Sample records for fab1 fab1 fab1

  1. 75 FR 9438 - Samsung Austin Semiconductor, LLC, DRAM Fab 1, a Subsidiary of Samsung Electronics Corporation... (United States)


    ... Employment and Training Administration Samsung Austin Semiconductor, LLC, DRAM Fab 1, a Subsidiary of Samsung..., applicable to workers of Samsung Austin Semiconductor, LLC, a subsidiary of Samsung Electronics Corporation... Systems, Inc. were employed on-site at the Austin, Texas location of Samsung Austin Semiconductor, LLC,...

  2. Mapping of Fab-1:VEGF Interface Using Carboxyl Group Footprinting Mass Spectrometry (United States)

    Wecksler, Aaron T.; Kalo, Matt S.; Deperalta, Galahad


    A proof-of-concept study was performed to demonstrate that carboxyl group footprinting, a relatively simple, bench-top method, has utility for first-pass analysis to determine epitope regions of therapeutic mAb:antigen complexes. The binding interface of vascular endothelial growth factor (VEGF) and the Fab portion of a neutralizing antibody (Fab-1) was analyzed using carboxyl group footprinting with glycine ethyl ester (GEE) labeling. Tryptic peptides involved in the binding interface between VEGF and Fab-1 were identified by determining the specific GEE-labeled residues that exhibited a reduction in the rate of labeling after complex formation. A significant reduction in the rate of GEE labeling was observed for E93 in the VEGF tryptic peptide V5, and D28 and E57 in the Fab-1 tryptic peptides HC2 and HC4, respectively. Results from the carboxyl group footprinting were compared with the binding interface identified from a previously characterized crystal structure (PDB: 1BJ1). All of these residues are located at the Fab-1:VEGF interface according to the crystal structure, demonstrating the potential utility of carboxyl group footprinting with GEE labeling for mapping epitopes.

  3. A KAS2 cDNA complements the phenotypes of the Arabidopsis fab1 mutant that differs in a single residue bordering the substrate binding pocket

    DEFF Research Database (Denmark)

    Carlsson, A.S.; LaBrie, S.T.; Kinney, A.J.;


    The fab1 mutant of Arabidopsis is partially deficient in activity of ß-ketoacyl-[acyl carrier protein] synthase II (KAS II). This defect results in increased levels of 16 : 0 fatty acid and is associated with damage and death of the mutants at low temperature. Transformation of fab1 plants with a c...... chain to bend. For functional analysis the equivalent Leu207Phe mutation was introduced into the fabB gene encoding the E. coli KAS I enzyme. Compared to wild-type, the Leu207Phe protein showed a 10-fold decrease in binding affinity for the fatty acid substrate, exhibited a modified behavior during size...


    African Journals Online (AJOL)

    STUDIES OF RHIZOBIUM INOCULATION AND FERTILIZER. TREATMENT ..... Number! in the same column followed by different letters are significantly different at 5% level (Duncan's multiple range ... However, in most cases both T5 and T6 ...

  5. 77 FR 31643 - Siltronic Corporation FAB1 Plant Including On-Site Leased Workers From Express Temporaries... (United States)


    ... on April 27, 2012 (77 FR 25201). The workers were engaged in the production of silicon wafers. At the... workers leased from the afore-mentioned agencies who work(ed) on-site at subject firm. The amended...

  6. Ste12/Fab1 phosphatidylinositol-3-phosphate 5-kinase is required for nitrogen-regulated mitotic commitment and cell size control (United States)

    Schauries, Marie; Kaczmarek, Adrian; Franz-Wachtel, Mirita; Du, Wei; Krug, Karsten; Maček, Boris; Petersen, Janni


