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Sample records for prolyl oligopeptidase substrates

  1. Prolyl Oligopeptidase from the Blood Fluke Schistosoma mansoni: From Functional Analysis to Anti-schistosomal Inhibitors

    Czech Academy of Sciences Publication Activity Database

    Fajtová, P.; Štefanić, S.; Hradilek, M.; Dvořák, Jan; Vondrášek, J.; Jílková, A.; Ulrychová, L.; McKerrow, J.H.; Caffrey, C.R.; Mareš, M.; Horn, M.

    2015-01-01

    Roč. 9, č. 6 (2015), e0003827 ISSN 1935-2735 Institutional support: RVO:60077344 Keywords : Schistosoma mansoni * schistosomiasis * prolyl oligopeptidase * blood fluke Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.948, year: 2015

  2. Molecular dynamics study of prolyl oligopeptidase with inhibitor in binding cavity

    NARCIS (Netherlands)

    Kaszuba, K.; Róg, T.; St-Pierre, J.-F.; Männistö, P.T.; Karttunen, M.E.J.; Bunker, A.

    2009-01-01

    We used the crystal structure of prolyl oligopeptidase (POP) with bound Z-pro-prolinal (ZPP) inhibitor (Protein Data Bank (PDB) structure 1QFS) to perform an intensive molecular dynamics study of the POP-ZPP complex. We performed 100 ns of simulation with the hemiacetal bond, through which the ZPP

  3. Prolyl Oligopeptidase from the Blood Fluke Schistosoma mansoni: From Functional Analysis to Anti-schistosomal Inhibitors

    Czech Academy of Sciences Publication Activity Database

    Fajtová, Pavla; Štefanic, S.; Hradilek, Martin; Dvořák, Jan; Vondrášek, Jiří; Jílková, Adéla; Ulrychová, Lenka; McKerrow, J. H.; Caffrey, C. R.; Mareš, Michael; Horn, Martin

    2015-01-01

    Roč. 9, č. 6 (2015), e0003827/1-e0003827/24 ISSN 1935-2735 R&D Projects: GA ČR(CZ) GAP302/11/1481; GA MŠk LO1302 Institutional support: RVO:61388963 ; RVO:68378050 Keywords : Schistosoma mansoni * schistosomiasis * prolyl oligopeptidase * blood fluke Subject RIV: CE - Biochemistry; EB - Genetics ; Molecular Biology (UMG-J) Impact factor: 3.948, year: 2015 http://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0003827

  4. Prolyl oligopeptidase and dipeptidyl peptidase II/dipeptidyl peptidase IV ratio in the cerebrospinal fluid in Parkinson's disease: historical overview and future prospects.

    Science.gov (United States)

    Nagatsu, Toshiharu

    2017-06-01

    Prolyl oligopeptidase (also named prolyl endopeptidase; PREP) hydrolyzes the Pro-Xaa bonds of biologically active oligopeptides on their carboxyl side. In 1987, we detected PREP activity in human cerebrospinal fluid (CSF) using highly sensitive liquid chromatography-fluorometry with succinyl-Gly-Pro-4-methyl-coumarin amide as a new synthetic substrate, and found a marked decrease in its activity in the cerebrospinal fluid (CSF) from patients with Parkinson's disease (PD) as compared with its level in control patients without neurological diseases. In 2013, Hannula et al. found co-localization of PREP with α-synuclein in the postmortem PD brain. Several recent studies also suggest that the level of PREP in the brain of PD patients may be related to dopamine (DA) cell death via promotion of α-synuclein oligomerization and that inhibitors of PREP may play a neuroprotective role in PD. Although the relationship between another family of prolyl oligopeptidase enzymes, dipeptidyl peptidase II (DPP II) and dipeptidyl peptidase IV (DPP IV), and α-synuclein in the PD brain is not yet clear, we found that the DPP II activity/DPP IV activity ratio in the CSF was significantly increased in PD patients. This review discusses the possibility of PREP as well as the DPP II/DPP IV ratio in the CSF as potential biomarkers of PD.

  5. Prolyl oligopeptidase inhibition-induced growth arrest of human gastric cancer cells

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    Suzuki, Kanayo [Laboratory of Cell Biology, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094 (Japan); Sakaguchi, Minoru, E-mail: sakaguti@gly.oups.ac.jp [Laboratory of Cell Biology, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094 (Japan); Tanaka, Satoshi [Laboratory of Cell Biology, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094 (Japan); Yoshimoto, Tadashi [Department of Life Science, Setsunan University, 17-8 Ikeda-Nakamachi, Neyagawa, Osaka 572-8508 (Japan); Takaoka, Masanori [Laboratory of Cell Biology, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094 (Japan)

    2014-01-03

    Highlights: •We examined the effects of prolyl oligopeptidase (POP) inhibition on p53 null gastric cancer cell growth. •POP inhibition-induced cell growth suppression was associated with an increase in a quiescent G{sub 0} state. •POP might regulate the exit from and/or reentry into the cell cycle. -- Abstract: Prolyl oligopeptidase (POP) is a serine endopeptidase that hydrolyzes post-proline peptide bonds in peptides that are <30 amino acids in length. We recently reported that POP inhibition suppressed the growth of human neuroblastoma cells. The growth suppression was associated with pronounced G{sub 0}/G{sub 1} cell cycle arrest and increased levels of the CDK inhibitor p27{sup kip1} and the tumor suppressor p53. In this study, we investigated the mechanism of POP inhibition-induced cell growth arrest using a human gastric cancer cell line, KATO III cells, which had a p53 gene deletion. POP specific inhibitors, 3-((4-[2-(E)-styrylphenoxy]butanoyl)-L-4-hydroxyprolyl)-thiazolidine (SUAM-14746) and benzyloxycarbonyl-thioprolyl-thioprolinal, or RNAi-mediated POP knockdown inhibited the growth of KATO III cells irrespective of their p53 status. SUAM-14746-induced growth inhibition was associated with G{sub 0}/G{sub 1} cell cycle phase arrest and increased levels of p27{sup kip1} in the nuclei and the pRb2/p130 protein expression. Moreover, SUAM-14746-mediated cell cycle arrest of KATO III cells was associated with an increase in the quiescent G{sub 0} state, defined by low level staining for the proliferation marker, Ki-67. These results indicate that POP may be a positive regulator of cell cycle progression by regulating the exit from and/or reentry into the cell cycle by KATO III cells.

  6. Inhibitory effect of verbascoside isolated from Buddleja brasiliensis Jacq. ex Spreng on prolyl oligopeptidase activity.

    Science.gov (United States)

    Filho, Augusto G; Morel, Ademir F; Adolpho, Luciana; Ilha, Vinícius; Giralt, Ernest; Tarragó, Teresa; Dalcol, Ionara I

    2012-10-01

    The phenylpropanoid glycoside verbascoside [2-(3,4-dihydroxyphenylethyl)-1-O-α-L-rhamnopyranosyl-(1→3)-β-D-(4-O-caffeyl)-glucopyranoside] (1) has been isolated as the main constituent of the crude extract of Buddleja brasiliensis Jacq. ex Spreng from Southern Brazil. The crude extract, main fractions and the compound 1 were evaluated for inhibition of the enzymes acetylcholinesterase (AChE), dipeptidyl peptidase-IV (DPP-IV) and prolyl oligopeptidase (POP). Compound 1 showed weak activity against DPP-IV with an IC(50) > 150 µM and was inactive against AChE, with a pMIQ determined by bioautography of 9.6. In contrast, 1 displayed significant inhibition of POP in a dose-dependent manner with an IC(50) value of 1.3 ± 0.2 µM, similar to the positive control, baicalin, with a POP IC(50) of 12 ± 3 µM. Copyright © 2012 John Wiley & Sons, Ltd.

  7. Structural analysis of prolyl oligopeptidases using molecular docking and dynamics: insights into conformational changes and ligand binding.

    Directory of Open Access Journals (Sweden)

    Swati Kaushik

    Full Text Available Prolyl oligopeptidase (POP is considered as an important pharmaceutical target for the treatment of numerous diseases. Despite enormous studies on various aspects of POPs structure and function still some of the questions are intriguing like conformational dynamics of the protein and interplay between ligand entry/egress. Here, we have used molecular modeling and docking based approaches to unravel questions like differences in ligand binding affinities in three POP species (porcine, human and A. thaliana. Despite high sequence and structural similarity, they possess different affinities for the ligands. Interestingly, human POP was found to be more specific, selective and incapable of binding to a few planar ligands which showed extrapolation of porcine POP in human context is more complicated. Possible routes for substrate entry and product egress were also investigated by detailed analyses of molecular dynamics (MD simulations for the three proteins. Trajectory analysis of bound and unbound forms of three species showed differences in conformational dynamics, especially variations in β-propeller pore size, which was found to be hidden by five lysine residues present on blades one and seven. During simulation, β-propeller pore size was increased by ∼2 Å in porcine ligand-bound form which might act as a passage for smaller product movement as free energy barrier was reduced, while there were no significant changes in human and A. thaliana POPs. We also suggest that these differences in pore size could lead to fundamental differences in mode of product egress among three species. This analysis also showed some functionally important residues which can be used further for in vitro mutagenesis and inhibitor design. This study can help us in better understanding of the etiology of POPs in several neurodegenerative diseases.

  8. Effects of a Natural Prolyl Oligopeptidase Inhibitor, Rosmarinic Acid, on Lipopolysaccharide-Induced Acute Lung Injury in Mice

    Directory of Open Access Journals (Sweden)

    Miaomiao Wei

    2012-03-01

    Full Text Available Rosmarinic acid (RA, a polyphenolic phytochemical, is a natural prolyl oligopeptidase inhibitor. In the present study, we found that RA exerted potent anti-inflammatory effects in in vivo models of acute lung injury (ALI induced by lipopolysaccharide (LPS. Mice were pretreated with RA one hour before challenge with a dose of 0.5 mg/kg LPS. Twenty-four hours after LPS was given, bronchoalveolar lavage fluid (BALF was obtained to measure pro-inflammatory mediator and total cell counts. RA significantly decreased the production of LPS-induced TNF-a, IL-6, and IL-1β compare with the LPS group. When pretreated with RA (5, 10, or 20 mg/kg the lung wet-to-dry weight (W/D ratio of the lung tissue and the number of total cells, neutrophils and macrophages in the BALF were decreased significantly. Furthermore, RA may enhance oxidase dimutase (SOD activity during the inflammatory response to LPS-induced ALI. And we further demonstrated that RA exerts anti-inflammation effect in vivo models of ALI through suppresses ERK/MAPK signaling in a dose dependent manner. These studies have important implications for RA administration as a potential treatment for ALI.

  9. Trypanosoma cruzi 80 kDa prolyl oligopeptidase (Tc80 as a novel immunogen for Chagas disease vaccine.

    Directory of Open Access Journals (Sweden)

    Augusto E Bivona

    2018-03-01

    Full Text Available Chagas disease, also known as American Trypanosomiasis, is a chronic parasitic disease caused by the flagellated protozoan Trypanosoma cruzi that affects about 8 million people around the world where more than 25 million are at risk of contracting the infection. Despite of being endemic on 21 Latin-American countries, Chagas disease has become a global concern due to migratory movements. Unfortunately, available drugs for the treatment have several limitations and they are generally administered during the chronic phase of the infection, when its efficacy is considered controversial. Thus, prophylactic and/or therapeutic vaccines are emerging as interesting control alternatives. In this work, we proposed Trypanosoma cruzi 80 kDa prolyl oligopeptidase (Tc80 as a new antigen for vaccine development against Chagas disease.In a murine model, we analyzed the immune response triggered by different immunization protocols based on Tc80 and evaluated their ability to confer protection against a challenge with the parasite. Immunized mice developed Tc80-specific antibodies which were able to carry out different functions such as: enzymatic inhibition, neutralization of parasite infection and complement-mediated lysis of trypomastigotes. Furthermore, vaccinated mice elicited strong cell-mediated immunity. Spleen cells from immunized mice proliferated and secreted Th1 cytokines (IL-2, IFN-γ and TNF-α upon re-stimulation with rTc80. Moreover, we found Tc80-specific polyfunctional CD4 T cells, and cytotoxic T lymphocyte activity against one Tc80 MHC-I peptide. Immunization protocols conferred protection against a T. cruzi lethal challenge. Immunized groups showed a decreased parasitemia and higher survival rate compared with non-immunized control mice. Moreover, during the chronic phase of the infection, immunized mice presented: lower levels of myopathy-linked enzymes, parasite burden, electrocardiographic disorders and inflammatory cells.Considering that

  10. A library of fluorescent peptides for exploring the substrate specificities of prolyl isomerases

    NARCIS (Netherlands)

    Zoldak, G.; Aumuller, T.; Lucke, C.; Hritz, J.; Oostenbrink, C.; Fischer, G.; Schmid, F.X.

    2009-01-01

    To fully explore the substrate specificities of prolyl isomerases, we synthesized a library of 20 tetrapeptides that are labeled with a 2-aminobenzoyl (Abz) group at the amino terminus and a p-nitroanilide (pNA) group at the carboxy terminus. In this peptide library of the general formula

  11. Scientific opinion on the safety of proline-specific oligopeptidase as a novel food pursuant to Regulation (EC) No 258/97

    DEFF Research Database (Denmark)

    Poulsen, Morten

    2017-01-01

    and of the Council, taking into account the comments and objections of a scientific nature raised by Member States. The novel food is an enzyme preparation of prolyl-oligopeptidase produced with a genetically modified Aspergillus niger self clone strain. The target population is the general adult population......Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver an opinion on proline-specific oligopeptidase (Tolerase® G) as a novel food ingredient submitted pursuant to Regulation (EC) No 258/97 of the European Parliament...

  12. How Are Substrate Binding and Catalysis Affected by Mutating Glu127 and Arg161 in Prolyl-4-hydroxylase? A QM/MM and MD Study

    Science.gov (United States)

    Timmins, Amy; de Visser, Sam P.

    2017-01-01

    Prolyl-4-hydroxylase is a vital enzyme for human physiology involved in the biosynthesis of 4-hydroxyproline, an essential component for collagen formation. The enzyme performs a unique stereo- and regioselective hydroxylation at the C4 position of proline despite the fact that the C5 hydrogen atoms should be thermodynamically easier to abstract. To gain insight into the mechanism and find the origin of this regioselectivity, we have done a quantum mechanics/molecular mechanics (QM/MM) study on wildtype and mutant structures. In a previous study (Timmins et al., 2017) we identified several active site residues critical for substrate binding and positioning. In particular, the Glu127 and Arg161 were shown to form multiple hydrogen bonding and ion-dipole interactions with substrate and could thereby affect the regio- and stereoselectivity of the reaction. In this work, we decided to test that hypothesis and report a QM/MM and molecular dynamics (MD) study on prolyl-4-hydroxylase and several active site mutants where Glu127 or Arg161 are mutated for Asp, Gln, or Lys. Thus, the R161D and R161Q mutants give very high barriers for hydrogen atom abstraction from any proline C–H bond and therefore will be inactive. The R161K mutant, by contrast, sees the regio- and stereoselectivity of the reaction change but still is expected to hydroxylate proline at room temperature. By contrast, the Glu127 mutants E127D and E127Q show possible changes in regioselectivity with the former being more probable to react compared to the latter. PMID:29170737

  13. How Are Substrate Binding and Catalysis Affected by Mutating Glu127 and Arg161 in Prolyl-4-hydroxylase? A QM/MM and MD Study

    Directory of Open Access Journals (Sweden)

    Amy Timmins

    2017-11-01

    Full Text Available Prolyl-4-hydroxylase is a vital enzyme for human physiology involved in the biosynthesis of 4-hydroxyproline, an essential component for collagen formation. The enzyme performs a unique stereo- and regioselective hydroxylation at the C4 position of proline despite the fact that the C5 hydrogen atoms should be thermodynamically easier to abstract. To gain insight into the mechanism and find the origin of this regioselectivity, we have done a quantum mechanics/molecular mechanics (QM/MM study on wildtype and mutant structures. In a previous study (Timmins et al., 2017 we identified several active site residues critical for substrate binding and positioning. In particular, the Glu127 and Arg161 were shown to form multiple hydrogen bonding and ion-dipole interactions with substrate and could thereby affect the regio- and stereoselectivity of the reaction. In this work, we decided to test that hypothesis and report a QM/MM and molecular dynamics (MD study on prolyl-4-hydroxylase and several active site mutants where Glu127 or Arg161 are mutated for Asp, Gln, or Lys. Thus, the R161D and R161Q mutants give very high barriers for hydrogen atom abstraction from any proline C–H bond and therefore will be inactive. The R161K mutant, by contrast, sees the regio- and stereoselectivity of the reaction change but still is expected to hydroxylate proline at room temperature. By contrast, the Glu127 mutants E127D and E127Q show possible changes in regioselectivity with the former being more probable to react compared to the latter.

  14. Prolyl hydroxylation in elastin is not random

    DEFF Research Database (Denmark)

    Schmelzer, Christian E H; Nagel, Marcus B M; Dziomba, Szymon

    2016-01-01

    BACKGROUND: This study aimed to investigate the prolyl and lysine hydroxylation in elastin from different species and tissues. METHODS: Enzymatic digests of elastin samples from human, cattle, pig and chicken were analyzed using mass spectrometry and bioinformatics tools. RESULTS: It was confirmed...... at the protein level that elastin does not contain hydroxylated lysine residues regardless of the species. In contrast, prolyl hydroxylation sites were identified in all elastin samples. Moreover, the analysis of the residues adjacent to prolines allowed the determination of the substrate site preferences...... of prolyl 4-hydroxylase. It was found that elastins from all analyzed species contain hydroxyproline and that at least 20%-24% of all proline residues were partially hydroxylated. Determination of the hydroxylation degrees of specific proline residues revealed that prolyl hydroxylation depends on both...

  15. Hepatitis C virus NS5A protein is a substrate for the peptidyl-prolyl cis/trans isomerase activity of cyclophilins A and B.

    Science.gov (United States)

    Hanoulle, Xavier; Badillo, Aurélie; Wieruszeski, Jean-Michel; Verdegem, Dries; Landrieu, Isabelle; Bartenschlager, Ralf; Penin, François; Lippens, Guy

    2009-05-15

    We report here a biochemical and structural characterization of domain 2 of the nonstructural 5A protein (NS5A) from the JFH1 Hepatitis C virus strain and its interactions with cyclophilins A and B (CypA and CypB). Gel filtration chromatography, circular dichroism spectroscopy, and finally NMR spectroscopy all indicate the natively unfolded nature of this NS5A-D2 domain. Because mutations in this domain have been linked to cyclosporin A resistance, we used NMR spectroscopy to investigate potential interactions between NS5A-D2 and cellular CypA and CypB. We observed a direct molecular interaction between NS5A-D2 and both cyclophilins. The interaction surface on the cyclophilins corresponds to their active site, whereas on NS5A-D2, it proved to be distributed over the many proline residues of the domain. NMR heteronuclear exchange spectroscopy yielded direct evidence that many proline residues in NS5A-D2 form a valid substrate for the enzymatic peptidyl-prolyl cis/trans isomerase (PPIase) activity of CypA and CypB.

  16. Prolyl hydroxylation in elastin is not random.

    Science.gov (United States)

    Schmelzer, Christian E H; Nagel, Marcus B M; Dziomba, Szymon; Merkher, Yulia; Sivan, Sarit S; Heinz, Andrea

    2016-10-01

    This study aimed to investigate the prolyl and lysine hydroxylation in elastin from different species and tissues. Enzymatic digests of elastin samples from human, cattle, pig and chicken were analyzed using mass spectrometry and bioinformatics tools. It was confirmed at the protein level that elastin does not contain hydroxylated lysine residues regardless of the species. In contrast, prolyl hydroxylation sites were identified in all elastin samples. Moreover, the analysis of the residues adjacent to prolines allowed the determination of the substrate site preferences of prolyl 4-hydroxylase. It was found that elastins from all analyzed species contain hydroxyproline and that at least 20%-24% of all proline residues were partially hydroxylated. Determination of the hydroxylation degrees of specific proline residues revealed that prolyl hydroxylation depends on both the species and the tissue, however, is independent of age. The fact that the highest hydroxylation degrees of proline residues were found for elastin from the intervertebral disc and knowledge of elastin arrangement in this tissue suggest that hydroxylation plays a biomechanical role. Interestingly, a proline-rich domain of tropoelastin (domain 24), which contains several repeats of bioactive motifs, does not show any hydroxyproline residues in the mammals studied. The results show that prolyl hydroxylation is not a coincidental feature and may contribute to the adaptation of the properties of elastin to meet the functional requirements of different tissues. The study for the first time shows that prolyl hydroxylation is highly regulated in elastin. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Induced-fit Mechanism for Prolyl Endopeptidase

    Energy Technology Data Exchange (ETDEWEB)

    Li, Min; Chen, Changqing; Davies, David R.; Chiu, Thang K. (NIH); (LSU); (Chinese Aca. Sci.)

    2010-11-15

    Prolyl peptidases cleave proteins at proline residues and are of importance for cancer, neurological function, and type II diabetes. Prolyl endopeptidase (PEP) cleaves neuropeptides and is a drug target for neuropsychiatric diseases such as post-traumatic stress disorder, depression, and schizophrenia. Previous structural analyses showing little differences between native and substrate-bound structures have suggested a lock-and-key catalytic mechanism. We now directly demonstrate from seven structures of Aeromonus punctata PEP that the mechanism is instead induced fit: the native enzyme exists in a conformationally flexible opened state with a large interdomain opening between the {beta}-propeller and {alpha}/{beta}-hydrolase domains; addition of substrate to preformed native crystals induces a large scale conformational change into a closed state with induced-fit adjustments of the active site, and inhibition of this conformational change prevents substrate binding. Absolute sequence conservation among 28 orthologs of residues at the active site and critical residues at the interdomain interface indicates that this mechanism is conserved in all PEPs. This finding has immediate implications for the use of conformationally targeted drug design to improve specificity of inhibition against this family of proline-specific serine proteases.

  18. Identification of conserved prolyl residue important for transport activity and the substrate specificity range of yeast plasma membrane Na(+)/H(+) antiporters

    Czech Academy of Sciences Publication Activity Database

    Zimmermannová, Olga; Zavřel, Martin; Sychrová, Hana

    2005-01-01

    Roč. 280, č. 34 (2005), s. 30638-30647 ISSN 0021-9258 R&D Projects: GA ČR(CZ) GP204/02/D092 Institutional research plan: CEZ:AV0Z5011922 Keywords : yeast * Na+/H+ antiporter * substrate specificity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.854, year: 2005

  19. Structure of the prolyl-tRNA synthetase from the eukaryotic pathogen Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Larson, Eric T.; Kim, Jessica E.; Napuli, Alberto J.; Verlinde, Christophe L. M. J.; Fan, Erkang; Zucker, Frank H.; Van Voorhis, Wesley C.; Buckner, Frederick S.; Hol, Wim G. J.; Merritt, Ethan A., E-mail: merritt@u.washington.edu [Medical Structural Genomics of Pathogenic Protozoa, (United States); University of Washington, Seattle, WA 98195 (United States)

    2012-09-01

    The structure of Giardia prolyl-tRNA synthetase cocrystallized with proline and ATP shows evidence for half-of-the-sites activity, leading to a corresponding mixture of reaction substrates and product (prolyl-AMP) in the two active sites of the dimer. The genome of the human intestinal parasite Giardia lamblia contains only a single aminoacyl-tRNA synthetase gene for each amino acid. The Giardia prolyl-tRNA synthetase gene product was originally misidentified as a dual-specificity Pro/Cys enzyme, in part owing to its unexpectedly high off-target activation of cysteine, but is now believed to be a normal representative of the class of archaeal/eukaryotic prolyl-tRNA synthetases. The 2.2 Å resolution crystal structure of the G. lamblia enzyme presented here is thus the first structure determination of a prolyl-tRNA synthetase from a eukaryote. The relative occupancies of substrate (proline) and product (prolyl-AMP) in the active site are consistent with half-of-the-sites reactivity, as is the observed biphasic thermal denaturation curve for the protein in the presence of proline and MgATP. However, no corresponding induced asymmetry is evident in the structure of the protein. No thermal stabilization is observed in the presence of cysteine and ATP. The implied low affinity for the off-target activation product cysteinyl-AMP suggests that translational fidelity in Giardia is aided by the rapid release of misactivated cysteine.

  20. Prolyl hydroxylase domain enzymes: important regulators of cancer metabolism

    Directory of Open Access Journals (Sweden)

    Yang M

    2014-08-01

    Full Text Available Ming Yang,1 Huizhong Su,1 Tomoyoshi Soga,2 Kamil R Kranc,3 Patrick J Pollard1 1Cancer Biology and Metabolism Group, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK; 2Institute for Advanced Biosciences, Keio University, Mizukami, Tsuruoka, Yamagata, Japan; 3MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK Abstract: The hypoxia-inducible factor (HIF prolyl hydroxylase domain enzymes (PHDs regulate the stability of HIF protein by post-translational hydroxylation of two conserved prolyl residues in its α subunit in an oxygen-dependent manner. Trans-4-prolyl hydroxylation of HIFα under normal oxygen (O2 availability enables its association with the von Hippel-Lindau (VHL tumor suppressor pVHL E3 ligase complex, leading to the degradation of HIFα via the ubiquitin-proteasome pathway. Due to the obligatory requirement of molecular O2 as a co-substrate, the activity of PHDs is inhibited under hypoxic conditions, resulting in stabilized HIFα, which dimerizes with HIFβ and, together with transcriptional co-activators CBP/p300, activates the transcription of its target genes. As a key molecular regulator of adaptive response to hypoxia, HIF plays important roles in multiple cellular processes and its overexpression has been detected in various cancers. The HIF1α isoform in particular has a strong impact on cellular metabolism, most notably by promoting anaerobic, whilst inhibiting O2-dependent, metabolism of glucose. The PHD enzymes also seem to have HIF-independent functions and are subject to regulation by factors other than O2, such as by metabolic status, oxidative stress, and abnormal levels of endogenous metabolites (oncometabolites that have been observed in some types of cancers. In this review, we aim to summarize current understandings of the function and regulation of PHDs in cancer with an emphasis on their roles in metabolism. Keywords: prolyl hydroxylase domain (PHD

  1. Redox modulation of thimet oligopeptidase activity by hydrogen peroxide.

    Science.gov (United States)

    Icimoto, Marcelo Y; Ferreira, Juliana C; Yokomizo, César H; Bim, Larissa V; Marem, Alyne; Gilio, Joyce M; Oliveira, Vitor; Nantes, Iseli L

    2017-07-01

    Thimet oligopeptidase (EC 3.4.24.15, TOP) is a cytosolic mammalian zinc protease that can process a diversity of bioactive peptides. TOP has been pointed out as one of the main postproteasomal enzymes that process peptide antigens in the MHC class I presentation route. In the present study, we describe a fine regulation of TOP activity by hydrogen peroxide (H 2 O 2 ). Cells from a human embryonic kidney cell line (HEK293) underwent an ischemia/reoxygenation-like condition known to increase H 2 O 2 production. Immediately after reoxygenation, HEK293 cells exhibited a 32% increase in TOP activity, but no TOP activity was observed 2 h after reoxygenation. In another model, recombinant rat TOP (rTOP) was challenged by H 2 O 2 produced by rat liver mitoplasts (RLMt) alone, and in combination with antimycin A, succinate, and antimycin A plus succinate. In these conditions, rTOP activity increased 17, 30, 32 and 38%, respectively. Determination of H 2 O 2 concentration generated in reoxygenated cells and mitoplasts suggested a possible modulation of rTOP activity dependent on the concentration of H 2 O 2 . The measure of pure rTOP activity as a function of H 2 O 2 concentration corroborated this hypothesis. The data fitted to an asymmetrical bell-shaped curve in which the optimal activating H 2 O 2 concentration was 1.2 nM, and the maximal inhibition (75% about the control) was 1 μm. Contrary to the oxidation produced by aging associated with enzyme oligomerization and inhibition, H 2 O 2 oxidation produced sulfenic acid and maintained rTOP in the monomeric form. Consistent with the involvement of rTOP in a signaling redox cascade, the H 2 O 2 -oxidized rTOP reacted with dimeric thioredoxin-1 (TRx-1) and remained covalently bound to one subunit of TRx-1.

  2. Characterisation of Aspergillus niger prolyl aminopeptidase

    NARCIS (Netherlands)

    Basten, E.J.W.; Moers, A.P.H.A.; Ooyen, van A.J.J.; Schaap, P.J.

    2005-01-01

    We have cloned a gene (papA) that encodes a prolyl aminopeptidase from Aspergillus niger. Homologous genes are present in the genomes of the Eurotiales A. nidulans, A. fumigatus and Talaromyces emersonii, but the gene is not present in the genome of the yeast Saccharomyces cerevisiae. Cell extracts

  3. Oligopeptidase B from Serratia proteamaculans. III. Inhibition analysis. Specific interactions with metalloproteinase inhibitors.

    Science.gov (United States)

    Mikhailova, A G; Khairullin, R F; Kolomijtseva, G Ya; Rumsh, L D

    2012-03-01

    Inhibition of the novel oligopeptidase B from Serratia proteamaculans (PSP) by basic pancreatic trypsin inhibitor, Zn2+ ions, and o- and m-phenanthroline was investigated. A pronounced effect of calcium ions on the interaction of PSP with inhibitors was demonstrated. Inversion voltamperometry and atomic absorption spectrometry revealed no zinc ions in the PSP molecule. Hydrophobic nature of the enzyme inhibition by o- and m-phenanthroline was established.

  4. Heterologous Expression, Purification and Characterization of an Oligopeptidase A from the Pathogen Leptospira interrogans.

    Science.gov (United States)

    Anu, Prasannan V; Madanan, Madathiparambil G; Nair, Ananthakrishnan J; Nair, Gangaprasad A; Nair, Govinda Pillai M; Sudhakaran, Perumana R; Satheeshkumar, Padikara K

    2018-04-01

    Oligopeptidases are enzymes involved in the degradation of short peptides (generally less than 30 amino acids in size) which help pathogens evade the host defence mechanisms. Leptospira is a zoonotic pathogen and causes leptospirosis in mammals. Proteome analysis of Leptospira revealed the presence of oligopeptidase A (OpdA) among other membrane proteins. To study the role of oligopeptidase in leptospirosis, the OpdA of L. interrogans was cloned and expressed in Escherichia coli with a histidine tag (His-tag). The protein showed maximum expression at 37 °C with 0.5 mM of IPTG after 2 h of induction. Recombinant OpdA protein was purified to homogeneity using Ni-affinity chromatography. The purified OpdA showed more than 80% inhibition with a serine protease inhibitor but the activity was reduced to 30% with the cysteine protease inhibitor. The peptidase activity was increased significantly in the presence of Zn 2+ at a neutral pH. Inhibitor assay indicate the presence of more than one active sites for peptidase activity as reported with the OpdA of E. coli and Salmonella. Over-expression of OpdA in E. coli BL21 (DE3) did not cause any negative effects on normal cell growth and viability. The role of OpdA as virulence factor in Leptospira and its potential as a therapeutic and diagnostic target in leptospirosis is yet to be identified.

  5. Proteolytic activity of Escherichia coli oligopeptidase B against proline-rich antimicrobial peptides

    DEFF Research Database (Denmark)

    Mattiuzzo, Maura; De Gobba, Cristian; Runti, Giulia

    2014-01-01

    Oligopeptidase B (OpdB) is a serine peptidase widespread among bacteria and protozoa that has emerged as a virulence factor despite its function has not yet been precisely established. By using an OpdB-overexpressing Escherichia coli strain, we found that the overexpressed peptidase makes...

  6. Bacillus anthracis Prolyl 4-Hydroxylase Interacts with and Modifies Elongation Factor Tu

    Energy Technology Data Exchange (ETDEWEB)

    Schnicker, Nicholas J. [Department; Razzaghi, Mortezaali [Department; Guha Thakurta, Sanjukta [Department; Chakravarthy, Srinivas [Biophysics; Dey, Mishtu [Department

    2017-10-17

    Prolyl hydroxylation is a very common post-translational modification and plays many roles in eukaryotes such as collagen stabilization, hypoxia sensing, and controlling protein transcription and translation. There is a growing body of evidence that suggests that prokaryotes contain prolyl 4-hydroxylases (P4Hs) homologous to the hypoxia-inducible factor (HIF) prolyl hydroxylase domain (PHD) enzymes that act on elongation factor Tu (EFTu) and are likely involved in the regulation of bacterial translation. Recent biochemical and structural studies with a PHD from Pseudomonas putida (PPHD) determined that it forms a complex with EFTu and hydroxylates a prolyl residue of EFTu. Moreover, while animal, plant, and viral P4Hs act on peptidyl proline, most prokaryotic P4Hs have been known to target free l-proline; the exceptions include PPHD and a P4H from Bacillus anthracis (BaP4H) that modifies collagen-like proline-rich peptides. Here we use biophysical and mass spectrometric methods to demonstrate that BaP4H recognizes full-length BaEFTu and a BaEFTu 9-mer peptide for site-specific proline hydroxylation. Using size-exclusion chromatography coupled small-angle X-ray scattering (SEC–SAXS) and binding studies, we determined that BaP4H forms a 1:1 heterodimeric complex with BaEFTu. The SEC–SAXS studies reveal dissociation of BaP4H dimeric subunits upon interaction with BaEFTu. While BaP4H is unusual within bacteria in that it is structurally and functionally similar to the animal PHDs and collagen P4Hs, respectively, this work provides further evidence of its promiscuous substrate recognition. It is possible that the enzyme might have evolved to hydroxylate a universally conserved protein in prokaryotes, similar to the PHDs, and implies a functional role in B. anthracis.

