function in the two cell lines. Transporters studied were BCRP, MDR1, MRP1, OATP1A5/OATP1A2, OCTN1 and OCT1. Gene expressions of BCRP and OCT1 in murine mammary glands were increased during gestation and lactation, whereas MDR1, MRP1, OATP1A5 and OCTN1 were decreased, compared to expressions in virgins. All transporters measured in mammary glands of mice were detected in bovine mammary tissue and in HC11 cells, while only MDR1 and MRP1 were detected in BME-UV cells. Prochloraz treatment induced MDR1 gene and protein expression in both differentiated HC11 and BME-UV cells and increased protein function in HC11 cells, resulting in decreased accumulation of the MDR1 substrate digoxin. In conclusion, our results demonstrate that murine (HC11) and bovine (BME-UV) mammary epithelial cells can be applied to characterize expression and function of transporters as well as effects of contaminants on the mammary transporters. An altered expression, induced by a drug or toxic chemical, on any of the transporters expressed in the mammary epithelial cells during lactation may modulate the well-balanced composition of nutrients and/or secretion of contaminants in milk with potential adverse effects on breast-fed infants and dairy consumers. PMID:27028005

  15. A TatABC-type Tat translocase is required for unimpaired aerobic growth of Corynebacterium glutamicum ATCC13032.

    Dan Oertel

    Full Text Available The twin-arginine translocation (Tat system transports folded proteins across the cytoplasmic membrane of bacteria and the thylakoid membrane of plant chloroplasts. Escherichia coli and other Gram-negative bacteria possess a TatABC-type Tat translocase in which each of the three inner membrane proteins TatA, TatB, and TatC performs a mechanistically distinct function. In contrast, low-GC Gram-positive bacteria, such as Bacillus subtilis, use a TatAC-type minimal Tat translocase in which the TatB function is carried out by a bifunctional TatA. In high-GC Gram-positive Actinobacteria, such as Mycobacterium tuberculosis and Corynebacterium glutamicum, tatA, tatB, and tatC genes can be identified, suggesting that these organisms, just like E. coli, might use TatABC-type Tat translocases as well. However, since contrary to this view a previous study has suggested that C. glutamicum might in fact use a TatAC translocase with TatB only playing a minor role, we reexamined the requirement of TatB for Tat-dependent protein translocation in this microorganism. Under aerobic conditions, the misassembly of the Rieske iron-sulfur protein QcrA was identified as a major reason for the severe growth defect of Tat-defective C. glutamicum mutant strains. Furthermore, our results clearly show that TatB, besides TatA and TatC, is strictly required for unimpaired aerobic growth. In addition, TatB was also found to be essential for the secretion of a heterologous Tat-dependent model protein into the C. glutamicum culture supernatant. Together with our finding that expression of the C. glutamicum TatB in an E. coli ΔtatB mutant strain resulted in the formation of an active Tat translocase, our results clearly indicate that a TatABC translocase is used as the physiologically relevant functional unit for Tat-dependent protein translocation in C. glutamicum and, most likely, also in other TatB-containing Actinobacteria.

  16. A Bacitracin-Resistant Bacillus subtilis Gene Encodes a Homologue of the Membrane-Spanning Subunit of the Bacillus licheniformis ABC Transporter

    Ohki, Reiko; Tateno, Kozue; Okada, Youji; Okajima, Haruo; Asai, Kei; Sadaie, Yoshito; Murata, Makiko; Aiso, Toshiko

    2003-01-01

    Bacitracin is a peptide antibiotic nonribosomally produced by Bacillus licheniformis. The bcrABC genes which confer bacitracin resistance to the bacitracin producer encode ATP binding cassette (ABC) transporter proteins, which are hypothesized to pump out bacitracin from the cells. Bacillus subtilis 168, which has no bacitracin synthesizing operon, has several genes homologous to bcrABC. It was found that the disruption of ywoA, a gene homologous to bcrC, resulted in hypersensitivity to bacit...

  17. Imaging of ABC transporters with positron emission tomography – new insights from the development of new radiotracers

    Adenosine triphosphate-binding cassette (ABC) transporters, such as P-glycoprotein (Pgp), breast cancer resistance protein (BCRP) and multidrug resistance-associated protein 1 (MRP1), form through their ability to transport a multitude of chemically unrelated compounds across cellular membranes a powerful physiological defense system which protects the body from exposure to xenobiotics. Historically, ABC transporters have been implicated in multidrug resistance (MDR) in cancer, but more recent evidence suggests that changes in ABC transporter function and expression at the blood-brain barrier (BBB) occur in several neurological disorders, such as epilepsy, Alzheimer's and Parkinson's disease. To better understand the role of ABC transporters in disease a tool is needed to assess their function and expression in-vivo. The nuclear imaging technique positron emission tomography (PET), which enables measurement of tissue concentration levels of radiolabeled compounds, has great potential for studying ABC transporters in-vivo, provided the availability of suitable radiotracers and imaging protocols. In this thesis the interaction of new radiotracers with Pgp and BCRP was investigated in order to assess their suitability for PET imaging. The radiolabeled Pgp inhibitors [11C]tariquidar and [11C]elacridar had been initially developed to visualize Pgp density at the BBB but had failed to provide useful, quantifiable brain PET signals. In this thesis, the suitability of [11C]tariquidar and [11C]elacridar to visualize Pgp density in tumors was investigated. An experimental murine tumor model comprising a wild-type and a Pgp overexpressing variant was developed and binding characteristics of [11C]tariquidar and [11C]elacridar as well as two other newly developed PET tracers ([11C]MC113 and [11C]HM30181) were systematically evaluated. These studies showed, that contrary to the BBB, radiolabeled Pgp inhibitors were able to assess Pgp density in tumors, which may find

  18. Efficient and stable reconstitution of the ABC transporter BmrA for solid-state NMR studies.

    Kunert, Britta; Gardiennet, Carole; Lacabanne, Denis; Calles-Garcia, Daniel; Falson, Pierre; Jault, Jean-Michel; Meier, Beat H; Penin, François; Böckmann, Anja

    2014-01-01

    We present solid-state NMR sample preparation and first 2D spectra of the Bacillus subtilis ATP-binding cassette (ABC) transporter BmrA, a membrane protein involved in multidrug resistance. The homodimeric 130-kDa protein is a challenge for structural characterization due to its membrane-bound nature, size, inherent flexibility and insolubility. We show that reconstitution of this protein in lipids from Bacillus subtilis at a lipid-protein ratio of 0.5 w/w allows for optimal protein insertion in lipid membranes with respect to two central NMR requirements, high signal-to-noise in the spectra and sample stability over a time period of months. The obtained spectra point to a well-folded protein and a highly homogenous preparation, as witnessed by the narrow resonance lines and the signal dispersion typical for the expected secondary structure distribution of BmrA. This opens the way for studies of the different conformational states of the transporter in the export cycle, as well as on interactions with substrates, via chemical-shift fingerprints and sequential resonance assignments. PMID:25988146

  19. ABC transporter functions as a pacemaker for sequestration of plant glucosides in leaf beetles.

    Strauss, Anja S; Peters, Sven; Boland, Wilhelm; Burse, Antje

    2013-01-01

    Plant-herbivore interactions dominate the planet's terrestrial ecology. When it comes to host-plant specialization, insects are among the most versatile evolutionary innovators, able to disarm multiple chemical plant defenses. Sequestration is a widespread strategy to detoxify noxious metabolites, frequently for the insect's own benefit against predation. In this study, we describe the broad-spectrum ATP-binding cassette transporter CpMRP of the poplar leaf beetle, Chrysomela populi as the first candidate involved in the sequestration of phytochemicals in insects. CpMRP acts in the defensive glands of the larvae as a pacemaker for the irreversible shuttling of pre-selected metabolites from the hemolymph into defensive secretions. Silencing CpMRP in vivo creates a defenseless phenotype, indicating its role in the secretion process is crucial. In the defensive glands of related leaf beetle species, we identified sequences similar to CpMRP and assume therefore that exocrine gland-based defensive strategies, evolved by these insects to repel their enemies, rely on ABC transporters as a key element. DOI: http://dx.doi.org/10.7554/eLife.01096.001. PMID:24302568

  20. Phenotypic, Proteomic, and Genomic Characterization of a Putative ABC-Transporter Permease Involved in Listeria monocytogenes Biofilm Formation

    Zhu, Xinna; Liu, Weibing; Lametsch, René;

    2011-01-01

    The foodborne pathogen Listeria monocytogenes is able to form biofilms in food processing environments. Previously, we have reported that an lm.G_1771 gene (encoding a putative ABC-transporter permease) was involved in negative regulation of L. monocytogenes biofilm formation using LM-49, a biofilm......-enhanced mutant isolated on Tn917 mutagenesis (AEM 2008 p.7675–7683). Here, the possible action of this ABC-transporter permease in L. monocytogenes biofilm formation was characterized by phenotypic, proteomic, and genomic analyses using an lm.G_1771 gene deletant (Δ1771). The Δ1771 mutant exhibited the same...... enhanced ability for biofilm formation as the LM-49 strain using a crystal violet staining assay. DNA microarrays and two-dimensional gel electrophoresis revealed 49 and 11 differentially expressed (twofold or more) genes or proteins in Δ1771, respectively. The transcriptomics study indicated that lm...

  1. Pharmacogenetics of human ABC transporter ABCC11:new insights into apocrine gland growth and metabolite secretion

    Toshihisa eIshikawa

    2013-01-01

    Full Text Available Cell secretion is an important physiological process that ensures smooth metabolic activities and tissue repair as well as growth and immunological functions in the body. Apocrine secretion occurs when the secretory process is accomplished with a partial loss of cell cytoplasm. The secretory materials are contained within secretory vesicles and are released during secretion as cytoplasmic fragments into the glandular lumen or interstitial space. The recent finding that the nonsynonymous SNP 538G>A (rs17822931; Gly180Arg in the ABCC11 gene determines the type of earwax in humans has shed light on the novel function of this ABC transporter in apocrine glands. The wild type (Gly180 of ABCC11 is associated with wet-type earwax, axillary osmidrosis, and colostrum secretion from the mammary gland as well as the potential risk of mastopathy. Furthermore, the SNP (538G>A in the ABCC11 gene is suggested to be a clinical biomarker for the prediction of chemotherapeutic efficacy. The aim of this review article is to provide an overview on the discovery and characterization of genetic polymorphisms in the human ABCC11 gene and to explain the impact of ABCC11 538G>A on the apocrine phenotype as well as the anthropological aspect of this SNP in the ABCC11 gene and patients’ response to nucleoside-based chemotherapy.

  2. Lysophosphatidylinositol: a novel link between ABC transporters and G-protein-coupled receptors.

    Ruban, Emily L; Ferro, Riccardo; Arifin, Syamsul Ahmad; Falasca, Marco

    2014-10-01

    Lysophosphatidylinositol (LPI) is a well-known bioactive lipid that is able to activate signalling cascades relevant to cell proliferation, migration, survival and tumorigenesis. Our previous work suggested that LPI is involved in cancer progression since it can be released in the medium of Ras-transformed fibroblasts and can function as an autocrine modulator of cell growth. Different research groups have established that LPI is the specific and functional ligand for G-protein-coupled receptor 55 (GPR55) and that this GPR55-LPI axis is able to activate signalling cascades that are relevant for different cell functions. Work in our laboratory has recently unravelled an autocrine loop, by which LPI synthesized by cytosolic phospholipase A₂ (cPLA₂) is pumped out of the cell by ATP-binding cassette (ABC) transporter C1 (ABCC1)/multidrug resistance protein 1 (MRP1), initiating a signalling cascade downstream of GPR55. Our current work suggests that blockade of this pathway may represent a novel strategy to inhibit cancer cell proliferation. PMID:25233417

  3. Identification and characterization of an iron ABC transporter operon in Gluconacetobacter diazotrophicus Pal 5.

    Urzúa, Lucia Soto; Vázquez-Candanedo, Ada P; Sánchez-Espíndola, Adriana; Ramírez, Carlos Ávila; Baca, Beatriz E

    2013-06-01

    Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium and endophyte of sugarcane. We have cloned and sequenced the genes coding for the components of the iron ABC-type acquisition system of G. diazotrophicus. Sequence analysis revealed three ORFs, (feuA, feuB, and feuC) organized as an operon and encoding polypeptides of 346 (38 kDa), 342 (34.2 kDa), and 240 (26 kDa) amino acids, respectively. The deduced translation products of the feu operon showed similarity with a periplasmic solute-binding protein (FeuA), permease (FeuB), and ATPase (FeuC) involved in Fe transport. The role of FeuB in the survival of G. diazotrophicus under iron depletion was evaluated by comparing the ability of wild-type and FeuB-Km(R) -mutant strains in a medium without iron supplementation and in a medium containing 2, 2'-dipyridyl (DP). Growth of the mutant was affected in the medium containing DP. The operon was expressed at higher levels in cells depleted for iron than in those that contained the metal. A decrease in nitrogenase activity was observed with the FeuB-Km(R) -mutant strain that with the wild-type under iron deficiency conditions, suggesting that the Feu operon play role in Fe nutrition of G. diazotrophicus. PMID:23624722

  4. Crystal structure of the phosphate-binding protein (PBP-1) of an ABC-type phosphate transporter from Clostridium perfringens

    Gonzalez, Daniel; Richez, Magali; Bergonzi, Celine; Chabriere, Eric; Elias, Mikael

    2014-01-01

    Phosphate limitation is an important environmental stress that affects the metabolism of various organisms and, in particular, can trigger the virulence of numerous bacterial pathogens. Clostridium perfringens, a human pathogen, is one of the most common causes of enteritis necroticans, gas gangrene and food poisoning. Here, we focused on the high affinity phosphate-binding protein (PBP-1) of an ABC-type transporter, responsible for cellular phosphate uptake. We report the crystal structure (...

  5. A two-component system regulates the expression of an ABC transporter for xylo-oligosaccharides in Geobacillus stearothermophilus.

    Shulami, Smadar; Zaide, Galia; Zolotnitsky, Gennady; Langut, Yael; Feld, Geoff; Sonenshein, Abraham L; Shoham, Yuval

    2007-02-01

    Geobacillus stearothermophilus T-6 utilizes an extensive and highly regulated hemicellulolytic system. The genes comprising the xylanolytic system are clustered in a 39.7-kb chromosomal segment. This segment contains a 6-kb transcriptional unit (xynDCEFG) coding for a potential two-component system (xynDC) and an ATP-binding cassette (ABC) transport system (xynEFG). The xynD promoter region contains a 16-bp inverted repeat resembling the operator site for the xylose repressor, XylR. XylR was found to bind specifically to this sequence, and binding was efficiently prevented in vitro in the presence of xylose. The ABC transport system was shown to comprise an operon of three genes (xynEFG) that is transcribed from its own promoter. The nonphosphorylated fused response regulator, His6-XynC, bound to a 220-bp fragment corresponding to the xynE operator. DNase I footprinting analysis showed four protected zones that cover the -53 and the +34 regions and revealed direct repeat sequences of a GAAA-like motif. In vitro transcriptional assays and quantitative reverse transcription-PCR demonstrated that xynE transcription is activated 140-fold in the presence of 1.5 microM XynC. The His6-tagged sugar-binding lipoprotein (XynE) of the ABC transporter interacted with different xylosaccharides, as demonstrated by isothermal titration calorimetry. The change in the heat capacity of binding (DeltaCp) for XynE with xylotriose suggests a stacking interaction in the binding site that can be provided by a single Trp residue and a sugar moiety. Taken together, our data show that XynEFG constitutes an ABC transport system for xylo-oligosaccharides and that its transcription is negatively regulated by XylR and activated by the response regulator XynC, which is part of a two-component sensing system. PMID:17142383

  6. Mutational Analysis of Intracellular Loops Identify Cross Talk with Nucleotide Binding Domains of Yeast ABC Transporter Cdr1p

    Shah, Abdul Haseeb; Rawal, Manpreet Kaur; Dhamgaye, Sanjiveeni; Komath, Sneha Sudha; Saxena, Ajay Kumar; Prasad, Rajendra

    2015-01-01

    The ABC transporter Cdr1 protein (Cdr1p) of Candida albicans, which plays a major role in antifungal resistance, has two transmembrane domains (TMDs) and two nucleotide binding domains (NBDs) that are interconnected by extracellular (ECLs) and intracellular (ICLs) loops. To examine the communication interface between the NBDs and ICLs of Cdr1p, we subjected all four ICLs to alanine scanning mutagenesis, replacing each of the 85 residues with an alanine. The resulting ICL mutant library was an...

  7. The ABC transporter BcatrB from Botrytis cinerea exports camalexin and is a virulence factor on Arabidopsis thaliana

    Stefanato, Francesca L.; Abou-Mansour, Eliane; Buchala, Antony; Kretschmer, Matthias; Mosbach, Andreas; Hahn, Matthias; Bochet, Christian G.; Métraux, Jean-Pierre; Schoonbeek, Henk-jan

    2009-01-01

    Arabidopsis thaliana is known to produce the phytoalexin camalexin in response to abiotic and biotic stress. Here we studied the mechanisms of tolerance to camalexin in the fungus Botrytis cinerea, a necrotrophic pathogen of A. thaliana. Exposure of B. cinerea to camalexin induces expression of BcatrB, an ABC transporter that functions in the efflux of fungitoxic compounds. B. cinerea inoculated on wild-type A. thaliana plants yields smaller lesions than on camalexin-deficient A. thaliana mut...

  8. Identification of the tliDEF ABC Transporter Specific for Lipase in Pseudomonas fluorescens SIK W1

    Ahn, Jung Hoon; Pan, Jae Gu; Rhee, Joon Shick

    1999-01-01

    Pseudomonas fluorescens, a gram-negative psychrotrophic bacterium, secretes a thermostable lipase into the extracellular medium. In our previous study, the lipase of P. fluorescens SIK W1 was cloned and expressed in Escherichia coli, but it accumulated as inactive inclusion bodies. Amino acid sequence analysis of the lipase revealed a potential C-terminal targeting sequence recognized by the ATP-binding cassette (ABC) transporter. The genetic loci around the lipase gene were searched, and a s...

  9. Role of ABC and Solute Carrier Transporters in the Placental Transport of Lamivudine.

    Ceckova, Martina; Reznicek, Josef; Ptackova, Zuzana; Cerveny, Lukas; Müller, Fabian; Kacerovsky, Marian; Fromm, Martin F; Glazier, Jocelyn D; Staud, Frantisek

    2016-09-01

    Lamivudine is one of the antiretroviral drugs of choice for the prevention of mother-to-child transmission (MTCT) in HIV-positive women. In this study, we investigated the relevance of drug efflux transporters P-glycoprotein (P-gp) (MDR1 [ABCB1]), BCRP (ABCG2), MRP2 (ABCC2), and MATE1 (SLC47A1) for the transmembrane transport and transplacental transfer of lamivudine. We employed in vitro accumulation and transport experiments on MDCK cells overexpressing drug efflux transporters, in situ-perfused rat term placenta, and vesicular uptake in microvillous plasma membrane (MVM) vesicles isolated from human term placenta. MATE1 significantly accelerated lamivudine transport in MATE1-expressing MDCK cells, whereas no transporter-driven efflux of lamivudine was observed in MDCK-MDR1, MDCK-MRP2, and MDCK-BCRP monolayers. MATE1-mediated efflux of lamivudine appeared to be a low-affinity process (apparent Km of 4.21 mM and Vmax of 5.18 nmol/mg protein/min in MDCK-MATE1 cells). Consistent with in vitro transport studies, the transplacental clearance of lamivudine was not affected by P-gp, BCRP, or MRP2. However, lamivudine transfer across dually perfused rat placenta and the uptake of lamivudine into human placental MVM vesicles revealed pH dependency, indicating possible involvement of MATE1 in the fetal-to-maternal efflux of the drug. To conclude, placental transport of lamivudine does not seem to be affected by P-gp, MRP2, or BCRP, but a pH-dependent mechanism mediates transport of lamivudine in the fetal-to-maternal direction. We suggest that MATE1 might be, at least partly, responsible for this transport. PMID:27401571

  10. Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation

    Teixeira Miguel C; Godinho Cláudia P; Cabrito Tânia R; Mira Nuno P; Sá-Correia Isabel

    2012-01-01

    Abstract Background The understanding of the molecular basis of yeast tolerance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. A set of 21 genes encoding multidrug transporters from the ATP-Binding Cassette (ABC) Superfamily and Major Facilitator Superfamily (MFS) in S. cerevisiae were scrutinized for a role in ethanol stress resistance. Results A yeast multidrug resistance ABC transporter encoded by the PDR18 gene...

  11. Active transmembrane drug transport in microgravity: a validation study using an ABC transporter model [v1; ref status: indexed, http://f1000r.es/41n

    Sergi Vaquer

    2014-08-01

    Full Text Available Abstract Microgravity has been shown to influence the expression of ABC (ATP-Binding Cassette transporters in bacteria, fungi and mammals, but also to modify the activity of certain cellular components with structural and functional similarities to ABC transporters. Changes in activity of ABC transporters could lead to important metabolic disorders and undesired pharmacological effects during spaceflights. However, no current means exist to study the functionality of these transporters in microgravity. To this end, a Vesicular Transport Assay® (Solvo Biotechnology, Hungary was adapted to evaluate multi-drug resistance-associated protein 2 (MRP2 trans-membrane estradiol-17-β-glucuronide (E17βG transport activity, when activated by adenosine-tri-phosphate (ATP during parabolic flights. Simple diffusion, ATP-independent transport and benzbromarone inhibition were also evaluated. A high accuracy engineering system was designed to perform, monitor and synchronize all procedures. Samples were analysed using a validated high sensitivity drug detection protocol. Experiments were performed in microgravity during parabolic flights, and compared to 1g on ground results using identical equipment and procedures in all cases. Our results revealed that sufficient equipment accuracy and analytical sensitivity were reached to detect transport activity in both gravitational conditions. Additionally, transport activity levels of on ground samples were within commercial transport standards, proving the validity of the methods and equipment used. MRP2 net transport activity was significantly reduced in microgravity, so was signal detected in simple diffusion samples. Ultra-structural changes induced by gravitational stress upon vesicle membranes or transporters could explain the current results, although alternative explanations are possible. Further research is needed to provide a conclusive answer in this regard. Nevertheless, the present validated technology

  12. The Schizosaccharomyces pombe mam1 gene encodes an ABC transporter mediating secretion of M-factor

    Christensen, P U; Davey, William John; Nielsen, O;

    1997-01-01

    In the fission yeast Schizosaccharomyces pombe, cells of opposite mating type communicate via diffusible peptide pheromones prior to mating. We have cloned the S. pombe mam1 gene, which encodes a 1336-amino acid protein belonging to the ATP-binding cassette (ABC) superfamily. The mam1 gene is onl...

  13. Tivantinib (ARQ 197) exhibits antitumor activity by directly interacting with tubulin and overcomes ABC transporter-mediated drug resistance.

    Aoyama, Aki; Katayama, Ryohei; Oh-Hara, Tomoko; Sato, Shigeo; Okuno, Yasushi; Fujita, Naoya

    2014-12-01

    Tivantinib (ARQ197) was first reported as a highly selective inhibitor of c-MET and is currently being investigated in a phase III clinical trial. However, as recently reported by us and another group, tivantinib showed cytotoxic activity independent of cellular c-MET status and also disrupted microtubule dynamics. To investigate if tivantinib exerts its cytotoxic activity by disrupting microtubules, we quantified polymerized tubulin in cells and xenograft tumors after tivantinib treatment. Consistent with our previous report, tivantinib reduced tubulin polymerization in cells and in mouse xenograft tumors in vivo. To determine if tivantinib directly binds to tubulin, we performed an in vitro competition assay. Tivantinib competitively inhibited colchicine but not vincristine or vinblastine binding to purified tubulin. These results imply that tivantinib directly binds to the colchicine binding site of tubulin. To predict the binding mode of tivantinib with tubulin, we performed computer simulation of the docking pose of tivantinib with tubulin using GOLD docking program. Computer simulation predicts tivantinib fitted into the colchicine binding pocket of tubulin without steric hindrance. Furthermore, tivantinib showed similar IC50 values against parental and multidrug-resistant cells. In contrast, other microtubule-targeting drugs, such as vincristine, paclitaxel, and colchicine, could not suppress the growth of cells overexpressing ABC transporters. Moreover, the expression level of ABC transporters did not correlate with the apoptosis-inducing ability of tivantinib different from other microtubule inhibitor. These results suggest that tivantinib can overcome ABC transporter-mediated multidrug-resistant tumor cells and is potentially useful against various tumors. PMID:25313010

  14. The ABC transporter ABCG29 is involved in H2O2 tolerance and biocontrol traits in the fungus Clonostachys rosea.

    Dubey, Mukesh; Jensen, Dan Funck; Karlsson, Magnus

    2016-04-01

    For successful biocontrol interactions, biological control organisms must tolerate toxic metabolites produced by themselves or plant pathogens during mycoparasitic/antagonistic interactions, by host plant during colonization of the plant, and xenobiotics present in the environment. ATP-binding cassette (ABC) transporters can play a significant role in tolerance of toxic compounds by mediating active transport across the cellular membrane. This paper reports on functional characterization of an ABC transporter ABCG29 in the biocontrol fungus Clonostachys rosea strain IK726. Gene expression analysis showed induced expression of abcG29 during exposure to the Fusarium spp. mycotoxin zearalenone (ZEA) and the fungicides Cantus, Chipco Green and Apron. Expression of abcG29 in C. rosea was significantly higher during C. rosea-C. rosea (Cr-Cr) interaction or in exposure to C. rosea culture filtrate for 2 h, compared to interaction with Fusarium graminearum or 2 h exposure to F. graminearum culture filtrate. In contrast with gene expression data, ΔabcG29 strains did not display reduced tolerance towards ZEA, fungicides or chemical agents known for inducing oxidative, cell wall or osmotic stress, compared to C. rosea WT. The exception was a significant reduction in tolerance to H2O2 (10 mM) in ΔabcG29 strains when conidia were used as an inoculum. The antagonistic ability of ΔabcG29 strains towards F. graminearum, Fusarium oxysporum or Botrytis cinerea in dual plate assays were not different compared with WT. However, in biocontrol assays ΔabcG29 strains displayed reduced ability to protect Arabidopsis thaliana leaves from B. cinerea, and barley seedling from F. graminearum as measured by an A. thaliana detached leaf assay and a barley foot rot disease assay, respectively. These data show that the ABCG29 is dispensable for ZEA and fungicides tolerance, and antagonism but not H2O2 tolerance and biocontrol effects in C. rosea. PMID:26520102

  15. Heme Transfer from Streptococcal Cell Surface Protein Shp to HtsA of Transporter HtsABC

    Liu, Mengyao; Lei, Benfang

    2005-01-01

    Human pathogen group A streptococcus (GAS) can take up heme from host heme-containing proteins as a source of iron. Little is known about the heme acquisition mechanism in GAS. We recently identified a streptococcal cell surface protein (designated Shp) and the lipoprotein component (designated HtsA) of an ATP-binding cassette (ABC) transporter made by GAS as heme-binding proteins. In an effort to delineate the molecular mechanism involved in heme acquisition by GAS, heme-free Shp (apo-Shp) a...

  16. Efficient and stable reconstitution of the ABC transporter BmrA for solid-state NMR studies

    Kunert, Britta; Gardiennet, Carole; Lacabanne, Denis; Calles-Garcia, Daniel; Falson, Pierre; Jault, Jean-Michel; Meier, Beat H; Penin, François; Böckmann, Anja

    2014-01-01

    We present solid-state NMR sample preparation and first 2D spectra of the Bacillus subtilis ATP-binding cassette (ABC) transporter BmrA, a membrane protein involved in multidrug resistance. The homodimeric 130-kDa protein is a challenge for structural characterization due to its membrane-bound nature, size, inherent flexibility and insolubility. We show that reconstitution of this protein in lipids from Bacillus subtilis at a lipid-protein ratio of 0.5 w/w allows for optimal protein insertion...

  17. The Caulobacter crescentus Paracrystalline S-Layer Protein Is Secreted by an ABC Transporter (Type I) Secretion Apparatus

    Awram, Peter; Smit, John

    1998-01-01

    Caulobacter crescentus is a gram-negative bacterium that produces a two-dimensional crystalline array on its surface composed of a single 98-kDa protein, RsaA. Secretion of RsaA to the cell surface relies on an uncleaved C-terminal secretion signal. In this report, we identify two genes encoding components of the RsaA secretion apparatus. These components are part of a type I secretion system involving an ABC transporter protein. These genes, lying immediately 3′ of rsaA, were found by screen...

  18. In vitro functional characterization of BtuCD-F, the Escherichia coli ABC transporter for vitamin B-12 uptake

    Borths, EL; Poolman, B; Hvorup, RN; Locher, KP; Rees, DC; Hvorup, Rikki N.; Locher, Kaspar P.; Rees, Douglas C.

    2005-01-01

    BtuCD is an ATP binding cassette (ABC) transporter that facilitates uptake of vitamin B-12 into the cytoplasm of Escherichia coli. The crystal structures of BtuCD and its cognate periplasmic binding protein BtuF have been recently determined. We have now explored BtuCD-F function in vitro, both in proteoliposomes and in various detergents. BtuCD reconstituted into proteoliposomes has a significant basal ATP hydrolysis rate that is stimulated by addition of BtuF and inhibited by sodium ortho-v...

  19. The abcEDCBA-Encoded ABC Transporter and the virB Operon-Encoded Type IV Secretion System of Brucella ovis Are Critical for Intracellular Trafficking and Survival in Ovine Monocyte-Derived Macrophages.

    Auricelio A Macedo

    Full Text Available Brucella ovis infection is associated with epididymitis, orchitis and infertility in rams. Most of the information available on B. ovis and host cell interaction has been generated using murine macrophages or epithelial cell lines, but the interaction between B. ovis and primary ovine macrophages has not been studied. The aim of this study was to evaluate the role of the B. ovis abcEDCBA-encoded ABC transporter and the virB operon-encoded Type IV Secretion System (T4SS during intracellular survival of B. ovis in ovine peripheral blood monocyte-derived macrophages. ΔabcBA and ΔvirB2 mutant strains were unable to survive in the intracellular environment when compared to the WT B. ovis at 48 hours post infection (hpi. In addition, these mutant strains cannot exclude the lysosomal marker LAMP1 from its vacuolar membrane, and their vacuoles do not acquire the endoplasmic reticulum marker calreticulin, which takes place in the WT B. ovis containing vacuole. Higher levels of nitric oxide production were observed in macrophages infected with WT B. ovis at 48 hpi when compared to macrophages infected with the ΔabcBA or ΔvirB2 mutant strains. Conversely, higher levels of reactive oxygen species were detected in macrophages infected with the ΔabcBA or ΔvirB2 mutant strains at 48 hpi when compared to macrophages infected with the WT strain. Our results demonstrate that B. ovis is able to persist and multiply in ovine macrophages, while ΔabcBA and ΔvirB2 mutations prevent intracellular multiplication, favor phagolysosome fusion, and impair maturation of the B. ovis vacuole towards an endoplasmic reticulum-derived compartment.

  20. Drosophila ABC Transporter DmHMT-1 Confers Tolerance to Cadmium.

    Half molecule ATP-binding cassette transporters of the HMT1(heavy metal tolerance factor 1)subfamily are required for Cd2+ tolerance in Schizosaccharomyces pombe, Caenorhabditis elegans and Chlamydomonas reinhardtii, and have homologs in other species, including plants and humans. Based on studies i...

  1. ABC Technology Development Program

    The Accelerator-Based Conversion (ABC) facility will be designed to accomplish the following mission: 'Provide a weapon's grade plutonium disposition capability in a safe, economical, and environmentally sound manner on a prudent schedule for [50] tons of weapon's grade plutonium to be disposed on in [20] years.' This mission is supported by four major objectives: provide a reliable plutonium disposition capability within the next [15] years; provide a level of safety and of safety assurance that meets or exceeds that afforded to the public by modern commercial nuclear power plants; meet or exceed all applicable federal, state, and local regulations or standards for environmental compliance; manage the program in a cost effective manner. The ABC Technology Development Program defines the technology development activities that are required to accomplish this mission. The technology development tasks are related to the following topics: blanket system; vessel systems; reactivity control systems; heat transport system components; energy conversion systems; shutdown heat transport systems components; auxiliary systems; technology demonstrations - large scale experiments

  2. Quantitative Analysis of the Relative Transcript Levels of ABC Transporter Atr Genes in Aspergillus nidulans by Real-Time Reverse Transcription-PCR Assay

    Pizeta Semighini, Camile; Marins, Mozart; Goldman, Maria Helena S.; Goldman, Gustavo Henrique

    2002-01-01

    The development of assays for quantitative analysis of the relative transcript levels of ABC transporter genes by real-time reverse transcription-PCR (RT-PCR) might provide important information about multidrug resistance in filamentous fungi. Here, we evaluate the potential of real-time RT-PCR to quantify the relative transcript levels of ABC transporter Atr genes from Aspergillus nidulans. The AtrA to AtrD genes showed different and higher levels in the presence of structurally unrelated dr...

  3. Human ABC transporter ABCG2/BCRP expression in chemoresistance: basic and clinical perspectives for molecular cancer therapeutics

    Noguchi K

    2014-02-01

    Full Text Available Kohji Noguchi, Kazuhiro Katayama, Yoshikazu Sugimoto Division of Chemotherapy, Faculty of Pharmacy, Keio University, Tokyo, Japan Abstract: Adenine triphosphate (ATP-binding cassette (ABC transporter proteins, such as ABCB1/P-glycoprotein (P-gp and ABCG2/breast cancer resistance protein (BCRP, transport various structurally unrelated compounds out of cells. ABCG2/BCRP is referred to as a “half-type” ABC transporter, functioning as a homodimer, and transports anticancer agents such as irinotecan, 7-ethyl-10-hydroxycamptothecin (SN-38, gefitinib, imatinib, methotrexate, and mitoxantrone from cells. The expression of ABCG2/BCRP can confer a multidrug-resistant phenotype on cancer cells and affect drug absorption, distribution, metabolism, and excretion in normal tissues, thus modulating the in vivo efficacy of chemotherapeutic agents. Clarification of the substrate preferences and structural relationships of ABCG2/BCRP is essential for our understanding of the molecular mechanisms underlying its effects in vivo during chemotherapy. Its single-nucleotide polymorphisms are also involved in determining the efficacy of chemotherapeutics, and those that reduce the functional activity of ABCG2/BCRP might be associated with unexpected adverse effects from normal doses of anticancer drugs that are ABCG2/BCRP substrates. Importantly, many recently developed molecular-targeted cancer drugs, such as the tyrosine kinase inhisbitors, imatinib mesylate, gefitinib, and others, can also interact with ABCG2/BCRP. Both functional single-nucleotide polymorphisms and inhibitory agents of ABCG2/BCRP modulate the in vivo pharmacokinetics and pharmacodynamics of these molecular cancer treatments, so the pharmacogenetics of ABCG2/BCRP is an important consideration in the application of molecular-targeted chemotherapies. Keywords: kinase inhibitor, SNP, single-nucleotide polymorphisms, molecular target

  4. Environmental Conditions Influence Induction of Key ABC-Transporter Genes Affecting Glyphosate Resistance Mechanism in Conyza canadensis

    Tani, Eleni; Chachalis, Demosthenis; Travlos, Ilias S.; Bilalis, Dimitrios

    2016-01-01

    Conyza canadensis has been reported to be the most frequent weed species that evolved resistance to glyphosate in various parts of the world. The objective of the present study was to investigate the effect of environmental conditions (temperature and light) on the expression levels of the EPSPS gene and two major ABC-transporter genes (M10 and M11) on glyphosate susceptible (GS) and glyphosate resistant (GR) horseweed populations, collected from several regions across Greece. Real-time PCR was conducted to determine the expression level of the aforementioned genes when glyphosate was applied at normal (1×; 533 g·a.e.·ha−1) and high rates (4×, 8×), measured at an early one day after treatment (DAT) and a later stage (four DAT) of expression. Plants were exposed to light or dark conditions, at three temperature regimes (8, 25, 35 °C). GR plants were made sensitive when exposed to 8 °C with light; those sensitized plants behaved biochemically (shikimate accumulation) and molecularly (expression of EPSPS and ABC-genes) like the GS plants. Results from the current study show the direct link between the environmental conditions and the induction level of the above key genes that likely affect the efficiency of the proposed mechanism of glyphosate resistance. PMID:27104532

  5. Environmental Conditions Influence Induction of Key ABC-Transporter Genes Affecting Glyphosate Resistance Mechanism in Conyza canadensis.

    Tani, Eleni; Chachalis, Demosthenis; Travlos, Ilias S; Bilalis, Dimitrios

    2016-01-01

    Conyza canadensis has been reported to be the most frequent weed species that evolved resistance to glyphosate in various parts of the world. The objective of the present study was to investigate the effect of environmental conditions (temperature and light) on the expression levels of the EPSPS gene and two major ABC-transporter genes (M10 and M11) on glyphosate susceptible (GS) and glyphosate resistant (GR) horseweed populations, collected from several regions across Greece. Real-time PCR was conducted to determine the expression level of the aforementioned genes when glyphosate was applied at normal (1×; 533 g·a.e.·ha(-1)) and high rates (4×, 8×), measured at an early one day after treatment (DAT) and a later stage (four DAT) of expression. Plants were exposed to light or dark conditions, at three temperature regimes (8, 25, 35 °C). GR plants were made sensitive when exposed to 8 °C with light; those sensitized plants behaved biochemically (shikimate accumulation) and molecularly (expression of EPSPS and ABC-genes) like the GS plants. Results from the current study show the direct link between the environmental conditions and the induction level of the above key genes that likely affect the efficiency of the proposed mechanism of glyphosate resistance. PMID:27104532

  6. Structural basis for the hydrolysis of ATP by a nucleotide binding subunit of an amino acid ABC transporter from Thermus thermophilus.

    Devi, Seenivasan Karthiga; Chichili, Vishnu Priyanka Reddy; Jeyakanthan, J; Velmurugan, D; Sivaraman, J

    2015-06-01

    ATP-binding cassette (ABC) transporters are a major family of small molecule transporter proteins, and their deregulation is associated with several diseases, including cancer. Here, we report the crystal structure of the nucleotide binding domain (NBD) of an amino acid ABC transporter from Thermus thermophilus (TTHA1159) in its apo form and as a complex with ADP along with functional studies. TTHA1159 is a putative arginine ABC transporter. The apo-TTHA1159 was crystallized in dimeric form, a hitherto unreported form of an apo NBD. Structural comparison of the apo and ADP-Mg(2+) complexes revealed that Phe14 of TTHA1159 undergoes a significant conformational change to accommodate ADP, and that the bound ADP interacts with the P-loop (Gly40-Thr45). Modeling of ATP-Mg(2+):TTHA1159 complex revealed that Gln86 and Glu164 are involved in water-mediated hydrogen bonding contacts and Asp163 in Mg(2+) ion-mediated hydrogen bonding contacts with the γ-phosphate of ATP, consistent with the findings of other ABC transporters. Mutational studies confirmed the necessity of each of these residues, and a comparison of the apo/ADP Mg(2+):TTHA1159 with its ATP-complex model suggests the likelihood of a key conformational change to the Gln86 side chain for ATP hydrolysis. PMID:25916755

  7. Secondary metabolites inhibiting ABC transporters and reversing resistance of cancer cells and fungi to cytotoxic and antimicrobial agents

    Michael eWink

    2012-04-01

    Full Text Available Fungal, bacterial and cancer cells can develop resistance against antifungal, antibacterial or anticancer agents. Mechanisms of resistance are complex and often multifactorial. Mechanisms include: 1. Activation of ABC transporters, such as P-gp, which pump out lipophilic compounds that have entered a cell, 2. Activation of cytochrome p450 oxidases which can oxidise lipophilic agents to make them more hydrophilic and accessible for conjugation reaction with glucuronic acid, sulphate or amino acids, and 3. Activation of glutathione transferase, which can conjugate xenobiotics. This review summarises the evidence that secondary metabolites of plants, such as alkaloids, phenolics and terpenoids can interfere with ABC transporters in cancer cells, parasites, bacteria and fungi. Among the active natural products several lipophilic terpenoids ( monoterpenes, diterpenes, triterpenes (including saponins, steroids (including cardiac glycosides and tetraterpenes but also some alkaloids (isoquinoline, protoberberine, quinoline, indole, monoterpene indole, and steroidal alkaloids function probably as competitive inhibitors of P-gp, MRP1 and BCRP in cancer cells, or efflux pumps in bacteria (NorA and fungi. More polar phenolics (phenolic acids, flavonoids, catechins, chalcones, xanthones, stilbenes, anthocyanins, tannins, anthraquinones, and naphthoquinones directly inhibit proteins forming several hydrogen and ionic bonds and thus disturbing the 3D structure of the transporters. The natural products may be interesting in medicine or agriculture as they can enhance the activity of active chemotherapeutics or pesticides or even reverse MDR, at least partially, of adapted and resistant cells. If these secondary metabolites are applied in combination with a cytotoxic or antimicrobial agent, they may reverse resistance in a synergistic fashion.

  8. Reversing multidrug resistance in breast cancer cells by silencing ABC transporter genes with nanoparticle-facilitated delivery of target siRNAs

    Li YT

    2012-06-01

    Full Text Available Yong Tsuey Li,1 Ming Jang Chua,1 Anil Philip Kunnath,1 Ezharul Hoque Chowdhury,1,21Faculty of Medicine and Health Science, International Medical University (IMU, No 126, Jalan 19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia; 2Jeffrey Cheah School of Medicine and Health Sciences, Faculty of Medicine, Nursing and Health Sciences, Monash University Kuala Lumpur, MalaysiaBackground: Multidrug resistance, a major impediment to successful cancer chemotherapy, is the result of overexpression of ATP-binding cassette (ABC transporters extruding internalized drugs. Silencing of ABC transporter gene expression with small interfering RNA (siRNA could be an attractive approach to overcome multidrug resistance of cancer, although delivery of siRNA remains a major hurdle to fully exploit the potential of siRNA-based therapeutics. Recently, we have developed pH-sensitive carbonate apatite nanoparticles to efficiently carry and transport siRNA across the cell membrane, enabling knockdown of the cyclin B1 gene and consequential induction of apoptosis in synergy with anti-cancer drugs.Methods and results: We report that carbonate apatite-mediated delivery of the siRNAs targeting ABCG2 and ABCB1 gene transcripts in human breast cancer cells which constitutively express both of the transporter genes dose-dependently enhanced chemosensitivity to doxorubicin, paclitaxel and cisplatin, the traditionally used chemotherapeutic agents. Moreover, codelivery of two specific siRNAs targeting ABCB1 and ABCG2 transcripts resulted in a more robust increase of chemosensitivity in the cancer cells, indicating the reversal of ABC transporter-mediated multidrug resistance.Conclusion: The delivery concept of multiple siRNAs against ABC transporter genes is highly promising for preclinical and clinical investigation in reversing the multidrug resistance phenotype of breast cancer.Keywords: carbonate apatite, siRNA, gene expression, transfection, breast cancer, ABC transporter

  9. Alleviation of temperature-sensitive secretion defect of Pseudomonas fluorescens ATP-binding cassette (ABC) transporter, TliDEF, by a change of single amino acid in the ABC protein, TliD.

    Eom, Gyeong Tae; Oh, Joon Young; Park, Ji Hyun; Lim, Hye Jin; Lee, So Jeong; Kim, Eun Young; Choi, Ji-Eun; Jegal, Jonggeon; Song, Bong Keun; Yu, Ju-Hyun; Song, Jae Kwang

    2016-09-01

    An ABC transporter, TliDEF, from Pseudomonas fluorescens SIK W1, mediates the secretion of its cognate lipase, TliA, in a temperature-dependent secretion manner; the TliDEF-mediated secretion of TliA was impossible at the temperatures over 33°C. To isolate a mutant TliDEF capable of secreting TliA at 35°C, the mutagenesis of ABC protein (TliD) was performed. The mutated tliD library where a random point mutation was introduced by error-prone PCR was coexpressed with the wild-type tliE, tliF and tliA in Escherichia coli. Among approximately 10,000 colonies of the tliD library, we selected one colony that formed transparent halo on LB-tributyrin plates at 35°C. At the growth temperature of 35°C, the selected mutant TliD showed 1.75 U/ml of the extracellular lipase activity, while the wild-type TliDEF did not show any detectable lipase activity in the culture supernatant of E. coli. Moreover, the mutant TliD also showed higher level of TliA secretion than the wild-type TliDEF at other culture temperatures, 20°C, 25°C and 30°C. The mutant TliD had a single amino acid change (Ser287Pro) in the predicted transmembrane region in the membrane domain of TliD, implying that the corresponding region of TliD was important for causing the temperature-dependent secretion of TliDEF. These results suggested that the property of ABC transporter could be changed by the change of amino acid in the ABC protein. PMID:27033673

  10. Cell differentiation and infectivity of Leishmania mexicana are inhibited in a strain resistant to an ABC-transporter blocker.

    Silva, N; Camacho, N; Figarella, K; Ponte-Sucre, A

    2004-06-01

    We analysed whether markers of cell differentiation and infectivity differed when compared to the parental sensitive strain [NR(Gs)] in an in vitro selected Leishmania strain [NR(Gr)] resistant to Glibenclamide, an ATP-binding-cassette (ABC)-transporter blocker. The data show that the cell body area was larger in NR(Gr) compared to NR(Gs) and that functional characters associated with an infective metacyclic phenotype, such as resistance to the lytic effect of the alternative complement pathway and expression of the Meta-1 protein, were reduced. The infectivity of NR(Gr) to J774.1 macrophages was also significantly reduced. These results suggest that resistance in Leishmania against Glibenclamide, a general blocker of P-glycoproteins, could produce functional modifications that may be relevant for Leishmania differentiation, infectivity and survival. PMID:15206465

  11. Salinomycin overcomes ABC transporter-mediated multidrug and apoptosis resistance in human leukemia stem cell-like KG-1a cells

    Fuchs, Dominik [Research Group Molecular Neuro-Oncology, German Cancer Research Center, Im Neuenheimer Feld 280, D-69120 Heidelberg (Germany); Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, D-69120 Heidelberg (Germany); Daniel, Volker; Sadeghi, Mahmoud; Opelz, Gerhard [Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, D-69120 Heidelberg (Germany); Naujokat, Cord, E-mail: cord.naujokat@med.uni-heidelberg.de [Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, D-69120 Heidelberg (Germany)

    2010-04-16

    Leukemia stem cells are known to exhibit multidrug resistance by expression of ATP-binding cassette (ABC) transporters which constitute transmembrane proteins capable of exporting a wide variety of chemotherapeutic drugs from the cytosol. We show here that human promyeloblastic leukemia KG-1a cells exposed to the histone deacetylase inhibitor phenylbutyrate resemble many characteristics of leukemia stem cells, including expression of functional ABC transporters such as P-glycoprotein, BCRP and MRP8. Consequently, KG-1a cells display resistance to the induction of apoptosis by various chemotherapeutic drugs. Resistance to apoptosis induction by chemotherapeutic drugs can be reversed by cyclosporine A, which effectively inhibits the activity of P-glycoprotein and BCRP, thus demonstrating ABC transporter-mediated drug resistance in KG-1a cells. However, KG-1a are highly sensitive to apoptosis induction by salinomycin, a polyether ionophore antibiotic that has recently been shown to kill human breast cancer stem cell-like cells and to induce apoptosis in human cancer cells displaying multiple mechanisms of drug and apoptosis resistance. Whereas KG-1a cells can be adapted to proliferate in the presence of apoptosis-inducing concentrations of bortezomib and doxorubicin, salinomycin does not permit long-term adaptation of the cells to apoptosis-inducing concentrations. Thus, salinomycin should be regarded as a novel and effective agent for the elimination of leukemia stem cells and other tumor cells exhibiting ABC transporter-mediated multidrug resistance.