    Tight coupling of cell growth and cell cycle progression enable cells to adjust their rate of division, and therefore size, to the demands of proliferation in varying nutritional environments. Nutrient stress promotes inhibition of Target Of Rapamycin Complex 1 (TORC1) activity. In fission yeast, reduced TORC1 activity advances mitotic onset and switches growth to a sustained proliferation at reduced cell size. A screen for mutants, that failed to advance mitosis upon nitrogen stress, identified a mutant in the PIKFYVE 1-phosphatidylinositol-3-phosphate 5-kinase fission yeast homolog Ste12. Ste12PIKFYVE deficient mutants were unable to advance the cell cycle to reduce cell size after a nitrogen downshift to poor nitrogen (proline) growth conditions. While it is well established that PI(3,5)P2 signalling is required for autophagy and that Ste12PIKFYVE mutants have enlarged vacuoles (yeast lysosomes), neither a block to autophagy or mutants that independently have enlarged vacuoles had any impact upon nitrogen control of mitotic commitment. The addition of rapamycin to Ste12PIKFYVE deficient mutants reduced cell size at division to suggest that Ste12PIKFYVE possibly functions upstream of TORC1. ste12 mutants display increased Torin1 (TOR inhibitor) sensitivity. However, no major impact on TORC1 or TORC2 activity was observed in the ste12 deficient mutants. In summary, Ste12PIKFYVE is required for nitrogen-stress mediated advancement of mitosis to reduce cell size at division. PMID:28273166

  7. Dicty_cDB: Contig-U06430-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available day-old yolk-sac larvae(trunk)... 44 5.2 1 ( DY515520 ) sh3P0031I22_F.ab1 adult sheep fracture... callus 10d... 44 5.2 1 ( DY508798 ) sh3P0010H18_F.ab1 adult sheep fracture callus 10d... 44 5....2 1 ( DY504087 ) sh2P0049F04_F.ab1 adult sheep fracture callus 7d ... 44 5.2 1 ( DY502969 ) sh2P0045H20_F.ab1 adult sheep fracture... callus 7d ... 44 5.2 1 ( DY500202 ) sh2P0036P17_F.ab1 adult sheep fracture... callus 7d ... 44 5.2 1 ( DY495721 ) sh2P0016N06_F.ab1 adult sheep fracture callus 7d ... 4

  8. Dicty_cDB: Contig-U01482-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 46A07_F.ab1 adult sheep fracture callus 10d... 50 0.050 1 ( DY519768 ) sh3P0045B11_F.ab1 adult sheep fracture... callus 10d... 50 0.050 1 ( DY511140 ) sh3P0017I22_F.ab1 adult sheep fracture ca...llus 10d... 50 0.050 1 ( DY499072 ) sh2P0033D24_F.ab1 adult sheep fracture callus 7d ... 50 0.050 1 ( DY4990...68 ) sh2P0033D18_F.ab1 adult sheep fracture callus 7d ... 50 0.050 1 ( DY497274 ) sh2P0021I13_F.ab1 adult sheep fracture... callus 7d ... 50 0.050 1 ( DY490007 ) sh1-we03_l0.s2 adult sheep fracture callus pool O... 50 0

  9. Reference: 563 [Arabidopsis Phenome Database[Archive

    Lifescience Database Archive (English)

    Full Text Available demonstrating that fab1-1 encodes an active KASII. Strong seed-specific hairpin-RNAi reductions in FAB1 expression resulted in abort...ion of approximately 1/4 of the embryos in an apparent p

  10. Dicty_cDB: Contig-U15643-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 11152.CR_N11 GC_BGC-11 Bos taurus cDNA clone I... 40 0.024 2 ( DY517183 ) sh3P0036P15_F.ab1 adult sheep fracture...4 0.034 2 ( DY517185 ) sh3P0036P17_F.ab1 adult sheep fracture callus 10d... 40 0.034 2 ( DY135204 ) 010128BE

  11. Physical functional outcome assessment of patients with major burns admitted to a UK Burn Intensive Care Unit. (United States)

    Smailes, Sarah T; Engelsman, Kayleen; Dziewulski, Peter


    Determining the discharge outcome of burn patients can be challenging and therefore a validated objective measure of functional independence would assist with this process. We developed the Functional Assessment for Burns (FAB) score to measure burn patients' functional independence. FAB scores were taken on discharge from ICU (FAB 1) and on discharge from inpatient burn care (FAB 2) in 56 patients meeting the American Burn Association criteria for major burn. We retrospectively analysed prospectively collected data to measure the progress of patients' physical functional outcomes and to evaluate the predictive validity of the FAB score for discharge outcome. Mean age was 38.6 years and median burn size 35%. Significant improvements were made in the physical functional outcomes between FAB 1 and FAB 2 scores (ppatients were discharged home, 8 of these with social care. 8 patients were transferred to another hospital for further inpatient rehabilitation. FAB 1 score (≤ 9) is strongly associated with discharge outcome (pburn patients.