  7. The Aminolysis Reaction of Streptomyces S9 Aminopeptidase Promotes the Synthesis of Diverse Prolyl Dipeptides▿ †

    Science.gov (United States)

    Arima, Jiro; Morimoto, Masazumi; Usuki, Hirokazu; Mori, Nobuhiro; Hatanaka, Tadashi

    2010-01-01

    Prolyl dipeptide synthesis by S9 aminopeptidase from Streptomyces thermocyaneoviolaceus (S9AP-St) has been demonstrated. In the synthesis, S9AP-St preferentially used l-Pro-OBzl as the acyl donor, yielding synthesized dipeptides having an l-Pro-Xaa structure. In addition, S9AP-St showed broad specificity toward the acyl acceptor. Furthermore, S9AP-St produced cyclo (l-Pro-l-His) with a conversion ratio of substrate to cyclo (l-Pro-l-His) higher than 40%. PMID:20418423

  8. A digestive prolyl carboxypeptidase in Tenebrio molitor larvae.

    Science.gov (United States)

    Goptar, Irina A; Shagin, Dmitry A; Shagina, Irina A; Mudrik, Elena S; Smirnova, Yulia A; Zhuzhikov, Dmitry P; Belozersky, Mikhail A; Dunaevsky, Yakov E; Oppert, Brenda; Filippova, Irina Yu; Elpidina, Elena N

    2013-06-01

    Prolyl carboxypeptidase (PRCP) is a lysosomal proline specific serine peptidase that also plays a vital role in the regulation of physiological processes in mammals. In this report, we isolate and characterize the first PRCP in an insect. PRCP was purified from the anterior midgut of larvae of a stored product pest, Tenebrio molitor, using a three-step chromatography strategy, and it was determined that the purified enzyme was a dimer. The cDNA of PRCP was cloned and sequenced, and the predicted protein was identical to the proteomic sequences of the purified enzyme. The substrate specificity and kinetic parameters of the enzyme were determined. The T. molitor PRCP participates in the hydrolysis of the insect's major dietary proteins, gliadins, and is the first PRCP to be ascribed a digestive function. Our collective data suggest that the evolutionary enrichment of the digestive peptidase complex in insects with an area of acidic to neutral pH in the midgut is a result of the incorporation of lysosomal peptidases, including PRCP. Published by Elsevier Ltd.

  9. Overexpression of hypoxia-inducible factor prolyl- hydoxylase ...

    African Journals Online (AJOL)

    Jane

    2011-08-08

    Aug 8, 2011 ... which is regulated by HIF prolyl-dydoxylase -mediated degradation. Taken together, our results ..... Chromatin immunoprecipitation analysis of gene ... phosphatidylinositol 3-kinase/Akt pathway. Endocrinology, 148(5):.

  10. Cloning, Expression, Sequence Analysis and Homology Modeling of the Prolyl Endoprotease from Eurygaster integriceps Puton

    Directory of Open Access Journals (Sweden)

    Ravi Chandra Yandamuri

    2014-10-01

    Full Text Available eurygaster integriceps Puton, commonly known as sunn pest, is a major pest of wheat in Northern Africa, the Middle East and Eastern Europe. This insect injects a prolyl endoprotease into the wheat, destroying the gluten. The purpose of this study was to clone the full length cDNA of the sunn pest prolyl endoprotease (spPEP for expression in E. coli and to compare the amino acid sequence of the enzyme to other known PEPs in both phylogeny and potential tertiary structure. Sequence analysis shows that the 5ꞌ UTR contains several putative transcription factor binding sites for transcription factors known to be expressed in Drosophila that might be useful targets for inhibition of the enzyme. The spPEP was first identified as a prolyl endoprotease by Darkoh et al., 2010. The enzyme is a unique serine protease of the S9A family by way of its substrate recognition of the gluten proteins, which are greater than 30 kD in size. At 51% maximum identity to known PEPs, homology modeling using SWISS-MODEL, the porcine brain PEP (PDB: 2XWD was selected in the database of known PEP structures, resulting in a predicted tertiary structure 99% identical to the porcine brain PEP structure. A Km for the recombinant spPEP was determined to be 210 ± 53 µM for the zGly-Pro-pNA substrate in 0.025 M ethanolamine, pH 8.5, containing 0.1 M NaCl at 37 °C with a turnover rate of 172 ± 47 µM Gly-Pro-pNA/s/µM of enzyme.

  11. Discovery, cloning and characterisation of proline specific prolyl endopeptidase, a gluten degrading thermo-stable enzyme from Sphaerobacter thermophiles

    DEFF Research Database (Denmark)

    Shetty, Radhakrishna; Vestergaard, Mike; Jessen, Flemming

    2017-01-01

    processes occur at elevated temperature. We present in this paper, the discovery, cloning and characterisation of a novel recombinant thermostable gluten degrading enzyme, a proline specific prolyl endoprotease (PEP) from Sphaerobacter thermophiles. The molecular mass of the prolyl endopeptidase......Gluten free products have emerged during the last decades, as a result of a growing public concern and technological advancements allowing gluten reduction in food products. One approach is to use gluten degrading enzymes, typically at low or ambient temperatures, whereas many food production...... was estimated to be 77 kDa by using SDS-PAGE. Enzyme activity assays with a synthetic dipeptide Z-Gly-Pro-p-nitroanilide as the substrate revealed that the enzyme had optimal activity at pH 6.6 and was most active from pH 5.0-8.0. The optimum temperature was 63 °C and residual activity after one hour incubation...

  12. Structural and Biochemical Characterization of the Human Cyclophilin Family of Peptidyl-Prolyl Isomerases

    Energy Technology Data Exchange (ETDEWEB)

    Davis, Tara L.; Walker, John R.; Campagna-Slater, Valérie; Finerty, Jr., Patrick J.; Paramanathan, Ragika; Bernstein, Galina; MacKenzie, Farrell; Tempel, Wolfram; Ouyang, Hui; Lee, Wen Hwa; Eisenmesser, Elan Z.; Dhe-Paganon, Sirano (Toronto); (Colorado)

    2011-12-14

    Peptidyl-prolyl isomerases catalyze the conversion between cis and trans isomers of proline. The cyclophilin family of peptidyl-prolyl isomerases is well known for being the target of the immunosuppressive drug cyclosporin, used to combat organ transplant rejection. There is great interest in both the substrate specificity of these enzymes and the design of isoform-selective ligands for them. However, the dearth of available data for individual family members inhibits attempts to design drug specificity; additionally, in order to define physiological functions for the cyclophilins, definitive isoform characterization is required. In the current study, enzymatic activity was assayed for 15 of the 17 human cyclophilin isomerase domains, and binding to the cyclosporin scaffold was tested. In order to rationalize the observed isoform diversity, the high-resolution crystallographic structures of seven cyclophilin domains were determined. These models, combined with seven previously solved cyclophilin isoforms, provide the basis for a family-wide structure:function analysis. Detailed structural analysis of the human cyclophilin isomerase explains why cyclophilin activity against short peptides is correlated with an ability to ligate cyclosporin and why certain isoforms are not competent for either activity. In addition, we find that regions of the isomerase domain outside the proline-binding surface impart isoform specificity for both in vivo substrates and drug design. We hypothesize that there is a well-defined molecular surface corresponding to the substrate-binding S2 position that is a site of diversity in the cyclophilin family. Computational simulations of substrate binding in this region support our observations. Our data indicate that unique isoform determinants exist that may be exploited for development of selective ligands and suggest that the currently available small-molecule and peptide-based ligands for this class of enzyme are insufficient for isoform

  13. Targeting Prolyl Peptidases in Triple-Negative Breast Cancer

    Science.gov (United States)

    2017-02-01

    ABSTRACT Triple negative breast cancer (TNBC) is an aggressive sub-type with limited treatment options and poor prognosis. The most life -threatening... negative feedback loops within the pathway limit their effectiveness . For example, AKT inhibitors cause increased expression of IGF1R/ErbB3 and, as a...AWARD NUMBER: W81XWH-16-1-0025 TITLE: Targeting Prolyl Peptidases in Triple- Negative Breast Cancer PRINCIPAL INVESTIGATOR: Carl G. Maki, PhD

  14. Differential effects of collagen prolyl 3-hydroxylation on skeletal tissues.

    Directory of Open Access Journals (Sweden)

    Erica P Homan

    2014-01-01

    Full Text Available Mutations in the genes encoding cartilage associated protein (CRTAP and prolyl 3-hydroxylase 1 (P3H1 encoded by LEPRE1 were the first identified causes of recessive Osteogenesis Imperfecta (OI. These proteins, together with cyclophilin B (encoded by PPIB, form a complex that 3-hydroxylates a single proline residue on the α1(I chain (Pro986 and has cis/trans isomerase (PPIase activity essential for proper collagen folding. Recent data suggest that prolyl 3-hydroxylation of Pro986 is not required for the structural stability of collagen; however, the absence of this post-translational modification may disrupt protein-protein interactions integral for proper collagen folding and lead to collagen over-modification. P3H1 and CRTAP stabilize each other and absence of one results in degradation of the other. Hence, hypomorphic or loss of function mutations of either gene cause loss of the whole complex and its associated functions. The relative contribution of losing this complex's 3-hydroxylation versus PPIase and collagen chaperone activities to the phenotype of recessive OI is unknown. To distinguish between these functions, we generated knock-in mice carrying a single amino acid substitution in the catalytic site of P3h1 (Lepre1(H662A . This substitution abolished P3h1 activity but retained ability to form a complex with Crtap and thus the collagen chaperone function. Knock-in mice showed absence of prolyl 3-hydroxylation at Pro986 of the α1(I and α1(II collagen chains but no significant over-modification at other collagen residues. They were normal in appearance, had no growth defects and normal cartilage growth plate histology but showed decreased trabecular bone mass. This new mouse model recapitulates elements of the bone phenotype of OI but not the cartilage and growth phenotypes caused by loss of the prolyl 3-hydroxylation complex. Our observations suggest differential tissue consequences due to selective inactivation of P3H1 hydroxylase

  15. The basic tilted helix bundle domain of the prolyl isomerase FKBP25 is a novel double-stranded RNA binding module

    Science.gov (United States)

    Dilworth, David; Bonnafous, Pierre; Edoo, Amiirah Bibi; Bourbigot, Sarah; Pesek-Jardim, Francy; Gudavicius, Geoff; Serpa, Jason J.; Petrotchenko, Evgeniy V.; Borchers, Christoph H.

    2017-01-01

    Abstract Prolyl isomerases are defined by a catalytic domain that facilitates the cis–trans interconversion of proline residues. In most cases, additional domains in these enzymes add important biological function, including recruitment to a set of protein substrates. Here, we report that the N-terminal basic tilted helix bundle (BTHB) domain of the human prolyl isomerase FKBP25 confers specific binding to double-stranded RNA (dsRNA). This binding is selective over DNA as well as single-stranded oligonucleotides. We find that FKBP25 RNA-association is required for its nucleolar localization and for the vast majority of its protein interactions, including those with 60S pre-ribosome and early ribosome biogenesis factors. An independent mobility of the BTHB and FKBP catalytic domains supports a model by which the N-terminus of FKBP25 is anchored to regions of dsRNA, whereas the FKBP domain is free to interact with neighboring proteins. Apart from the identification of the BTHB as a new dsRNA-binding module, this domain adds to the growing list of auxiliary functions used by prolyl isomerases to define their primary cellular targets. PMID:29036638

  16. Physiological and Pathogenic Roles of Prolyl Isomerase Pin1 in Metabolic Regulations via Multiple Signal Transduction Pathway Modulations

    Directory of Open Access Journals (Sweden)

    Yusuke Nakatsu

    2016-09-01

    Full Text Available Prolyl isomerases are divided into three groups, the FKBP family, Cyclophilin and the Parvulin family (Pin1 and Par14. Among these isomerases, Pin1 is a unique prolyl isomerase binding to the motif including pSer/pThr-Pro that is phosphorylated by kinases. Once bound, Pin1 modulates the enzymatic activity, protein stability or subcellular localization of target proteins by changing the cis- and trans-formations of proline. Several studies have examined the roles of Pin1 in the pathogenesis of cancers and Alzheimer’s disease. On the other hand, recent studies have newly demonstrated Pin1 to be involved in regulating glucose and lipid metabolism. Interestingly, while Pin1 expression is markedly increased by high-fat diet feeding, Pin1 KO mice are resistant to diet-induced obesity, non-alcoholic steatohepatitis and diabetic vascular dysfunction. These phenomena result from the binding of Pin1 to several key factors regulating metabolic functions, which include insulin receptor substrate-1, AMPK, Crtc2 and NF-κB p65. In this review, we focus on recent advances in elucidating the physiological roles of Pin1 as well as the pathogenesis of disorders involving this isomerase, from the viewpoint of the relationships between signal transductions and metabolic functions.

  17. Production and characterization of two major Aspergillus oryzae secreted prolyl endopeptidases able to efficiently digest proline-rich peptides of gliadin.

    Science.gov (United States)

    Eugster, Philippe J; Salamin, Karine; Grouzmann, Eric; Monod, Michel

    2015-12-01

    Prolyl endopeptidases are key enzymes in the digestion of proline-rich proteins. Fungal extracts rich in prolyl endopeptidases produced by a species such as Aspergillus oryzae used in food fermentation would be of particular interest for the development of an oral enzyme therapy product in patients affected by intolerance to gluten. Two major A. oryzae secreted prolyl endopeptidases of the MEROPS S28 peptidase family, AoS28A and AoS28B, were identified when this fungus was grown at acidic pH in a medium containing soy meal protein or wheat gliadin as the sole source of nitrogen. AoS28B was produced by 12 reference A. oryzae strains used in food fermentation. AoS28A was secreted by six of these 12 strains. This protease is the orthologue of the previously characterized Aspergillus fumigatus (AfuS28) and Aspergillus niger (AN-PEP) prolyl endopeptidases which are encoded by genes with a similar intron-exon structure. Large amounts of secreted AoS28A and AoS28B were obtained by gene overexpression in A. oryzae. AoS28A and AoS28B are endoproteases able to cleave N-terminally blocked proline substrates. Both enzymes very efficiently digested the proline-rich 33-mer of gliadin, the most representative immunotoxic peptide deriving from gliadin, with some differences in terms of specificity and optimal pH. Digestion of the gliadin peptide in short peptides with both enzymes was found to occur from its N terminus.

  18. Sandwich ELISA for quantitative detection of human collagen prolyl 4-hydroxylase

    Directory of Open Access Journals (Sweden)

    Myllyharju Johanna

    2010-06-01

    Full Text Available Abstract Background We describe a method for specific, quantitative and quick detection of human collagen prolyl 4-hydroxylase (C-P4H, the key enzyme for collagen prolyl-4 hydroxylation, in crude samples based on a sandwich ELISA principle. The method is relevant to active C-P4H level monitoring during recombinant C-P4H and collagen production in different expression systems. The assay proves to be specific for the active C-P4H α2β2 tetramer due to the use of antibodies against its both subunits. Thus in keeping with the method C-P4H is captured by coupled to an anti-α subunit antibody magnetic beads and an anti-β subunit antibody binds to the PDI/β subunit of the protein. Then the following holoenzyme detection is accomplished by a goat anti-rabbit IgG labeled with alkaline phosphatase which AP catalyzes the reaction of a substrate transformation with fluorescent signal generation. Results We applied an experimental design approach for the optimization of the antibody concentrations used in the sandwich ELISA. The assay sensitivity was 0.1 ng of C-P4H. The method was utilized for the analysis of C-P4H accumulation in crude cell extracts of E. coli overexpressing C-P4H. The sandwich ELISA signals obtained demonstrated a very good correlation with the detected protein activity levels measured with the standard radioactive assay. The developed assay was applied to optimize C-P4H production in E. coli Origami in a system where the C-P4H subunits expression acted under control by different promoters. The experiments performed in a shake flask fed-batch system (EnBase® verified earlier observations that cell density and oxygen supply are critical factors for the use of the inducer anhydrotetracycline and thus for the soluble C-P4H yield. Conclusions Here we show an example of sandwich ELISA usage for quantifying multimeric proteins. The method was developed for monitoring the amount of recombinant C-P4H tetramer in crude E. coli extracts. Due

  19. Regulation of HIF prolyl hydroxylases by hypoxia-inducible factors.

    Science.gov (United States)

    Aprelikova, Olga; Chandramouli, Gadisetti V R; Wood, Matthew; Vasselli, James R; Riss, Joseph; Maranchie, Jodi K; Linehan, W Marston; Barrett, J Carl

    2004-06-01

    Hypoxia and induction of hypoxia-inducible factors (HIF-1alpha and HIF-2alpha) is a hallmark of many tumors. Under normal oxygen tension HIF-alpha subunits are rapidly degraded through prolyl hydroxylase dependent interaction with the von Hippel-Lindau (VHL) tumor suppressor protein, a component of E3 ubuiquitin ligase complex. Using microarray analysis of VHL mutated and re-introduced cells, we found that one of the prolyl hydroxylases (PHD3) is coordinately expressed with known HIF target genes, while the other two family members (PHD1 and 2) did not respond to VHL. We further tested the regulation of these genes by HIF-1 and HIF-2 and found that siRNA targeted degradation of HIF-1alpha and HIF-2alpha results in decreased hypoxia-induced PHD3 expression. Ectopic overexpression of HIF-2alpha in two different cell lines provided a much better induction of PHD3 gene than HIF-1alpha. In contrast, we demonstrate that PHD2 is not affected by overexpression or downregulation of HIF-2alpha. However, induction of PHD2 by hypoxia has HIF-1-independent and -dependent components. Short-term hypoxia (4 h) results in induction of PHD2 independent of HIF-1, while PHD2 accumulation by prolonged hypoxia (16 h) was decreased by siRNA-mediated degradation of HIF-1alpha subunit. These data further advance our understanding of the differential role of HIF factors and putative feedback loop in HIF regulation. Copyright 2004 Wiley-Liss, Inc.

  20. Prolyl 3-hydroxylase 1 and CRTAP are mutually stabilizing in the endoplasmic reticulum collagen prolyl 3-hydroxylation complex.

    Science.gov (United States)

    Chang, Weizhong; Barnes, Aileen M; Cabral, Wayne A; Bodurtha, Joann N; Marini, Joan C

    2010-01-15

    Null mutations in cartilage-associated protein (CRTAP) and prolyl 3-hydroxylase 1 (P3H1/LEPRE1) cause types VII and VIII OI, respectively, two novel recessive forms of osteogenesis imperfecta (OI) with severe to lethal bone dysplasia and overmodification of the type I collagen helical region. CRTAP and P3H1 form a complex with cyclophilin B (CyPB) in the endoplasmic reticulum (ER) which 3-hydroxylates the Pro986 residue of alpha1(I) and alpha1(II) collagen chains. We investigated the interaction of complex components in fibroblasts from types VII and VIII OI patients. Both CRTAP and P3H1 are absent or reduced on western blots and by immunofluorescence microscopy in cells containing null mutations in either gene. Levels of LEPRE1 or CRTAP transcripts, however, are normal in CRTAP- or LEPRE1-null cells, respectively. Stable transfection of a CRTAP or LEPRE1 expression construct into cells with null mutations for the transfected cDNA restored both CRTAP and P3H1 protein levels. Normalization of collagen helical modification in transfected CRTAP-null cells demonstrated that the restored proteins functioned effectively as a complex. These data indicate that CRTAP and P3H1 are mutually stabilized in the collagen prolyl 3-hydroxylation complex. CyPB levels were unaffected by mutations in either CRTAP or LEPRE1. Proteasomal inhibitors partially rescue P3H1 protein in CRTAP-null cells. In LEPRE1-null cells, secretion of CRTAP is increased compared with control cells and accounts for 15-20% of the decreased CRTAP detected in cells. Thus, mutual stabilization of P3H1 and CRTAP in the ER collagen modification complex is an underlying mechanism for the overlapping phenotype of types VII and VIII OI.

  1. Roles of Prolyl Isomerases in RNA-Mediated Gene Expression

    Directory of Open Access Journals (Sweden)

    Roopa Thapar

    2015-05-01

    Full Text Available The peptidyl-prolyl cis-trans isomerases (PPIases that include immunophilins (cyclophilins and FKBPs and parvulins (Pin1, Par14, Par17 participate in cell signaling, transcription, pre-mRNA processing and mRNA decay. The human genome encodes 19 cyclophilins, 18 FKBPs and three parvulins. Immunophilins are receptors for the immunosuppressive drugs cyclosporin A, FK506, and rapamycin that are used in organ transplantation. Pin1 has also been targeted in the treatment of Alzheimer’s disease, asthma, and a number of cancers. While these PPIases are characterized as molecular chaperones, they also act in a nonchaperone manner to promote protein-protein interactions using surfaces outside their active sites. The immunosuppressive drugs act by a gain-of-function mechanism by promoting protein-protein interactions in vivo. Several immunophilins have been identified as components of the spliceosome and are essential for alternative splicing. Pin1 plays roles in transcription and RNA processing by catalyzing conformational changes in the RNA Pol II C-terminal domain. Pin1 also binds several RNA binding proteins such as AUF1, KSRP, HuR, and SLBP that regulate mRNA decay by remodeling mRNP complexes. The functions of ribonucleoprotein associated PPIases are largely unknown. This review highlights PPIases that play roles in RNA-mediated gene expression, providing insight into their structures, functions and mechanisms of action in mRNP remodeling in vivo.

  2. cDNA cloning of porcine brain prolyl endopeptidase and identification of the active-site seryl residue

    Energy Technology Data Exchange (ETDEWEB)

    Rennex, D.; Hemmings, B.A.; Hofsteenge, J.; Stone, S.R. (Friedrich Miescher-Institut, Basel (Switzerland))

    1991-02-26

    Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. ({sup 3}H)Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease.

  3. Biochemical characterization of the prolyl 3-hydroxylase 1.cartilage-associated protein.cyclophilin B complex.

    Science.gov (United States)

    Ishikawa, Yoshihiro; Wirz, Jackie; Vranka, Janice A; Nagata, Kazuhiro; Bächinger, Hans Peter

    2009-06-26

    The rough endoplasmic reticulum-resident protein complex consisting of prolyl 3-hydroxylase 1 (P3H1), cartilage-associated protein (CRTAP), and cyclophilin B (CypB) can be isolated from chick embryos on a gelatin-Sepharose column, indicating some involvement in the biosynthesis of procollagens. Prolyl 3-hydroxylase 1 modifies a single proline residue in the alpha chains of type I, II, and III collagens to (3S)-hydroxyproline. The peptidyl-prolyl cis-trans isomerase activity of cyclophilin B was shown previously to catalyze the rate of triple helix formation. Here we show that cyclophilin B in the complex shows peptidyl-prolyl cis-trans isomerase activity and that the P3H1.CRTAP.CypB complex has another important function: it acts as a chaperone molecule when tested with two classical chaperone assays. The P3H1.CRTAP.CypB complex inhibited the thermal aggregation of citrate synthase and was active in the denatured rhodanese refolding and aggregation assay. The chaperone activity of the complex was higher than that of protein-disulfide isomerase, a well characterized chaperone. The P3H1.CRTAP.CypB complex also delayed the in vitro fibril formation of type I collagen, indicating that this complex is also able to interact with triple helical collagen and acts as a collagen chaperone.

  4. Inhibiting prolyl isomerase activity by hybrid organic-inorganic molecules containing rhodium(II) fragments.

    Science.gov (United States)

    Coughlin, Jane M; Kundu, Rituparna; Cooper, Julian C; Ball, Zachary T

    2014-11-15

    A small molecule containing a rhodium(II) tetracarboxylate fragment is shown to be a potent inhibitor of the prolyl isomerase FKBP12. The use of small molecules conjugates of rhodium(II) is presented as a general strategy for developing new protein inhibitors based on distinct structural and sequence features of the enzyme active site. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Interaction of p53 with prolyl isomerases: Healthy and unhealthy relationships.

    Science.gov (United States)

    Mantovani, Fiamma; Zannini, Alessandro; Rustighi, Alessandra; Del Sal, Giannino

    2015-10-01

    The p53 protein family, comprising p53, p63 and p73, is primarily involved in preserving genome integrity and preventing tumor onset, and also affects a range of physiological processes. Signal-dependent modifications of its members and of other pathway components provide cells with a sophisticated code to transduce a variety of stress signaling into appropriate responses. TP53 mutations are highly frequent in cancer and lead to the expression of mutant p53 proteins that are endowed with oncogenic activities and sensitive to stress signaling. p53 family proteins have unique structural and functional plasticity, and here we discuss the relevance of prolyl-isomerization to actively shape these features. The anti-proliferative functions of the p53 family are carefully activated upon severe stress and this involves the interaction with prolyl-isomerases. In particular, stress-induced stabilization of p53, activation of its transcriptional control over arrest- and cell death-related target genes and of its mitochondrial apoptotic function, as well as certain p63 and p73 functions, all require phosphorylation of specific S/T-P motifs and their subsequent isomerization by the prolyl-isomerase Pin1. While these functions of p53 counteract tumorigenesis, under some circumstances their activation by prolyl-isomerases may have negative repercussions (e.g. tissue damage induced by anticancer therapies and ischemia-reperfusion, neurodegeneration). Moreover, elevated Pin1 levels in tumor cells may transduce deregulated phosphorylation signaling into activation of mutant p53 oncogenic functions. The complex repertoire of biological outcomes induced by p53 finds mechanistic explanations, at least in part, in the association between prolyl-isomerases and the p53 pathway. This article is part of a Special Issue entitled Proline-directed foldases: Cell signaling catalysts and drug targets. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Peptidyl prolyl isomerase Pin1-inhibitory activity of D-glutamic and D-aspartic acid derivatives bearing a cyclic aliphatic amine moiety.

    Science.gov (United States)

    Nakagawa, Hidehiko; Seike, Suguru; Sugimoto, Masatoshi; Ieda, Naoya; Kawaguchi, Mitsuyasu; Suzuki, Takayoshi; Miyata, Naoki

    2015-12-01

    Pin1 is a peptidyl prolyl isomerase that specifically catalyzes cis-trans isomerization of phosphorylated Thr/Ser-Pro peptide bonds in substrate proteins and peptides. Pin1 is involved in many important cellular processes, including cancer progression, so it is a potential target of cancer therapy. We designed and synthesized a novel series of Pin1 inhibitors based on a glutamic acid or aspartic acid scaffold bearing an aromatic moiety to provide a hydrophobic surface and a cyclic aliphatic amine moiety with affinity for the proline-binding site of Pin1. Glutamic acid derivatives bearing cycloalkylamino and phenylthiazole groups showed potent Pin1-inhibitory activity comparable with that of known inhibitor VER-1. The results indicate that steric interaction of the cyclic alkyl amine moiety with binding site residues plays a key role in enhancing Pin1-inhibitory activity. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. Potent and Selective Triazole-Based Inhibitors of the Hypoxia-Inducible Factor Prolyl-Hydroxylases with Activity in the Murine Brain.

    Directory of Open Access Journals (Sweden)

    Mun Chiang Chan

    Full Text Available As part of the cellular adaptation to limiting oxygen availability in animals, the expression of a large set of genes is activated by the upregulation of the hypoxia-inducible transcription factors (HIFs. Therapeutic activation of the natural human hypoxic response can be achieved by the inhibition of the hypoxia sensors for the HIF system, i.e. the HIF prolyl-hydroxylases (PHDs. Here, we report studies on tricyclic triazole-containing compounds as potent and selective PHD inhibitors which compete with the 2-oxoglutarate co-substrate. One compound (IOX4 induces HIFα in cells and in wildtype mice with marked induction in the brain tissue, revealing that it is useful for studies aimed at validating the upregulation of HIF for treatment of cerebral diseases including stroke.

  8. Negative Regulation of the Stability and Tumor Suppressor Function of Fbw7 by the Pin1 Prolyl Isomerase

    Science.gov (United States)

    Min, Sang-Hyun; Lau, Alan W.; Lee, Tae Ho; Inuzuka, Hiroyuki; Wei, Shuo; Huang, Pengyu; Shaik, Shavali; Lee, Daniel Yenhong; Finn, Greg; Balastik, Martin; Chen, Chun-Hau; Luo, Manli; Tron, Adriana E.; DeCaprio, James A.; Zhou, Xiao Zhen; Wei, Wenyi; Lu, Kun Ping

    2012-01-01

    SUMMARY Fbw7 is the substrate recognition component of the SCF (Skp1-Cullin-F-box)-type E3 ligase complex and a well-characterized tumor suppressor that targets numerous oncoproteins for destruction. Genomic deletion or mutation of FBW7 has been frequently found in various types of human cancers, however, little is known about the upstream signaling pathway(s) governing Fbw7 stability and cellular functions. Here we report that Fbw7 protein destruction and tumor suppressor function are negatively regulated by the prolyl isomerase Pin1. Pin1 interacts with Fbw7 in a phoshorylation-dependent manner and promotes Fbw7 self-ubiquitination and protein degradation by disrupting Fbw7 dimerization. Consequently, over-expressing Pin1 reduces Fbw7 abundance and suppresses Fbw7’s ability to inhibit proliferation and transformation. By contrast, depletion of Pin1 in cancer cells leads to elevated Fbw7 expression, which subsequently reduces Mcl-1 abundance, sensitizing cancer cells to Taxol. Thus, Pin1-mediated inhibition of Fbw7 contributes to oncogenesis and Pin1 may be a promising drug target for anti-cancer therapy. PMID:22608923

  9. X-prolyl dipeptidyl aminopeptidase gene (pepX) is part of the glnRA operon in Lactobacillus rhamnosus.

    Science.gov (United States)

    Varmanen, P; Savijoki, K; Avall, S; Palva, A; Tynkkynen, S

    2000-01-01

    A peptidase gene expressing X-prolyl dipeptidyl aminopeptidase (PepX) activity was cloned from Lactobacillus rhamnosus 1/6 by using the chromogenic substrate L-glycyl-L-prolyl-beta-naphthylamide for screening of a genomic library in Escherichia coli. The nucleotide sequence of a 3.5-kb HindIII fragment expressing the peptidase activity revealed one complete open reading frame (ORF) of 2,391 nucleotides. The 797-amino-acid protein encoded by this ORF was shown to be 40, 39, and 36% identical with PepXs from Lactobacillus helveticus, Lactobacillus delbrueckii, and Lactococcus lactis, respectively. By Northern analysis with a pepX-specific probe, transcripts of 4.5 and 7.0 kb were detected, indicating that pepX is part of a polycistronic operon in L. rhamnosus. Cloning and sequencing of the upstream region of pepX revealed the presence of two ORFs of 360 and 1,338 bp that were shown to be able to encode proteins with high homology to GlnR and GlnA proteins, respectively. By multiple primer extension analyses, the only functional promoter in the pepX region was located 25 nucleotides upstream of glnR. Northern analysis with glnA- and pepX-specific probes indicated that transcription from glnR promoter results in a 2.0-kb dicistronic glnR-glnA transcript and also in a longer read-through polycistronic transcript of 7.0 kb that was detected with both probes in samples from cells in exponential growth phase. The glnA gene was disrupted by a single-crossover recombinant event using a nonreplicative plasmid carrying an internal part of glnA. In the disruption mutant, glnRA-specific transcription was derepressed 10-fold compared to the wild type, but the 7.0-kb transcript was no longer detectable with either the glnA- or pepX-specific probe, demonstrating that pepX is indeed part of glnRA operon in L. rhamnosus. Reverse transcription-PCR analysis further supported this operon structure. An extended stem-loop structure was identified immediately upstream of pepX in the gln

  10. Crystallization and preliminary X-ray characterization of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis

    International Nuclear Information System (INIS)

    Nakajima, Yoshitaka; Ito, Kiyoshi; Xu, Yue; Yamada, Nozomi; Onohara, Yuko; Ito, Takashi; Yoshimoto, Tadashi

    2005-01-01

    P. gingivalis prolyl tripeptidyl aminopeptidase has been crystallized by the vapour-diffusion method. Diffraction data have been collected and processed to 2.1 Å resolution. A recombinant form of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis has been crystallized by the hanging-drop vapour-diffusion method using potassium sodium tartrate as a precipitating agent. The crystals belong to the hexagonal space group P6 3 22, with unit-cell parameters a = b = 149.4, c = 159.7 Å. The crystals are most likely to contain one subunit of a dimer in the asymmetric unit, with a V M value of 3.14 Å 3 Da −1 . Diffraction data were collected to 2.1 Å resolution using synchrotron radiation at the BL5 station of the Photon Factory

  11. Crystallization and preliminary X-ray characterization of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Nakajima, Yoshitaka; Ito, Kiyoshi; Xu, Yue; Yamada, Nozomi; Onohara, Yuko; Ito, Takashi; Yoshimoto, Tadashi [Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8521 (Japan)

    2005-12-01

    P. gingivalis prolyl tripeptidyl aminopeptidase has been crystallized by the vapour-diffusion method. Diffraction data have been collected and processed to 2.1 Å resolution. A recombinant form of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis has been crystallized by the hanging-drop vapour-diffusion method using potassium sodium tartrate as a precipitating agent. The crystals belong to the hexagonal space group P6{sub 3}22, with unit-cell parameters a = b = 149.4, c = 159.7 Å. The crystals are most likely to contain one subunit of a dimer in the asymmetric unit, with a V{sub M} value of 3.14 Å{sup 3} Da{sup −1}. Diffraction data were collected to 2.1 Å resolution using synchrotron radiation at the BL5 station of the Photon Factory.

  12. A sandwich immunoassay for human prolyl 4-hydroxylase using monoclonal antibody

    International Nuclear Information System (INIS)

    Yoshida, Shinichi

    1986-01-01

    Monoclonal antibody was used in a sandwich enzyme immunoassay and in a radioimmunoassay for human serum immunoreactive prolyl 4-hydroxylase. The enzyme immunoassay utilized a monoclonal antibody as a solid phase and horseradish peroxidase-labeled rabbit antibody to human prolyl 4-hydroxylase as a conjugate. Sensitivity was 0.1 ng of enzyme per tube. With a conjugate purified by an enzyme-bound affinity column, sensitivity was increased to 0.01 ng per tube, and linearity was obtained between 0.01 to 30 ng per tube. The radioimmunoassay used a 125 I-labeled rabbit antibody (IgG) as the conjugate. Sensitivity of this technique was 0.4 ng of enzyme per tube. (Auth.)

  13. Prolyl hydroxylation regulates protein degradation, synthesis, and splicing in human induced pluripotent stem cell-derived cardiomyocytes.