  12. Salinomycin overcomes ABC transporter-mediated multidrug and apoptosis resistance in human leukemia stem cell-like KG-1a cells

    Leukemia stem cells are known to exhibit multidrug resistance by expression of ATP-binding cassette (ABC) transporters which constitute transmembrane proteins capable of exporting a wide variety of chemotherapeutic drugs from the cytosol. We show here that human promyeloblastic leukemia KG-1a cells exposed to the histone deacetylase inhibitor phenylbutyrate resemble many characteristics of leukemia stem cells, including expression of functional ABC transporters such as P-glycoprotein, BCRP and MRP8. Consequently, KG-1a cells display resistance to the induction of apoptosis by various chemotherapeutic drugs. Resistance to apoptosis induction by chemotherapeutic drugs can be reversed by cyclosporine A, which effectively inhibits the activity of P-glycoprotein and BCRP, thus demonstrating ABC transporter-mediated drug resistance in KG-1a cells. However, KG-1a are highly sensitive to apoptosis induction by salinomycin, a polyether ionophore antibiotic that has recently been shown to kill human breast cancer stem cell-like cells and to induce apoptosis in human cancer cells displaying multiple mechanisms of drug and apoptosis resistance. Whereas KG-1a cells can be adapted to proliferate in the presence of apoptosis-inducing concentrations of bortezomib and doxorubicin, salinomycin does not permit long-term adaptation of the cells to apoptosis-inducing concentrations. Thus, salinomycin should be regarded as a novel and effective agent for the elimination of leukemia stem cells and other tumor cells exhibiting ABC transporter-mediated multidrug resistance.

  13. Function of the ABC transporters, P-glycoprotein, multidrug resistance protein and breast cancer resistance protein, in minimal residual disease in acute myeloid leukemia.

    Pol, van der M.A.; Broxterman, H.J.; Pater, JM; Feller, N.; Maas, M.; Weijers, GW; Scheffer, G.L.; Allen, JD; Scheper, R.J.; Loevezijn, van A; Ossenkoppele, G.J.; Schuurhuis, G.J.

    2003-01-01

    BACKGROUND AND OBJECTIVES: Relapse is common in acute myeloid leukemia (AML) because of persistence of minimal residual disease (MRD). ABC-transporters P-glycoprotein (Pgp) and multidrug resistance protein (MRP), are thought to contribute to treatment failure, while it is unknown whether breast canc

  14. Polarized location of SLC and ABC drug transporters in monolayer-cultured human hepatocytes.

    Le Vee, Marc; Jouan, Elodie; Noel, Gregory; Stieger, Bruno; Fardel, Olivier

    2015-08-01

    Human hepatocytes cultured in a monolayer configuration represent a well-established in vitro model in liver toxicology, notably used in drug transporter studies. Polarized status of drug transporters, i.e., their coordinated location at sinusoidal or canalicular membranes, remains however incompletely documented in these cultured hepatocytes. The present study was therefore designed to analyze transporter expression and location in such cells. Most of drug transporters were first shown to be present at notable mRNA levels in monolayer-cultured human hepatocytes. Cultured human hepatocytes, which morphologically exhibited bile canaliculi-like structures, were next demonstrated, through immunofluorescence staining, to express the influx transporters organic anion transporting polypeptide (OATP) 1B1, OATP2B1 and organic cation transporter (OCT) 1 and the efflux transporter multidrug resistance-associated protein (MRP) 3 at their sinusoidal pole. In addition, the efflux transporters P-glycoprotein and MRP2 were detected at the canalicular pole of monolayer-cultured human hepatocytes. Moreover, canalicular secretion of reference substrates for the efflux transporters bile salt export pump, MRP2 and P-glycoprotein as well as sinusoidal drug transporter activities were observed. This polarized and functional expression of drug transporters in monolayer-cultured human hepatocytes highlights the interest of using this human in vitro cell model in xenobiotic transport studies. PMID:25862123

  15. Novel understanding of ABC transporters ABCB1/MDR/P-glycoprotein, ABCC2/MRP2, and ABCG2/BCRP in colorectal pathophysiology

    Andersen, Vibeke; Svenningsen, Katrine; Knudsen, Lina Almind;

    2015-01-01

    transporter proteins, inflammatory bowel disease, ulcerative, colitis, Crohns disease, colorectal cancer, colitis, intestinal inflammation, intestinal carcinogenesis, ABCB1/P-glycoprotein (P-gp/CD243/MDR1), ABCC2/multidrug resistance protein 2 (MRP2) and ABCG2/breast cancer resistance protein (BCRP), Abcb1....../Mdr1a, abcc2/Mrp2, abcg2/Bcrp, knock-out mice, tight junction, membrane lipid function. RESULTS: Recently, human studies reported that changes in the levels of ABC transporters were early events in the adenoma-carcinoma sequence leading to CRC. A link between ABCB1, high fat diet and gut microbes in...... which may additionally affect ABC transporter function through nuclear receptors and transcriptional regulation. Another critical role of ABCB1 was suggested by the finding that ABCB1 expression identifies a subpopulation of pro-inflammatory Th17 cells which were resistant to treatment with...

  16. Global Land Transport Infrastructure Requirements

    NONE

    2013-06-01

    Over the next four decades, global passenger and freight travel is expected to double over 2010 levels. In order to accommodate this growth, it is expected that the world will need to add nearly 25 million paved road lane-kilometres and 335 000 rail track kilometres. In addition, it is expected that between 45 000 square kilometres and 77 000 square kilometres of new parking spaces will be added to accommodate vehicle stock growth. These land transport infrastructure additions, when combined with operations, maintenance and repairs, are expected to cost as much as USD 45 trillion by 2050. This publication reports on the International Energy Agency’s (IEA) analysis of infrastructure requirements to support projected road and rail travel through 2050, using the IEA Mobility Model. It considers land transport infrastructure additions to support travel growth to 2050. It also considers potential savings if countries pursue “avoid and shift” policies: in this scenario, cumulative global land transport infrastructure spending could decrease as much as USD 20 trillion by 2050 over baseline projections.

  17. Purification, crystallization and preliminary X-ray diffraction analysis of the putative ABC transporter ATP-binding protein from Thermotoga maritima

    The putative ABC transporter ATP-binding protein TM0222 from T. maritima was cloned, overproduced, purified and crystallized. A complete MAD diffraction data set has been collected to 2.3 Å resolution. Adenosine triphosphate (ATP) binding cassette transporters (ABC transporters) are ATP hydrolysis-dependent transmembrane transporters. Here, the overproduction, purification and crystallization of the putative ABC transporter ATP-binding protein TM0222 from Thermotoga maritima are reported. The protein was crystallized in the hexagonal space group P6422, with unit-cell parameters a = b = 148.49, c = 106.96 Å, γ = 120.0°. Assuming the presence of two molecules in the asymmetric unit, the calculated VM is 2.84 Å3 Da−1, which corresponds to a solvent content of 56.6%. A three-wavelength MAD data set was collected to 2.3 Å resolution from SeMet-substituted TM0222 crystals. Data sets were collected on the BL38B1 beamline at SPring-8, Japan

  18. ABC transporters P-gp and Bcrp do not limit the brain uptake of the novel antipsychotic and anticonvulsant drug cannabidiol in mice.

    Brzozowska, Natalia; Li, Kong M; Wang, Xiao Suo; Booth, Jessica; Stuart, Jordyn; McGregor, Iain S; Arnold, Jonathon C

    2016-01-01

    Cannabidiol (CBD) is currently being investigated as a novel therapeutic for the treatment of CNS disorders like schizophrenia and epilepsy. ABC transporters such as P-glycoprotein (P-gp) and breast cancer resistance protein (Bcrp) mediate pharmacoresistance in these disorders. P-gp and Bcrp are expressed at the blood brain barrier (BBB) and reduce the brain uptake of substrate drugs including various antipsychotics and anticonvulsants. It is therefore important to assess whether CBD is prone to treatment resistance mediated by P-gp and Bcrp. Moreover, it has become common practice in the drug development of CNS agents to screen against ABC transporters to help isolate lead compounds with optimal pharmacokinetic properties. The current study aimed to assess whether P-gp and Bcrp impacts the brain transport of CBD by comparing CBD tissue concentrations in wild-type (WT) mice versus mice devoid of ABC transporter genes. P-gp knockout (Abcb1a/b (-∕-)), Bcrp knockout (Abcg2 (-∕-)), combined P-gp/Bcrp knockout (Abcb1a/b (-∕-) Abcg2 (-∕-)) and WT mice were injected with CBD, before brain and plasma samples were collected at various time-points. CBD results were compared with the positive control risperidone and 9-hydroxy risperidone, antipsychotic drugs that are established ABC transporter substrates. Brain and plasma concentrations of CBD were not greater in P-gp, Bcrp or P-gp/Bcrp knockout mice than WT mice. In comparison, the brain/plasma concentration ratios of risperidone and 9-hydroxy risperidone were profoundly higher in P-gp knockout mice than WT mice. These results suggest that CBD is not a substrate of P-gp or Bcrp and may be free from the complication of reduced brain uptake by these transporters. Such findings provide favorable evidence for the therapeutic development of CBD in the treatment of various CNS disorders. PMID:27257556

  19. ABC-type transporters and cuticle assembly: Linking function to polarity in epidermis cells

    Panikashvili, David; Aharoni, Asaph

    2008-01-01

    The aerial organs of plants are covered with a cuticle, a continuous layer overlaying the outermost cell walls of the epidermis. The cuticle is composed of two major classes of the lipid biopolymers: cutin and waxes, collectively termed cuticular lipids. Biosynthesis and transport of cuticular lipids occur predominantly in the epidermis cells. In the transport pathway, cuticular lipids are exported from their site of biosynthesis in the ER/plastid to the extracellular space through the plasma...

  20. Classification of a Haemophilus influenzae ABC Transporter HI1470/71 through Its Cognate Molybdate Periplasmic Binding Protein, MolA

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C.; Pinkett, Heather W. (CIT); (NWU)

    2014-10-02

    molA (HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB{sub 2}C{sub 2} (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7 {angstrom} resolution, respectively. The MolA-binding protein binds molybdate and tungstate, but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate-binding protein structurally solved. The {approx}100 {mu}M binding affinity for tungstate and molybdate is significantly lower than observed for the class II ModA molybdate-binding proteins that have nanomolar to low micromolar affinity for molybdate. The presence of two molybdate loci in H. influenzae suggests multiple transport systems for one substrate, with molABC constituting a low-affinity molybdate locus.

  1. Classification of a Haemophilus influenzae ABC transporter HI1470/71 through its cognate molybdate periplasmic binding protein, MolA.

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C; Pinkett, Heather W

    2011-11-01

    molA (HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB(2)C(2) (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7 Å resolution, respectively. The MolA-binding protein binds molybdate and tungstate, but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate-binding protein structurally solved. The ∼100 μM binding affinity for tungstate and molybdate is significantly lower than observed for the class II ModA molybdate-binding proteins that have nanomolar to low micromolar affinity for molybdate. The presence of two molybdate loci in H. influenzae suggests multiple transport systems for one substrate, with molABC constituting a low-affinity molybdate locus. PMID:22078568

  2. The novel ABC transporter ABCH1 is a potential target for RNAi-based insect pest control and resistance management.

    Guo, Zhaojiang; Kang, Shi; Zhu, Xun; Xia, Jixing; Wu, Qingjun; Wang, Shaoli; Xie, Wen; Zhang, Youjun

    2015-01-01

    Insect pests cause serious crop damage and develop high-level resistance to chemical insecticides and Bacillus thuringiensis (Bt) insecticidal Cry toxins. A new promising approach for controlling them and overcoming this resistance is RNA interference (RNAi). The RNAi-based insect control strategy depends on the selection of suitable target genes. In this study, we cloned and characterized a novel ABC transporter gene PxABCH1 in diamondback moth, Plutella xylostella (L.). Phylogenetic analysis showed that PxABCH1 is closely related to ABCA and ABCG subfamily members. Spatial-temporal expression detection revealed that PxABCH1 was expressed in all tissues and developmental stages, and highest expressed in head and male adult. Midgut sequence variation and expression analyses of PxABCH1 in all the susceptible and Bt-resistant P. xylostella strains and the functional analysis by sublethal RNAi demonstrated that Cry1Ac resistance was independent of this gene. Silencing of PxABCH1 by a relatively high dose of dsRNA dramatically reduced its expression and resulted in larval and pupal lethal phenotypes in both susceptible and Cry1Ac-resistant P. xylostella strains. To our knowledge, this study provides the first insight into ABCH1 in lepidopterans and reveals it as an excellent target for RNAi-based insect pest control and resistance management. PMID:26333918

  3. The naphthoquinones, vitamin K3 and its structural analog plumbagin, are substrates of the multidrug resistance-linked ABC drug transporter ABCG2

    Shukla, Suneet; Wu, Chung-Pu; Nandigama, Krishnamachary; Ambudkar, Suresh V.

    2007-01-01

    Vitamin K3 (Menadione; 2-methyl-1,4-naphthoquinone) is a structural precursor of vitamins K1 and K2 which are essential for blood clotting. The naturally occurring structural analog of this vitamin, plumbagin (5-hydroxy-menadione), is known to modulate cellular proliferation, apoptosis, carcinogenesis, and radioresistance. We, here, report that both vitamin K3 and plumbagin are substrates of the multidrug resistance-linked ATP binding cassette (ABC) drug transporter, ABCG2. Vitamin K3 and plu...

  4. The ABC transporter MgAtr4 is a virulence factor of Mycosphaerella graminicola that affects colonization of substomatal cavities in wheat leaves

    Stergiopoulos, I.; Zwiers, L.H.; Waard, de M.A.

    2003-01-01

    The role in virulence of the ATP-binding cassette (ABC) transporters MgAtr1, MgAtr2, MgAtr3, MgAtr4, and MgAtr5 from Mycosphaerella graminicola was analyzed by gene disruption or replacement on seedlings of the susceptible wheat cultivar Obelisk. Disruption strains of MgAtr1 and MgAtr2 and replaceme

  5. Overexpression of patA and patB, Which Encode ABC Transporters, Is Associated with Fluoroquinolone Resistance in Clinical Isolates of Streptococcus pneumoniae▿

    Garvey, Mark I.; Baylay, Alison J.; Wong, Ryan L.; Piddock, Laura J. V.

    2010-01-01

    Fifty-seven clinical isolates of Streptococcus pneumoniae were divided into four groups based on their susceptibilities to the fluoroquinolones ciprofloxacin and norfloxacin and the dyes ethidium bromide and acriflavine. Comparative reverse transcription-PCR was used to determine the level of expression of the genes patA and patB, which encode putative ABC transporters. Overexpression was observed in 14 of the 15 isolates that were resistant to both fluoroquinolones and dyes and in only 3 of ...

  6. The putative ABC transporter encoded by the orf19.4531 plays a role in the sensitivity of Candida albicans cells to azole antifungal drugs.

    Jiang, Linghuo; Xu, Dayong; Chen, Zhen; Cao, Yongbing; Gao, Pinghui; Jiang, Yuanying

    2016-05-01

    ATP-binding cassette (ABC) transporters constitute a large superfamily of integral membrane proteins in prokaryotic and eukaryotic cells. In the human fungal pathogenCandida albicans, there are 28 genes encoding ABC transporters and many of them have not been characterized so far. The orf19.4531 (also known as IPF7530) encodes a putative ABC transporter. In this study, we have demonstrated that disruption of orf19.4531 causesC. albicanscells to become tolerant to azoles, but not to polyene antifungals and terbinafine. Therefore, the protein encoded by orf19.4531 is involved in azole sensitivity and we name it asROA1, the regulator of azole sensitivity 1 gene. Consistently, we show that the expression ofROA1is responsive to treatment of either fluconazole or ketoconazole inC. albicans In addition, through a GFP tagging approach, Roa1 is localized in a small punctuate compartment adjacent to the vacuolar membrane. However,ROA1is not essential for thein vitrofilamentation ofC. albicanscells. PMID:26975389

  7. Reversal of resistance by GF120918 in cell lines expressing the ABC half-transporter, MXR

    de Bruin, M; Miyake, K; Litman, Thomas;

    1999-01-01

    in S1-B1-20 and S1-M1-80 cells when incubated in the presence of GF120918 than when incubated with mitoxantrone alone. Thus, GF120918 appears to fit the paradigm of a multispecific blocker and is able to block rhodamine and mitoxantrone efflux by the newly identified mitoxantrone transporter. Further...

  8. Formation of a Chloride-conducting State in the Maltose ATP-binding Cassette (ABC) Transporter.

    Carlson, Michael L; Bao, Huan; Duong, Franck

    2016-06-01

    ATP-binding cassette transporters use an alternating access mechanism to move substrates across cellular membranes. This mode of transport ensures the selective passage of molecules while preserving membrane impermeability. The crystal structures of MalFGK2, inward- and outward-facing, show that the transporter is sealed against ions and small molecules. It has yet to be determined whether membrane impermeability is maintained when MalFGK2 cycles between these two conformations. Through the use of a mutant that resides in intermediate conformations close to the transition state, we demonstrate that not only is chloride conductance occurring, but also to a degree large enough to compromise cell viability. Introduction of mutations in the periplasmic gate lead to the formation of a channel that is quasi-permanently open. MalFGK2 must therefore stay away from these ion-conducting conformations to preserve the membrane barrier; otherwise, a few mutations that increase access to the ion-conducting states are enough to convert an ATP-binding cassette transporter into a channel. PMID:27059961

  9. The Riboswitch Regulates a Thiamine Pyrophosphate ABC Transporter of the Oral Spirochete Treponema denticola ▿ †

    Bian, Jiang; Shen, Hongwu; Tu, Youbin; Yu, Aiming; Li, Chunhao

    2011-01-01

    Thiamine pyrophosphate (TPP), a biologically active form of thiamine (vitamin B1), is an essential cofactor in all living systems. Microorganisms either synthesize TPP via de novo biosynthesis pathways or uptake exogenous thiamine from the environment via specific transporters. The oral spirochete Treponema denticola is an important pathogen that is associated with human periodontal diseases. It lacks a de novo TPP biosynthesis pathway and needs exogenous TPP for growth, suggesting that it ma...

  10. ABC protein transport of MRI contrast agents in canalicular rat liver plasma vesicles and yeast vacuoles

    The mechanism of excretion into bile of hepatospecific magnetic resonance imaging (MRI) contrast media employed labeled Gd-reagents EOB.DTPA, BOPTA, B 20790 (iopanoate-linked), and B 21690 (glycocholate-linked) for measurement in rat liver canalicular plasma membrane vesicles and yeast vacuoles. The presence of ATP gave threefold greater transport of B 20790 and B 21690 than of EOB.DTPA and BOPTA. In yeast vacuoles the ATP stimulatory effect was eightfold with B 20790 and fivefold greater for B 21690, whereas in YCF1- or YLLO115w-deleted yeast cells the transport was significantly reduced and absent from double mutants, YCF1 and YLLO15w. The transport was similar in wild-type and deletant cells for B 21690; taurocholate gave 85% inhibition. These data suggest that bilary secretion of structurally related MRI agents depend on molecular structure. The findings are suggestive as of possible value for clinical diagnosis of inherited hyperbilirubinemias and other liver disorders

  11. Heteronuclear multidimensional NMR and homology modelling studies of the C-terminal nucleotide-binding domain of the human mitochondrial ABC transporter ABCB6

    Human ATP-binding cassette, sub-family B, member 6 (ABCB6) is a mitochondrial ABC transporter, and presumably contributes to iron homeostasis. Aimed at understanding the structural basis for the conformational changes accompanying the substrate-transportation cycle, we have studied the C-terminal nucleotide-binding domain of ABCB6 (ABCB6-C) in both the nucleotide-free and ADP-bound states by heteronuclear multidimensional NMR and homology modelling. A non-linear sampling scheme was utilised for indirectly acquired 13C and 15N dimensions of all 3D triple-resonance NMR experiments, in order to overcome the instability and the low solubility of ABCB6-C. The backbone resonances for approximately 25% of non-proline residues, which are mostly distributed around the functionally important loops and in the Helical domain, were not observed for nucleotide-free form of ABCB6-C. From the pH, temperature and magnetic field strength dependencies of the resonance intensities, we concluded that this incompleteness in the assignments is mainly due to the exchange between multiple conformations at an intermediate rate on the NMR timescale. These localised conformational dynamics remained in ADP-bound ABCB6-C except for the loops responsible for adenine base and α/β-phosphate binding. These results revealed that the localised dynamic cooperativity, which was recently proposed for a prokaryotic ABC MJ1267, also exists in a higher eukaryotic ABC, and is presumably shared by all members of the ABC family. Since the Helical domain is the putative interface to the transmembrane domain, this cooperativity may explain the coupled functions between domains in the substrate-transportation cycle

  12. Lateral gene transfer of an ABC transporter complex between major constituents of the human gut microbiome

    Meehan Conor J

    2012-11-01

    Full Text Available Abstract Background Several links have been established between the human gut microbiome and conditions such as obesity and inflammatory bowel syndrome. This highlights the importance of understanding what properties of the gut microbiome can affect the health of the human host. Studies have been undertaken to determine the species composition of this microbiome and infer functional profiles associated with such host properties. However, lateral gene transfer (LGT between community members may result in misleading taxonomic attributions for the recipient organisms, thus making species-function links difficult to establish. Results We identified a peptides/nickel transport complex whose components differed in abundance based upon levels of host obesity, and assigned the encoded proteins to members of the microbial community. Each protein was assigned to several distinct taxonomic groups, with moderate levels of agreement observed among different proteins in the complex. Phylogenetic trees of these proteins produced clusters that differed greatly from taxonomic attributions and indicated that habitat-directed LGT of this complex is likely to have occurred, though not always between the same partners. Conclusions These findings demonstrate that certain membrane transport systems may be an important factor within an obese-associated gut microbiome and that such complexes may be acquired several times by different strains of the same species. Additionally, an example of individual proteins from different organisms being transferred into one operon was observed, potentially demonstrating a functional complex despite the donors of the subunits being taxonomically disparate. Our results also highlight the potential impact of habitat-directed LGT on the resident microbiota.

  13. Novel understanding of ABC transporters ABCB1/MDR/P-glycoprotein, ABCC2/MRP2, and ABCG2/BCRP in colorectal pathophysiology

    Andersen, Vibeke; Svenningsen, Katrine; Almind Knudsen, Lina; Hansen, Axel Kornerup; Holmskov, Uffe; Stensballe, Allan; Vogel, Ulla

    2015-01-01

    transporter proteins, inflammatory bowel disease, ulcerative, colitis, Crohns disease, colorectal cancer, colitis, intestinal inflammation, intestinal carcinogenesis, ABCB1/P-glycoprotein (P-gp/CD243/MDR1), ABCC2/multidrug resistance protein 2 (MRP2) and ABCG2/breast cancer resistance protein (BCRP), Abcb1....../Mdr1a, abcc2/Mrp2, abcg2/Bcrp, knock-out mice, tight junction, membrane lipid function. RESULTS: Recently, human studies reported that changes in the levels of ABC transporters were early events in the adenoma-carcinoma sequence leading to CRC. A link between ABCB1, high fat diet and gut microbes in...

  14. Membrane topology and functional importance of the periplasmic region of ABC transporter LolCDE.

    Yasuda, Masaki; Iguchi-Yokoyama, Asako; Matsuyama, Shin-ichi; Tokuda, Hajime; Narita, Shin-ichiro

    2009-10-01

    The LolCDE complex is an ATP-binding cassette transporter that mediates the release of newly synthesized lipoproteins from the cytoplasmic membrane of gram-negative bacteria, which results in the initiation of outer-membrane sorting of lipoproteins through the Lol pathway. LolCDE is composed of one copy each of membrane subunits LolC and LolE, and two copies of nucleotide-binding subunit LolD. In this study, we examined the membrane topology of LolC and LolE by PhoA fusion analysis. Both LolC and LolE were found to have four transmembrane segments with a large periplasmic loop exposed to the periplasm. Despite similarities in sequence and topology, the accessibility of a sulfhydryl reagent to Cys introduced into the periplasmic loop suggested that the structure of the periplasmic region differs between LolC and LolE. Inhibition of the release of lipoproteins by the sulfhydryl reagent supported a previous proposal that LolC and LolE have distinct functions. PMID:19809197

  15. Subtle Structural Differences Trigger Inhibitory Activity of Propafenone Analogues at the Two Polyspecific ABC Transporters: P-Glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP).

    Schwarz, Theresa; Montanari, Floriane; Cseke, Anna; Wlcek, Katrin; Visvader, Lene; Palme, Sarah; Chiba, Peter; Kuchler, Karl; Urban, Ernst; Ecker, Gerhard F

    2016-06-20

    The transmembrane ABC transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are widely recognized for their role in cancer multidrug resistance and absorption and distribution of compounds. Furthermore, they are linked to drug-drug interactions and toxicity. Nevertheless, due to their polyspecificity, a molecular understanding of the ligand-transporter interaction, which allows designing of both selective and dual inhibitors, is still in its infancy. This study comprises a combined approach of synthesis, in silico prediction, and in vitro testing to identify molecular features triggering transporter selectivity. Synthesis and testing of a series of 15 propafenone analogues with varied rigidity and basicity of substituents provide first trends for selective and dual inhibitors. Results indicate that both the flexibility of the substituent at the nitrogen atom, as well as the basicity of the nitrogen atom, trigger transporter selectivity. Furthermore, inhibitory activity of compounds at P-gp seems to be much more influenced by logP than those at BCRP. Exploiting these differences further should thus allow designing specific inhibitors for these two polyspecific ABC-transporters. PMID:26970257

  16. Combined phylogeny and neighborhood analysis of the evolution of the ABC transporters conferring multiple drug resistance in hemiascomycete yeasts

    Goffeau André

    2009-10-01

    Full Text Available Abstract Background Pleiotropic Drug Resistant transporters (PDR are members of the ATP-Binding Cassette (ABC subfamily which export antifungals and other xenobiotics in fungi and plants. This subfamily of transmembrane transporters has nine known members in Saccharomyces cerevisiae. We have analyzed the complex evolution of the pleiotropic drug resistance proteins (Pdrp subfamily where gene duplications and deletions occur independently in individual genomes. This study was carried out on 62 Pdrp from nine hemiascomycetous species, seven of which span 6 of the 14 clades of the Saccharomyces complex while the two others species, Debaryomyces hansenii and Yarrowia lipolytica, are further apart from an evolutive point of view. Results Combined phylogenetic and neighborhood analyses enabled us to identify five Pdrp clusters in the Saccharomyces complex. Three of them comprise orthologs of the Pdrp sensu stricto, Pdr5p, Pdr10p, Pdr12p, Pdr15p, Snq2p and YNR070wp. The evolutive pathway of the orthologs of Snq2 and YNR070w is particularly complex due to a tandem gene array in Eremothecium gossypii, Kluyveromyces lactis and Saccharomyces (Lachancea kluyveri. This pathway and different cases of duplications and deletions were clarified by using a neighborhood analysis based on synteny. For the two distant species, Yarrowia lipolytica and Debaryomyces hansenii, no neighborhood evidence is available for these clusters and many homologs of Pdr5 and Pdr15 are phylogenetically assigned to species-based clusters. Two other clusters comprise the orthologs of the sensu lato Pdrp, Aus1p/Pdr11p and YOL075cp respectively. The evolutionary pathway of these clusters is simpler. Nevertheless, orthologs of these genes are missing in some species. Conclusion Numerous duplications were traced among the Hemiascomycetous Pdrp studied. The role of the Whole Genome Duplication (WGD is sorted out and our analyses confirm the common ancestrality of Pdr5p and Pdr15p. A tandem

  17. Requirements for transporting the TMI-2 core

    This report summarizes the requirements associated with the actual transport of core debris from the damaged Three Mile Island Unit 2 reactor to the Idaho National Engineering Laboratory. The requirements are discussed under six headings: accountability, security, quality, safety, environment, and transportation

  18. Classification of a Haemophilus influenzae ABC transporter HI1470/71 through its cognate molybdate periplasmic binding protein, MolA

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C.; Pinkett, Heather W.

    2011-01-01

    molA(HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB2C2 (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7-Å resolution, respectively. The MolA binding protein binds molybdate and tungstate but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate binding protein structurally solved. The ~100 μM binding affinity...

  19. The cystic-fibrosis-associated ΔF508 mutation confers post-transcriptional destabilization on the C. elegans ABC transporter PGP-3

    Liping He

    2012-11-01

    Membrane proteins make up ∼30% of the proteome. During the early stages of maturation, this class of proteins can experience localized misfolding in distinct cellular compartments, such as the cytoplasm, endoplasmic reticulum (ER lumen and ER membrane. ER quality control (ERQC mechanisms monitor folding and determine whether a membrane protein is appropriately folded or is misfolded and warrants degradation. ERQC plays crucial roles in human diseases, such as cystic fibrosis, in which deletion of a single amino acid (F508 results in the misfolding and degradation of the cystic fibrosis transmembrane conductance regulator (CFTR Cl– channel. We introduced the ΔF508 mutation into Caenorhabditis elegans PGP-3, a 12-transmembrane ABC transporter with 15% identity to CFTR. When expressed in intestinal epithelial cells, PGP-3wt was stable and efficiently trafficked to the apical plasma membrane through a COPII-dependent mechanism. However, PGP-3ΔF508 was post-transcriptionally destabilized, resulting in reduced total and apical membrane protein levels. Genetic or physiological activation of the osmotic stress response pathway, which causes accumulation of the chemical chaperone glycerol, stabilized PGP-3ΔF508. Efficient degradation of PGP-3ΔF508 required the function of several C. elegans ER-associated degradation (ERAD homologs, suggesting that destabilization occurs through an ERAD-type mechanism. Our studies show that the ΔF508 mutation causes post-transcriptional destabilization and degradation of PGP-3 in C. elegans epithelial cells. This model, combined with the power of C. elegans genetics, provides a new opportunity to genetically dissect metazoan ERQC.

  20. A role for tungsten in the biology of Campylobacter jejuni: tungstate stimulates formate dehydrogenase activity and is transported via an ultra-high affinity ABC system distinct from the molybdate transporter.

    Smart, Jonathan P; Cliff, Matthew J; Kelly, David J

    2009-11-01

    The food-borne pathogen Campylobacter jejuni possesses no known tungstoenzymes, yet encodes two ABC transporters (Cj0300-0303 and Cj1538-1540) homologous to bacterial molybdate (ModABC) uptake systems and the tungstate transporter (TupABC) of Eubacterium acidaminophilum respectively. The actual substrates and physiological role of these transporters were investigated. Tryptophan fluorescence spectroscopy and isothermal titration calorimetry of the purified periplasmic binding proteins of each system revealed that while Cj0303 is unable to discriminate between molybdate and tungstate (K(D) values for both ligands of 4-8 nM), Cj1540 binds tungstate with a K(D) of 1.0 +/- 0.2 pM; 50 000-fold more tightly than molybdate. Induction-coupled plasma mass spectroscopy of single and double mutants showed that this large difference in affinity is reflected in a lower cellular tungsten content in a cj1540 (tupA) mutant compared with a cj0303c (modA) mutant. Surprisingly, formate dehydrogenase (FDH) activity was decreased approximately 50% in the tupA strain, and supplementation of the growth medium with tungstate significantly increased FDH activity in the wild type, while inhibiting known molybdoenzymes. Our data suggest that C. jejuni possesses a specific, ultra-high affinity tungstate transporter that supplies tungsten for incorporation into FDH. Furthermore, possession of two MoeA paralogues may explain the formation of both molybdopterin and tungstopterin in this bacterium. PMID:19818021

  1. The quorum-sensing molecule farnesol is a modulator of drug efflux mediated by ABC multidrug transporters and synergizes with drugs in Candida albicans.

    Sharma, Monika; Prasad, Rajendra

    2011-10-01

    Overexpression of the CaCDR1-encoded multidrug efflux pump protein CaCdr1p (Candida drug resistance protein 1), belonging to the ATP binding cassette (ABC) superfamily of transporters, is one of the most prominent contributors of multidrug resistance (MDR) in Candida albicans. Thus, blocking or modulating the function of the drug efflux pumps represents an attractive approach in combating MDR. In the present study, we provide first evidence that the quorum-sensing molecule farnesol (FAR) is a specific modulator of efflux mediated by ABC multidrug transporters, such as CaCdr1p and CaCdr2p of C. albicans and ScPdr5p of Saccharomyces cerevisiae. Interestingly, FAR did not modulate the efflux mediated by the multidrug extrusion pump protein CaMdr1p, belonging to the major facilitator superfamily (MFS). Kinetic data revealed that FAR competitively inhibited rhodamine 6G efflux in CaCdr1p-overexpressing cells, with a simultaneous increase in an apparent K(m) without affecting the V(max) values and the ATPase activity. We also observed that when used in combination, FAR at a nontoxic concentration synergized with the drugs at their respective nonlethal concentrations, as was evident from their resistant clinical isolates of C. albicans. Our biochemical experiments revealed that the synergistic interaction of FAR with the drugs led to reactive oxygen species accumulation, which triggered early apoptosis, and that both could be partly reversed by the addition of an antioxidant. Collectively, FAR modulates drug extrusion mediated exclusively by ABC proteins and is synergistic to fluconazole (FLC), ketoconazole (KTC), miconazole (MCZ), and amphotericin (AMB). PMID:21768514

  2. Synthesis and characterization of a BODIPY conjugate of the BCR-ABL kinase inhibitor Tasigna® (Nilotinib): Evidence for transport of Tasigna® and its fluorescent derivative by ABC drug transporters

    Shukla, Suneet; Skoumbourdis, Amanda P.; Walsh, Martin J.; Hartz, Anika M. S.; Fung, King Leung; Wu, Chung-pu; Gottesman, Michael M.; Bauer, Björn; Thomas, Craig J.; Suresh V Ambudkar

    2011-01-01

    Tasigna® (Nilotinib) is a recently approved BCR-ABL kinase inhibitor by the Food and Drug Administration, which is indicated for the treatment of drug-resistant chronic myelogenous leukemia (CML). The efflux of tyrosine kinase inhibitors by ATP-binding cassette (ABC) drug transporters, which actively pump these drugs out of cells utilizing ATP as an energy source, has been linked to the development of drug resistance in CML patients. We report here synthesis and characterization of a fluoresc...

  3. Two electrical potential-dependent steps are required for transport by the Escherichia coli Tat machinery.

    Bageshwar, Umesh K; Musser, Siegfried M

    2007-10-01

    The twin-arginine translocation (Tat) pathway in Escherichia coli transports fully folded and assembled proteins across the energy-transducing periplasmic membrane. In chloroplasts, Tat transport requires energy input only from the proton motive force. To elucidate the mechanism and energetics of bacterial Tat protein transport, we developed an efficient in vitro transport assay using TatABC-enriched inverted membrane vesicles and the physiological precursor pre-SufI. We report transport efficiencies of 60-80% for nanomolar pre-SufI concentrations. Dissipation of the pH gradient does not reduce pre-SufI transport efficiency. Instead, pre-SufI transport requires at least two electrical potential (Deltapsi)-dependent steps that differ in both the duration and minimum magnitude of the required Deltapsi. The data are consistent with a model in which a substantial Deltapsi of short duration is required for an early transport step, and in which a small Deltapsi of long duration is necessary to drive a later transport step. PMID:17908913

  4. Quantitative Analysis of the Relative Transcript Levels of ABC Transporter Atr Genes in Aspergillus nidulans by Real-Time Reverse Transcription-PCR Assay

    Pizeta Semighini, Camile; Marins, Mozart; Goldman, Maria Helena S.; Goldman, Gustavo Henrique

    2002-01-01

    The development of assays for quantitative analysis of the relative transcript levels of ABC transporter genes by real-time reverse transcription-PCR (RT-PCR) might provide important information about multidrug resistance in filamentous fungi. Here, we evaluate the potential of real-time RT-PCR to quantify the relative transcript levels of ABC transporter Atr genes from Aspergillus nidulans. The AtrA to AtrD genes showed different and higher levels in the presence of structurally unrelated drugs, such as camptothecin, imazalil, itraconazole, hygromycin, and 4-nitroquinoline oxide. We also verified the relative transcript levels of the Atr genes in the A. nidulans imazalil-resistant mutants. These genes displayed a very complex pattern in different ima genetic backgrounds. The imaB mutant has higher basal transcript levels of AtrB and -D than those of the wild-type strain. The levels of these two genes are comparable when the imaB mutant is grown in the presence and absence of imazalil. The imaC, -D, and -H mutants have higher basal levels of AtrA than that of the wild type. The same behavior is observed for the relative transcript levels of AtrB in the imaG mutant background. PMID:11872487

  5. The Haemophilus influenzae dprABC genes constitute a competence-inducible operon that requires the product of the tfoX (sxy) gene for transcriptional activation.

    Karudapuram, S; Barcak, G J

    1997-01-01

    We previously showed that dprA is required for efficient processing of linear DNA during cellular transformation in Haemophilus influenzae. In this study the transcriptional regulation of dprA and two downstream genes, dprB and dprC, is examined. We demonstrate by Northern blot analysis that the dprABC genes are transcriptionally coregulated and competence inducible. We used primer extension analysis to map the transcriptional start site of dprA and of rec-2, another transformation gene invol...

  6. Detergent screening and purification of the human liver ABC transporters BSEP (ABCB11 and MDR3 (ABCB4 expressed in the yeast Pichia pastoris.

    Philipp Ellinger

    Full Text Available The human liver ATP-binding cassette (ABC transporters bile salt export pump (BSEP/ABCB11 and the multidrug resistance protein 3 (MDR3/ABCB4 fulfill the translocation of bile salts and phosphatidylcholine across the apical membrane of hepatocytes. In concert with ABCG5/G8, these two transporters are responsible for the formation of bile and mutations within these transporters can lead to severe hereditary diseases. In this study, we report the heterologous overexpression and purification of human BSEP and MDR3 as well as the expression of the corresponding C-terminal GFP-fusion proteins in the yeast Pichia pastoris. Confocal laser scanning microscopy revealed that BSEP-GFP and MDR3-GFP are localized in the plasma membrane of P. pastoris. Furthermore, we demonstrate the first purification of human BSEP and MDR3 yielding ∼1 mg and ∼6 mg per 100 g of wet cell weight, respectively. By screening over 100 detergents using a dot blot technique, we found that only zwitterionic, lipid-like detergents such as Fos-cholines or Cyclofos were able to extract both transporters in sufficient amounts for subsequent functional analysis. For MDR3, fluorescence-detection size exclusion chromatography (FSEC screens revealed that increasing the acyl chain length of Fos-Cholines improved monodispersity. BSEP purified in n-dodecyl-β-D-maltoside or Cymal-5 after solubilization with Fos-choline 16 from P. pastoris membranes showed binding to ATP-agarose. Furthermore, detergent-solubilized and purified MDR3 showed a substrate-inducible ATPase activity upon addition of phosphatidylcholine lipids. These results form the basis for further biochemical analysis of human BSEP and MDR3 to elucidate the function of these clinically relevant ABC transporters.

  7. An ABC Transporter Mutation Alters Root Exudation of Phytochemicals That Provoke an Overhaul of Natural Soil Microbiota1[C][W][OA

    Badri, Dayakar V.; Quintana, Naira; El Kassis, Elie G.; Kim, Hye Kyong; Choi, Young Hae; Sugiyama, Akifumi; Verpoorte, Robert; Martinoia, Enrico; Manter, Daniel K.; Vivanco, Jorge M.

    2009-01-01

    Root exudates influence the surrounding soil microbial community, and recent evidence demonstrates the involvement of ATP-binding cassette (ABC) transporters in root secretion of phytochemicals. In this study, we examined effects of seven Arabidopsis (Arabidopsis thaliana) ABC transporter mutants on the microbial community in native soils. After two generations, only the Arabidopsis abcg30 (Atpdr2) mutant had significantly altered both the fungal and bacterial communities compared with the wild type using automated ribosomal intergenic spacer analysis. Similarly, root exudate profiles differed between the mutants; however, the largest variance from the wild type (Columbia-0) was observed in abcg30, which showed increased phenolics and decreased sugars. In support of this biochemical observation, whole-genome expression analyses of abcg30 roots revealed that some genes involved in biosynthesis and transport of secondary metabolites were up-regulated, while some sugar transporters were down-regulated compared with genome expression in wild-type roots. Microbial taxa associated with Columbia-0 and abcg30 cultured soils determined by pyrosequencing revealed that exudates from abcg30 cultivated a microbial community with a relatively greater abundance of potentially beneficial bacteria (i.e. plant-growth-promoting rhizobacteria and nitrogen fixers) and were specifically enriched in bacteria involved in heavy metal remediation. In summary, we report how a single gene mutation from a functional plant mutant influences the surrounding community of soil organisms, showing that genes are not only important for intrinsic plant physiology but also for the interactions with the surrounding community of organisms as well. PMID:19854857

  8. A burst of ABC genes in the genome of the polyphagous spider mite Tetranychus urticae

    Dermauw, W.; Osborne, E.J.; Clark, R.M.; Grbić, M.; Tirry, L.; Van Leeuwen, T

    2013-01-01

    Background: The ABC (ATP-binding cassette) gene superfamily is widespread across all living species. The majority of ABC genes encode ABC transporters, which are membrane-spanning proteins capable of transferring substrates across biological membranes by hydrolyzing ATP. Although ABC transporters have often been associated with resistance to drugs and toxic compounds, within the Arthropoda ABC gene families have only been characterized in detail in several insects and a crustacean. In this st...

  9. Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation

    Teixeira Miguel C

    2012-07-01

    Full Text Available Abstract Background The understanding of the molecular basis of yeast tolerance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. A set of 21 genes encoding multidrug transporters from the ATP-Binding Cassette (ABC Superfamily and Major Facilitator Superfamily (MFS in S. cerevisiae were scrutinized for a role in ethanol stress resistance. Results A yeast multidrug resistance ABC transporter encoded by the PDR18 gene, proposed to play a role in the incorporation of ergosterol in the yeast plasma membrane, was found to confer resistance to growth inhibitory concentrations of ethanol. PDR18 expression was seen to contribute to decreased 3 H-ethanol intracellular concentrations and decreased plasma membrane permeabilization of yeast cells challenged with inhibitory ethanol concentrations. Given the increased tolerance to ethanol of cells expressing PDR18, the final concentration of ethanol produced during high gravity alcoholic fermentation by yeast cells devoid of PDR18 was lower than the final ethanol concentration produced by the corresponding parental strain. Moreover, an engineered yeast strain in which the PDR18 promoter was replaced in the genome by the stronger PDR5 promoter, leading to increased PDR18 mRNA levels during alcoholic fermentation, was able to attain a 6 % higher ethanol concentration and a 17 % higher ethanol production yield than the parental strain. The improved fermentative performance of yeast cells over-expressing PDR18 was found to correlate with their increased ethanol tolerance and ability to restrain plasma membrane permeabilization induced throughout high gravity fermentation. Conclusions PDR18 gene over-expression increases yeast ethanol tolerance and fermentation performance leading to the production of highly inhibitory concentrations of ethanol. PDR18 overexpression in industrial yeast strains appears to be a promising approach to

  10. Identification of the minimal region in lipase ABC transporter recognition domain of Pseudomonas fluorescens for secretion and fluorescence of green fluorescent protein

    Park Yeonwoo

    2012-05-01

    Full Text Available Abstract Background TliA is a thermostable lipase secreted by the type 1 secretion system (T1SS of Pseudomonas fluorescens. The secretion is promoted by its secretion/chaperone domain located near the C-terminus, which is composed mainly of four Repeat-in-Toxin (RTX repeats. In order to identify the minimal region of TliA responsible for its secretion, five different copies of the secretion/chaperone domain, each involving truncated N-terminal residues and a common C-terminus, were acquired and named as lipase ABC transporter recognition domains (LARDs. Each LARD was fused to epidermal growth factor (EGF or green fluorescent protein (GFP, and the secretion of EGF-LARD or GFP-LARD fusion proteins was assessed in Escherichia coli with ABC transporter. Results Among the fusion proteins, GFP or EGF with 105-residue LARD3 was most efficiently secreted. In addition, GFP-LARD3 emitted wild type GFP fluorescence. Structurally, LARD3 had the 4 RTX repeats exposed at the N-terminus, while other LARDs had additional residues prior to them or missed some of the RTX repeats. LARD3 was both necessary and sufficient for efficient secretion and maintenance of GFP fluorescence in E. coli, which was also confirmed in P. fluorescens and P. fluorescens ▵tliA, a knock-out mutant of tliA. Conclusion LARD3 was a potent secretion signal in T1SS for its fusion flanking RTX motif, which enhanced secretion and preserved the fluorescence of GFP. LARD3-mediated secretion in E. coli or P. fluorescens will enable the development of enhanced protein manufacturing factory and recombinant microbe secreting protein of interest in situ.

  11. USMC tactical motor transport lift requirements model

    Allen, Scott Andrew

    1995-01-01

    This thesis concentrates on developing a spreadable model that can be used by Marine logisticians in computing sustainment requirements and the resulting tactical motor transport lift requirements necessary to keep a notional sized maneuver element supported on a daily basis in the Marine Corps projected maneuver warfare environment. Sustainment computations are limited to resupplying the maneuver element with food, water, fuel, and ammunition. Using an "add-in" simulation package the plannin...

  12. Transport package maintenance requirements and operations

    The NuPac 125-B rail cask, which transports the damaged core debris from Three Mile Island Unit 2 (TMI-2) to the Idaho National Engineering Laboratory (INEL), is the only new spent-fuel rail shipping cask to be licensed in the United States within the last decade. EG ampersand G Idaho, Inc. (EG ampersand G), acting on behalf of the US Department of Energy, is responsible for ensuring that those new casks and rail cars are properly maintained per regulatory requirements. Both the casks and rail cars have comprehensive in-service inspection and preventive maintenance programs, which are more involved than implied by the requirements. The TMI-2 shipping campaign is the most ambitious spent-fuel transport activity being conducted in the nuclear industry. The experience gained in this campaign, as it relates to maintenance of a transport system, should be of interest and have direct applicability to similar shipping activities planned in the years ahead

  13. Effects of in vitro exposure to ivermectin and levamisole on the expression patterns of ABC transporters in Haemonchus contortus larvae

    Ali Raza

    2016-08-01

    Full Text Available This study investigated the interaction of ATP binding cassette (ABC transport proteins with ivermectin (IVM and levamisole (LEV in larvae of susceptible and resistant isolates of Haemonchus contortus in vitro by measuring transcription patterns following exposure to these anthelmintics. Furthermore, we studied the consequences of drug exposure by measuring the sensitivity of L3 to subsequent exposure to higher drug concentrations using larval migration assays. The most highly transcribed transporter genes in both susceptible and resistant L3 were pgp-9.3, abcf-1, mrp-5, abcf-2, pgp-3, and pgp-10. The resistant isolate showed significantly higher transcription of pgp-1, pgp-9.1 and pgp-9.2 compared to the susceptible isolate. Five P-gp genes and the haf-6 gene showed significantly higher transcription (up to 12.6-fold after 3 h exposure to IVM in the resistant isolate. Similarly, five P-gp genes, haf-6 and abcf-1 were transcribed at significantly higher levels (up to 10.3-fold following 3 h exposure to LEV in this isolate. On the other hand, there were no significant changes in transcriptional patterns of all transporter genes in the susceptible isolate following 3 and 6 h exposure to IVM or LEV. In contrast to these isolate-specific transcription changes, both isolates showed an increase in R-123 efflux following exposure to the drugs, suggesting that the drugs stimulated activity of existing transporter proteins in both isolates. Exposure of resistant larvae to IVM or LEV resulted, in some instances, in an increase in the proportion of the population able to migrate at the highest IVM concentrations in subsequent migration assays. The significant increase in transcription of some ABC transporter genes following 3 h exposure to both IVM and LEV in the resistant isolate only, suggests that an ability to rapidly upregulate protective pathways in response to drugs may be a component of the resistance displayed by this isolate.