  12. Native MS and ECD Characterization of a Fab-Antigen Complex May Facilitate Crystallization for X-ray Diffraction (United States)

    Zhang, Ying; Cui, Weidong; Wecksler, Aaron T.; Zhang, Hao; Molina, Patricia; Deperalta, Galahad; Gross, Michael L.


    Native mass spectrometry (MS) and top-down electron-capture dissociation (ECD) combine as a powerful approach for characterizing large proteins and protein assemblies. Here, we report their use to study an antibody Fab (Fab-1)-VEGF complex in its near-native state. Native ESI with analysis by FTICR mass spectrometry confirms that VEGF is a dimer in solution and that its complex with Fab-1 has a binding stoichiometry of 2:2. Applying combinations of collisionally activated dissociation (CAD), ECD, and infrared multiphoton dissociation (IRMPD) allows identification of flexible regions of the complex, potentially serving as a guide for crystallization and X-ray diffraction analysis.

  13. Dicty_cDB: Contig-U16424-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 19K01_F.ab1 adult sheep fracture callus 7d ... 40 0.72 3 ( CU654057 ) ovine foetal ovary tissues (Prophase I...sbb001f193d17f0 Sorghum methylation filtered lib... 34 0.70 2 ( DY496661 ) sh2P00

  14. Dicty_cDB: Contig-U03667-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available re callus 7d ... 46 1.6 1 ( DY490191 ) sh1-xb10_l0.s2 adult sheep fracture callus p...9OML22049076HT OML2 Ovis aries cDNA, mRNA se... 46 1.6 1 ( DY496264 ) sh2P0018H02_F.ab1 adult sheep fractu

  15. Human combinatorial Fab library yielding specific and functional antibodies against the human fibroblast growth factor receptor 3. (United States)

    Rauchenberger, Robert; Borges, Eric; Thomassen-Wolf, Elisabeth; Rom, Eran; Adar, Rivka; Yaniv, Yael; Malka, Michael; Chumakov, Irina; Kotzer, Sarit; Resnitzky, Dalia; Knappik, Achim; Reiffert, Silke; Prassler, Josef; Jury, Karin; Waldherr, Dirk; Bauer, Susanne; Kretzschmar, Titus; Yayon, Avner; Rothe, Christine


    The human combinatorial antibody library Fab 1 (HuCAL-Fab 1) was generated by transferring the heavy and light chain variable regions from the previously constructed single-chain Fv library (Knappik, A., Ge, L., Honegger, A., Pack, P., Fischer, M., Wellnhofer, G., Hoess, A., Wölle, J., Plückthun, A., and Virnekäs, B. (2000) J. Mol. Biol. 296, 57-86), diversified in both complementarity-determining regions 3 into a novel Fab display vector, yielding 2.1 x 10(10) different antibody fragments. The modularity has been retained in the Fab display and screening plasmids, ensuring rapid conversion into various antibody formats as well as antibody optimization using prebuilt maturation cassettes. HuCAL-Fab 1 was challenged against the human fibroblast growth factor receptor 3, a potential therapeutic antibody target, against which, to the best of our knowledge, no functional antibodies could be generated so far. A unique screening mode was designed utilizing recombinant functional proteins and cell lines differentially expressing fibroblast growth factor receptor isoforms diversified in expression and receptor dependence. Specific Fab fragments with subnanomolar affinities were isolated by selection without any maturation steps as determined by fluorescence flow cytometry. Some of the selected Fab fragments completely inhibit target-mediated cell proliferation, rendering them the first monoclonal antibodies against fibroblast growth factor receptors having significant function blocking activity. This study validates HuCAL-Fab 1 as a valuable source for the generation of target-specific antibodies for therapeutic applications.