    Science.gov (United States)

    Stoehr, Andrea; Yang, Yanqin; Patel, Sajni; Evangelista, Alicia M; Aponte, Angel; Wang, Guanghui; Liu, Poching; Boylston, Jennifer; Kloner, Philip H; Lin, Yongshun; Gucek, Marjan; Zhu, Jun; Murphy, Elizabeth

    2016-06-01

    Protein hydroxylases are oxygen- and α-ketoglutarate-dependent enzymes that catalyse hydroxylation of amino acids such as proline, thus linking oxygen and metabolism to enzymatic activity. Prolyl hydroxylation is a dynamic post-translational modification that regulates protein stability and protein-protein interactions; however, the extent of this modification is largely uncharacterized. The goals of this study are to investigate the biological consequences of prolyl hydroxylation and to identify new targets that undergo prolyl hydroxylation in human cardiomyocytes. We used human induced pluripotent stem cell-derived cardiomyocytes in combination with pulse-chase amino acid labelling and proteomics to analyse the effects of prolyl hydroxylation on protein degradation and synthesis. We identified 167 proteins that exhibit differences in degradation with inhibition of prolyl hydroxylation by dimethyloxalylglycine (DMOG); 164 were stabilized. Proteins involved in RNA splicing such as serine/arginine-rich splicing factor 2 (SRSF2) and splicing factor and proline- and glutamine-rich (SFPQ) were stabilized with DMOG. DMOG also decreased protein translation of cytoskeletal and sarcomeric proteins such as α-cardiac actin. We searched the mass spectrometry data for proline hydroxylation and identified 134 high confidence peptides mapping to 78 unique proteins. We identified SRSF2, SFPQ, α-cardiac actin, and cardiac titin as prolyl hydroxylated. We identified 29 prolyl hydroxylated proteins that showed a significant difference in either protein degradation or synthesis. Additionally, we performed next-generation RNA sequencing and showed that the observed decrease in protein synthesis was not due to changes in mRNA levels. Because RNA splicing factors were prolyl hydroxylated, we investigated splicing ± inhibition of prolyl hydroxylation and detected 369 alternative splicing events, with a preponderance of exon skipping. This study provides the first extensive

  14. Theileria parasites secrete a prolyl isomerase to maintain host leukocyte transformation.

    Science.gov (United States)

    Marsolier, J; Perichon, M; DeBarry, J D; Villoutreix, B O; Chluba, J; Lopez, T; Garrido, C; Zhou, X Z; Lu, K P; Fritsch, L; Ait-Si-Ali, S; Mhadhbi, M; Medjkane, S; Weitzman, J B

    2015-04-16

    Infectious agents develop intricate mechanisms to interact with host cell pathways and hijack their genetic and epigenetic machinery to change host cell phenotypic states. Among the Apicomplexa phylum of obligate intracellular parasites, which cause veterinary and human diseases, Theileria is the only genus that transforms its mammalian host cells. Theileria infection of bovine leukocytes induces proliferative and invasive phenotypes associated with activated signalling pathways, notably JNK and AP-1 (ref. 2). The transformed phenotypes are reversed by treatment with the theilericidal drug buparvaquone. We used comparative genomics to identify a homologue of the peptidyl-prolyl isomerase PIN1 in T. annulata (TaPIN1) that is secreted into the host cell and modulates oncogenic signalling pathways. Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation. We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain. Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

  15. Cellular Oxygen Sensing: Crystal Structure of Hypoxia-Inducible Factor Prolyl Hydroxylase (PHD2)

    Energy Technology Data Exchange (ETDEWEB)

    McDonough,M.; Li, V.; Flashman, E.; Chowdhury, R.; Mohr, C.; Lienard, B.; Zondlo, J.; Oldham, N.; Clifton, I.; et al.

    2006-01-01

    Cellular and physiological responses to changes in dioxygen levels in metazoans are mediated via the posttranslational oxidation of hypoxia-inducible transcription factor (HIF). Hydroxylation of conserved prolyl residues in the HIF-{alpha} subunit, catalyzed by HIF prolyl-hydroxylases (PHDs), signals for its proteasomal degradation. The requirement of the PHDs for dioxygen links changes in dioxygen levels with the transcriptional regulation of the gene array that enables the cellular response to chronic hypoxia; the PHDs thus act as an oxygen-sensing component of the HIF system, and their inhibition mimics the hypoxic response. We describe crystal structures of the catalytic domain of human PHD2, an important prolyl-4-hydroxylase in the human hypoxic response in normal cells, in complex with Fe(II) and an inhibitor to 1.7 Angstroms resolution. PHD2 crystallizes as a homotrimer and contains a double-stranded {beta}-helix core fold common to the Fe(II) and 2-oxoglutarate-dependant dioxygenase family, the residues of which are well conserved in the three human PHD enzymes (PHD 1-3). The structure provides insights into the hypoxic response, helps to rationalize a clinically observed mutation leading to familial erythrocytosis, and will aid in the design of PHD selective inhibitors for the treatment of anemia and ischemic disease.

  16. Complex Regulation of Prolyl-4-Hydroxylases Impacts Root Hair Expansion

    DEFF Research Database (Denmark)

    Velasquez, Silvia M; Ricardi, Martiniano M; Poulsen, Christian Peter

    2015-01-01

    Root hairs are single cells that develop by tip growth, a process shared with pollen tubes, axons, and fungal hyphae. However, structural plant cell walls impose constraints to accomplish tip growth. In addition to polysaccharides, plant cell walls are composed of hydroxyproline-rich glycoproteins......5, and to a lesser extent P4H2 and P4H13, are pivotal for root hair tip growth. Second, we demonstrate that P4H5 has in vitro preferred specificity for EXT substrates rather than for other HRGPs. Third, by P4H promoter and protein swapping approaches, we show that P4H2 and P4H13 have interchangeable...... peptidyl-proline hydroxylation on EXTs, and possibly in other HRGPs, is required for proper cell wall self-assembly and hence root hair elongation in Arabidopsis thaliana....

  17. Regulation of Axonal Midline Guidance by Prolyl 4-Hydroxylation in Caenorhabditis elegans

    DEFF Research Database (Denmark)

    Torpe, Nanna; Pocock, Roger David John

    2014-01-01

    , little is known of its importance in the control of axon guidance. In a screen of prolyl 4-hydroxylase (P4H) mutants, we found that genetic removal of a specific P4H subunit, DPY-18, causes dramatic defects in C. elegans neuroanatomy. In dpy-18 mutant animals, the axons of specific ventral nerve cord......Neuronal wiring during development requires that the growth cones of axons and dendrites are correctly guided to their appropriate targets. As in other animals, axon growth cones in Caenorhabditis elegans integrate information in their extracellular environment via interactions among transiently...

  18. Biochemical Characterization of the Prolyl 3-Hydroxylase 1·Cartilage-associated Protein·Cyclophilin B Complex*

    Science.gov (United States)

    Ishikawa, Yoshihiro; Wirz, Jackie; Vranka, Janice A.; Nagata, Kazuhiro; Bächinger, Hans Peter

    2009-01-01

    The rough endoplasmic reticulum-resident protein complex consisting of prolyl 3-hydroxylase 1 (P3H1), cartilage-associated protein (CRTAP), and cyclophilin B (CypB) can be isolated from chick embryos on a gelatin-Sepharose column, indicating some involvement in the biosynthesis of procollagens. Prolyl 3-hydroxylase 1 modifies a single proline residue in the α chains of type I, II, and III collagens to (3S)-hydroxyproline. The peptidyl-prolyl cis-trans isomerase activity of cyclophilin B was shown previously to catalyze the rate of triple helix formation. Here we show that cyclophilin B in the complex shows peptidyl-prolyl cis-trans isomerase activity and that the P3H1·CRTAP·CypB complex has another important function: it acts as a chaperone molecule when tested with two classical chaperone assays. The P3H1·CRTAP·CypB complex inhibited the thermal aggregation of citrate synthase and was active in the denatured rhodanese refolding and aggregation assay. The chaperone activity of the complex was higher than that of protein-disulfide isomerase, a well characterized chaperone. The P3H1·CRTAP·CypB complex also delayed the in vitro fibril formation of type I collagen, indicating that this complex is also able to interact with triple helical collagen and acts as a collagen chaperone. PMID:19419969

  19. Recent Advances in Developing Inhibitors for Hypoxia-Inducible Factor Prolyl Hydroxylases and Their Therapeutic Implications

    Directory of Open Access Journals (Sweden)

    So Yeon Kim

    2015-11-01

    Full Text Available Hypoxia-inducible factor (HIF prolyl hydroxylases (PHDs are members of the 2-oxoglutarate dependent non-heme iron dioxygenases. Due to their physiological roles in regulation of HIF-1α stability, many efforts have been focused on searching for selective PHD inhibitors to control HIF-1α levels for therapeutic applications. In this review, we first describe the structure of PHD2 as a molecular basis for structure-based drug design (SBDD and various experimental methods developed for measuring PHD activity. We further discuss the current status of the development of PHD inhibitors enabled by combining SBDD approaches with high-throughput screening. Finally, we highlight the clinical implications of small molecule PHD inhibitors.

  20. Effects of peptidyl-prolyl isomerase 1 depletion in animal models of prion diseases.

    Science.gov (United States)

    Legname, Giuseppe; Virgilio, Tommaso; Bistaffa, Edoardo; De Luca, Chiara Maria Giulia; Catania, Marcella; Zago, Paola; Isopi, Elisa; Campagnani, Ilaria; Tagliavini, Fabrizio; Giaccone, Giorgio; Moda, Fabio

    2018-04-20

    Pin1 is a peptidyl-prolyl isomerase that induces the cis-trans conversion of specific Ser/Thr-Pro peptide bonds in phosphorylated proteins, leading to conformational changes through which Pin1 regulates protein stability and activity. Since down-regulation of Pin1 has been described in several neurodegenerative disorders, including Alzheimer's Disease (AD), Parkinson's Disease (PD) and Huntington's Disease (HD), we investigated its potential role in prion diseases. Animals generated on wild-type (Pin1 +/+ ), hemizygous (Pin1 +/- ) or knock-out (Pin1 -/- ) background for Pin1 were experimentally infected with RML prions. The study indicates that, neither the total depletion nor reduced levels of Pin1 significantly altered the clinical and neuropathological features of the disease.

  1. Helicobacter pylori Peptidyl Prolyl Isomerase Expression Is Associated with the Severity of Gastritis.

    Science.gov (United States)

    Oghalaie, Akbar; Saberi, Samaneh; Esmaeili, Maryam; Ebrahimzadeh, Fatemeh; Barkhordari, Farzaneh; Ghamarian, Abdolreza; Tashakoripoor, Mohammad; Abdirad, Afshin; Eshagh Hosseini, Mahmoud; Khalaj, Vahid; Mohammadi, Marjan

    2016-12-01

    Helicobacter pylori secretory peptidyl prolyl isomerase, HP0175, is progressively identified as a pro-inflammatory and pro-carcinogenic protein, which serves to link H. pylori infection to its more severe clinical outcomes. Here, we have analyzed host HP0175-specific antibody responses in relation to the severity of gastritis. The HP0175 gene fragment was PCR-amplified, cloned, expressed and purified by Ni-NTA affinity chromatography. Serum antigen-specific antibody responses of non-ulcer dyspeptic patients (N = 176) against recombinant HP0175 were detected by western blotting. The infection status of these subjects was determined by rapid urease test, culture, histology, and serology. The grade of inflammation and stage of atrophy were scored blindly according to the OLGA staging system. The recombinant HP0175 (rHP0175) was expressed as a ~35 kDa protein and its identity was confirmed by western blotting using anti-6X His tag antibody and pooled H. pylori-positive sera. Serum IgG antibodies against rHP0175 segregated our patients into two similar-sized groups of sero-positives (90/176, 51.1 %) and sero-negatives (86/176, 48.9 %). The former presented with higher grades of gastric inflammation (OR = 4.4, 95 % CI = 1.9-9.9, P = 0.001) and stages of gastric atrophy (OR = 18.3, 95 %CI = 1.4-246.6, P = 0.028). Our findings lend further support to the pro-inflammatory nature of H. pylori peptidyl prolyl isomerase (HP0175) and recommends this antigen as a non-invasive serum biomarker of the severity of H. pylori-associated gastritis.

  2. Optimized expression of prolyl aminopeptidase in Pichia pastoris and its characteristics after glycosylation.

    Science.gov (United States)

    Yang, Hongyu; Zhu, Qiang; Zhou, Nandi; Tian, Yaping

    2016-11-01

    Prolyl aminopeptidases are specific exopeptidases that catalyze the hydrolysis of the N-terminus proline residue of peptides and proteins. In the present study, the prolyl aminopeptidase gene (pap) from Aspergillus oryzae JN-412 was optimized through the codon usage of Pichia pastoris. Both the native and optimized pap genes were inserted into the expression vector pPIC9 K and were successfully expressed in P. pastoris. Additionally, the activity of the intracellular enzyme expressed by the recombinant optimized pap gene reached 61.26 U mL(-1), an activity that is 2.1-fold higher than that of the native gene. The recombinant enzyme was purified by one-step elution through Ni-affinity chromatography. The optimal temperature and pH of the purified PAP were 60 °C and 7.5, respectively. Additionally, the recombinant PAP was recovered at a yield greater than 65 % at an extremely broad range of pH values from 6 to 10 after treatment at 50 °C for 6 h. The molecular weight of the recombinant PAP decreased from 50 kDa to 48 kDa after treatment with a deglycosylation enzyme, indicating that the recombinant PAP was completely glycosylated. The glycosylated PAP exhibited high thermo-stability. Half of the activity remained after incubation at 50 °C for 50 h, whereas the remaining activity of PAP expressed in E. coli was only 10 % after incubation at 50 °C for 1 h. PAP could be activated by the appropriate salt concentration and exhibited salt tolerance against NaCl at a concentration up to 5 mol L(-1).

  3. Prolyl carboxypeptidase in Agouti-related Peptide neurons modulates food intake and body weight

    Directory of Open Access Journals (Sweden)

    Giuseppe Bruschetta

    2018-04-01

    Full Text Available Objective: Prolyl carboxypeptidase (PRCP plays a role in the regulation of energy metabolism by inactivating hypothalamic α-melanocyte stimulating hormone (α-MSH levels. Although detected in the arcuate nucleus, limited PRCP expression has been observed in the arcuate POMC neurons, and its site of action in regulating metabolism is still ill-defined. Methods: We performed immunostaining to assess the localization of PRCP in arcuate Neuropeptide Y/Agouti-related Peptide (NPY/AgRP neurons. Hypothalamic explants were then used to assess the intracellular localization of PRCP and its release at the synaptic levels. Finally, we generated a mouse model to assess the role of PRCP in NPY/AgRP neurons of the arcuate nucleus in the regulation of metabolism. Results: Here we show that PRCP is expressed in NPY/AgRP-expressing neurons of the arcuate nucleus. In hypothalamic explants, stimulation by ghrelin increased PRCP concentration in the medium and decreased PRCP content in synaptic extract, suggesting that PRCP is released at the synaptic level. In support of this, hypothalamic explants from mice with selective deletion of PRCP in AgRP neurons (PrcpAgRPKO showed reduced ghrelin-induced PRCP concentration in the medium compared to controls mice. Furthermore, male PrcpAgRPKO mice had decreased body weight and fat mass compared to controls. However, this phenotype was sex-specific as female PrcpAgRPKO mice show metabolic differences only when challenged by high fat diet feeding. The improved metabolism of PrcpAgRPKO mice was associated with reduced food intake and increased energy expenditure, locomotor activity, and hypothalamic α-MSH levels. Administration of SHU9119, a potent melanocortin receptor antagonist, selectively in the PVN of PrcpAgRPKO male mice increased food intake to a level similar to that of control mice. Conclusions: Altogether, our data indicate that PRCP is released at the synaptic levels and that PRCP in AgRP neurons contributes to

  4. Radical scavenging, prolyl endopeptidase inhibitory, and antimicrobial potential of a cultured Himalayan lichen Cetrelia olivetorum.

    Science.gov (United States)

    Savale, Swapnil Anil; Pol, Chaitrali Satish; Khare, Roshni; Verma, Neeraj; Gaikwad, Subhash; Mandal, Bapi; Behera, Bhaskar C

    2016-01-01

    Lichens are source of natural bioactive compounds which are traditionally used to cure a variety of ailments. The objective of this study is to assess free radical scavenging, prolyl endopeptidase inhibitory (PEPI), and antimicrobial potential of a high altitude lichen species Cetrelia olivetorum (Nyl.) W. L. Culb. & C. F. Culb (Parmeliaceae). Lichen C. olivetorum has been cultured in vitro, and optimized culture conditions were implemented in bioreactor to obtain high quantity of biomass for the study of radical scavenging, PEPI, and antimicrobial activities. Radical scavenging activity of methanol extract of Cetrelia olivetorum (MECO) was tested at 100 µg/mL, PEPI activity at 25 and 50 µg/mL, and antimicrobial activity at 5, 25, 50, and 100 µg/mL conc. All the biological activities of natural thallus extract and its derived culture extract were evaluated spectrophotometrically. Murashige and Skoog medium supplemented with 3% glucose and 100 ppb indole-3-butyric acid (IBA) supported biomass growth at flask level and yielded 5.095 g biomass in bioreactor. MECO of both the cultured and the natural lichen exhibited half inhibiting concentration (IC50) for radical scavenging activities in the range of 50-60 µg/mL, whereas the IC50 value of standard antioxidants was found to be in the range of 12-29 µg/mL. The IC50 value of lichen extract for PEPI activity was 144-288 µg/mL, whereas the IC50 value of standard prolyl endopeptidase inhibitor, Z-pro-prolinal, was 57.73 µg/mL. As far as the antimicrobial activity of MECO is concerned, minimum inhibitory concentration (MIC) value of lichen extracts against tested microorganisms was obtained in the range of 50-104 µg/mL and found to be more effective than commercially available standard erythromycin. Murashige and Skoog medium containing IBA was found to be suitable for maximum biomass production of C. olivetorum under bioreactor conditions. The cultured lichen biomass extract also showed

  5. Identification and comparative analysis of sixteen fungal peptidyl-prolyl cis/trans isomerase repertoires

    Directory of Open Access Journals (Sweden)

    Pemberton Trevor J

    2006-09-01

    Full Text Available Abstract Background The peptidyl-prolyl cis/trans isomerase (PPIase class of proteins is present in all known eukaryotes, prokaryotes, and archaea, and it is comprised of three member families that share the ability to catalyze the cis/trans isomerisation of a prolyl bond. Some fungi have been used as model systems to investigate the role of PPIases within the cell, however how representative these repertoires are of other fungi or humans has not been fully investigated. Results PPIase numbers within these fungal repertoires appears associated with genome size and orthology between repertoires was found to be low. Phylogenetic analysis showed the single-domain FKBPs to evolve prior to the multi-domain FKBPs, whereas the multi-domain cyclophilins appear to evolve throughout cyclophilin evolution. A comparison of their known functions has identified, besides a common role within protein folding, multiple roles for the cyclophilins within pre-mRNA splicing and cellular signalling, and within transcription and cell cycle regulation for the parvulins. However, no such commonality was found with the FKBPs. Twelve of the 17 human cyclophilins and both human parvulins, but only one of the 13 human FKBPs, identified orthologues within these fungi. hPar14 orthologues were restricted to the Pezizomycotina fungi, and R. oryzae is unique in the known fungi in possessing an hCyp33 orthologue and a TPR-containing FKBP. The repertoires of Cryptococcus neoformans, Aspergillus fumigatus, and Aspergillus nidulans were found to exhibit the highest orthology to the human repertoire, and Saccharomyces cerevisiae one of the lowest. Conclusion Given this data, we would hypothesize that: (i the evolution of the fungal PPIases is driven, at least in part, by the size of the proteome, (ii evolutionary pressures differ both between the different PPIase families and the different fungi, and (iii whilst the cyclophilins and parvulins have evolved to perform conserved

  6. Inhibition of prolyl 4-hydroxylase decreases muscle fibrosis following chronic rotator cuff tear.

    Science.gov (United States)

    Gumucio, J P; Flood, M D; Bedi, A; Kramer, H F; Russell, A J; Mendias, C L

    2017-01-01

    Rotator cuff tears are among the most frequent upper extremity injuries. Current treatment strategies do not address the poor quality of the muscle and tendon following chronic rotator cuff tears. Hypoxia-inducible factor-1 alpha (HIF-1α) is a transcription factor that activates many genes that are important in skeletal muscle regeneration. HIF-1α is inhibited under normal physiological conditions by the HIF prolyl 4-hydroxylases (PHDs). In this study, we used a pharmacological PHD inhibitor, GSK1120360A, to enhance the activity of HIF-1α following the repair of a chronic cuff tear, and measured muscle fibre contractility, fibrosis, gene expression, and enthesis mechanics. Chronic supraspinatus tears were induced in adult rats, and repaired 28 days later. Rats received 0 mg/kg, 3 mg/kg, or 10 mg/kg GSK1120360A daily. Collagen content, contractility, fibre type distribution and size, the expression of genes involved in fibrosis, lipid accumulation, atrophy and inflammation, and the mechanical properties of the enthesis were then assessed two weeks following surgical repair. At two weeks following repair, treatment groups showed increased muscle mass but there was a 15% decrease in force production in the 10 mg/kg group from controls, and no difference between the 0 mg/kg and the 3 mg/kg groups. There was a decrease in the expression of several gene transcripts related to matrix accumulation and fibrosis, and a 50% decrease in collagen content in both treated groups compared with controls. Additionally, the expression of inflammatory genes was reduced in the treated groups compared with controls. Finally, PHD inhibition improved the maximum stress and displacement to failure in repaired tendons. GSK1120360A resulted in improved enthesis mechanics with variable effects on muscle function. PHD inhibition may be beneficial for connective tissue injuries in which muscle atrophy has not occurred.Cite this article: J. P. Gumucio, M. D. Flood, A. Bedi, H. F. Kramer, A. J

  7. Prolyl Endopeptidase (PREP) is Associated With Male Reproductive Functions and Gamete Physiology in Mice.

    Science.gov (United States)

    Dotolo, Raffaele; Kim, Jung Dae; Pariante, Paolo; Minucci, Sergio; Diano, Sabrina

    2016-03-01

    Prolyl endopeptidase (PREP) is a serine protease which has been implicated in many biological processes, such as the maturation and degradation of peptide hormones and neuropeptides, learning and memory, cell proliferation and differentiation, and glucose metabolism. A small number of reports have also suggested PREP participation in both male and female reproduction-associated processes. In the present work, we examined PREP distribution in male germ cells and studied the effects of its knockdown (Prep(gt/gt)) on testis and sperm in adult mice. The protein is expressed and localized in elongating spermatids and luminal spermatozoa of wild type (wt) mice, as well as Sertoli, Leydig, and peritubular cells. PREP is also expressed in the head and midpiece of epididymal spermatozoa, whereas the remaining tail region shows a weaker signal. Furthermore, testis weight, histology of seminiferous tubules, and epididymal sperm parameters were assessed in wt and Prep(gt/gt) mice: wild type testes have larger average tubule and lumen diameter; in addition, lumenal composition of seminiferous tubules is dissimilar between wt and Prep(gt/gt), as the percentage of spermiated tubules is much higher in wt. Finally, total sperm count, sperm motility, and normal morphology are also higher in wt than in Prep(gt/gt). These results show for the first time that the expression of PREP could be necessary for a correct reproductive function, and suggest that the enzyme may play a role in mouse spermatogenesis and sperm physiology. © 2015 Wiley Periodicals, Inc.

  8. High Myopia Caused by a Mutation in LEPREL1, Encoding Prolyl 3-Hydroxylase 2

    Science.gov (United States)

    Mordechai, Shikma; Gradstein, Libe; Pasanen, Annika; Ofir, Rivka; El Amour, Khalil; Levy, Jaime; Belfair, Nadav; Lifshitz, Tova; Joshua, Sara; Narkis, Ginat; Elbedour, Khalil; Myllyharju, Johanna; Birk, Ohad S.

    2011-01-01

    Autosomal-recessive high-grade axial myopia was diagnosed in Bedouin Israeli consanguineous kindred. Some affected individuals also had variable expressivity of early-onset cataracts, peripheral vitreo-retinal degeneration, and secondary sight loss due to severe retinal detachments. Through genome-wide linkage analysis, the disease-associated gene was mapped to ∼1.7 Mb on chromosome 3q28 (the maximum LOD score was 11.5 at θ = 0 for marker D3S1314). Sequencing of the entire coding regions and intron-exon boundaries of the six genes within the defined locus identified a single mutation (c.1523G>T) in exon 10 of LEPREL1, encoding prolyl 3-hydroxylase 2 (P3H2), a 2-oxoglutarate-dependent dioxygenase that hydroxylates collagens. The mutation affects a glycine that is conserved within P3H isozymes. Analysis of wild-type and p.Gly508Val (c.1523G>T) mutant recombinant P3H2 polypeptides expressed in insect cells showed that the mutation led to complete inactivation of P3H2. PMID:21885030

  9. Incorporation of a prolyl hydroxylase inhibitor into scaffolds: a strategy for stimulating vascularization.

    Science.gov (United States)

    Sham, Adeline; Martinez, Eliana C; Beyer, Sebastian; Trau, Dieter W; Raghunath, Michael

    2015-03-01

    Clinical applications of tissue engineering are constrained by the ability of the implanted construct to invoke vascularization in adequate extent and velocity. To overcome the current limitations presented by local delivery of single angiogenic factors, we explored the incorporation of prolyl hydroxylase inhibitors (PHIs) into scaffolds as an alternative vascularization strategy. PHIs are small molecule drugs that can stabilize the alpha subunit of hypoxia-inducible factor-1 (HIF-1), a key transcription factor that regulates a variety of angiogenic mechanisms. In this study, we conjugated the PHI pyridine-2,4-dicarboxylic acid (PDCA) through amide bonds to a gelatin sponge (Gelfoam(®)). Fibroblasts cultured on PDCA-Gelfoam were able to infiltrate and proliferate in these scaffolds while secreting significantly more vascular endothelial growth factor than cells grown on Gelfoam without PDCA. Reporter cells expressing green fluorescent protein-tagged HIF-1α exhibited dose-dependent stabilization of this angiogenic transcription factor when growing within PDCA-Gelfoam constructs. Subsequently, we implanted PDCA-Gelfoam scaffolds into the perirenal fat tissue of Sprague Dawley rats for 8 days. Immunostaining of explants revealed that the PDCA-Gelfoam scaffolds were amply infiltrated by cells and promoted vascular ingrowth in a dose-dependent manner. Thus, the incorporation of PHIs into scaffolds appears to be a feasible strategy for improving vascularization in regenerative medicine applications.

  10. Isolation of prolyl endopeptidase inhibitory peptides from a sodium caseinate hydrolysate.

    Science.gov (United States)

    Hsieh, Cheng-Hong; Wang, Tzu-Yuan; Hung, Chuan-Chuan; Hsieh, You-Liang; Hsu, Kuo-Chiang

    2016-01-01

    Prolyl endopeptidase (PEP) has been associated with neurodegenerative disorders, and the PEP inhibitors can restore the memory loss caused by amnesic compounds. In this study, we investigated the PEP inhibitory activity of the enzymatic hydrolysates from various food protein sources, and isolated and identified the PEP inhibitory peptides. The hydrolysate obtained from sodium caseinate using bromelain (SC/BML) displayed the highest inhibitory activity of 86.8% at 5 mg mL(-1) in the present study, and its IC50 value against PEP was 0.77 mg mL(-1). The F-5 fraction by RP-HPLC (reversed-phase high performance liquid chromatography) from SC/BML showed the highest PEP inhibition rate of 88.4%, and 9 peptide sequences were identified. The synthetic peptides (1245.63-1787.94 Da) showed dose-dependent inhibition effects on PEP as competitive inhibitors with IC50 values between 29.8 and 650.5 μM. The results suggest that the peptides derived from sodium caseinate have the potential to be PEP inhibitors.

  11. Peptidyl Prolyl Isomerase PIN1 Directly Binds to and Stabilizes Hypoxia-Inducible Factor-1α.

    Directory of Open Access Journals (Sweden)

    Hyeong-Jun Han

    Full Text Available Peptidyl prolyl isomerase (PIN1 regulates the functional activity of a subset of phosphoproteins through binding to phosphorylated Ser/Thr-Pro motifs and subsequently isomerization of the phosphorylated bonds. Interestingly, PIN1 is overexpressed in many types of malignancies including breast, prostate, lung and colon cancers. However, its oncogenic functions have not been fully elucidated. Here, we report that PIN1 directly interacts with hypoxia-inducible factor (HIF-1α in human colon cancer (HCT116 cells. PIN1 binding to HIF-1α occurred in a phosphorylation-dependent manner. We also found that PIN1 interacted with HIF-1α at both exogenous and endogenous levels. Notably, PIN1 binding stabilized the HIF-1α protein, given that their levels were significantly increased under hypoxic conditions. The stabilization of HIF-1α resulted in increased transcriptional activity, consequently upregulating expression of vascular endothelial growth factor, a major contributor to angiogenesis. Silencing of PIN1 or pharmacologic inhibition of its activity abrogated the angiogenesis. By utilizing a bioluminescence imaging technique, we were able to demonstrate that PIN1 inhibition dramatically reduced the tumor volume in a subcutaneous mouse xenograft model and angiogenesis as well as hypoxia-induced transcriptional activity of HIF-1α. These results suggest that PIN1 interacting with HIF-1α is a potential cancer chemopreventive and therapeutic target.

  12. Attenuation of antagonist-induced impairment of dopamine receptors by L-prolyl-L-leucyl-glycinamide

    International Nuclear Information System (INIS)

    Saleh, M.I.M.

    1988-01-01

    The present study was undertaken in order to determine whether chronic,long-term postnatal challenge of rat pups per se, with specific dopamine D1 and D2 receptor antagonists, would modify the ontogeny of the respective receptor types. Since the neuropeptide L-prolyl-L-leucyl-glycinamide (PLG) attenuates the effect of haloperidol on dopamine D2 receptors in adult rats it was of interest to determine whether PLG would modulate antagonists-induced alterations in the ontogeny of striatal dopamine D1 and D2 receptors. Half of the rats were treated daily for 32 days from birth with SCH-23390, a selective dopamine D1 antagonist; or spiroperidol, a selective dopamine D2 antagonists; or both SCH-23390 and spiroperidol; or saline. The other half of the litters were treated with PLG, in combination with the other treatments. Animals were decapitated at 5, 8, and 12 weeks from birth for neurochemical analysis of the striatum. Chronic SCH-23390 treatment produced a 70-80% decrease in the binding of [ 3 H] SCH-23390 to striatal homogenates. The alteration at 5 weeks was associated with a 78% decrease in the Bmax for [ 3 H] SCH-23390 binding, and no change in the K D . Similarly, at 5, 8, and 12 weeks, chronic spiroperidol treatment reduced the binding of [ 3 H] spiroperidol to striatal homogenates by 70-80%

  13. Proline substitutions in a Mip-like peptidyl-prolyl cis-trans isomerase severely affect its structure, stability, shape and activity

    Directory of Open Access Journals (Sweden)

    Soumitra Polley

    2015-01-01

    Full Text Available FKBP22, an Escherichia coli-specific peptidyl-prolyl cis-trans isomerase, shows substantial homology with the Mip-like virulence factors. Mip-like proteins are homodimeric and possess a V-shaped conformation. Their N-terminal domains form dimers, whereas their C-terminal domains bind protein/peptide substrates and distinct inhibitors such as rapamycin and FK506. Interestingly, the two domains of the Mip-like proteins are separated by a lengthy, protease-susceptible α-helix. To delineate the structural requirement of this domain-connecting region in Mip-like proteins, we have investigated a recombinant FKBP22 (rFKBP22 and its three point mutants I65P, V72P and A82P using different probes. Each mutant harbors a Pro substitution mutation at a distinct location in the hinge region. We report that the three mutants are not only different from each other but also different from rFKBP22 in structure and activity. Unlike rFKBP22, the three mutants were unfolded by a non-two state mechanism in the presence of urea. In addition, the stabilities of the mutants, particularly I65P and V72P, differed considerably from that of rFKBP22. Conversely, the rapamycin binding affinity of no mutant was different from that of rFKBP22. Of the mutants, I65P showed the highest levels of structural/functional loss and dissociated partly in solution. Our computational study indicated a severe collapse of the V-shape in I65P due to the anomalous movement of its C-terminal domains. The α-helical nature of the domain-connecting region is, therefore, critical for the Mip-like proteins.

  14. A dual inhibitor against prolyl isomerase Pin1 and cyclophilin discovered by a novel real-time fluorescence detection method

    International Nuclear Information System (INIS)

    Mori, Tadashi; Hidaka, Masafumi; Lin, Yi-Chin; Yoshizawa, Ibuki; Okabe, Takayoshi; Egashira, Shinichiro; Kojima, Hirotatsu; Nagano, Tetsuo; Koketsu, Mamoru; Takamiya, Mari; Uchida, Takafumi

    2011-01-01

    Research highlights: → A Pin1 (prolyl isomerase) inhibitor, TME-001, has been discovered by using a new established high-throughput screening method. → The TME-001 showed a cell-active inhibition with lower cytotoxic effect than known Pin1 inhibitors. → Kinetic analyses revealed that the TME-001 is the first compound that exhibits dual inhibition of Pin1 and another type of prolyl isomerase, cyclophilin. → Thus, similarities of structure and reaction mechanism between Pin1 and cyclophilin are proposed. -- Abstract: Pin1, a peptidyl prolyl cis/trans isomerase (PPIase), is a potential target molecule for cancer, infectious disease, and Alzheimer's disease. We established a high-throughput screening method for Pin1 inhibitors, which employs a real-time fluorescence detector. This screening method identified 66 compounds that inhibit Pin1 out of 9756 compounds from structurally diverse chemical libraries. Further evaluations of surface plasmon resonance methods and a cell proliferation assay were performed. We discovered a cell-active inhibitor, TME-001 (2-(3-chloro-4-fluoro-phenyl)-isothiazol-3-one). Surprisingly, kinetic analyses revealed that TME-001 is the first compound that exhibits dual inhibition of Pin1 (IC 50 = 6.1 μM) and cyclophilin, another type of PPIase, (IC 50 = 13.7 μM). This compound does not inhibit FKBP. This finding suggests the existence of similarities of structure and reaction mechanism between Pin1 and cyclophilin, and may lead to a more complete understanding of the active sites of PPIases.