  14. The Nucleotide-Free State of the Multidrug Resistance ABC Transporter LmrA: Sulfhydryl Cross-Linking Supports a Constant Contact, Head-to-Tail Configuration of the Nucleotide-Binding Domains.

    Peter M Jones

    Full Text Available ABC transporters are integral membrane pumps that are responsible for the import or export of a diverse range of molecules across cell membranes. ABC transporters have been implicated in many phenomena of medical importance, including cystic fibrosis and multidrug resistance in humans. The molecular architecture of ABC transporters comprises two transmembrane domains and two ATP-binding cassettes, or nucleotide-binding domains (NBDs, which are highly conserved and contain motifs that are crucial to ATP binding and hydrolysis. Despite the improved clarity of recent structural, biophysical, and biochemical data, the seemingly simple process of ATP binding and hydrolysis remains controversial, with a major unresolved issue being whether the NBD protomers separate during the catalytic cycle. Here chemical cross-linking data is presented for the bacterial ABC multidrug resistance (MDR transporter LmrA. These indicate that in the absence of nucleotide or substrate, the NBDs come into contact to a significant extent, even at 4°C, where ATPase activity is abrogated. The data are clearly not in accord with an inward-closed conformation akin to that observed in a crystal structure of V. cholerae MsbA. Rather, they suggest a head-to-tail configuration 'sandwich' dimer similar to that observed in crystal structures of nucleotide-bound ABC NBDs. We argue the data are more readily reconciled with the notion that the NBDs are in proximity while undergoing intra-domain motions, than with an NBD 'Switch' mechanism in which the NBD monomers separate in between ATP hydrolysis cycles.

  15. Industrial Requirements for Thermodynamics and Transport Properties

    Hendriks, Eric; Kontogeorgis, Georgios; Dohrn, Ralf;

    2010-01-01

    This work reports the results of an investigation on industrial requirements for thermodynamic and transport properties carried out by the Working Party on Thermodynamic and Transport properties (http://www.wp-ttp.dk/) of the European Federation of Chemical Engineering, EFCE (http...... collaboration. The main results are as follows. There is (still) an acute need for accurate, reliable, and thermodynamically consistent experimental data. Quality is more important than quantity. Similarly, there is a great need for reliable predictive, rather than correlative, models covering a wide range of...... compositions, temperatures, and pressures and capable of predicting primary (phase equilibrium) and secondary (enthalpy, heat capacity, etc.) properties. It is clear that the ideal of a single model covering all requirements is not achievable, but there is a consensus that this ideal should still provide the...

  16. The Half-Size ABC Transporter FOLDED PETALS 2/ABCG13 Is Involved in Petal Elongation through Narrow Spaces in Arabidopsis thaliana Floral Buds

    Seiji Takeda

    2014-08-01

    Full Text Available Flowers are vital for attracting pollinators to plants and in horticulture for humans. Petal morphogenesis is a central process of floral development. Petal development can be divided into three main processes: the establishment of organ identity in a concentric pattern, primordia initiation at fixed positions within a whorl, and morphogenesis, which includes petal elongation through the narrow spaces within the bud. Here, we show that the FOLDED PETALS 2 (FOP2 gene, encoding a member of the half-size ATP binding cassette (ABC transporter family ABCG13, is involved in straight elongation of petals in Arabidopsis thaliana. In fop2 mutants, flowers open with folded petals, instead of straight-elongated ones found in the wild type. The epicuticular nanoridge structures are absent in many abaxial epidermal cells of fop2 petals, and surgical or genetic generation of space in young fop2 buds restores the straight elongation of petals, suggesting that the physical contact of sepals and petals causes the petal folding. Similar petal folding has been reported in the fop1 mutant, and the petals of fop2 fop1 double mutants resemble those of both the fop1 and fop2 single mutants, although the epidermal structure and permeability of the petal surface is more affected in fop2. Our results suggest that synthesis and transport of cutin or wax in growing petals play an important role for their smooth elongation through the narrow spaces of floral buds.

  17. ABC Kinga kauplused = ABC King shoe stores

    2011-01-01

    Tallinnas Kristiine keskuses, Tartu Kaubamajas ja Pärnus Port Artur 2 asuvate ABC Kinga kaupluste sisekujundusest. Sisearhitekid Andres Labi ja Janno Roos (Ruumilabor OÜ), loetletud nende ühiselt tehtud töid

  18. ATP and AMP Mutually Influence Their Interaction with the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) at Separate Binding Sites*

    Randak, Christoph O.; Dong, Qian; Ver Heul, Amanda R.; Elcock, Adrian H.; Welsh, Michael J.

    2013-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel in the ATP-binding cassette (ABC) transporter protein family. In the presence of ATP and physiologically relevant concentrations of AMP, CFTR exhibits adenylate kinase activity (ATP + AMP ⇆ 2 ADP). Previous studies suggested that the interaction of nucleotide triphosphate with CFTR at ATP-binding site 2 is required for this activity. Two other ABC proteins, Rad50 and a structural maintenance of chromosome protein, ...

  19. 9 CFR 88.4 - Requirements for transport.

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Requirements for transport. 88.4... TRANSPORTATION OF EQUINES FOR SLAUGHTER § 88.4 Requirements for transport. (a) Prior to the commercial...) Complete and sign an owner-shipper certificate for each equine being transported. The...

  20. 75 FR 49549 - ABC & D Recycling, Inc.-Lease and Operation Exemption-a Line of Railroad in Ware, MA

    2010-08-13

    ... Surface Transportation Board ABC & D Recycling, Inc.--Lease and Operation Exemption--a Line of Railroad in Ware, MA ABC & D Recycling, Inc. (ABC & D), a noncarrier, has filed a verified notice of exemption... operation of this trackage in FD 35356, ABC & D Recycling, Inc.--Lease and Operation Exemption--a Line...

  1. Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines: mRNA analysis and functional radiotracer studies

    Gomes, Celia Maria Freitas [Department of Pathology, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Faculty of Medicine, Institute of Biophysics/Biomathematics, IBILI, 3000-354 Coimbra (Portugal)]. E-mail: cgomes@ibili.uc.pt; van Paassen, Heidi [Department of Pathology, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Romeo, Salvatore [Department of Pathology, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Welling, Mick M. [Department of Radiology, Section of Nuclear Medicine, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Feitsma, R.I.J. [Department of Radiology, Section of Nuclear Medicine, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Abrunhosa, Antero J. [Faculty of Medicine, Institute of Biophysics/Biomathematics, IBILI, 3000-354 Coimbra (Portugal); Botelho, M. Filomena [Faculty of Medicine, Institute of Biophysics/Biomathematics, IBILI, 3000-354 Coimbra (Portugal); Hogendoorn, Pancras C.W. [Department of Pathology, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Pauwels, Ernest [Department of Radiology, Section of Nuclear Medicine, Leiden University Medical Center, 2300 RC Leiden (Netherlands); Cleton-Jansen, Anne Marie [Department of Pathology, Leiden University Medical Center, 2300 RC Leiden (Netherlands)

    2006-10-15

    Drug resistance remains a significant impediment to successful chemotherapy and constitutes a major prognostic factor in osteosarcoma (OS) patients. This study was designed to identify the role and prognostic significance of multidrug-resistance (MDR)-related transporters, such as multidrug resistance protein 1 (MDR1), multidrug-resistance-associated protein (MRP1) and breast-cancer-related protein (BCRP), in OS using cationic lipophilic radiotracers. We evaluated the chemosensitivity of four OS cell lines (Saos-2, 143B, MNNG/HOS and U-2OS) to doxorubicin (DOX), cisplatin (CIS) and methotrexate. The expression of MDR-related transporters was analyzed at mRNA level by quantitative polymerase chain reaction and at functional level by {sup 99m}Tc sestamibi and {sup 99m}Tc tetrofosmin. The effectiveness of MDR modulators [cyclosporin A (CsA) and imatinib] on transporter inhibition and on the reversal of resistance was also assessed. MNNG/HOS and U-2OS cells expressing high levels of MDR1 were highly resistant to DOX and showed reduced accumulation and higher efflux for radiotracers. Although MRP1 was uniformly expressed in all cells, only U-2OS was resistant to CIS. CsA restored sensitivity to DOX and CIS, and enhanced the accumulation and efflux half-life of radiotracers in MDR1-expressing cell lines. The chemosensitivity of OS cells to DOX was strongly dependent on mRNA MDR1 expression and could be circumvented by adding CsA. The kinetic parameters of radiotracers correlated with MDR1 expression levels, hence predicting DOX resistance. We concluded that sensitivity to chemotherapy is strongly dependent on the expression of MDR1 transporter and that radiotracer studies could prove clinically useful in predicting chemotherapy response and in evaluating the efficacy of MDR-reversing agents.

  2. A Putative ABC Transporter Permease Is Necessary for Resistance to Acidified Nitrite and EDTA in Pseudomonas aeruginosa under Aerobic and Anaerobic Planktonic and Biofilm Conditions

    McDaniel, Cameron; Su, Shengchang; Panmanee, Warunya; Lau, Gee W.; Browne, Tristan; Cox, Kevin; Paul, Andrew T.; Ko, Seung-Hyun B.; Mortensen, Joel E.; Lam, Joseph S.; Muruve, Daniel A.; Hassett, Daniel J.

    2016-01-01

    Pseudomonas aeruginosa (PA) is an important airway pathogen of cystic fibrosis and chronic obstructive disease patients. Multiply drug resistant PA is becoming increasing prevalent and new strategies are needed to combat such insidious organisms. We have previously shown that a mucoid, mucA22 mutant PA is exquisitely sensitive to acidified nitrite (A-NO2−, pH 6.5) at concentrations that are well tolerated in humans. Here, we used a transposon mutagenesis approach to identify PA mutants that are hypersensitive to A-NO2−. Among greater than 10,000 mutants screened, we focused on PA4455, in which the transposon was found to disrupt the production of a putative cytoplasmic membrane-spanning ABC transporter permease. The PA4455 mutant was not only highly sensitive to A-NO2−, but also the membrane perturbing agent, EDTA and the antibiotics doxycycline, tigecycline, colistin, and chloramphenicol, respectively. Treatment of bacteria with A-NO2− plus EDTA, however, had the most dramatic and synergistic effect, with virtually all bacteria killed by 10 mM A-NO2−, and EDTA (1 mM, aerobic, anaerobic). Most importantly, the PA4455 mutant was also sensitive to A-NO2− in biofilms. A-NO2− sensitivity and an anaerobic growth defect was also noted in two mutants (rmlC and wbpM) that are defective in B-band LPS synthesis, potentially indicating a membrane defect in the PA4455 mutant. Finally, this study describes a gene, PA4455, that when mutated, allows for dramatic sensitivity to the potential therapeutic agent, A-NO2− as well as EDTA. Furthermore, the synergy between the two compounds could offer future benefits against antibiotic resistant PA strains. PMID:27064218

  3. Know Your Diabetes ABCs

    Full Text Available ... among people with diabetes. Judith Fradkin, M.D.: We call this approach "the ABCs of diabetes." A ... the importance of all three of these and we know that controlling the three can dramatically reduce ...

  4. Know Your Diabetes ABCs

    Full Text Available ... just live to survive. Announcer: People with diabetes should work with their doctors to monitor the ABCs ... my A1C, my blood pressure, my cholesterol, what should it be" "and what can I do to ...

  5. Know Your Diabetes ABCs

    Full Text Available ... Announcer: People with diabetes should work with their doctors to monitor the ABCs of diabetes. Judith Fradkin, ... want patients who have diabetes to ask their doctors, "What is my A1C, my blood pressure, my ...

  6. Know Your Diabetes ABCs

    Full Text Available ... the importance of blood pressure, cholesterol and diabetes management to prevent heart attacks and strokes. Sharon Boykin: ... Knowing your ABCs will help you reduce your risk of heart attack and stroke to live a ...

  7. Know Your Diabetes ABCs

    Full Text Available ... between diabetes and heart disease. Judith Fradkin, M.D.: What most people don't know is that ... stroke among people with diabetes. Judith Fradkin, M.D.: We call this approach "the ABCs of diabetes." ...

  8. Know Your Diabetes ABCs

    Full Text Available ... prevent two of the disease's most serious complications. John Buse, M.D.: As the population ages, we ... of living -- your living conditions will be better. John Buse, M.D.: Knowing your ABCs will help ...

  9. Know Your Diabetes ABCs

    Full Text Available ... As the population ages, we would expect some increase in the proportion of people with diabetes and ... ABC numbers are, you'll be able to help save your own life -- the lower they are, ...

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

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

    2013-02-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SSs). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Threonine kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein, Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and, apparatus assembly and effector export. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signalling pathway that promotes H1-T6SS activity under optimal environmental conditions. PMID:22765374

  11. Transportation system requirements document. Revision 1 DCN01. Supplement

    The original Transportation System Requirements Document described the functions to be performed by and the technical requirements for the Transportation System to transport spent nuclear fuel (SNF) and high-level radioactive waste (HLW) from Purchaser and Producer sites to a Civilian Radioactive Waste Management System (CRWMS) site, and between CRWMS sites. The purpose of that document was to define the system-level requirements. These requirements include design and operations requirements to the extent they impact on the development of the physical segments of Transportation. The document also presented an overall description of Transportation, its functions, its segments, and the requirements allocated to the segments and the system-level interfaces with Transportation. This revision of the document contains only the pages that have been modified

  12. The ABC of cholesterol transport

    Plösch, Torsten

    2004-01-01

    Cholesterol fulfills an indispensable role in mammalian physiology. It is an important constituent of all cell membranes. Furthermore, it is the precursor of steroid hormones, which regulate a variety of physiological functions, and of bile salts, which are necessary for the generation of bile flow

  13. 10 CFR 11.13 - Special requirements for transportation.

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Special requirements for transportation. 11.13 Section 11.13 Energy NUCLEAR REGULATORY COMMISSION CRITERIA AND PROCEDURES FOR DETERMINING ELIGIBILITY FOR... Authorization § 11.13 Special requirements for transportation. (a) All individuals who, after 365 days...

  14. Salt Repository Project transportation system interface requirements: Transportation system/repository receiving facility interface requirements

    This report is a preliminary review of the interface between the transportation system and the repository receiving facility for a nuclear waste mined geologic disposal system in salt. Criteria for generic cask and facility designs are developed. These criteria are derived by examining the interfaces that occur as a result of the operations needed to receive nuclear waste at a repository. These criteria provide the basis for design of a safe, operable, practical nuclear waste receiving facility. The processing functions required to move the shipping unit from the gate into the unloading area and back to the gate for dispatch are described. Criteria for a generic receiving facility are discussed but no specific facility design is presented or evaluated. The criteria are stated in general terms to allow application to a wide variety of cask and facility designs. 9 refs., 6 figs., 4 tabs

  15. Know Your Diabetes ABCs

    Full Text Available ... somebody who's already had a heart attack. Announcer: Research shows that controlling blood sugar, blood pressure and cholesterol are critical in preventing heart disease and stroke among people with diabetes. Judith Fradkin, M.D.: We call this approach "the ABCs of diabetes." A is the A1C, ...

  16. Know Your Diabetes ABCs

    Full Text Available ... are at the same risk for heart disease as somebody who's already had a heart attack. Announcer: Research shows that controlling blood ... We call this approach "the ABCs of diabetes." A is the A1C, a measure of how well ...

  17. Rapportering med ABC modellen

    Wiese, Lars Ole

    2006-01-01

    I denne artikel diskuteres Activity Based Costing (ABC) som rapporterings-teknik for kunde profitabilitet. Der argumenteres for, at indtægtserhvervel-sen inkl. dens direkte omkostningsforbrug bør adskilles fra omkostnings-beskrivelsen til produktfremstilling. Dvs. der gennemføres en aktivitetsba...

  18. The ABC exporter MsbA probed by solid state NMR – challenges and opportunities.

    Kaur, Hundeep; Lakatos, Andrea; Spadaccini, Roberta; Vogel, Ramona; Hoffmann, Christian; Becker-Baldus, Johanna; Ouari, Olivier; Tordo, Paul; Mchaourab, Hassane; Glaubitz, Clemens

    2015-09-01

    ATP binding cassette (ABC) transporters form a superfamily of integral membrane proteins involved in translocation of substrates across the membrane driven by ATP hydrolysis. Despite available crystal structures and extensive biochemical data, many open questions regarding their transport mechanisms remain. Therefore, there is a need to explore spectroscopic techniques such as solid state NMR in order to bridge the gap between structural and mechanistic data. In this study, we investigate the feasibility of using Escherichia coli MsbA as a model ABC transporter for solid state NMR studies. We show that optimised solubilisation and reconstitution procedures enable preparing stable and homogenous protein samples. Depending on the duration of solubilisation, MsbA can be obtained in either an apo- or in a native lipid A bound form. Building onto these optimisations, the first promising MAS-NMR spectra with narrow lines have been recorded. However, further sensitivity improvements are required so that complex NMR experiments can be recorded within a reasonable amount of time. We therefore demonstrate the usability of paramagnetic doping for rapid data acquisition and explore dynamic nuclear polarisation as a method for general signal enhancement. Our results demonstrate that solid state NMR provides an opportunity to address important biological questions related to complex mechanisms of ABC transporters. PMID:25849794

  19. Regulatory requirements for the transport of radioactive materials in Canada

    Canada is a major producer and shipper of radioactive material. Each year more than a million packages are transported in Canada. The safety record with the transport of RAM in Canada has historically been excellent. There have never been any serious injuries, overexposure or fatality or environmental consequences attributable to the radioactive nature of such material being transported or being involved in a transport accident. In Canada, the Canadian Nuclear Safety Commission (CNSC) is the prime agency of the federal government entrusted with regulating all activities related to the use of nuclear energy and nuclear substances including the packaging and transport of nuclear substances. The mission of the CNSC is to regulate the use of nuclear energy and materials to protect health, safety, security of the person and the environment and to respect Canada's international commitments on the peaceful use of nuclear energy. The division of responsibility for the regulation of transport of radioactive material has been split between Transport Canada and the CNSC. The governing Transport Canada's regulations are Transport of Dangerous Goods (TDG) Regulations and the CNSC regulations are Packaging and Transport of Nuclear Substances Regulations (PTNSR). Canada has actively participated in the development of the IAEA regulations for the safe transport of radioactive material since 1960. As an IAEA member state, Canada generally follows the requirements of IAEA regulations with few deviations. The Nuclear Safety and Control Act (NSCA) strongly supports Canada's international obligations to ensure safe packaging, transport, storage and disposal of nuclear substances, prescribed equipment and prescribed information. Prescribed equipment and prescribed information are defined in the CNSC General Nuclear Safety and Control Regulations. This paper presents the current CNSC regulatory requirements and initiatives taken by the CNSC to improve its effectiveness and efficiency

  20. A New Activity-Based Cost (ABC) Mathematical Model

    JIANG Shuo; SONG Lei

    2003-01-01

    Along with the product price competition growing intensely, it is apparently important for reasonably distributing and counting cost. But, in sharing indirect cost, traditional cost accounting unveils the limitations increasingly, especially in authenticity of cost information. And the accounting theory circles and industry circles begin seeking one kind of new accurate cost calculation method, and the activity-based cost (ABC) method emerges as the times require. In this paper, we will build its mathematical model by the basic principle of ABC, and will improve its mathematical model further. We will establish its comparison mathematical model and make the ABC method go a step further to its practical application.

  1. Cadmium-inducible expression of the ABC-type transporter AtABCC3 increases phytochelatin-mediated cadmium tolerance in Arabidopsis

    Brunetti, Patrizia; Zanella, Letizia; De Paolis, Angelo; Di Litta, Davide; Cecchetti, Valentina; Falasca, Giuseppina; Barbieri, Maurizio; Altamura, Maria Maddalena; Costantino, Paolo; Cardarelli, Maura

    2015-01-01

    The heavy metal cadmium (Cd) is a widespread environmental contaminant with harmful effects on living cells. In plants, phytochelatin (PC)-dependent Cd detoxification requires that PC–Cd complexes are transported into vacuoles. Here, it is shown that Arabidopsis thaliana seedlings defective in the ABCC transporter AtABCC3 (abcc3) have an increased sensitivity to different Cd concentrations, and that seedlings overexpressing AtABCC3 (AtABCC3ox) have an increased Cd tolerance. The cellular distribution of Cd was analysed in protoplasts from abcc3 mutants and AtABCC3 overexpressors grown in the presence of Cd, by means of the Cd-specific fluorochromes 5-nitrobenzothiazole coumarin (BTC-5N) and Leadmium™ Green AM dye. This analysis revealed that Cd is mostly localized in the cytosol of abcc3 mutant protoplasts whereas there is an increase in vacuolar Cd in protoplasts from AtABCC3ox plants. Overexpression of AtABCC3 in cad1-3 mutant seedlings defective in PC production and in plants treated with l-buthionine sulphoximine (BSO), an inhibitor of PC biosynthesis, had no effect on Cd tolerance, suggesting that AtABCC3 acts via PCs. In addition, overexpression of AtABCC3 in atabcc1 atabcc2 mutant seedlings defective in the Cd transporters AtABCC1 and AtABCC2 complements the Cd sensitivity of double mutants, but not in the presence of BSO. Accordingly, the level of AtABCC3 transcript in wild type seedlings was lower than that of AtABCC1 and AtABCC2 in the absence of Cd but higher after Cd exposure, and even higher in atabcc1 atabcc2 mutants. The results point to AtABCC3 as a transporter of PC–Cd complexes, and suggest that its activity is regulated by Cd and is co-ordinated with the activity of AtABCC1/AtABCC2. PMID:25900618

  2. Cadmium-inducible expression of the ABC-type transporter AtABCC3 increases phytochelatin-mediated cadmium tolerance in Arabidopsis.

    Brunetti, Patrizia; Zanella, Letizia; De Paolis, Angelo; Di Litta, Davide; Cecchetti, Valentina; Falasca, Giuseppina; Barbieri, Maurizio; Altamura, Maria Maddalena; Costantino, Paolo; Cardarelli, Maura

    2015-07-01

    The heavy metal cadmium (Cd) is a widespread environmental contaminant with harmful effects on living cells. In plants, phytochelatin (PC)-dependent Cd detoxification requires that PC-Cd complexes are transported into vacuoles. Here, it is shown that Arabidopsis thaliana seedlings defective in the ABCC transporter AtABCC3 (abcc3) have an increased sensitivity to different Cd concentrations, and that seedlings overexpressing AtABCC3 (AtABCC3ox) have an increased Cd tolerance. The cellular distribution of Cd was analysed in protoplasts from abcc3 mutants and AtABCC3 overexpressors grown in the presence of Cd, by means of the Cd-specific fluorochromes 5-nitrobenzothiazole coumarin (BTC-5N) and Leadmium™ Green AM dye. This analysis revealed that Cd is mostly localized in the cytosol of abcc3 mutant protoplasts whereas there is an increase in vacuolar Cd in protoplasts from AtABCC3ox plants. Overexpression of AtABCC3 in cad1-3 mutant seedlings defective in PC production and in plants treated with l-buthionine sulphoximine (BSO), an inhibitor of PC biosynthesis, had no effect on Cd tolerance, suggesting that AtABCC3 acts via PCs. In addition, overexpression of AtABCC3 in atabcc1 atabcc2 mutant seedlings defective in the Cd transporters AtABCC1 and AtABCC2 complements the Cd sensitivity of double mutants, but not in the presence of BSO. Accordingly, the level of AtABCC3 transcript in wild type seedlings was lower than that of AtABCC1 and AtABCC2 in the absence of Cd but higher after Cd exposure, and even higher in atabcc1 atabcc2 mutants. The results point to AtABCC3 as a transporter of PC-Cd complexes, and suggest that its activity is regulated by Cd and is co-ordinated with the activity of AtABCC1/AtABCC2. PMID:25900618

  3. The Tangier disease gene product ABC1 controls the cellular apolipoprotein-mediated lipid removal pathway

    Lawn, Richard M.; Wade, David P.; Garvin, Michael R.; Wang, Xingbo; Schwartz, Karen; Porter, J. Gordon; Seilhamer, Jeffrey J.; Ashley M Vaughan; Oram, John F.

    1999-01-01

    The ABC1 transporter was identified as the defect in Tangier disease by a combined strategy of gene expression microarray analysis, genetic mapping, and biochemical studies. Patients with Tangier disease have a defect in cellular cholesterol removal, which results in near zero plasma levels of HDL and in massive tissue deposition of cholesteryl esters. Blocking the expression or activity of ABC1 reduces apolipoprotein-mediated lipid efflux from cultured cells, and increasing expression of ABC...

  4. Flavonoid-mediated inhibition of intestinal ABC transporters may affect the oral bioavailability of drugs, food-borne toxic compounds and bioactive ingredients

    Brand, W.; Schutte, M.E.; Bladeren, van P.J.; Rietjens, I.M.C.M.

    2006-01-01

    The transcellular transport of ingested food ingredients across the intestinal epithelial barrier is an important factor determining bioavailability upon oral intake. This transcellular transport of many chemicals, food ingredients, drugs or toxic compounds over the intestinal epithelium can be high

  5. Domain Interactions in the Yeast ATP Binding Cassette Transporter Ycf1p: Intragenic Suppressor Analysis of Mutations in the Nucleotide Binding Domains

    Falcón-Pérez, Juan M.; Martínez-Burgos, Mónica; Molano, Jesús; Mazón, María J.; Eraso, Pilar

    2001-01-01

    The yeast cadmium factor (Ycf1p) is a vacuolar ATP binding cassette (ABC) transporter required for heavy metal and drug detoxification. Cluster analysis shows that Ycf1p is strongly related to the human multidrug-associated protein (MRP1) and cystic fibrosis transmembrane conductance regulator and therefore may serve as an excellent model for the study of eukaryotic ABC transporter structure and function. Identifying intramolecular interactions in these transporters may help to elucidate ener...

  6. Space Transportation System Availability Requirement and Its Influencing Attributes Relationships

    Rhodes, Russell E.; Adams, Timothy C.; McCleskey, Carey M.

    2008-01-01

    It is important that engineering and management accept the need for an availability requirement that is derived with its influencing attributes. It is the intent of this paper to provide the visibility of relationships of these major attribute drivers (variables) to each other and the resultant system inherent availability. Also important to provide bounds of the variables providing engineering the insight required to control the system's engineering solution, e.g., these influencing attributes become design requirements also. These variables will drive the need to provide integration of similar discipline functions or technology selection to allow control of the total parts count. The relationship of selecting a reliability requirement will place a constraint on parts count to achieve a given availability requirement or if allowed to increase the parts count will drive the system reliability requirement higher. They also provide the understanding for the relationship of mean repair time (or mean down time) to maintainability, e.g., accessibility for repair, and both the mean time between failure, e.g., reliability of hardware and availability. The concerns and importance of achieving a strong availability requirement is driven by the need for affordability, the choice of using the two launch solution for the single space application, or the need to control the spare parts count needed to support the long stay in either orbit or on the surface of the moon. Understanding the requirements before starting the architectural design concept will avoid considerable time and money required to iterate the design to meet the redesign and assessment process required to achieve the results required of the customer's space transportation system. In fact the impact to the schedule to being able to deliver the system that meets the customer's needs, goals, and objectives may cause the customer to compromise his desired operational goal and objectives resulting in considerable

  7. Space Transportation System Availability Requirements and Its Influencing Attributes Relationships

    Rhodes, Russell E.; Adams, Timothy C.; McCleskey, Carey M.

    2008-01-01

    It is important that engineering and management accept the need for an availability requirement that is derived with its influencing attributes. It is the intent of this paper to provide the visibility of relationships of these major attribute drivers (variables) to each other and the resultant system inherent availability. Also important to provide bounds of the variables providing engineering the insight required to control the system's engineering solution, e.g., these influencing attributes become design requirements also. These variables will drive the need to provide integration of similar discipline functions or technology selection to allow control of the total parts count. The relationship of selecting a reliability requirement will place a constraint on parts count to achieve a given availability requirement or if allowed to increase the parts count will drive the system reliability requirement higher. They also provide the understanding for the relationship of mean repair time (or mean down time) to maintainability, e.g., accessibility for repair, and both the mean time between failure, e.g., reliability of hardware and availability. The concerns and importance of achieving a strong availability requirement is driven by the need for affordability, the choice of using the two launch solution for the single space application, or the need to control the spare parts count needed to support the long stay in either orbit or on the surface of the moon. Understanding the requirements before starting the architectural design concept will avoid considerable time and money required to iterate the design to meet the redesign and assessment process required to achieve the results required of the customer's space transportation system. In fact the impact to the schedule to being able to deliver the system that meets the customer's needs, goals, and objectives may cause the customer to compromise his desired operational goal and objectives resulting in considerable

  8. ABCs in College English Education

    李利文

    2011-01-01

    As is known to us all, human beings are always brought under control by such many things as the hormone in our body, the mood produced in our brain and the external world, which leads us to a topic of how to create an atmosphere for the undergraduates to fit well in with the college English education. In our teaching management, the ABCs about college English education works as an assistant. Here, ABCs about college English education refer to some fundamental skills with regard to the undergraduates English education. The ABCs, which always encourage students' involvement, would work effectively if teachers understand them well.

  9. Genome-wide comparative analysis of ABC systems in the Bdellovibrio-and-like organisms.

    Li, Nan; Chen, Huan; Williams, Henry N

    2015-05-10

    Bdellovibrio-and-like organisms (BALOs) are gram-negative, predatory bacteria with wide variations in genome sizes and GC content and ecological habitats. The ATP-binding cassette (ABC) systems have been identified in several prokaryotes, fungi and plants and have a role in transport of materials in and out of cells and in cellular processes. However, knowledge of the ABC systems of BALOs remains obscure. A total of 269 putative ABC proteins were identified in BALOs. The genes encoding these ABC systems occupy nearly 1.3% of the gene content in freshwater Bdellovibrio strains and about 0.7% in their saltwater counterparts. The proteins found belong to 25 ABC system families based on their structural characteristics and functions. Among these, 16 families function as importers, 6 as exporters and 3 are involved in various cellular processes. Eight of these 25 ABC system families were deduced to be the core set of ABC systems conserved in all BALOs. All Bacteriovorax strains have 28 or less ABC systems. On the contrary, the freshwater Bdellovibrio strains have more ABC systems, typically around 51. In the genome of Bdellovibrio exovorus JSS (CP003537.1), 53 putative ABC systems were detected, representing the highest number among all the BALO genomes examined in this study. Unexpected high numbers of ABC systems involved in cellular processes were found in all BALOs. Phylogenetic analysis suggests that the majority of ABC proteins can be assigned into many separate families with high bootstrap supports (>50%). In this study, a general framework of sequence-structure-function connections for the ABC systems in BALOs was revealed providing novel insights for future investigations. PMID:25707746

  10. Politseiniku lustlik ABC / Pekka Erelt

    Erelt, Pekka, 1965-

    1999-01-01

    Politsei on välja andnud 'Politseiniku ABC', mis antakse igale politseinikule. Karikaturist Heiki Ernits on peaaegu igale taskuraamatu leheküljele joonistanud pildikese mundrimeestest kentsakates situatsioonides.

  11. The ABC of Ribosome-Related Antibiotic Resistance

    Wilson, Daniel N.

    2016-01-01

    ABSTRACT The increase in multidrug-resistant pathogenic bacteria is limiting the utility of our current arsenal of antimicrobial agents. Mechanistically understanding how bacteria obtain antibiotic resistance is a critical first step to the development of improved inhibitors. One common mechanism for bacteria to obtain antibiotic resistance is by employing ATP-binding cassette (ABC) transporters to actively pump the drug from the cell. The ABC-F family includes proteins conferring resistance to a variety of clinically important ribosome-targeting antibiotics; however, controversy remains as to whether resistance is conferred via efflux like other ABC transporters or whether another mechanism, such as ribosome protection, is at play. A recent study by Sharkey and coworkers (L. K. Sharkey, T. A. Edwards, and A. J. O’Neill, mBio 7:e01975-15, 2016, http://dx.doi.org/10.1128/mBio.01975-15) provides strong evidence that ABC-F proteins conferring antibiotic resistance utilize ribosome protection mechanisms, namely, by interacting with the ribosome and displacing the drug from its binding site, thus revealing a novel role for ABC-F proteins in antibiotic resistance. PMID:27143393

  12. Analysis of near-term spent fuel transportation hardware requirements and transportation costs

    A computer model was developed to quantify the transportation hardware requirements and transportation costs associated with shipping spent fuel in the commercial nucler fuel cycle in the near future. Results from this study indicate that alternative spent fuel shipping systems (consolidated or disassembled fuel elements and new casks designed for older fuel) will significantly reduce the transportation hardware requirements and costs for shipping spent fuel in the commercial nuclear fuel cycle, if there is no significant change in their operating/handling characteristics. It was also found that a more modest cost reduction results from increasing the fraction of spent fuel shipped by truck from 25% to 50%. Larger transportation cost reductions could be realized with further increases in the truck shipping fraction. Using the given set of assumptions, it was found that the existing spent fuel cask fleet size is generally adequate to perform the needed transportation services until a fuel reprocessing plant (FRP) begins to receive fuel (assumed in 1987). Once the FRP opens, up to 7 additional truck systems and 16 additional rail systems are required at the reference truck shipping fraction of 25%. For the 50% truck shipping fraction, 17 additional truck systems and 9 additional rail systems are required. If consolidated fuel only is shipped (25% by truck), 5 additional rail casks are required and the current truck cask fleet is more than adequate until at least 1995. Changes in assumptions could affect the results. Transportation costs for a federal interim storage program could total about $25M if the FRP begins receiving fuel in 1987 or about $95M if the FRP is delayed until 1989. This is due to an increased utilization of federal interim storage facility from 350 MTU for the reference scenario to about 750 MTU if reprocessing is delayed by two years

  13. Space Transportation System Availability Requirements and Its Influencing Attributes Relationships

    Rhodes, Russel E.; Adams, TImothy C.

    2008-01-01

    It is essential that management and engineering understand the need for an availability requirement for the customer's space transportation system as it enables the meeting of his needs, goal, and objectives. There are three types of availability, e.g., operational availability, achieved availability, or inherent availability. The basic definition of availability is equal to the mean uptime divided by the sum of the mean uptime plus the mean downtime. The major difference is the inclusiveness of the functions within the mean downtime and the mean uptime. This paper will address tIe inherent availability which only addresses the mean downtime as that mean time to repair or the time to determine the failed article, remove it, install a replacement article and verify the functionality of the repaired system. The definitions of operational availability include the replacement hardware supply or maintenance delays and other non-design factors in the mean downtime. Also with inherent availability the mean uptime will only consider the mean time between failures (other availability definitions consider this as mean time between maintenance - preventive and corrective maintenance) that requires the repair of the system to be functional. It is also essential that management and engineering understand all influencing attributes relationships to each other and to the resultant inherent availability requirement. This visibility will provide the decision makers with the understanding necessary to place constraints on the design definition for the major drivers that will determine the inherent availability, safety, reliability, maintainability, and the life cycle cost of the fielded system provided the customer. This inherent availability requirement may be driven by the need to use a multiple launch approach to placing humans on the moon or the desire to control the number of spare parts required to support long stays in either orbit or on the surface of the moon or mars. It is

  14. Differences of Technical Requirements Between Transportation and Storage Metal Casks

    The worldwide demand of storage facilities for spent fuels discharged from nuclear power stations is increasing to maintain sustainable operation of the nuclear power stations. The spent fuels are stored at first in the fuel pools (wet storage). When the spent fuels exceed the pool storage capacity, the fuels are transferred to the other storage facility located at reactor or away from reactor, which often adopts a dry storage technology. To use metal casks is one of the options for the dry storage facilities, and some storage facilities have already utilized large metal casks, whose original design concept were developed to transport the spent fuels from nuclear power stations to a reprocessing plant by trains, trucks or by sea-going vessels. It is widely understood that the technology of transportation casks developed up to now is able to apply to the storage casks without any significant design changes. Technical requirements on the design are discussed between the storage cask and the transportation cask to confirm of the understanding based on the assumption that the large metal cask is used for transportation and storage respectively. (author)

  15. Purification, crystallization and preliminary X-ray analysis of the galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) of the ABC transporter from Bifidobacterium longum JCM1217

    Preliminary X-ray analysis of the galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) of the ABC transporter from B. longum is described. A recombinant galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) from Bifidobacterium longum JCM1217 has been prepared and crystallized by the hanging-drop vapour-diffusion method using 10 mg ml−1 purified enzyme, 0.01 M zinc sulfate, 0.1 M MES buffer pH 5.9–6.4 and 20–22%(v/v) PEG MME 550 in the presence of 5 mM disaccharide ligands. Suitable crystals grew after 10 d incubation at 293 K. The crystals belong to space group C2221, with unit-cell parameters a = 106.3, b = 143.6, c = 114.6 Å for the lacto-N-biose I complex and a = 106.4, b = 143.4, c = 115.5 Å for the galacto-N-biose complex, and diffracted to 1.85 and 1.99 Å resolution, respectively

  16. The multidrug ABC transporter BmrC/BmrD of Bacillus subtilis is regulated via a ribosome-mediated transcriptional attenuation mechanism

    Reilman, Ewoud; Mars, Ruben A. T.; van Dijl, Jan Maarten; Denham, Emma L.

    2014-01-01

    Expression of particular drug transporters in response to antibiotic pressure is a critical element in the development of bacterial multidrug resistance, and represents a serious concern for human health. To obtain a better understanding of underlying regulatory mechanisms, we have dissected the tra

  17. Prognostic impact of high ABC transporter activity in 111 adult acute myeloid leukemia patients with normal cytogenetics when compared to FLT3, NPM1, CEBPA and BAALC.

    Hirsch, Pierre; Tang, Ruoping; Marzac, Christophe; Perrot, Jean-Yves; Fava, Fanny; Bernard, Chantal; Jeziorowska, Dorota; Marie, Jean Pierre; Legrand, Ollivier

    2012-02-01

    ATP-binding cassette transporter (and specially P-glycoprotein) activity is a well known prognostic factor in acute myeloid leukemia, but when compared to other molecular markers its prognostic value has not been well studied. Here we study relationships between this activity, fms-like tyro-sine kinase 3(FLT3/ITD), nucleophosmin(NPM1), CAAT-enhancer binding protein alpha(CEBPα), and brain and acute leukemia cytoplasmic protein (BAALC), in 111 patients with normal cytogenetics who underwent the same treatment, and evaluate its prognostic impact. Independent factors for survival were age (P=0.0126), ATP-binding cassette transporter activity (P=0.018) and duplications in the fms-like tyrosine kinase 3 (P=0.0273). In the 66 patients without fms-like tyrosine kinase 3 duplication and without nucleophosmin mutation, independent prognostic factors for complete remission achievement and survival were age and ATP-binding cassette transporter activity. In conclusion, ATP-binding cassette transporter activity remains an independent prognostic factor, and could assist treatment decisions in patients with no nucleophosmin mutation and no fms-like tyrosine kinase 3 duplication. PMID:22058196

  18. Physical and transportation requirements for a FLIP fueled TRIGA

    Several major changes to the OSTR Physical Security Plan were required by the NRC prior to the August 1976 receipt and installation of a new core consisting entirely of FLIP fuel. The general nature of these changes will be reviewed along with several decisions we faced during their implementation. At the previous TRIGA Owners' Conference in Salt Lake City, Utah, we reported on Oregon's regulatory program for research reactor emergency response planning and physical security. The latter program was of particular interest to us in light of the projected FLIP fuel shipments. The impact of the State's program for physical security of FLIP fuel during transportation will be presented. (author)

  19. Prognostic impact of high ABC transporter activity in 111 adult acute myeloid leukemia patients with normal cytogenetics when compared to FLT3, NPM1, CEBPA and BAALC

    Hirsch, Pierre; Tang, Ruoping; Marzac, Christophe; Perrot, Jean-Yves; FAVA, FANNY; Bernard, Chantal; Jeziorowska, Dorota; MARIE, JEAN PIERRE; Legrand, Ollivier

    2012-01-01

    ATP-binding cassette transporter (and specially P-glycoprotein) activity is a well known prognostic factor in acute myeloid leukemia, but when compared to other molecular markers its prognostic value has not been well studied. Here we study relationships between this activity, fms-like tyro-sine kinase 3(FLT3/ITD), nucleophosmin(NPM1), CAAT-enhancer binding protein alpha(CEBPα), and brain and acute leukemia cytoplasmic protein (BAALC), in 111 patients with normal cytogenetics who underwent th...

  20. Trehalose-recycling ABC transporter LpqY-SugA-SugB-SugC is essential for virulence of Mycobacterium tuberculosis

    Kalscheuer, Rainer; Weinrick, Brian; Veeraraghavan, Usha; Besra, Gurdyal S; Jacobs, William R.

    2010-01-01

    Mycobacterium tuberculosis (Mtb) is an exclusively human pathogen that proliferates within phagosomes of host phagocytes. Host lipids are believed to provide the major carbon and energy sources for Mtb, with only limited availability of carbohydrates. There is an apparent paradox because five putative carbohydrate uptake permeases are present in Mtb, but there are essentially no host carbohydrates inside phagosomes. Nevertheless, carbohydrate transporters have been implicated in Mtb pathogene...

  1. The Intracellular Pathogen Rhodococcus equi Produces a Catecholate Siderophore Required for Saprophytic Growth▿

    Miranda-CasoLuengo, Raúl; Prescott, John F.; Vázquez-Boland, José A.; Meijer, Wim G.

    2007-01-01

    Little is known about the iron acquisition systems of the soilborne facultative intracellular pathogen Rhodococcus equi. We previously reported that expression of iupABC, encoding a putative siderophore ABC transporter system, is iron regulated and required for growth at low iron concentrations. Here we show that disruption of iupA leads to the concomitant accumulation of catecholates and a chromophore with absorption maxima at 341 and 528 nm during growth under iron-replete conditions. In co...

  2. 稻瘟病菌ABC转运蛋白基因中SSR的分布及其功能预测%SSR distribution and its potential effects on function of ABC transporter genes in rice blast fungus Magnaporthe grisea

    刘林; 李成云; 杨静; 苏源; 吴晗; 徐玲; 胡燕玲

    2011-01-01

    ABC transporters play an important role in pathogenicity of fungi. In this study, 47 ABC transporter genes were retrieved from the genome of Magnaporthe grisea. The component and distribution of simple sequence repeats (SSR) within the extron, intron, 5'-UTR and 3'-UTR regions of these genes were analyzed respectively. Furthermore, effects of SSR expansion and contraction on protein structure in the gene coding regions was predicted. The results showed that component and distribution of the SSRs in these regions were different and the SSRs also distributed unevenly in regulation and function regions of these genes. Compared with other regions, the tri-nucleotide and hexa-nucleotide appeared more frequently in coding region. In addition, the G + C component was more abundant and the frequency of hydrophilic amino acids was relatively higher than that of hydrophobic amino acids in the SSRs. Depending on the variation among field isolates of M. Grisea, the expansion or contraction of amino acids encoding SSR sequence were potentially affected on protein secondary structure and gene function. These data implicated that the mutation of SSR might play an important role in variation of pathogenicity-related genes.%ABC转运蛋白在真菌的致病性方面起着重要的作用.本研究查找和分析了稻瘟病菌基因组中47个ABC转运蛋白基因的外显子区、内含子区、5′-UTR和3′-UTR区中的SSR,并对编码区中SSR的扩张或收缩对蛋白结构的影响进行了预测.结果表明,SSR在这些基因的调控区和编码区分布不均一,且在基因的外显子区、内含子区、5′-UTR和3′-UTR区中SSR的组成和分布均不相同;基因的编码区中三碱基和六碱基SSR相对较多,SSR的基序大都表现为GC含量较高,编码的亲水性氨基酸出现频率远远高于疏水性氨基酸.根据稻瘟病菌自然群体中SSR的变化幅度,预测了SSR的扩张或收缩对蛋白二级结构的影响,发现SSR的扩张或收

  3. Development of Fourth Generation ABC Inhibitors from Natural Products: A Novel Approach to Overcome Cancer Multidrug Resistance.

    Karthikeyan, Subburayan; Hoti, Sugeerappa Laxmanappa

    2015-01-01

    Multidrug resistance (MDR) in cancer caused due to overexpression of ABC drug transporters is a major problem in modern chemotherapy. Molecular investigations on MDR have revealed that the resistance is due to various transport proteins of the ABC superfamily which include Phosphoglycoprotein (P-gp/MDR1/ ABCB1), multidrug resistance-associated protein-1 (MRP1), and the breast cancer resistance protein (BCRP). They have been characterized functionally and are considered as major players in the development of MDR in cancer cells. These ATP-dependent transporter proteins cause MDR either by decreased uptake of the drug or increased efflux of the drug from the target organelles. Several MDR-reversing agents are being developed and are in various stages of clinical trials. The first three generations of ABC modulators such as quinine, verapamil, cyclosporine-A, tariquitor, PSC 833, LY335979, and GF120918 required to be administered in high doses to reverse MDR and were associated with adverse effects. Additionally, these modulators non-selectively inhibit ABC and adversely accumulate chemotherapeutic drugs in brain and kidney. Currently, research has stepped up towards reversing MDR by using natural products which exhibitted potential as chemosensitizers. Globally, there is a rich biodiversity of natural products which can be sourced for developing drugs. These products may provide more lead compounds with superior activity, foremost to the development of more effective therapies for MDR cancer cells. Here, we briefly review the status of natural products for reversing MDR modulators, and discuss the long term goal of MDR strategies in current clinical settings. PMID:25584696

  4. The ABCs of Student Engagement

    Parsons, Seth A.; Nuland, Leila Richey; Parsons, Allison Ward

    2014-01-01

    Student engagement is an important consideration for teachers and administrators because it is explicitly associated with achievement. What the authors call the ABC's of engagement they outline as: Affective engagement, Behavioral engagement, and Cognitive engagement. They also present "Three Things Every Teacher Needs to Know about…

  5. 75 FR 11991 - ABC & D Recycling, Inc.-Lease and Operation Exemption-a Line of Railroad in Ware, MA

    2010-03-12

    ... Surface Transportation Board ABC & D Recycling, Inc.--Lease and Operation Exemption--a Line of Railroad in... under 49 CFR 1150.31 to lease from O'Riley Family Trust (O'Riley), and to operate 773 feet of rail line... lease and operate the railroad trackage owned by O'Riley. ABC & D states that it has and intends...