  16. Dicty_cDB: Contig-U10844-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 3P06_F.ab1 adult sheep fracture callus 10d... 48 0.55 1 ( DT327029 ) SusFleck_BF_0013_0016_5_pCNS_N12 SusFle.... 48 0.55 1 ( AK046564 ) Mus musculus 4 days neonate male adipose cDNA, RI... 48 0.55 1 ( DY509951 ) sh3P001

  17. Increasing the utility of the Functional Assessment for Burns Score: Not just for major burns. (United States)

    Smailes, Sarah T; Engelsman, Kayleen; Rodgers, Louise; Upson, Clara


    The Functional Assessment for Burns (FAB) score is established as an objective measure of physical function that predicts discharge outcome in adult patients with major burn. However, its validity in patients with minor and moderate burn is unknown. This is a multi-centre evaluation of the predictive validity of the FAB score for discharge outcome in adult inpatients with minor and moderate burns. FAB assessments were undertaken within 48 h of admission to (FAB 1), and within 48 h of discharge (FAB 2) from burn wards in 115 patients. Median age was 45 years and median burn size 4%. There were significant improvements in the patients' FAB scores (pburns.

  18. Deleción terminal del brazo largo del cromosoma 9 en Leucemia promielocítica aguda



    La Leucemia Promielocítica Aguda (LPA), definida como M3 en la clasificación francesa-americana- británica (FAB)(1), se caracteriza por presentar una translocación cromosómica al azar t(15;17)(q22;q21). En esta translocación se fusionan los receptores del ácido retinoíco ?(? RAR) gen localizado en el cromosoma 17 y el gen PML sobre el cromosoma 15(2). Por otra lado, la deleción del brazo largo del cromosoma 9(9q-) es una alteración rara específica encontrada en (LMA) como una anormalidad únic...

  19. Genomic approaches towards identification of components involved in peptide based cell growth of Arabidopsis thailana

    DEFF Research Database (Denmark)

    Mahmood, Khalid

    Secreted peptides are considered now as important signaling molecules involved in plant growth and development. Plant peptide containing sulfated tyrosine 1 (PSY1) is a small peptide that promotes cell elongation and expansion at nanomolar concentration. This is achieved through binding...... elongation. FAB1C is highly down regulated in psy1r mutant plants and is assumed to play role in acidification and formation of vacuole that may contribute in cell elongation. In short, our work provides insights how growth coordinated through cellular communication using PSY1 as a signal molecule....... to extracellular domain of the leucine-rich repeat (LRR) receptor kinase called PSY1R. Upon binding of the peptide, PSY1R transduces the signal by phosphorylating the plasma membrane H+-ATPase (AHA2) leading to proton extrusion results in cell elongation. To understand the molecular basis of PSY1 response...

  20. Differential Gene Expression and Protein Phosphorylation as Factors Regulating the State of the Arabidopsis SNX1 Protein Complexes in Response to Environmental Stimuli (United States)

    Brumbarova, Tzvetina; Ivanov, Rumen


    Endosomal recycling of plasma membrane proteins contributes significantly to the regulation of cellular transport and signaling processes. Members of the Arabidopsis (Arabidopsis thaliana) SORTING NEXIN (SNX) protein family were shown to mediate the endosomal retrieval of transporter proteins in response to external challenges. Our aim is to understand the possible ways through which external stimuli influence the activity of SNX1 in the root. Several proteins are known to contribute to the function of SNX1 through direct protein–protein interaction. We, therefore, compiled a list of all Arabidopsis proteins known to physically interact with SNX1 and employed available gene expression and proteomic data for a comprehensive analysis of the transcriptional and post-transcriptional regulation of this interactome. The genes encoding SNX1-interaction partners showed distinct expression patterns with some, like FAB1A, being uniformly expressed, while others, like MC9 and BLOS1, were expressed in specific root zones and cell types. Under stress conditions known to induce SNX1-dependent responses, two genes encoding SNX1-interacting proteins, MC9 and NHX6, showed major gene-expression variations. We could also observe zone-specific transcriptional changes of SNX1 under iron deficiency, which are consistent with the described role of the SNX1 protein. This suggests that the composition of potential SNX1-containing protein complexes in roots is cell-specific and may be readjusted in response to external stimuli. On the level of post-transcriptional modifications, we observed stress-dependent changes in the phosphorylation status of SNX1, FAB1A, and CLASP. Interestingly, the phosphorylation events affecting SNX1 interactors occur in a pattern which is largely complementary to transcriptional regulation. Our analysis shows that transcriptional and post-transcriptional regulation play distinct roles in SNX1-mediated endosomal recycling under external stress. PMID:27725825