  15. Minoxidil Induction of VEGF Is Mediated by Inhibition of HIF-Prolyl Hydroxylase

    Directory of Open Access Journals (Sweden)

    Soohwan Yum

    2017-12-01

    Full Text Available The topical application of minoxidil may achieve millimolar concentrations in the skin. We investigated whether millimolar minoxidil could induce vascular endothelial growth factor (VEGF, a possible effector for minoxidil-mediated hair growth, and how it occurred at the molecular level. Cell-based experiments were performed to investigate a molecular mechanism underlying the millimolar minoxidil induction of VEGF. The inhibitory effect of minoxidil on hypoxia-inducible factor (HIF prolyl hydroxylase-2 (PHD-2 was tested by an in vitro von Hippel–Lindau protein (VHL binding assay. To examine the angiogenic potential of millimolar minoxidil, a chorioallantoic membrane (CAM assay was used. In human keratinocytes and dermal papilla cells, millimolar minoxidil increased the secretion of VEGF, which was not attenuated by a specific adenosine receptor antagonist that inhibits the micromolar minoxidil induction of VEGF. Millimolar minoxidil induced hypoxia-inducible factor-1α (HIF-1α, and the induction of VEGF was dependent on HIF-1. Moreover, minoxidil applied to the dorsal area of mice increased HIF-1α and VEGF in the skin. In an in vitro VHL binding assay, minoxidil directly inhibited PHD-2, thus preventing the hydroxylation of cellular HIF-1α and VHL-dependent proteasome degradation and resulting in the stabilization of HIF-1α protein. Minoxidil inhibition of PHD-2 was reversed by ascorbate, a cofactor of PHD-2, and the minoxidil induction of cellular HIF-1α was abrogated by the cofactor. Millimolar minoxidil promoted angiogenesis in the CAM assay, an in vivo angiogenic test, and this was nullified by the specific inhibition of VEGF. Our data demonstrate that PHD may be the molecular target for millimolar minoxidil-mediated VEGF induction via HIF-1.

  16. Minoxidil Induction of VEGF Is Mediated by Inhibition of HIF-Prolyl Hydroxylase

    Science.gov (United States)

    Yum, Soohwan; Jeong, Seongkeun; Kim, Dohoon; Lee, Sunyoung; Kim, Wooseong; Yoo, Jin-Wook; Kwon, Oh Sang; Kim, Dae-Duk; Min, Do Sik; Jung, Yunjin

    2017-01-01

    The topical application of minoxidil may achieve millimolar concentrations in the skin. We investigated whether millimolar minoxidil could induce vascular endothelial growth factor (VEGF), a possible effector for minoxidil-mediated hair growth, and how it occurred at the molecular level. Cell-based experiments were performed to investigate a molecular mechanism underlying the millimolar minoxidil induction of VEGF. The inhibitory effect of minoxidil on hypoxia-inducible factor (HIF) prolyl hydroxylase-2 (PHD-2) was tested by an in vitro von Hippel–Lindau protein (VHL) binding assay. To examine the angiogenic potential of millimolar minoxidil, a chorioallantoic membrane (CAM) assay was used. In human keratinocytes and dermal papilla cells, millimolar minoxidil increased the secretion of VEGF, which was not attenuated by a specific adenosine receptor antagonist that inhibits the micromolar minoxidil induction of VEGF. Millimolar minoxidil induced hypoxia-inducible factor-1α (HIF-1α), and the induction of VEGF was dependent on HIF-1. Moreover, minoxidil applied to the dorsal area of mice increased HIF-1α and VEGF in the skin. In an in vitro VHL binding assay, minoxidil directly inhibited PHD-2, thus preventing the hydroxylation of cellular HIF-1α and VHL-dependent proteasome degradation and resulting in the stabilization of HIF-1α protein. Minoxidil inhibition of PHD-2 was reversed by ascorbate, a cofactor of PHD-2, and the minoxidil induction of cellular HIF-1α was abrogated by the cofactor. Millimolar minoxidil promoted angiogenesis in the CAM assay, an in vivo angiogenic test, and this was nullified by the specific inhibition of VEGF. Our data demonstrate that PHD may be the molecular target for millimolar minoxidil-mediated VEGF induction via HIF-1. PMID:29295567

  17. Prolyl hydroxylase-1 regulates hepatocyte apoptosis in an NF-κB-dependent manner

    Energy Technology Data Exchange (ETDEWEB)

    Fitzpatrick, Susan F.; Fábián, Zsolt; Schaible, Bettina; Lenihan, Colin R.; Schwarzl, Thomas [School of Medicine and Medical Science, The Conway Institute, University College Dublin, Belfield, Dublin 4 Ireland (Ireland); Rodriguez, Javier [Systems Biology Ireland, University College Dublin, Dublin 4 (Ireland); Zheng, Xingnan; Li, Zongwei [Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC (United States); Tambuwala, Murtaza M. [School of Pharmacy and Pharmaceutical Sciences, Ulster University, Coleraine, BT52 1SA, Northern Ireland (United Kingdom); Higgins, Desmond G.; O' Meara, Yvonne [School of Medicine and Medical Science, The Conway Institute, University College Dublin, Belfield, Dublin 4 Ireland (Ireland); Slattery, Craig [School of Biomolecular and Biomedical Science, The Conway Institute, University College Dublin, Belfield, Dublin 4 Ireland (Ireland); Manresa, Mario C. [School of Medicine and Medical Science, The Conway Institute, University College Dublin, Belfield, Dublin 4 Ireland (Ireland); Fraisl, Peter; Bruning, Ulrike [Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology, University of Leuven, Vesalius Research Center, VIB, B-3000 (Belgium); Baes, Myriam [Laboratory for Cell Metabolism, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven (Belgium); Carmeliet, Peter; Doherty, Glen [Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology, University of Leuven, Vesalius Research Center, VIB, B-3000 (Belgium); Kriegsheim, Alex von [Systems Biology Ireland, University College Dublin, Dublin 4 (Ireland); Cummins, Eoin P. [School of Medicine and Medical Science, The Conway Institute, University College Dublin, Belfield, Dublin 4 Ireland (Ireland); and others

    2016-06-03

    Hepatocyte death is an important contributing factor in a number of diseases of the liver. PHD1 confers hypoxic sensitivity upon transcription factors including the hypoxia inducible factor (HIF) and nuclear factor-kappaB (NF-κB). Reduced PHD1 activity is linked to decreased apoptosis. Here, we investigated the underlying mechanism(s) in hepatocytes. Basal NF-κB activity was elevated in PHD1{sup −/−} hepatocytes compared to wild type controls. ChIP-seq analysis confirmed enhanced binding of NF-κB to chromatin in regions proximal to the promoters of genes involved in the regulation of apoptosis. Inhibition of NF-κB (but not knock-out of HIF-1 or HIF-2) reversed the anti-apoptotic effects of pharmacologic hydroxylase inhibition. We hypothesize that PHD1 inhibition leads to altered expression of NF-κB-dependent genes resulting in reduced apoptosis. This study provides new information relating to the possible mechanism of therapeutic action of hydroxylase inhibitors that has been reported in pre-clinical models of intestinal and hepatic disease. -- Highlights: •Genetic ablation of PHD1 upregulates NF-kappaB (NF-κB) in hepatocytes. •Activation of NF-κB leads to differential DNA-binding of p50/p65 and results in differential regulation of apoptotic genes. •We identified proline 191 in the beta subunit of the I-kappaB kinase as a target for PHD1-mediated hydroxylation. •Blockade of prolyl-4-hydroxylases has been found cytoprotective in liver cells.

  18. Offshore Substrate

    Data.gov (United States)

    California Natural Resource Agency — This shapefile displays the distribution of substrate types from Pt. Arena to Pt. Sal in central/northern California. Originally this data consisted of seven paper...

  19. Functional characterisation of parvulin-type peptidyl prolyl cis-trans isomerase, PinA in Dictyostelium discoideum

    International Nuclear Information System (INIS)

    Haokip, Nemneineng; Naorem, Aruna

    2017-01-01

    Pin1-type parvulins are unique among PPIases that can catalyse an otherwise slow cis-trans isomerisation of phosphorylated peptide bond preceding proline in target proteins. This prolyl isomerisation process can regulate activity, stability and localisation of target proteins and thus control cellular processes like eukaryotic cell proliferation, cell cycle progression and gene regulation. Towards understanding the function of Pin1-type prolyl isomerisation in Dictyostelium discoideum, a slime mould with distinct growth and developmental phases, we identified PinA as a novel Pin1-type parvulin by its ability to complement the temperature sensitivity phenotype associated with a mutation in ESS1 in S. cerevisiae. In D. discoideum, pinA is temporally and spatially regulated during growth and development. PinA is both nuclear as well as cytoplasmic in the growing cells. We further show that loss of pinA (pinA − ) leads to decreased growth rate, reduced spore formation and abnormal prespore-prestalk patterning. We conclude that PinA is required for normal growth as well as development in D. discoideum. - Highlights: • PinA is a bona fide homologue of S. cerevisiae Ess1. • PinA is required for normal cell proliferation of D. discoideum. • PinA is spatially localised in developmental structures. • PinA is important for cell differentiation and patterning.

  20. Human cyclophilin B: A second cyclophilin gene encodes a peptidyl-prolyl isomerase with a signal sequence

    International Nuclear Information System (INIS)

    Price, E.R.; Zydowsky, L.D.; Jin, Mingjie; Baker, C.H.; McKeon, F.D.; Walsh, C.T.

    1991-01-01

    The authors report the cloning and characterization of a cDNA encoding a second human cyclosporin A-binding protein (hCyPB). Homology analyses reveal that hCyPB is a member of the cyclophilin B (CyPB) family, which includes yeast CyPB, Drosophila nina A, and rat cyclophilin-like protein. This family is distinguished from the cyclophilin A (CyPA) family by the presence of endoplasmic reticulum (ER)-directed signal sequences. hCyPB has a hydrophobic leader sequence not found in hCyPA, and its first 25 amino acids are removed upon expression in Escherichia coli. Moreover, they show that hCyPB is a peptidyl-prolyl cis-trans isomerase which can be inhibited by cyclosporin A. These observations suggest that other members of the CyPB family will have similar enzymatic properties. Sequence comparisons of the CyPB proteins show a central, 165-amino acid peptidyl-prolyl isomerase and cyclosprorin A-binding domain, flanked by variable N-terminal and C-terminal domains. These two variable regions may impart compartmental specificity and regulation to this family of cyclophilin proteins containing the conserved core domain. Northern blot analyses show that hCyPB mRNA is expressed in the Jurkat T-cell line, consistent with its possible target role in cyclosporin A-mediated immunosuppression

  1. Manganese (II) induces chemical hypoxia by inhibiting HIF-prolyl hydroxylase: Implication in manganese-induced pulmonary inflammation

    International Nuclear Information System (INIS)

    Han, Jeongoh; Lee, Jong-Suk; Choi, Daekyu; Lee, Youna; Hong, Sungchae; Choi, Jungyun; Han, Songyi; Ko, Yujin; Kim, Jung-Ae; Mi Kim, Young; Jung, Yunjin

    2009-01-01

    Manganese (II), a transition metal, causes pulmonary inflammation upon environmental or occupational inhalation in excess. We investigated a potential molecular mechanism underlying manganese-induced pulmonary inflammation. Manganese (II) delayed HIF-1α protein disappearance, which occurred by inhibiting HIF-prolyl hydroxylase (HPH), the key enzyme for HIF-1α hydroxylation and subsequent von Hippel-Lindau(VHL)-dependent HIF-1α degradation. HPH inhibition by manganese (II) was neutralized significantly by elevated dose of iron. Consistent with this, the induction of cellular HIF-1α protein by manganese (II) was abolished by pretreatment with iron. Manganese (II) induced the HIF-1 target gene involved in pulmonary inflammation, vascular endothelial growth factor (VEGF), in lung carcinoma cell lines. The induction of VEGF was dependent on HIF-1. Manganese-induced VEGF promoted tube formation of HUVEC. Taken together, these data suggest that HIF-1 may be a potential mediator of manganese-induced pulmonary inflammation

  2. Synthesis of protected 2-pyrrolylalanine for peptide chemistry and examination of its influence on prolyl amide isomer equilibrium.

    Science.gov (United States)

    Dörr, Aurélie A; Lubell, William D

    2012-08-03

    Protected enantiopure 2-pyrrolylalanine was synthesized for application in peptide science as an electron-rich arylalanine (histidine) analog with π-donor capability. (2S)-N-(Boc)-N'-(Phenylsulfonyl)-, (2S)-N,N'-bis-(phenylsulfonyl)-, and (2S)-N,N'-bis-(Boc)-3-(2-pyrrolyl)alanines (10, 3, and 14, respectively) were made in 13-17% overall yields and six to seven steps from oxazolidine β-methyl ester 4. Homoallylic ketone 5 was prepared by a copper-catalyzed cascade addition of vinylmagnesium bromide to ester 4 and converted to pyrrolyl amino alcohol 7 by olefin oxidation and Paal-Knorr condensation. Protecting group shuffle and oxidation of the primary alcohol enabled the synthesis of pyrrolylalanines. The bis-Boc analog 14 proved useful in peptide chemistry and was employed to make N-acetyl-pyrrolylalaninyl-proline N''-methylamide 25. A study of the influence of the pyrrole moiety on the prolyl amide isomer equilibrium of 25 using (1)H NMR spectroscopy in chloroform, DMSO, and water demonstrated that the pyrrolylalanine peptide exhibited behavior and conformations different from those of other arylalanine analogs.

  3. The Cytoplasmic Prolyl-tRNA Synthetase of the Malaria Parasite is a Dual-Stage Target for Drug Development

    Science.gov (United States)

    Herman, Jonathan D.; Pepper, Lauren R.; Cortese, Joseph F.; Estiu, Guillermina; Galinsky, Kevin; Zuzarte-Luis, Vanessa; Derbyshire, Emily R.; Ribacke, Ulf; Lukens, Amanda K.; Santos, Sofia A.; Patel, Vishal; Clish, Clary B.; Sullivan, William J.; Zhou, Huihao; Bopp, Selina E.; Schimmel, Paul; Lindquist, Susan; Clardy, Jon; Mota, Maria M.; Keller, Tracy L.; Whitman, Malcolm; Wiest, Olaf; Wirth, Dyann F.; Mazitschek, Ralph

    2015-01-01

    The emergence of drug resistance is a major limitation of current antimalarials. The discovery of new druggable targets and pathways including those that are critical for multiple life cycle stages of the malaria parasite is a major goal for the development of the next-generation of antimalarial drugs. Using an integrated chemogenomics approach that combined drug-resistance selection, whole genome sequencing and an orthogonal yeast model, we demonstrate that the cytoplasmic prolyl-tRNA synthetase (PfcPRS) of the malaria parasite Plasmodium falciparum is a biochemical and functional target of febrifugine and its synthetic derivatives such as halofuginone. Febrifugine is the active principle of a traditional Chinese herbal remedy for malaria. We show that treatment with febrifugine derivatives activated the amino acid starvation response in both P. falciparum and a transgenic yeast strain expressing PfcPRS. We further demonstrate in the P. berghei mouse model of malaria that halofuginol, a new halofuginone analog that we developed, is highly active against both liver and asexual blood stages of the malaria parasite. Halofuginol, unlike halofuginone and febrifugine, is well tolerated at efficacious doses, and represents a promising lead for the development of dual-stage next generation antimalarials. PMID:25995223

  4. Oral delivery of prolyl hydroxylase inhibitor: AKB-4924 promotes localized mucosal healing in a mouse model of colitis.

    Science.gov (United States)

    Marks, Ellen; Goggins, Bridie J; Cardona, Jocelle; Cole, Siobhan; Minahan, Kyra; Mateer, Sean; Walker, Marjorie M; Shalwitz, Robert; Keely, Simon

    2015-02-01

    Pharmacological induction of hypoxia-inducible factor (HIF), a global transcriptional regulator of the hypoxic response, by prolyl hydroxylase inhibitors (PHDi) is protective in murine models of colitis, and epithelial cells are critical for the observed therapeutic efficacy. Because systemic HIF activation may lead to potentially negative off-target effects, we hypothesized that targeting epithelial HIF through oral delivery of PHDi would be sufficient to protect against colitis in a mouse model. Using a chemically induced trinitrobenzene sulfonic acid murine model of colitis, we compared the efficacy of oral and intraperitoneal (i.p.) delivery of the PHDi; AKB-4924 in preventing colitis, as measured by endoscopy, histology, barrier integrity, and immune profiling. Furthermore, we measured potential off-target effects, examining HIF and HIF target genes in the heart and kidney, as well as erythropoietin and hematocrit levels. Oral administration of AKB-4924 exhibited mucosal protection comparable i.p. dosing. Oral delivery of PHDi led to reduced colonic epithelial HIF stabilization compared with i.p. delivery, but this was still sufficient to induce transcription of downstream HIF targets. Furthermore, oral delivery of PHDi led to reduced stabilization of HIF and activation of HIF targets in extraintestinal organs. Oral delivery of PHDi therapies to this intestinal mucosa protects against colitis in animal models and represents a potential therapeutic strategy for inflammatory bowel disease, which also precludes unwanted extraintestinal effects.

  5. Characterizing the interactions between prolyl isomerase pin1 and phosphatase inhibitor-2 in living cells with FRET and FCS

    Science.gov (United States)

    Sun, Yuansheng; Wang, Lifu; Jyothikumar, Vinod; Brautigan, David L.; Periasamy, Ammasi

    2012-03-01

    Phosphatase inhibitor-2 (I2) was discovered as a regulator of protein Ser/Thr phosphatase-1 and is conserved from yeast to human. Binding between purified recombinant I2 from different species and the prolyl isomerase Pin1 has been demonstrated with pull-down assays, size exclusion chromatography and nuclear magnetic resonance spectroscopy. Despite this, questions persist as to whether these proteins associate together in living cells. In this study, we prepared fluorescent protein (FP) fusions of I2 and Pin1 and employed both Förster Resonance Energy Transfer (FRET) and Fluorescence Correlation Spectroscopy (FCS) imaging techniques to characterize their interactions in living cells. In both intensity-based and time-resolved FRET studies, we observed FRET uniformly across whole cells co-expressing I2-Cerulean and Pin1-Venus that was significantly higher than in negative controls expressing Cerulean FP (without fusing to I2) as the FRET donor and Pin1-Venus, showing a specific interaction between I2-Cerulean and Pin1-Venus in living cells. We also observed the co-diffusion of I2-Cerulean and Pin1-mCherry in Fluorescence Cross Correlation Spectroscopy (FCCS) measurements. We further showed that I2 itself as well as I2-Pin1 formed complexes in living cells (predicted from in vitro studies) via a quantitative FRET assay, and demonstrated from FCS measurements that both I2 and Pin1 (fused to Cerulean) are highly mobile in living cells.

  6. Targeting Prolyl-tRNA Synthetase to Accelerate Drug Discovery against Malaria, Leishmaniasis, Toxoplasmosis, Cryptosporidiosis, and Coccidiosis.

    Science.gov (United States)

    Jain, Vitul; Yogavel, Manickam; Kikuchi, Haruhisa; Oshima, Yoshiteru; Hariguchi, Norimitsu; Matsumoto, Makoto; Goel, Preeti; Touquet, Bastien; Jumani, Rajiv S; Tacchini-Cottier, Fabienne; Harlos, Karl; Huston, Christopher D; Hakimi, Mohamed-Ali; Sharma, Amit

    2017-10-03

    Developing anti-parasitic lead compounds that act on key vulnerabilities are necessary for new anti-infectives. Malaria, leishmaniasis, toxoplasmosis, cryptosporidiosis and coccidiosis together kill >500,000 humans annually. Their causative parasites Plasmodium, Leishmania, Toxoplasma, Cryptosporidium and Eimeria display high conservation in many housekeeping genes, suggesting that these parasites can be attacked by targeting invariant essential proteins. Here, we describe selective and potent inhibition of prolyl-tRNA synthetases (PRSs) from the above parasites using a series of quinazolinone-scaffold compounds. Our PRS-drug co-crystal structures reveal remarkable active site plasticity that accommodates diversely substituted compounds, an enzymatic feature that can be leveraged for refining drug-like properties of quinazolinones on a per parasite basis. A compound we termed In-5 exhibited a unique double conformation, enhanced drug-like properties, and cleared malaria in mice. It thus represents a new lead for optimization. Collectively, our data offer insights into the structure-guided optimization of quinazolinone-based compounds for drug development against multiple human eukaryotic pathogens. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. The Zinc Finger of Prolyl Hydroxylase Domain Protein 2 Is Essential for Efficient Hydroxylation of Hypoxia-Inducible Factor α.

    Science.gov (United States)

    Arsenault, Patrick R; Song, Daisheng; Chung, Yu Jin; Khurana, Tejvir S; Lee, Frank S

    2016-09-15

    Prolyl hydroxylase domain protein 2 (PHD2) (also known as EGLN1) is a key oxygen sensor in mammals that posttranslationally modifies hypoxia-inducible factor α (HIF-α) and targets it for degradation. In addition to its catalytic domain, PHD2 contains an evolutionarily conserved zinc finger domain, which we have previously proposed recruits PHD2 to the HSP90 pathway to promote HIF-α hydroxylation. Here, we provide evidence that this recruitment is critical both in vitro and in vivo We show that in vitro, the zinc finger can function as an autonomous recruitment domain to facilitate interaction with HIF-α. In vivo, ablation of zinc finger function by a C36S/C42S Egln1 knock-in mutation results in upregulation of the erythropoietin gene, erythrocytosis, and augmented hypoxic ventilatory response, all hallmarks of Egln1 loss of function and HIF stabilization. Hence, the zinc finger ordinarily performs a critical positive regulatory function. Intriguingly, the function of this zinc finger is impaired in high-altitude-adapted Tibetans, suggesting that their adaptation to high altitude may, in part, be due to a loss-of-function EGLN1 allele. Thus, these findings have important implications for understanding both the molecular mechanism of the hypoxic response and human adaptation to high altitude. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  8. Collagen-derived dipeptide prolyl-hydroxyproline promotes differentiation of MC3T3-E1 osteoblastic cells

    International Nuclear Information System (INIS)

    Kimira, Yoshifumi; Ogura, Kana; Taniuchi, Yuri; Kataoka, Aya; Inoue, Naoki; Sugihara, Fumihito; Nakatani, Sachie; Shimizu, Jun; Wada, Masahiro; Mano, Hiroshi

    2014-01-01

    Highlights: • Pro-Hyp did not affect MC3T3-E1 cell proliferation and matrix mineralization. • Pro-Hyp significantly increased alkaline phosphatase activity. • Pro-Hyp significantly upregulated gene expression of Runx2, Osterix, and Col1α1. - Abstract: Prolyl-hydroxyproline (Pro-Hyp) is one of the major constituents of collagen-derived dipeptides. The objective of this study was to investigate the effects of Pro-Hyp on the proliferation and differentiation of MC3T3-E1 osteoblastic cells. Addition of Pro-Hyp did not affect MC3T3-E1 cell proliferation and matrix mineralization but alkaline phosphatase activity was significantly increased. Furthermore, cells treated with Pro-Hyp significantly upregulated gene expression of Runx2, Osterix, and Col1α1. These results indicate that Pro-Hyp promotes osteoblast differentiation. This study demonstrates for the first time that Pro-Hyp has a positive effect on osteoblast differentiation with upregulation of Runx2, Osterix, and Collα1 gene expression

  9. Structure of Prolyl-tRNA Synthetase-Halofuginone Complex Provides Basis for Development of Drugs against Malaria and Toxoplasmosis.

    Science.gov (United States)

    Jain, Vitul; Yogavel, Manickam; Oshima, Yoshiteru; Kikuchi, Haruhisa; Touquet, Bastien; Hakimi, Mohamed-Ali; Sharma, Amit

    2015-05-05

    The Chinese herb Dichroa febrifuga has traditionally treated malaria-associated fever. Its active component febrifugine (FF) and derivatives such as halofuginone (HF) are potent anti-malarials. Here, we show that FF-based derivatives arrest parasite growth by direct interaction with and inhibition of the protein translation enzyme prolyl-tRNA synthetase (PRS). Dual administration of inhibitors that target different tRNA synthetases suggests high utility of these drug targets. We reveal the ternary complex structure of PRS-HF and adenosine 5'-(β,γ-imido)triphosphate where the latter facilitates HF integration into the PRS active site. Structural analyses also highlight spaces within the PRS architecture for HF derivatization of its quinazolinone, but not piperidine, moiety. We also show a remarkable ability of HF to kill the related human parasite Toxoplasma gondii, suggesting wider HF efficacy against parasitic PRSs. Hence, our cell-, enzyme-, and structure-based data on FF-based inhibitors strengthen the case for their inclusion in anti-malarial and anti-toxoplasmosis drug development efforts. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Effects of UPR and ERAD pathway on the prolyl endopeptidase production in Pichia pastoris by controlling of nitrogen source.

    Science.gov (United States)

    Wang, Xiao-Dong; Jiang, Ting; Yu, Xiao-Wei; Xu, Yan

    2017-07-01

    Prolyl endopeptidase (PEP) is very useful in various industries, while the high cost of enzyme production remains a major obstacle for its industrial applications. Pichia pastoris has been used for the PEP production; however, the fermentation process has not be investigated and little is known about the impact of excessive PEP production on the host cell physiology. Here, we optimized the nitrogen source to improve the PEP expression level and further evaluated the cellular response including UPR and ERAD. During methanol induction phase the PEP activity (1583 U/L) was increased by 1.48-fold under the optimized nitrogen concentration of NH 4 + (300 mmol/L) and casamino acids [1.0% (w/v)] in a 3-L bioreactor. Evaluated by RT-PCR the UPR and ERAD pathways were confirmed to be activated. Furthermore, a strong decrease of ERAD-related gene transcription was observed with the addition of nitrogen source, which contributed to a higher PEP expression level.

  11. Power electronics substrate for direct substrate cooling

    Science.gov (United States)

    Le, Khiet [Mission Viejo, CA; Ward, Terence G [Redondo Beach, CA; Mann, Brooks S [Redondo Beach, CA; Yankoski, Edward P [Corona, CA; Smith, Gregory S [Woodland Hills, CA

    2012-05-01

    Systems and apparatus are provided for power electronics substrates adapted for direct substrate cooling. A power electronics substrate comprises a first surface configured to have electrical circuitry disposed thereon, a second surface, and a plurality of physical features on the second surface. The physical features are configured to promote a turbulent boundary layer in a coolant impinged upon the second surface.

  12. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to isoleucyl-prolyl-proline (IPP) and valyl-prolyl-proline (VPP) and maintenance of normal blood pressure pursuant to Article 13(5) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    to isoleucyl-prolyl-proline (IPP) and valyl-prolyl-proline (VPP) and maintenance of normal blood pressure (BP). The tripeptides IPP and VPP are sufficiently characterised. Maintenance of normal blood pressure is a beneficial physiological effect. The applicant identified a total of 20 published intervention......Following an application from Valio Ltd submitted pursuant to Article 13(5) of Regulation (EC) No 1924/2006 via the Competent Authority of Finland, the Panel on Dietetic Products, Nutrition and Allergies was asked to deliver an opinion on the scientific substantiation of a health claim related...... relationship has not been established between the consumption of IPP and VPP and maintenance of normal blood pressure. © European Food Safety Authority, 2011...

  13. Mycoplasma hyopneumoniae in vitro peptidase activities: identification and cleavage of kallikrein-kinin system-like substrates.

    Science.gov (United States)

    Moitinho-Silva, Lucas; Kondo, Marcia Y; Oliveira, Lilian C G; Okamoto, Debora N; Paes, Jéssica A; Machado, Mauricio F M; Veronez, Camila L; Motta, Guacyara; Andrade, Sheila S; Juliano, Maria A; Ferreira, Henrique B; Juliano, Luiz; Gouvea, Iuri E

    2013-05-03

    Bacterial proteases are important for metabolic processes and pathogenesis in host organisms. The bacterial swine pathogen Mycoplasma hyopneumoniae has 15 putative protease-encoding genes annotated, but none of them have been functionally characterized. To identify and characterize peptidases that could be relevant for infection of swine hosts, we investigated the peptidase activity present in the pathogenic 7448 strain of M. hyopneumoniae. Combinatorial libraries of fluorescence resonance energy transfer peptides, specific inhibitors and pH profiling were used to screen and characterize endopeptidase, aminopeptidase and carboxypeptidase activities in cell lysates. One metalloendopeptidase, one serine endopeptidase, and one aminopeptidase were detected. The detected metalloendopeptidase activity, prominent at neutral and basic pH ranges, was due to a thimet oligopeptidase family member (M3 family), likely an oligoendopeptidase F (PepF), which cleaved the peptide Abz-GFSPFRQ-EDDnp at the F-S bond. A chymotrypsin-like serine endopeptidase activity, possibly a subtilisin-like serine protease, was prominent at higher pH levels, and was characterized by its preference for a Phe residue at the P1 position of the substrate. The aminopeptidase P (APP) activity showed a similar profile to that of human membrane-bound APP. Genes coding for these three peptidases were identified and their transcription was confirmed in the 7448 strain. Furthermore, M. hyopneumoniae cell lysate peptidases showed effects on kallikrein-kinin system-like substrates, such as bradykinin-derived substrates and human high molecular weight kininogen. The M. hyopneumoniae peptidase activities, here characterized for the first time, may be important for bacterial survival strategies and thus represent possible targets for drug development against M. hyopneumoniae swine infections. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Molecular Cloning, Characterization, and Expression Analysis of a Prolyl 4-Hydroxylase from the Marine Sponge Chondrosia reniformis.

    Science.gov (United States)

    Pozzolini, Marina; Scarfì, Sonia; Mussino, Francesca; Ferrando, Sara; Gallus, Lorenzo; Giovine, Marco

    2015-08-01

    Prolyl 4-hydroxylase (P4H) catalyzes the hydroxylation of proline residues in collagen. P4H has two functional subunits, α and β. Here, we report the cDNA cloning, characterization, and expression analysis of the α and β subunits of the P4H derived from the marine sponge Chondrosia reniformis. The amino acid sequence of the α subunit is 533 residues long with an M r of 59.14 kDa, while the β subunit counts 526 residues with an M r of 58.75 kDa. Phylogenetic analyses showed that αP4H and βP4H are more related to the mammalian sequences than to known invertebrate P4Hs. Western blot analysis of sponge lysate protein cross-linking revealed a band of 240 kDa corresponding to an α2β2 tetramer structure. This result suggests that P4H from marine sponges shares the same quaternary structure with vertebrate homologous enzymes. Gene expression analyses showed that αP4H transcript is higher in the choanosome than in the ectosome, while the study of factors affecting its expression in sponge fragmorphs revealed that soluble silicates had no effect on the αP4H levels, whereas ascorbic acid strongly upregulated the αP4H mRNA. Finally, treatment with two different tumor necrosis factor (TNF)-alpha inhibitors determined a significant downregulation of αP4H gene expression in fragmorphs demonstrating, for the first time in Porifera, a positive involvement of TNF in sponge matrix biosynthesis. The molecular characterization of P4H genes involved in collagen hydroxylation, including the mechanisms that regulate their expression, is a key step for future recombinant sponge collagen production and may be pivotal to understand pathological mechanisms related to extracellular matrix deposition in higher organisms.

  15. Insights on the evolution of prolyl 3-hydroxylation sites from comparative analysis of chicken and Xenopus fibrillar collagens.

    Science.gov (United States)

    Hudson, David M; Weis, Maryann; Eyre, David R

    2011-05-03

    Recessive mutations that prevent 3-hydroxyproline formation in type I collagen have been shown to cause forms of osteogenesis imperfecta. In mammals, all A-clade collagen chains with a GPP sequence at the A1 site (P986), except α1(III), have 3Hyp at residue P986. Available avian, amphibian and reptilian type III collagen sequences from the genomic database (Ensembl) all differ in sequence motif from mammals at the A1 site. This suggests a potential evolutionary distinction in prolyl 3-hydroxylation between mammals and earlier vertebrates. Using peptide mass spectrometry, we confirmed that this 3Hyp site is fully occupied in α1(III) from an amphibian, Xenopus laevis, as it is in chicken. A thorough characterization of all predicted 3Hyp sites in collagen types I, II, III and V from chicken and xenopus revealed further differences in the pattern of occupancy of the A3 site (P707). In mammals only α2(I) and α2(V) chains had any 3Hyp at the A3 site, whereas in chicken all α-chains except α1(III) had A3 at least partially 3-hydroxylated. The A3 site was also partially 3-hydroxylated in xenopus α1(I). Minor differences in covalent cross-linking between chicken, xenopus and mammal type I and III collagens were also found as a potential index of evolving functional differences. The function of 3Hyp is still unknown but observed differences in site occupancy during vertebrate evolution are likely to give important clues.

  16. Effects of Prolyl Hydroxylase Inhibitor L-mimosine on Dental Pulp in the Presence of Advanced Glycation End Products.

    Science.gov (United States)

    Müller, Heinz-Dieter; Cvikl, Barbara; Janjić, Klara; Nürnberger, Sylvia; Moritz, Andreas; Gruber, Reinhard; Agis, Hermann

    2015-11-01

    Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  17. Peptidyl-prolyl cis/trans-isomerase A1 (Pin1) is a target for modification by lipid electrophiles.

    Science.gov (United States)

    Aluise, Christopher D; Rose, Kristie; Boiani, Mariana; Reyzer, Michelle L; Manna, Joseph D; Tallman, Keri; Porter, Ned A; Marnett, Lawrence J

    2013-02-18

    Oxidation of membrane phospholipids is associated with inflammation, neurodegenerative disease, and cancer. Oxyradical damage to phospholipids results in the production of reactive aldehydes that adduct proteins and modulate their function. 4-Hydroxynonenal (HNE), a common product of oxidative damage to lipids, adducts proteins at exposed Cys, His, or Lys residues. Here, we demonstrate that peptidyl-prolyl cis/trans-isomerase A1 (Pin1), an enzyme that catalyzes the conversion of the peptide bond of pSer/pThr-Pro moieties in signaling proteins from cis to trans, is highly susceptible to HNE modification. Incubation of purified Pin1 with HNE followed by MALDI-TOF/TOF mass spectrometry resulted in detection of Michael adducts at the active site residues His-157 and Cys-113. Time and concentration dependencies indicate that Cys-113 is the primary site of HNE modification. Pin1 was adducted in MDA-MB-231 breast cancer cells treated with 8-alkynyl-HNE as judged by click chemistry conjugation with biotin followed by streptavidin-based pulldown and Western blotting with anti-Pin1 antibody. Furthermore, orbitrap MS data support the adduction of Cys-113 in the Pin1 active site upon HNE treatment of MDA-MB-231 cells. siRNA knockdown of Pin1 in MDA-MB-231 cells partially protected the cells from HNE-induced toxicity. Recent studies indicate that Pin1 is an important molecular target for the chemopreventive effects of green tea polyphenols. The present study establishes that it is also a target for electrophilic modification by products of lipid peroxidation.