  6. The use of active breathing control (ABC) to reduce margin for breathing motion

    . The variation of the organ positions and volumes for the different scans were quantified and compared. Results: The ABC procedure was well tolerated in the 12 patients. When ABC was applied near the end of normal expiration, the minimal duration of active breath-hold was 15 s for 1 patient with lung cancer, and 20 s or more for all other patients. The duration was greater than 40 s for 2 patients with Hodgkin's disease when ABC was applied during deep inspiration. Scan artifacts associated with normal breathing motion were not observed in the ABC scans. The analysis of the small set of intrafraction scan data indicated that with ABC, the liver volumes were reproducible at about 1%, and lung volumes to within 6%. The excursions of a 'center of target' parameter for the livers were less than 1 mm at the same respiratory phase, but were larger than 4 mm at the extremes of the breathing cycle. The inter-fraction scan study indicated that daily setup variation contributed to the uncertainty in assessing the reproducibility of organ immobilization with ABC between treatment fractions. Conclusion: The results were encouraging; ABC provides a simple means to minimize breathing motion. When applied for CT scanning and treatment, the ABC procedure requires no more than standard operation of the CT scanner or the medical accelerator. The ABC scans are void of motion artifacts commonly seen on fast spiral CT scans. When acquired at different points in the breathing cycle, these ABC scans show organ motion in three-dimension (3D) that can be used to enhance treatment planning. Reproducibility of organ immobilization with ABC throughout the course of treatment must be quantified before the procedure can be applied to reduce margin for conformal treatment

  7. Know Your Diabetes ABCs

    Full Text Available ... Boykin has diabetes. She has suffered from heart disease and required bypass surgery because she was unaware of the link between diabetes and heart disease. Judith Fradkin, M.D.: What most people don' ...

  8. Know Your Diabetes ABCs

    Full Text Available ... Sharon Boykin has diabetes. She has suffered from heart disease and required bypass surgery because she was unaware of the link between diabetes and heart disease. Judith Fradkin, M.D.: What most people don' ...

  9. Know Your Diabetes ABCs

    Full Text Available ... do to correct it all. Announcer: Sharon Boykin has diabetes. She has suffered from heart disease and required bypass surgery ... a measure of how well the blood sugar has been controlled over the previous three months. B ...

  10. Know Your Diabetes ABCs

    Full Text Available Sharon Boykin: From the side effects of the diabetes I've had five strokes. I've had ... to correct it all. Announcer: Sharon Boykin has diabetes. She has suffered from heart disease and required ...

  11. Non-equivalent roles of two periplasmic subunits in the function and assembly of triclosan pump TriABC from Pseudomonas aeruginosa.

    Weeks, Jon W; Nickels, Logan M; Ntreh, Abigail T; Zgurskaya, Helen I

    2015-10-01

    In Gram-negative bacteria, multidrug efflux transporters function in complexes with periplasmic membrane fusion proteins (MFPs) that enable antibiotic efflux across the outer membrane. In this study, we analyzed the function, composition and assembly of the triclosan efflux transporter TriABC-OpmH from Pseudomonas aeruginosa. We report that this transporter possesses a surprising substrate specificity that encompasses not only triclosan but the detergent SDS, which are often used together in antibacterial soaps. These two compounds interact antagonistically in a TriABC-dependent manner and negate antibacterial properties of each other. Unlike other efflux pumps that rely on a single MFP for their activities, two different MFPs, TriA and TriB, are required for triclosan/SDS resistance mediated by TriABC-OpmH. We found that analogous mutations in the α-helical hairpin and membrane proximal domains of TriA and TriB differentially affect triclosan efflux and assembly of the complex. Furthermore, our results show that TriA and TriB function as a dimer, in which TriA is primarily responsible for stabilizing interactions with the outer membrane channel, whereas TriB is important for the stimulation of the transporter. We conclude that MFPs are engaged into complexes as asymmetric dimers, in which each protomer plays a specific role. PMID:26193906

  12. 19 CFR 122.117 - Requirements for transit air cargo transport.

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Requirements for transit air cargo transport. 122... Requirements for transit air cargo transport. (a) Transportation—(1) Port to port. Transit air cargo may be... cargo, a receipt shall be given. The receipt shall be made by the airline responsible for transport...

  13. Applying ABC analysis to the Navy's inventory management system

    May, Benjamin

    2014-01-01

    Approved for public release; distribution is unlimited ABC Analysis is an inventory categorization technique used to classify and prioritize inventory items in an effort to better allocate business resources. A items are defined as the inventory items considered extremely important to the business, requiring strict oversight and control. B items are important to the business, but don’t require the tight controls and oversight required of the A items. C items are marginally important to the...

  14. Know Your Diabetes ABCs

    Full Text Available ... I've had eight stents put into my heart. I've had open heart surgery, partial amputation of both feet. I have ... Sharon Boykin has diabetes. She has suffered from heart disease and required bypass surgery because she was ...

  15. Know Your Diabetes ABCs

    Full Text Available ... amputation of both feet. I have cataracts and glaucoma now and high blood pressure and I have high cholesterol. That's before I found out what I needed to do to correct it all. Announcer: Sharon Boykin has diabetes. She has suffered from heart disease and required ...

  16. Know Your Diabetes ABCs

    Full Text Available ... into my heart. I've had open heart surgery, partial amputation of both feet. I have cataracts and glaucoma now and high blood pressure and I have high cholesterol. That's before I found out what I needed to do to ... and required bypass surgery because she was unaware of the link between ...

  17. Key requirements of packet transport network based on MPLS-TP

    Huang, Feng; Yi, Xiaobo; Zhang, Hanzheng; Gong, Ping

    2009-11-01

    Requirement of packet transport network based on MPLS-TP are analyzed including in transport plane, OAM, survivability, QoS, control plane and management plane. MPLS-TP standard status is also introduced.

  18. The use of active breathing control (ABC) to minimize breathing motion during radiation therapy

    or during deep inspiration; and one ABC scan near the end of normal expiration. For two patients, the ABC scans were repeated 1 week later. Contours of the lungs, liver, and kidneys were outlined when applicable. The positions and volumes of the organs in the different scans were quantified and compared. Results. The ABC procedure was tolerated well by all patients. When ABC was applied near the end of normal expiration, the minimal duration of active breath-hold was 15 seconds for a lung patient, and 20 seconds or more for all other patients. Two patients maintained 50 seconds of breath-hold when ABC was applied during deep inspiration. The usual scan artifacts associated with breathing motion were not observed in the ABC scans. The volumes of soft tissue organs were similar for the ABC scans. The positions of the liver and kidney varied by about 1.5 cm and 0.5 cm respectively between the end of normal inspiration and expiration. Organ positions were highly reproducible for identical points in the breathing cycle. Lung volumes varied by less than 5% in ABC scans that were acquired during the same CT session and when they were separated by 1 week. Figure 1 shows an example of the almost identical diaphragm positions of a normal subject in 2 ABC 'scout-view' profiles taken 5 minutes apart. Conclusions. The results are promising. ABC provides a simple means to reproducibly minimize breathing motion. When applied for treatment, the ABC procedure requires no more than normal operation of the treatment machine. The minimal variation in organ position with repeated application of ABC indicates that it is possible to complete, piece-meal, a treatment that might require extended beam-on time. Research is being conducted to examine the variation of breathing motion during the course of treatment and to incorporate the information in treatment planning

  19. Sustaining an Effective ABC-ABM System

    Gary COKINS

    2011-02-01

    Full Text Available The purpose of this paper is to describe the Activity- Based Costing (ABC and Activity-Based Management (ABM system and techniques to sustain them as a permanent and repeatable production reporting system, not just for one-off analysis. A comparison is made between ABC/ABM modeling software that extracts source data and business systems that include ABC/ABM modeling features. There are presented the stages of updating, running and rerunning the ABC/ABM system. The resulting information calculated and provided by the ABC/ABM system are analyzed and interpreted in terms of a multidimensional data analysis. The article ends with the authors' conclusions about the benefits of continued operation of sustaining the ABC/ABM system.

  20. Transport of radioactive materials and equipment. Requirements. (Provisional)

    This standard is aimed at establishing the procedures that must be followed when transporting radioactive materials and equipment in Venezuelan Territory. The ''Consejo Nacional para el Desarrollo de la Industria Nuclear'' is responsible for their fulfillment and control

  1. Application of activity-based costing (ABC) for a Peruvian NGO healthcare provider.

    Waters, H; Abdallah, H; Santillán, D

    2001-01-01

    This article describes the application of activity-based costing (ABC) to calculate the unit costs of the services for a health care provider in Peru. While traditional costing allocates overhead and indirect costs in proportion to production volume or to direct costs, ABC assigns costs through activities within an organization. ABC uses personnel interviews to determine principal activities and the distribution of individual's time among these activities. Indirect costs are linked to services through time allocation and other tracing methods, and the result is a more accurate estimate of unit costs. The study concludes that applying ABC in a developing country setting is feasible, yielding results that are directly applicable to pricing and management. ABC determines costs for individual clinics, departments and services according to the activities that originate these costs, showing where an organization spends its money. With this information, it is possible to identify services that are generating extra revenue and those operating at a loss, and to calculate cross subsidies across services. ABC also highlights areas in the health care process where efficiency improvements are possible. Conclusions about the ultimate impact of the methodology are not drawn here, since the study was not repeated and changes in utilization patterns and the addition of new clinics affected applicability of the results. A potential constraint to implementing ABC is the availability and organization of cost information. Applying ABC efficiently requires information to be readily available, by cost category and department, since the greatest benefits of ABC come from frequent, systematic application of the methodology in order to monitor efficiency and provide feedback for management. The article concludes with a discussion of the potential applications of ABC in the health sector in developing countries. PMID:11326572

  2. No facilitator required for membrane transport of hydrogen sulfide

    Mathai, John C.; Missner, Andreas; Kügler, Philipp; Saparov, Sapar M.; Zeidel, Mark L.; Lee, John K.; Pohl, Peter

    2009-01-01

    Hydrogen sulfide (H2S) has emerged as a new and important member in the group of gaseous signaling molecules. However, the molecular transport mechanism has not yet been identified. Because of structural similarities with H2O, it was hypothesized that aquaporins may facilitate H2S transport across cell membranes. We tested this hypothesis by reconstituting the archeal aquaporin AfAQP from sulfide reducing bacteria Archaeoglobus fulgidus into planar membranes and by monitoring the resulting fa...

  3. ABC model and floral evolution

    LI Guisheng; MENG Zheng; KONG Hongzhi; CHEN Zhiduan; LU Anming

    2003-01-01

    The paper introduces the classical ABC model of floral development and thereafter ABCD, ABCDE and quartet models, and presents achievements in the studies on floral evolution such as the improved understanding on the relationship of reproductive organs between gnetophytes and angiosperms, new results in perianth evolution and identified homology of floral organs between dicots and monocots. The evo-devo studies on plant taxa at different evolutionary levels are useful to better understanding the homology of floral organs, and to clarifying the mysteries of the origin and subsequent diversification of flowers.

  4. Learning the ABCs: Activity based costing in waste operations

    The United States Department of Energy (DOE) is facing a challenging new national role based on current world events, changing public perception and awareness, and a legacy of wastes generated in the past. Clearly, the DOE must put mechanisms in place to comply with environmental rules, regulations, and good management practices so that public health risk is minimized while programmatic costs are controlled. DOE has begun this process and has developed a Five-Year Plan to describe the activities necessary to comply with both cleanup, or environmental restoration, and waste management of existing waste streams. The focus of this paper is how to best manage the treatment, storage, disposal, and transportation of waste throughout the DOE weapons complex by using Activity Based Costing (ABC) to both plan and control expenditures in DOE Waste Management (WM). The basics of ABC, along with an example, will be detailed. (author)

  5. The influence of contract form choice of the transport services ensuring on performance contracting entity requirement

    Miloš POLIAK; Anna KRIŽANOVÁ; Štefánia SEMANOVÁ; Ľubica ŠTEFÁNIKOVÁ

    2014-01-01

    The paper deals with the issue of the influence of contract form choice of the transport services ensuring on performance contracting entity requirement. In the first part, it describes the current state of transport services financing in the Slovak Republic and risk analysis, which is related to conclusion of the service contract in the transport services ensuring. The rest of the paper defines the impact of services contracts of ensuring the transport services on performance contracting ent...

  6. 30 CFR 75.1106-2 - Transportation of liquefied and nonliquefied compressed gas cylinders; requirements.

    2010-07-01

    ... compressed gas cylinders; requirements. 75.1106-2 Section 75.1106-2 Mineral Resources MINE SAFETY AND HEALTH... Fire Protection § 75.1106-2 Transportation of liquefied and nonliquefied compressed gas cylinders; requirements. (a) Liquefied and nonliquefied compressed gas cylinders transported into or through...

  7. Dopamine transporter inhibition is required for cocaine-induced stereotypy

    Tilley, Michael R.; Gu, Howard H.

    2008-01-01

    The primary mechanism by which cocaine induces stereotypy has been difficult to discern because cocaine has three high affinity targets, the reuptake transporters for dopamine (DAT), norepinephrine, and serotonin. To dissect out the role of DAT in cocaine effects, we generated a knock-in mouse line with a cocaine insensitive DAT (DAT-CI mice). DAT-CI mice provide a powerful tool that can directly test whether DAT inhibition is important for cocaine-induced stereotypy. We found that acute coca...

  8. Phosphatidylserine flipping enhances membrane curvature and negative charge required for vesicular transport

    Xu, Peng; Baldridge, Ryan D.; Chi, Richard J.; Burd, Christopher G.; Graham, Todd R.

    2013-01-01

    Vesicle-mediated protein transport between organelles of the secretory and endocytic pathways is strongly influenced by the composition and organization of membrane lipids. In budding yeast, protein transport between the trans-Golgi network (TGN) and early endosome (EE) requires Drs2, a phospholipid translocase in the type IV P-type ATPase family. However, downstream effectors of Drs2 and specific phospholipid substrate requirements for protein transport in this pathway are unknown. Here, we ...

  9. Global energy transports and the influence of clouds on transport requirements - A satellite analysis

    Sohn, Byung-Ju; Smith, Eric A.

    1992-01-01

    This report investigates the impact of differential net radiative heating on 2D energy transports within the atmosphere ocean system and the role of clouds on this process. The 2D mean energy transports, in answer to zonal and meridional gradients in the net radiation field, show an east-west coupled dipole structure in which the Pacific acts as the major energy source and North Africa as the major energy sink. It is demonstrated that the dipole is embedded in the secondary energy transports arising mainly from the differential heating between land and oceans in the tropics in which the tropical east-west (zonal) transports are up to 30 percent of the tropical north-south (meridional) transports.

  10. Inventory management for the health sector: ABC analysis approach

    Nabais, Joana Isabel Baptista

    2010-01-01

    A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics This project aims to analyse a hospital’s inventory management and make suggestions to improve its practices, with special attention on ABC analysis as an optimization tool for the inventory management, control and storage. Other cost reductions approaches are studied in order to contribute for the accurate management of clinical consumption...

  11. Greater-than-Class C low-level radioactive waste transportation regulations and requirements study

    The purpose of this report is to identify the regulations and requirements for transporting greater-than-Class C (GTCC) low-level radioactive waste (LLW) and to identify planning activities that need to be accomplished in preparation for transporting GTCC LLW. The regulations and requirements for transporting hazardous materials, of which GTCC LLW is included, are complex and include several Federal agencies, state and local governments, and Indian tribes. This report is divided into five sections and three appendices. Section 1 introduces the report. Section 2 identifies and discusses the transportation regulations and requirements. The regulations and requirements are divided into Federal, state, local government, and Indian tribes subsections. This report does not identify the regulations or requirements of specific state, local government, and Indian tribes, since the storage, treatment, and disposal facility locations and transportation routes have not been specifically identified. Section 3 identifies the planning needed to ensure that all transportation activities are in compliance with the regulations and requirements. It is divided into (a) transportation packaging; (b) transportation operations; (c) system safety and risk analysis, (d) route selection; (e) emergency preparedness and response; and (f) safeguards and security. This section does not provide actual planning since the details of the Department of Energy (DOE) GTCC LLW Program have not been finalized, e.g., waste characterization and quantity, storage, treatment and disposal facility locations, and acceptance criteria. Sections 4 and 5 provide conclusions and referenced documents, respectively

  12. Functional requirements for the Automated Transportation Management System: TTP number: RL 439002

    Portsmouth, J.H. [Westinghouse Hanford Co., Richland, WA (United States)

    1992-12-31

    This requirements analysis, documents Department of Energy (DOE) transportation management procedures for the purpose of providing a clear and mutual understanding between users and designers of the proposed Automated Transportation Management System (ATMS). It is imperative that one understand precisely how DOE currently performs traffic management tasks; only then can an integrated system be proposed that successfully satisfies the major requirements of transportation managers and other system users. Accordingly, this report describes the current workings of DOE transportation organizations and then proposes a new system which represents a synthesis of procedures (both current and desired) which forms the basis for further systems development activities.

  13. An ABC for decision making

    Garcia, Luiz Henrique Costa, E-mail: luiz_mogi@yahoo.com.br [Associacao de Medicina Intensiva Brasileira (AMIB), Sao Paulo, SP (Brazil); Irmandade da Santa Casa de Misericordia de Sao Paulo, SP (Brazil); Ferreira, Bruna Cortez [Hospital de Base de Sao Jose do Rio Preto, SP (Brazil)

    2015-03-15

    The present study was aimed at proposing a systematic evaluation of cranial computed tomography, identifying the main aspects to be analyzed in order to facilitate the decision making process regarding diagnosis and management in emergency settings. The present descriptive study comprised a literature review at the following databases: Access Medicine and Access Emergency Medicine (McGraw-Hill Education); British Medical Journal Evidence Center; UptoDate; Bireme; PubMed; Lilacs; SciELO; ProQuest; Micromedex (Thomson Reuters); Embase. Once the literature review was completed, the authors identified the main diseases with tomographic repercussions and proposed the present system to evaluate cranial computed tomography images. An easy-to-memorize ABC system will facilitate the decision making in emergency settings, as it covers the main diseases encountered by intensivists and emergency physicians, and provides a sequential guidance about anatomical structures to be investigated as well as their respective alterations. (author)

  14. An ABC for decision making.

    Garcia, Luiz Henrique Costa; Ferreira, Bruna Cortez

    2015-01-01

    The present study was aimed at proposing a systematic evaluation of cranial computed tomography, identifying the main aspects to be analyzed in order to facilitate the decision making process regarding diagnosis and management in emergency settings. The present descriptive study comprised a literature review at the following databases: Access Medicine and Access Emergency Medicine (McGraw- Hill Education); British Medical Journal Evidence Center; UptoDate; Bireme; PubMed; Lilacs; SciELO; ProQuest; Micromedex (Thomson Reuters); Embase. Once the literature review was completed, the authors identified the main diseases with tomographic repercussions and proposed the present system to evaluate cranial computed tomography images. An easy-to-memorize ABC system will facilitate the decision making in emergency settings, as it covers the main diseases encountered by intensivists and emergency physicians, and provides a sequential guidance about anatomical structures to be investigated as well as their respective alterations. PMID:25987751

  15. An ABC for decision making

    The present study was aimed at proposing a systematic evaluation of cranial computed tomography, identifying the main aspects to be analyzed in order to facilitate the decision making process regarding diagnosis and management in emergency settings. The present descriptive study comprised a literature review at the following databases: Access Medicine and Access Emergency Medicine (McGraw-Hill Education); British Medical Journal Evidence Center; UptoDate; Bireme; PubMed; Lilacs; SciELO; ProQuest; Micromedex (Thomson Reuters); Embase. Once the literature review was completed, the authors identified the main diseases with tomographic repercussions and proposed the present system to evaluate cranial computed tomography images. An easy-to-memorize ABC system will facilitate the decision making in emergency settings, as it covers the main diseases encountered by intensivists and emergency physicians, and provides a sequential guidance about anatomical structures to be investigated as well as their respective alterations. (author)

  16. An ABC for decision making

    Luiz Henrique Costa Garcia

    2015-04-01

    Full Text Available The present study was aimed at proposing a systematic evaluation of cranial computed tomography, identifying the main aspects to be analyzed in order to facilitate the decision making process regarding diagnosis and management in emergency settings. The present descriptive study comprised a literature review at the following databases: Access Medicine and Access Emergency Medicine (McGraw- Hill Education; British Medical Journal Evidence Center; UptoDate; Bireme; PubMed; Lilacs; SciELO; ProQuest; Micromedex (Thomson Reuters; Embase. Once the literature review was completed, the authors identified the main diseases with tomographic repercussions and proposed the present system to evaluate cranial computed tomography images. An easy-to-memorize ABC system will facilitate the decision making in emergency settings, as it covers the main diseases encountered by intensivists and emergency physicians, and provides a sequential guidance about anatomical structures to be investigated as well as their respective alterations.

  17. Brassboard Astrometric Beam Combiner (ABC) Development for the Space Interferometry Mission (SIM)

    Jeganathan, Muthu; Kuan, Gary; Rud, Mike; Lin, Sean; Sutherland, Kristen; Moore, James; An, Xin

    2008-01-01

    The Astrometric Beam Combiner (ABC) is a critical element of the Space Interferometry Mission (SIM) that performs three key functions: coherently combine starlight from two siderostats; individually detect starlight for angle tracking; and disperse and detect the interferometric fringes. In addition, the ABC contains: a stimulus, cornercubes and shutters for in-orbit calibration; several tip/tilt mirror mechanisms for in-orbit alignment; and internal metrology beam launcher for pathlength monitoring. The detailed design of the brassboard ABC (which has the form, fit and function of the flight unit) is complete, procurement of long-lead items is underway, and assembly and testing is expected to be completed in Spring 2009. In this paper, we present the key requirements for the ABC, details of the completed optical and mechanical design as well as plans for assembly and alignment.

  18. abc: An extensible AspectJ compiler

    Avgustinov, Pavel; Christensen, Aske Simon; Hendren, Laurie;

    2005-01-01

    checking and code generation, as well as data flow and control flow analyses. The AspectBench Compiler (abc) is an implementation of such a workbench. The base version of abc implements the full AspectJ language. Its frontend is built, using the Polyglot framework, as a modular extension of the Java...... language. The use of Polyglot gives flexibility of syntax and type checking. The backend is built using the Soot framework, to give modular code generation and analyses. In this paper, we outline the design of abc, focusing mostly on how the design supports extensibility. We then provide a general overview...

  19. Help guide for the application of regulatory requirements on the transport of radioactive material

    The regulation of transport of radioactive material by road in Spain refers to compliance with the requirements of the European Agreement concerning the International Carriage of Dangerous Goods by Road (ADR) in force. The structure presented by this legislation, which is international, and the inclusion in it of requirements that apply to other dangerous goods makes it difficult to consult the requirements that specifically apply to the transport of radioactive material. Therefore, the Nuclear Safety Council (CSN) has found it necessary to publish a guide that facilitates users to comply with its provisions and, consequently, this transport security.

  20. Performance Optimization of Benchmark Functions Using VTS-ABC Algorithm

    Twinkle Gupta

    2014-10-01

    Full Text Available A new variant based on tournament selection called VTS-ABC algorithm is provided in this paper. Its performance is compared with standard ABC algorithm with different size of data on several Benchmark functions and results show that VTS-ABC provides better quality of solution than original ABC algorithm in every case.

  1. 78 FR 17781 - Transportation Worker Identification Credential (TWIC)-Reader Requirements

    2013-03-22

    ... to work with the Transportation Worker Identification Credential (TWIC) as an access control measure... access control requirements and other measures designed to enhance maritime security. This rulemaking... Physical Access Control System PIN Personal Identification Number QTL Qualified Technology List...

  2. ABC of childhood memories [artists' book

    Wood, Philippa; MacLellan, Tamar

    2009-01-01

    The limited edition artists' book by Philippa Wood and Tamar MacLellan – ABC of Childhood Memories was a collaborative project that explored the 1960s/70s childhood memories of each artist through text based narrative and image.

  3. The abc-conjecture for Algebraic Numbers

    Jerzy BROWKIN

    2006-01-01

    The abc-conjecture for the ring of integers states that, for every ε> 0 and every triple of relatively prime nonzero integers (a, b, c) satisfying a + b = c, we have max(|a|, |b|, |c|) ≤ rad(abc)1+ε with a finite number of exceptions. Here the radical rad(m) is the product of all distinct prime factors of m.In the present paper we propose an abc-conjecture for the field of all algebraic numbers. It is based on the definition of the radical (in Section 1) and of the height (in Section 2) of an algebraic number.From this abc-conjecture we deduce some versions of Fermat's last theorem for the field of all algebraic numbers, and we discuss from this point of view known results on solutions of Fermat's equation in fields of small degrees over Q.

  4. Capital requirements for the transportation of energy materials: 1979 arc estimates

    1980-08-29

    Summaries of transportation investment requirements through 1990 are given for the low, medium and high scenarios. Total investment requirements for the three modes and the three energy commodities can accumulate to a $46.3 to $47.0 billion range depending on the scenario. The high price of oil, following the evidence of the last year, is projected to hold demand for oil below the recent past. Despite the overall decrease in traffic some investment in crude oil and LPG pipelines is necessary to reach new sources of supply. Although natural gas production and consumption is projected to decline through 1990, new investments in carrying capacity also are required due to locational shifts in supply. The Alaska Natural Gas Transportation System is the dominant investment for energy transportation in the next ten years. This year's report focuses attention on waterborne coal transportation to the northeast states in keeping with a return to significant coal consumption projected for this area. A resumption of such shipments will require a completely new fleet. The investment estimates given in this report identify capital required to transport projected energy supplies to market. The requirement is strategic in the sense that other reasonable alternatives do not exist or that a shared load of new growth can be expected. Not analyzed or forecasted are investments in transportation facilities made in response to local conditions. The total investment figures, therefore, represent a minimum necessary capital improvement to respond to changes in interregional supply conditions.

  5. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development.

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L

    2016-01-01

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development. PMID:27381418

  6. Programmatic and technical requirements for the FMDP fresh MOX fuel transport package

    Ludwig, S. B.; Michelhaugh, R. D.; Pope, R. B.; Shappert, L. B.; Singletary, B. H.; Chae, S. M.; Parks, C. V.; Broadhead, B. L.; Schmid, S. P.; Cowart, C. G.

    1997-12-01

    This document is intended to guide the designers of the package to all pertinent regulatory and other design requirements to help ensure the safe and efficient transport of the weapons-grade (WG) fresh MOX fuel under the Fissile Materials Disposition Program. To accomplish the disposition mission using MOX fuel, the unirradiated MOX fuel must be transported from the MOX fabrication facility to one or more commercial reactors. Because the unirradiated fuel contains large quantities of plutonium and is not sufficient radioactive to create a self-protecting barrier to deter the material from theft, DOE intends to use its fleet of safe secure trailers (SSTs) to provide the necessary safeguards and security for the material in transit. In addition to these requirements, transport of radioactive materials must comply with regulations of the Department of Transportation and the Nuclear Regulatory Commission (NRC). In particular, NRC requires that the packages must meet strict performance requirements. The requirements for shipment of MOX fuel (i.e., radioactive fissile materials) specify that the package design is certified by NRC to ensure the materials contained in the packages are not released and remain subcritical after undergoing a series of hypothetical accident condition tests. Packages that pass these tests are certified by NRC as a Type B fissile (BF) package. This document specifies the programmatic and technical design requirements a package must satisfy to transport the fresh MOX fuel assemblies.

  7. Programmatic and technical requirements for the FMDP fresh MOX fuel transport package

    This document is intended to guide the designers of the package to all pertinent regulatory and other design requirements to help ensure the safe and efficient transport of the weapons-grade (WG) fresh MOX fuel under the Fissile Materials Disposition Program. To accomplish the disposition mission using MOX fuel, the unirradiated MOX fuel must be transported from the MOX fabrication facility to one or more commercial reactors. Because the unirradiated fuel contains large quantities of plutonium and is not sufficient radioactive to create a self-protecting barrier to deter the material from theft, DOE intends to use its fleet of safe secure trailers (SSTs) to provide the necessary safeguards and security for the material in transit. In addition to these requirements, transport of radioactive materials must comply with regulations of the Department of Transportation and the Nuclear Regulatory Commission (NRC). In particular, NRC requires that the packages must meet strict performance requirements. The requirements for shipment of MOX fuel (i.e., radioactive fissile materials) specify that the package design is certified by NRC to ensure the materials contained in the packages are not released and remain subcritical after undergoing a series of hypothetical accident condition tests. Packages that pass these tests are certified by NRC as a Type B fissile (BF) package. This document specifies the programmatic and technical design requirements a package must satisfy to transport the fresh MOX fuel assemblies

  8. Regulations for the safe transport of radioactive material. 2009 ed. Safety requirements

    The objective of these Regulations is to establish requirements that must be satisfied to ensure safety and to protect persons, property and the environment from the effects of radiation in the transport of radioactive material. This protection is achieved by requiring: (a) Containment of the radioactive contents; (b) Control of external radiation levels; (c) Prevention of criticality; and (d) Prevention of damage caused by heat. These requirements are satisfied firstly by applying a graded approach to contents limits for packages and conveyances and to performance standards applied to package designs, depending upon the hazard of the radioactive contents. Secondly, they are satisfied by imposing requirements on the design and operation of packages and on the maintenance of packagings, including consideration of the nature of the radioactive contents. Finally, they are satisfied by requiring administrative controls, including, where appropriate, approval by competent authorities. In the transport of radioactive material, the safety of persons and the protection of property and the environment are assured when these Regulations are complied with. Confidence in this regard is achieved through quality assurance and compliance assurance programmes. These Regulations apply to the transport of radioactive material by all modes on land, water, or in the air, including transport which is incidental to the use of the radioactive material. Transport comprises all operations and conditions associated with, and involved in, the movement of radioactive material; these include the design, manufacture, maintenance and repair of packaging, and the preparation, consigning, loading, carriage including in-transit storage, unloading and receipt at the final destination of loads of radioactive material and packages. A graded approach is applied in specifying the performance standards in these Regulations which are characterized in terms of three general severity levels: (a) Routine

  9. National Ignition Facility subsystem design requirements transportation and handling, SSDR 1.1.1.3.2

    This Subsystem Design Requirement document is a development specification that establishes the performance, design, development, and test requirements for the Transportation ampersand Material Handling Systems (WBS 1.1.1.3.2) of the NIF Laser System (WBS 1.3 and 1.4). The NIF is a multi-pass, 192-beam, high-power, neodymium-glass laser that meets requirements set forth in the NIF SDR 002 (Laser System). 5 figs

  10. Requirement of nucleotide exchange factor for Ypt1 GTPase mediated protein transport

    1995-01-01

    Small GTPases of the rab family are involved in the regulation of vesicular transport. It is believed that cycling between the GTP- and GDP-bound forms, and accessory factors regulating this cycling are crucial for rab function. However, an essential role for rab nucleotide exchange factors has not yet been demonstrated. In this report we show the requirement of nucleotide exchange factor activity for Ypt1 GTPase mediated protein transport. The Ypt1 protein, a member of the rab family, plays ...

  11. Competence and Capacity-Building Requirements in Transport and Logistics Market

    Bazaras Darius; Čižiūnienė Kristina; Palšaitis Ramūnas; Kabashkin Igor

    2016-01-01

    The article analyses theoretical aspects of the qualitative assessment of logistics and transport specialists’ potential, as well as provides research results on competence and capacity-building process in preparation of logistics and transport specialists. The results enabled to identify problematic areas of graduate integration in the labour market and foresee the possible requirements for professional development. In some cases, internal motivation is related to problematic points or negat...

  12. The implementation of radiation protection programme requirements in the transport of nuclear fuel cycle materials

    The IAEA requires organisations involved in transport of radioactive material to implement a Radiation Protection Programme (RPP) to control radiation dose exposure to both workers and the public from transport operations. For nuclear fuel cycle materials, radiation protection is dealt with prior to shipment by using a package design to control exposures to workers and the public. Dose assessment and evaluation is a key issue for RPPs. The extent of control measures in the RPP should relate to the magnitude and likelihood of radiation exposure. The World Nuclear Transport Institute (WNTI) therefore made an assessment of the likely doses to workers in the transport chain, for the various modes of transport and for the main nuclear materials. Analysis of the data on dose up-take during the various modes of transport of nuclear fuel cycle materials indicates that it is very unlikely that any group of workers, or any member of the public, will receive annual doses in excess of 1mSv and the transport of nuclear fuel cycle materials should therefore fall into the lowest category, for which no workplace or individual dose monitoring is required. (author)

  13. An ABC analysis for power generation project

    Batool Hasani

    2013-07-01

    Full Text Available One of the primary concerns on performance measurement is to know how much a particular project cost. However, using traditional method on project-based products often leads to inappropriate results. In this paper, we re-examine this issue by comparing the cost of a power station construction project using ABC versus traditional method. The results of survey show that ABC method is capable of providing better estimates for overhead costs compared with traditional method. In other words, ABC method helps reduce some of the unnecessary overhead cost items and increase on some other cost components. This helps increase the relative efficiency of the system by reducing total cost of project.

  14. Establishing Bedding Requirements during Transport and Monitoring Skin Temperature during Cold and Mild Seasons after Transport for Finishing Pigs

    John McGlone

    2014-05-01

    Full Text Available The broad aim of this study was to determine whether bedding level in the transport trailer influenced pig performance and welfare. Specifically, the objective was to define the bedding requirements of pigs during transportation in commercial settings during cold and mild weather. Animals (n = 112,078 pigs on 572 trailers used were raised in commercial finishing sites and transported in trailers to commercial processing plants. Dead on arrival (DOA, non-ambulatory (NA, and total dead and down (D&D data were collected and skin surface temperatures of the pigs were measured by infrared thermography. Data were collected during winter (Experiment 1 and fall/spring (Experiment 2. Total D&D percent showed no interaction between bedding level and outside air temperature in any experiments. Average skin surface temperature during unloading increased with outside air temperature linearly in both experiments (P < 0.01. In conclusion, over-use of bedding may be economically inefficient. Pig skin surface temperature could be a useful measure of pig welfare during or after transport.

  15. Accelerator-based conversion (ABC) of reactor and weapons plutonium

    An accelerator-based conversion (ABC) system is presented that is capable of rapidly burning plutonium in a low-inventory sub-critical system. The system also returns fission power to the grid and transmutes troublesome long-lived fission products to short lived or stable products. Higher actinides are totally fissioned. The system is suited not only to controlled, rapid burning of excess weapons plutonium, but to the long range application of eliminating or drastically reducing the world total inventory of plutonium. Deployment of the system will require the successful resolution of a broad range of technical issues introduced in the paper

  16. Accelerator-based conversion (ABC) of reactor and weapons plutonium

    Jensen, R.J.; Trapp, T.J.; Arthur, E.D.; Bowman, C.D.; Davidson, J.W.; Linford, R.K.

    1993-06-01

    An accelerator-based conversion (ABC) system is presented that is capable of rapidly burning plutonium in a low-inventory sub-critical system. The system also returns fission power to the grid and transmutes troublesome long-lived fission products to short lived or stable products. Higher actinides are totally fissioned. The system is suited not only to controlled, rapid burning of excess weapons plutonium, but to the long range application of eliminating or drastically reducing the world total inventory of plutonium. Deployment of the system will require the successful resolution of a broad range of technical issues introduced in the paper.

  17. Analisis Incremental Kelayakan Penambahan Lini Perakitan Engine Motor PT ABC

    Jonny

    2013-01-01

    Manufacturing motorcycle unit in Plant 1 of PT ABC requires engine supply from Plant 2. This has some burden on the operating cost of Plant 1 and the opportunity loss as well due to delay of motor engine delivery. Based on the problem, team proposes additional engine line in Plant 1 to cut engine supply from Plant 2. Therefore, team analyzes this proposal using incremental analysis to determine whether the proposal is feasible or not by comparing before and after condition with result NPV abo...

  18. The ABC-paradox: is Time Driven ABC relevant for small and Medium sized enterprises (SME)?

    Fladkjær, Henrik Find; Jensen, Erling

    Several articles suggest that Activity Based Costing (ABC) has failed to succeed in practical use. It is even argued that we have an ABC-paradox. Activity Based Costing has won theoretically in nu-merous articles in journals, through books, being included in all major Business Accounting text-books...... manufacturing company we will analyse whether it is possible to create a satisfactory decision making foundation using a TDABC as a less complex cost model in the company. We have analysed our way to the conclusion that compared to ABC, the TDABC model provides the management with a number of pragmatic...

  19. 49 CFR 173.427 - Transport requirements for low specific activity (LSA) Class 7 (radioactive) materials and...

    2010-10-01

    ... this section must be packaged in accordance with 10 CFR part 71. (e) Tables 5 and 6 are as follows... 49 Transportation 2 2010-10-01 2010-10-01 false Transport requirements for low specific activity... SHIPMENTS AND PACKAGINGS Class 7 (Radioactive) Materials § 173.427 Transport requirements for low...

  20. A note on ABC implementation: An evaluation case.

    Nur Naha Abu Mansor

    2004-01-01

    Whilst featuring prominently in management accounting research for at least the last decade, activity-based costing (ABC) may have reached a plateau in term of implementation and success. This paper reports the results of a survey of activity-based costing (ABC) in a telecommunication organization. The study considers ABC information and systems being used in various parts of division of the company. It reports on success measures adapted from the Foster and Swenson (1997) survey of ABC imple...

  1. Approximate maximum likelihood estimation using data-cloning ABC

    Picchini, Umberto; Anderson, Rachele

    2015-01-01

    A maximum likelihood methodology for a general class of models is presented, using an approximate Bayesian computation (ABC) approach. The typical target of ABC methods are models with intractable likelihoods, and we combine an ABC-MCMC sampler with so-called "data cloning" for maximum likelihood estimation. Accuracy of ABC methods relies on the use of a small threshold value for comparing simulations from the model and observed data. The proposed methodology shows how to use large threshold ...

  2. Transition fibre protein FBF1 is required for the ciliary entry of assembled intraflagellar transport complexes

    WEI, QING; Xu, Qingwen; Zhang, Yuxia; Li, Yujie; Zhang, Qing; Hu, Zeng; Harris, Peter C.; Torres, Vicente E.; Ling, Kun; Hu, Jinghua

    2013-01-01

    Sensory organelle cilia play critical roles in mammalian embryonic development and tissue homeostasis. Intraflagellar transport (IFT) machinery is required for the assembly and maintenance of cilia. Yet how this large complex passes through the size-dependent barrier at the ciliary base remains enigmatic. Here we report that FBF1, a highly conserved transition fibre protein, is required for the ciliary import of assembled IFT particles at the cilia base. We cloned dyf-19, the C. elegans homol...

  3. Commercial Space Transportation and Air Traffic Insertion - SESAR Requirements and the European Perspective

    Drescher, Jürgen; Morlang, Frank; Hampe, Jens; Kaltenhäuser, Sven; Jakobi, Jörn; Schmitt, Dirk-Roger

    2016-01-01

    Commercial Space Transportation becomes an international business and requires landing opportunities all over the world. Hence the integration of space vehicles in other airspace than the US NAS is an important topic to be considered. The Single European Sky ATM Research Programme (SESAR) is preparing the implementation of a new ATM system in Europe. The requirements are defined by the concept of the shared Business Trajectory and System Wide Information Management (SWIM).This paper describes...

  4. Engineering Requirements for Offshore CO₂ Transportation and Storage: A Summary Based on International Experiences

    Haszeldine, R. Stuart; Zhou, Di; Zhang, Yunfang

    2014-01-01

    To date there is no full-chain CCUS project involving offshore storage in China. This report gives a general review on major engi- neering requirements and their relevance and justification for offshore CO2 transportation and storage based on international experiences. The capital costs are mentioned briefly. The geological and system integration issues are expected to be covered in other reports.

  5. Efficient processing of TFO-directed psoralen DNA interstrand crosslinks by the UvrABC nuclease.

    Christensen, Laura A; Wang, Hong; Van Houten, Bennett; Vasquez, Karen M

    2008-12-01

    Photoreactive psoralens can form interstrand crosslinks (ICLs) in double-stranded DNA. In eubacteria, the endonuclease UvrABC plays a key role in processing psoralen ICLs. Psoralen-modified triplex-forming oligonucleotides (TFOs) can be used to direct ICLs to specific genomic sites. Previous studies of pyrimidine-rich methoxypsoralen-modified TFOs indicated that the TFO inhibits cleavage by UvrABC. Because different chemistries may alter the processing of TFO-directed ICLs, we investigated the effect of another type of triplex formed by purine-rich TFOs on the processing of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) ICLs by the UvrABC nuclease. Using an HMT-modified TFO to direct ICLs to a specific site, we found that UvrABC made incisions on the purine-rich strand of the duplex approximately 3 bases from the 3'-side and approximately 9 bases from the 5'-side of the ICL, within the TFO-binding region. In contrast to previous reports, the UvrABC nuclease cleaved the TFO-directed psoralen ICL with a greater efficiency than that of the psoralen ICL alone. Furthermore, the TFO was dissociated from its duplex binding site by UvrA and UvrB. As mutagenesis by TFO-directed ICLs requires nucleotide excision repair, the efficient processing of these lesions supports the use of triplex technology to direct DNA damage for genome modification. PMID:18996898

  6. WeltABC | kinderKAMERA

    Christian Schreger

    2013-09-01

    Full Text Available Das WeltABC ist eine Einladung zur Reise durch Sprache(n und damit verknüpften Bildern. Es ist eine Sammlung scheinbar klarer Begriffe, deren Ausgangspunkt die Sprache Deutsch ist. Diese Begriffe werden mit weiteren Begriffen assoziiert und durch Fotos illustriert.

  7. New Broom To Sweep Clean At ABC

    2007-01-01

    The Agricultural Bank of China (ABC) has a new president. The last non- publicly traded state-owned lender in China announced on July 6 that Xiang Junbo, former Vice Governor of the People’s Bank of China, or the central bank, had beenappointed to take o

  8. The H-loop in the Second Nucleotide-binding Domain of the Cystic Fibrosis Transmembrane Conductance Regulator is Required for Efficient Chloride Channel Closing

    Kloch, Monika; Milewski, Michał; Nurowska, Ewa; Dworakowska, Beata; Cutting, Garry R; Dołowy, Krzysztof

    2010-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that functions as a cAMP-activated chloride channel. The recent model of CFTR gating predicts that the ATP binding to both nucleotide-binding domains (NBD1 and NBD2) of CFTR is required for the opening of the channel, while the ATP hydrolysis at NBD2 induces subsequent channel closing. In most ABC proteins, efficient hydrolysis of ATP requires the presence of the invariant histidine res...

  9. THE IMPACT OF PROCUREMENT METHOD OF THE TRANSPORT SERVICES TO THE FINANCIAL REQUIREMENT OF PERFORMANCE CONTRACTING ENTITY

    Miloš POLIAK; Anna KRIŽANOVÁ; Štefánia SEMANOVÁ; Ľubica ŠTEFÁNIKOVÁ

    2013-01-01

    The paper deals with the issue of the impact of procurement method of the transport services to the financial requirement of performance contracting entity. In the first part, it describes the current state of transport services financing in the Slovak Republic and risk analysis, which is related to transport services procurement and conclusion of the service contract in the transport services ensuring. The rest of the paper defines the impact of procurement method of the transport services t...

  10. 40 CFR 80.350 - What alternative sulfur standards and requirements apply to importers who transport gasoline by...

    2010-07-01

    ... requirements apply to importers who transport gasoline by truck? 80.350 Section 80.350 Protection of... ADDITIVES Gasoline Sulfur Sampling, Testing and Retention Requirements for Refiners and Importers § 80.350 What alternative sulfur standards and requirements apply to importers who transport gasoline by...

  11. Capital requirements for the transportation of energy materials: 1979 ARC estimates. Final report

    1979-09-28

    TERA's estimates of capital requirements to transport natural gas, crude oil, petroleum products, and coal in the US by 1990 are presented. It is a continuation of a 1978 study (EAPA 5:3946) to perform a similar analysis on 1979 scenarios. Scenarios B, C, and D from the EIA's Mid-range Energy Forecasting Systems, as used in the 1979 Annual Report to Congress (ARC), were provided as a basis for the analysis and represent three alternative futures. Summaries of transportation investment requirements through 1990 are given for Scenarios B, C, and D. Total investment requirements for the three models (pipelines, railroads, waterways) and the three energy commodities (coal, petroleum, petroleum products, natural gas) are estimated to range between $35.3 and $42.7 billion by 1990 depending on the scenario.

  12. Baseline requirements of the proposed action for the Transportation Management Division routing models

    Johnson, P.E.; Joy, D.S.

    1995-02-01

    The potential impacts associated with the transportation of hazardous materials are important to shippers, carriers, and the general public. This is particularly true for shipments of radioactive material. The shippers are primarily concerned with safety, security, efficiency, and equipment requirements. The carriers are concerned with the potential impact that radioactive shipments may have on their operations--particularly if such materials are involved in an accident. The general public has also expressed concerns regarding the safety of transporting radioactive and other hazardous materials through their communities. Because transportation routes are a central concern in hazardous material transport, the prediction of likely routes is the first step toward resolution of these issues. In response to these routing needs, several models have been developed over the past fifteen years at Oak Ridge National Laboratory (ORNL). The HIGHWAY routing model is used to predict routes for truck transportation, the INTERLINE routing model is used to predict both rail and barge routes, and the AIRPORT locator model is used to determine airports with specified criteria near a specific location. As part of the ongoing improvement of the US Department of Energy`s (DOE) Environmental Management Transportation Management Division`s (EM-261) computer systems and development efforts, a Baseline Requirements Assessment Session on the HIGHWAY, INTERLINE, and AIRPORT models was held at ORNL on April 27, 1994. The purpose of this meeting was to discuss the existing capabilities of the models and data bases and to review enhancements of the models and data bases to expand their usefulness. The results of the Baseline Requirements Assessment Section will be discussed in this report. The discussions pertaining to the different models are contained in separate sections.

  13. Baseline requirements of the proposed action for the Transportation Management Division routing models

    The potential impacts associated with the transportation of hazardous materials are important to shippers, carriers, and the general public. This is particularly true for shipments of radioactive material. The shippers are primarily concerned with safety, security, efficiency, and equipment requirements. The carriers are concerned with the potential impact that radioactive shipments may have on their operations--particularly if such materials are involved in an accident. The general public has also expressed concerns regarding the safety of transporting radioactive and other hazardous materials through their communities. Because transportation routes are a central concern in hazardous material transport, the prediction of likely routes is the first step toward resolution of these issues. In response to these routing needs, several models have been developed over the past fifteen years at Oak Ridge National Laboratory (ORNL). The HIGHWAY routing model is used to predict routes for truck transportation, the INTERLINE routing model is used to predict both rail and barge routes, and the AIRPORT locator model is used to determine airports with specified criteria near a specific location. As part of the ongoing improvement of the US Department of Energy's (DOE) Environmental Management Transportation Management Division's (EM-261) computer systems and development efforts, a Baseline Requirements Assessment Session on the HIGHWAY, INTERLINE, and AIRPORT models was held at ORNL on April 27, 1994. The purpose of this meeting was to discuss the existing capabilities of the models and data bases and to review enhancements of the models and data bases to expand their usefulness. The results of the Baseline Requirements Assessment Section will be discussed in this report. The discussions pertaining to the different models are contained in separate sections

  14. An Electrically Tight In Vitro Blood-Brain Barrier Model Displays Net Brain-to-Blood Efflux of Substrates for the ABC Transporters, P-gp, Bcrp and Mrp-1

    Helms, Hans Christian; Hersom, Maria; Kuhlmann, Louise Borella;

    2014-01-01

    Efflux transporters of the ATP-binding cassette superfamily including breast cancer resistance protein (Bcrp/Abcg2), P-glycoprotein (P-gp/Abcb1) and multidrug resistance-associated proteins (Mrp's/Abcc's) are expressed in the blood-brain barrier (BBB). The aim of this study was to investigate if a......143, zosuquidar, reversan and MK 571 alone or in combinations. Digoxin was mainly transported via P-gp, estrone-3-sulphate via Bcrp and Mrp's and etoposide via P-gp and Mrp's. The expression of P-gp, Bcrp and Mrp-1 was confirmed using immunocytochemistry. The findings indicate that P-gp, Bcrp and at...... least one isoform of Mrp are functionally expressed in our bovine/rat co-culture model and that the model is suitable for investigations of small molecule transport....