  1. Development of capillary size exclusion chromatography for the analysis of monoclonal antibody fragments extracted from human vitreous humor. (United States)

    Rea, Jennifer C; Lou, Yun; Cuzzi, Joel; Hu, Yuhua; de Jong, Isabella; Wang, Yajun Jennifer; Farnan, Dell


    Recombinant antigen-binding fragments (Fabs) are currently on the market and in development for the treatment of ophthalmologic indications. Recently, Quality by Design (QbD) initiatives have been implemented that emphasize understanding the relationship between quality attributes of the product and their impact on safety and efficacy. In particular, changes in product quality once the protein is administered to the patient are of particular interest. Knowledge of protein aggregation in vivo is of importance due to the possibility of antibody aggregates eliciting an immunogenic response in the patient. Presently, there are few analytical methods with adequate sensitivity to analyze Fab aggregates in human vitreous humor (HVH) because the Fab amount available for analysis is often quite low. Here, we report the development of a highly sensitive capillary size exclusion chromatography (SEC) methodology for Fab aggregate analysis in HVH. We demonstrate a process to perform capillary SEC to analyze Fabs with picogram sensitivity and an RSD of less than 8% for the relative peak area of high molecular weight species (HMWS). In addition, we have developed a Protein G affinity chromatography method to capture Fabs from HVH for capillary SEC analysis. Recovery efficiencies ranging from 86 to 99% were achieved using this recovery method with 300 μL HVH samples containing Fab1. Finally, we demonstrate the applicability of the methodology by quantifying Fab aggregates in HVH, which can potentially be used for aggregate analysis of clinically relevant samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Deleción terminal del brazo largo del cromosoma 9 en Leucemia promielocítica aguda

    Directory of Open Access Journals (Sweden)

    Herrera JC.


    Full Text Available La Leucemia Promielocítica Aguda (LPA, definida como M3 en la clasificación francesa-americana- británica (FAB(1, se caracteriza por presentar una translocación cromosómica al azar t(15;17(q22;q21. En esta translocación se fusionan los receptores del ácido retinoíco ?(? RAR gen localizado en el cromosoma 17 y el gen PML sobre el cromosoma 15(2. Por otra lado, la deleción del brazo largo del cromosoma 9(9q- es una alteración rara específica encontrada en (LMA como una anormalidad única del cariotipo o como un cambio secundario, particularmente junto con t(8;21(q22;q22(3,4. La mayoría de ellas son deleciones intersticiales, 9q12 y 9q22; constituyen el sitio más común de ruptura proximal y distal (5.

  3. Lysosomal interaction of Akt with Phafin2: a critical step in the induction of autophagy.

    Directory of Open Access Journals (Sweden)

    Mami Matsuda-Lennikov

    Full Text Available Autophagy is an evolutionarily conserved mechanism for the gross disposal of intracellular proteins in mammalian cells and dysfunction in this pathway has been associated with human disease. Although the serine threonine kinase Akt is suggested to play a role in this process, little is known about the molecular mechanisms by which Akt induces autophagy. Using a yeast two-hybrid screen, Phafin2 (EAPF or PLEKHF2, a lysosomal protein with a unique structure of N-terminal PH (pleckstrin homology domain and C-terminal FYVE (Fab 1, YOTB, Vac 1, and EEA1 domain was found to interact with Akt. A sucrose gradient fractionation experiment revealed that both Akt and Phafin2 co-existed in the same lysosome enriched fraction after autophagy induction. Confocal microscopic analysis and BiFC analysis demonstrated that both Akt and Phafin2 accumulate in the lysosome after induction of autophagy. BiFC analysis using PtdIns (3P interaction defective mutant of Phafin2 demonstrated that lysosomal accumulation of the Akt-Phafin2 complex and subsequent induction of autophagy were lysosomal PtdIns (3P dependent events. Furthermore, in murine macrophages, both Akt and Phafin2 were required for digestion of fluorescent bacteria and/or LPS-induced autophagy. Taken together, these findings establish that lysosomal accumulation of Akt and Phafin2 is a critical step in the induction of autophagy via an interaction with PtdIns (3P.