  18. Insights on the evolution of prolyl 3-hydroxylation sites from comparative analysis of chicken and Xenopus fibrillar collagens.

    Directory of Open Access Journals (Sweden)

    David M Hudson

    2011-05-01

    Full Text Available Recessive mutations that prevent 3-hydroxyproline formation in type I collagen have been shown to cause forms of osteogenesis imperfecta. In mammals, all A-clade collagen chains with a GPP sequence at the A1 site (P986, except α1(III, have 3Hyp at residue P986. Available avian, amphibian and reptilian type III collagen sequences from the genomic database (Ensembl all differ in sequence motif from mammals at the A1 site. This suggests a potential evolutionary distinction in prolyl 3-hydroxylation between mammals and earlier vertebrates. Using peptide mass spectrometry, we confirmed that this 3Hyp site is fully occupied in α1(III from an amphibian, Xenopus laevis, as it is in chicken. A thorough characterization of all predicted 3Hyp sites in collagen types I, II, III and V from chicken and xenopus revealed further differences in the pattern of occupancy of the A3 site (P707. In mammals only α2(I and α2(V chains had any 3Hyp at the A3 site, whereas in chicken all α-chains except α1(III had A3 at least partially 3-hydroxylated. The A3 site was also partially 3-hydroxylated in xenopus α1(I. Minor differences in covalent cross-linking between chicken, xenopus and mammal type I and III collagens were also found as a potential index of evolving functional differences. The function of 3Hyp is still unknown but observed differences in site occupancy during vertebrate evolution are likely to give important clues.

  19. Essential roles of insulin, AMPK signaling and lysyl and prolyl hydroxylases in the biosynthesis and multimerization of adiponectin.

    Science.gov (United States)

    Zhang, Lin; Li, Ming-Ming; Corcoran, Marie; Zhang, Shaoping; Cooper, Garth J S

    2015-01-05

    Post-translational modifications (PTMs) of the adiponectin molecule are essential for its full bioactivity, and defects in PTMs leading to its defective production and multimerization have been linked to the mechanisms of insulin resistance, obesity, and type-2 diabetes. Here we observed that, in differentiated 3T3-L1 adipocytes, decreased insulin signaling caused by blocking of insulin receptors (InsR) with an anti-InsR blocking antibody, increased rates of adiponectin secretion, whereas concomitant elevations in insulin levels counteracted this effect. Adenosine monophosphate-activated protein kinase (AMPK) signaling regulated adiponectin production by modulating the expression of adiponectin receptors, the secretion of adiponectin, and eventually the expression of adiponectin itself. We found that lysyl hydroxylases (LHs) and prolyl hydroxylases (PHs) were expressed in white-adipose tissue of ob/ob mice, wherein LH3 levels were increased compared with controls. In differentiated 3T3-L1 adipocytes, both non-specific inhibition of LHs and PHs by dipyridyl, and specific inhibition of LHs by minoxidil and of P4H with ethyl-3,4-dihydroxybenzoate, caused significant suppression of adiponectin production, more particularly of the higher-order isoforms. Transient gene knock-down of LH3 (Plod3) caused a suppressive effect, especially on the high molecular-weight (HMW) isoforms. These data indicate that PHs and LHs are both required for physiological adiponectin production and in particular are essential for the formation/secretion of the HMW isoforms. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  20. Statistical optimization of cell disruption techniques for releasing intracellular X-prolyl dipeptidyl aminopeptidase from Lactococcus lactis spp. lactis.

    Science.gov (United States)

    Üstün-Aytekin, Özlem; Arısoy, Sevda; Aytekin, Ali Özhan; Yıldız, Ece

    2016-03-01

    X-prolyl dipeptidyl aminopeptidase (PepX) is an intracellular enzyme from the Gram-positive bacterium Lactococcus lactis spp. lactis NRRL B-1821, and it has commercial importance. The objective of this study was to compare the effects of several cell disruption methods on the activity of PepX. Statistical optimization methods were performed for two cavitation methods, hydrodynamic (high-pressure homogenization) and acoustic (sonication), to determine the more appropriate disruption method. Two level factorial design (2FI), with the parameters of number of cycles and pressure, and Box-Behnken design (BBD), with the parameters of cycle, sonication time, and power, were used for the optimization of the high-pressure homogenization and sonication methods, respectively. In addition, disruption methods, consisting of lysozyme, bead milling, heat treatment, freeze-thawing, liquid nitrogen, ethylenediaminetetraacetic acid (EDTA), Triton-X, sodium dodecyl sulfate (SDS), chloroform, and antibiotics, were performed and compared with the high-pressure homogenization and sonication methods. The optimized values of high-pressure homogenization were one cycle at 130 MPa providing activity of 114.47 mU ml(-1), while sonication afforded an activity of 145.09 mU ml(-1) at 28 min with 91% power and three cycles. In conclusion, sonication was the more effective disruption method, and its optimal operation parameters were manifested for the release of intracellular enzyme from a L. lactis spp. lactis strain, which is a Gram-positive bacterium. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Ser46 phosphorylation and prolyl-isomerase Pin1-mediated isomerization of p53 are key events in p53-dependent apoptosis induced by mutant huntingtin.

    Science.gov (United States)

    Grison, Alice; Mantovani, Fiamma; Comel, Anna; Agostoni, Elena; Gustincich, Stefano; Persichetti, Francesca; Del Sal, Giannino

    2011-11-01

    Huntington disease (HD) is a neurodegenerative disorder caused by a CAG repeat expansion in the gene coding for huntingtin protein. Several mechanisms have been proposed by which mutant huntingtin (mHtt) may trigger striatal neurodegeneration, including mitochondrial dysfunction, oxidative stress, and apoptosis. Furthermore, mHtt induces DNA damage and activates a stress response. In this context, p53 plays a crucial role in mediating mHtt toxic effects. Here we have dissected the pathway of p53 activation by mHtt in human neuronal cells and in HD mice, with the aim of highlighting critical nodes that may be pharmacologically manipulated for therapeutic intervention. We demonstrate that expression of mHtt causes increased phosphorylation of p53 on Ser46, leading to its interaction with phosphorylation-dependent prolyl isomerase Pin1 and consequent dissociation from the apoptosis inhibitor iASPP, thereby inducing the expression of apoptotic target genes. Inhibition of Ser46 phosphorylation by targeting homeodomain-interacting protein kinase 2 (HIPK2), PKCδ, or ataxia telangiectasia mutated kinase, as well as inhibition of the prolyl isomerase Pin1, prevents mHtt-dependent apoptosis of neuronal cells. These results provide a rationale for the use of small-molecule inhibitors of stress-responsive protein kinases and Pin1 as a potential therapeutic strategy for HD treatment.

  2. Critical Determinants of Substrate Recognition by Cyclin-Dependent Kinase-like 5 (CDKL5).

    Science.gov (United States)

    Katayama, Syouichi; Sueyoshi, Noriyuki; Kameshita, Isamu

    2015-05-19

    Cyclin-dependent kinase-like 5 (CDKL5) is a Ser/Thr protein kinase known to be associated with X-linked neurodevelopmental disorders. In a previous study, we identified amphiphysin 1 (Amph1) as a potential substrate for CDKL5 and identified a single phosphorylation site at Ser-293. In this study, we investigated the molecular mechanisms of substrate recognition by CDKL5 using Amph1 as a model substrate. Amph1 served as an efficient CDKL5 substrate, whereas Amph2, a structurally related homologue of Amph1, was not phosphorylated by CDKL5. The sequence around the Amph1 phosphorylation site is RPR(293)SPSQ, while the corresponding sequence in Amph2 is IPK(332)SPSQ. To define the amino acid sequence specificity of the substrate, various point mutants of Amph1 and Amph2 were prepared and phosphorylated by CDKL5. Both Amph2(I329R) and Amph1 served as efficient CDKL5 substrates, but Amph1(R290I) did not, indicating that the arginyl residue at the P -3 position is critical for substrate recognition. With regard to prolyl residues around the phosphorylation site of Amph1, Pro-291 at the P -2 position, but not Pro-294 at the P +1 position, is indispensable for phosphorylation by CDKL5. Phosphorylation experiments using various deletion mutants of Amph1 revealed that the proline-rich domain (PRD) (amino acids 247-315) alone was not phosphorylated by CDKL5. In contrast, Amph1(247-385), which comprised the PRD and CLAP domains, served as an efficient CDKL5 substrate. These results, taken together, suggest that both the phosphorylation site sequence (RPXSX) and the CLAP domain structure in Amph1 play crucial roles in recognition and phosphorylation by CDKL5.

  3. Three extracellular dipeptidyl peptidases found in Aspergillus oryzae show varying substrate specificities.

    Science.gov (United States)

    Maeda, Hiroshi; Sakai, Daisuke; Kobayashi, Takuji; Morita, Hiroto; Okamoto, Ayako; Takeuchi, Michio; Kusumoto, Ken-Ichi; Amano, Hitoshi; Ishida, Hiroki; Yamagata, Youhei

    2016-06-01

    Three extracellular dipeptidyl peptidase genes, dppB, dppE, and dppF, were unveiled by sequence analysis of the Aspergillus oryzae genome. We investigated their differential enzymatic profiles, in order to gain an understanding of the diversity of these genes. The three dipeptidyl peptidases were expressed using Aspergillus nidulans as the host. Each recombinant enzyme was purified and subsequently characterized. The enzymes displayed similar optimum pH values, but optimum temperatures, pH stabilities, and substrate specificities varied. DppB was identified as a Xaa-Prolyl dipeptidyl peptidase, while DppE scissile substrates were similar to the substrates for Aspergillus fumigatus DPPV (AfDPPV). DppF was found to be a novel enzyme that could digest both substrates for A. fumigatus DPPIV and AfDPPV. Semi-quantitative PCR revealed that the transcription of dppB in A. oryzae was induced by protein substrates and repressed by the addition of an inorganic nitrogen source, despite the presence of protein substrates. The transcription of dppE depended on its growth time, while the transcription of dppF was not affected by the type of the nitrogen source in the medium, and it started during the early stage of the fungal growth. Based on these results, we conclude that these enzymes may represent the nutrition acquisition enzymes. Additionally, DppF may be one of the sensor peptidases responsible for the detection of the protein substrates in A. oryzae environment. DppB may be involved in nitrogen assimilation control, since the transcription of dppB was repressed by NaNO3, despite the presence of protein substrates.

  4. A high-throughput screen for inhibitors of the prolyl isomerase, Pin1, identifies a seaweed polyphenol that reduces adipose cell differentiation.

    Science.gov (United States)

    Mori, Tadashi; Hidaka, Masafumi; Ikuji, Hiroko; Yoshizawa, Ibuki; Toyohara, Haruhiko; Okuda, Toru; Uchida, Chiyoko; Asano, Tomoichiro; Yotsu-Yamashita, Mari; Uchida, Takafumi

    2014-01-01

    The peptidyl prolyl cis/trans isomerase Pin1 enhances the uptake of triglycerides and the differentiation of fibroblasts into adipose cells in response to insulin stimulation. Pin1 downregulation could be a potential approach to prevent and treat obesity-related disorders. In order to identify an inhibitor of Pin1 that exhibited minimal cytotoxicity, we established a high-throughput screen for Pin1 inhibitors and used this method to identify an inhibitor from 1,056 crude fractions of two natural product libraries. The candidate, a phlorotannin called 974-B, was isolated from the seaweed, Ecklonia kurome. 974-B inhibited the differentiation of mouse embryonic fibroblasts and 3T3-L1 cells into adipose cells without inducing cytotoxicity. We discovered the Pin1 inhibitor, 974-B, from the seaweed, E. kurome, and showed that it blocks the differentiation of fibroblasts into adipose cells, suggesting that 974-B could be a lead drug candidate for obesity-related disorders.

  5. Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics.

    Science.gov (United States)

    Sebela, Marek; Rehulka, Pavel; Kábrt, Jaromír; Rehulková, Helena; Ozdian, Tomás; Raus, Martin; Franc, Vojtech; Chmelík, Josef

    2009-11-01

    An acidic prolyl endoprotease from Aspergillus niger was isolated from the commercial product Brewers Clarex to evaluate its possible application in proteomics. The chromatographic purification yielded a single protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis providing an apparent molecular mass of 63 kDa and a broad peak (m/z 58,061) in linear matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) indicating the glycoprotein nature of the enzyme. Indeed, a colorimetric assessment with phenol and sulfuric acid showed the presence of neutral sugars (9% of weight). The subsequent treatment with N-glycosidase F released a variety of high-mannose type N-glycans, which were successfully detected using MALDI-TOF MS. MALDI-TOF/TOF tandem MS analysis of glycopeptides from a tryptic digest of prolyl endoprotease unraveled the identity of the N-glycosylation site in the primary structure. The data obtained also show that the enzyme is present in its processed form, i.e. without putative signal and propeptide parts. Spectrophotometric measurements demonstrated optimal activity at pH 4.0-4.5 and also high thermostability for the cleavage at the C-terminal part of proline residues. In-solution digestion of standard proteins (12-200 kDa) allowed to evaluate the cleavage specificity. The enzyme acts upon proline and alanine residues, but there is an additional minor cleavage at some other residues like Gly, Leu, Arg, Ser and Tyr. The digestion of a honeybee peptide comprising six proline residues (apidaecin 1A) led to the detection of specific peptides terminated by proline as it was confirmed by MALDI postsource decay analysis. Copyright 2009 John Wiley & Sons, Ltd.

  6. Sensor Substrate Development

    Data.gov (United States)

    National Aeronautics and Space Administration — Novel substrates, such as aerogels and porous, low density ceramics may increase the sensitivities of chemical reaction-based sensors for toxic vapors. These sensors...

  7. Isolation and characterization of cyclo-(tryptophanyl-prolyl and chloramphenicol from Streptomyces sp. SUK 25 with antimethicillin-resistant Staphylococcus aureus activity

    Directory of Open Access Journals (Sweden)

    Alshaibani MM

    2016-05-01

    resonance. Results: During screening procedure, SUK 25 exhibited good antimicrobial potential against several strains of MRSA. The best biological activity was shown from fraction number VII and its subfractions F2 and F3 with minimum inhibitory concentration values at 16 µg/mL and 8 µg/mL, respectively. These two subfractions were identified as diketopiperazine cyclo-(tryptophanyl-prolyl and chloramphenicol. Conclusion: On the basis of obtained results, SUK 25 isolated from Z. spectabile can be regarded as a new valuable source to produce secondary metabolites against bacteria, especially MRSA. Keywords: Zingiber spectabile, MRSA, SUK 25, cyclo-(tryptophanyl-prolyl, chloramphenicol, HR-MS, NMR

  8. Prolyl isomerase Pin1 is highly expressed in Her2-positive breast cancer and regulates erbB2 protein stability

    Directory of Open Access Journals (Sweden)

    Lu Kun

    2008-12-01

    Full Text Available Abstract Overexpression of HER-2/Neu occurs in about 25–30% of breast cancer patients and is indicative of poor prognosis. While Her2/Neu overexpression is primarily a result of erbB2 amplification, it has recently been recognized that erbB2 levels are also regulated on the protein level. However, factors that regulate Her2/Neu protein stability are less well understood. The prolyl isomerase Pin1 catalyzes the isomerization of specific pSer/Thr-Pro motifs that have been phosphorylated in response to mitogenic signaling. We have previously reported that Pin1-catalyzed post-phosphorylational modification of signal transduction modulates the oncogenic pathways downstream from c-neu. The goal of this study was to examine the expression of prolyl isomerase Pin1 in human Her2+ breast cancer, and to study if Pin1 affects the expression of Her2/Neu itself. Methods Immunohistochemistry for Her2 and Pin1 were performed on two hundred twenty-three human breast cancers, with 59% of the specimen from primary cancers and 41% from metastatic sites. Pin1 inhibition was achieved using siRNA in Her2+ breast cancer cell lines, and its effects were studied using cell viability assays, immunoblotting and immunofluorescence. Results Sixty-four samples (28.7% stained positive for Her2 (IHC 3+, and 54% (122/223 of all breast cancers stained positive for Pin1. Of the Her2-positive cancers 40 (62.5% were also Pin1-positive, based on strong nuclear or nuclear and cytoplasmic staining. Inhibition of Pin1 via RNAi resulted in significant suppression of Her2-positive tumor cell growth in BT474, SKBR3 and AU565 cells. Pin1 inhibition greatly increased the sensitivity of Her2-positive breast cancer cells to the mTOR inhibitor Rapamycin, while it did not increase their sensitivity to Trastuzumab, suggesting that Pin1 might act on Her2 signaling. We found that Pin1 interacted with the protein complex that contains ubiquitinated erbB2 and that Pin1 inhibition accelerated erbB2

  9. Phospho-carboxyl-terminal domain binding and the role of a prolyl isomerase in pre-mRNA 3'-End formation.

    Science.gov (United States)

    Morris, D P; Phatnani, H P; Greenleaf, A L

    1999-10-29

    A phospho-carboxyl-terminal domain (CTD) affinity column created with yeast CTD kinase I and the CTD of RNA polymerase II was used to identify Ess1/Pin1 as a phospho-CTD-binding protein. Ess1/Pin1 is a peptidyl prolyl isomerase involved in both mitotic regulation and pre-mRNA 3'-end formation. Like native Ess1, a GSTEss1 fusion protein associates specifically with the phosphorylated but not with the unphosphorylated CTD. Further, hyperphosphorylated RNA polymerase II appears to be the dominant Ess1 binding protein in total yeast extracts. We demonstrate that phospho-CTD binding is mediated by the small WW domain of Ess1 rather than the isomerase domain. These findings suggest a mechanism in which the WW domain binds the phosphorylated CTD of elongating RNA polymerase II and the isomerase domain reconfigures the CTD though isomerization of proline residues perhaps by a processive mechanism. This process may be linked to a variety of pre-mRNA maturation events that use the phosphorylated CTD, including the coupled processes of pre-mRNA 3'-end formation and transcription termination.

  10. The peptidyl prolyl cis/trans isomerase Pin1/Ess1 inhibits phosphorylation and toxicity of tau in a yeast model for Alzheimer’s disease

    Directory of Open Access Journals (Sweden)

    Ann De Vos

    2015-04-01

    Full Text Available Since hyperphosphorylation of protein tau is a crucial event in Alzheimer’s disease, additional mechanisms besides the interplay of kinase and phosphatase activities are investigated, such as the effect of the peptidyl prolyl cis/trans isomerase Pin1. This isomerase was shown to bind and isomerize phosphorylated protein tau, thereby restoring the microtubule associated protein function of tau as well as promoting the dephosphorylation of the protein by the trans-dependent phosphatase PP2A. In this study we used models based on Saccharomyces cerevisiae to further elucidate the influence of Pin1 and its yeast ortholog Ess1 on tau phosphorylation and self-assembly. We could demonstrate that in yeast, a lack of Pin1 isomerase activity leads to an increase in phosphorylation of tau at Thr231, comparable to AD brain and consistent with earlier findings in other model organisms. However, we could also distinguish an effect by Pin1 on other residues of tau, i.e. Ser235 and Ser198/199/202. Furthermore, depletion of Pin1 isomerase activity results in reduced growth of the yeast cells, which is enhanced upon expression of tau. This suggests that the accumulation of hyperphosphorylated and aggregation-prone tau causes cytotoxicity in yeast. This study introduces yeast as a valuable model organism to characterize in detail the effect of Pin1 on the biochemical characteristics of protein tau, more specifically its phosphorylation and aggregation.

  11. Changes in Hypoxia-Inducible Factor-1 (HIF-1) and Regulatory Prolyl Hydroxylase (PHD) Enzymes Following Hypoxic-Ischemic Injury in the Neonatal Rat.

    Science.gov (United States)

    Chu, Hannah X; Jones, Nicole M

    2016-03-01

    Hypoxia leads to activation of many cellular adaptive processes which are regulated by the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 consists of HIF-1α and HIF-1ß subunits and levels of HIF-1α protein are regulated by HIF prolyl-hydroxylase enzymes (PHD1, 2, 3). The aim of the current study was to investigate the expression of HIF-1α and PHDs at various time points after hypoxia-ischemia (HI), using a neonatal rat model of HI brain injury. Sprague-Dawley rat pups (postnatal day 7) were anaesthetized and underwent right carotid artery occlusion and were then exposed to 6 % oxygen for 2.5 h at 37 °C. HI injured animals demonstrated a significant reduction in the size of the ipsilateral hemisphere, compared to sham controls. Protein analysis using western blotting and enzyme-linked immunosorbent assay showed that 24 h after HI, there was a significant increase in PHD3 protein and an increase of HIF-1α compared to controls. At the 72 h time point, there was a reduction in PHD3 protein, which appeared to relate to cellular loss. There were no changes in PHD1 or PHD2 protein levels after HI when compared to age-matched controls. Further studies are necessary to establish roles for the HIF-1 regulatory enzyme PHD3 in brain injury processes.

  12. β-N-oxalyl-L-α, β- diaminopropionic acid induces HRE expression by inhibiting HIF-prolyl hydroxylase-2 in normoxic conditions.

    Science.gov (United States)

    Eslavath, Ravi Kumar; Sharma, Deepshikha; Bin Omar, Nabil A M; Chikati, Rajasekhar; Teli, Mahesh Kumar; Rajanikant, G K; Singh, Surya S

    2016-11-15

    Hypoxia inducible factor (HIF)-1α, a subunit of HIF transcription factor, regulates cellular response to hypoxia. In normoxic conditions, it is hydroxylated by prolyl hydroxylase (PHD)-2 and targeted for proteosomal degradation. Drugs which inhibit PHD-2 have implications in conditions arising from insufficient blood supply. β-ODAP (β-N- oxalyl-L-α, β- diaminopropionic acid), a non-protein excitatory amino acid present in Lathyrus sativus, is an α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor agonist known to activate conventional protein kinase C and stabilize HIF-1α under normoxic conditions. However, the mechanism of HIF-1α stabilization by this compound is unknown. In silico approach was used to understand the mechanism of stabilization of HIF-1α which revealed β-ODAP interacts with key amino acid residues and Fe 2+ at the catalytic site of PHD-2. These results were further corroborated with luciferase HRE (hypoxia response element) reporter system in HeLa cells. Different chemical modulators of PHD-2 activity and HIF-1α levels were included in the study for comparison. Results obtained indicate that β-ODAP inhibits PHD-2 and facilitates HIF dependent HRE expression and hence, might be helpful in conditions arising from hypoxia. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Coating of substrates

    International Nuclear Information System (INIS)

    Cairns, J.A.; Nelson, R.L.; Woodhead, J.L.

    1979-01-01

    The process is concerned with providing substrates with coatings obtainable from sols, for example to protect the substrate (such as in nuclear reactors or hydrocarbon cracking plant) or to provide a carrier for catalytically active material. Hitherto, coatings obtained from sols have had a high porosity and high surface area so that they have not been entirely satisfactory for the above applications. In the process described, dense, low-porosity coatings are provided by contacting the substrate with a sol of refractory material (e.g. CeO 2 or SiO 2 ) convertible to a gel of density at least 40% of the theoretical density of the refractory material, and converting the sol to the gel. Optionally, the gel may be converted to a ceramic coating by firing. (author)

  14. Robust plasmonic substrates

    DEFF Research Database (Denmark)

    Kostiučenko, Oksana; Fiutowski, Jacek; Tamulevicius, Tomas

    2014-01-01

    Robustness is a key issue for the applications of plasmonic substrates such as tip-enhanced Raman spectroscopy, surface-enhanced spectroscopies, enhanced optical biosensing, optical and optoelectronic plasmonic nanosensors and others. A novel approach for the fabrication of robust plasmonic...... substrates is presented, which relies on the coverage of gold nanostructures with diamond-like carbon (DLC) thin films of thicknesses 25, 55 and 105 nm. DLC thin films were grown by direct hydrocarbon ion beam deposition. In order to find the optimum balance between optical and mechanical properties...

  15. Substrate system for spray forming

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Men G. (Export, PA); Chernicoff, William P. (Harrisburg, PA)

    2002-01-01

    A substrate system for receiving a deposit of sprayed metal droplets including a movable outer substrate on which the sprayed metal droplets are deposited. The substrate system also includes an inner substrate disposed adjacent the outer substrate where the sprayed metal droplets are deposited on the outer substrate. The inner substrate includes zones of differing thermal conductivity to resist substrate layer porosity and to resist formation of large grains and coarse constituent particles in a bulk layer of the metal droplets which have accumulated on the outer substrate. A spray forming apparatus and associated method of spray forming a molten metal to form a metal product using the substrate system of the invention is also provided.

  16. Wetting on structured substrates

    International Nuclear Information System (INIS)

    Dietrich, S; Popescu, M N; Rauscher, M

    2005-01-01

    Chemically patterned surfaces are of significant interest in the context of microfluidic applications, and miniaturization of such devices aims at generating structures on the nano-scale. Whereas on the micron scale purely macroscopic descriptions of liquid flow are valid, on the nanometre scale long-ranged inter-molecular interactions, thermal fluctuations such as capillary waves, and finally the molecular structure of the liquid become important. We discuss the most important conceptual differences between flow on chemically patterned substrates on the micron scale and on the nanometre scale, and formulate four design issues for nanofluidics related to channel width, channel separation, and channel bending radius. As a specific example of nano-scale transport we present a microscopic model for the dynamics of spreading of monolayers on homogeneous substrates. Kinetic Monte Carlo simulations of this model on a homogeneous substrate reveal a complex spatio-temporal structure of the extracted monolayer, which includes the emergence of interfaces and of scaling properties of density profiles. These features are discussed and rationalized within the corresponding continuum limit derived from the microscopic dynamics. The corresponding spreading behaviour on a patterned substrate is briefly addressed

  17. TAL effectors target the C-terminal domain of RNA polymerase II (CTD by inhibiting the prolyl-isomerase activity of a CTD-associated cyclophilin.

    Directory of Open Access Journals (Sweden)

    Mariane Noronha Domingues

    Full Text Available Transcriptional activator-like (TAL effectors of plant pathogenic bacteria function as transcription factors in plant cells. However, how TAL effectors control transcription in the host is presently unknown. Previously, we showed that TAL effectors of the citrus canker pathogen Xanthomonas citri, named PthAs, targeted the citrus protein complex comprising the thioredoxin CsTdx, ubiquitin-conjugating enzymes CsUev/Ubc13 and cyclophilin CsCyp. Here we show that CsCyp complements the function of Cpr1 and Ess1, two yeast cyclophilins that regulate transcription by the isomerization of proline residues of the regulatory C-terminal domain (CTD of RNA polymerase II. We also demonstrate that CsCyp, CsTdx, CsUev and four PthA variants interact with the citrus CTD and that CsCyp co-immunoprecipitate with the CTD in citrus cell extracts and with PthA2 transiently expressed in sweet orange epicotyls. The interactions of CsCyp with the CTD and PthA2 were inhibited by cyclosporin A (CsA, a cyclophilin inhibitor. Moreover, we present evidence that PthA2 inhibits the peptidyl-prolyl cis-trans isomerase (PPIase activity of CsCyp in a similar fashion as CsA, and that silencing of CsCyp, as well as treatments with CsA, enhance canker lesions in X. citri-infected leaves. Given that CsCyp appears to function as a negative regulator of cell growth and that Ess1 negatively regulates transcription elongation in yeast, we propose that PthAs activate host transcription by inhibiting the PPIase activity of CsCyp on the CTD.

  18. Identifying a novel role for X-prolyl aminopeptidase (Xpnpep) 2 in CrVI-induced adverse effects on germ cell nest breakdown and follicle development in rats.

    Science.gov (United States)

    Banu, Sakhila K; Stanley, Jone A; Sivakumar, Kirthiram K; Arosh, Joe A; Barhoumi, Rola; Burghardt, Robert C

    2015-03-01

    Environmental exposure to endocrine-disrupting chemicals (EDCs) is one cause of premature ovarian failure (POF). Hexavalent chromium (CrVI) is a heavy metal EDC widely used in more than 50 industries, including chrome plating, welding, wood processing, and tanneries. Recent data from U.S. Environmental Protection Agency indicate increased levels of Cr in drinking water from several American cities, which potentially predispose residents to various health problems. Recently, we demonstrated that gestational exposure to CrVI caused POF in F1 offspring. The current study was performed to identify the molecular mechanism behind CrVI-induced POF. Pregnant rats were treated with 25 ppm of potassium dichromate from Gestational Day (GD) 9.5 to GD 14.5 through drinking water, and the fetuses were exposed to CrVI through transplacental transfer. Ovaries were removed from the fetuses or pups on Embryonic Day (ED) 15.5, ED 17.5, Postnatal Day (PND) 1, PND 4, or PND 25, and various analyses were performed. Results showed that gestational exposure to CrVI: 1) increased germ cell/oocyte apoptosis and advanced germ cell nest (GCN) breakdown; 2) increased X-prolyl aminopeptidase (Xpnpep) 2, a POF marker in humans, during GCN breakdown; 3) decreased Xpnpep2 during postnatal follicle development; and 4) increased colocalization of Xpnpep2 with Col3 and Col4. We also found that Xpnpep2 inversely regulated the expression of Col1, Col3, and Col4 in all the developmental stages studied. Thus, CrVI advanced GCN breakdown and increased follicle atresia in F1 female progeny by targeting Xpnpep2. © 2015 by the Society for the Study of Reproduction, Inc.

  19. Therapeutic Potential of a Prolyl Hydroxylase Inhibitor FG-4592 for Parkinson’s Diseases in Vitro and in Vivo: Regulation of Redox Biology and Mitochondrial Function

    Directory of Open Access Journals (Sweden)

    Xuan Li

    2018-04-01

    Full Text Available As the main transcription factor that regulates the cellular responses to hypoxia, Hypoxia-inducible factor-1α (HIF-1α plays an important role in the pathogenesis of Parkinson’s disease (PD. HIF-1α is normally degraded through ubiquitination after hydroxylation by prolyl hydroxylases (PHD. Emerging evidence has suggested that HIF PHD inhibitors (HIF-PHI may have neuroprotective effects on PD through increasing HIF-1α levels. However, the therapeutic benefit of HIF-PHI for PD remains poorly explored due to the lack of proper clinical compounds and understanding of the underlying molecular mechanisms. In this study, we examined the therapeutic benefit of a new HIF-PHI, FG-4592, which is currently in phase 3 clinical trials to treat anemia in patients with chronic kidney diseases (CKD in PD models. FG-4592 attenuates MPP+ -induced apoptosis and loss of tyrosine hydroxylase (TH in SH-SY5Y cells. Pretreatment with FG-4592 mitigates MPP+-induced loss of mitochondrial membrane potential (MMP, mitochondrial oxygen consumption rate (OCR, production of reactive oxygen species (ROS and ATP. Furthermore, FG-4592 counterbalances the oxidative stress through up-regulating nuclear factor erythroid 2 p45-related factor 2 (Nrf-2, heme oxygenase-1 (HO-1 and superoxide dismutase 2 (SOD2. FG-4592 treatment also induces the expression of Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α through increasing the phosphorylation of AMP-activated protein kinase (AMPK. In MPTP-treated mice, FG-4592 protects against MPTP-induced loss of TH-positive neurons of substantia nigra and attenuates behavioral impairments. Collectively, our study demonstrates that FG-4592 is a promising therapeutic strategy for PD through improving the mitochondrial function under oxidative stress.

  20. Modulation of neutrophil superoxide generation by inhibitors of protein kinase C, calmodulin, diacylglycerol and myosin light chain kinases, and peptidyl prolyl cis-trans isomerase.

    Science.gov (United States)

    Bergstrand, H; Eriksson, T; Hallberg, A; Johansson, B; Karabelas, K; Michelsen, P; Nybom, A

    1992-12-01

    To assess the role of protein kinase C (PKC) in the respiratory burst of adherent human polymorphonuclear leukocytes (PMNL), reduction of ferricytochrome C by cells triggered with a phorbol ester (PMA), ionophore A23187, serum-treated zymosan (STZ) or three lipid derivatives, 3-decanoyl-sn-glycerol (G-3-OCOC9), (R,R)-1,4-diethyl-2-O-decyl-L-tartrate (Tt-2-OC10) and 3-decyloxy-5-hydroxymethylphenol (DHP) was examined in a microtiter plate procedure in the presence of inhibitors of PKC and, for comparison, inhibitors of calmodulin, diacylglycerol and myosin light chain kinases and the peptidyl-prolyl cis-trans isomerase activity of fujiphilin. 1) Of the protein kinase inhibitors examined, Ro 31-7549 and staurosporine reduced responses to all stimuli except possibly STZ; in contrast, K252a and the myosin light chain kinase inhibitors ML-7 and ML-9 blocked responses to A23187 and STZ better than those triggered by PMA. H-7 reduced responses to A23187, DHP and G-3-OCOC9, and calphostin, palmitoyl carnitine, sphingosine and the multifunctional drugs TMB-8 and W-7 reduced A23187; they also, when examined, reduced decane derivative-induced O2- production more effectively than PMA- and STZ-triggered responses. Polymyxin B, 4 alpha-PMA and retinal displayed no inhibitory capacity. 2) Of the selective calmodulin antagonists, CGS 9343B, Ro 22-4839 and calmidazolium did not inhibit the oxidative response irrespective of the stimulus used, whereas metofenazate reduced those evoked by A23187, DHP, G-3-OCOC9 and STZ.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. First-in-man-proof of concept study with molidustat: a novel selective oral HIF-prolyl hydroxylase inhibitor for the treatment of renal anaemia.