  15. A note on ABC implementation: An evaluation case.

    Nur Naha Abu Mansor

    2004-12-01

    Full Text Available Whilst featuring prominently in management accounting research for at least the last decade, activity-based costing (ABC may have reached a plateau in term of implementation and success. This paper reports the results of a survey of activity-based costing (ABC in a telecommunication organization. The study considers ABC information and systems being used in various parts of division of the company. It reports on success measures adapted from the Foster and Swenson (1997 survey of ABC implementation.The Survey data was collected through a questionnaire to users of ABC sent through the company's e-mail system. 243 questionnaires were sent out for the 16 different ABC systems developed, 173 usable questionnaires were returned (a response rate of 71%. The respondents were selected based on their experience in using the various ABC systems irrespective of the divisions they operated in. They were mainly senior executives and executive level employees.The study is reported in four parts. Firstly, it reports on the usage of ABC information as compared to the traditional costing systems. It was found that budgeting, improving opportunities and aiding in product management decisions were highly scored as the main usage of ABC/M implementation. It states that ABC/M helped change in company's strategic focus, pricing strategy and changes in operations. Interestingly, it was found that ABC implementation provided almost no financial improvements, as perceived by respondents. Finally, the overall evaluation of the ABC systems is perceived by the users of the various ABC systems was to be mostly unsuccessful. A comparison of the survey results is made with the Foster and Swenson (1997 survey. The work is hoped to make a contribution to studying ABC implementation success in various settings.

  16. Analysis of operational requirements for medium density air transportation. Volume 1: Summary

    1975-01-01

    The medium density air travel market was studied to determine the aircraft design and operational requirements. The impact of operational characteristics on the air travel system and the economic viability of the study aircraft were also evaluated. Medium density is defined in terms of numbers of people transported (20 to 500 passengers per day on round trip routes), and frequency of service ( a minumium of two and maximum of eight round trips per day) for 10 regional carriers. The operational characteristics of aircraft best suited to serve the medium density air transportation market are determined and a basepoint aircraft is designed from which tradeoff studies and parametric variations could be conducted. The impact of selected aircraft on the medium density market, economics, and operations is ascertained. Research and technology objectives for future programs in medium density air transportation are identified and ranked.

  17. Application of the revised requirements of IAEA St-1 to the transport of natural uranium hexafluoride

    The 1996 issue of IAEA ST1 (formerly SS6) will, from january 2001, call for transport cylinders containing natural uranium hexafluoride ('hex') to be considered against the requirements of the IAEA thermal tests. In parallel with the revision of SS6, the IAEA established a coordinated research program (CRP) to consider the behaviour of hex cylinders in fires. BNFL has contributed to that CRP principally in the areas of computer modelling of the thermophysical processes occurring within the cylinder and of the mechanical behaviour of the cylinder itself. Confidence in the various computer simulations has grown steadily as the CRP progressed. With the benefit of results emerging from the Tenerife test program, our calculations now predict that an unprotected, fully loaded, 48Y hex cylinder would be expected to survive the thermal test. Since the expectation is that the unprotected fully loaded 48Y cylinders should (marginally) survive the fire test, and also taking account both of the excellent safety record to date of hex transports, and the rarity of intense long-lived fires in transport situations, there is clearly very little safety benefit to be gained from the use of fire protection covers on natural hex cylinders. However, the physical handling of such covers, necessarily large and cumbersome, and their relevant fastening, particularly during packaging, loading, unloading and un-packaging in transport depots would, for every cylinder movement, impose additional risks of injury to transport system operators, which we believe could outweigh the small and more hypothetical benefit of improved fire resistance while in transit. We therefore maintain that it would be counterproductive, in both overall safety and in cost terms, to require protective coverings on these cylinders for transport. Based on the estimates made to date, we believe that the unprotected 48 inch cylinders should continue to be the normal package in commercial use, under the relevant

  18. A method for determining the spent-fuel contribution to transport cask containment requirements

    This report examines containment requirements for spent-fuel transport containers that are transported under normal and hypothetical accident conditions. A methodology is described that estimates the probability of rod failure and the quantity of radioactive material released from breached rods. This methodology characterizes the dynamic environment of the cask and its contents and deterministically models the peak stresses that are induced in spent-fuel cladding by the mechanical and thermal dynamic environments. The peak stresses are evaluated in relation to probabilistic failure criteria for generated or preexisting ductile tearing and material fractures at cracks partially through the wall in fuel rods. Activity concentrations in the cask cavity are predicted from estimates of the fraction of gases, volatiles, and fuel fines that are released when the rod cladding is breached. Containment requirements based on the source term are calculated in terms of maximum permissible volumetric leak rates from the cask. Calculations are included for representative cask designs

  19. A method for determining the spent-fuel contribution to transport cask containment requirements

    Sanders, T.L.; Seager, K.D. [Sandia National Labs., Albuquerque, NM (United States); Rashid, Y.R.; Barrett, P.R. [ANATECH Research Corp., La Jolla, CA (United States); Malinauskas, A.P. [Oak Ridge National Lab., TN (United States); Einziger, R.E. [Pacific Northwest Lab., Richland, WA (United States); Jordan, H. [EG and G Rocky Flats, Inc., Golden, CO (United States). Rocky Flats Plant; Duffey, T.A.; Sutherland, S.H. [APTEK, Inc., Colorado Springs, CO (United States); Reardon, P.C. [GRAM, Inc., Albuquerque, NM (United States)

    1992-11-01

    This report examines containment requirements for spent-fuel transport containers that are transported under normal and hypothetical accident conditions. A methodology is described that estimates the probability of rod failure and the quantity of radioactive material released from breached rods. This methodology characterizes the dynamic environment of the cask and its contents and deterministically models the peak stresses that are induced in spent-fuel cladding by the mechanical and thermal dynamic environments. The peak stresses are evaluated in relation to probabilistic failure criteria for generated or preexisting ductile tearing and material fractures at cracks partially through the wall in fuel rods. Activity concentrations in the cask cavity are predicted from estimates of the fraction of gases, volatiles, and fuel fines that are released when the rod cladding is breached. Containment requirements based on the source term are calculated in terms of maximum permissible volumetric leak rates from the cask. Calculations are included for representative cask designs.

  20. Review of Transporter-Related Postmarketing Requirement or Postmarketing Commitment Studies.

    Fan, Ying; Sun, Bo; Agarwal, Sheetal; Zhang, Lei

    2016-07-01

    The objectives of this report are to summarize the content and status of transporter-related postmarketing requirement (PMR)/postmarketing commitment (PMC) studies in new drug applications (NDAs) approved by the U.S. Food and Drug Administration (FDA) and to discuss the reasons for requesting such studies and the impact of PMR/PMC study results on labeling to guide the optimal use of the drugs. Multiple data sources were searched to collect information on transporter-related PMR/PMC studies between January 1999 and May 2015. A total of 40 transporter-related PMR/PMC study requests were issued for 35 NDAs. Among these PMR/PMC studies, 27 requested studies related to P-glycoprotein. As of May 31, 2015, 34 transporter-related PMR/PMC studies (85%) are considered "fulfilled" (per the FDA's PMR/PMC website), and 22 (65%) resulted in labeling updates. The majority of the PMR/PMC studies are for drugs in the therapeutic areas of anti-infectives, oncology, and neurology. The results from PMR/PMC studies are important for dosing optimization and are often included in the updated labeling. Because a significant lag time is anticipated between drug approval and PMR/PMC fulfillment, NDA applicants are encouraged to include transporter-related assessments in clinical drug development programs for drug products. PMID:27385175

  1. Intracellular transport of secretory proteins in the pancreatic exocrine cell. IV. Metabolic requirements.

    Jamieson, J D; Palade, G E

    1968-12-01

    Since in the pancreatic exocrine cell synthesis and intracellular transport of secretory proteins can be uncoupled (1), it is possible to examine separately the metabolic requirements of the latter process. To this intent, guinea pig pancreatic slices were pulse labeled with leucine-(3)H for 3 min and incubated post-pulse for 37 min in chase medium containing 5 x 10(-4)M cycloheximide and inhibitors of glycolysis, respiration, or oxidative phosphorylation. In each case, the effect on transport was assessed by measuring the amount of labeled secretory proteins found in zymogen granule fractions isolated from the corresponding slices. This assay is actually a measure of the efficiency of transport of secretory proteins from the cisternae of the rough endoplasmic reticulum (RER) to the condensing vacuoles of the Golgi complex which are recovered in the zymogen granule fraction (16). The results indicate that transport is insensitive to glycolytic inhibitors (fluoride, iodoacetate) but is blocked by respiratory inhibitors (N(2), cyanide, Antimycin A) and by inhibitors of oxidative phosphorylation (dinitrophenol, oligomycin). Except for Antimycin A, the effect is reversible. Parallel radioautographic studies and cell fractionation procedures applied to microsomal subfractions have indicated that the energy-dependent step is located between the transitional elements of the RER and the small, smooth-surfaced vesicles at the periphery of the Golgi complex. Radiorespirometric data indicate that the substrates oxidized to support transport are endogenous long-chain fatty acids. PMID:5699933

  2. Negative membrane curvature catalyzes nucleation of endosomal sorting complex required for transport (ESCRT)-III assembly

    Lee, IH; Kai, H; Carlson, LA; Groves, JT; Hurley, JH

    2015-01-01

    The endosomal sorting complexes required for transport (ESCRT) machinery functions in HIV-1 budding, cytokinesis, multivesicular body biogenesis, and other pathways, in the course of which it interacts with concave membrane necks and bud rims. To test the role of membrane shape in regulating ESCRT assembly, we nanofabricated templates for invaginated supported lipid bilayers. The assembly of the core ESCRT-III subunit CHMP4B/Snf7 is preferentially nucleated in the resulting 100-nm-deep membra...

  3. Nanostructured assemblies from amphiphilic ABC multiblock polymers

    Hillmyer, Marc A.

    2012-02-01

    Amphiphilic AB diblock copolymers containing a water compatible segment can self-assemble in aqueous media to give supramolecular structures that include simple spherical micelles and macromolecular vesicles termed polymersomes. Amphiphilic ABA triblocks with hydrophobic end blocks can adopt analogous structures but can also form gels at high polymer concentrations. The structural and chemical diversity demonstrated in block copolymer micelles and gels makes them attractive for applications ranging from drug delivery to personal care products to nanoreactors. The inclusion of a third block in amphiphilic ABC triblock systems can lead to a much wider array of self-assembled structures that depend not only on composition but also on block sequence, architecture and incompatibility considerations. I will present our recent efforts on tuning micelle and gel structure and behavior using controlled architecture ABC triblocks. The combination of diverse polymer segments into a single macromolecule is a powerful method for development of self-assembled structures with both new form and new function.

  4. MANFAAT DATA WAREHOUSE PADA PT ABC

    Evaristus Didik Madyatmadja

    2013-11-01

    Full Text Available The purpose of this paper is to analyze the current system to find out problems faced by the company and propose alternative solutions and generate information needed by management by designing a data warehouse according to the information needs of PT ABC. This research implements analysis and design of data warehouse by Ralph Kimball and Ross cited by Connolly and Begg, known as Nine-Step Methodology. The result obtained is a data warehouse application that may present a multidimensional historical data that can assist the management in decisions. Designing data warehouse at PT ABC makes concise the enterprise data and can be viewed from several dimensions. It helps users analyze data for strategic decision quickly and accurately.

  5. Harpunsoiton ABC : harpunsoiton alkeiskirjan luominen lapselle

    Purhonen, Tiia

    2015-01-01

    Tässä opinnäytetyössä kerron tekeillä olevan projektini Harpunsoiton ABC:n luomisen taustoista ja vaiheista. Projektin tavoitteena on valmistaa lapsille suunnattu harpunsoiton alkeiskirja. Opinnäyteyössäni etenen harpunsoiton harrastamisen historiasta edelleen nykytilanteeseen Suomessa ja esittelen modernin harpun ominaisuuksia ja soittotekniikkaa. Vertailen nykyistä harpun opetusmateriaalin kirjoa muiden soittimien oppikirjojen laajaan tarjontaan etsien hyviä pedagogisia ideoita. Ope...

  6. ABC of physics a very brief guide

    Okun, Lev Borisovich

    2012-01-01

    This little book concentrates on the foundations of modern physics (its "ABC's") and its most fundamental constants: c - the velocity of light and ? - the quantum of action. First of all, the book is addressed to professional physicists, but in order to achieve maximal concentration and clarity it uses the simplest (high school) mathematics. As a result many pages of the book will be useful to college students and may appeal to a more general audience.

  7. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    Liu, Shumin; Zhou, Shun; Tian, Ling; Guo, Enen; Luan, Yunxia; Zhang, Jianzhen; Li, Sheng

    2011-01-01

    Background The ATP-binding cassette (ABC) transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight subfamil...

  8. Caracterización y análisis del coste de mejora de la calidad de servicio en transporte urbano en autobús, basado en la metodología ABC (activity based costing)

    García Pastor, Antonio

    2016-01-01

    El estudio de la calidad de servicio en transporte público ha tenido distintos enfoques, tanto en la investigación como en sus aplicaciones prácticas, tales como el desarrollo del método SERVQUAL, los proyectos QUATTRO y EQUIP de la Comisión Europea, el Transit Capacity and Quality of Service Manual y finalmente el estándar EN 13816, siendo la consideración principal la calidad desde el punto de vista del usuario, es decir, la calidad percibida, cuyo objetivo en la práctica tra...

  9. Phase behavior of model ABC triblock copolymers

    Chatterjee, Joon

    The phase behavior of poly(isoprene-b-styrene- b-ethylene oxide) (ISO), a model ABC triblock copolymer has been studied. This class of materials exhibit self-assembly, forming a large array of ordered morphologies at length scales of 5-100 nm. The formation of stable three-dimensionally continuous network morphologies is of special interest in this study. Since these nanostructures considerably impact the material properties, fundamental knowledge for designing ABC systems have high technological importance for realizing applications in the areas of nanofabrication, nanoporous media, separation membranes, drug delivery and high surface area catalysts. A comprehensive framework was developed to describe the phase behavior of the ISO triblock copolymers at weak to intermediate segregation strengths spanning a wide range of composition. Phases were characterized through a combination of characterization techniques, including small angle x-ray scattering, dynamic mechanical spectroscopy, transmission electron microscopy, and birefringence measurements. Combined with previous investigations on ISO, six different stable ordered state symmetries have been identified: lamellae (LAM), Fddd orthorhombic network (O70), double gyroid (Q230), alternating gyroid (Q214), hexagonal (HEX), and body-centered cubic (BCC). The phase map was found to be somewhat asymmetric around the fI = fO isopleth. This work provides a guide for theoretical studies and gives insight into the intricate effects of various parameters on the self-assembly of ABC triblock copolymers. Experimental SAXS data evaluated with a simple scattering intensity model show that local mixing varies continuously across the phase map between states of two- and three-domain segregation. Strategies of blending homopolymers with ISO triblock copolymer were employed for studying the swelling properties of a lamellar state. Results demonstrate that lamellar domains swell or shrink depending upon the type of homopolymer that

  10. The ATP-binding Cassette Transporter OsABCG15 is Required for Anther Development and Pollen Fertility in Rice

    Bai-Xiao Niu; Fu-Rong He; Ming He; Ding Ren; Le-Tian Chen; Yao-Guang Liu

    2013-01-01

    Plant male reproductive development is a complex biological process,but the underlying mechanism is not well understood.Here,we characterized a rice (Oryza sativa L.) male sterile mutant.Based on mapbased cloning and sequence analysis,we identified a 1,459-bp deletion in an adenosine triphosphate (ATP)-binding cassette (ABC) transporter gene,OsABCG15,causing abnormal anthers and male sterility.Therefore,we named this mutant osabcg15.Expression analysis showed that OsABCG15 is expressed specifically in developmental anthers from stage 8 (meiosis Ⅱ stage) to stage 10 (late microspore stage).Two genes CYP704B2 and WDA1,involved in the biosynthesis of very-long-chain fatty acids for the establishment of the anther cuticle and pollen exine,were downregulated in osabcg15 mutant,suggesting that OsABCG15 may play a key function in the processes related to sporopollenin biosynthesis or sporopollenin transfer from tapetal cells to anther locules.Consistently,histological analysis showed that osabcg15 mutants developed obvious abnormality in postmeiotic tapetum degeneration,leading to rapid degredation of young microspores.The results suggest that OsABCG15 plays a critical role in exine formation and pollen development,similar to the homologous gene of AtABCG26 in Arabidopsis.This work is helpful to understand the regulatory network in rice anther development.

  11. Structural basis for allosteric cross-talk between the asymmetric nucleotide binding sites of a heterodimeric ABC exporter.

    Hohl, Michael; Hürlimann, Lea M; Böhm, Simon; Schöppe, Jendrik; Grütter, Markus G; Bordignon, Enrica; Seeger, Markus A

    2014-07-29

    ATP binding cassette (ABC) transporters mediate vital transport processes in every living cell. ATP hydrolysis, which fuels transport, displays positive cooperativity in numerous ABC transporters. In particular, heterodimeric ABC exporters exhibit pronounced allosteric coupling between a catalytically impaired degenerate site, where nucleotides bind tightly, and a consensus site, at which ATP is hydrolyzed in every transport cycle. Whereas the functional phenomenon of cooperativity is well described, its structural basis remains poorly understood. Here, we present the apo structure of the heterodimeric ABC exporter TM287/288 and compare it to the previously solved structure with adenosine 5'-(β,γ-imido)triphosphate (AMP-PNP) bound at the degenerate site. In contrast to other ABC exporter structures, the nucleotide binding domains (NBDs) of TM287/288 remain in molecular contact even in the absence of nucleotides, and the arrangement of the transmembrane domains (TMDs) is not influenced by AMP-PNP binding, a notion confirmed by double electron-electron resonance (DEER) measurements. Nucleotide binding at the degenerate site results in structural rearrangements, which are transmitted to the consensus site via two D-loops located at the NBD interface. These loops owe their name from a highly conserved aspartate and are directly connected to the catalytically important Walker B motif. The D-loop at the degenerate site ties the NBDs together even in the absence of nucleotides and substitution of its aspartate by alanine is well-tolerated. By contrast, the D-loop of the consensus site is flexible and the aspartate to alanine mutation and conformational restriction by cross-linking strongly reduces ATP hydrolysis and substrate transport. PMID:25030449

  12. Competence and Capacity-Building Requirements in Transport and Logistics Market

    Bazaras Darius

    2016-03-01

    Full Text Available The article analyses theoretical aspects of the qualitative assessment of logistics and transport specialists’ potential, as well as provides research results on competence and capacity-building process in preparation of logistics and transport specialists. The results enabled to identify problematic areas of graduate integration in the labour market and foresee the possible requirements for professional development. In some cases, internal motivation is related to problematic points or negative view from the environment, as well as poor understanding of knowledge benefits in the person’s practical life. Training and education is objectively at the meeting point between research, technological and organisational systems. In such conditions the objective of the education on logistics is to offer the customers the best possible service in the field of the transfer of knowledge in harmony with the trend in logistics programme development. The main problems and perspectives of logistics education and training process are discussed in this article.

  13. LSSVM-ABC Algorithm for Stock Price prediction

    Hegazy, Osman; Soliman, Omar S.; Salam, Mustafa Abdul

    2014-01-01

    In this paper, Artificial Bee Colony (ABC) algorithm which inspired from the behavior of honey bees swarm is presented. ABC is a stochastic population-based evolutionary algorithm for problem solving. ABC algorithm, which is considered one of the most recently swarm intelligent techniques, is proposed to optimize least square support vector machine (LSSVM) to predict the daily stock prices. The proposed model is based on the study of stocks historical data, technical indicators and optimizing...

  14. Television Viewing and ABC Program Policy: An Econometric Study

    Glenn Withers

    1985-01-01

    There has been a long-standing debate over whether the ABC should engage in programming that is directly competitive with commercial television stations. This study uses an econometric model of television viewing to examine determinants of television set ownership and use, of ABC viewing levels and of diversity in programming. It is established that complementary not competitive programming most enhances ABC viewing levels, though production of greater diversity of television programming may ...

  15. Capital requirements for the transportation of energy materials: 1979 ARC estimates. Draft final report

    1980-08-13

    This report contains TERA's estimates of capital requirements to transport natural gas, crude oil, petroleum products, and coal in the United States by 1990. The low, medium, and high world-oil-price scenarios from the EIA's Mid-range Energy Forecasting System (MEFS), as used in the 1979 Annual Report to Congress (ARC), were provided as a basis for the analysis and represent three alternative futures. TERA's approach varies by energy commodity to make best use of the information and analytical tools available. Summaries of transportation investment requirements through 1990 are given. Total investment requirements for three modes (pipelines, rails, waterways and the three energy commodities can accumulate to a $49.9 to $50.9 billion range depending on the scenario. The scenarios are distinguished primarily by the world price of oil which, given deregulation of domestic oil prices, affects US oil prices even more profoundly than in the past. The high price of oil, following the evidence of the last year, is projected to hold demand for oil below the recent past.

  16. Biliary cholesterol secretion: More than a simple ABC

    Arne Dikkers, Uwe JF Tietge

    2010-12-01

    Full Text Available Biliary cholesterol secretion is a process important for 2 major disease complexes, atherosclerotic cardiovascular disease and cholesterol gallstone disease. With respect to cardiovascular disease, biliary cholesterol secretion is regarded as the final step for the elimination of cholesterol originating from cholesterol-laden macrophage foam cells in the vessel wall in a pathway named reverse cholesterol transport. On the other hand, cholesterol hypersecretion into the bile is considered the main pathophysiological determinant of cholesterol gallstone formation. This review summarizes current knowledge on the origins of cholesterol secreted into the bile as well as the relevant processes and transporters involved. Next to the established ATP-binding cassette (ABC transporters mediating the biliary secretion of bile acids (ABCB11, phospholipids (ABCB4 and cholesterol (ABCG5/G8, special attention is given to emerging proteins that modulate or mediate biliary cholesterol secretion. In this regard, the potential impact of the phosphatidylserine flippase ATPase class I type 8B member 1, the Niemann Pick C1-like protein 1 that mediates cholesterol absorption and the high density lipoprotein cholesterol uptake receptor, scavenger receptor class B type I, is discussed.

  17. Concepts for Life Cycle Cost Control Required to Achieve Space Transportation Affordability and Sustainability

    Rhodes, Russel E.; Zapata, Edgar; Levack, Daniel J. H.; Robinson, John W.; Donahue, Benjamin B.

    2009-01-01

    Cost control must be implemented through the establishment of requirements and controlled continually by managing to these requirements. Cost control of the non-recurring side of life cycle cost has traditionally been implemented in both commercial and government programs. The government uses the budget process to implement this control. The commercial approach is to use a similar process of allocating the non-recurring cost to major elements of the program. This type of control generally manages through a work breakdown structure (WBS) by defining the major elements of the program. If the cost control is to be applied across the entire program life cycle cost (LCC), the approach must be addressed very differently. A functional breakdown structure (FBS) is defined and recommended. Use of a FBS provides the visibifity to allow the choice of an integrated solution reducing the cost of providing many different elements of like function. The different functional solutions that drive the hardware logistics, quantity of documentation, operational labor, reliability and maintainability balance, and total integration of the entire system from DDT&E through the life of the program must be fully defined, compared, and final decisions made among these competing solutions. The major drivers of recurring cost have been identified and are presented and discussed. The LCC requirements must be established and flowed down to provide control of LCC. This LCC control will require a structured rigid process similar to the one traditionally used to control weight/performance for space transportation systems throughout the entire program. It has been demonstrated over the last 30 years that without a firm requirement and methodically structured cost control, it is unlikely that affordable and sustainable space transportation system LCC will be achieved.

  18. Influence of target requirements on the production, acceleration, transport, and focusing of ion beams

    Bangerter, R.O.; Mark, J.W.K.; Meeker, D.J.; Judd, D.L.

    1981-01-01

    We have calculated the energy gain of ion-driven fusion targets as a function of input energy, ion range, and focal spot radius. For heavy-ion drivers a given target gain, together with final-lens properties, determines a 6-D phase space volume which must exceed that occupied by the ion beam. Because of Liouville's theorem and the inevitability of some phase space dilutions, the beams's 6-D volume will increase between the ion source and the target. This imposes important requirements on accelerators and on transport and focusing systems.

  19. Influence of target requirements on the production, acceleration, transport, and focusing of ion beams

    We have calculated the energy gain of ion-driven fusion targets as a function of input energy, ion range, and focal spot radius. For heavy-ion drivers a given target gain, together with final-lens properties, determines a 6-D phase space volume which must exceed that occupied by the ion beam. Because of Liouville's theorem and the inevitability of some phase space dilutions, the beams's 6-D volume will increase between the ion source and the target. This imposes important requirements on accelerators and on transport and focusing systems

  20. Effect of the deletion of qmoABC and the promoter distal gene encoding a hypothetical protein on sulfate-reduction in Desulfovibrio vulgaris Hildenborough

    Zane, Grant M.; Yen, Huei-chi Bill; Wall, Judy D.

    2010-03-18

    The pathway of electrons required for the reduction of sulfate in sulfate-reducing bacteria (SRB) is not yet fully characterized. In order to determine the role of a transmembrane protein complex suggested to be involved in this process, a deletion of Desulfovibrio vulgaris Hildenborough was created by marker exchange mutagenesis that eliminated four genes putatively encoding the QmoABC complex and a hypothetical protein (DVU0851). The Qmo complex (quinone-interacting membrane-bound oxidoreductase) is proposed to be responsible for transporting electrons to the dissimilatory adenosine-5?phosphosulfate (APS) reductase in SRB. In support of the predicted role of this complex, the deletion mutant was unable to grow using sulfate as its sole electron acceptor with a range of electron donors. To explore a possible role for the hypothetical protein in sulfate reduction, a second mutant was constructed that had lost only the gene that codes for DVU0851. The second constructed mutant grew with sulfate as the sole electron acceptor; however, there was a lag that was not present with the wild-type or complemented strain. Neither deletion strain was significantly impaired for growth with sulfite or thiosulfate as terminal electron acceptor. Complementation of the D(qmoABC-DVU0851) mutant with all four genes or only the qmoABC genes restored its ability to grow by sulfate respiration. These results confirmed the prediction that the Qmo complex is in the electron pathway for sulfate-reduction and revealed that no other transmembrane complex could compensate when Qmo was lacking.

  1. Serum albumin promotes ATP-binding cassette transporter-dependent sterol uptake in yeast

    Marek, Magdalena; Silvestro, Daniele; Fredslund, Maria D.;

    2014-01-01

    Sterol uptake in fungi is a multistep process that involves interaction between external sterols and the cell wall, incorporation of sterol molecules into the plasma membrane, and subsequent integration into intracellular membranes for turnover. ATP-binding cassette (ABC) transporters have been i...... donor in ATP-binding cassette-transporter-dependent sterol uptake, a process potentially important for growth of Candida glabrata inside infected humans....... implicated in sterol uptake, but key features of their activity remain to be elucidated. Here, we apply fluorescent cholesterol (NBD-cholesterol) to monitor sterol uptake under anaerobic and aerobic conditions in two fungal species, Candida glabrata (Cg) and Saccharomyces cerevisiae (Sc). We found that in....... cerevisiae requires the presence of serum or albumin for efficient cholesterol uptake. These results suggest that albumin can serve as sterol donor in ABC transporter-dependent sterol uptake, a process potentially important for growth of C. glabrata inside infected humans. Serum albumin can serve as sterol...

  2. Assessment and Requirements of Nuclear Reaction Databases for GCR Transport in the Atmosphere and Structures

    Cucinotta, F. A.; Wilson, J. W.; Shinn, J. L.; Tripathi, R. K.

    1998-01-01

    The transport properties of galactic cosmic rays (GCR) in the atmosphere, material structures, and human body (self-shielding) am of interest in risk assessment for supersonic and subsonic aircraft and for space travel in low-Earth orbit and on interplanetary missions. Nuclear reactions, such as knockout and fragmentation, present large modifications of particle type and energies of the galactic cosmic rays in penetrating materials. We make an assessment of the current nuclear reaction models and improvements in these model for developing required transport code data bases. A new fragmentation data base (QMSFRG) based on microscopic models is compared to the NUCFRG2 model and implications for shield assessment made using the HZETRN radiation transport code. For deep penetration problems, the build-up of light particles, such as nucleons, light clusters and mesons from nuclear reactions in conjunction with the absorption of the heavy ions, leads to the dominance of the charge Z = 0, 1, and 2 hadrons in the exposures at large penetration depths. Light particles are produced through nuclear or cluster knockout and in evaporation events with characteristically distinct spectra which play unique roles in the build-up of secondary radiation's in shielding. We describe models of light particle production in nucleon and heavy ion induced reactions and make an assessment of the importance of light particle multiplicity and spectral parameters in these exposures.

  3. Use of simple transport equations to estimate waste package performance requirements

    A method of developing waste package performance requirements for specific nuclides is described. The method is based on: Federal regulations concerning permissible concentrations in solution at the point of discharge to the accessible environment; a simple and conservative transport model; baseline and potential worst-case release scenarios. Use of the transport model enables calculation of maximum permissible release rates within a repository in basalt for each of the scenarios. The maximum permissible release rates correspond to performance requirements for the engineered barrier system. The repository was assumed to be constructed in a basalt layer. For the cases considered, including a well drilled into an aquifer 1750 m from the repository center, little significant advantage is obtained from a 1000-yr as opposed to a 100-yr waste package. A 1000-yr waste package is of importance only for nuclides with half-lives much less than 100 yr which travel to the accessible environment in much less than 1000 yr. Such short travel times are extremely unlikely for a mined repository. Among the actinides, the most stringent maximum permissible release rates are for 236U and 234U. A simple solubility calculation suggests, however, that these performance requirements can be readily met by the engineered barrier system. Under the reducing conditions likely to occur in a repository located in basalt, uranium would be sufficiently insoluble that no solution could contain more than about 0.01% of the maximum permissible concentration at saturation. The performance requirements derived from the one-dimensional modeling approach are conservative by at least one to two orders of magnitude. More quantitative three-dimensional modeling at specific sites should enable relaxation of the performance criteria derived in this study. 12 references, 8 figures, 8 tables

  4. OSTOTOIMINNAN TEHOSTAMINEN : Case ABC Kokkolan ravintola

    Huldén, Elin

    2014-01-01

    Tämän toiminnallisen opinnäytetyön tarkoituksena oli kehittää ravintolan tilaus-toimitusrytmiä, huomioiden riviteho, logistiikka, ekologisuus ja varaston hallinta. Työn toimeksiantaja oli Keskipohjanmaan Osuuskauppa KPO, ja tutkimus kohdistui ABC Kokkola liikennemyymälän ravintolaan. Teoriaosa koostuu työn keskeisempien käsitteiden määrittelystä, kuten tilaus-toimitusketju, riviteho, varastonhallinta ja varastonkierto. Teoriaosuus sisältää myös toimeksiantajan esittelyn sekä tietoa toimin...

  5. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    Zhang Jianzhen; Luan Yunxia; Guo Enen; Tian Ling; Zhou Shun; Liu Shumin; Li Sheng

    2011-01-01

    Abstract Background The ATP-binding cassette (ABC) transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight...

  6. Dashboard auditing of ABC (Activity-Based Costing). Theoretical approaches

    Căpuşneanu, Sorinel/I

    2009-01-01

    This article aims to define the dashboard auditing according to the specifics of Activity-Based Costing method (ABC). It describes the main objectives of dashboard auditing, the criteria that a dashboard auditor should meet and the step-by-step stages of the entire dashboard auditing process of an enterprise from steel industry according to the Activity-Based Costing method (ABC).

  7. Dashboard Auditing of Activity-Based Costing (ABC)

    Sorinel Capusneanu

    2009-01-01

    This article aims to define the dashboard auditing according to the specifics of Activity-Based Costing method (ABC). It describes the main objectives of dashboard auditing, the criteria that a dashboard auditor should meet and the step-by-step stages of the entire dashboard auditing process according to the Activity-Based Costing method (ABC).

  8. Dashboard Auditing of Activity-Based Costing (ABC

    Sorinel Capusneanu

    2009-03-01

    Full Text Available This article aims to define the dashboard auditing according to the specifics of Activity-Based Costing method (ABC. It describes the main objectives of dashboard auditing, the criteria that a dashboard auditor should meet and the step-by-step stages of the entire dashboard auditing process according to the Activity-Based Costing method (ABC.

  9. Seasonal Expression of the Picocyanobacterial Phosphonate Transporter Gene phnD in the Sargasso Sea

    GeorgeSBullerjahn; RobCondon

    2010-01-01

    In phosphorus-limited marine environments, picocyanobacteria (Synechococcus and Prochlorococcus spp.) can hydrolyze naturally occurring phosphonates as a P source. Utilization of 2-aminoethylphosphonate (2-AEP) is dependent on expression of the phn genes, encoding functions required for uptake and C-P bond cleavage. Prior work has indicated that expression of picocyanobacterial phnD, encoding the phosphonate binding protein of the phosphonate ABC transporter, is a proxy for the assimilation...

  10. Sustained Delivery of Chondroitinase ABC from Hydrogel System

    Filippo Rossi

    2012-03-01

    Full Text Available In the injured spinal cord, chondroitin sulfate proteoglycans (CSPGs are the principal responsible of axon growth inhibition and they contribute to regenerative failure, promoting glial scar formation. Chondroitinase ABC (chABC is known for being able to digest proteoglycans, thus degrading glial scar and favoring axonal regrowth. However, its classic administration is invasive, infection-prone and clinically problematic. An agarose-carbomer (AC1 hydrogel, already used in SCI repair strategies, was here investigated as a delivery system capable of an effective chABC administration: the material ability to include chABC within its pores and the possibility to be injected into the target tissue were firstly proved. Subsequently, release kinetic and the maintenance of enzymatic activity were positively assessed: AC1 hydrogel was thus confirmed to be a feasible tool for chABC delivery and a promising device for spinal cord injury topic repair strategies.

  11. Transition fibre protein FBF1 is required for the ciliary entry of assembled intraflagellar transport complexes.

    Wei, Qing; Xu, Qingwen; Zhang, Yuxia; Li, Yujie; Zhang, Qing; Hu, Zeng; Harris, Peter C; Torres, Vicente E; Ling, Kun; Hu, Jinghua

    2013-01-01

    Sensory organelle cilia have critical roles in mammalian embryonic development and tissue homeostasis. Intraflagellar transport (IFT) machinery is required for the assembly and maintenance of cilia. Yet, how this large complex passes through the size-dependent barrier at the ciliary base remains enigmatic. Here we report that FBF1, a highly conserved transition fibre protein, is required for the ciliary import of assembled IFT particles at the ciliary base. We cloned dyf-19, the Caenorhabditis elegans homologue of human FBF1, in a whole-genome screen for ciliogenesis mutants. DYF-19 localizes specifically to transition fibres and interacts directly with the IFT-B component DYF-11/IFT54. Although not a structural component of transition fibres, DYF-19 is required for the transit of assembled IFT particles through the ciliary base. Furthermore, we found that human FBF1 shares conserved localization and function with its worm counterpart. We conclude that FBF1 is a key functional transition fibre component that actively facilitates the ciliary entry of assembled IFT machinery. PMID:24231678

  12. Automated container transportation using self-guided vehicles: Fernald site requirements

    A new opportunity to improve the safety and efficiency of environmental restoration operations, using robotics has emerged from advances in industry, academia, and government labs. Self-Guided Vehicles (SGV's) have recently been developed in industry and early systems have already demonstrated much, though not all, of the functionality necessary to support driverless transportation of waste within and between processing facilities. Improved materials databases are being developed by at least two DOE remediation sites, the Fernald Environmental Management Project (FEME) in the State of Ohio and the Hanford Complex in the State of Washington. SGV's can be developed that take advantage of the information in these databases and yield improved dispatch, waste tracking, report and shipment documentation. In addition, they will reduce the radiation hazard to workers and the risk of damaging containers through accidental collision. In this document, features of remediation sites that dictate the design of both the individual SGV's and the collective system of SGV's are presented, through the example of the site requirements at Fernald. Some concepts borrowed from the world of manufacturing are explained and then used to develop an integrated, holistic view of the remediation site as a pseudo-factory. Transportation methods at Fernald and anticipated growth in transport demand are analyzed. The new site-wide database under development at Fernald is presented so that advantageous and synergistic links between SGV's and information systems can be analyzed. Details of the SGV development proposed are submitted, and some results of a recently completed state of the art survey for SGV use in this application are also presented

  13. Automated container transportation using self-guided vehicles: Fernald site requirements

    Hazen, F.B.

    1993-09-01

    A new opportunity to improve the safety and efficiency of environmental restoration operations, using robotics has emerged from advances in industry, academia, and government labs. Self-Guided Vehicles (SGV`s) have recently been developed in industry and early systems have already demonstrated much, though not all, of the functionality necessary to support driverless transportation of waste within and between processing facilities. Improved materials databases are being developed by at least two DOE remediation sites, the Fernald Environmental Management Project (FEME) in the State of Ohio and the Hanford Complex in the State of Washington. SGV`s can be developed that take advantage of the information in these databases and yield improved dispatch, waste tracking, report and shipment documentation. In addition, they will reduce the radiation hazard to workers and the risk of damaging containers through accidental collision. In this document, features of remediation sites that dictate the design of both the individual SGV`s and the collective system of SGV`s are presented, through the example of the site requirements at Fernald. Some concepts borrowed from the world of manufacturing are explained and then used to develop an integrated, holistic view of the remediation site as a pseudo-factory. Transportation methods at Fernald and anticipated growth in transport demand are analyzed. The new site-wide database under development at Fernald is presented so that advantageous and synergistic links between SGV`s and information systems can be analyzed. Details of the SGV development proposed are submitted, and some results of a recently completed state of the art survey for SGV use in this application are also presented.

  14. Feasibility of the use of the Active Breathing Co ordinatorTM (ABC) in patients receiving radical radiotherapy for non-small cell lung cancer (NSCLC)

    Introduction: One method to overcome the problem of lung tumour movement in patients treated with radiotherapy is to restrict tumour motion with an active breathing control (ABC) device. This study evaluated the feasibility of using ABC in patients receiving radical radiotherapy for non-small cell lung cancer. Methods: Eighteen patients, median (range) age of 66 (44-82) years, consented to the study. A training session was conducted to establish the patient's breath hold level and breath hold time. Three planning scans were acquired using the ABC device. Reproducibility of breath hold was assessed by comparing lung volumes measured from the planning scans and the volume recorded by ABC. Patients were treated with a 3-field coplanar beam arrangement and treatment time (patient on and off the bed) and number of breath holds recorded. The tolerability of the device was assessed by weekly questionnaire. Quality assurance was performed on the two ABC devices used. Results: 17/18 patients completed 32 fractions of radiotherapy using ABC. All patients tolerated a maximum breath hold time >15 s. The mean (SD) patient training time was 13.8 (4.8) min and no patient found the ABC very uncomfortable. Six to thirteen breath holds of 10-14 s were required per session. The mean treatment time was 15.8 min (5.8 min). The breath hold volumes were reproducible during treatment and also between the two ABC devices. Conclusion: The use of ABC in patients receiving radical radiotherapy for NSCLC is feasible. It was not possible to predict a patient's ability to hold breath. A minimum tolerated breath hold time of 15 s is recommended prior to commencing treatment.

  15. The Role of the Actin Cytoskeleton and Lipid Rafts in the Localization and Function of the ABCC1 Transporter

    Jan Willem Kok

    2014-01-01

    Full Text Available ATP-binding cassette (ABC transporters are known to be important factors in multidrug resistance of tumor cells. Lipid rafts have been implicated in their localization in the plasma membrane, where they function as drug efflux pumps. This specific localization in rafts may support the activity of ABC/Abc transporters. This raises questions regarding the nature and composition of the lipid rafts that harbor ABC/Abc transporters and the dependence of ABC/Abc transporters—concerning their localization and activity—on lipid raft constituents. Here we review our work of the past 10 years aimed at evaluating whether ABC/Abc transporters are dependent on a particular membrane environment for their function. What is the nature of this membrane environment and which of the lipid raft constituents are important for this dependency? It turns out that cortical actin is of major importance for stabilizing the localization and function of the ABC/Abc transporter, provided it is localized in an actin-dependent subtype of lipid rafts, as is the case for human ABCC1/multidrug resistance-related protein 1 (MRP1 and rodent Abcc1/Mrp1 but not human ABCB1/P-glycoprotein (PGP. On the other hand, sphingolipids do not appear to be modulators of ABCC1/MRP1 (or Abcc1/Mrp1, even though they are coregulated during drug resistance development.

  16. The boron efflux transporter ROTTEN EAR is required for maize inflorescence development and fertility.

    Chatterjee, Mithu; Tabi, Zara; Galli, Mary; Malcomber, Simon; Buck, Amy; Muszynski, Michael; Gallavotti, Andrea

    2014-07-01

    Although boron has a relatively low natural abundance, it is an essential plant micronutrient. Boron deficiencies cause major crop losses in several areas of the world, affecting reproduction and yield in diverse plant species. Despite the importance of boron in crop productivity, surprisingly little is known about its effects on developing reproductive organs. We isolated a maize (Zea mays) mutant, called rotten ear (rte), that shows distinct defects in vegetative and reproductive development, eventually causing widespread sterility in its inflorescences, the tassel and the ear. Positional cloning revealed that rte encodes a membrane-localized boron efflux transporter, co-orthologous to the Arabidopsis thaliana BOR1 protein. Depending on the availability of boron in the soil, rte plants show a wide range of phenotypic defects that can be fully rescued by supplementing the soil with exogenous boric acid, indicating that rte is crucial for boron transport into aerial tissues. rte is expressed in cells surrounding the xylem in both vegetative and reproductive tissues and is required for meristem activity and organ development. We show that low boron supply to the inflorescences results in widespread defects in cell and cell wall integrity, highlighting the structural importance of boron in the formation of fully fertile reproductive organs. PMID:25035400

  17. Comparative economics for DUCRETE spent fuel storage cask handling, transportation, and capital requirements

    This report summarizes economic differences between a DUCRETE spent nuclear fuel storage cask and a conventional concrete storage cask in the areas of handling, transportation, and capital requirements. The DUCRETE cask is under evaluation as a new technology that could substantially reduce the overall costs of spent fuel and depleted U disposal. DUCRETE incorporates depleted U in a Portland cement mixture and functions as the cask's primary radiation barrier. The cask system design includes insertion of the US DOE Multi-Purpose Canister inside the DUCRETE cask. The economic comparison is from the time a cask is loaded in a spent fuel pool until it is placed in the repository and includes the utility and overall US system perspectives

  18. Schedules of requirements for the transport of specified types of radioactive material consignments (as amended 1990)

    This publication replaces the previous issue of Safety Series No. 80. It includes the changes to Safety Series No. 80 contained in Supplement 1988 to the Regulations for the Safe Transport of Radioactive Material as well as some modifications adopted by the Review Panel meeting which convened in Vienna, from 10 to 14 July 1989. The old IAEA Safety Series style is maintained for the convenience of the user, although the old style has now been superseded by a new one, affecting the structure, the format and the cover of the Safety Series. It should be noted, however, that subsequent editions of the Regulations and the Schedules of Requirements will be published in the new style. Tabs

  19. Requirements for a systems-based research and development management process in transport infrastructure engineering

    Rust, Frederik Christoffel

    2015-05-01

    Full Text Available The management of research and development (R&D in the transport infrastructure field is complex due to the multidisciplinary nature of the work. The literature shows that linear R&D models that progress from idea through to consumer product are not suitable for the management of such multi-disciplinary projects. This study focuses on determining the key characteristics required for a systems-based approach to the management of R&D projects. The information and data was compiled from literature reviews, interviews, and an e-mail survey with responses from 42 significant international R&D programmes. The findings confirmed the need for a systems-based approach to R&D management. The study formulated twelve principles or tenets for a new, systems-based approach.

  20. Schedules of requirements for the transport of specified types of radioactive material consignments

    These Schedules have been prepared as a companion document to the 1985 Edition of the Transport Regulations (Safety Series No. 6). As presented here, the Schedules also reflect the corrections and changes implemented by the 1986 Supplement to the Regulations for the Safe Transport of Radioactive Material. Direct cross-referencing to the paragraphs, tables and figures in the Regulations is made, and the use of both the International System of Units (SI) and the formerly accepted, traditional units is continued. In many cases, due to rounding off the numbers, the two values for a given parameter for the two units may differ somewhat. To move toward international use of SI units and to avoid inconsistencies, the values for the SI units are controlling in all cases. These Schedules are provided only as an aid to users of the Regulations and the provisions of the Regulations take precedence over the Schedules. These Schedules also serve as a generic model of the Schedules which are part of modal international regulatory documents, such as the International Maritime Dangerous Goods (IMDG) Code of the International Maritime Organization (IMO), the European Agreement Concerning the International Carriage of Dangerous Goods by Road (ADR) and the International Convention Concerning the Carriage of Dangerous Goods by Rail (CIM/RID). For regulatory purposes, reference shall be made to the detailed provisions of the Regulations specified in Safety Series No. 6. The Schedules do not reproduce the provisions of the Regulations in detail nor do they contain any additional requirements. They merely provide a summary of the main provisions for each specified type of radioactive material, references being provided to the relevant detailed provisions of the Regulations to enable these to be consulted where necessary. A table summarizing package, shipment approval, and notification requirements of the Regulations is provided

  1. Piperlongumine selectively suppresses ABC-DLBCL through inhibition of NF-κB p65 subunit nuclear import

    Constitutive NF-κB activation is required for survival of activated B cell-like subtype of diffuse large B cell lymphoma (ABC-DLBCL). However, current NF-κB targeting strategies lack cancer cell specificity. Here, we identified a novel inhibitor, piperlongumine, features direct binding to NF-κB p65 subunit and suppression of p65 nuclear import. This was accompanied by NF-κB reporter activity suppression and NF-κB target gene downregulation. Moreover, mutation of Cys38 to Ser in p65 abolished this effect of piperlongumine on inhibition of p65 nuclear import. Furthermore, we show that piperlongumine selectively inhibited proliferation and induced apoptosis of ABC-DLBCL cells. Most notably, it has been reported that piperlongumine did not affect normal cells even at high doses and was nontoxic to animals. Hence, our current study provides new insight into piperlongumine's mechanism of action and novel approach to ABC-DLBCL target therapy. - Highlights: • Current NF-κB targeting strategies lack cancer cell specificity. • Piperlongumine inhibits NF-κB p65 subunit nuclear import via directly binding to p65. • Piperlongumine selectively inhibits proliferation of ABC-DLBCL cells. • This study provides a novel approach to ABC-DLBCL target therapy

  2. Piperlongumine selectively suppresses ABC-DLBCL through inhibition of NF-κB p65 subunit nuclear import

    Niu, Mingshan [Blood Diseases Institute, Xuzhou Medical College, Xuzhou, Jiangsu (China); Jiangsu Key Laboratory of Bone Marrow Stem Cell, Xuzhou Medical College, Xuzhou, Jiangsu (China); Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Shen, Yangling; Xu, Xiaoyu [Blood Diseases Institute, Xuzhou Medical College, Xuzhou, Jiangsu (China); Jiangsu Key Laboratory of Bone Marrow Stem Cell, Xuzhou Medical College, Xuzhou, Jiangsu (China); Yao, Yao; Fu, Chunling [Blood Diseases Institute, Xuzhou Medical College, Xuzhou, Jiangsu (China); Jiangsu Key Laboratory of Bone Marrow Stem Cell, Xuzhou Medical College, Xuzhou, Jiangsu (China); Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Yan, Zhiling [Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Wu, Qingyun [Blood Diseases Institute, Xuzhou Medical College, Xuzhou, Jiangsu (China); Jiangsu Key Laboratory of Bone Marrow Stem Cell, Xuzhou Medical College, Xuzhou, Jiangsu (China); Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Cao, Jiang; Sang, Wei [Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Zeng, Lingyu [Blood Diseases Institute, Xuzhou Medical College, Xuzhou, Jiangsu (China); Jiangsu Key Laboratory of Bone Marrow Stem Cell, Xuzhou Medical College, Xuzhou, Jiangsu (China); Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Li, Zhenyu [Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu (China); Liu, Xuejiao, E-mail: liuxuejiao0923@126.com [Insititute of Nervous System Diseases, Xuzhou Medical College, Xuzhou, Jiangsu (China); and others

    2015-07-10

    Constitutive NF-κB activation is required for survival of activated B cell-like subtype of diffuse large B cell lymphoma (ABC-DLBCL). However, current NF-κB targeting strategies lack cancer cell specificity. Here, we identified a novel inhibitor, piperlongumine, features direct binding to NF-κB p65 subunit and suppression of p65 nuclear import. This was accompanied by NF-κB reporter activity suppression and NF-κB target gene downregulation. Moreover, mutation of Cys{sup 38} to Ser in p65 abolished this effect of piperlongumine on inhibition of p65 nuclear import. Furthermore, we show that piperlongumine selectively inhibited proliferation and induced apoptosis of ABC-DLBCL cells. Most notably, it has been reported that piperlongumine did not affect normal cells even at high doses and was nontoxic to animals. Hence, our current study provides new insight into piperlongumine's mechanism of action and novel approach to ABC-DLBCL target therapy. - Highlights: • Current NF-κB targeting strategies lack cancer cell specificity. • Piperlongumine inhibits NF-κB p65 subunit nuclear import via directly binding to p65. • Piperlongumine selectively inhibits proliferation of ABC-DLBCL cells. • This study provides a novel approach to ABC-DLBCL target therapy.