  4. Mask cycle time reduction for foundry projects (United States)

    Balasinski, A.


    One of key deliverables of foundry based manufacturing is low cycletime. Building new and enhancing existing products by mask changes involves significant logistical effort, which could be reduced by standardizing data management and communication procedures among design house, mask shop, and foundry (fab) [1]. As an example, a typical process of taping out can take up to two weeks in addition to technical effort, for database handling, mask form completion, management approval, PO signoff and JDV review, translating into loss of revenue. In order to reduce this delay, we are proposing to develop a unified online system which should assist with the following functions: database edits, final verifications, document approvals, mask order entries, and JDV review with engineering signoff as required. This would help a growing number of semiconductor products to be flexibly manufactured at different manufacturing sites. We discuss how the data architecture based on a non-relational database management system (NRDMBS) extracted into a relational one (RDMBS) should provide quality information [2], to reduce cycle time significantly beyond 70% for an example 2 week tapeout schedule.

  5. Elastic-contractile model proteins: Physical chemistry, protein function and drug design and delivery. (United States)

    Urry, Dan W; Urry, Kelley D; Szaflarski, Witold; Nowicki, Michal


    This review presents the structure and physico-chemical properties of ECMPs, elastic-contractile model proteins using sparse design modifications of elastic (GVGVP)(n); it describes the capacity of ECMP to perform the energy conversions that sustain living organisms; it arrives at the hydration thermodynamics of ECMP in terms of the change in Gibbs free energy of hydrophobic association, ΔG(HA), and the apolar-polar repulsive free energy of hydration, ΔG(ap); it applies ΔG(HA), ΔG(ap), and the nature of elasticity to describe the function of basic diverse proteins, namely - the F₁-motor of ATP synthase, Complex III of mitochondria, the KscA potassium-channel, and the molecular chaperonin, GroEL/ES; it applies ΔG(HA) and ΔG(ap) to describe the function of ABC exporter proteins that confer multi-drug resistance (MDR) on micro-organisms and human carcinomas and suggests drug modifications with which to overcome MDR. Using ECMP, means are demonstrated, for quantifying drug hydrophobicity with which to combat MDR and for preparing ECMP drug delivery nanoparticles, ECMPddnp, decorated with synthetic antigen-binding fragments, Fab1 and Fab2, with which to target specific up-regulated receptors, characteristic of human carcinoma cells, for binding and localized drug release. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. Identification of Genes Affecting Vacuole Membrane Fragmentation in Saccharomyces cerevisiae (United States)

    Michaillat, Lydie; Mayer, Andreas


    The equilibrium of membrane fusion and fission influences the volume and copy number of organelles. Fusion of yeast vacuoles has been well characterized but their fission and the mechanisms determining vacuole size and abundance remain poorly understood. We therefore attempted to systematically characterize factors necessary for vacuole fission. Here, we present results of an in vivo screening for deficiencies in vacuolar fragmentation activity of an ordered collection deletion mutants, representing 4881 non-essential genes of the yeast Saccharomyces cerevisiae. The screen identified 133 mutants with strong defects in vacuole fragmentation. These comprise numerous known fragmentation factors, such as the Fab1p complex, Tor1p, Sit4p and the V-ATPase, thus validating the approach. The screen identified many novel factors promoting vacuole fragmentation. Among those are 22 open reading frames of unknown function and three conspicuous clusters of proteins with known function. The clusters concern the ESCRT machinery, adaptins, and lipases, which influence the production of diacylglycerol and phosphatidic acid. A common feature of these factors of known function is their capacity to change membrane curvature, suggesting that they might promote vacuole fragmentation via this property. PMID:23383298

  7. Proteomics for Drug Resistance on the Food Chain? Multidrug-Resistant Escherichia coli Proteomes from Slaughtered Pigs. (United States)