    Science.gov (United States)

    Böttcher, M; Lentini, S; Arens, E R; Kaiser, A; van der Mey, D; Thuss, U; Kubitza, D; Wensing, G

    2018-07-01

    Insufficient erythropoietin (EPO) synthesis is a relevant cause of renal anaemia in patients with chronic kidney disease. Molidustat, a selective hypoxia-inducible factor prolyl hydroxylase (HIF-PH) inhibitor, increases endogenous EPO levels dose dependently in preclinical models. We examined the pharmacokinetics, safety, tolerability and effect on EPO levels of single oral doses of molidustat in healthy male volunteers. This was a single-centre, randomized, single-blind, placebo-controlled, group-comparison, dose-escalation study. Molidustat was administered at doses of 5, 12.5, 25, 37.5 or 50 mg as a polyethylene glycol-based solution. In total, 45 volunteers received molidustat and 14 received placebo. Molidustat was absorbed rapidly, and the mean maximum plasma concentration and area under the concentration-time curve increased dose dependently. The mean terminal half-life was 4.64-10.40 h. A significant increase in endogenous EPO was observed following single oral doses of molidustat of 12.5 mg and above. Geometric mean peak EPO levels were 14.8 IU l -1 (90% confidence interval 13.0, 16.9) for volunteers who received placebo and 39.8 IU l -1 (90% confidence interval: 29.4, 53.8) for those who received molidustat 50 mg. The time course of EPO levels resembled the normal diurnal variation in EPO. Maximum EPO levels were observed approximately 12 h postdose and returned to baseline after approximately 24-48 h. All doses of molidustat were well tolerated and there were no significant changes in vital signs or laboratory safety parameters. Oral administration of molidustat to healthy volunteers elicited a dose-dependent increase in endogenous EPO. These results support the ongoing development of molidustat as a potential new treatment for patients with renal anaemia. © 2018 The British Pharmacological Society.

  2. Induction of erythropoiesis by hypoxia-inducible factor prolyl hydroxylase inhibitors without promotion of tumor initiation, progression, or metastasis in a VEGF-sensitive model of spontaneous breast cancer

    Directory of Open Access Journals (Sweden)

    Seeley TW

    2017-03-01

    Full Text Available Todd W Seeley, Mark D Sternlicht, Stephen J Klaus, Thomas B Neff, David Y Liu Therapeutics R&D, FibroGen, Inc., San Francisco, CA, USA Abstract: The effects of pharmacological hypoxia-inducible factor (HIF stabilization were investigated in the MMTV-Neundl-YD5 (NeuYD mouse model of breast cancer. This study first confirmed the sensitivity of this model to increased vascular endothelial growth factor (VEGF, using bigenic NeuYD;MMTV-VEGF-25 mice. Tumor initiation was dramatically accelerated in bigenic animals. Bigenic tumors were also more aggressive, with shortened doubling times and increased lung metastasis as compared to NeuYD controls. In separate studies, NeuYD mice were treated three times weekly from 7 weeks of age until study end with two different HIF prolyl hydroxylase inhibitors (HIF-PHIs, FG-4497 or roxadustat (FG-4592. In NeuYD mice, HIF-PHI treatments elevated erythropoiesis markers, but no differences were detected in tumor onset or the phenotypes of established tumors. Keywords: cancer progression, erythropoiesis, hypoxia-inducible factor, hypoxia-inducible factor prolyl hydroxylase inhibitors, vascular endothelial growth factor, MMTV-Neu breast cancer model

  3. Solid substrate fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Tengerdy, R P

    1985-04-01

    Solid Substrate Fermentation (SSF) describes the microbiological tranformation of biological materials in their natural state, in contrast with liquid or submerged fermentations which are carried out in dilute solutions or slurries. The most important industrial microorganisms used in SSF are filamentous fungi and the critical factors in their growth are the control of the moisture level and the temperature. Traditionally, most SSFs are conducted in shallow trays (so that heat build up is avoided) and stacked in a moist chamber, however, the modern SSF should be able to mix large amounts of substrate for a uniform fermentation, maximum automization scale-up of the process, continuous operation and fermentation control and a promising new design is the Helical screw fermenter. At the present time SSF is used in the production of foods (e.g. mushrooms and oriental foods) in municipal, agricultural and industrial solid waste disposal and in the production of enzymes and speciality chemicals but it does not seem likely that it will replace prevalent liquid fermentation technologies. 29 references.

  4. Maintainable substrate carrier for electroplating

    Science.gov (United States)

    Chen, Chen-An [Milpitas, CA; Abas, Emmanuel Chua [Laguna, PH; Divino, Edmundo Anida [Cavite, PH; Ermita, Jake Randal G [Laguna, PH; Capulong, Jose Francisco S [Laguna, PH; Castillo, Arnold Villamor [Batangas, PH; Ma,; Xiaobing, Diana [Saratoga, CA

    2012-07-17

    One embodiment relates to a substrate carrier for use in electroplating a plurality of substrates. The carrier includes a non-conductive carrier body on which the substrates are placed and conductive lines embedded within the carrier body. A plurality of conductive clip attachment parts are attached in a permanent manner to the conductive lines embedded within the carrier body. A plurality of contact clips are attached in a removable manner to the clip attachment parts. The contact clips hold the substrates in place and conductively connecting the substrates with the conductive lines. Other embodiments, aspects and features are also disclosed.

  5. Expression and DNA methylation levels of prolyl hydroxylases PHD1, PHD2, PHD3 and asparaginyl hydroxylase FIH in colorectal cancer

    International Nuclear Information System (INIS)

    Rawluszko, Agnieszka A; Bujnicka, Katarzyna E; Horbacka, Karolina; Krokowicz, Piotr; Jagodziński, Paweł P

    2013-01-01

    Colorectal cancer (CRC) is one of the most common and comprehensively studied malignancies. Hypoxic conditions during formation of CRC may support the development of more aggressive cancers. Hypoxia inducible factor (HIF), a major player in cancerous tissue adaptation to hypoxia, is negatively regulated by the family of prolyl hydroxylase enzymes (PHD1, PHD2, PHD3) and asparaginyl hydroxylase, called factor inhibiting HIF (FIH). PHD1, PHD2, PHD3 and FIH gene expression was evaluated using quantitative RT-PCR and western blotting in primary colonic adenocarcinoma and adjacent histopathologically unchanged colonic mucosa from patients who underwent radical surgical resection of the colon (n = 90), and the same methods were used for assessment of PHD3 gene expression in HCT116 and DLD-1 CRC cell lines. DNA methylation levels of the CpG island in the promoter regulatory region of PHD1, PHD2, PHD3 and FIH were assessed using bisulfite DNA sequencing and high resolution melting analysis (HRM) for patients and HRM analysis for CRC cell lines. We found significantly lower levels of PHD1, PHD2 and PHD3 transcripts (p = 0.00026; p < 0.00001; p < 0.00001) and proteins (p = 0.004164; p = 0.0071; p < 0.00001) in primary cancerous than in histopathologically unchanged tissues. Despite this, we did not observe statistically significant differences in FIH transcript levels between cancerous and histopathologically unchanged colorectal tissue, but we found a significantly increased level of FIH protein in CRC (p = 0.0169). The reduced PHD3 expression was correlated with significantly increased DNA methylation in the CpG island of the PHD3 promoter regulatory region (p < 0.0001). We did not observe DNA methylation in the CpG island of the PHD1, PHD2 or FIH promoter in cancerous and histopathologically unchanged colorectal tissue. We also showed that 5-Aza-2’-deoxycytidine induced DNA demethylation leading to increased PHD3 transcript and protein level in HCT116 cells. We

  6. An Intracellular Peptidyl-Prolyl cis/trans Isomerase Is Required for Folding and Activity of the Staphylococcus aureus Secreted Virulence Factor Nuclease.

    Science.gov (United States)

    Wiemels, Richard E; Cech, Stephanie M; Meyer, Nikki M; Burke, Caleb A; Weiss, Andy; Parks, Anastacia R; Shaw, Lindsey N; Carroll, Ronan K

    2017-01-01

    Staphylococcus aureus is an important human pathogen that relies on a large repertoire of secreted and cell wall-associated proteins for pathogenesis. Consequently, the ability of the organism to cause disease is absolutely dependent on its ability to synthesize and successfully secrete these proteins. In this study, we investigate the role of peptidyl-prolyl cis/trans isomerases (PPIases) on the activity of the S. aureus secreted virulence factor nuclease (Nuc). We identify a staphylococcal cyclophilin-type PPIase (PpiB) that is required for optimal activity of Nuc. Disruption of ppiB results in decreased nuclease activity in culture supernatants; however, the levels of Nuc protein are not altered, suggesting that the decrease in activity results from misfolding of Nuc in the absence of PpiB. We go on to demonstrate that PpiB exhibits PPIase activity in vitro, is localized to the bacterial cytosol, and directly interacts with Nuc in vitro to accelerate the rate of Nuc refolding. Finally, we demonstrate an additional role for PpiB in S. aureus hemolysis and demonstrate that the S. aureus parvulin-type PPIase PrsA also plays a role in the activity of secreted virulence factors. The deletion of prsA leads to a decrease in secreted protease and phospholipase activity, similar to that observed in other Gram-positive pathogens. Together, these results demonstrate, for the first time to our knowledge, that PPIases play an important role in the secretion of virulence factors in S. aureus IMPORTANCE: Staphylococcus aureus is a highly dangerous bacterial pathogen capable of causing a variety of infections throughout the human body. The ability of S. aureus to cause disease is largely due to an extensive repertoire of secreted and cell wall-associated proteins, including adhesins, toxins, exoenzymes, and superantigens. These virulence factors, once produced, are typically transported across the cell membrane by the secretory (Sec) system in a denatured state. Consequently

  7. Severe Extracellular Matrix Abnormalities and Chondrodysplasia in Mice Lacking Collagen Prolyl 4-Hydroxylase Isoenzyme II in Combination with a Reduced Amount of Isoenzyme I.

    Science.gov (United States)

    Aro, Ellinoora; Salo, Antti M; Khatri, Richa; Finnilä, Mikko; Miinalainen, Ilkka; Sormunen, Raija; Pakkanen, Outi; Holster, Tiina; Soininen, Raija; Prein, Carina; Clausen-Schaumann, Hauke; Aszódi, Attila; Tuukkanen, Juha; Kivirikko, Kari I; Schipani, Ernestina; Myllyharju, Johanna

    2015-07-03

    Collagen prolyl 4-hydroxylases (C-P4H-I, C-P4H-II, and C-P4H-III) catalyze formation of 4-hydroxyproline residues required to form triple-helical collagen molecules. Vertebrate C-P4Hs are α2β2 tetramers differing in their catalytic α subunits. C-P4H-I is the major isoenzyme in most cells, and inactivation of its catalytic subunit (P4ha1(-/-)) leads to embryonic lethality in mouse, whereas P4ha1(+/-) mice have no abnormalities. To study the role of C-P4H-II, which predominates in chondrocytes, we generated P4ha2(-/-) mice. Surprisingly, they had no apparent phenotypic abnormalities. To assess possible functional complementarity, we established P4ha1(+/-);P4ha2(-/-) mice. They were smaller than their littermates, had moderate chondrodysplasia, and developed kyphosis. A transient inner cell death phenotype was detected in their developing growth plates. The columnar arrangement of proliferative chondrocytes was impaired, the amount of 4-hydroxyproline and the Tm of collagen II were reduced, and the extracellular matrix was softer in the growth plates of newborn P4ha1(+/-);P4ha2(-/-) mice. No signs of uncompensated ER stress were detected in the mutant growth plate chondrocytes. Some of these defects were also found in P4ha2(-/-) mice, although in a much milder form. Our data show that C-P4H-I can to a large extent compensate for the lack of C-P4H-II in proper endochondral bone development, but their combined partial and complete inactivation, respectively, leads to biomechanically impaired extracellular matrix, moderate chondrodysplasia, and kyphosis. Our mouse data suggest that inactivating mutations in human P4HA2 are not likely to lead to skeletal disorders, and a simultaneous decrease in P4HA1 function would most probably be required to generate such a disease phenotype. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Autophagy regulated by prolyl isomerase Pin1 and phospho-Ser-GSK3αβ involved in protection of oral squamous cell carcinoma against cadmium toxicity

    Energy Technology Data Exchange (ETDEWEB)

    So, Keum-Young [Department of Anesthesiology and Pain Medicine College of Dentistry, Chosun University, 309 Pilmundaero, Dong-gu, Gwangju 501-759 (Korea, Republic of); Ahn, Sang-Gun [Department of Pathology, College of Dentistry, Chosun University, 309 Pilmundaero, Dong-gu, Gwangju 501-759 (Korea, Republic of); Oh, Seon-Hee, E-mail: seonh@chosun.ac.kr [Department of Premedicine, School of Medicine, College of Dentistry, Chosun University, 309 Pilmundaero, Dong-gu, Gwangju 501-759 (Korea, Republic of)

    2015-10-23

    Prolyl isomerase Pin1 plays an important role in cell proliferation and is overexpressed in many human tumors. However, its role in autophagy induction remains undefined. Here we show that Pin1 regulates cell survival via autophagy in cadmium (Cd)-exposed oral squamous cell carcinoma (OSCC). OSCC exposure to Cd induced autophagy, as demonstrated by the formation of green fluorescent punctae in transfected cells expressing GFP-conjugated microtubule-associated protein light chain 3 (LC3) and by LC3 flux in the presence of autophagy inhibitors. Suppression of Atg5 enhanced Cd-induced apoptosis, indicating that autophagy is involved in cell protection. In dose–response experiments, cleavage of procaspase-3, PARP-1, and LC3-II was induced by Cd with an IC{sub 50} of 45 μM. Expression of Pin1 was decreased at or above the Cd IC{sub 50} value and was inversely correlated with the level of phospho(p)-Ser-GSK3αβ. Genetic or pharmacologic inhibition of Pin1 suppressed Cd-induced autophagy, but increased p-Akt-mediated p-Ser-GSK3αβ; this was reversed by overexpression of Pin1. However, suppression of GSK3αβ inhibited Cd-induced autophagy and induced apoptosis, which could be reversed by overexpression of GSK3β. The PI3K inhibitor Ly294002 blocked p-Akt-mediated increases in p-Ser-GSK3αβ and autophagy and induced apoptosis. Therefore, p-Ser-GSK3αβ can directly regulate Cd-induced autophagy, although its function is suppressed by Pin1. Collectively, the present results indicate that targeting Pin1 and GSK3αβ at the same time could be an effective therapeutic tool for Cd-induced carcinogenesis. - Highlights: • Pin1 regulated autophagy to protect cells from cadmium toxicity. • Pin1 suppression inhibited cadmium-induced autophagy and induced apoptosis. • Pin1 inhibited the function of p-Ser-GSK3αβ in autophagy regulation. • p-Ser-GSK3αβ regulated autophagy independently of Pin1.

  9. Autophagy regulated by prolyl isomerase Pin1 and phospho-Ser-GSK3αβ involved in protection of oral squamous cell carcinoma against cadmium toxicity

    International Nuclear Information System (INIS)

    So, Keum-Young; Ahn, Sang-Gun; Oh, Seon-Hee

    2015-01-01

    Prolyl isomerase Pin1 plays an important role in cell proliferation and is overexpressed in many human tumors. However, its role in autophagy induction remains undefined. Here we show that Pin1 regulates cell survival via autophagy in cadmium (Cd)-exposed oral squamous cell carcinoma (OSCC). OSCC exposure to Cd induced autophagy, as demonstrated by the formation of green fluorescent punctae in transfected cells expressing GFP-conjugated microtubule-associated protein light chain 3 (LC3) and by LC3 flux in the presence of autophagy inhibitors. Suppression of Atg5 enhanced Cd-induced apoptosis, indicating that autophagy is involved in cell protection. In dose–response experiments, cleavage of procaspase-3, PARP-1, and LC3-II was induced by Cd with an IC_5_0 of 45 μM. Expression of Pin1 was decreased at or above the Cd IC_5_0 value and was inversely correlated with the level of phospho(p)-Ser-GSK3αβ. Genetic or pharmacologic inhibition of Pin1 suppressed Cd-induced autophagy, but increased p-Akt-mediated p-Ser-GSK3αβ; this was reversed by overexpression of Pin1. However, suppression of GSK3αβ inhibited Cd-induced autophagy and induced apoptosis, which could be reversed by overexpression of GSK3β. The PI3K inhibitor Ly294002 blocked p-Akt-mediated increases in p-Ser-GSK3αβ and autophagy and induced apoptosis. Therefore, p-Ser-GSK3αβ can directly regulate Cd-induced autophagy, although its function is suppressed by Pin1. Collectively, the present results indicate that targeting Pin1 and GSK3αβ at the same time could be an effective therapeutic tool for Cd-induced carcinogenesis. - Highlights: • Pin1 regulated autophagy to protect cells from cadmium toxicity. • Pin1 suppression inhibited cadmium-induced autophagy and induced apoptosis. • Pin1 inhibited the function of p-Ser-GSK3αβ in autophagy regulation. • p-Ser-GSK3αβ regulated autophagy independently of Pin1.

  10. MiR-30e suppresses proliferation of hepatoma cells via targeting prolyl 4-hydroxylase subunit alpha-1 (P4HA1) mRNA

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Guoxing [State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin (China); Shi, Hui [State Key Laboratory of Medicinal Chemical Biology, Department of Biochemistry, College of Life Sciences, Nankai University, Tianjin (China); Li, Jiong; Yang, Zhe [State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin (China); Fang, Runping; Ye, Lihong [State Key Laboratory of Medicinal Chemical Biology, Department of Biochemistry, College of Life Sciences, Nankai University, Tianjin (China); Zhang, Weiying, E-mail: zhwybao@nankai.edu.cn [State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin (China); Zhang, Xiaodong, E-mail: zhangxd@nankai.edu.cn [State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin (China)

    2016-04-08

    Aberrant microRNA expression has been shown to be characteristic of many cancers. It has been reported that the expression levels of miR-30e are decreased in liver cancer tissues. However, the role of miR-30e in hepatocellular carcinoma remains poorly understood. In the present study, we investigated the significance of miR-30e in hepatocarcinogenesis. Bioinformatics analysis reveals a putative target site of miR-30e in the 3′-untranslated region (3′UTR) of prolyl 4-hydroxylase subunit alpha-1 (P4HA1) mRNA. Moreover, luciferase reporter gene assays verified that miR-30e directly targeted 3′UTR of P4HA1 mRNA. Then, we demonstrated that miR-30e was able to reduce the expression of P4HA1 at the levels of mRNA and protein using reverse transcription-polymerase chain reaction and Western blot analysis. Enforced expression of miR-30e suppressed proliferation of HepG2 cells by 5-ethynyl-2-deoxyuridine (EdU) assay and reduced colony formation of these cells by colony formation analysis. Conversely, anti-miR-30e enhanced the proliferation of hepatoma cells in vitro. Interestingly, the ectopic expression of P4HA1 could efficiently rescue the inhibition of cell proliferation mediated by miR-30e in HepG2 cells. Meanwhile, silencing of P4HA1 abolished the anti-miR-30e-induced proliferation of cells. Clinically, quantitative real-time PCR showed that miR-30e was down-regulated in liver tumor tissues relative to their peritumor tissues. The expression levels of miR-30e were negatively correlated to those of P4HA1 mRNA in clinical liver tumor tissues. Thus, we conclude that miR-30e suppresses proliferation of hepatoma cells through targeting P4HA1 mRNA. Our finding provides new insights into the mechanism of hepatocarcinogenesis. - Highlights: • P4HA1 is a novel target gene of miR-30e. • P4HA1 is increased in clinical HCC tissues. • MiR-30e is negatively correlated with P4HA1 in clinical HCC tissues. • MiR-30e suppresses the proliferation of HCC cells through

  11. Hydroxylation of recombinant human collagen type I alpha 1 in transgenic maize co-expressed with a recombinant human prolyl 4-hydroxylase

    Directory of Open Access Journals (Sweden)

    Pappu Kameshwari M

    2011-06-01

    Full Text Available Abstract Background Collagens require the hydroxylation of proline (Pro residues in their triple-helical domain repeating sequence Xaa-Pro-Gly to function properly as a main structural component of the extracellular matrix in animals at physiologically relevant conditions. The regioselective proline hydroxylation is catalyzed by a specific prolyl 4-hydroxylase (P4H as a posttranslational processing step. Results A recombinant human collagen type I α-1 (rCIα1 with high percentage of hydroxylated prolines (Hyp was produced in transgenic maize seeds when co-expressed with both the α- and β- subunits of a recombinant human P4H (rP4H. Germ-specific expression of rCIα1 using maize globulin-1 gene promoter resulted in an average yield of 12 mg/kg seed for the full-length rCIα1 in seeds without co-expression of rP4H and 4 mg/kg seed for the rCIα1 (rCIα1-OH in seeds with co-expression of rP4H. High-resolution mass spectrometry (HRMS analysis revealed that nearly half of the collagenous repeating triplets in rCIα1 isolated from rP4H co-expressing maize line had the Pro residues changed to Hyp residues. The HRMS analysis determined the Hyp content of maize-derived rCIα1-OH as 18.11%, which is comparable to the Hyp level of yeast-derived rCIα1-OH (17.47% and the native human CIa1 (14.59%, respectively. The increased Hyp percentage was correlated with a markedly enhanced thermal stability of maize-derived rCIα1-OH when compared to the non-hydroxylated rCIα1. Conclusions This work shows that maize has potential to produce adequately modified exogenous proteins with mammalian-like post-translational modifications that may be require for their use as pharmaceutical and industrial products.

  12. PLZT capacitor on glass substrate

    Science.gov (United States)

    Fairchild, M. Ray; Taylor, Ralph S.; Berlin, Carl W.; Wong, Celine W. K.; Ma, Beihai; Balachandran, Uthamalingam

    2016-01-05

    A lead-lanthanum-zirconium-titanate (PLZT) capacitor on a substrate formed of glass. The first metallization layer is deposited on a top side of the substrate to form a first electrode. The dielectric layer of PLZT is deposited over the first metallization layer. The second metallization layer deposited over the dielectric layer to form a second electrode. The glass substrate is advantageous as glass is compatible with an annealing process used to form the capacitor.

  13. Sealed substrate carrier for electroplating

    Science.gov (United States)

    Ganti, Kalyana Bhargava [Fremont, CA

    2012-07-17

    One embodiment relates to a substrate carrier for use in electroplating a plurality of substrates. The substrate carrier includes a non-conductive carrier body on which the substrates are held, and conductive lines are embedded within the carrier body. A conductive bus bar is embedded into a top side of the carrier body and is conductively coupled to the conductive lines. A thermoplastic overmold covers a portion of the bus bar, and there is a plastic-to-plastic bond between the thermoplastic overmold and the non-conductive carrier body. Other embodiments, aspects and features are also disclosed.

  14. Induction of erythropoiesis by hypoxia-inducible factor prolyl hydroxylase inhibitors without promotion of tumor initiation, progression, or metastasis in a VEGF-sensitive model of spontaneous breast cancer

    Science.gov (United States)

    Seeley, Todd W; Sternlicht, Mark D; Klaus, Stephen J; Neff, Thomas B; Liu, David Y

    2017-01-01

    The effects of pharmacological hypoxia-inducible factor (HIF) stabilization were investigated in the MMTV-Neundl-YD5 (NeuYD) mouse model of breast cancer. This study first confirmed the sensitivity of this model to increased vascular endothelial growth factor (VEGF), using bigenic NeuYD;MMTV-VEGF-25 mice. Tumor initiation was dramatically accelerated in bigenic animals. Bigenic tumors were also more aggressive, with shortened doubling times and increased lung metastasis as compared to NeuYD controls. In separate studies, NeuYD mice were treated three times weekly from 7 weeks of age until study end with two different HIF prolyl hydroxylase inhibitors (HIF-PHIs), FG-4497 or roxadustat (FG-4592). In NeuYD mice, HIF-PHI treatments elevated erythropoiesis markers, but no differences were detected in tumor onset or the phenotypes of established tumors. PMID:28331872

  15. Insight into a conformation of the PNA-PNA duplex with (2‧R,4‧R)- and (2‧R,4‧S)-prolyl-(1S,2S)-2-aminocyclopentanecarboxylic acid backbones

    Science.gov (United States)

    Maitarad, Amphawan; Poomsuk, Nattawee; Vilaivan, Chotima; Vilaivan, Tirayut; Siriwong, Khatcharin

    2018-04-01

    Suitable conformations for peptide nucleic acid (PNA) self-hybrids with (2‧R,4‧R)- and (2‧R,4‧S)-prolyl-(1S,2S)-2-aminocyclopentanecarboxylic acid backbones (namely, acpcPNA and epi-acpcPNA, respectively) were investigated based on molecular dynamics simulations. The results revealed that hybridization of the acpcPNA was observed only in the parallel direction, with a conformation close to the P-type structure. In contrast, self-hybrids of the epi-acpcPNA were formed in the antiparallel and parallel directions; the antiparallel duplex adopted the B-form conformation, and the parallel duplex was between B- and P-forms. The calculated binding energies and the experimental data indicate that the antiparallel epi-acpcPNA self-hybrid was more stable than the parallel duplex.

  16. Receptor type I and type II binding regions and the peptidyl-prolyl isomerase site of cyclophilin B are required for enhancement of T-lymphocyte adhesion to fibronectin.

    Science.gov (United States)

    Carpentier, Mathieu; Allain, Fabrice; Slomianny, Marie-Christine; Durieux, Sandrine; Vanpouille, Christophe; Haendler, Bernard; Spik, Geneviève

    2002-04-23

    Cyclophilin B (CyPB), a cyclosporin A (CsA) binding protein, interacts with two types of binding sites at the surface of T-lymphocytes. The type I sites correspond to functional receptors involved in endocytosis and the type II sites to sulfated glycosaminoglycans (GAGs). Mutational analysis of CyPB has revealed that W128, which is part of the CsA-binding pocket, is implicated in the binding to the functional type I receptors and that two amino acid clusters located in the N-terminus ensure the binding to GAGs. The peptidyl-prolyl isomerase activity of CyPB is not required for receptor binding. We have recently demonstrated that CyPB enhances adhesion of peripheral blood T-lymphocytes to fibronectin, a component of the extracellular matrix. We intended to identify additional amino acids involved in the binding of CyPB to its functional type I receptor and to determine regions responsible for the stimulation of peripheral blood T-lymphocyte adhesion. We determined that residues R76, G77, K132, D155, and D158 of the calcineurin (CN) interacting region were implicated in the recognition of type I receptor but not of GAGs. We also found that two different changes in the N-terminal extension that abated binding to GAGs prevented adhesion of peripheral blood T-lymphocytes to coated CyPB, whereas abbrogation of the PPIase activity had no effect. On the other hand, the adhesion of peripheral blood T-lymphocytes to coated fibronectin was not stimulated by CyPB mutants devoid of either type I receptor or GAGs binding activity or by mutants of the PPIase site. Altogether, the results demonstrate that different regions of CyPB are involved in peripheral blood T-lymphocyte activation and imply a novel important physiological function for peptidyl-prolyl isomerase activity.

  17. Direct cooled power electronics substrate

    Science.gov (United States)

    Wiles, Randy H [Powell, TN; Wereszczak, Andrew A [Oak Ridge, TN; Ayers, Curtis W [Kingston, TN; Lowe, Kirk T [Knoxville, TN

    2010-09-14

    The disclosure describes directly cooling a three-dimensional, direct metallization (DM) layer in a power electronics device. To enable sufficient cooling, coolant flow channels are formed within the ceramic substrate. The direct metallization layer (typically copper) may be bonded to the ceramic substrate, and semiconductor chips (such as IGBT and diodes) may be soldered or sintered onto the direct metallization layer to form a power electronics module. Multiple modules may be attached to cooling headers that provide in-flow and out-flow of coolant through the channels in the ceramic substrate. The modules and cooling header assembly are preferably sized to fit inside the core of a toroidal shaped capacitor.

  18. Dielectric coatings on metal substrates

    International Nuclear Information System (INIS)

    Glaros, S.S.; Baker, P.; Milam, D.

    1976-01-01

    Large aperture, beryllium substrate-based mirrors have been used to focus high intensity pulsed laser beams. Finished surfaces have high reflectivity, low wavefront distortion, and high laser damage thresholds. This paper describes the development of a series of metallic coatings, surface finishing techniques, and dielectric overcoatings to meet specified performance requirements. Beryllium substrates were coated with copper, diamond-machined to within 5 micro-inches to final contour, nickel plated, and abrasively figured to final contour. Bond strengths for several bonding processes are presented. Dielectric overcoatings were deposited on finished multimetallic substrates to increase both reflectivity and the damage thresholds. Coatings were deposited using both high and low temperature processes which induce varying stresses in the finished coating substrate system. Data are presented to show the evolution of wavefront distortion, reflectivity, and damage thresholds throughout the many steps involved in fabrication

  19. Probing protein phosphatase substrate binding

    DEFF Research Database (Denmark)

    Højlys-Larsen, Kim B.; Sørensen, Kasper Kildegaard; Jensen, Knud Jørgen

    2012-01-01

    Proteomics and high throughput analysis for systems biology can benefit significantly from solid-phase chemical tools for affinity pull-down of proteins from complex mixtures. Here we report the application of solid-phase synthesis of phosphopeptides for pull-down and analysis of the affinity...... profile of the integrin-linked kinase associated phosphatase (ILKAP), a member of the protein phosphatase 2C (PP2C) family. Phosphatases can potentially dephosphorylate these phosphopeptide substrates but, interestingly, performing the binding studies at 4 °C allowed efficient binding to phosphopeptides......, without the need for phosphopeptide mimics or phosphatase inhibitors. As no proven ILKAP substrates were available, we selected phosphopeptide substrates among known PP2Cδ substrates including the protein kinases: p38, ATM, Chk1, Chk2 and RSK2 and synthesized directly on PEGA solid supports through a BAL...

  20. Graphene on insulating crystalline substrates

    International Nuclear Information System (INIS)

    Akcoeltekin, S; El Kharrazi, M; Koehler, B; Lorke, A; Schleberger, M

    2009-01-01

    We show that it is possible to prepare and identify ultra-thin sheets of graphene on crystalline substrates such as SrTiO 3 , TiO 2 , Al 2 O 3 and CaF 2 by standard techniques (mechanical exfoliation, optical and atomic force microscopy). On the substrates under consideration we find a similar distribution of single layer, bilayer and few-layer graphene and graphite flakes as with conventional SiO 2 substrates. The optical contrast C of a single graphene layer on any of those substrates is determined by calculating the optical properties of a two-dimensional metallic sheet on the surface of a dielectric, which yields values between C = -1.5% (G/TiO 2 ) and C = -8.8% (G/CaF 2 ). This contrast is in reasonable agreement with experimental data and is sufficient to make identification by an optical microscope possible. The graphene layers cover the crystalline substrate in a carpet-like mode and the height of single layer graphene on any of the crystalline substrates as determined by atomic force microscopy is d SLG = 0.34 nm and thus much smaller than on SiO 2 .

  1. Methods of etching a substrate

    Energy Technology Data Exchange (ETDEWEB)

    Cosmo, J J; Gambino, R J; Harper, J M.E.

    1979-05-16

    The invention relates to a method of etching a substrate. The substrate is located opposite a target electrode in a vacuum chamber, and the surface of the target electrode is bombarded with energetic particles of atomic dimensions. The target electrode is an intermetallic composition (compound, alloy or finely divided homogeneous mixture) of two metals A and B such that upon bombardment the electrode emits negative ions of metal B which have sufficient energy to produce etching of the substrate. Many target materials are exemplified. Typically the metal A has an electronegativity XA and metal B has an electronegativity XB such that Xb - Xa is greater than about 2.55 electron volts, with the exception of combinations of metals having a fractional ionicity Q less than about 0.314. The source of the energetic particles may be an ionised gas in the vacuum chamber. The apparatus and its mode of operation are described in detail.

  2. Methods of etching a substrate

    International Nuclear Information System (INIS)

    Cosmo, J.J.; Gambino, R.J.; Harper, J.M.E.

    1979-01-01

    The invention relates to a method of etching a substrate. The substrate is located opposite a target electrode in a vacuum chamber, and the surface of the target electrode is bombarded with energetic particles of atomic dimensions. The target electrode is an intermetallic composition (compound, alloy or finely divided homogeneous mixture) of two metals A and B such that upon bombardment the electrode emits negative ions of metal B which have sufficient energy to produce etching of the substrate. Many target materials are exemplified. Typically the metal A has an electronegativity XA and metal B has an electronegativity XB such that Xb - Xa is greater than about 2.55 electron volts, with the exception of combinations of metals having a fractional ionicity Q less than about 0.314. The source of the energetic particles may be an ionised gas in the vacuum chamber. The apparatus and its mode of operation are described in detail. (U.K.)

  3. Porous substrates filled with nanomaterials

    Science.gov (United States)

    Worsley, Marcus A.; Baumann, Theodore F.; Satcher, Jr., Joe H.; Stadermann, Michael

    2018-04-03

    A composition comprising: at least one porous carbon monolith, such as a carbon aerogel, comprising internal pores, and at least one nanomaterial, such as carbon nanotubes, disposed uniformly throughout the internal pores. The nanomaterial can be disposed in the middle of the monolith. In addition, a method for making a monolithic solid with both high surface area and good bulk electrical conductivity is provided. A porous substrate having a thickness of 100 microns or more and comprising macropores throughout its thickness is prepared. At least one catalyst is deposited inside the porous substrate. Subsequently, chemical vapor deposition is used to uniformly deposit a nanomaterial in the macropores throughout the thickness of the porous substrate. Applications include electrical energy storage, such as batteries and capacitors, and hydrogen storage.

  4. Substrate mediated enzyme prodrug therapy

    DEFF Research Database (Denmark)

    Fejerskov, Betina; Jarlstad Olesen, Morten T; Zelikin, Alexander N

    2017-01-01

    Substrate mediated enzyme prodrug therapy (SMEPT) is a biomedical platform developed to perform a localized synthesis of drugs mediated by implantable biomaterials. This approach combines the benefits and at the same time offers to overcome the drawbacks for traditional pill-based drug administra......Substrate mediated enzyme prodrug therapy (SMEPT) is a biomedical platform developed to perform a localized synthesis of drugs mediated by implantable biomaterials. This approach combines the benefits and at the same time offers to overcome the drawbacks for traditional pill-based drug...