  3. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M.; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L.

    2016-01-01

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rh...

  4. Women: the ABC of food security.

    Arcellana, N P

    1997-12-01

    While the 1996 World Food Summit Plan of Action was being approved, a companion NGO (nongovernmental organization) Forum provided opportunities for rural women from 29 countries to relay their perspectives and recommendations. The Rural Women's Workshop was organized by four NGOs: Isis International-Manila, La Via Campesina, the People-Centred Development Forum, and the Women's Food and Agriculture Working Group. Isis International-Manila seeks to create spaces, facilitate processes, and disseminate information for rural women to voice concerns, network, and plan responses. The La Via Campesina network operates in Latin American and the Caribbean where it applies a strong gender perspective to all of its activities. Ultimate progress on the World Food Summit Plan of Action can be evaluated using the ABCs of food security: does the program or policy assure 1) access for women to the total means of production; 2) benefits for women; and 3) community-based resource management and sustainable agriculture. PMID:12321562

  5. WD60/FAP163 is a dynein intermediate chain required for retrograde intraflagellar transport in cilia.

    Patel-King, Ramila S; Gilberti, Renée M; Hom, Erik F Y; King, Stephen M

    2013-09-01

    Retrograde intraflagellar transport (IFT) is required for assembly of cilia. We identify a Chlamydomonas flagellar protein (flagellar-associated protein 163 [FAP163]) as being closely related to the D1bIC(FAP133) intermediate chain (IC) of the dynein that powers this movement. Biochemical analysis revealed that FAP163 is present in the flagellar matrix and is actively trafficked by IFT. Furthermore, FAP163 copurified with D1bIC(FAP133) and the LC8 dynein light chain, indicating that it is an integral component of the retrograde IFT dynein. To assess the functional role of FAP163, we generated an RNA interference knockdown of the orthologous protein (WD60) in planaria. The Smed-wd60(RNAi) animals had a severe ciliary assembly defect that dramatically compromised whole-organism motility. Most cilia were present as short stubs that had accumulated large quantities of IFT particle-like material between the doublet microtubules and the membrane. The few remaining approximately full-length cilia had a chaotic beat with a frequency reduced from 24 to ∼10 Hz. Thus WD60/FAP163 is a dynein IC that is absolutely required for retrograde IFT and ciliary assembly. PMID:23864713

  6. ABC transporter genes and risk of type 2 diabetes

    Schou, Jesper; Tybjærg-Hansen, Anne; Møller, Holger J; Nordestgaard, Børge; Frikke-Schmidt, Ruth

    2012-01-01

    risk of type 2 diabetes in the general population is unknown. RESEARCH DESIGN AND METHODS: We tested whether genetic variation in the promoter and coding regions of ABCA1 and ABCG1 predicted risk of type 2 diabetes in the general population. Twenty-seven variants, identified by previous resequencing of...... of the variants examined, ABCG1 g.-530A>G, predicted a decreased risk of type 2 diabetes in the CCHS (P for trend = 0.05). Furthermore, when validated in the CGPS or in the CCHS and CGPS combined (n = 40,600), neither the two loss-of-function mutations (ABCA1 N1800H, ABCG1 g.-376C>T) nor ABCG1 g.-530......A>G were associated with type 2 diabetes (P values >0.57 and >0.30, respectively). CONCLUSIONS: Genetic variations in ABCA1 and ABCG1 were not associated with increased risk of type 2 diabetes in the general population. These data were obtained in general population samples harboring the largest...

  7. abc: An Extensible AspectJ Compiler

    Avgustinov, Pavel; Christensen, Aske Simon; Hendren, Laurie J.;

    2006-01-01

    checking and code generation, as well as data flow and control flow analyses. The AspectBench Compiler (abc) is an implementation of such a workbench. The base version of abc implements the full AspectJ language. Its front end is built using the Polyglot framework, as a modular extension of the Java...... language. The use of Polyglot gives flexibility of syntax and type checking. The back end is built using the Soot framework, to give modular code generation and analyses. In this paper, we outline the design of abc, focusing mostly on how the design supports extensibility. We then provide a general...

  8. 75 FR 63 - Hazardous Materials: Revision to Requirements for the Transportation of Batteries and Battery...

    2010-01-04

    ...-Powered Devices; and Harmonization With the United Nations Recommendations, International Maritime... of Dangerous Goods Regulations, and the United Nations Recommendations on the Transport of Dangerous... Regulations), and the United Nations Recommendations on the Transport of Dangerous Goods (UN...

  9. Waste transport and storage: Packaging refused due to failure in fulfilling QC/QA requirements

    final disposal of radioactive wastes, Brazilian Competent Authority specifies minimum performance requirements for operations that give wastes a suitable form for storage, transport and disposal. Taking into account the difficulties to demonstrate that the packagings were in compliance with such requirements, the packagings were refused. (author)

  10. Enhanced form of solving real coded numerical optimization problem using ABC algorithm with linear crossover operator

    Mrs. Shanana Gajala Qureshi; Mrs. Uzma Arshi Ansari

    2012-01-01

    Artificial Bee Colony (ABC) algorithm is population-based swarm intelligence algorithm. There are many algorithms Present for solving numeric optimization problem.ABC is based on the intellectual behaviour of honey bee crowd. In this work, ABC algorithm is used with Genetic crossover machinist and tested on standard benchmark functions, and also result compared with the X-ABC algorithm with the advantage of fewer organize Parameters. Obtained results show that the performance of the ABC with ...

  11. Negative membrane curvature catalyzes nucleation of endosomal sorting complex required for transport (ESCRT)-III assembly.

    Lee, Il-Hyung; Kai, Hiroyuki; Carlson, Lars-Anders; Groves, Jay T; Hurley, James H

    2015-12-29

    The endosomal sorting complexes required for transport (ESCRT) machinery functions in HIV-1 budding, cytokinesis, multivesicular body biogenesis, and other pathways, in the course of which it interacts with concave membrane necks and bud rims. To test the role of membrane shape in regulating ESCRT assembly, we nanofabricated templates for invaginated supported lipid bilayers. The assembly of the core ESCRT-III subunit CHMP4B/Snf7 is preferentially nucleated in the resulting 100-nm-deep membrane concavities. ESCRT-II and CHMP6 accelerate CHMP4B assembly by increasing the concentration of nucleation seeds. Superresolution imaging was used to visualize CHMP4B/Snf7 concentration in a negatively curved annulus at the rim of the invagination. Although Snf7 assemblies nucleate slowly on flat membranes, outward growth onto the flat membrane is efficiently nucleated at invaginations. The nucleation behavior provides a biophysical explanation for the timing of ESCRT-III recruitment and membrane scission in HIV-1 budding. PMID:26668364

  12. Redundancy Level Optimization in Modular Software System Models using ABC

    Tarun Kumar Sharma

    2014-03-01

    Full Text Available The performance of optimization algorithms is problem dependent and as per no free lunch theorem, there exists no such algorithm which can be efficiently applied to every type of problem(s. However, we can modify the algorithm/ technique in a manner such that it is able to deal with a maximum type of problems. In this study we have modified the structure of basic Artificial Bee Colony (ABC, a recently proposed metaheuristic algorithm based on the concept of swarm intelligence to optimize the models of software reliability. The modified variant of ABC is termed as balanced ABC (B-ABC. The simulated results show the efficiency and capability of the variant to solve such type of the problems.

  13. Yang-Mills Theory and the ABC Conjecture

    He, Yang-Hui; Probst, Malte; Read, James

    2016-01-01

    We establish a precise correspondence between the ABC Conjecture and N=4 super-Yang-Mills theory. This is achieved by combining three ingredients: (i) Elkies' method of mapping ABC-triples to elliptic curves in his demonstration that ABC implies Mordell/Faltings; (ii) an explicit pair of elliptic curve and associated Belyi map given by Khadjavi-Scharaschkin; and (iii) the fact that the bipartite brane-tiling/dimer model for a gauge theory with toric moduli space is a particular dessin d'enfant in the sense of Grothendieck. We explore this correspondence for the highest quality ABC-triples as well as large samples of random triples. The Conjecture itself is mapped to a statement about the fundamental domain of the toroidal compactification of the string realization of N=4 SYM.

  14. Multicompartment Micelles From π-Shaped ABC Block Copolymers

    XIA Jun; ZHONG Chong-Li

    2007-01-01

    Dissipative particle dynamics simulations were performed on the morphology and structure of multicompartment micelles formed from n-shaped ABC block copolymers in water. The influences of chain architectures were studied in a systematic way, and a rich variety of morphologies were observed, such as spherical, wormlike,X-shaped, Y-shaped, ribbon-like, layered rod-like, layered disk-like, as well as network morphologies. The simulations show that the distance between the two grafts plays an important role in control of the morphology. Since π-shaped ABC block copolymers can be reduced to linear ABC and star ABC block copolymers, they are good model copolymers for studying the self-assembly of complex block copolymers into micelles. The knowledge obtained in this work as well as the new morphologies identified provide useful information for future rational design and synthesis of novel multicompartment micelles.

  15. The saci_2123 gene of the hyperthermoacidophile Sulfolobus acidocaldarius encodes an ATP-binding cassette multidrug transporter

    Yang, Nuan; Driessen, Arnold J. M.

    2015-01-01

    Multidrug resistance (MDR) transporters are capable of secreting structurally and functionally unrelated toxic compounds from the cell. Among this group are ATP-binding cassette (ABC) transporters. These membrane proteins are typically arranged as either hetero- or homo-dimers of ABC half-transporte

  16. The Saccharomyces cerevisiae v-SNARE Vti1p Is Required for Multiple Membrane Transport Pathways to the Vacuole

    von Mollard, Gabriele Fischer; Stevens, Tom H.

    1999-01-01

    The interaction between v-SNAREs on transport vesicles and t-SNAREs on target membranes is required for membrane traffic in eukaryotic cells. Here we identify Vti1p as the first v-SNARE protein found to be required for biosynthetic traffic into the yeast vacuole, the equivalent of the mammalian lysosome. Certain vti1-ts yeast mutants are defective in alkaline phosphatase transport from the Golgi to the vacuole and in targeting of aminopeptidase I from the cytosol to the vacuole. VTI1 interact...

  17. Product development in company ABC: Strategic analysis and recommendations

    Tadaniewicz, Darius

    2005-01-01

    ABC is a mature company, operating in the power electronics market, using the stage gate process to develop products. This analysis evaluates the company's development process including innovation and portfolio management. An examination of the market reveals that speed-to-market and commitment to innovation are critical success factors. Benchmarking shows how comparable companies have adapted the traditional stage gate process to incorporate more flexible practices. Analysis of ABC uncovers ...

  18. Product Profitability Analysis Based on EVA and ABC

    Chen Lin; Shuangyuan Wang; Zhilin Qiao

    2013-01-01

    On the purpose of maximizing shareholders’ value, the profitability analysis established on the basis oftraditional accounting earnings cannot meet the demands of providing accurate decision-making information forenterprises. Therefore, this paper implements the Activity Based Costing (ABC) and the Economic Value Added(EVA) into the traditional profitability analysis system, sets up an improved EVA-ABC based profitabilityanalysis system as well as its relative indexes, and applies it to the s...

  19. The 'ABC' of examining foot radiographs.

    Pearse, Eyiyemi O.; Klass, Benjamin; Bendall, Stephen P.

    2005-01-01

    INTRODUCTION: We report a simple systematic method of assessing foot radiographs that improves diagnostic accuracy and can reduce the incidence of inappropriate management of serious forefoot and midfoot injuries, particularly the Lisfranc-type injury. STUDY GROUP AND METHODS: Five recently appointed senior house officers (SHOs), with no casualty or Orthopaedic experience prior to their appointment, were shown a set of 10 foot radiographs and told the history and examination findings recorded in the casualty notes of each patient within 6 weeks of taking up their posts. They were informed that the radiographs might or might not demonstrate an abnormality. They were asked to make a diagnosis and decide on a management plan. The test was repeated after they were taught the 'ABC' method of evaluating foot radiographs. RESULTS: Diagnostic accuracy improved after SHOs were taught a systematic method of assessing foot radiographs. The proportion of correct diagnoses increased from 0.64 to 0.78 and the probability of recognising Lisfranc injuries increased from 0 to 0.6. CONCLUSIONS: The use of this simple method of assessing foot radiographs can reduce the incidence of inappropriate management of serious foot injuries by casualty SHOs, in particular the Lisfranc type injury. PMID:16263015

  20. Double pionic fusion. Towards an understanding of the ABC puzzle by exclusive measurements

    The ABC effect is a huge unexpected enhancement at twice the pion mass in the invariant mass spectrum of two pions, which are generated in double-pionic fusion to bound nuclear systems. This peculiar phenomenon has been missing a conclusive explanation all the time since it has been discovered 1960 in single-arm measurements of 3He ejectiles in the reaction pd→ 3HeX. One reason for this failure has been that all measurements to this subject have been inclusive, i.e., lacking the full experimentally accessible information. Hence exclusive measurements were performed at CELSIUS/WASA at an energy of Tp=0.895 GeV, where the ABC effect is expected to be strongest. For the first time exclusive data of solid statistics for both the pd→3Heπ0π0 and pd→3Heπ+π- reactions were obtained including also results for the three-pion production total cross-section. The new data are consistent with the previous inclusive data. They provide, however, much more additional information, which rule out all previous explications of the ABC effect. The now available kinematically complete set of data reveals that the low ππ-mass enhancement (ABC-effect): - is not necessarily associated with a high ππ-mass enhancement, - is always connected with the simultaneous excitation of two Δ resonances, - is of scalar-isoscalar nature, i.e. a σ-channel phenomenon, - requires dynamics in the reaction system, which has not been considered hitherto. Various possible solutions are discussed, however, all of them demand a high attraction in the ΔΔ system - a point, which has never been touched so far in theoretical and experimental investigations. For this data analysis new powerful methods based on neural nets have been developed. Their current and possible future applications are discussed. (orig.)

  1. A single-amino-acid substitution eliminates the stringent carbohydrate requirement for intracellular transport of a viral glycoprotein.

    Pitta, A M; Rose, J. K.; Machamer, C.E.

    1989-01-01

    In this report, we have investigated the contribution of primary sequence to the carbohydrate requirement for intracellular transport of two closely related glycoproteins, the G proteins of the San Juan and Orsay strains of vesicular stomatitis virus. We used site-directed mutagenesis of the coding sequence to eliminate the two consensus sites for glycosylation in the Orsay G protein. Whereas the nonglycosylated San Juan G protein required at least one of its two asparagine-linked oligosaccha...

  2. The N terminus of monoamine transporters is a lever required for the action of amphetamines

    Sucic, Sonja; Dallinger, Stefan; Zdrazil, Barbara;

    2010-01-01

    The serotonin transporter (SERT) terminates neurotransmission by removing serotonin from the synaptic cleft. In addition, it is the site of action of antidepressants (which block the transporter) and of amphetamines (which induce substrate efflux). We explored the functional importance of the N t...

  3. 9 CFR 11.40 - Prohibitions and requirements concerning persons involved in transportation of certain horses.

    2010-01-01

    ... concerning persons involved in transportation of certain horses. 11.40 Section 11.40 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE ANIMAL WELFARE HORSE... certain horses. (a) Each person who ships, transports, or otherwise moves, or delivers or receives...

  4. Bacterial multidrug resistance mediated by a homologue of the human multidrug transporter P-glycoprotein

    Konings, WN; Poelarends, GJ

    2002-01-01

    Most ATP-binding cassette (ABC) multidrug transporters known to date are of eukaryotic origin, such as the P-glycoproteins (Pgps) and multidrug resistance-associated proteins (MRPs). Only one well-characterized ABC multidrug transporter, LmrA, is of bacterial origin. On the basis of its structural a

  5. 40 CFR 51.916 - What are the requirements for an Ozone Transport Region under the 8-hour NAAQS?

    2010-07-01

    ... 40 Protection of Environment 2 2010-07-01 2010-07-01 false What are the requirements for an Ozone Transport Region under the 8-hour NAAQS? 51.916 Section 51.916 Protection of Environment ENVIRONMENTAL... IMPLEMENTATION PLANS Provisions for Implementation of 8-hour Ozone National Ambient Air Quality Standard §...

  6. Metodologia ABC: implantação numa microempresa ABC methodology: implementation on a micro firm

    Orlando Duran

    2000-08-01

    Full Text Available Este trabalho relata a implantação da metodologia de custos baseados em atividades numa microempresa do ramo metalúrgico. A proposta pretende demonstrar a viabilidade de aplicar esta técnica em empresas sem importar seu tamanho, só realizando algumas adaptações que garantam baixo investimento e curto espaço de tempo para obter os resultados. Na parte final do trabalho se realiza uma análise dos resultados obtidos verificando-se o potencial da informação gerada pela metodologia e seu uso como ferramenta de gestão.This paper presents an implementation of the activity based costing (ABC methodology in a small firm. The approach presented is intended to demonstrate the feasibility of applying the ABC methodology at any sized firm, only through few adaptations for ensuring low investments fees and speed in obtaining results and information from the system. Discussion about the results obtained during the implementation case are presented and the potential of using the information generated from the system as a managing tool is commented.

  7. Transport

    Transport is one of the major causes of environmental damage in Austria. Energy consumption, pollutants emissions, noise emissions, use of surfaces, sealing of surfaces, dissection of ecosystems and impact on landscape are the most significant environmental impacts caused by it. An overview of the transport development of passengers and freight in Austria is presented. Especially the energy consumption growth, carbon dioxide and nitrogen oxide emissions by type of transport, and the emissions development (HC, particle and carbon monoxide) of goods and passengers transport are analyzed covering the years 1980 - 1999. The health cost resulting from transport-related air pollution in Austria is given and measures to be taken for an effective control of the transport sector are mentioned. Figs. 8, Table 1. (nevyjel)

  8. TTC26/DYF13 is an intraflagellar transport protein required for transport of motility-related proteins into flagella

    Ishikawa, Hiroaki; Ide, Takahiro; Yagi, Toshiki; Jiang, Xue; Hirono, Masafumi; Sasaki, Hiroyuki; Yanagisawa, Haruaki; Wemmer, Kimberly A; Stainier, Didier YR; Qin, Hongmin; Kamiya, Ritsu; Marshall, Wallace F

    2014-01-01

    Cilia/flagella are assembled and maintained by the process of intraflagellar transport (IFT), a highly conserved mechanism involving more than 20 IFT proteins. However, the functions of individual IFT proteins are mostly unclear. To help address this issue, we focused on a putative IFT protein TTC26/DYF13. Using live imaging and biochemical approaches we show that TTC26/DYF13 is an IFT complex B protein in mammalian cells and Chlamydomonas reinhardtii. Knockdown of TTC26/DYF13 in zebrafish embryos or mutation of TTC26/DYF13 in C. reinhardtii, produced short cilia with abnormal motility. Surprisingly, IFT particle assembly and speed were normal in dyf13 mutant flagella, unlike in other IFT complex B mutants. Proteomic and biochemical analyses indicated a particular set of proteins involved in motility was specifically depleted in the dyf13 mutant. These results support the concept that different IFT proteins are responsible for different cargo subsets, providing a possible explanation for the complexity of the IFT machinery. DOI: http://dx.doi.org/10.7554/eLife.01566.001 PMID:24596149

  9. CFTR is required for maximal transepithelial liquid transport in pig alveolar epithelia

    Li, Xiaopeng; Comellas, Alejandro P.; Karp, Philip H.; Ernst, Sarah E.; Moninger, Thomas O.; Gansemer, Nicholas D.; Taft, Peter J.; Pezzulo, Alejandro A; Michael V Rector; Rossen, Nathan; Stoltz, David A.; McCray, Paul B.; Welsh, Michael J.; Zabner, Joseph

    2012-01-01

    A balance between alveolar liquid absorption and secretion is critical for maintaining optimal alveolar subphase liquid height and facilitating gas exchange in the alveolar space. However, the role of cystic fibrosis transmembrane regulator protein (CFTR) in this homeostatic process has remained elusive. Using a newly developed porcine model of cystic fibrosis, in which CFTR is absent, we investigated ion transport properties and alveolar liquid transport in isolated type II alveolar epitheli...

  10. An Arabidopsis flavonoid transporter is required for anther dehiscence and pollen development

    Thompson, Elinor P.; Wilkins, Christopher; Demidchik, Vadim; Davies, Julia M; Glover, Beverley J.

    2010-01-01

    FLOWER FLAVONOID TRANSPORTER (FFT) encodes a multidrug and toxin efflux family transporter in Arabidopsis thaliana. FFT (AtDTX35) is highly transcribed in floral tissues, the transcript being localized to epidermal guard cells, including those of the anthers, stigma, siliques and nectaries. Mutant analysis demonstrates that the absence of FFT transcript affects flavonoid levels in the plant and that the altered flavonoid metabolism has wide-ranging consequences. Root growth, seed development ...

  11. Astrocyte-neuron lactate transport is required for long-term memory formation

    Suzuki, Akinobu; Stern, Sarah A.; Bozdagi, Ozlem; Huntley, George W.; Walker, Ruth H.; Magistretti, Pierre J.; Alberini, Cristina M

    2011-01-01

    We report that in the rat hippocampus learning leads to a significant increase in extracellular lactate levels, which derive from glycogen, an energy reserve selectively localized in astrocytes. Astrocytic glycogen breakdown and lactate release are essential for long-term but not short-term memory formation, and for the maintenance of long-term potentiation (LTP) of synaptic strength elicited in-vivo. Disrupting the expression of the astrocytic lactate transporters monocarboxylate transporter...

  12. Golgi-localized UDP-glucose transporter is required for cell wall integrity in rice

    Song, Xueqin; Zhang, Baocai; Zhou, Yihua

    2011-01-01

    Cell wall-related nucleotide sugar transporters (NSTs) theoretically supply the cytosolic nucleotide sugars for glycosyltransferases (GTs) to carry out ploysaccharide synthesis and modification in the Golgi apparatus. However, the regulation of cell wall synthesis by NSTs remains undescribed. Recently, we have reported the functional characterization of Oryza sativa nucleotide sugar transport (Osnst1) mutant and its corresponding gene. OsNST1/BC14 is localized in the Golgi apparatus and trans...

  13. General requirement for harvesting antennae at Ca2+ and H+ channels and transporters

    Cristián Martínez

    2010-09-01

    Full Text Available The production and dissipation of energy in cells is intimately linked to the movement of small molecules in and out of enzymes, channels and transporters. An analytical model of diffusion was described previously, which was used to estimate local effects of these proteins acting as molecular sources. The present article describes a simple but more general model, which can be used to estimate the local impact of proteins acting as molecular sinks. The results show that the enzymes, transporters and channels, whose substrates are present at relatively high concentrations like ATP, Na+, glucose, lactate and pyruvate, do not operate fast enough to deplete their vicinity to a meaningful extent, supporting the notion that for these molecules the cytosol is a well-mixed compartment. One specific consequence of this analysis is that the well-documented cross-talk existing between the Na+/K+ ATPase and the glycolytic machinery should not be explained by putative changes in local ATP concentration. In contrast, Ca2+ and H+ transporters like the Na+/Ca2+ exchanger NCX and the Na+/H+ exchanger NHE, show experimental rates of transport that are two to three orders of magnitude faster than the rates at which the aqueous phase may possibly feed their binding sites. This paradoxical result implies that Ca2+ and H+ transporters do not extract their substrates directly from the bulk cytosol, but from an intermediate “harvesting” compartment located between the aqueous phase and the transport site.

  14. Transportation of radioactive materials: a summary of state and local legislative requirements for the period ending December 31, 1985

    This report lists 670 adopted US state and local laws that impact the transportation of radioactive materials. The report was generated from information contained in the Legislative Database (LDB), a comprehensive, interactive database developed at the Oak Ridge National Laboratory for the US Department of Energy and the Joint Integration Office. Laws are sorted alphabetically by state, with state and local bills listed separately and sorted by date of adoption. Each citation contains the following information: locale (geographical areas and political jurisdictions affected by the action), bill number, bill title, bill sponsor, history of bill status, comments, and abstract. Seven indexes are provided to assist the reader in locating legislation of interest: locale, bill number, title word (permuted), sponsor, transport restriction (type of transportation restriction specified, e.g., escort, notify, permit, ban), transport mode (mode of transportation specified, e.g., truck, rail, barge), and keyword. This report adds new legislation to the information contained in last year's report, ''Transportation of Radioactive and Hazardous Materials: A Summary of State and Local Legislative Requirements for the Period Ending December 31, 1981,'' ORNL/TM-9563, published in September 1985

  15. Development of an improved radiological basis and revised requirements for the transport of LSA/SCO materials

    The IAEA Regulations for the Safe Transport of Radioactive Material specify a range of package requirements that are dependent upon the activity and properties of the radioactive material being transported. For example, Type A packages can be used for the transport of material with an activity of up to one A1 of special form material and one A2 of other material. The A1 and A2 values are determined for over 350 radionuclides using the Q system in the advisory material. This system models the following pathways in the event of a severe accident: External photon dose; External beta dose; Inhalation dose; Skin and ingestion dose due to contamination transfer; Submersion dose. Industrial packages can be used for the transport of low specific activity (LSA) material and surface contaminated objects (SCO) and the particular requirements for these types of materials are derived from a simplistic radiological model that is often referred to as the 10 mg inhalation model. This is because it is based on the assumption that, in a dusty environment, an individual would inhale no more than 10 mg in such a situation, although there is limited reliable information to justify this assumption. Furthermore, there are deficiencies in the specification of some of the material requirements for LSA/SCO material in the IAEA Transport Regulations. Consequently, this can cause some difficulties for operators in complying with the regulations and to competent authorities in their compliance assurance role. Particular areas of difficulty include: It can be difficult to distinguish between LSA-type material and SCO-type; It is difficult to distinguish surface contamination from activity within the object, and between fixed and nonfixed contamination; In order to demonstrate compliance with SCO requirements, measurements of both accessible and nonaccessible contamination are required, although it is not clear how compliance with the inaccessible contamination limit can be measured if it cannot

  16. Enhanced form of solving real coded numerical optimization problem using ABC algorithm with linear crossover operator

    Mrs. Shanana Gajala Qureshi

    2012-06-01

    Full Text Available Artificial Bee Colony (ABC algorithm is population-based swarm intelligence algorithm. There are many algorithms Present for solving numeric optimization problem.ABC is based on the intellectual behaviour of honey bee crowd. In this work, ABC algorithm is used with Genetic crossover machinist and tested on standard benchmark functions, and also result compared with the X-ABC algorithm with the advantage of fewer organize Parameters. Obtained results show that the performance of the ABC with crossover gives better results than the X-ABC algorithm.

  17. Cuban experience in verification of the execution of the safety requirements during the transport of radioactive materials

    The Cuban Regulatory Authority has paid special attention to the verification of the execution of the safety requirements during the transport of radioactive material in the country. With this purpose, the Authority has followed a consequent policy based on supplementary demands to those collections in the juridical mark settled down in 1987 in the sphere of transport of radioactive substances. In the work the technical approaches are exposed kept in mind when establishing the one referred politics, the current situation is characterized, the results are evaluated obtained in correspondence with the pursued objectives and the essential aspects are exposed to keep in mind for the adopted politics ulterior development. (author)

  18. Classification of the railway accident in accordance with the requirement of the safety analysis of transporting spent fuel

    Based on the analysis of the difference between the accident severity categorization used in the Ministry of Railway and that used in the safety analysis of the transporting spent fuel, a method used for the classification of the railway accident in accordance with the requirement of the safety analysis of transporting spent fuel is suggested. The method classifies the railway accidents into 10 scenarios and make it possible to scale the accident through directly using the data documented by the Ministry of Railway without any additional effort

  19. 军用运输机集装运输托盘需求分析%Analysis of General Requirement of Military Transport Airplane Container Transport Pallets

    张家应; 刘子明

    2015-01-01

    In order to provide some theoretical references for the development of military transport airplane container transport pallet, the requirements on every aspect of the development military transport airplane container transport pallet, including its operational functions,support environment,suitable model,tactical and technical capabilities performance indexes, specifications,were analyzed and discussed in this paper.%对军用运输机集装运输托盘研制的业务功能、保障环境、适用机型和技术规范标准等方面的需求进行初步分析,提出了军用运输机集装运输托盘研制需求的详细内容。随着托盘设计研究的不断深入,有关需求的内容与标准要求还应随之不断补充调整。

  20. Critical evaluation of safety and radiological protection requirements adopted for the transport of uranium and thorium ores and concentrates

    This work evaluates in a critical way the safety and radiological protection recommendations established by the International Atomic Energy Agency - IAEA and adopted national and internationally, for the transport of uranium and thorium ores and concentrates, known according the transport regulations, as being of the Low Specific Activity Material Type-I, LSA-I, basing on more realistic scenarios than the presently existent, aiming at the determination of maximum exposure levels of radiation as well as the maximal contents of those materials in packages and conveyance. A general overview taking into account the scenarios foreseen by the regulations of the IAEA pointed out for a need of a better justification of the requirements edited by the Agency or should be used to support a request of revision of those regulations, national and internationally adopted, in the pertinent aspects to the transport of uranium and thorium ores and concentrates. (author)

  1. Second-order nonlinear optical metamaterials: ABC-type nanolaminates

    We demonstrate a concept for second-order nonlinear metamaterials that can be obtained from non-metallic centrosymmetric constituents with inherently low optical absorption. The concept is based on iterative atomic-layer deposition of three different materials, A = Al2O3, B = TiO2, and C = HfO2. The centrosymmetry of the resulting ABC stack is broken since the ABC and the inverted CBA sequences are not equivalent—a necessary condition for non-zero second-order nonlinearity. In our experiments, we find that the bulk second-order nonlinear susceptibility depends on the density of interfaces, leading to a nonlinear susceptibility of 0.26 pm/V at a wavelength of 800 nm. ABC-type nanolaminates can be deposited on virtually any substrate and offer a promising route towards engineering of second-order optical nonlinearities at both infrared and visible wavelengths

  2. Spent fuel storage and transport cask decontamination and modification. An overview of management requirements and applications based on practical experience

    A large increase in the number of casks required for transport and/or storage of spent fuel is forecast into the next century. The principal requirement will be for increased number of storage and dual purpose (transport/storage) casks for interim storage of spent fuel prior to reprocessing or permanent disposal in both on-site and off-site storage facilities. Through contact with radioactive materials spent fuel casks will be contaminated on both internal and external surfaces. In broad terms, cask contamination management can be defined by three components: minimisation, prevention and decontamination. This publication is a compilation of international experience with cask contamination problems and decontamination practices. The objective is to present current knowledge and experience as well as developments, trends and potential for new applications in this field. Furthermore, the report may assist in new design or modification of existing casks, cask handling systems and decontamination equipment

  3. Requirement of kinesin-mediated membrane transport of WAVE2 along microtubules for lamellipodia formation promoted by hepatocyte growth factor.

    Takahashi, Kazuhide; Suzuki, Katsuo

    2008-07-01

    Lamellipodia formation necessary for epithelial cell migration and invasion is accomplished by rearrangement of the actin cytoskeleton at the leading edge through membrane transport of WAVE2. However, how WAVE2 is transported to the cell periphery where lamellipodia are formed remains to be established. We report here that hepatocyte growth factor (HGF) promoted lamellipodia formation and intracellular transport of WAVE2 to the cell periphery, depending on Rac1 activity, in MDA-MB-231 human breast cancer cells. Immunoblot analyses indicating the coimmunoprecipitation of WAVE2 with kinesin heavy chain KIF5B, one of the motor proteins, and IQGAP1 suggest that KIF5B and IQGAP1 formed a complex with WAVE2 in serum-starved cells and increased in their amount after HGF stimulation. Both downregulation of KIF5B by the small interfering RNA and depolymerization of microtubules with nocodazole abrogated the HGF-induced lamellipodia formation and WAVE2 transport. Therefore, we propose here that the promotion of lamellipodia formation by HGF in MDA-MB-231 cells is Rac1-dependent and requires KIF5B-mediated transport of WAVE2 and IQGAP1 to the cell periphery along microtubules. PMID:18514191

  4. 77 FR 17394 - Hazardous Materials: Approval and Communication Requirements for the Safe Transportation of Air...

    2012-03-26

    ... Department of Transportation (44 FR 11034). Executive Order 13563 is supplemental to and reaffirms the... tape or a waterproof soft plastic cap that must easily provide ventilation of the drum contents in the... has substantial direct effects on the States, the relationship between the national government and...

  5. Repellent Taxis in Response to Nickel Ion Requires neither Ni2+ Transport nor the Periplasmic NikA Binding Protein▿

    Englert, Derek L.; Adase, Christopher A.; Jayaraman, Arul; Manson, Michael D.

    2010-01-01

    Ni2+ and Co2+ are sensed as repellents by the Escherichia coli Tar chemoreceptor. The periplasmic Ni2+ binding protein, NikA, has been suggested to sense Ni2+. We show here that neither NikA nor the membrane-bound NikB and NikC proteins of the Ni2+ transport system are required for repellent taxis in response to Ni2+.

  6. Review of data requirements for groundwater flow and solute transport modelling and the ability of site investigation methods to meet these requirements

    This report describes the data requirements for the codes that may be used in the modelling of groundwater flow and radionuclide transport during the assessment of a Nirex site for the deep disposal of low and intermediate level radioactive waste and also the site investigation methods that exist to supply the data for these codes. The data requirements for eight codes are reviewed, with most emphasis on three of the more significant codes, VANDAL, NAMMU and CHEMTARD. The largest part of the report describes and discusses the site investigation techniques and each technique is considered in terms of its ability to provide the data necessary to characterise the geological and hydrogeological environment around a potential repository. (author)

  7. The membrane QmoABC complex interacts directly with the dissimilatory adenosine 5´-phosphosulfate reductase in sulfate reducing bacteria

    Ana Raquel Ramos

    2012-04-01

    Full Text Available The adenosine 5’-phosphosulfate reductase (AprAB is the enzyme responsible for the reduction of adenosine 5’-phosphosulfate (APS to sulfite in the biological process of dissimilatory sulfate reduction, which is carried out by a ubiquitous group of sulfate reducing prokaryotes. The electron donor for AprAB has not been clearly identified, but was proposed to be the QmoABC membrane complex, since an aprBA-qmoABC gene cluster is found in many sulfate-reducing and sulfur-oxidising bacteria. The QmoABC complex is essential for sulfate reduction, but electron transfer between QmoABC and AprAB has not been reported. In this work we provide the first direct evidence that QmoABC and AprAB interact in Desulfovibrio spp., using co-immunoprecipitation, cross-linking Far-Western blot, tag-affinity purification and surface plasmon resonance studies. This showed that the QmoABC-AprAB complex has a strong steady-state affinity (KD = 90 ± 3 nM, but has a transient character due to a fast dissociation rate. Far-Western blot identified QmoA as the Qmo subunit most involved in the interaction. Nevertheless, electron transfer from menaquinol analogues to APS through anaerobically purified QmoABC and AprAB could not be detected. We propose that this reaction requires the involvement of a third partner to allow electron flow driven by a reverse electron bifurcation process i.e. electron confurcation. This process is deemed essential to allow coupling of APS reduction to chemiosmotic energy conservation.

  8. Tumor metastatic promoter ABCE1 interacts with the cytoskeleton protein actin and increases cell motility.

    Han, Xu; Tian, Ye; Tian, Dali

    2016-06-01

    ABCE1, a member of the ATP-binding cassette (ABC) family, is a candidate tumor metastatic promoter in lung cancer. Overexpression of ABCE1 is correlated with aggressive growth and metastasis in lung cancer cells. However, the exact mechanism remains unclear. In the present study, GST pull-down assay provided evidence of the possible interaction between ABCE1 and β-actin using GST-ABCE1 as a bait protein. Co-immunoprecipitation manifested ABCE1 formed complexes with β-actin in vivo. ABCE1 overexpression significantly increased the migration of lung cancer cells which may be attributed to the promotion of F-actin rearrangements. Taken together, these data suggest that overexpression of ABCE1 produces an obvious effect on the motility of lung cancer cells through cytoskeleton rearrangement. PMID:27109616

  9. Glutathione-Conjugate Transport by RLIP76 is required for Clathrin-Dependent Endocytosis and Chemical Carcinogenesis**

    SINGHAL, SHARAD S.; Wickramarachchi, Dilki; Yadav, Sushma; Singhal, Jyotsana; Leake, Kathryn; Vatsyayan, Rit; Chaudhary, Pankaj; Lelsani, Poorna; Suzuki, Sumihiro; Yang, Shaohua; Awasthi, Yogesh C.; Awasthi, Sanjay

    2011-01-01

    Targeted depletion of the RALBP1 encoded 76 kDa splice variant, RLIP76, causes marked and sustained regression of human xenografts of lung, colon, prostate, and kidney cancer without toxicity in nude mouse models. We proposed that the remarkable efficacy and broad-spectrum of RLIP76-targeted therapy is because its glutathione-conjugate (GS-E) transport-activity is required for clathrin-dependent endocytosis (CDE), that regulates all ligand-receptor signaling, and that RLIP76 is required not o...

  10. Screening of miRNA regulating ABCE1 gene in non-small-cell lung cancer%ABCE1基因在非小细胞肺癌内相关调节miRNA的筛选

    田野; 刘思洋; 许辉; 姜文军; 赵希彤; 王晴; 田大力

    2015-01-01

    Objective To screen the miRNAs regulating ATP-binding cassette transporter E1(ABCE1) gene in non-small-cell lung cancer, and explore new strategies in lung cancer diagnosis and therapy. Methods The 20 patients with non-small-cell lung cancer(11 squamous cell carcinoma and 9 adenocarcinoma) were enrolled, included 13 males and 7 females, which aged 45-73 years old with mean age of 62.9 years old. Bioinformatics was used to predict the miRNAs regulated ABCE1 gene;statistical analysis was then done to screen out the purpose miRNA by real-time quantitative PCR(RT-Q-PCR) and detected miRNAs and ABCE1 mRNA and protein. Results The result of bioinformatics software predicted that seven miRNAs had highest possibility to regulate ABCE1 gene, which were miR-29a/b/c, miR-135a/b, miR-203 and miR-141. The expression of miR-29a/b/c, miR-135a, miR-203, especially miR-135a and miR-29c in carcinoma tissues, compared to those in pericarcinomatous tissues experienced decrease to different degrees, while the expression of mRNA and protein of ABCE1 increased in carcinoma tissues ( P < 0.05). Moreover, there appeared to be negative correlation between miR-135a and ABCE1 in lung cancer tissues(r=-0.665,P=0.001). Conclusion It is demonstrated that the miR-135a negatively regulates ABCE1 gene, and the combination of them might be the new target for diagnosis and treatment of non-small-cell lung cancer.%目的:筛选ATP结合盒E1(ABCE1)基因的相关调节miRNA,为诊治肺癌提供新思路。方法选取20例非小细胞肺癌患者,其中男性13例,女性7例;年龄45~73岁,平均年龄62.9岁。鳞癌11例,腺癌9例。应用生物信息学预测ABCE1基因上游的miRNA,通过实时定量聚合酶链反应(RT-Q-PCR)及免疫组织化学方法,对标本非小细胞癌组织和癌旁组织进行检测,并进行统计学分析,从中筛选出目的miRNA。结果生物信息软件预测7个最有可能调节 ABCE1基因的miRNA,分别为miR-29a

  11. ABCE1基因在非小细胞肺癌内相关调节miRNA的筛选%Screening of miRNA regulating ABCE1 gene in non-small-cell lung cancer

    田野; 刘思洋; 许辉; 姜文军; 赵希彤; 王晴; 田大力

    2015-01-01

    目的:筛选ATP结合盒E1(ABCE1)基因的相关调节miRNA,为诊治肺癌提供新思路。方法选取20例非小细胞肺癌患者,其中男性13例,女性7例;年龄45~73岁,平均年龄62.9岁。鳞癌11例,腺癌9例。应用生物信息学预测ABCE1基因上游的miRNA,通过实时定量聚合酶链反应(RT-Q-PCR)及免疫组织化学方法,对标本非小细胞癌组织和癌旁组织进行检测,并进行统计学分析,从中筛选出目的miRNA。结果生物信息软件预测7个最有可能调节 ABCE1基因的miRNA,分别为miR-29a/b/c、miR-135a/b、miR-203及miR-141;其中miR-29a/b/c、miR-135a、miR-203的表达在癌组织内较癌旁组织都有不同程度的降低,以miR-135a、miR-29c差异最为明显,与之对应ABCE1在相同的肺癌组织内表达上调(P<0.05);仅miR-135a与ABCE1在上述肺癌患者内表达呈现负性相关(r=-0.665,P=0.001)。结论在非小细胞肺癌内,很有可能是miR-135a负性调节ABCE1基因,两者结合可能成为诊治肺癌的新靶点。%Objective To screen the miRNAs regulating ATP-binding cassette transporter E1(ABCE1) gene in non-small-cell lung cancer, and explore new strategies in lung cancer diagnosis and therapy. Methods The 20 patients with non-small-cell lung cancer(11 squamous cell carcinoma and 9 adenocarcinoma) were enrolled, included 13 males and 7 females, which aged 45-73 years old with mean age of 62.9 years old. Bioinformatics was used to predict the miRNAs regulated ABCE1 gene;statistical analysis was then done to screen out the purpose miRNA by real-time quantitative PCR(RT-Q-PCR) and detected miRNAs and ABCE1 mRNA and protein. Results The result of bioinformatics software predicted that seven miRNAs had highest possibility to regulate ABCE1 gene, which were miR-29a/b/c, miR-135a/b, miR-203 and miR-141. The expression of miR-29a/b/c, miR-135a, miR-203, especially miR-135a and miR-29c in carcinoma tissues, compared to those

  12. Migration of Adipose-derived Mesenchymal Stem Cells Stably Expressing Chondroitinase ABC In vitro

    Jian-Huang Wu; Miao Li; Yan Liang; Tao Lu; Chun-Yue Duan

    2016-01-01

    Background:Several studies have revealed that adipose-derived mesenchymal stem cells (ADSCs) can be used as seed cells for the treatment of spinal cord injury (SCI).Chondroitinase ABC (ChABC) decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI,allowing stem cells to penetrate through the scar and promote recovery of nerve function.This study aimed to establish ADSCs that stably express ChABC (ChABC-ADSCs) and evaluate the migratory capability of ChABC-ADSCs in vitro.Methods:ADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion.Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation.After successful culture,ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained.Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method,ChABC expression was verified using Western blotting,and the migration of ChABC-ADSCs was analyzed using the transwell assay.Results:Secondary collagenase digestion increased the isolation efficiency of primary ADSCs.Following transfection using lentiviral vectors,the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h (P < 0.05).And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group (P < 0.05).Moreover,ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control (P < 0.05).Conclusions:Secondary collagenase digestion can be used to effectively isolate ADSCs.ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC,and ChABC expression significantly enhances the migratory capacity of ADSCs.