    Ramos, Sónia; Silva, Nuno; Hébraud, Michel; Santos, Hugo M; Nunes-Miranda, Júlio Dinis; Pinto, Luís; Pereira, José E; Capelo, José-Luis; Poeta, Patrícia; Igrejas, Gilberto


    Understanding global drug resistance demands an integrated vision, focusing on both human and veterinary medicine. Omics technologies offer new vistas to decipher mechanisms of drug resistance in the food chain. For example, Escherichia coli resistance to major antibiotics is increasing whereas multidrug resistance (MDR) strains are now commonly found in humans and animals. Little is known about the structural and metabolic changes in the cell that trigger resistance to antimicrobial agents. Proteomics is an emerging field that is used to advance our knowledge in global health and drug resistance in the food chain. In the present proteomic analysis, we offer an overview of the global protein expression of different MDR E. coli strains from fecal samples of pigs slaughtered for human consumption. A full proteomic survey of the drug-resistant strains SU60, SU62, SU76, and SU23, under normal growth conditions, was made by two-dimensional electrophoresis, identifying proteins by MALDI-TOF/MS. The proteomes of these four E. coli strains with different genetic profiles were compared in detail. Identical transport, stress response, or metabolic proteins were discovered in the four strains. Several of the identified proteins are essential in bacterial pathogenesis (GAPDH, LuxS, FKBPs), development of bacterial resistance (Omp's, TolC, GroEL, ClpB, or SOD), and potential antibacterial targets (FBPA, FabB, ACC's, or Fab1). Effective therapies against resistant bacteria are crucial and, to accomplish this, a comprehensive understanding of putative resistance mechanisms is essential. Moving forward, we suggest that multi-omics research will further improve our knowledge about bacterial growth and virulence on the food chain, especially under antibiotic stress.

  8. Unusual Water-mediated Antigenic Recognition of the Proinflammatory Cytokine Interleukin-18

    Energy Technology Data Exchange (ETDEWEB)

    Argiriadi, Maria A.; Xiang, Tao; Wu, Chengbin; Ghayur, Tariq; Borhani, David W.; (Abbott)


    The unique cytokine interleukin-18 (IL-18) acts synergistically with IL-12 to regulate T-helper 1 and 2 lymphocytes and, as such, seems to underlie the pathogenesis of various autoimmune and allergic diseases. Several anti-IL-18 agents are in clinical development, including the recombinant human antibody ABT-325, which is entering trials for autoimmune diseases. Given competing cytokine/receptor and cytokine/receptor decoy interactions, understanding the structural basis for recognition is critical for effective development of anti-cytokine therapies. Here we report three crystal structures: the murine antibody 125-2H Fab fragment bound to human IL-18, at 1.5 {angstrom} resolution; the 125-2H Fab (2.3 {angstrom}); and the ABT-325 Fab (1.5 {angstrom}). These structures, along with human/mouse IL-18 chimera binding data, allow us to make three key observations relevant to the biology and antigenic recognition of IL-18 and related cytokines. First, several IL-18 residues shift dramatically (>10 {angstrom}) upon binding 125-2H, compared with unbound IL-18 (Kato, Z., Jee, J., Shikano, H., Mishima, M., Ohki, I., Ohnishi, H., Li, A., Hashimoto, K., Matsukuma, E., Omoya, K., Yamamoto, Y., Yoneda, T., Hara, T., Kondo, N., and Shirakawa, M. (2003) Nat. Struct. Biol. 10, 966-971). IL-18 thus exhibits plasticity that may be common to its interactions with other receptors. Related cytokines may exhibit similar plasticity. Second, ABT-325 and 125-2H differ significantly in combining site character and architecture, thus explaining their ability to bind IL-18 simultaneously at distinct epitopes. These data allow us to define the likely ABT-325 epitope and thereby explain the distinct neutralizing mechanisms of both antibodies. Third, given the high 125-2H potency, 10 well ordered water molecules are trapped upon complex formation in a cavity between two IL-18 loops and all six 125-2H complementarity-determining regions. Thus, counterintuitively, tight and specific antibody