  5. Identification and characterisation of a novel acylpeptide hydrolase from Sulfolobus solfataricus: structural and functional insights.

    Directory of Open Access Journals (Sweden)

    Marta Gogliettino

    Full Text Available A novel acylpeptide hydrolase, named APEH-3(Ss, was isolated from the hypertermophilic archaeon Sulfolobus solfataricus. APEH is a member of the prolyl oligopeptidase family which catalyzes the removal of acetylated amino acid residues from the N terminus of oligopeptides. The purified enzyme shows a homotrimeric structure, unique among the associate partners of the APEH cluster and, in contrast to the archaeal APEHs which show both exo/endo peptidase activities, it appears to be a "true" aminopeptidase as exemplified by its mammalian counterparts, with which it shares a similar substrate specificity. Furthermore, a comparative study on the regulation of apeh gene expression, revealed a significant but divergent alteration in the expression pattern of apeh-3(Ss and apeh(Ss (the gene encoding the previously identified APEH(Ss from S. solfataricus, which is induced in response to various stressful growth conditions. Hence, both APEH enzymes can be defined as stress-regulated proteins which play a complementary role in enabling the survival of S. solfataricus cells under different conditions. These results provide new structural and functional insights into S. solfataricus APEH, offering a possible explanation for the multiplicity of this enzyme in Archaea.

  6. Andrographolide inhibits hypoxia-induced HIF-1α-driven endothelin 1 secretion by activating Nrf2/HO-1 and promoting the expression of prolyl hydroxylases 2/3 in human endothelial cells.

    Science.gov (United States)

    Lin, Hung-Chih; Su, Shih-Li; Lu, Chia-Yang; Lin, Ai-Hsuan; Lin, Wan-Chun; Liu, Chin-San; Yang, Ya-Chen; Wang, Hsiu-Miao; Lii, Chong-Kuei; Chen, Haw-Wen

    2017-03-01

    Andrographolide, the main bioactive component of the medicinal plant Andrographis paniculata, has been shown to possess potent anti-inflammatory activity. Endothelin 1 (ET-1), a potent vasoconstrictor peptide produced by vascular endothelial cells, displays proinflammatory property. Hypoxia-inducible factor 1α (HIF-1α), the regulatory member of the transcription factor heterodimer HIF-1α/β, is one of the most important molecules that responds to hypoxia. Changes in cellular HIF-1α protein level are the result of altered gene transcription and protein stability, with the latter being dependent on prolyl hydroxylases (PHDs). In this study, inhibition of pro-inflammatory ET-1 expression and changes of HIF-1α gene transcription and protein stability under hypoxia by andrographolide in EA.hy926 endothelial-like cells were investigated. Hypoxic conditions were created using the hypoxia-mimetic agent CoCl 2. We found that hypoxia stimulated the production of reactive oxygen species (ROS), the expression of HIF-1α mRNA and protein, and the expression and secretion of ET-1. These effects, however, were attenuated by co-exposure to andrographolide, bilirubin, and RuCO. Silencing Nrf2 and heme oxygenase 1 (HO-1) reversed the inhibitory effects of andrographolide on hypxoia-induced HIF-1α mRNA and protein expression. Moreover, andrographolide increased the expression of prolyl hydroxylases (PHD) 2/3, which hydroxylate HIF-1α and promotes HIF-1α proteasome degradation, with an increase in HIF-1α hydroxylation was noted under hypoxia. Inhibition of p38 MAPK abrogated the hypoxia-induced increases in HIF-1α mRNA and protein expression as well as ET-1 mRNA expression and secretion. Taken together, these results suggest that andrographolide suppresses hypoxia-induced pro-inflammatory ET-1 expression by activating Nrf2/HO-1, inhibiting p38 MAPK signaling, and promoting PHD2/3 expression. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 918-930, 2017. © 2016 Wiley

  7. Phonon scattering in graphene over substrate steps

    DEFF Research Database (Denmark)

    Sevincli, Haldun; Brandbyge, Mads

    2014-01-01

    We calculate the effect on phonon transport of substrate-induced bends in graphene. We consider bending induced by an abrupt kink in the substrate, and provide results for different step-heights and substrate interaction strengths. We find that individual substrate steps reduce thermal conductance...

  8. Neurobiological Substrates of Tourette's Disorder

    NARCIS (Netherlands)

    Leckman, James F.; Bloch, Michael H.; Smith, Megan E.; Larabi, Daouia; Hampson, Michelle

    Objective: This article reviews the available scientific literature concerning the neurobiological substrates of Tourette's disorder (TD). Methods: The electronic databases of PubMed, ScienceDirect, and PsycINFO were searched for relevant studies using relevant search terms. Results:

  9. Sensor Technologies on Flexible Substrates

    Science.gov (United States)

    Koehne, Jessica

    2016-01-01

    NASA Ames has developed sensor technologies on flexible substrates integrated into textiles for personalized environment monitoring and human performance evaluation. Current technologies include chemical sensing for gas leak and event monitoring and biological sensors for human health and performance monitoring. Targeted integration include next generation EVA suits and flexible habitats.

  10. Imparting Icephobicity with Substrate Flexibility

    Science.gov (United States)

    Schutzius, Thomas; Vasileiou, Thomas; Poulikakos, Dimos

    2017-11-01

    Ice accumulation poses serious safety and performance issues for modern infrastructure. Rationally designed superhydrophobic surfaces have demonstrated potential as a passive means to mitigate ice accretion; however, further studies on solutions that reduce impalement and contact time for impacting supercooled droplets are urgently needed. Here we demonstrate the collaborative effect of substrate flexibility and surface texture on enhancing icephobicity and repelling viscous droplets. We first investigate the influence of increased viscosity on impalement resistance and droplet-substrate contact time. Then we examine the effect of droplet partial solidification on recoil by impacting supercooled water droplets onto surfaces containing ice nucleation promoters. We demonstrate a passive method for shedding partially solidified droplets that does not rely on the classic recoil mechanism. Using an energy-based model, we identify a previously unexplored mechanism whereby the substrate oscillation governs the rebound process by efficiently absorbing the droplet kinetic energy and rectifying it back, allowing for droplet recoil. This mechanism applies for a range of droplet viscosities and ice slurries, which do not rebound from rigid superhydrophobic substrates. Partial support of the Swiss National Science Foundation under Grant No. 162565 and the European Research Council under Advanced Grant No. 669908 (INTICE) is acknowledged.

  11. PREFACE: Cell-substrate interactions Cell-substrate interactions

    Science.gov (United States)

    Gardel, Margaret; Schwarz, Ulrich

    2010-05-01

    One of the most striking achievements of evolution is the ability to build cellular systems that are both robust and dynamic. Taken by themselves, both properties are obvious requirements: robustness reflects the fact that cells are there to survive, and dynamics is required to adapt to changing environments. However, it is by no means trivial to understand how these two requirements can be implemented simultaneously in a physical system. The long and difficult quest to build adaptive materials is testimony to the inherent difficulty of this goal. Here materials science can learn a lot from nature, because cellular systems show that robustness and dynamics can be achieved in a synergetic fashion. For example, the capabilities of tissues to repair and regenerate are still unsurpassed in the world of synthetic materials. One of the most important aspects of the way biological cells adapt to their environment is their adhesive interaction with the substrate. Numerous aspects of the physiology of metazoan cells, including survival, proliferation, differentiation and migration, require the formation of adhesions to the cell substrate, typically an extracellular matrix protein. Adhesions guide these diverse processes both by mediating force transmission from the cell to the substrate and by controlling biochemical signaling pathways. While the study of cell-substrate adhesions is a mature field in cell biology, a quantitative biophysical understanding of how the interactions of the individual molecular components give rise to the rich dynamics and mechanical behaviors observed for cell-substrate adhesions has started to emerge only over the last decade or so. The recent growth of research activities on cell-substrate interactions was strongly driven by the introduction of new physical techniques for surface engineering into traditional cell biological work with cell culture. For example, microcontact printing of adhesive patterns was used to show that cell fate depends

  12. Stimulating retinal blood vessel protection with hypoxia-inducible factor stabilization: identification of novel small-molecule hydrazones to inhibit hypoxia-inducible factor prolyl hydroxylase (an American Ophthalmological Society thesis).

    Science.gov (United States)

    Sears, Jonathan E; Hoppe, George

    2013-09-01

    To discover novel small molecules that inhibit hypoxia-inducible factor (HIF) prolyl hydroxylase (PHD), a key enzyme that regulates the posttranslational stability and hence activity of HIF. NIH3T3 cell line stably transfected with firefly luciferase under a HIF-1-inducible promoter was used to screen a Chembridge library of 34,000 small molecules of molecular weight 250 to 550 Da. Positive hits were considered at 4.5-fold higher luminescence than control. Selected compounds were validated in vitro. The most effective dose was then used to treat mice expressing firefly luciferase fused to the oxygen-dependent degradation domain (lucODD) in order to determine the location of the receptor for systemic treatment with small-molecule HIF PHD inhibitors. Twenty-three novel small molecules were discovered, the majority of which were hydrazones and hydrazines. Of the 23 compounds, each had different selectivity for expression of erythropoietin or vascular endothelial growth factor, two angiogenic, HIF-regulated gene products. In addition, each showed different selectivity for hepatocytes or kidney, or both or neither, when injected intraperitoneally in an in vivo reporter gene assay. The discovery of multiple small molecules that inhibit HIF PHD identifies new reagents to develop strategies to prevent the degradation of HIF by its selective PHD. These molecules are novel hypoxia mimetics that may provide new strategies to protect retinovasculature from hyperoxia.

  13. Click Chemistry-based Discovery of [3-Hydroxy-5-(1H-1,2,3-triazol-4-yl)picolinoyl]glycines as Orally Active Hypoxia Inducing Factor Prolyl Hydroxylase Inhibitors with Favorable Safety Profiles for the Treatment of Anemia.

    Science.gov (United States)

    Wu, Yue; Jiang, Zhensheng; Li, Zhihong; Gu, Jing; You, Qi-Dong; Zhang, Xiaojin

    2018-06-01

    As a gene associated with anemia, the erythropoiesis gene is physiologically expressed under hypoxia regulated by hypoxia-inducing factor-α (HIF-α). Thus, stabilizing HIF-α is a potent strategy to stimulate the expression and secretion of erythropoiesis. In this study we applied click chemistry to the discovery of HIF prolyl hydroxylase 2 (HIF-PHD2) inhibitors for the first time and a series of triazole compounds showed preferable inhibitory activity in fluorescence polarization assay. Of particular note was the orally active HIF-PHD inhibitor 15i (IC50 = 62.23 nM), which was almost ten times more active than the phase III drug FG-4592 (IC50 = 591.4 nM). Furthermore, it can upregulate the hemoglobin of cisplatin induced anemia mice (120 g/L) to normal levels (160 g/L) with no apparent toxicity observed in vivo. These results confirm that triazole compound 15i is a promising candidate for the treatment of renal anemia.

  14. Verfahren zum Herstellen einer Beschichtung eines Substrats

    OpenAIRE

    Wilke, Martin; Töpper, Michael

    2013-01-01

    The method involves applying coating material (7) on surface (2) of recess (3) formed in substrate (1). A liquid auxiliary agent (6) is applied on substrate surface, such that recess is filled with auxiliary agent. The coating material is subsequently applied to auxiliary agent on substrate. A coating material portion in auxiliary agent is transported by coating material diffusion. The agent is subsequently separated from coating material, such that coating material on substrate surface is le...

  15. SUPPLEMENTARY INFORMATION Indicators for suicide substrate ...

    Indian Academy of Sciences (India)

    Jatinder

    The usual trend is to apply QSSA to a system with high substrate concentration. But, QSSA, i.e., steadiness in intermediate concentration, may even be achieved at high and even comparable enzyme-substrate ratio. Whether a system will attain a steady state depends not only on the high substrate concentration, but also on ...

  16. Method for coating substrates and mask holder

    NARCIS (Netherlands)

    Bijkerk, Frederik; Yakshin, Andrey; Louis, Eric; Kessels, M.J.H.; Maas, Edward Lambertus Gerardus; Bruineman, Caspar

    2004-01-01

    When coating substrates it is frequently desired that the layer thickness should be a certain function of the position on the substrate to be coated. To control the layer thickness a mask is conventionally arranged between the coating particle source and the substrate. This leads to undesirable

  17. Superhydrophobicity enhancement through substrate flexibility

    Science.gov (United States)

    Vasileiou, Thomas; Gerber, Julia; Prautzsch, Jana; Schutzius, Thomas; Poulikakos, Dimos

    2017-11-01

    Inspired by manifestations in nature, micro/nanoengineering superhydrophobic surfaces has been the focus of much work. Generally, hydrophobicity is increased through the combined effects of surface texturing and chemistry; being durable, rigid substrate materials are the norm. However, many natural and technical materials are flexible, and the resulting effect on hydrophobicity has been largely unexplored. Here, we show that the rational tuning of flexibility can work collaboratively with the surface micro/nanotexture to enhance liquid repellency performance, defined by impalement and breakup resistance, contact time reduction, and restitution coefficient increase. Reduction in substrate stiffness and areal density imparts immediate acceleration and intrinsic responsiveness to impacting droplets, mitigating the collision and lowering the impalement probability by 60 % without the need for active actuation. We demonstrate the above discoveries with materials ranging from thin steel or polymer sheets to butterfly wings. Partial support of the Swiss National Science Foundation under Grant 162565 and the European Research Council under Advanced Grant 669908 (INTICE) is acknowledged.

  18. Quartz substrate infrared photonic crystal

    Science.gov (United States)

    Ghadiri, Khosrow; Rejeb, Jalel; Vitchev, Vladimir N.

    2003-01-01

    This paper presents the fabrication of a planar photonic crystal (p2c) made of a square array of dielectric rods embedded in air, operating in the infrared spectrum. A quartz substrate is employed instead of the commonly used silicon or column III-V substrate. Our square structure has a normalized cylinder radius-to-pitch ratio of r/a = 0.248 and dielectric material contrast ɛr of 4.5. We choose a Z-cut synthetic quartz for its cut (geometry), and etching properties. Then a particular Z-axis etching process is employed in order to ensure the sharp-edged verticality of the rods and fast etching speed. We also present the computer simulations that allowed the establishment of the photonic band gaps (PBG) of our photonic crystal, as well as the actual measurements. An experimental measurement have been carried out and compared with different simulations. It was found that experimental results are in good agreement with different simulation results. Finally, a frequency selective device for optical communication based on the introduction of impurity sites in the photonic crystal is presented. With our proposed structure Optical System on a Chip (OsoC) with micro-cavity based active devices such as lasers, diodes, modulators, couplers, frequency selective emitters, add-drop filters, detectors, mux/demuxes and polarizers connected by passive waveguide links can be realized.

  19. Substrate mediated enzyme prodrug therapy.

    Directory of Open Access Journals (Sweden)

    Betina Fejerskov

    Full Text Available In this report, we detail Substrate Mediated Enzyme Prodrug Therapy (SMEPT as a novel approach in drug delivery which relies on enzyme-functionalized cell culture substrates to achieve a localized conversion of benign prodrug(s into active therapeutics with subsequent delivery to adhering cells or adjacent tissues. For proof-of-concept SMEPT, we use surface adhered micro-structured physical hydrogels based on poly(vinyl alcohol, β-glucuronidase enzyme and glucuronide prodrugs. We demonstrate enzymatic activity mediated by the assembled hydrogel samples and illustrate arms of control over rate of release of model fluorescent cargo. SMEPT was not impaired by adhering cells and afforded facile time - and dose - dependent uptake of the in situ generated fluorescent cargo by hepatic cells, HepG2. With the use of a glucuronide derivative of an anticancer drug, SN-38, SMEPT afforded a decrease in cell viability to a level similar to that achieved using parent drug. Finally, dose response was achieved using SMEPT and administration of judiciously chosen concentration of SN-38 glucuronide prodrug thus revealing external control over drug delivery using drug eluting surface. We believe that this highly adaptable concept will find use in diverse biomedical applications, specifically surface mediated drug delivery and tissue engineering.

  20. Automated cassette-to-cassette substrate handling system

    Science.gov (United States)

    Kraus, Joseph Arthur; Boyer, Jeremy James; Mack, Joseph; DeChellis, Michael; Koo, Michael

    2014-03-18

    An automated cassette-to-cassette substrate handling system includes a cassette storage module for storing a plurality of substrates in cassettes before and after processing. A substrate carrier storage module stores a plurality of substrate carriers. A substrate carrier loading/unloading module loads substrates from the cassette storage module onto the plurality of substrate carriers and unloads substrates from the plurality of substrate carriers to the cassette storage module. A transport mechanism transports the plurality of substrates between the cassette storage module and the plurality of substrate carriers and transports the plurality of substrate carriers between the substrate carrier loading/unloading module and a processing chamber. A vision system recognizes recesses in the plurality of substrate carriers corresponding to empty substrate positions in the substrate carrier. A processor receives data from the vision system and instructs the transport mechanism to transport substrates to positions on the substrate carrier in response to the received data.

  1. Method For Producing Mechanically Flexible Silicon Substrate

    KAUST Repository

    Hussain, Muhammad Mustafa

    2014-08-28

    A method for making a mechanically flexible silicon substrate is disclosed. In one embodiment, the method includes providing a silicon substrate. The method further includes forming a first etch stop layer in the silicon substrate and forming a second etch stop layer in the silicon substrate. The method also includes forming one or more trenches over the first etch stop layer and the second etch stop layer. The method further includes removing the silicon substrate between the first etch stop layer and the second etch stop layer.

  2. Phonon scattering in graphene over substrate steps

    International Nuclear Information System (INIS)

    Sevinçli, H.; Brandbyge, M.

    2014-01-01

    We calculate the effect on phonon transport of substrate-induced bends in graphene. We consider bending induced by an abrupt kink in the substrate, and provide results for different step-heights and substrate interaction strengths. We find that individual substrate steps reduce thermal conductance in the range between 5% and 47%. We also consider the transmission across linear kinks formed by adsorption of atomic hydrogen at the bends and find that individual kinks suppress thermal conduction substantially, especially at high temperatures. Our analysis show that substrate irregularities can be detrimental for thermal conduction even for small step heights.

  3. Method For Producing Mechanically Flexible Silicon Substrate

    KAUST Repository

    Hussain, Muhammad Mustafa; Rojas, Jhonathan Prieto

    2014-01-01

    A method for making a mechanically flexible silicon substrate is disclosed. In one embodiment, the method includes providing a silicon substrate. The method further includes forming a first etch stop layer in the silicon substrate and forming a second etch stop layer in the silicon substrate. The method also includes forming one or more trenches over the first etch stop layer and the second etch stop layer. The method further includes removing the silicon substrate between the first etch stop layer and the second etch stop layer.

  4. Non-permeable substrate carrier for electroplating

    Science.gov (United States)

    Abas, Emmanuel Chua; Chen, Chen-An; Ma, Diana Xiaobing; Ganti, Kalyana Bhargava

    2012-11-27

    One embodiment relates to a substrate carrier for use in electroplating a plurality of substrates. The substrate carrier comprises a non-conductive carrier body on which the substrates are to be held. Electrically-conductive lines are embedded within the carrier body, and a plurality of contact clips are coupled to the electrically-conductive lines embedded within the carrier body. The contact clips hold the substrates in place and electrically couple the substrates to the electrically-conductive lines. The non-conductive carrier body is continuous so as to be impermeable to flow of electroplating solution through the non-conductive carrier body. Other embodiments, aspects and features are also disclosed.

  5. Identification of oxidized protein hydrolase as a potential prodrug target in prostate cancer

    International Nuclear Information System (INIS)

    McGoldrick, Christopher A; Jiang, Yu-Lin; Paromov, Victor; Brannon, Marianne; Krishnan, Koyamangalath; Stone, William L

    2014-01-01

    Esterases are often overexpressed in cancer cells and can have chiral specificities different from that of the corresponding normal tissues. For this reason, ester prodrugs could be a promising approach in chemotherapy. In this study, we focused on the identification and characterization of differentially expressed esterases between non-tumorigenic and tumorigenic prostate epithelial cells. Cellular lysates from LNCaP, DU 145, and PC3 prostate cancer cell lines, tumorigenic RWPE-2 prostate epithelial cells, and non-tumorigenic RWPE-1 prostate epithelial cells were separated by native polyacrylamide gel electrophoresis (n-PAGE) and the esterase activity bands visualized using α-naphthyl acetate or α-naphthyl-N-acetylalaninate (ANAA) chiral esters and Fast Blue RR salt. The esterases were identified using nanospray LC/MS-MS tandem mass spectrometry and confirmed by Western blotting, native electroblotting, inhibition assays, and activity towards a known specific substrate. The serine protease/esterase oxidized protein hydrolase (OPH) was overexpressed in COS-7 cells to verify our results. The major esterase observed with the ANAA substrates within the n-PAGE activity bands was identified as OPH. OPH (EC 3.4.19.1) is a serine protease/esterase and a member of the prolyl oligopeptidase family. We found that LNCaP lysates contained approximately 40% more OPH compared to RWPE-1 lysates. RWPE-2, DU145 and PC3 cell lysates had similar levels of OPH activity. OPH within all of the cell lysates tested had a chiral preference for the S-isomer of ANAA. LNCaP cells were stained more intensely with ANAA substrates than RWPE-1 cells and COS-7 cells overexpressing OPH were found to have a higher activity towards the ANAA and AcApNA than parent COS-7 cells. These data suggest that prodrug derivatives of ANAA and AcApNA could have potential as chemotherapeutic agents for the treatment of prostate cancer tumors that overexpress OPH

  6. Optically controlled electrophoresis with a photoconductive substrate

    Science.gov (United States)

    Inami, Wataru; Nagashima, Taiki; Kawata, Yoshimasa

    2018-05-01

    A photoconductive substrate is used to perform electrophoresis. Light-induced micro-particle flow manipulation is demonstrated without using a fabricated flow channel. The path along which the particles were moved was formed by an illuminated light pattern on the substrate. Because the substrate conductivity and electric field distribution can be modified by light illumination, the forces acting on the particles can be controlled. This technique has potential applications as a high functionality analytical device.

  7. Substrate optimization for integrated circuit antennas

    OpenAIRE

    Alexopoulos, N. G.; Katehi, P. B.; Rutledge, D. B.

    1982-01-01

    Imaging systems in microwaves, millimeter and submillimeter wave applications employ printed circuit antenna elements. The effect of substrate properties is analyzed in this paper by both reciprocity theorem as well as integral equation approach for infinitesimally short as well as finite length dipole and slot elements. Radiation efficiency and substrate surface wave guidance is studied for practical substrate materials as GaAs, Silicon, Quartz and Duroid.

  8. Adhesion of rhodium films on metallic substrates

    International Nuclear Information System (INIS)

    Marot, L.; Covarel, G.; Tuilier, M.-H.; Steiner, R.; Oelhafen, P.

    2008-01-01

    Rhodium coated metallic films were prepared by magnetron sputtering on metallic substrates. All films were elaborated in same conditions on copper, molybdenum and stainless steel. Adhesion strength tests were carried out by scratch test. The results reveal that the adhesion strength between the film and the substrate is influenced by the hardness of the substrate. Increase of deposition temperature improves the adhesion of the coating. In addition, pre-treatment of substrates by a filtered cathodic vacuum arc and the layer thickness have has some effects on the final adhesion strength

  9. Adhesion of rhodium films on metallic substrates

    Energy Technology Data Exchange (ETDEWEB)

    Marot, L. [Department of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland)], E-mail: laurent.marot@unibas.ch; Covarel, G.; Tuilier, M.-H. [Laboratoire Mecanique, Materiaux et Procedes de Fabrication, Pole STIC-SPI-Math 61 rue Albert Camus, Universite de Haute-Alsace, F-68093 - Mulhouse Cedex (France); Steiner, R.; Oelhafen, P. [Department of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel (Switzerland)

    2008-09-01

    Rhodium coated metallic films were prepared by magnetron sputtering on metallic substrates. All films were elaborated in same conditions on copper, molybdenum and stainless steel. Adhesion strength tests were carried out by scratch test. The results reveal that the adhesion strength between the film and the substrate is influenced by the hardness of the substrate. Increase of deposition temperature improves the adhesion of the coating. In addition, pre-treatment of substrates by a filtered cathodic vacuum arc and the layer thickness have has some effects on the final adhesion strength.

  10. Nanowires and nanoneedles nucleation on vicinal substrate

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Xu, E-mail: zhangxubetter@gmail.com [Henan Key Laboratory of Laser and Opto-electric Information Technology, School of Information Engineering, Zhengzhou University, Zhengzhou 450052 (China); Xie, Dan; Huang, Genling [Zhengzhou Railway Vocational and Technical College, Zhengzhou 450052 (China); Sun, Xiao-Hong [Henan Key Laboratory of Laser and Opto-electric Information Technology, School of Information Engineering, Zhengzhou University, Zhengzhou 450052 (China)

    2015-01-01

    An analytic stress-driven nucleation model of nanowires (NWs) and nanoneedles (NNs) growing on a mismatched vicinal substrate is proposed. It is demonstrated that the formation enthalpy of NWs and NNs is a function of three independent variables, the base radius, aspect ratio and miscut angle of the vicinal surface. Theoretical analysis shows that the minimum nucleation barrier of an island decreases with increment of substrate misorientation, which means the nucleation of islands on a vicinal substrate is more favorable than that on a flat substrate.

  11. Molecular characterization and mRNA expression of two key enzymes of hypoxia-sensing pathways in eastern oysters Crassostrea virginica (Gmelin): Hypoxia-inducible factor α (HIF-α) and HIF-prolyl hydroxylase (PHD)

    Science.gov (United States)

    Piontkivska, Helen; Chung, J. Sook; Ivanina, Anna V.; Sokolov, Eugene P.; Techa, Sirinart; Sokolova, Inna M.

    2010-01-01

    Oxygen homeostasis is crucial for development, survival and normal function of all metazoans. A family of transcription factors called hypoxia-inducible factors (HIF) is critical in mediating the adaptive responses to reduced oxygen availability. The HIF transcription factor consists of a constitutively expressed β subunit and an oxygen-dependent α subunit; the abundance of the latter determines the activity of HIF and is regulated by a family of O2- and Fe2+-dependent enzymes prolyl hydroxylases (PHDs). Currently very little is known about the function of this important pathway and the molecular structure of its key players in hypoxia-tolerant intertidal mollusks including oysters, which are among the animal champions of anoxic and hypoxic tolerance and thus can serve as excellent models to study the role of HIF cascade in adaptations to oxygen deficiency. We have isolated transcripts of two key components of the oxygen sensing pathway - the oxygen-regulated HIF-α subunit and PHD - from an intertidal mollusk, the eastern oyster Crassostrea virginica, and determined the transcriptional responses of these two genes to anoxia, hypoxia and cadmium (Cd) stress. HIF-α and PHD homologs from eastern oysters C. virginica show significant sequence similarity and share key functional domains with the earlier described isoforms from vertebrates and invertebrates. Phylogenetic analysis shows that genetic diversification of HIF and PHD isoforms occurred within the vertebrate lineage indicating functional diversification and specialization of the oxygen-sensing pathways in this group, which parallels situation observed for many other important genes. HIF-α and PHD homologs are broadly expressed at the mRNA level in different oyster tissues and show transcriptional responses to prolonged hypoxia in the gills consistent with their putative role in oxygen sensing and the adaptive response to hypoxia. Similarity in amino acid sequence, domain structure and transcriptional

  12. Organism-substrate relationships in lowland streams

    NARCIS (Netherlands)

    Tolkamp, H.H.

    1980-01-01

    A field and laboratory study on the microdistribution of bottom dwelling macroinvertebrates to investigate the role of the stream substrate In the development and preservation of the macroinvertebrate communities in natural, undisturbed lowland streams is described. Field data on bottom substrates

  13. Cellulose Nanofiber Composite Substrates for Flexible Electronics

    Science.gov (United States)

    Ronald Sabo; Jung-Hun Seo; Zhenqiang Ma

    2012-01-01

    Flexible electronics have a large number of potential applications including malleable displays and wearable computers. The current research into high-speed, flexible electronic substrates employs the use of plastics for the flexible substrate, but these plastics typically have drawbacks, such as high thermal expansion coefficients. Transparent films made from...

  14. Direct transfer of graphene onto flexible substrates.

    Science.gov (United States)

    Martins, Luiz G P; Song, Yi; Zeng, Tingying; Dresselhaus, Mildred S; Kong, Jing; Araujo, Paulo T

    2013-10-29

    In this paper we explore the direct transfer via lamination of chemical vapor deposition graphene onto different flexible substrates. The transfer method investigated here is fast, simple, and does not require an intermediate transfer membrane, such as polymethylmethacrylate, which needs to be removed afterward. Various substrates of general interest in research and industry were studied in this work, including polytetrafluoroethylene filter membranes, PVC, cellulose nitrate/cellulose acetate filter membranes, polycarbonate, paraffin, polyethylene terephthalate, paper, and cloth. By comparing the properties of these substrates, two critical factors to ensure a successful transfer on bare substrates were identified: the substrate's hydrophobicity and good contact between the substrate and graphene. For substrates that do not satisfy those requirements, polymethylmethacrylate can be used as a surface modifier or glue to ensure successful transfer. Our results can be applied to facilitate current processes and open up directions for applications of chemical vapor deposition graphene on flexible substrates. A broad range of applications can be envisioned, including fabrication of graphene devices for opto/organic electronics, graphene membranes for gas/liquid separation, and ubiquitous electronics with graphene.

  15. Metal oxide nanorod arrays on monolithic substrates

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Pu-Xian; Guo, Yanbing; Ren, Zheng

    2018-01-02

    A metal oxide nanorod array structure according to embodiments disclosed herein includes a monolithic substrate having a surface and multiple channels, an interface layer bonded to the surface of the substrate, and a metal oxide nanorod array coupled to the substrate surface via the interface layer. The metal oxide can include ceria, zinc oxide, tin oxide, alumina, zirconia, cobalt oxide, and gallium oxide. The substrate can include a glass substrate, a plastic substrate, a silicon substrate, a ceramic monolith, and a stainless steel monolith. The ceramic can include cordierite, alumina, tin oxide, and titania. The nanorod array structure can include a perovskite shell, such as a lanthanum-based transition metal oxide, or a metal oxide shell, such as ceria, zinc oxide, tin oxide, alumina, zirconia, cobalt oxide, and gallium oxide, or a coating of metal particles, such as platinum, gold, palladium, rhodium, and ruthenium, over each metal oxide nanorod. Structures can be bonded to the surface of a substrate and resist erosion if exposed to high velocity flow rates.

  16. Microbial growth and substrate utilization kinetics | Okpokwasili ...

    African Journals Online (AJOL)

    Microbial growth on and utilization of environmental contaminants as substrates have been studied by many researchers. Most times, substrate utilization results in removal of chemical contaminant, increase in microbial biomass and subsequent biodegradation of the contaminant. These are all aimed at detoxification of the ...

  17. AFM plough YBCO micro bridges: substrate effects

    CSIR Research Space (South Africa)

    Elkaseh, A

    2009-04-01

    Full Text Available AFM nanolithography was used as a novel cutting technique to define micro-size YBCO superconducting constrictions. Researchers studied the substrate effects on MgO and STO substrates and showed that the observed Shapiro steps from the bridges on STO...

  18. Substrate tolerant direct block copolymer nanolithography

    DEFF Research Database (Denmark)

    Li, Tao; Wang, Zhongli; Schulte, Lars

    2016-01-01

    Block copolymer (BC) self-assembly constitutes a powerful platform for nanolithography. However, there is a need for a general approach to BC lithography that critically considers all the steps from substrate preparation to the final pattern transfer. We present a procedure that significantly sim...... plasma treatment enables formation of the oxidized PDMS hard mask, PS block removal and polymer or graphene substrate patterning....

  19. A signal-substrate match in the substrate-borne component of a multimodal courtship display

    Directory of Open Access Journals (Sweden)

    Damian O. ELIAS, Andrew C. MASON, Eileen A. HEBETS

    2010-06-01

    Full Text Available The environment can impose strong limitations on the efficacy of signal transmission. In particular, for vibratory communication, the signaling environment is often extremely heterogeneous at very small scales. Nevertheless, natural selection is expected to select for signals well-suited to effective transmission. Here, we test for substrate-dependent signal efficacy in the wolf spider Schizocosa stridulans Stratton 1991. We first explore the transmission characteristics of this important signaling modality by playing recorded substrate-borne signals through three different substrates (leaf litter, pine litter, and red clay and measuring the propagated signal. We found that the substrate-borne signal of S. stridulans attenuates the least on leaf litter, the substrate upon which the species is naturally found. Next, by assessing mating success with artificially muted and non-muted males across different signaling substrates (leaf litter, pine litter, and sand, we explored the relationship between substrate-borne signaling and substrate for mating success. We found that muted males were unsuccessful in obtaining copulations regardless of substrate, while mating success was dependent on the signaling substrate for non-muted males. For non-muted males, more males copulated on leaf litter than any other substrate. Taken together, these results confirm the importance of substrate-borne signaling in S. stridulans and suggest a match between signal properties and signal efficacy – leaf litter transmits the signal most effectively and males are most successful in obtaining copulations on leaf litter [Current Zoology 56 (3: 370–378, 2010].

  20. Alternative Substrate Metabolism in Yarrowia lipolytica

    Directory of Open Access Journals (Sweden)

    Michael Spagnuolo

    2018-05-01

    Full Text Available Recent advances in genetic engineering capabilities have enabled the development of oleochemical producing strains of Yarrowia lipolytica. Much of the metabolic engineering effort has focused on pathway engineering of the product using glucose as the feedstock; however, alternative substrates, including various other hexose and pentose sugars, glycerol, lipids, acetate, and less-refined carbon feedstocks, have not received the same attention. In this review, we discuss recent work leading to better utilization of alternative substrates. This review aims to provide a comprehensive understanding of the current state of knowledge for alternative substrate utilization, suggest potential pathways identified through homology in the absence of prior characterization, discuss recent work that either identifies, endogenous or cryptic metabolism, and describe metabolic engineering to improve alternative substrate utilization. Finally, we describe the critical questions and challenges that remain for engineering Y. lipolytica for better alternative substrate utilization.