  13. Migration of Adipose-derived Mesenchymal Stem Cells Stably Expressing Chondroitinase ABC In vitro

    Wu, Jian-Huang; Li, Miao; Liang, Yan; Lu, Tao; Duan, Chun-Yue

    2016-01-01

    Background: Several studies have revealed that adipose-derived mesenchymal stem cells (ADSCs) can be used as seed cells for the treatment of spinal cord injury (SCI). Chondroitinase ABC (ChABC) decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI, allowing stem cells to penetrate through the scar and promote recovery of nerve function. This study aimed to establish ADSCs that stably express ChABC (ChABC-ADSCs) and evaluate the migratory capability of ChABC-ADSCs in vitro. Methods: ADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion. Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation. After successful culture, ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained. Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method, ChABC expression was verified using Western blotting, and the migration of ChABC-ADSCs was analyzed using the transwell assay. Results: Secondary collagenase digestion increased the isolation efficiency of primary ADSCs. Following transfection using lentiviral vectors, the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h (P < 0.05). And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group (P < 0.05). Moreover, ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control (P < 0.05). Conclusions: Secondary collagenase digestion can be used to effectively isolate ADSCs. ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC, and ChABC expression significantly enhances the migratory capacity of ADSCs. PMID:27364797

  14. BcMctA, a putative monocarboxylate transporter, is required for pathogenicity in Botrytis cinerea.

    Cui, Zhifeng; Gao, Nana; Wang, Qian; Ren, Yun; Wang, Kun; Zhu, Tingheng

    2015-11-01

    Monocarboxylate transporters have a central role in mammalian metabolism, but rarely reported in phytopathogenic fungi. In this study, a putative monocarboxylate transporter gene in Botrytis cinerea [B. cinerea MctA (BcMctA)] was identified in the research of a B. cinerea transfer DNA (T-DNA) insertional mutant (74). Disruption of the gene decreased the growth rate on the medium with monocarboxylate (acetate or pyruvate) as the sole carbon sources, but not affected on lactate. The pyruvate contents in BcmctA deletion mutants decreased about 35 % compared with the wild strain. Besides, the conidial yield was increased about two times in BcmctA disruption mutant. The pathogenicity assay indicated that disruption of BcmctA significantly reduced the virulence of B. cinerea on cucumber and tomato leaves. Our results demonstrated that BcMctA is related to pyruvate uptake and pathogenicity of B. cinerea on cucumber and tomato leaves. PMID:25634672

  15. Review and assessment of package requirements (yellowcake) and emergency response to transportation accidents

    As a consequence of an accident involving a truck shipment of yellowcake, a joint NRC--DOT study was undertaken to review and assess the regulations and practices related to package integrity and to emergency response to transportation accidents involving low specific activity radioactive materials. Recommendations are made regarding the responsibilities of state and local agencies, carriers, and shippers, and the DOT and NRC regulations

  16. Adult active transport in the Netherlands: an analysis of its contribution to physical activity requirements.

    Elliot Fishman

    Full Text Available Modern, urban lifestyles have engineered physical activity out of everyday life and this presents a major threat to human health. The Netherlands is a world leader in active travel, particularly cycling, but little research has sought to quantify the cumulative amount of physical activity through everyday walking and cycling.Using data collected as part of the Dutch National Travel Survey (2010 - 2012, this paper determines the degree to which Dutch walking and cycling contributes to meeting minimum level of physical activity of 150 minutes of moderate intensity aerobic activity throughout the week. The sample includes 74,465 individuals who recorded at least some travel on the day surveyed. As physical activity benefits are cumulative, all walking and cycling trips are analysed, including those to and from public transport. These trips are then converted into an established measure of physical activity intensity, known as metabolic equivalents of tasks. Multivariate Tobit regression models were performed on a range of socio-demographic, transport resources, urban form and meteorological characteristics.The results reveal that Dutch men and women participate in 24 and 28 minutes of daily physical activity through walking and cycling, which is 41% and 55% more than the minimum recommended level. It should be noted however that some 57% of the entire sample failed to record any walking or cycling, and an investigation of this particular group serves as an important topic of future research. Active transport was positively related with age, income, bicycle ownership, urban density and air temperature. Car ownership had a strong negative relationship with physically active travel.The results of this analysis demonstrate the significance of active transport to counter the emerging issue of sedentary lifestyle disease. The Dutch experience provides other countries with a highly relevant case study in the creation of environments and cultures that

  17. Apical Transport of Influenza A Virus Ribonucleoprotein Requires Rab11-positive Recycling Endosome

    Momose, Fumitaka; Sekimoto, Tetsuya; Ohkura, Takashi; Jo, Shuichi; Kawaguchi, Atsushi; Nagata, Kyosuke; Morikawa, Yuko

    2011-01-01

    Influenza A virus RNA genome exists as eight-segmented ribonucleoprotein complexes containing viral RNA polymerase and nucleoprotein (vRNPs). Packaging of vRNPs and virus budding take place at the apical plasma membrane (APM). However, little is known about the molecular mechanisms of apical transport of newly synthesized vRNP. Transfection of fluorescent-labeled antibody and subsequent live cell imaging revealed that punctate vRNP signals moved along microtubules rapidly but intermittently i...

  18. Assessment of Quality Assurance Measures for Radioactive Material Transport Packages not Requiring Competent Authority Design Approval - 13282

    Komann, Steffen; Groeke, Carsten; Droste, Bernhard [BAM Federal Institute for Materials Research and Testing, Unter den Eichen 44-46, 12203 Berlin (Germany)

    2013-07-01

    The majority of transports of radioactive materials are carried out in packages which don't need a package design approval by a competent authority. Low-active radioactive materials are transported in such packages e.g. in the medical and pharmaceutical industry and in the nuclear industry as well. Decommissioning of NPP's leads to a strong demand for packages to transport low and middle active radioactive waste. According to IAEA regulations the 'non-competent authority approved package types' are the Excepted Packages and the Industrial Packages of Type IP-1, IP-2 and IP-3 and packages of Type A. For these types of packages an assessment by the competent authority is required for the quality assurance measures for the design, manufacture, testing, documentation, use, maintenance and inspection (IAEA SSR 6, Chap. 306). In general a compliance audit of the manufacturer of the packaging is required during this assessment procedure. Their regulatory level in the IAEA regulations is not comparable with the 'regulatory density' for packages requiring competent authority package design approval. Practices in different countries lead to different approaches within the assessment of the quality assurance measures in the management system as well as in the quality assurance program of a special package design. To use the package or packaging in a safe manner and in compliance with the regulations a management system for each phase of the life of the package or packaging is necessary. The relevant IAEA-SSR6 chap. 801 requires documentary verification by the consignor concerning package compliance with the requirements. (authors)

  19. Assessment of Quality Assurance Measures for Radioactive Material Transport Packages not Requiring Competent Authority Design Approval - 13282

    The majority of transports of radioactive materials are carried out in packages which don't need a package design approval by a competent authority. Low-active radioactive materials are transported in such packages e.g. in the medical and pharmaceutical industry and in the nuclear industry as well. Decommissioning of NPP's leads to a strong demand for packages to transport low and middle active radioactive waste. According to IAEA regulations the 'non-competent authority approved package types' are the Excepted Packages and the Industrial Packages of Type IP-1, IP-2 and IP-3 and packages of Type A. For these types of packages an assessment by the competent authority is required for the quality assurance measures for the design, manufacture, testing, documentation, use, maintenance and inspection (IAEA SSR 6, Chap. 306). In general a compliance audit of the manufacturer of the packaging is required during this assessment procedure. Their regulatory level in the IAEA regulations is not comparable with the 'regulatory density' for packages requiring competent authority package design approval. Practices in different countries lead to different approaches within the assessment of the quality assurance measures in the management system as well as in the quality assurance program of a special package design. To use the package or packaging in a safe manner and in compliance with the regulations a management system for each phase of the life of the package or packaging is necessary. The relevant IAEA-SSR6 chap. 801 requires documentary verification by the consignor concerning package compliance with the requirements. (authors)

  20. PRÁTICAS DE RESPONSABILIDADE SOCIAL EMPRESARIAL INTERNA EM PEQUENAS EMPRESAS DO GRANDE ABC PAULISTA

    MATTIOLI, José Wiliam

    2015-05-01

    Full Text Available Corporate Social Responsibility is one of the highlights in discussions about the role of business and its commitment to the diverse audiences that relates. This theme has demanded attention and efforts of companies and different sectors of society and government. This requires a design that Social Responsibility beyond what the law requires, and companies that act responsibly with all its stakeholders. One aspect of social commitment refers to the workforce as a competitive advantage in business process management. Thus, this study aims to identify whether small businesses in the industrial sector, develop internal practices of Corporate Social Responsibility. The work is predominantly qualitative, using descriptive held in companies located in the Greater ABC region, selected from the ABCD guide. For both structured questionnaire was used, adapted from data collection instrument developed by the Ethos Institute for Corporate Social Responsibility in partnership with the SEBRAE. The questionnaire was administered in 14 companies in the Greater ABC's industrial sector. The results showed that the surveyed companies, mostly internal social responsibility practices, especially practices that do not require large investments or are determined by law. Other companies that lack such practices are in the process of awareness. A Responsabilidade Social Empresarial (RSE representa um dos destaques das discussões sobre o papel das empresas e seu compromisso com os diversos públicos. O tema tem despertado atenção das empresas de diferentes setores da sociedade e do governo implicando, desta forma, numa concepção de Responsabilidade Social para além do que determina a lei, levando-as a atuar de maneira responsável com todos seus stakeholders. Um dos aspectos do compromisso social se refere ao público interno como um diferencial competitivo das empresas no processo de gestão. Desta forma, o presente trabalho tem como objetivo identificar se

  1. On Application of ABC Approach in Management of Agri- scientific Research Cost

    Liangchun; ZHA

    2014-01-01

    This paper firstly introduced theoretical background of the Activity-Based Costing(ABC). Then,it analyzed necessity,extension resistance and difficulty of ABC approach in agri-scientific research institutions. Finally,it came up with some recommendations for scientifically learning and steadily promoting ABC method in agri-scientific research field.

  2. On Application of ABC Approach in Management of Agri-scientific Research Cost

    Zha, Liangchun

    2014-01-01

    This paper firstly introduced theoretical background of the Activity-Based Costing (ABC). Then, it analyzed necessity, extension resistance and difficulty of ABC approach in agri-scientific research institutions. Finally, it came up with some recommendations for scientifically learning and steadily promoting ABC method in agri-scientific research field.

  3. Regional, national and international security requirements for the transport of nuclear cargo by sea

    Booker, P.A.; Barnwell, I. [Marine Operations, BNFL International Transport and British Nuclear Group Security (United Kingdom)

    2004-07-01

    Since the beginning of the nuclear age in the 1940's, the world has focused on the immense possibilities of nuclear power with both its destructive and productive capabilities. The civil nuclear industry in the UK, as in most nuclear weapons states, grew from the military facilities built in the post war years under the political climate of the Cold War. In the early years of the industry, civil and defence nuclear facilities were inextricably linked both in public perceptions and the regulatory infrastructure under which they operated. The nuclear arms race and the spread of communism overshadowed people's perceptions of there being two separate uses of nuclear material. This was a double edged sword which initially allowed the industry to develop largely unhindered by public concerns but latterly meant the industry could not break away from its roots and to many is still perceived as a dangerous and destructive force. Regulatory frameworks governing all aspects of the industry have developed both nationally and internationally driven by valid public concerns, political agendas and an international consensus that the unregulated use of nuclear material has catastrophic possibilities on an international scale. With the internationalisation of the civil nuclear industry and the costs associated with developing facilities to fully support each stage of the fuel cycle, from enrichment, fuel manufacturing, reprocessing and waste remediation, it became inevitable that a transport infrastructure would develop to make best use of the facilities. Regulations, both national and international are implicit in ensuring the security of nuclear material in transit. Due to the physical size of many of the irradiated fuel packages and implications of the changes to transport safety regulations, international transports of nuclear material, other than within mainland Europe, is predominantly carried out by sea.

  4. Siting of an MRS facility: identification of a geographic region that reduces transportation requirements

    The study reported here was undertaken as part of the site screening and evaluation activities for the Monitored Retrievable Storage (MRS) Program of the Office of Civilian Radioactive Waste Management (OCRWM), Department of Energy (DOE). Its primary purpose was to determine: the location and shape of a preferred geographic region within which locating an MRS facility would minimize total shipment miles for spent fuel transported through the MRS facility to a repository, and the sensitivity of the location and shape of this region and the reduction in total shipment miles to possible variations in waste management system logistics. As a result of this analysis, a geographic region has been identified which is preferred for siting an MRS facility. This region will be referred to as the preferred region in this study. Siting an MRS facility in the preferred region will limit total shipment miles (i.e., the total miles traveled for all shipments of spent fuel) to and from the MRS facility to within 20% of the lowest achievable. The region is preferred for a mixed truck/rail system of transport from reactors to the MRS facility. It is assumed that rail will be used to ship spent fuel from the MRS facility to a geologic repository for disposal. Siting an MRS facility in the preferred region will reduce total shipment miles for all currently considered system logistics options which include an MRS facility in the system. These options include: any first repository location, the possible range of spent fuel consolidation at the MRS, use of multi-cask or single-cask train shipments, use of current or future spent fuel transport casks, servicing only the first or both the first and second repositories, and shipment of fuel from western reactors either through the MRS facility or to a western facility (a second, smaller MRS facility or the first repository)

  5. Regional, national and international security requirements for the transport of nuclear cargo by sea

    Since the beginning of the nuclear age in the 1940's, the world has focused on the immense possibilities of nuclear power with both its destructive and productive capabilities. The civil nuclear industry in the UK, as in most nuclear weapons states, grew from the military facilities built in the post war years under the political climate of the Cold War. In the early years of the industry, civil and defence nuclear facilities were inextricably linked both in public perceptions and the regulatory infrastructure under which they operated. The nuclear arms race and the spread of communism overshadowed people's perceptions of there being two separate uses of nuclear material. This was a double edged sword which initially allowed the industry to develop largely unhindered by public concerns but latterly meant the industry could not break away from its roots and to many is still perceived as a dangerous and destructive force. Regulatory frameworks governing all aspects of the industry have developed both nationally and internationally driven by valid public concerns, political agendas and an international consensus that the unregulated use of nuclear material has catastrophic possibilities on an international scale. With the internationalisation of the civil nuclear industry and the costs associated with developing facilities to fully support each stage of the fuel cycle, from enrichment, fuel manufacturing, reprocessing and waste remediation, it became inevitable that a transport infrastructure would develop to make best use of the facilities. Regulations, both national and international are implicit in ensuring the security of nuclear material in transit. Due to the physical size of many of the irradiated fuel packages and implications of the changes to transport safety regulations, international transports of nuclear material, other than within mainland Europe, is predominantly carried out by sea

  6. I am Calm: Towards a Psychoneurological Evaluation of ABC Ringtones

    John N.A. Brown

    2015-10-01

    Full Text Available Anthropology-Based Computing (ABC suggests that socio-cultural, neurological, and physiological parameters of normal human interaction with the world can be applied to current technology in order to improve Human-Computer Interaction (HCI. To challenge this theory, we hypothesized smartphone ringtones that could be targeted to specific people in a manner that would inform them without disturbing their work or the work of others. In this paper we report the quantitative data from the first formal trials of these ‘ABC ringtones’. Beta Wave activity patterns were recorded in the brains of 10 participants exposed to 5 different ringtones at three different volumes while they were focused on performing a typing test in a noisy environment. Our preliminary findings seem to show that the ABC ringtones - at a volume too low to be consciously heard - triggered a response in the pre-attentive part of the brain, and that the embedded information was transferred to the attentive part of the brain by an internal mechanism that did not disrupt the work being done in the typing task. We propose that these results provide preliminary evidence for the ABC model of HCI and its explanation of the centering mechanism that is requisite if Peripheral Interaction is to be applied in changing Ubiquitous Computing into Calm Technology.

  7. Dynamics of 'abc' and 'qd' constant parameters induction generator model

    Fajardo-R, L.A.; Medina, A.; Iov, F.

    2009-01-01

    In this paper, parametric sensibility effects on dynamics of the induction generator in the presence of local perturbations are investigated. The study is conducted in a 3x2 MW wind park dealing with abc, qd0 and qd reduced order, induction generator model respectively, and with fluxes as state...

  8. Selections from the ABC 2012 Annual Convention, Honolulu, Hawaii

    Whalen, D. Joel

    2013-01-01

    The 13 Favorite Assignments featured here were presented at the 2012 Association for Business Communication (ABC) Annual Convention, Honolulu, Hawaii. A variety of learning objectives are featured, including the following: enhancing resume's visual impact, interpersonal skills, social media, team building, web design, community service…

  9. The ABCs for Pre-Service Teacher Cultural Competency Development

    He, Ye; Cooper, Jewell E.

    2009-01-01

    In an effort to combine pre-service teachers' self-reflection with their field experiences to enhance their cultural competency, this study adopted Schmidt's ABC's (Autobiography, Biography, and Cross-cultural Comparison) Model in two courses in a pre-service teacher education program. Through group comparisons, this study measured the impact that…

  10. Preconceptual ABC design definition and system configuration layout

    This document is the conceptual design document for the follow-on to the Molten Salt Breeder Reactor, known as the ABC type reactor. It addresses blanket design options, containment options, off-gas systems, drainage systems, and components/layouts of the primary, secondary, and tertiary systems, and it contains a number of diagrams for the configuration of the major systems

  11. Conceptual proposals on safeguards while implementation of ABC-project

    The report deals with some conceptual problems of nuclear safeguard measures in connection with project ABC implementation. Safeguards issues in this area have a lot of common features with traditional approaches at existing facilities, at the same time there are some characteristic features arising in connection with specific technology process in this project

  12. Analysis of operational requirements for medium density air transportation, volume 2

    1975-01-01

    The medium density air travel market is examined and defined in terms of numbers of people transported per route per day and frequency of service. The operational characteristics for aircraft to serve this market are determined and a basepoint aircraft is designed from which tradeoff studies and parametric variations can be conducted. The impact of the operational characteristics on the air travel system is evaluated along with the economic viability of the study aircraft. Research and technology programs for future study consideration are identified.

  13. Localization of Iron in Arabidopsis Seed Requires the Vacuolar Membrane Transporter VIT1

    Iron deficiency is a major human nutritional problem wherever plant-based diets are common. Using synchrotron x-ray fluorescence microtomography to directly visualize iron in Arabidopsis seeds, we show that iron is localized primarily to the provascular strands of the embryo. This localization is completely abolished when the vacuolar iron uptake transporter VIT1 is disrupted. Vacuolar iron storage is also critical for seedling development because vit1-1 seedlings grow poorly when iron is limiting. We have uncovered a fundamental aspect of seed biology that will ultimately aid the development of nutrient-rich seed, benefiting both human health and agricultural productivity

  14. SGLT1 sugar transporter/sensor is required for post-oral glucose appetition.

    Sclafani, Anthony; Koepsell, Hermann; Ackroff, Karen

    2016-04-01

    Recent findings suggest that the intestinal sodium-glucose transporter 1 (SGLT1) glucose transporter and sensor mediates, in part, the appetite-stimulation actions of intragastric (IG) glucose and nonmetabolizable α-methyl-d-glucopyranoside (MDG) infusions in mice. Here, we investigated the role of SGLT1 in sugar conditioning using SGLT1 knockout (KO) and C57BL/6J wild-type (WT) mice. An initial experiment revealed that both KO and WT mice maintained on a very low-carbohydrate diet display normal preferences for saccharin, which was used in the flavored conditioned stimulus (CS) solutions. In experiment 2, mice were trained to drink one flavored solution (CS+) paired with an IG MDG infusion and a different flavored solution (CS-) paired with IG water infusion. In contrast to WT mice, KO mice decreased rather than increased the intake of the CS+ during training and failed to prefer the CS+ over the CS- in a choice test. In experiment 3, the KO mice also decreased their intake of a CS+ paired with IG glucose and avoided the CS+ in a choice test, unlike WT mice, which preferred the CS+ to CS-. In experiment 4, KO mice, like WT mice preferred a glucose + saccharin solution to a saccharin solution. These findings support the involvement of SGLT1 in post-oral glucose and MDG conditioning. The results also indicate that sugar malabsorption in KO mice has inhibitory effects on sugar intake but does not block their natural preference for sweet taste. PMID:26791832

  15. Cross sections for electron and photon processes required by electron-transport calculations

    Electron-transport calculations rely on a large collection of electron-atom and photon-atom cross-section data to represent the response characteristics of the target medium. These basic atomic-physics quantities, and certain qualities derived from them that are now commonly in use, are critically reviewed. Publications appearing after 1978 are not given consideration. Processes involving electron or photon energies less than 1 keV are ignored, while an attempt is made to exhaustively cover the remaining independent parameters and target possibilities. Cases for which data improvements can be made from existing information are identified. Ranges of parameters for which state-of-the-art data are not available are sought out, and recommendations for explicit measurements and/or calculations with presently available tools are presented. An attempt is made to identify the maturity of the atomic-physics data and to predict the possibilities for rapid changes in the quality of the data. Finally, weaknesses in the state-of-the-art atomic-physics data and in the conceptual usage of these data in the context of electron-transport theory are discussed. Brief attempts are made to weight the various aspects of these questions and to suggest possible remedies

  16. Survey of Packaging Requirements for the Transport of Highly Hazardous Materials

    Among hazardous materials those which are most dangerous fall into three categories: chemical, biological, and radioactive. The DOT hazard classes for these three categories are Hazard Class 2.3 (poisonous gases) and 6.1 (toxic substances) for chemical hazards, Hazard Class 6.2 (infectious substances) for biological hazards and Hazard Class 7 for radioactive material (RAM) hazards. The packaging requirements for chemical and biological hazards are outlined and compared with RAM packaging requirements. RAM packages are found to be able to withstand much more severe performance tests than packages for other, more lethal hazards

  17. The role of half-transporters in multidrug resistance

    Bates, S E; Robey, R; Miyake, K;

    2001-01-01

    role of drug transporters in clinical drug resistance. These newly identified transporters include additional members of the MRP family, ABC2, and a new half-transporter, MXR/BCRP/ABCP1. This half-transporter confers high levels of resistance to mitoxantrone, anthracyclines, and the camptothecins SN-38...

  18. 40 CFR 51.309 - Requirements related to the Grand Canyon Visibility Transport Commission.

    2010-07-01

    ... compliance with air-pollution related requirements. (iii) Programs to preserve and expand energy conservation... associated with the renewable energy goals, energy efficiency and pollution prevention activities. (vi) A... purposes of 40 CFR 51.309, means electricity generated by non-nuclear and non-fossil low or no air...

  19. 49 CFR 171.23 - Requirements for specific materials and packagings transported under the ICAO Technical...

    2010-10-01

    ... relief devices and provide appropriate guidance for exposure to fire. (b) Conditions and requirements..., or a DOT exemption or special permit cylinder or a cylinder used as a fire extinguisher in... properly marked valve; and (iv) A UN pressure receptacle is marked with “USA” as a country of approval...

  20. 76 FR 79544 - Processing of Merchant Mariner Credentials for Those Mariners Not Requiring a Transportation...

    2011-12-22

    ... SECURITY Coast Guard 46 CFR Parts 10, 11, 12, and 15 Processing of Merchant Mariner Credentials for Those... acquire and renew a Merchant Mariner Credential (MMC) without holding a valid TWIC. DATES: This policy is... regulations require every applicant for an original or renewal of a Merchant Mariner Credential (MMC) obtain...

  1. 75 FR 43906 - Hazardous Materials: Requirements for the Storage of Explosives During Transportation

    2010-07-27

    ... appropriate safety measures, because it assesses the unique characteristics of a particular site. DDESB... Fire Protection Association (NFPA) standard 498 as the Federally approved standard for the construction... rulemaking under Docket HM-232A (67 FR 46622) entitled ``Security Requirements for Motor...

  2. A Compute Environment of ABC95 Array Computer Based on Multi-FPGA Chip

    2000-01-01

    ABC95 array computer is a multi-function network's computer based on FPGA technology, The multi-function network supports processors conflict-free access data from memory and supports processors access data from processors based on enhanced MESH network.ABC95 instruction's system includes control instructions, scalar instructions, vectors instructions.Mostly net-work instructions are introduced.A programming environment of ABC95 array computer assemble language is designed.A programming environment of ABC95 array computer for VC++ is advanced.It includes load function of ABC95 array computer program and data, store function, run function and so on.Specially, The data type of ABC95 array computer conflict-free access is defined.The results show that these technologies can develop programmer of ABC95 array computer effectively.

  3. Help guide for the application of regulatory requirements on the transport of radioactive material; Guia de ayuda para la aplicacion de requisitos reglementarios sobr el transporte de material radiactivo

    Martin Rodriguez, S.; Acena Moreno, V.; Zamora Martin, F.; Rubio de Juan, E.

    2011-07-01

    The regulation of transport of radioactive material by road in Spain refers to compliance with the requirements of the European Agreement concerning the International Carriage of Dangerous Goods by Road (ADR) in force. The structure presented by this legislation, which is international, and the inclusion in it of requirements that apply to other dangerous goods makes it difficult to consult the requirements that specifically apply to the transport of radioactive material. Therefore, the Nuclear Safety Council (CSN) has found it necessary to publish a guide that facilitates users to comply with its provisions and, consequently, this transport security.

  4. A Salmonella protein that is required for resistance to antimicrobial peptides and transport of potassium.

    Parra-Lopez, C; Lin, R; Aspedon, A; Groisman, E A

    1994-01-01

    The ability of invading pathogens to proliferate within host tissues requires the capacity to resist the killing effects of a wide variety of host defense molecules. sap mutants of the facultative intracellular parasite Salmonella typhimurium exhibit hypersensitivity to antimicrobial peptides, cannot survive within macrophages in vitro and are attenuated for mouse virulence in vivo. We conducted a molecular genetic analysis of the sapG locus and showed that it encodes a product that is 99% id...

  5. A sodium requirement for growth, solute transport, and pH homeostasis in Bacillus firmus RAB.

    Krulwich, T A; Guffanti, A A; Bornstein, R F; Hoffstein, J

    1982-02-25

    Activity of a Na+/H+ antiporter has been suggested to be critically involved in pH homeostasis in obligately alkalophilic bacteria (Krulwich, K. A., Mandel, K. G., Bornstein, R. F., and Guffanti, A. A. (1979) Biochem. Biophys. Res. Commun. 91, 58-62) and in Escherichia coli (Zilberstein, D., Padan, E., and Schuldiner, S. (1980) FEBS Lett. 116, 177-180). A concern with respect to these proposals has been the failure of either Bacillus alcalophilus or E. coli to exhibit a requirement for added Na+ for growth. Thus, it became of interest to examine Na+-coupled porter functions in obligately alkalophilic Bacillus firmus RAB, a species that exhibits an absolute requirement for added Na+ for growth at pH 10.5. In a comparative study using membrane vesicles from B. alcalophilus and B. firmus RAB it was found that both the Na+/H+ antiporter and the Na+/alpha-aminoisobutyric acid symporter from the "Na+-requiring" species had much lower apparent affinities for Na+ than corresponding porters from B. alcalophilus. At high concentrations of Na+, the porters from the two species were functionally similar. These findings support the argument that the absence of a growth requirement for added Na+ may reflect an ability of at least some bacteria to effectively utilize and recycle the available levels of Na+ that contaminate all media, rather than reflect true Na+ independence. Studies with a nonalkalophilic derivative of B. firmus RAB confirmed earlier findings with B. alcalophilus of a pleiotropic loss of Na+ coupling to porters in nonalkalophilic mutants. PMID:7056750

  6. A Feasibility Study for Gamification in Transport Maintenance : Requirements to implement gamification in heterogeneous organizations

    Shreenath, Vinutha Magal; Kornevs, Maksims; Raghothama, Jayanth; Meijer, Sebastiaan

    2015-01-01

    Gamification has been successfully applied in many domains, but mostly for simple, isolated and operational tasks. The hope for gamification as a method to radically change and improve behavior, to provide incentives for sustained engagement has proven to be more difficult to get right. Applying gamification in large networked organizations with heterogeneous tasks remains a challenge. Applying gamification in such enterprise environments posits different requirements, and a match between the...

  7. Potential role of ABC-assisted repositories in U.S. plutonium and high-level waste disposition

    This paper characterizes the issues involving deep geologic disposal of LWR spent fuel rods, then presents results of an investigation to quantify the potential role of Accelerator-Based Conversion (ABC) in an integrated national nuclear materials and high level waste disposition strategy. The investigation used the deep geological repository envisioned for Yucca Mt., Nevada as a baseline and considered complementary roles for integrated ABC transmutation systems. The results indicate that although a U.S. geologic waste repository will continue to be required, waste partitioning and accelerator transmutation of plutonium, the minor actinides, and selected long-lived fission products can result in the following substantial benefits: plutonium burndown to near zero levels, a dramatic reduction of the long term hazard associated with geologic repositories, an ability to place several-fold more high level nuclear waste in a single repository, electricity sales to compensate for capital and operating costs

  8. Double pionic fusion. Towards an understanding of the ABC puzzle by exclusive measurements

    Bashkanov, M.

    2006-07-01

    The ABC effect is a huge unexpected enhancement at twice the pion mass in the invariant mass spectrum of two pions, which are generated in double-pionic fusion to bound nuclear systems. This peculiar phenomenon has been missing a conclusive explanation all the time since it has been discovered 1960 in single-arm measurements of {sup 3}He ejectiles in the reaction pd{yields} {sup 3}HeX. One reason for this failure has been that all measurements to this subject have been inclusive, i.e., lacking the full experimentally accessible information. Hence exclusive measurements were performed at CELSIUS/WASA at an energy of T{sub p}=0.895 GeV, where the ABC effect is expected to be strongest. For the first time exclusive data of solid statistics for both the pd{yields}{sup 3}He{pi}{sup 0}{pi}{sup 0} and pd{yields}{sup 3}He{pi}{sup +}{pi}{sup -} reactions were obtained including also results for the three-pion production total cross-section. The new data are consistent with the previous inclusive data. They provide, however, much more additional information, which rule out all previous explications of the ABC effect. The now available kinematically complete set of data reveals that the low {pi}{pi}-mass enhancement (ABC-effect): - is not necessarily associated with a high {pi}{pi}-mass enhancement, - is always connected with the simultaneous excitation of two {delta} resonances, - is of scalar-isoscalar nature, i.e. a {sigma}-channel phenomenon, - requires dynamics in the reaction system, which has not been considered hitherto. Various possible solutions are discussed, however, all of them demand a high attraction in the {delta}{delta} system - a point, which has never been touched so far in theoretical and experimental investigations. For this data analysis new powerful methods based on neural nets have been developed. Their current and possible future applications are discussed. (orig.)

  9. Historical background of the development of various requirements in the international regulations for the safe packaging and transport of radioactive material

    Questions are frequently asked regarding the source of some of the package test requirements in the Transport Regulations, the philosophy behind them and the basis for selecting them. This paper summarizes the results of a review of early historical documents and elaborates on the early philosophy behind the regulatory requirements. To the extent possible, the paper compares the early philosophy with the current structure of the Transport Regulations in key topic areas with a focus on the test requirements for packages that are designed to withstand accident conditions of transport

  10. myo-Inositol-1-phosphate synthase is required for polar auxin transport and organ development

    Chen, Hao

    2010-06-01

    myo-Inositol-1-phosphate synthase is a conserved enzyme that catalyzes the first committed and rate-limiting step in inositol biosynthesis. Despite its wide occurrence in all eukaryotes, the role of myo-inositol-1-phosphate synthase and de novo inositol biosynthesis in cell signaling and organism development has been unclear. In this study, we isolated loss-of-function mutants in the Arabidopsis MIPS1 gene from different ecotypes. It was found that all mips1 mutants are defective in embryogenesis, cotyledon venation patterning, root growth, and root cap development. The mutant roots are also agravitropic and have reduced basipetal auxin transport. mips1 mutants have significantly reduced levels of major phosphatidylinositols and exhibit much slower rates of endocytosis. Treatment with brefeldin A induces slower PIN2 protein aggregation in mips1, indicating altered PIN2 trafficking. Our results demonstrate that MIPS1 is critical for maintaining phosphatidylinositol levels and affects pattern formation in plants likely through regulation of auxin distribution. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Requirements for Forming Efficient 3-D Charge Transport Pathway in Diketopyrrolopyrrole-Based Copolymers: Film Morphology vs Molecular Packing.

    Lee, Gang-Young; Han, A-Reum; Kim, Taewan; Lee, Hae Rang; Oh, Joon Hak; Park, Taiho

    2016-05-18

    To achieve extremely high planarity and processability simultaneously, we have newly designed and synthesized copolymers composed of donor units of 2,2'-(2,5-dialkoxy-1,4-phenylene)dithieno[3,2-b]thiophene (TT-P-TT) and acceptor units of diketopyrrolopyrrole (DPP). These copolymers consist of a highly planar backbone due to intramolecular interactions. We have systematically investigated the effects of intermolecular interactions by controlling the side chain bulkiness on the polymer thin-film morphologies, packing structures, and charge transport. The thin-film microstructures of the copolymers are found to be critically dependent upon subtle changes in the intermolecular interactions, and charge transport dynamics of the copolymer based field-effect transistors (FETs) has been investigated by in-depth structure-property relationship study. Although the size of the fibrillar structures increases as the bulkiness of the side chains in the copolymer increases, the copolymer with the smallest side chain shows remarkably high charge carrier mobility. Our findings reveal the requirement for forming efficient 3-D charge transport pathway and highlight the importance of the molecular packing and interdomain connectivity, rather than the crystalline domain size. The results obtained herein demonstrate the importance of tailoring the side chain bulkiness and provide new insights into the molecular design for high-performance polymer semiconductors. PMID:27117671

  12. Blood-Brain Barrier Active Efflux Transporters: ATP-Binding Cassette Gene Family

    Löscher, Wolfgang; Potschka, Heidrun

    2005-01-01

    Summary: The blood-brain barrier (BBB) contributes to brain homeostasis by protecting the brain from potentially harmful endogenous and exogenous substances. BBB active drug efflux transporters of the ATP-binding cassette (ABC) gene family are increasingly recognized as important determinants of drug distribution to, and elimination from, the CNS. The ABC efflux transporter P-glycoprotein (Pgp) has been demonstrated as a key element of the BBB that can actively transport a huge variety of lip...

  13. ABCA Transporter Gene Expression and Poor Outcome in Epithelial Ovarian Cancer

    Hedditch, Ellen L; Gao, Bo; Russell, Amanda J;

    2014-01-01

    BACKGROUND: ATP-binding cassette (ABC) transporters play various roles in cancer biology and drug resistance, but their association with outcomes in serous epithelial ovarian cancer (EOC) is unknown. METHODS: The relationship between clinical outcomes and ABC transporter gene expression in two...... cancer cell growth and migration in vitro, and statin treatment reduced ovarian cancer cell migration. CONCLUSIONS: Expression of ABCA transporters was associated with poor outcome in serous ovarian cancer, implicating lipid trafficking as a potentially important process in EOC....

  14. Examination of the combined effects of chondroitinase ABC, growth factors and locomotor training following compressive spinal cord injury on neuroanatomical plasticity and kinematics.

    Olivier Alluin

    Full Text Available While several cellular and pharmacological treatments have been evaluated following spinal cord injury (SCI in animal models, it is increasingly recognized that approaches to address the glial scar, including the use of chondroitinase ABC (ChABC, can facilitate neuroanatomical plasticity. Moreover, increasing evidence suggests that combinatorial strategies are key to unlocking the plasticity that is enabled by ChABC. Given this, we evaluated the anatomical and functional consequences of ChABC in a combinatorial approach that also included growth factor (EGF, FGF2 and PDGF-AA treatments and daily treadmill training on the recovery of hindlimb locomotion in rats with mid thoracic clip compression SCI. Using quantitative neuroanatomical and kinematic assessments, we demonstrate that the combined therapy significantly enhanced the neuroanatomical plasticity of major descending spinal tracts such as corticospinal and serotonergic-spinal pathways. Additionally, the pharmacological treatment attenuated chronic astrogliosis and inflammation at and adjacent to the lesion with the modest synergistic effects of treadmill training. We also observed a trend for earlier recovery of locomotion accompanied by an improvement of the overall angular excursions in rats treated with ChABC and growth factors in the first 4 weeks after SCI. At the end of the 7-week recovery period, rats from all groups exhibited an impressive spontaneous recovery of the kinematic parameters during locomotion on treadmill. However, although the combinatorial treatment led to clear chronic neuroanatomical plasticity, these structural changes did not translate to an additional long-term improvement of locomotor parameters studied including hindlimb-forelimb coupling. These findings demonstrate the beneficial effects of combined ChABC, growth factors and locomotor training on the plasticity of the injured spinal cord and the potential to induce earlier neurobehavioral recovery. However

  15. ABC: software for interactive browsing of genomic multiple sequence alignment data

    Singaravelu Senthil AG

    2004-12-01

    Full Text Available Abstract Background Alignment and comparison of related genome sequences is a powerful method to identify regions likely to contain functional elements. Such analyses are data intensive, requiring the inclusion of genomic multiple sequence alignments, sequence annotations, and scores describing regional attributes of columns in the alignment. Visualization and browsing of results can be difficult, and there are currently limited software options for performing this task. Results The Application for Browsing Constraints (ABC is interactive Java software for intuitive and efficient exploration of multiple sequence alignments and data typically associated with alignments. It is used to move quickly from a summary view of the entire alignment via arbitrary levels of resolution to individual alignment columns. It allows for the simultaneous display of quantitative data, (e.g., sequence similarity or evolutionary rates and annotation data (e.g. the locations of genes, repeats, and constrained elements. It can be used to facilitate basic comparative sequence tasks, such as export of data in plain-text formats, visualization of phylogenetic trees, and generation of alignment summary graphics. Conclusions The ABC is a lightweight, stand-alone, and flexible graphical user interface for browsing genomic multiple sequence alignments of specific loci, up to hundreds of kilobases or a few megabases in length. It is coded in Java for cross-platform use and the program and source code are freely available under the General Public License. Documentation and a sample data set are also available http://mendel.stanford.edu/sidowlab/downloads.html.

  16. ABC Tester - Artificial Bee Colony Based Software Test Suite Optimization Approach

    D. Jeya Mala

    2009-07-01

    Full Text Available In this paper we present a new, non-pheromone-based test suite optimization approach inspired by the behavior of biological bees. Our proposed approach is based on ABC (Artificial Bee Colony Optimization which is motivated by the intelligent behavior of honey bees. In our proposed system, the sites are the nodes in the Software under Test (SUT, the artificial bees modify the test cases with time and the bee?s aim is to discover the places of nodes with higher coverage and finally the one with the highest usage by the given test case. Since ABC system combines local search methods carried out by employed bees with global search methods managed by onlookers and scouts, we attain near global optima. We investigate whether this new approach outperforms existing test optimization approach based on Genetic Algorithms (GA in the task of software test optimization. Taking into account the results of our experiments, we conclude that (i the proposed approach uses fewer iterations to complete the task; (ii is more scalable, i.e., it requires less computation time to complete the task, and finally (iii our approach is best in achieving near global optimal solution.

  17. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    Zhang Jianzhen

    2011-10-01

    Full Text Available Abstract Background The ATP-binding cassette (ABC transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight subfamilies (A-H by phylogenetic analysis. Gene duplication is very evident in the ABCC and ABCG subfamilies, whereas gene numbers and structures are well conserved in the ABCD, ABCE, ABCF, and ABCH subfamilies. Microarray analysis revealed that expression of 32 silkworm ABC genes can be detected in at least one tissue during different developmental stages, and the expression patterns of some of them were confirmed by quantitative real-time PCR. A large number of ABC genes were highly expressed in the testis compared to other tissues. One of the ABCG genes, BmABC002712, was exclusively and abundantly expressed in the Malpighian tubule implying that BmABC002712 plays a tissue-specific role. At least 5 ABCG genes, including BmABC005226, BmABC005203, BmABC005202, BmABC010555, and BmABC010557, were preferentially expressed in the midgut, showing similar developmental expression profiles to those of 20-hydroxyecdysone (20E-response genes. 20E treatment induced the expression of these ABCG genes in the midgut and RNA interference-mediated knockdown of USP, a component of the 20E receptor, decreased their expression, indicating that these midgut-specific ABCG genes are 20E-responsive. Conclusion In this study, a genome-wide analysis of the silkworm ABC transporters has been conducted. A comparison of ABC transporters from 5 insect species provides an overview of this vital gene superfamily in insects. Moreover, tissue- and stage-specific expression data of the

  18. Model complexities and requirements for multimodal transport network design: Assessment of classical, state-of-the-practice, and state-of-the-research models

    Van Eck, G.; Brands, T.; Wismans, L.J.J.; Pel, A.J.; Van Nes, R.

    2014-01-01

    In the aim for a more sustainable transport system, governments try to stimulate multimodal trip making by facilitating smooth transfers between modes. The assessment of related multimodal policy measures requires transport models that are capable of handling the complex nature of multimodality. Thi

  19. IMPLEMENTATION OF THE ABC COSTING IN A SERVICES PROVIDER COMPANY

    Luiz Ricardo Aguena Jacintho Gil de Castro

    2012-12-01

    Full Text Available This paper describes the implementation of the Activity-Based Cost (ABC method in a company of taxes and accounting services and outlines the positive and negative aspects encountered during implementation. It should be taken into account that this work has been developed in the fiscal area of cost verification. Bibliographical references, internal company documents and interviews with industry officials and the administrator responsible for the company were used. In the evolving of operations, the costs of the fiscal area and their main activities have been described monthly and through this information, drivers have been developed and the ABC (Activity-Based Costing method has been adopted. With the implementation it became clear that the system provides better visualization for the decision making process, it also provided learning for the company, so that the method should be used for an undetermined period of time.

  20. HutZ Is Required for Efficient Heme Utilization in Vibrio cholerae

    Wyckoff, Elizabeth E.; Schmitt, Michael; Wilks, Angela; Shelley M Payne

    2004-01-01

    Vibrio cholerae, the causative agent of cholera, requires iron for growth. One mechanism by which it acquires iron is the uptake of heme, and several heme utilization genes have been identified in V. cholerae. These include three distinct outer membrane receptors, two TonB systems, and an apparent ABC transporter to transfer heme across the inner membrane. However, little is known about the fate of the heme after it enters the cell. In this report we show that a novel heme utilization protein...

  1. A STUDY ON THE ABC APPROACH IN COST MANAGEMENT PRACTICE

    Violeta ISAI; Riana Iren RADU; Camelia IONESCU

    2014-01-01

    Activity-based costing (ABC), an alternative approach to traditional accounting, represents a costing methodology that identifies the main cost drivers, or the main activities in an organization, thus assigning the cost of the products and services according to the number of specific activities or transactions used in the development process of a product or service. This system is based on the measurement of all the activities performed within an organization and provides the companies with t...

  2. IMPLEMENTATION OF THE ABC COSTING IN A SERVICES PROVIDER COMPANY

    Luiz Ricardo Aguena Jacintho Gil de Castro; Fernando de Almeida Santos

    2012-01-01

    This paper describes the implementation of the Activity-Based Cost (ABC) method in a company of taxes and accounting services and outlines the positive and negative aspects encountered during implementation. It should be taken into account that this work has been developed in the fiscal area of cost verification. Bibliographical references, internal company documents and interviews with industry officials and the administrator responsible for the company were used. In the evolving of opera...

  3. Supramolecular Polymers Formed by ABC Miktoarm Star Peptides

    Lin, Yi-an; Ou, Yu-Chuan; Cheetham, Andrew G.; Cui, Honggang

    2013-01-01

    We report here the design and synthesis of an ABC miktoarm star peptide connecting through a lysine junction a short peptide sequence and two hydrophobic but immiscible blocks (a hydrocarbon and a fluorocarbon). The designed molecule can self-assemble into one-dimensional nanostructures with a great diversity of kinetically evolving morphologies in aqueous solution, while molecules that contain only one of the two hydrophobic blocks form structurally similar filaments. We believe the rich ass...

  4. Fuel cells for transport: can the promise be fulfilled? Technical requirements and demands from customers

    Klaiber, Thomas

    The paper discusses the technical requirements and the customer demands for vehicles that have an on-board methanol reformer and fuel cells. The research concentrates on the technical developmental risks which include minimizing volume, reducing weight and, at the same time, improving efficiency and system dynamics. Fuel cell powered vehicles with methanol reformers are not only suitable for a niche market but also these vehicles will compete with conventional vehicles. The greatest hindrance will be the price of the fuel cell. A possible progressive development of the number of fuel cell powered vehicles in conjunction with a reduction in costs will be discussed in the paper. When fuel cell vehicles come to the market it is necessary that an infrastructure for the fuel methanol or hydrogen is installed. Therefore, it will only be possible to introduce fuel cell vehicles into special markets, e.g. California. Such a process will need to be subsidized by additional incentives like tax concessions. Today there are many technical risks and unsolved problems relating to production technologies, infrastructure, and costs. Nevertheless, among the alternative power units, the fuel cell seems to be the only one that might be competitive to the conventional power unit, especially relating to emissions.

  5. Second-order nonlinear optical metamaterials: ABC-type nanolaminates

    Alloatti, L., E-mail: alloatti@mit.edu; Kieninger, C.; Lauermann, M.; Köhnle, K. [Institute of Photonics and Quantum Electronics (IPQ), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); Froelich, A.; Wegener, M. [Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); DFG-Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); Institute of Nanotechnology, Karlsruhe Institute of Technology (KIT), 76021 Karlsruhe (Germany); Frenzel, T. [Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); Freude, W. [Institute of Photonics and Quantum Electronics (IPQ), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); Institute for Microstructure Technology (IMT), Karlsruhe Institute of Technology (KIT), 76344 Eggenstein-Leopoldshafen (Germany); Leuthold, J.; Koos, C., E-mail: christian.koos@kit.edu [Institute of Photonics and Quantum Electronics (IPQ), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); DFG-Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe (Germany); Institute for Microstructure Technology (IMT), Karlsruhe Institute of Technology (KIT), 76344 Eggenstein-Leopoldshafen (Germany)

    2015-09-21

    We demonstrate a concept for second-order nonlinear metamaterials that can be obtained from non-metallic centrosymmetric constituents with inherently low optical absorption. The concept is based on iterative atomic-layer deposition of three different materials, A = Al{sub 2}O{sub 3}, B = TiO{sub 2}, and C = HfO{sub 2}. The centrosymmetry of the resulting ABC stack is broken since the ABC and the inverted CBA sequences are not equivalent—a necessary condition for non-zero second-order nonlinearity. In our experiments, we find that the bulk second-order nonlinear susceptibility depends on the density of interfaces, leading to a nonlinear susceptibility of 0.26 pm/V at a wavelength of 800 nm. ABC-type nanolaminates can be deposited on virtually any substrate and offer a promising route towards engineering of second-order optical nonlinearities at both infrared and visible wavelengths.

  6. ABC Supported Handover Decision Scheme based on multi-PSO with Optimum Mutation

    Xingwei Wang

    2011-08-01

    Full Text Available In this paper, a handover decision scheme with ABC (always best connected supported is proposed. With knowledge of fuzzy mathematics and microeconomics introduced, application types, QoS (quality of service requirements, access networks and terminals are described. With access network conditions, application requirements, user preferences to access network coding schemes, user preferences to access network providers, terminal velocities and terminal battery capacities considered comprehensively, an optimal handover solution of assigning N terminals to M access networks is found based on multi-PSO (particle swarm optimization with optimum mutation. With the help of gaming analysis, Pareto optimum under Nash equilibrium of both user utility and network provider utility is achieved or approached for the found solution. Simulation results showed that it was effective.

  7. ATP-binding cassette transporters as pitfalls in selection of transgenic cells.

    Theile, Dirk; Staffen, Bianca; Weiss, Johanna

    2010-04-15

    Puromycin, hygromycin, and geneticin (G418) are antibiotics frequently used to select genetically engineered eukaryotic cells after transfection or transduction. Because intrinsic or acquired high expression of ATP-binding cassette (ABC) transporters, such as P-glycoprotein (Pgp/ABCB1) and multidrug resistance-associated proteins (MRP/ABCC1), can hamper efficient selection, it is important to know whether these antibiotics are substrates and/or inducers of efflux transporters. Therefore, we investigated the influence of these antibiotics on drug transporter expression by quantitative real-time polymerase chain reaction in the induction model cell line LS180. Moreover, we assessed whether ABC transporters influence the growth inhibitory effects of these antibiotics by proliferation assays using Madin-Darby canine kidney II (MDCKII) cells overexpressing the particular transporter. The results obtained indicate that puromycin and G418 are substrates of several ABC transporters, mainly Pgp/ABCB1. In contrast, hygromycin seems to be no good substrate for any of the ABC transporters investigated. Puromycin induced ABCC1/MRP1, whereas G418 suppressed ABCB1/Pgp, at the messenger RNA (mRNA) level. In contrast, hygromycin had no effect on ABC transporter mRNA expressions. In conclusion, this study emphasizes the significance of ABC transporters for the efficacy of selection processes. Consciousness of the results is supposed to guide the molecular biologist to the right choice of adequate experimental conditions for successful selection of genetically engineered eukaryotic cells. PMID:20018165

  8. Piperlongumine selectively suppresses ABC-DLBCL through inhibition of NF-κB p65 subunit nuclear import.

    Niu, Mingshan; Shen, Yangling; Xu, Xiaoyu; Yao, Yao; Fu, Chunling; Yan, Zhiling; Wu, Qingyun; Cao, Jiang; Sang, Wei; Zeng, Lingyu; Li, Zhenyu; Liu, Xuejiao; Xu, Kailin

    2015-07-10

    Constitutive NF-κB activation is required for survival of activated B cell-like subtype of diffuse large B cell lymphoma (ABC-DLBCL). However, current NF-κB targeting strategies lack cancer cell specificity. Here, we identified a novel inhibitor, piperlongumine, features direct binding to NF-κB p65 subunit and suppression of p65 nuclear import. This was accompanied by NF-κB reporter activity suppression and NF-κB target gene downregulation. Moreover, mutation of Cys(38) to Ser in p65 abolished this effect of piperlongumine on inhibition of p65 nuclear import. Furthermore, we show that piperlongumine selectively inhibited proliferation and induced apoptosis of ABC-DLBCL cells. Most notably, it has been reported that piperlongumine did not affect normal cells even at high doses and was nontoxic to animals. Hence, our current study provides new insight into piperlongumine's mechanism of action and novel approach to ABC-DLBCL target therapy. PMID:25979358

  9. ABC3 Part I: a review of the guidelines for terminology, classification and management of cervical cytology in England.