  9. Enabling inspection solutions for future mask technologies through the development of massively parallel E-Beam inspection (United States)

    Malloy, Matt; Thiel, Brad; Bunday, Benjamin D.; Wurm, Stefan; Jindal, Vibhu; Mukhtar, Maseeh; Quoi, Kathy; Kemen, Thomas; Zeidler, Dirk; Eberle, Anna Lena; Garbowski, Tomasz; Dellemann, Gregor; Peters, Jan Hendrik


    The new device architectures and materials being introduced for sub-10nm manufacturing, combined with the complexity of multiple patterning and the need for improved hotspot detection strategies, have pushed current wafer inspection technologies to their limits. In parallel, gaps in mask inspection capability are growing as new generations of mask technologies are developed to support these sub-10nm wafer manufacturing requirements. In particular, the challenges associated with nanoimprint and extreme ultraviolet (EUV) mask inspection require new strategies that enable fast inspection at high sensitivity. The tradeoffs between sensitivity and throughput for optical and e-beam inspection are well understood. Optical inspection offers the highest throughput and is the current workhorse of the industry for both wafer and mask inspection. E-beam inspection offers the highest sensitivity but has historically lacked the throughput required for widespread adoption in the manufacturing environment. It is unlikely that continued incremental improvements to either technology will meet tomorrow's requirements, and therefore a new inspection technology approach is required; one that combines the high-throughput performance of optical with the high-sensitivity capabilities of e-beam inspection. To support the industry in meeting these challenges SUNY Poly SEMATECH has evaluated disruptive technologies that can meet the requirements for high volume manufacturing (HVM), for both the wafer fab [1] and the mask shop. Highspeed massively parallel e-beam defect inspection has been identified as the leading candidate for addressing the key gaps limiting today's patterned defect inspection techniques. As of late 2014 SUNY Poly SEMATECH completed a review, system analysis, and proof of concept evaluation of multiple e-beam technologies for defect inspection. A champion approach has been identified based on a multibeam technology from Carl Zeiss. This paper includes a discussion on the

  10. 大容量人免疫球蛋白G基因文库的构建及初步应用%Construction and preliminary use of a large human IgG gene library

    Institute of Scientific and Technical Information of China (English)

    邵红霞; 付永锋; Hiroshi Tachibana; 程训佳


    Objective To produce human monoclonal antibody Fab fragments specific to group 2 mite allergens from a combinatorial immunoglobulin gene library. Methods Total genes encoding the light chains and Fd region of the heavy chains of human immunoglobulin G were amplified from the peripheral blood lymphocytes of 300 healthy volunteers by re-combinant DNA technology. A human IgG gene library was constructed by inserting the light and heavy chain genes into the vector pFabl-His2. The library was screened for the production of human monoclonal antibody Fab fragments with an affinity for Der f2 by cloning blotting assay and results were confirmed by ELISA. Genes of positive clones were identified and the amino acid sequences were deduced from the DNA sequences; the variable regions of the heavy and light chain were compared with a Kabat database. Affinity measurements of purified Fabs by surface plasmon resonance were carried out using the BIAcore 3000. Results After about 1. 28×10~6 clones were screened, 2 positive Fab fragments specific to rDer f2 named AMI and AM2 were identified. The heavy chains of both clones were completely homologous. Sequence a-nalysis of both AMI and AM2 light chain genes revealed that the nearest V-segment germlines were K2-30 and K1-12, respectively. The closest V-segment germline of the heavy chain gene was VH3 -30. Plasmon resonance assay revealed that the association and dissociation constants were 6. 1×10~7M~(-1) and 3. 1×10~(-8)M for AMI and 5. 7×10~7M~(-1) and 4. 1×10~8 M for AM2, respectively. Conclusion High-affinity human monoclonal antibody Fab fragments specific to mite allergens were obtained from a large naive combinatorial library of immunoglobulin genes.%目的 从大容量人免疫球蛋白G基因文库中获得尘螨2类变应原特异的IgG Fab片段. 方法 通过DNA重组技术,从300名健康志愿者的外周血淋巴细胞中扩增全套人抗体轻链及重链Fd基因,分别插入pFab1-His2相应位置,构建人免