    Smith, J H F

    2012-12-01

    The provision of guidance on cytology reporting and evaluation, first outlined in 1995 with the publication of Achievable Standards, Benchmarks for Reporting, and Criteria for Evaluating Cervical Cytopathology (ABC), and subsequently revised and expanded in a second edition in 2000, has been pivotal to the success of the National Health Service Cervical Screening Programme (NHSCSP), ensuring that standards are upheld, and that rigorous evaluation and quality assurance take place. In the last decade, major changes to the NHSCSP, notably the adoption of revised age ranges and screening intervals for all women in England, implementation of liquid-based cytology and, most recently, the decision to introduce high-risk human papillomavirus (HR-HPV) testing for triage of low-grade and borderline (equivalent to 'atypical') cytological abnormalities and test of cure after treatment of cervical intraepithelial neoplasia (CIN) determined that an updated version of ABC was required. The third edition of ABC recommends adoption, with minor modification, of the revised British Society for Clinical Cytology terminology and provides guidance on the management of abnormal cytology results linked to this terminology taking account of HR-HPV testing. To accommodate these changes, expanded result codes, which are electronic codes used to transfer management information to central computers for follow-up, call and recall of individual women, have been developed. Further guidance on specimen adequacy is also provided. Revised performance indicators are described and explained in a separate article by R. Blanks in this issue of Cytopathology. All the changes in ABC3 are designed to support the mission statement of the NHSCSP that 'the objective of cervical screening is to reduce cervical cancer incidence and mortality by screening with a high sensitivity for the detection of CIN2 or worse, whilst maintaining a high specificity'. PMID:23173804

  10. The Boron Efflux Transporter ROTTEN EAR Is Required for Maize Inflorescence Development and Fertility[C][W][OPEN

    Chatterjee, Mithu; Tabi, Zara; Galli, Mary; Malcomber, Simon; Buck, Amy; Muszynski, Michael; Gallavotti, Andrea

    2014-01-01

    Although boron has a relatively low natural abundance, it is an essential plant micronutrient. Boron deficiencies cause major crop losses in several areas of the world, affecting reproduction and yield in diverse plant species. Despite the importance of boron in crop productivity, surprisingly little is known about its effects on developing reproductive organs. We isolated a maize (Zea mays) mutant, called rotten ear (rte), that shows distinct defects in vegetative and reproductive development, eventually causing widespread sterility in its inflorescences, the tassel and the ear. Positional cloning revealed that rte encodes a membrane-localized boron efflux transporter, co-orthologous to the Arabidopsis thaliana BOR1 protein. Depending on the availability of boron in the soil, rte plants show a wide range of phenotypic defects that can be fully rescued by supplementing the soil with exogenous boric acid, indicating that rte is crucial for boron transport into aerial tissues. rte is expressed in cells surrounding the xylem in both vegetative and reproductive tissues and is required for meristem activity and organ development. We show that low boron supply to the inflorescences results in widespread defects in cell and cell wall integrity, highlighting the structural importance of boron in the formation of fully fertile reproductive organs. PMID:25035400

  11. A nucleotide sugar transporter involved in glycosylation of the Toxoplasma tissue cyst wall is required for efficient persistence of bradyzoites.

    Carolina E Caffaro

    Full Text Available Toxoplasma gondii is an intracellular parasite that transitions from acute infection to a chronic infective state in its intermediate host via encystation, which enables the parasite to evade immune detection and clearance. It is widely accepted that the tissue cyst perimeter is highly and specifically decorated with glycan modifications; however, the role of these modifications in the establishment and persistence of chronic infection has not been investigated. Here we identify and biochemically and biologically characterize a Toxoplasma nucleotide-sugar transporter (TgNST1 that is required for cyst wall glycosylation. Toxoplasma strains deleted for the TgNST1 gene (Δnst1 form cyst-like structures in vitro but no longer interact with lectins, suggesting that Δnst1 strains are deficient in the transport and use of sugars for the biosynthesis of cyst-wall structures. In vivo infection experiments demonstrate that the lack of TgNST1 activity does not detectably impact the acute (tachyzoite stages of an infection or tropism of the parasite for the brain but that Δnst1 parasites are severely defective in persistence during the chronic stages of the infection. These results demonstrate for the first time the critical role of parasite glycoconjugates in the persistence of Toxoplasma tissue cysts.

  12. A component of the Sec61 ER protein transporting pore is required for plant susceptibility to powdery mildew

    Zhang, Wen-Jing; Hanisch, Susanne; Kwaaitaal, Mark Adrianus Cornelis J;

    2013-01-01

    EHM, the fungus obtains nutrients from and secretes effector proteins into the plant cell. In the plant cell these effectors interfere with cellular processes such as pathogen defense and membrane trafficking. However, the mechanisms behind effector delivery are largely unknown. This paper provides a...... retrotranslocon pores, the latter being part of the ER-associated protein degradation machinery. We provide support for a model suggesting that the retrotranslocon function of HvSec61βa is required for successful powdery mildew fungal infection. HvSec61βa-GFP and a luminal ER marker were co-localized to the ER....... Effector transport across this EHM-ER interface may occur by a vesicle-mediated process, while the Sec61 retrotranslocon pore potentially provides an escape route for these proteins to reach the cytosol....

  13. Requirements for design of accelerator, beam transport, and target in a study of thermonuclear reaction cross section

    Itahashi, T.; Takahisa, K.; Fujiwara, M.; Toki, H.; Ejiri, H. [Osaka Univ., Ibaraki (Japan). Research Center for Nuclear Physics; Ohsumi, H.; Komori, M.

    1997-03-01

    A compact accelerator with high current ion source, low energy beam transport elements and windowless gas target was designed to investigate the thermonuclear reaction cross section. The idea of this project focused on the cross section measurement of the fusion reaction data {sup 3}He+{sup 3}He-{sup 4}He+p+p at 25keV. The system will be installed in Otoh Cosmo Observatory (1270m.w.e.) to get rid of the huge cosmic and environmental background. It consists of NANOGUN ECR ion source, focusing elements made of permanent magnets window less {sup 3}He gas target and/or He{sup 3} plasma target and detector telescopes with low noise and low background. Requirements for these were discussed technically and various ideas were proposed. (author)

  14. Object Detection Based on Template Matching through Use of Best-So-Far ABC

    Anan Banharnsakun

    2014-01-01

    Full Text Available Best-so-far ABC is a modified version of the artificial bee colony (ABC algorithm used for optimization tasks. This algorithm is one of the swarm intelligence (SI algorithms proposed in recent literature, in which the results demonstrated that the best-so-far ABC can produce higher quality solutions with faster convergence than either the ordinary ABC or the current state-of-the-art ABC-based algorithm. In this work, we aim to apply the best-so-far ABC-based approach for object detection based on template matching by using the difference between the RGB level histograms corresponding to the target object and the template object as the objective function. Results confirm that the proposed method was successful in both detecting objects and optimizing the time used to reach the solution.

  15. Object Detection Based on Template Matching through Use of Best-So-Far ABC

    2014-01-01

    Best-so-far ABC is a modified version of the artificial bee colony (ABC) algorithm used for optimization tasks. This algorithm is one of the swarm intelligence (SI) algorithms proposed in recent literature, in which the results demonstrated that the best-so-far ABC can produce higher quality solutions with faster convergence than either the ordinary ABC or the current state-of-the-art ABC-based algorithm. In this work, we aim to apply the best-so-far ABC-based approach for object detection based on template matching by using the difference between the RGB level histograms corresponding to the target object and the template object as the objective function. Results confirm that the proposed method was successful in both detecting objects and optimizing the time used to reach the solution. PMID:24812556

  16. Object detection based on template matching through use of best-so-far ABC.

    Banharnsakun, Anan; Tanathong, Supannee

    2014-01-01

    Best-so-far ABC is a modified version of the artificial bee colony (ABC) algorithm used for optimization tasks. This algorithm is one of the swarm intelligence (SI) algorithms proposed in recent literature, in which the results demonstrated that the best-so-far ABC can produce higher quality solutions with faster convergence than either the ordinary ABC or the current state-of-the-art ABC-based algorithm. In this work, we aim to apply the best-so-far ABC-based approach for object detection based on template matching by using the difference between the RGB level histograms corresponding to the target object and the template object as the objective function. Results confirm that the proposed method was successful in both detecting objects and optimizing the time used to reach the solution. PMID:24812556

  17. Robust LogitBoost and Adaptive Base Class (ABC) LogitBoost

    Li, Ping

    2012-01-01

    Logitboost is an influential boosting algorithm for classification. In this paper, we develop robust logitboost to provide an explicit formulation of tree-split criterion for building weak learners (regression trees) for logitboost. This formulation leads to a numerically stable implementation of logitboost. We then propose abc-logitboost for multi-class classification, by combining robust logitboost with the prior work of abc-boost. Previously, abc-boost was implemented as abc-mart using the mart algorithm. Our extensive experiments on multi-class classification compare four algorithms: mart, abcmart, (robust) logitboost, and abc-logitboost, and demonstrate the superiority of abc-logitboost. Comparisons with other learning methods including SVM and deep learning are also available through prior publications.

  18. Chondroitinase ABC plus bone marrow mesenchymal stem cells for repair of spinal cord injury☆

    Zhang, Chun; He, Xijing; Li, Haopeng; Wang, Guoyu

    2013-01-01

    As chondroitinase ABC can improve the hostile microenvironment and cell transplantation is proven to be effective after spinal cord injury, we hypothesized that their combination would be a more effective treatment option. At 5 days after T8 spinal cord crush injury, rats were injected with bone marrow mesenchymal stem cell suspension or chondroitinase ABC 1 mm from the edge of spinal cord damage zone. Chondroitinase ABC was first injected, and bone marrow mesenchymal stem cell suspension was...

  19. A Common Weight Linear Optimization Approach for Multicriteria ABC Inventory Classification

    S. M. Hatefi; Torabi, S. A.

    2015-01-01

    Organizations typically employ the ABC inventory classification technique to have an efficient control on a huge amount of inventory items. The ABC inventory classification problem is classification of a large amount of items into three groups: A, very important; B, moderately important; and C, relatively unimportant. The traditional ABC classification only accounts for one criterion, namely, the annual dollar usage of the items. But, there are other important criteria in real world which str...

  20. Synthesis and development of ordered, phase-separated, room-temperature ionic liquid-based AB and ABC block copolymers for gas separation applications

    Wiesenauer, Erin F.

    CO2 capture process development is an economically and environmentally important challenge, as concerns over greenhouse gas emissions continue to receive worldwide attention. Many applications require the separation of CO 2 from other light gases such as N2, CH4, and H2 and a number of technologies have been developed to perform such separations. While current membrane technology offers an economical, easy to operate and scale-up solution, polymeric membranes cannot withstand high temperatures and aggressive chemical environments, and they often exhibit an unfavorable tradeoff between permeability and selectivity. Room-temperature ionic-liquids (RTILs) are very attractive as next-generation CO2-selective separation media and their development into polymerized membranes combat these challenges. Furthermore, polymers that can self-assemble into nanostructured, phase-separated morphologies (e.g., block copolymers, BCPs) have a direct effect on gas transport as materials morphology can influence molecular diffusion and membrane transport performance. In this thesis, nanophase-separated, RTIL-based AB and ABC di- and tri-BCPs were prepared via the sequential, living ring-opening metathesis polymerization (ROMP) of an IL-based monomer and one or more mutually immiscible co-monomers. This novel type of ion-containing BCP system forms various ordered nanostructures in the melt state via primary and secondary structure control. Monomer design and control of block composition, sequence, and overall polymer lengths were found to directly affect the ordered polymer assembly. Supported, composite membranes of these new BCPs were successfully fabricated, and the effect of BCP composition and nanostructure on CO2/light gas transport properties was studied. These nanostructured IL-based BCPs represent innovative polymer architectures and show great potential CO2/light gas membrane separation applications.

  1. Multidrug transport by ATP binding cassette transporters : a proposed two-cylinder engine mechanism

    van Veen, HW; Higgins, CF; Konings, WN

    2001-01-01

    The elevated expression of ATP binding cassette (ABC) multidrug transporters in multidrug-resistant cells interferes with the drug-based control of cancers and infectious pathogenic microorganisms. Multidrug transporters interact directly with the drug substrates. This review summarizes current insi

  2. IMPROVED SEARCH MECHANISM IN ABC AND ITS APPLICATION IN ENGINEERING DESIGN PROBLEMS

    TARUN KUMAR SHARMA

    2015-01-01

    Full Text Available ABC (Artificial Bee Colony is one of the most recent nature inspired algorithm (NIA based on swarming metaphor. Proposed by Karaboga in 2005, ABC has proven to be a robust and efficient algorithm for solving global optimization problems over continuous space. In this paper, we propose a modified version of the ABC. In modified version called Dichotomous ABC (DABC, the idea is to move dichotomously in both directions to generate a new trial point. The results of a trade study carried out on three classical structural optimization problems taken from literature indicate the validity of the proposed algorithm.

  3. Developing a utility index for the Aberrant Behavior Checklist (ABC-C) for fragile X syndrome

    Kerr, Cicely; Breheny, Katie; Lloyd, Andrew; Brazier, John; Bailey, Donald B.; Berry-Kravis, Elizabeth; Cohen, Jonathan; Petrillo, Jennifer

    2014-01-01

    Purpose This study aimed to develop a utility index (the ABC-UI) from the Aberrant Behavior Checklist-Community (ABC-C), for use in quantifying the benefit of emerging treatments for fragile X syndrome (FXS). Methods The ABC-C is a proxy-completed assessment of behaviour and is a widely used measure in FXS. A subset of ABC-C items across seven dimensions was identified to include in health state descriptions. This item reduction process was based on item performance, factor analysis and Rasch...

  4. Transport: Introduction

    Lewis, William; Rosenberg, Sanders D.

    1992-01-01

    Space transportation requirements for the NASA baseline scenario for future space missions are discussed. Spacecraft/propulsion technologies required for surface-to-orbit, orbit-to-orbit, and surface (lunar) transportation are addressed.

  5. Characterization of a novel domain ‘GATE’ in the ABC protein DrrA and its role in drug efflux by the DrrAB complex

    Zhang, Han; Rahman, Sadia; Li, Wen; Fu, Guoxing; Kaur, Parjit, E-mail: pkaur@gsu.edu

    2015-03-27

    A novel domain, GATE (Glycine-loop And Transducer Element), is identified in the ABC protein DrrA. This domain shows sequence and structural conservation among close homologs of DrrA as well as distantly-related ABC proteins. Among the highly conserved residues in this domain are three glycines, G215, G221 and G231, of which G215 was found to be critical for stable expression of the DrrAB complex. Other conserved residues, including E201, G221, K227 and G231, were found to be critical for the catalytic and transport functions of the DrrAB transporter. Structural analysis of both the previously published crystal structure of the DrrA homolog MalK and the modeled structure of DrrA showed that G215 makes close contacts with residues in and around the Walker A motif, suggesting that these interactions may be critical for maintaining the integrity of the ATP binding pocket as well as the complex. It is also shown that G215A or K227R mutation diminishes some of the atomic interactions essential for ATP catalysis and overall transport function. Therefore, based on both the biochemical and structural analyses, it is proposed that the GATE domain, located outside of the previously identified ATP binding and hydrolysis motifs, is an additional element involved in ATP catalysis. - Highlights: • A novel domain ‘GATE’ is identified in the ABC protein DrrA. • GATE shows high sequence and structural conservation among diverse ABC proteins. • GATE is located outside of the previously studied ATP binding and hydrolysis motifs. • Conserved GATE residues are critical for stability of DrrAB and for ATP catalysis.

  6. Characterization of a novel domain ‘GATE’ in the ABC protein DrrA and its role in drug efflux by the DrrAB complex

    A novel domain, GATE (Glycine-loop And Transducer Element), is identified in the ABC protein DrrA. This domain shows sequence and structural conservation among close homologs of DrrA as well as distantly-related ABC proteins. Among the highly conserved residues in this domain are three glycines, G215, G221 and G231, of which G215 was found to be critical for stable expression of the DrrAB complex. Other conserved residues, including E201, G221, K227 and G231, were found to be critical for the catalytic and transport functions of the DrrAB transporter. Structural analysis of both the previously published crystal structure of the DrrA homolog MalK and the modeled structure of DrrA showed that G215 makes close contacts with residues in and around the Walker A motif, suggesting that these interactions may be critical for maintaining the integrity of the ATP binding pocket as well as the complex. It is also shown that G215A or K227R mutation diminishes some of the atomic interactions essential for ATP catalysis and overall transport function. Therefore, based on both the biochemical and structural analyses, it is proposed that the GATE domain, located outside of the previously identified ATP binding and hydrolysis motifs, is an additional element involved in ATP catalysis. - Highlights: • A novel domain ‘GATE’ is identified in the ABC protein DrrA. • GATE shows high sequence and structural conservation among diverse ABC proteins. • GATE is located outside of the previously studied ATP binding and hydrolysis motifs. • Conserved GATE residues are critical for stability of DrrAB and for ATP catalysis

  7. Modern requirements of legal regulations for transport in the cities and necessity for making sector strategy in order to harmonize with other modes of transport

    Aleksandra VASILJ

    2010-01-01

    Full Text Available Crucial problem of transport in the cities, as a source of pollution of the environment, are uncontrolled individual motorization, permanently increasing number of motor vehicles on the roads and streets that expels human primary need for walking.Reduced social area is also loaded with vibrations and increased sound pressure which often exceeds recommended 80 dB/A.Accent is on ecological problems caused by: polluted air, which comes from gas emissions (full of aerosol, metals, dust, soot, smoke, cutting and disappearance of green areas in order to provide space for transport, making huge amount of secondary waste (e. g. tyres, metal, used oil, liquids.To improve present sequences and reduce negative effects in the future it is necessary to take different measures: fiscal, administrative, educational, which will restrict use of individual, particularly car traffic. Same measures should be used to encourage users to use public transportation.Transport in the cities, with all weaknesses and comparative advantages, should be separately analyzed mode of road transport and also be an individual part in strategic documents.The most important questions that such strategy of transport in the cities should include are: regulation, privatization or other more efficient mode of organization in public transport of passengers, as well as urban elements of sustainable development of the urban areas and transport.

  8. NasFED proteins mediate assimilatory nitrate and nitrite transport in Klebsiella oxytoca (pneumoniae) M5al.

    Wu, Q; Stewart, V

    1998-03-01

    Klebsiella oxytoca can use nitrate and nitrite as sole nitrogen sources. The enzymes required for nitrate and nitrite assimilation are encoded by the nasFEDCBA operon. We report here the complete nasFED sequence. Sequence comparisons indicate that the nasFED genes encode components of a conventional periplasmic binding protein-dependent transport system consisting of a periplasmic binding protein (NasF), a homodimeric intrinsic membrane protein (NasE), and a homodimeric ATP-binding cassette (ABC) protein (NasD). The NasF protein and the related NrtA and CmpA proteins of cyanobacteria contain leader (signal) sequences with the double-arginine motif that is hypothesized to direct prefolded proteins to an alternate protein export pathway. The NasE protein and the related NrtB and CmpB proteins of cyanobacteria contain unusual variants of the EAA loop sequence that defines membrane-intrinsic proteins of ABC transporters. To characterize nitrate and nitrite transport, we constructed in-frame nonpolar deletions of the chromosomal nasFED genes. Growth tests coupled with nitrate and nitrite uptake assays revealed that the nasFED genes are essential for nitrate transport and participate in nitrite transport as well. Interestingly, the delta nasF strain exhibited leaky phenotypes, particularly at elevated nitrate concentrations, suggesting that the NasED proteins are not fully dependent on the NasF protein. PMID:9495773

  9. Desenvolvimento Regional Sustentável no Grande ABC: análise das contribuições de uma instituição financeira brasileira

    Nelson Correa Oliveira Junior

    2011-01-01

    Full Text Available This work provided a reflection on the contributions of regional sustainable development plans (DRS of Banco do Brasil (BB to the region known as Grande ABC through the analysis of the plans offered by the institution in documents and website, as well as by interviews conducted with the plan's stakeholders. A bibliographic research has been made, where the concepts of development, regional development, sustainable development and sustainable regional development are discussed and detailed. Then, a document research has been made, which allowed the verifying of the data of the plans in the country and in the region of Grande ABC, also providing subsidies to the definition of the stakeholders interviewed in the survey. The methodology used was the exploratory research which focuses on better understanding and familiarization of the researcher and the matter being considered. It was found that the DRS plans at the Grande ABC still require new partnerships and actions that can boost their development, the need for bigger monitoring, control and dissemination of the concepts of DRS by Banco do Brasil; the concern in establishing strategic partnerships has been noticed, but still without the participation of the academy in a more constant way, finally, it is understood that the Regional Sustainable Development of Banco do Brasil at Grande ABC, according to their social, environmental, cultural and economic contributions, can not be considered an initiative that has significant impacts yet, but it is a way to search for funds and partnerships for the formulation of local and regional development initiatives.

  10. Intragenic Suppressing Mutations Correct the Folding and Intracellular Traffic of Misfolded Mutants of Yor1p, a Eukaryotic Drug Transporter*

    Pagant, Silvere; Halliday, John J.; Kougentakis, Christos; Miller, Elizabeth A.

    2010-01-01

    ATP-binding cassette (ABC) transporters play pivotal physiological roles in substrate transport across membranes, and defective assembly of these proteins can cause severe disease associated with improper drug or ion flux. The yeast protein Yor1p is a useful model to study the biogenesis of ABC transporters; deletion of a phenylalanine residue in the first nucleotide-binding domain (NBD1) causes misassembly and retention in the endoplasmic reticulum (ER) of the resulting protein Yor1p-ΔF670, ...

  11. Low levels of graphene and graphene oxide inhibit cellular xenobiotic defense system mediated by efflux transporters.

    Liu, Su; Jiang, Wei; Wu, Bing; Yu, Jing; Yu, Haiyan; Zhang, Xu-Xiang; Torres-Duarte, Cristina; Cherr, Gary N

    2016-06-01

    Low levels of graphene and graphene oxide (GO) are considered to be environmentally safe. In this study, we analyzed the potential effects of graphene and GO at relatively low concentrations on cellular xenobiotic defense system mediated by efflux transporters. The results showed that graphene (<0.5 μg/mL) and GO (<20 μg/mL) did not decrease cell viability, generate reactive oxygen species, or disrupt mitochondrial function. However, graphene and GO at the nontoxic concentrations could increase calcein-AM (CAM, an indicator of membrane ATP-binding cassette (ABC) transporter) activity) accumulation, indicating inhibition of ABC transporters' efflux capabilities. This inhibition was observed even at 0.005 μg/mL graphene and 0.05 μg/mL GO, which are 100 times and 400 times lower than their lowest toxic concentration from cytotoxicity experiments, respectively. The inhibition of ABC transporters significantly increased the toxicity of paraquat and arsenic, known substrates of ABC transporters. The inhibition of ABC transporters was found to be based on graphene and GO damaging the plasma membrane structure and fluidity, thus altering functions of transmembrane ABC transporters. This study demonstrates that low levels of graphene and GO are not environmentally safe since they can significantly make cell more susceptible to other xenobiotics, and this chemosensitizing activity should be considered in the risk assessment of graphene and GO. PMID:26554512

  12. Redundancy Level Optimization in Modular Software System Models using ABC

    Tarun Kumar Sharma; Millie Pant

    2014-01-01

    The performance of optimization algorithms is problem dependent and as per no free lunch theorem, there exists no such algorithm which can be efficiently applied to every type of problem(s). However, we can modify the algorithm/ technique in a manner such that it is able to deal with a maximum type of problems. In this study we have modified the structure of basic Artificial Bee Colony (ABC), a recently proposed metaheuristic algorithm based on the concept of swarm intelligence to optimize t...

  13. ABA, AAB and ABC renewal in taste aversion learning

    Javier Nieto

    2012-01-01

    Full Text Available Existe evidencia de que los cambios en los estímulos ambientales afectan activamente la ocurrencia de la respuesta condicionada (RC. La renovación contextual o la reaparición de una RC extinguida se observa cuando se continúa la extinción de dicha respuesta en un contexto diferente. Se diseñaron dos experimentos con el propósito de evaluar la renovación contextual en un procedimiento de condicionamiento de aversión al sabor. En el Experimento 1, se analizó la renovación contextual ABA y AAB, mientras que el Experimento 2 se evaluó la renovación ABA, AAB y ABC. Ambos experimentos constaron de tres fases: adquisición, extinción y prueba. En ambos experimentos, la fase de adquisición se condujo en el Contexto A para todos los grupos y en ella, se inyectó i.p. Cloruro de Litio (LiCl a los sujetos inmediatamente después de que ingirieron una solución de sacarosa. En la fase de extinción, se expuso a los sujetos únicamente a la solución de sacarosa, sin la inyección. Los grupos AAB y AAA recibieron esta fase en el Contexto A, mientras que los sujetos de los grupos ABA y ABC la recibieron en el Contexto B. En la fase de prueba, se continúo la extinción. Los grupos ABA y AAA recibieron esta fase en el Contexto A, el grupo AAB en el contexto B, mientras el grupo ABC la recibió en un tercer contexto. Los resultados mostraron renovación de la aversión condicionada al sabor en los grupos ABA, AAB y ABC. Se analiza la relación de estos hallazgos con el modelo de recuperación de información de Bouton (1994.

  14. Design of the storage location based on the ABC analyses

    Jemelka, Milan; Chramcov, Bronislav; Kříž, Pavel

    2016-06-01

    The paper focuses on process efficiency and saving storage costs. Maintaining inventory through putaway strategy takes personnel time and costs money. The aim is to control inventory in the best way. The ABC classification based on Villefredo Pareto theory is used for a design of warehouse layout. New design of storage location reduces the distance of fork-lifters, total costs and it increases inventory process efficiency. The suggested solutions and evaluation of achieved results are described in detail. Proposed solutions were realized in real warehouse operation.

  15. 24 CFR 3280.903 - General requirements for designing the structure to withstand transportation shock and vibration.

    2010-04-01

    ... the structure to withstand transportation shock and vibration. 3280.903 Section 3280.903 Housing and... structure to withstand transportation shock and vibration. (a) The cumulative effect of highway transportation shock and vibration upon a manufactured home structure may result in incremental degradation...

  16. A C-terminal PDZ domain-binding sequence is required for striatal distribution of the dopamine transporter

    Rickhag, Karl Mattias; Hansen, Freja Herborg; Sørensen, Gunnar;

    2013-01-01

    The dopamine transporter mediates reuptake of dopamine from the synaptic cleft. The cellular mechanisms controlling dopamine transporter levels in striatal nerve terminals remain poorly understood. The dopamine transporters contain a C-terminal PDZ (PSD-95/Discs-large/ZO-1) domain-binding sequenc...

  17. Rice Stomatal Closure Requires Guard Cell Plasma Membrane ATP-Binding Cassette Transporter RCN1/OsABCG5.

    Matsuda, Shuichi; Takano, Sho; Sato, Moeko; Furukawa, Kaoru; Nagasawa, Hidetaka; Yoshikawa, Shoko; Kasuga, Jun; Tokuji, Yoshihiko; Yazaki, Kazufumi; Nakazono, Mikio; Takamure, Itsuro; Kato, Kiyoaki

    2016-03-01

    Water stress is one of the major environmental stresses that affect agricultural production worldwide. Water loss from plants occurs primarily through stomatal pores. Here, we report that an Oryza sativa half-size ATP-binding cassette (ABC) subfamily G protein, RCN1/OsABCG5, is involved in stomatal closure mediated by phytohormone abscisic acid (ABA) accumulation in guard cells. We found that the GFP-RCN1/OsABCG5-fusion protein was localized at the plasma membrane in guard cells. The percentage of guard cell pairs containing both ABA and GFP-RCN1/OsABCG5 increased after exogenous ABA treatment, whereas they were co-localized in guard cell pairs regardless of whether exogenous ABA was applied. ABA application resulted in a smaller increase in the percentage of guard cell pairs containing ABA in rcn1 mutant (A684P) and RCN1-RNAi than in wild-type plants. Furthermore, polyethylene glycol (drought stress)-inducible ABA accumulation in guard cells did not occur in rcn1 mutants. Stomata closure mediated by exogenous ABA application was strongly reduced in rcn1 mutants. Finally, rcn1 mutant plants had more rapid water loss from detached leaves than the wild-type plants. These results indicate that in response to drought stress, RCN1/OsABCG5 is involved in accumulation of ABA in guard cells, which is indispensable for stomatal closure. PMID:26708605

  18. ATP binding cassette transporter gene expression in rat liver progenitor cells

    2003-01-01

    Background and aim: Liver regeneration after severe liver damage depends in part on proliferation and differentiation of hepatic progenitor cells (HPCs). Under these conditions they must be able to withstand the toxic milieu of the damaged liver. ATP binding cassette (ABC) transporters are cytoprotective efflux pumps that may contribute to the preservation of these cells. The aim of this study was to determine the ABC transporter phenotype of HPCs.

  19. Not used to public transport any longer. A study about transport service based on needs and requirements in Siljansnaes; Man har vant sig av med kollektivtrafik. En studie om transportservice baserad paa behov och vanor i Siljansnaes

    Warsen, Lisa

    2000-05-01

    The goal of the transport policy is that every citizen shall have a transport service on a reasonable level. In sparsely populated areas the public transport service is on a low level and many groups without a private car will have small possibilities to travel to different activities. Siljansnaes, with nearly 2000 citizens, is a small region in the community Leksand in Dalarna where the transport problem have been more tangible. The aim of this project has been to create a transport system more based on the needs and requirements. Also the people who lives in Siljansnaes in their leisure times are a part in the project. They are almost as many as the permanent living ones. The result also has been built on the goal of a higher care of the environment. The transport solution contains; village-bus, car-pooling, changed price system for public transport, but also a partnership between the citizens, the community and the regional transport company (Dalatrafik). The project as a process has the goal that the people in Siljansnaes should take part of the solution and the responsibility for the operation of the system.

  20. A Quantitative bgl Operon Model for E. coli Requires BglF Conformational Change for Sugar Transport

    Chopra, Paras; Bender, Andreas

    The bgl operon is responsible for the metabolism of β-glucoside sugars such as salicin or arbutin in E. coli. Its regulatory system involves both positive and negative feedback mechanisms and it can be assumed to be more complex than that of the more closely studied lac and trp operons. We have developed a quantitative model for the regulation of the bgl operon which is subject to in silico experiments investigating its behavior under different hypothetical conditions. Upon administration of 5mM salicin as an inducer our model shows 80-fold induction, which compares well with the 60-fold induction measured experimentally. Under practical conditions 5-10mM inducer are employed, which is in line with the minimum inducer concentration of 1mM required by our model. The necessity of BglF conformational change for sugar transport has been hypothesized previously, and in line with those hypotheses our model shows only minor induction if conformational change is not allowed. Overall, this first quantitative model for the bgl operon gives reasonable predictions that are close to experimental results (where measured). It will be further refined as values of the parameters are determined experimentally. The model was developed in Systems Biology Markup Language (SBML) and it is available from the authors and from the Biomodels repository [www.ebi.ac.uk/biomodels].

  1. Creating an iPhone Application for Collecting Continuous ABC Data

    Whiting, Seth W.; Dixon, Mark R.

    2012-01-01

    This paper provides an overview and task analysis for creating a continuous ABC data- collection application using Xcode on a Mac computer. Behavior analysts can program an ABC data collection system, complete with a customized list of target clients, antecedents, behaviors, and consequences to be recorded, and have the data automatically sent to…

  2. Chondroitinase ABC treatment of injured spinal cord in rats Evaluation of long-term outcomes

    Haifeng Yuan; Qingquan Kong; Yongli Ding; Yueming Song; Lihong Hu; Zili Wang; Hao Liu; Limin Liu; Quan Gong; Tao Li

    2010-01-01

    Chondroitin sulfate proteoglycans(CSPGs)which are produced by mature oligodendrocytes and reactive astrocytes can be upregulated after spinal cord injury and contribute to regenerative failure.Chondroitinase ABC(ChABC)digests glycosaminoglycan chains on CSPGs and can thereby overcome CSPG-mediated inhibition.However,many current studies have used an incomplete spinal cord injury model,and examined results after 8 12 weeks of ChABC treatment.In this study,a complete rat spinal cord transection injury model was used to study the long-term effects of ChABC treatment by subarachnoid catheter.Pathology of spinal cord regeneration was compared with control 24 weeks following ChABC treatment using immunohistochemistry and axon tracing techniques.At 24 weeks after injury,neurofilament 200 expression was significantly greater in the ChABC treatment group compared with the transection group.In the ChABC treatment group,axonal growth was demonstrated by a large number of biotinylated dextran amine positive axons caudal to,or past,the epicenter of injury.Biotinylated dextran amine-labeled fibers were found in the proximal end of the spinal cord in the transection alone group.These results confirm that ChABC can promote axon growth,neural regeneration,and repair after spinal cord injury in rats long after the initial injury.

  3. ABCE1 is essential for S phase progression in human cells.

    Toompuu, Marina; Kärblane, Kairi; Pata, Pille; Truve, Erkki; Sarmiento, Cecilia

    2016-05-01

    ABCE1 is a highly conserved protein universally present in eukaryotes and archaea, which is crucial for the viability of different organisms. First identified as RNase L inhibitor, ABCE1 is currently recognized as an essential translation factor involved in several stages of eukaryotic translation and ribosome biogenesis. The nature of vital functions of ABCE1, however, remains unexplained. Here, we study the role of ABCE1 in human cell proliferation and its possible connection to translation. We show that ABCE1 depletion by siRNA results in a decreased rate of cell growth due to accumulation of cells in S phase, which is accompanied by inefficient DNA synthesis and reduced histone mRNA and protein levels. We infer that in addition to the role in general translation, ABCE1 is involved in histone biosynthesis and DNA replication and therefore is essential for normal S phase progression. In addition, we analyze whether ABCE1 is implicated in transcript-specific translation via its association with the eIF3 complex subunits known to control the synthesis of cell proliferation-related proteins. The expression levels of a few such targets regulated by eIF3A, however, were not consistently affected by ABCE1 depletion. PMID:26985706

  4. The ABCs of Activity-Based Costing: A Cost Containment and Reallocation Tool.

    Turk, Frederick J.

    1992-01-01

    This article describes activity-based costing (ABC) and how this tool may help management understand the costs of major activities and identify possible alternatives. Also discussed are the traditional costing systems used by higher education and ways of applying ABC to higher education. (GLR)

  5. The Role of Activity Based Costing (ABC) in Educational Support Services: A White Paper.

    Edds, Daniel B.

    Many front-line managers who are assuming more financial responsibility for their organizations find traditional cost accounting inadequate for their needs and are turning to Activity Based Costing (ABC). ABC is not a financial reporting system to serve the needs of regulatory agencies, but a tool that tracks costs from the general ledger…

  6. The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.

    Jin Hwan Park

    2015-09-01

    Full Text Available A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose.

  7. 41 CFR 102-118.25 - Does GSA still require my agency to submit its overall transportation policies for approval?

    2010-07-01

    ... Public Contracts and Property Management Federal Property Management Regulations System (Continued) FEDERAL MANAGEMENT REGULATION TRANSPORTATION 118-TRANSPORTATION PAYMENT AND AUDIT General Introduction... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false Does GSA still...

  8. Analysis of transport logistics and routing requirements for radioactive waste management systems with respect to a minimum power scenario

    This report assesses the transport logistics associated with disposal of intermediate-level radioactive waste, as generated by CEGB, SSEB, UKAEA and BNFL, in accordance with a 'Minimum Power Scenario'. Transport by road and rail is analysed, as in previous reports; use of coastal shipping however has not been included but has been replaced with a combined road/rail option. (author)

  9. Plant pleiotropic drug resistance transporters:Transport mechanism, gene expression, and function

    Mohammed Nuruzzaman; Ru Zhang; Hong-Zhe Cao; Zhi-Yong Luo

    2014-01-01

    Pleiotropic drug resistance (PDR) transporters belonging to the ABCG subfamily of ATP-binding cassette (ABC) transporters are identified only in fungi and plants. Members of this family are expressed in plants in response to various biotic and abiotic stresses and transport a diverse array of molecules across membranes. Although their detailed transport mechanism is largely unknown, they play important roles in detoxification processes, preventing water loss, transport of phytohormones, and secondary metabolites. This review provides insights into transport mechanisms of plant PDR transporters, their expression profiles, and multitude functions in plants.

  10. Docking of the Periplasmic FecB Binding Protein to the FecCD Transmembrane Proteins in the Ferric Citrate Transport System of Escherichia coli▿

    Braun, Volkmar; Herrmann, Christina

    2007-01-01

    Citrate-mediated iron transport across the cytoplasmic membrane is catalyzed by an ABC transporter that consists of the periplasmic binding protein FecB, the transmembrane proteins FecC and FecD, and the ATPase FecE. Salt bridges between glutamate residues of the binding protein and arginine residues of the transmembrane proteins are predicted to mediate the positioning of the substrate-loaded binding protein on the transmembrane protein, based on the crystal structures of the ABC transporter...

  11. Evaluation of the role of ATP-binding cassette transporters as a defence mechanism against temephos in populations of Aedes aegypti.

    Lima, Estelita Pereira; Goulart, Marília Oliveira Fonseca; Rolim Neto, Modesto Leite

    2014-11-01

    The role of ATP-binding cassette (ABC) transporters in the efflux of the insecticide, temephos, was assessed in the larvae of Aedes aegypti. Bioassays were conducted using mosquito populations that were either susceptible or resistant to temephos by exposure to insecticide alone or in combination with sublethal doses of the ABC transporter inhibitor, verapamil (30, 35 and 40 μM). The best result in the series was obtained with the addition of verapamil (40 μM), which led to a 2x increase in the toxicity of temephos, suggesting that ABC transporters may be partially involved in conferring resistance to the populations evaluated. PMID:25411004

  12. Evaluation of the role of ATP-binding cassette transporters as a defence mechanism against temephos in populations of Aedes aegypti

    Estelita Pereira Lima

    2014-11-01

    Full Text Available The role of ATP-binding cassette (ABC transporters in the efflux of the insecticide, temephos, was assessed in the larvae of Aedes aegypti. Bioassays were conducted using mosquito populations that were either susceptible or resistant to temephos by exposure to insecticide alone or in combination with sublethal doses of the ABC transporter inhibitor, verapamil (30, 35 and 40 μM. The best result in the series was obtained with the addition of verapamil (40 μM, which led to a 2x increase in the toxicity of temephos, suggesting that ABC transporters may be partially involved in conferring resistance to the populations evaluated.

  13. Supersymmetric Theory of Stochastic ABC Model: A Numerical Study

    Ovchinnikov, Igor V; Ensslin, Torsten A; Wang, Kang L

    2016-01-01

    In this paper, we investigate numerically the stochastic ABC model, a toy model in the theory of astrophysical kinematic dynamos, within the recently proposed supersymmetric theory of stochastics (STS). STS characterises stochastic differential equations (SDEs) by the spectrum of the stochastic evolution operator (SEO) on elements of the exterior algebra or differentials forms over the system's phase space, X. STS can thereby classify SDEs as chaotic or non-chaotic by identifying the phenomenon of stochastic chaos with the spontaneously broken topological supersymmetry that all SDEs possess. We demonstrate the following three properties of the SEO, deduced previously analytically and from physical arguments: the SEO spectra for zeroth and top degree forms never break topological supersymmetry, all SDEs possesses pseudo-time-reversal symmetry, and each de Rahm cohomology class provides one supersymmetric eigenstate. Our results also suggests that the SEO spectra for forms of complementary degrees, i.e., k and ...

  14. [ABC supplies classification: a managment tool of costs in nursing].

    Lourenço, Karina Gomes; Castilho, Valéria

    2006-01-01

    The implementation of costs management systems has been extremely helpful to healthcare area owing to their efficacy in cutting expenditures as well as improving service quality. The ABC classification is an applied strategy to stocktaking and control. The research, which consists of an exploratory/descriptive quantitative analysis, has been carried out in order to identify, in a year time period, the demand for supplies at Universidade de Sao Paulo's Hospital. Of 1938 classified materials, 67 itens had been classified that they correspond to the materials with bigger costs for the hospital. 31.3% of these A-Class supplies catalogued items are the nursing materials, more used for the nursing team. PMID:16915729

  15. Observation of an ABC effect in proton-proton collisions

    Dymov, S; Kacharava, A; Khoukaz, A; Komarov, V; Kulessa, P; Kulikov, A; Kurbatov, V; Macharashvili, G; Merzliakov, S; Mielke, M; Mikirtychiants, S; Nekipelov, M; Nioradze, M; Ohm, H; Rathmann, F; Ströher, H; Tsirkov, D; Uzikov, Yu; Valdau, Yu; Wilkin, C; Yaschenko, S; Zalikhanov, B

    2009-01-01

    The cross section for inclusive multipion production in the pp->ppX reaction was measured at COSY-ANKE at four beam energies, 0.8, 1.1, 1.4, and 2.0 GeV, for low excitation energy in the final pp system, such that the diproton quasi-particle is in the 1S0 state. At the three higher energies the missing mass Mx spectra show a strong enhancement at low Mx, corresponding to an ABC effect that moves steadily to larger values as the energy is increased. Despite the missing-mass structure looking very different at 0.8 GeV, the variation with Mx and beam energy are consistent with two-pion production being mediated through the excitation of two Delta(1232) isobars, coupled to S-- and D-- states of the initial pp system.

  16. Supramolecular Polymers Formed by ABC Miktoarm Star Peptides.

    Lin, Yi-An; Ou, Yu-Chuan; Cheetham, Andrew G; Cui, Honggang

    2013-12-17

    We report here the design and synthesis of an ABC miktoarm star peptide connecting through a lysine junction a short peptide sequence and two hydrophobic but immiscible blocks (a hydrocarbon and a fluorocarbon). The designed molecule can self-assemble into one-dimensional nanostructures with a great diversity of kinetically evolving morphologies in aqueous solution, while molecules that contain only one of the two hydrophobic blocks form structurally similar filaments. We believe the rich assembly behavior and morphological evolution are a direct reflection of the molecular frustration present within the filament core as a result of the in-compatibility of the fluorocarbon and hydrocarbon segments. Our finding opens new opportunities for creating complex supramolecular polymers through the architecture design of small molecular building units. PMID:24490124

  17. Applying Activity Based Costing (ABC Method to Calculate Cost Price in Hospital and Remedy Services

    A Dabiri

    2012-04-01

    Full Text Available Background: Activity Based Costing (ABC is one of the new methods began appearing as a costing methodology in the 1990. It calculates cost price by determining the usage of resources. In this study, ABC method was used for calculating cost price of remedial services in hospitals.Methods: To apply ABC method, Shahid Faghihi Hospital was selected. First, hospital units were divided into three main departments: administrative, diagnostic, and hospitalized. Second, activity centers were defined by the activity analysis method. Third, costs of administrative activity centers were allocated into diagnostic and operational departments based on the cost driver. Finally, with regard to the usage of cost objectives from services of activity centers, the cost price of medical services was calculated.Results: The cost price from ABC method significantly differs from tariff method. In addition, high amount of indirect costs in the hospital indicates that capacities of resources are not used properly.Conclusion: Cost price of remedial services with tariff method is not properly calculated when compared with ABC method. ABC calculates cost price by applying suitable mechanisms but tariff method is based on the fixed price. In addition, ABC represents useful information about the amount and combination of cost price services.

  18. Genome-wide identification and evolution of ATP-binding cassette transporters in the ciliate Tetrahymena thermophila: A case of functional divergence in a multigene family

    Yuan Dongxia

    2010-10-01

    Full Text Available Abstract Background In eukaryotes, ABC transporters that utilize the energy of ATP hydrolysis to expel cellular substrates into the environment are responsible for most of the efflux from cells. Many members of the superfamily of ABC transporters have been linked with resistance to multiple drugs or toxins. Owing to their medical and toxicological importance, members of the ABC superfamily have been studied in several model organisms and warrant examination in newly sequenced genomes. Results A total of 165 ABC transporter genes, constituting a highly expanded superfamily relative to its size in other eukaryotes, were identified in the macronuclear genome of the ciliate Tetrahymena thermophila. Based on ortholog comparisons, phylogenetic topologies and intron characterizations, each highly expanded ABC transporter family of T. thermophila was classified into several distinct groups, and hypotheses about their evolutionary relationships are presented. A comprehensive microarray analysis revealed divergent expression patterns among the members of the ABC transporter superfamily during different states of physiology and development. Many of the relatively recently formed duplicate pairs within individual ABC transporter families exhibit significantly different expression patterns. Further analysis showed that multiple mechanisms have led to functional divergence that is responsible for the preservation of duplicated genes. Conclusion Gene duplications have resulted in an extensive expansion of the superfamily of ABC transporters in the Tetrahymena genome, making it the largest example of its kind reported in any organism to date. Multiple independent duplications and subsequent divergence contributed to the formation of different families of ABC transporter genes. Many of the members within a gene family exhibit different expression patterns. The combination of gene duplication followed by both sequence divergence and acquisition of new patterns of

  19. Report of consultants to the IAEA on requirements for the safe transport of low hazard radioactive materials

    In the present paper changes are recommmended to certain definitions given in the 2nd draft revision of the IAEA 'regulations for the safe transport of radioactive materials (safety series no. 6)'. (orig./RW)

  20. A novel multiprotein complex is required to generate the prechylomicron transport vesicle from intestinal ER[S

    Siddiqi, Shahzad; Saleem, Umair; Abumrad, Nada A.; Davidson, Nicholas O.; Storch, Judith; Siddiqi, Shadab A.; Mansbach, Charles M.

    2010-01-01

    Dietary lipid absorption is dependent on chylomicron production whose rate-limiting step across the intestinal absorptive cell is the exit of chylomicrons from the endoplasmic reticulum (ER) in its ER-to-Golgi transport vesicle, the prechylomicron transport vesicle (PCTV). This study addresses the composition of the budding complex for PCTV. Immunoprecipitation (IP) studies from rat intestinal ER solubilized in Triton X-100 suggested that vesicle-associated membrane protein 7 (VAMP7), apolipo...