  1. Modelling substrate specificity and enantioselectivity for lipases and esterases by substrate-imprinted docking

    Directory of Open Access Journals (Sweden)

    Tyagi Sadhna

    2009-06-01

    Full Text Available Abstract Background Previously, ways to adapt docking programs that were developed for modelling inhibitor-receptor interaction have been explored. Two main issues were discussed. First, when trying to model catalysis a reaction intermediate of the substrate is expected to provide more valid information than the ground state of the substrate. Second, the incorporation of protein flexibility is essential for reliable predictions. Results Here we present a predictive and robust method to model substrate specificity and enantioselectivity of lipases and esterases that uses reaction intermediates and incorporates protein flexibility. Substrate-imprinted docking starts with covalent docking of reaction intermediates, followed by geometry optimisation of the resulting enzyme-substrate complex. After a second round of docking the same substrate into the geometry-optimised structures, productive poses are identified by geometric filter criteria and ranked by their docking scores. Substrate-imprinted docking was applied in order to model (i enantioselectivity of Candida antarctica lipase B and a W104A mutant, (ii enantioselectivity and substrate specificity of Candida rugosa lipase and Burkholderia cepacia lipase, and (iii substrate specificity of an acetyl- and a butyrylcholine esterase toward the substrates acetyl- and butyrylcholine. Conclusion The experimentally observed differences in selectivity and specificity of the enzymes were reproduced with an accuracy of 81%. The method was robust toward small differences in initial structures (different crystallisation conditions or a co-crystallised ligand, although large displacements of catalytic residues often resulted in substrate poses that did not pass the geometric filter criteria.

  2. Direct transfer of graphene onto flexible substrates

    Science.gov (United States)

    Martins, Luiz G. P.; Song, Yi; Zeng, Tingying; Dresselhaus, Mildred S.; Kong, Jing; Araujo, Paulo T.

    2013-01-01

    In this paper we explore the direct transfer via lamination of chemical vapor deposition graphene onto different flexible substrates. The transfer method investigated here is fast, simple, and does not require an intermediate transfer membrane, such as polymethylmethacrylate, which needs to be removed afterward. Various substrates of general interest in research and industry were studied in this work, including polytetrafluoroethylene filter membranes, PVC, cellulose nitrate/cellulose acetate filter membranes, polycarbonate, paraffin, polyethylene terephthalate, paper, and cloth. By comparing the properties of these substrates, two critical factors to ensure a successful transfer on bare substrates were identified: the substrate’s hydrophobicity and good contact between the substrate and graphene. For substrates that do not satisfy those requirements, polymethylmethacrylate can be used as a surface modifier or glue to ensure successful transfer. Our results can be applied to facilitate current processes and open up directions for applications of chemical vapor deposition graphene on flexible substrates. A broad range of applications can be envisioned, including fabrication of graphene devices for opto/organic electronics, graphene membranes for gas/liquid separation, and ubiquitous electronics with graphene. PMID:24127582

  3. Method of beryllium implantation in germanium substrate

    International Nuclear Information System (INIS)

    Kagawa, S.; Baba, Y.; Kaneda, T.; Shirai, T.

    1983-01-01

    A semiconductor device is disclosed, as well as a method for manufacturing it in which ions of beryllium are implanted into a germanium substrate to form a layer containing p-type impurity material. There after the substrate is heated at a temperature in the range of 400 0 C. to 700 0 C. to diffuse the beryllium ions into the substrate so that the concentration of beryllium at the surface of the impurity layer is in the order of 10 17 cm- 3 or more. In one embodiment, a p-type channel stopper is formed locally in a p-type germanium substrate and an n-type active layer is formed in a region surrounded by, and isolated from, the channel stopper region. In another embodiment, a relatively shallow p-type active layer is formed at one part of an n-type germanium substrate and p-type guard ring regions are formed surrounding, and partly overlapping said p-type active layer. In a further embodiment, a p-type island region is formed at one part of an n-type germanium substrate, and an n-type region is formed within said p-type region. In these embodiments, the p-type channel stopper region, p-type guard ring regions and the p-type island region are all formed by implanting ions of beryllium into the germanium substrate

  4. Dynamics of Preferential Substrate Recognition in HIV-1 Protease: Redefining the Substrate Envelope

    Science.gov (United States)

    Özen, Ayşegül; Haliloğlu, Türkan; Schiffer, Celia A.

    2011-01-01

    HIV-1 protease (PR) permits viral maturation by processing the Gag and Gag-Pro-Pol polyproteins. Though HIV-1 PR inhibitors (PIs) are used in combination antiviral therapy, the emergence of drug resistance has limited their efficacy. The rapid evolution of HIV-1 necessitates the consideration of drug resistance in novel drug-design strategies. Drug-resistant HIV-1 PR variants, while no longer efficiently inhibited, continue to efficiently hydrolyze the natural viral substrates. Though highly diverse in sequence, the HIV-1 PR substrates bind in a conserved three-dimensional shape we defined as the “substrate envelope”. We previously showed that resistance mutations arise where PIs protrude beyond the substrate envelope, as these regions are crucial for drug binding but not for substrate recognition. Here, we extend this model by considering the role of protein dynamics in the interaction of HIV-1 PR with its substrates. Seven molecular dynamics simulations of PR-substrate complexes were performed to estimate the conformational flexibility of substrates in their complexes. Interdependency of the substrate-protease interactions may compensate for the variations in cleavage-site sequences, and explain how a diverse set of sequences can be recognized as substrates by the same enzyme. This diversity may be essential for regulating sequential processing of substrates. We also define a dynamic substrate envelope as a more accurate representation of PR-substrate interactions. This dynamic substrate envelope, described by a probability distribution function, is a powerful tool for drug design efforts targeting ensembles of resistant HIV-1 PR variants with the aim of developing drugs that are less susceptible to resistance. PMID:21762811

  5. Targeting Biological Sensing with Commercial SERS Substrates

    Science.gov (United States)

    2012-09-01

    substrate substrate. Sl x 4 mm waf urement to re ossible contam substrates, fiv resented as an e being used f tigated, and nam eived in an ac cations...targeted nanotechnology-based inhalation co-delivery of anticancer drugs and siRNA," Journal of Drug Targeting, 900-914 (2011). [35] Jong , H.J., Na...J.H., Jin, B.S., Lee, W.K., Lee, W.H., Jung, H.J., Kim , S.C., Lim, S.H., Yu, Y.G., "Identification of Dinitrotoluene Selective Peptides by Phage

  6. Ion implantation methods for semiconductor substrates

    International Nuclear Information System (INIS)

    Matsushita, T.; Mamine, T.; Hayashi, H.; Nishiyama, K.

    1980-01-01

    A method of ion implantation for controlling the life time of minority carriers in a semiconductor substrate and hence to reduce the temperature dependency of the life time, comprises implanting iron ions into an N type semiconductor substrate with a dosage of 10 10 to 10 15 ions cm -2 , and then heat-treating the implanted substrate at 850 0 to 1250 0 C. The method is applicable to the production of diodes, transistors, Si controlled rectifiers and gate controlled switching devices. (author)

  7. Flexible and foldable paper-substrate thermoelectric generator (teg)

    KAUST Repository

    Rojas, Jhonathan Prieto

    2017-08-24

    Flexible and foldable paper-substrate thermoelectric generators (TEGs) and methods for making the paper-substrate TEGs are disclosed. A method includes depositing a plurality of thermocouples in series on a paper substrate to create a paper-substrate TEG, wherein the plurality of thermocouples is deposited between two contact points of the paper-substrate TEG. The method may also include setting the power density and maximum achievable temperature gradient of the paper-substrate TEG by folding the paper-substrate TEG. A paper-substrate TEG apparatus may include a paper substrate and a plurality of thermocouples deposited in series on the paper substrate between two contact points of the paper-substrate TEG, wherein the power density and maximum achievable temperature gradient of the paper-substrate TEG is set by folding the paper-substrate TEG.

  8. Substrate curvature gradient drives rapid droplet motion.

    Science.gov (United States)

    Lv, Cunjing; Chen, Chao; Chuang, Yin-Chuan; Tseng, Fan-Gang; Yin, Yajun; Grey, Francois; Zheng, Quanshui

    2014-07-11

    Making small liquid droplets move spontaneously on solid surfaces is a key challenge in lab-on-chip and heat exchanger technologies. Here, we report that a substrate curvature gradient can accelerate micro- and nanodroplets to high speeds on both hydrophilic and hydrophobic substrates. Experiments for microscale water droplets on tapered surfaces show a maximum speed of 0.42  m/s, 2 orders of magnitude higher than with a wettability gradient. We show that the total free energy and driving force exerted on a droplet are determined by the substrate curvature and substrate curvature gradient, respectively. Using molecular dynamics simulations, we predict nanoscale droplets moving spontaneously at over 100  m/s on tapered surfaces.

  9. Iron films deposited on porous alumina substrates

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, Yasuhiro, E-mail: yyasu@rs.kagu.tus.ac.jp; Tanabe, Kenichi; Nishida, Naoki [Tokyo University of Science (Japan); Kobayashi, Yoshio [The University of Electro-Communications (Japan)

    2016-12-15

    Iron films were deposited on porous alumina substrates using an arc plasma gun. The pore sizes (120 – 250 nm) of the substrates were controlled by changing the temperature during the anodic oxidation of aluminum plates. Iron atoms penetrated into pores with diameters of less than 160 nm, and were stabilized by forming γ-Fe, whereas α-Fe was produced as a flat plane covering the pores. For porous alumina substrates with pore sizes larger than 200 nm, the deposited iron films contained many defects and the resulting α-Fe had smaller hyperfine magnetic fields. In addition, only a very small amount of γ-Fe was obtained. It was demonstrated that the composition and structure of an iron film can be affected by the surface morphology of the porous alumina substrate on which the film is grown.

  10. Transformation kinetics of mixed polymeric substrates under ...

    African Journals Online (AJOL)

    bglucosidase and a-mannosidase were abundantly secreted in the growth medium. This research is the first report on mixed polymeric substrate biodegradation under sewer condition by A. niger, and could be considered as an open window on ...

  11. Palladium clusters deposited on the heterogeneous substrates

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Kun, E-mail: cqdxwk@126.com [College of Power Engineering, Chongqing University, Chongqing 400044 (China); Key Laboratory of Low-grade Energy Utilization Technologies and Systems, Ministry of Education of PRC, Chongqing 400044 (China); Liu, Juanfang, E-mail: juanfang@cqu.edu.cn [College of Power Engineering, Chongqing University, Chongqing 400044 (China); Key Laboratory of Low-grade Energy Utilization Technologies and Systems, Ministry of Education of PRC, Chongqing 400044 (China); Chen, Qinghua, E-mail: qhchen@cqu.edu.cn [College of Power Engineering, Chongqing University, Chongqing 400044 (China); Key Laboratory of Low-grade Energy Utilization Technologies and Systems, Ministry of Education of PRC, Chongqing 400044 (China)

    2016-07-15

    Graphical abstract: The site-exchange between the substrate and cluster atoms can result in the formation of the surface alloys and the reconstruction of the cluster structure before the collision system approaching the thermal equilibrium. The deposited cluster adjusted the atom arrangement as possibly as to match the substrate lattice arrangement from bottom to up. The structural reconstruction is accompanied by the system potential energy minimization. - Highlights: • The deposition process can divide explicitly into three stages: adsorption, collision, relaxation. • The local melt does not emerge inside the substrate during the deposition process. • Surface alloys are formed by the site-exchange between the cluster and substrate atoms. • The cluster reconstructs the atom arrangement following as the substrate lattice arrangement from bottom to up. • The structural reconstruction ability and scope depend on the cluster size and incident energy. - Abstract: To improve the performance of the Pd composite membrane prepared by the cold spraying technology, it is extremely essential to give insights into the deposition process of the cluster and the heterogeneous deposition of the big Pd cluster at the different incident velocities on the atomic level. The deposition behavior, morphologies, energetic and interfacial configuration were examined by the molecular dynamic simulation and characterized by the cluster flattening ratio, the substrate maximum local temperature, the atom-embedded layer number and the surface-alloy formation. According to the morphology evolution, three deposition stages and the corresponding structural and energy evolution were clearly identified. The cluster deformation and penetrating depth increased with the enhancement of the incident velocity, but the increase degree also depended on the substrate hardness. The interfacial interaction between the cluster and the substrate can be improved by the higher substrate local temperature

  12. Fermentative hydrogen production from agroindustrial lignocellulosic substrates

    Science.gov (United States)

    Reginatto, Valeria; Antônio, Regina Vasconcellos

    2015-01-01

    To achieve economically competitive biological hydrogen production, it is crucial to consider inexpensive materials such as lignocellulosic substrate residues derived from agroindustrial activities. It is possible to use (1) lignocellulosic materials without any type of pretreatment, (2) lignocellulosic materials after a pretreatment step, and (3) lignocellulosic materials hydrolysates originating from a pretreatment step followed by enzymatic hydrolysis. According to the current literature data on fermentative H2 production presented in this review, thermophilic conditions produce H2 in yields approximately 75% higher than those obtained in mesophilic conditions using untreated lignocellulosic substrates. The average H2 production from pretreated material is 3.17 ± 1.79 mmol of H2/g of substrate, which is approximately 50% higher compared with the average yield achieved using untreated materials (2.17 ± 1.84 mmol of H2/g of substrate). Biological pretreatment affords the highest average yield 4.54 ± 1.78 mmol of H2/g of substrate compared with the acid and basic pretreatment - average yields of 2.94 ± 1.85 and 2.41 ± 1.52 mmol of H2/g of substrate, respectively. The average H2 yield from hydrolysates, obtained from a pretreatment step and enzymatic hydrolysis (3.78 ± 1.92 mmol of H2/g), was lower compared with the yield of substrates pretreated by biological methods only, demonstrating that it is important to avoid the formation of inhibitors generated by chemical pretreatments. Based on this review, exploring other microorganisms and optimizing the pretreatment and hydrolysis conditions can make the use of lignocellulosic substrates a sustainable way to produce H2. PMID:26273246

  13. Enhanced substrate conversion effiency of fermentation processes

    OpenAIRE

    Sanders, J.P.M.; Weusthuis, R.A.; Mooibroek, H.

    2008-01-01

    The present invention relates to the field of fermentation technology. In particular the invention relates to fermentation processes for the production of a first and a second fermentation product by a single production organism wherein the first product is in a more reduced state than the substrate and the second fermentation product is in a more oxidised state than the substrate yet in a less oxidised state than the final oxidation product CO2, such that the concurrent synthesis of the firs...

  14. Substrates and method for determining enzymes

    Science.gov (United States)

    Smith, R.E.; Bissell, E.R.

    1981-10-13

    A method is disclosed for determining the presence of an enzyme in a biological fluid, which includes the steps of contacting the fluid with a synthetic chromogenic substrate, which is an amino acid derivative of 7-amino-4-trifluoromethylcoumarin; incubating the substrate-containing fluid to effect enzymatic hydrolysis; and fluorometrically determining the presence of the free 7-amino-4-trifluoromethylcoumarin chromophore in the hydrolyzate. No Drawings

  15. Alternative substrates for higher mushrooms mycelia cultivation

    Directory of Open Access Journals (Sweden)

    TETIANA KRUPODOROVA

    2015-12-01

    Full Text Available Cultivation of 29 species of higher mushroom mycelia on alternative substrates – wastes of Ukrainian oil-fat industry, has been investigated. The amount of mushroom mycelia obtaining on 12 investigated substrates varied significantly, from 1.0 g/L to 22.9 g/L on the 14th day of cultivation. The superficial cultivation adopted in this study allows for easy to choose appropriate medium (substrate for mycelia production. Alternative substrates (compared to glucose-peptone-yeast medium were selected for all studied species, from soybean cake – most suitable for the mycelial growth of 24 species, to walnut cake − suitable only for 2 species. The utilization of substrates has been evaluated by biological efficiency. The best index of biological efficiency varied from 19.0% to 41.6% depending on the mushroom species. It was established high biological efficiency of mycelia cultivation on substrates: wheat seed cake – Pleurotus djamor, Lyophyllum shimeji, Crinipellis schevczenkovi, Phellinus igniarius, Spongipellis litschaueri; oat seed cake – Ganoderma applanatum and G. lucidum; soybean cake – Hohenbuehelia myxotricha, Trametes versicolor, Morchella esculenta, Cordyceps sinensis, C. militaris, and Agrocybe aegerita; rape seed cake – Auriporia aurea; camelina seed cake – Fomes fomentarius. The cultivation of these species are perspective as a biotechnological process of agricultural wastes converted into mycelia, which could be used in different forms of products with therapeutic action: powder or tablets nutraceuticals or ingredients for functional foods.

  16. Spectrophotometric determination of substrate-borne polyacrylamide.

    Science.gov (United States)

    Lu, Jianhang; Wu, Laosheng

    2002-08-28

    Polyacrylamides (PAMs) have wide application in many industries and in agriculture. Scientific research and industrial applications manifested a need for a method that can quantify substrate-borne PAM. The N-bromination method (a PAM analytical technique based on N-bromination of amide groups and spectrophotometric determination of the formed starch-triiodide complex), which was originally developed for determining PAM in aqueous solutions, was modified to quantify substrate-borne PAM. In the modified method, the quantity of substrate-borne PAM was converted to a concentration of starch-triiodide complex in aqueous solution that was then measured by spectrophotometry. The method sensitivity varied with substrates due to sorption of reagents and reaction intermediates on the substrates. Therefore, separate calibration for each substrate was required. Results from PAM samples in sand, cellulose, organic matter burnt soils, and clay minerals showed that this method had good accuracy and reproducibility. The PAM recoveries ranged from 95.8% to 103.7%, and the relative standard deviations (n = 4) were application and facilitating PAM-related research.

  17. Manufacturing Process for OLED Integrated Substrate

    Energy Technology Data Exchange (ETDEWEB)

    Hung, Cheng-Hung [Vitro Flat Glass LLC, Cheswick, PA (United States). Glass Technology Center

    2017-03-31

    The main objective of this project was to develop a low-cost integrated substrate for rigid OLED solid-state lighting produced at a manufacturing scale. The integrated substrates could include combinations of soda lime glass substrate, light extraction layer, and an anode layer (i.e., Transparent Conductive Oxide, TCO). Over the 3+ year course of the project, the scope of work was revised to focus on the development of a glass substrates with an internal light extraction (IEL) layer. A manufacturing-scale float glass on-line particle embedding process capable of producing an IEL glass substrate having a thickness of less than 1.7mm and an area larger than 500mm x 400mm was demonstrated. Substrates measuring 470mm x 370mm were used in the OLED manufacturing process for fabricating OLED lighting panels in single pixel devices as large as 120.5mm x 120.5mm. The measured light extraction efficiency (calculated as external quantum efficiency, EQE) for on-line produced IEL samples (>50%) met the project’s initial goal.

  18. Use of waste material in cultivation substrates

    Directory of Open Access Journals (Sweden)

    Petr Salaš

    2004-01-01

    Full Text Available Gardeners' practical experience and experimental work prove the affirmation that the used substrate is a very important base for the production of quality nursery products. It is important to emphasis the complexity and synergy of all factors influencing the ecosystem and there mutual relations. Physical, chemical and biological properties do not separately affect the growth and development of plants. In addition, the relations are not statical but differ in relation with other factors changes. This article is dealing with the possibility to use waste material from timber processing in cultivation substrates. The large scale use of such substrates would enable people to reach a relative independence from peat substrates, of which the global reserve is gradually decreasing.Our research activities focus on the use of bark. The basic problems of a bark substrate are easy dehydration and unbalanced nutrition of trees and shrubs. The suggested and experimented cultivation technology solves these problems. It is based on the cultivation of woody species in bark substrates, using modern irrigation systems, slow release fertilisers (Silvamix Forte and special soil conditioners (TerraCottem. This technology was tested on the following species of trees and shrubs: Malus and Buxus.

  19. Nanomechanics of hard films on compliant substrates.

    Energy Technology Data Exchange (ETDEWEB)

    Reedy, Earl David, Jr. (Sandia National Laboratories, Albuquerque, NM); Emerson, John Allen (Sandia National Laboratories, Albuquerque, NM); Bahr, David F. (Washington State University, Pullman, WA); Moody, Neville Reid; Zhou, Xiao Wang; Hales, Lucas (University of Minnesota, Minneapolis, MN); Adams, David Price (Sandia National Laboratories, Albuquerque, NM); Yeager,John (Washington State University, Pullman, WA); Nyugen, Thao D. (Johns Hopkins University, Baltimore, MD); Corona, Edmundo (Sandia National Laboratories, Albuquerque, NM); Kennedy, Marian S. (Clemson University, Clemson, SC); Cordill, Megan J. (Erich Schmid Institute, Leoben, Austria)

    2009-09-01

    Development of flexible thin film systems for biomedical, homeland security and environmental sensing applications has increased dramatically in recent years [1,2,3,4]. These systems typically combine traditional semiconductor technology with new flexible substrates, allowing for both the high electron mobility of semiconductors and the flexibility of polymers. The devices have the ability to be easily integrated into components and show promise for advanced design concepts, ranging from innovative microelectronics to MEMS and NEMS devices. These devices often contain layers of thin polymer, ceramic and metallic films where differing properties can lead to large residual stresses [5]. As long as the films remain substrate-bonded, they may deform far beyond their freestanding counterpart. Once debonded, substrate constraint disappears leading to film failure where compressive stresses can lead to wrinkling, delamination, and buckling [6,7,8] while tensile stresses can lead to film fracture and decohesion [9,10,11]. In all cases, performance depends on film adhesion. Experimentally it is difficult to measure adhesion. It is often studied using tape [12], pull off [13,14,15], and peel tests [16,17]. More recent techniques for measuring adhesion include scratch testing [18,19,20,21], four point bending [22,23,24], indentation [25,26,27], spontaneous blisters [28,29] and stressed overlayers [7,26,30,31,32,33]. Nevertheless, sample design and test techniques must be tailored for each system. There is a large body of elastic thin film fracture and elastic contact mechanics solutions for elastic films on rigid substrates in the published literature [5,7,34,35,36]. More recent work has extended these solutions to films on compliant substrates and show that increasing compliance markedly changes fracture energies compared with rigid elastic solution results [37,38]. However, the introduction of inelastic substrate response significantly complicates the problem [10,39,40]. As

  20. Extensions of the Stoney formula for substrate curvature to configurations with thin substrates or large deformations

    International Nuclear Information System (INIS)

    Freund, L.B.; Floro, J.A.; Chason, E.

    1999-01-01

    Two main assumptions which underlie the Stoney formula relating substrate curvature to mismatch strain in a bonded thin film are that the film is very thin compared to the substrate, and the deformations are infinitesimally small. Expressions for the curvature - strain relationship are derived for cases in which these assumptions are relaxed, thereby providing a basis for interpretation of experimental observations for a broader class of film - substrate configurations. copyright 1999 American Institute of Physics

  1. Decoding P4-ATPase substrate interactions.

    Science.gov (United States)

    Roland, Bartholomew P; Graham, Todd R

    Cellular membranes display a diversity of functions that are conferred by the unique composition and organization of their proteins and lipids. One important aspect of lipid organization is the asymmetric distribution of phospholipids (PLs) across the plasma membrane. The unequal distribution of key PLs between the cytofacial and exofacial leaflets of the bilayer creates physical surface tension that can be used to bend the membrane; and like Ca 2+ , a chemical gradient that can be used to transduce biochemical signals. PL flippases in the type IV P-type ATPase (P4-ATPase) family are the principle transporters used to set and repair this PL gradient and the asymmetric organization of these membranes are encoded by the substrate specificity of these enzymes. Thus, understanding the mechanisms of P4-ATPase substrate specificity will help reveal their role in membrane organization and cell biology. Further, decoding the structural determinants of substrate specificity provides investigators the opportunity to mutationally tune this specificity to explore the role of particular PL substrates in P4-ATPase cellular functions. This work reviews the role of P4-ATPases in membrane biology, presents our current understanding of P4-ATPase substrate specificity, and discusses how these fundamental aspects of P4-ATPase enzymology may be used to enhance our knowledge of cellular membrane biology.

  2. Substrate-Directed Catalytic Selective Chemical Reactions.

    Science.gov (United States)

    Sawano, Takahiro; Yamamoto, Hisashi

    2018-05-04

    The development of highly efficient reactions at only the desired position is one of the most important subjects in organic chemistry. Most of the reactions in current organic chemistry are reagent- or catalyst-controlled reactions, and the regio- and stereoselectivity of the reactions are determined by the inherent nature of the reagent or catalyst. In sharp contrast, substrate-directed reaction determines the selectivity of the reactions by the functional group on the substrate and can strictly distinguish sterically and electronically similar multiple reaction sites in the substrate. In this Perspective, three topics of substrate-directed reaction are mainly reviewed: (1) directing group-assisted epoxidation of alkenes, (2) ring-opening reactions of epoxides by various nucleophiles, and (3) catalytic peptide synthesis. Our newly developed synthetic methods with new ligands including hydroxamic acid derived ligands realized not only highly efficient reactions but also pinpointed reactions at the expected position, demonstrating the substrate-directed reaction as a powerful method to achieve the desired regio- and stereoselective functionalization of molecules from different viewpoints of reagent- or catalyst-controlled reactions.

  3. Multifunctionality is affected by interactions between green roof plant species, substrate depth, and substrate type.

    Science.gov (United States)

    Dusza, Yann; Barot, Sébastien; Kraepiel, Yvan; Lata, Jean-Christophe; Abbadie, Luc; Raynaud, Xavier

    2017-04-01

    Green roofs provide ecosystem services through evapotranspiration and nutrient cycling that depend, among others, on plant species, substrate type, and substrate depth. However, no study has assessed thoroughly how interactions between these factors alter ecosystem functions and multifunctionality of green roofs. We simulated some green roof conditions in a pot experiment. We planted 20 plant species from 10 genera and five families (Asteraceae, Caryophyllaceae, Crassulaceae, Fabaceae, and Poaceae) on two substrate types (natural vs. artificial) and two substrate depths (10 cm vs. 30 cm). As indicators of major ecosystem functions, we measured aboveground and belowground biomasses, foliar nitrogen and carbon content, foliar transpiration, substrate water retention, and dissolved organic carbon and nitrates in leachates. Interactions between substrate type and depth strongly affected ecosystem functions. Biomass production was increased in the artificial substrate and deeper substrates, as was water retention in most cases. In contrast, dissolved organic carbon leaching was higher in the artificial substrates. Except for the Fabaceae species, nitrate leaching was reduced in deep, natural soils. The highest transpiration rates were associated with natural soils. All functions were modulated by plant families or species. Plant effects differed according to the observed function and the type and depth of the substrate. Fabaceae species grown on natural soils had the most noticeable patterns, allowing high biomass production and high water retention but also high nitrate leaching from deep pots. No single combination of factors enhanced simultaneously all studied ecosystem functions, highlighting that soil-plant interactions induce trade-offs between ecosystem functions. Substrate type and depth interactions are major drivers for green roof multifunctionality.

  4. Substrate recognition by ribonucleoprotein ribonuclease MRP.

    Science.gov (United States)

    Esakova, Olga; Perederina, Anna; Quan, Chao; Berezin, Igor; Krasilnikov, Andrey S

    2011-02-01

    The ribonucleoprotein complex ribonuclease (RNase) MRP is a site-specific endoribonuclease essential for the survival of the eukaryotic cell. RNase MRP closely resembles RNase P (a universal endoribonuclease responsible for the maturation of the 5' ends of tRNA) but recognizes distinct substrates including pre-rRNA and mRNA. Here we report the results of an in vitro selection of Saccharomyces cerevisiae RNase MRP substrates starting from a pool of random sequences. The results indicate that RNase MRP cleaves single-stranded RNA and is sensitive to sequences in the immediate vicinity of the cleavage site requiring a cytosine at the position +4 relative to the cleavage site. Structural implications of the differences in substrate recognition by RNases P and MRP are discussed.

  5. Rice Seedling Substrate Produced by Coal Gangue

    Directory of Open Access Journals (Sweden)

    SHAO Yu-fei

    2017-10-01

    Full Text Available Peats are the mostly used material in making rice seedling substrate. However, mining peats could cause environmental problems. In order to reduce or replace peats in rice seedling substrate industry, this paper studied suitable way to configure rice seedling. The coal gangue was used to experiment cultivating rice. Four rice seeding experiments were carried out based on physical and chemical properties of materials attributes. The results showed:(1 Coal gangue was feasible for rice seedling; (2 The maximum adding amount of coal gangue was 80%(volume ratio though the coal gangue need to be activated; (3 In the case of no activated treatment only 38%(volume ratio of coal gangue could be added to the substrate.

  6. Passivation coating for flexible substrate mirrors

    Science.gov (United States)

    Tracy, C. Edwin; Benson, David K.

    1990-01-01

    A protective diffusion barrier for metalized mirror structures is provided by a layer or coating of silicon nitride which is a very dense, transparent, dielectric material that is impervious to water, alkali, and other impurities and corrosive substances that typically attack the metal layers of mirrors and cause degradation of the mirrors' reflectivity. The silicon nitride layer can be deposited on the substrate before metal deposition thereon to stabilize the metal/substrate interface, and it can be deposited over the metal to encapsulate it and protect the metal from corrosion or other degradation. Mirrors coated with silicon nitride according to this invention can also be used as front surface mirrors. Also, the silver or other reflective metal layer on mirrors comprising thin, lightweight, flexible substrates of metal or polymer sheets coated with glassy layers can be protected with silicon nitride according to this invention.

  7. Plasma jet printing for flexible substrates

    Energy Technology Data Exchange (ETDEWEB)

    Gandhiraman, Ram P.; Singh, Eric; Diaz-Cartagena, Diana C.; Koehne, Jessica; Meyyappan, M. [Center for Nanotechnology, NASA Ames Research Center, Moffett Field, California 94035 (United States); Nordlund, Dennis [Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California 94025 (United States)

    2016-03-21

    Recent interest in flexible electronics and wearable devices has created a demand for fast and highly repeatable printing processes suitable for device manufacturing. Robust printing technology is critical for the integration of sensors and other devices on flexible substrates such as paper and textile. An atmospheric pressure plasma-based printing process has been developed to deposit different types of nanomaterials on flexible substrates. Multiwalled carbon nanotubes were deposited on paper to demonstrate site-selective deposition as well as direct printing without any type of patterning. Plasma-printed nanotubes were compared with non-plasma-printed samples under similar gas flow and other experimental conditions and found to be denser with higher conductivity. The utility of the nanotubes on the paper substrate as a biosensor and chemical sensor was demonstrated by the detection of dopamine, a neurotransmitter, and ammonia, respectively.

  8. Printed electronic on flexible and glass substrates

    Science.gov (United States)

    Futera, Konrad; Jakubowska, Małgorzata; Kozioł, Grażyna

    2010-09-01

    Organic electronics is a platform technology that enables multiple applications based on organic electronics but varied in specifications. Organic electronics is based on the combination of new materials and cost-effective, large area production processes that provide new fields of application. Organic electronic by its size, weight, flexibility and environmental friendliness electronics enables low cost production of numerous electrical components and provides for such promising fields of application as: intelligent packaging, low cost RFID, flexible solar cells, disposable diagnostic devices or games, and printed batteries [1]. The paper presents results of inkjetted electronics elements on flexible and glass substrates. The investigations was target on characterizing shape, surface and geometry of printed structures. Variety of substrates were investigated, within some, low cost, non specialized substrate, design for other purposes than organic electronic.

  9. Integration substrate with a ultra-high-density capacitor and a through-substrate via

    NARCIS (Netherlands)

    2008-01-01

    An integration substrate for a system in package comprises a through-substrate via and a trench capacitor wherein with a trench filling that includes at least 4 elec. conductive capacitor-electrode layers in an alternating arrangement with dielec. layers. The capacitor-electrode layers are

  10. Integration substrate with a ultra-high-density capacitor and a through-substrate via

    NARCIS (Netherlands)

    Klootwijk, J.H.; Roozeboom, F.; Ruigrok, J.J.M.; Reefman, D.

    2014-01-01

    An integration substrate for a system in package comprises a through-substrate via and a trench capacitor wherein with a trench filling that includes at least four electrically conductive capacitor-electrode layers in an alternating arrangement with dielectric layers. --The capacitor-electrode

  11. Equilibrium Droplets on Deformable Substrates: Equilibrium Conditions.

    Science.gov (United States)

    Koursari, Nektaria; Ahmed, Gulraiz; Starov, Victor M

    2018-05-15

    Equilibrium conditions of droplets on deformable substrates are investigated, and it is proven using Jacobi's sufficient condition that the obtained solutions really provide equilibrium profiles of both the droplet and the deformed support. At the equilibrium, the excess free energy of the system should have a minimum value, which means that both necessary and sufficient conditions of the minimum should be fulfilled. Only in this case, the obtained profiles provide the minimum of the excess free energy. The necessary condition of the equilibrium means that the first variation of the excess free energy should vanish, and the second variation should be positive. Unfortunately, the mentioned two conditions are not the proof that the obtained profiles correspond to the minimum of the excess free energy and they could not be. It is necessary to check whether the sufficient condition of the equilibrium (Jacobi's condition) is satisfied. To the best of our knowledge Jacobi's condition has never been verified for any already published equilibrium profiles of both the droplet and the deformable substrate. A simple model of the equilibrium droplet on the deformable substrate is considered, and it is shown that the deduced profiles of the equilibrium droplet and deformable substrate satisfy the Jacobi's condition, that is, really provide the minimum to the excess free energy of the system. To simplify calculations, a simplified linear disjoining/conjoining pressure isotherm is adopted for the calculations. It is shown that both necessary and sufficient conditions for equilibrium are satisfied. For the first time, validity of the Jacobi's condition is verified. The latter proves that the developed model really provides (i) the minimum of the excess free energy of the system droplet/deformable substrate and (ii) equilibrium profiles of both the droplet and the deformable substrate.

  12. Ferromagnetic film on a superconducting substrate

    Energy Technology Data Exchange (ETDEWEB)

    Bulaevskii, L. N.; Chudnovsky, E. M.

    2001-01-01

    We study the equilibrium domain structure and magnetic flux around a ferromagnetic film with perpendicular magnetization M{sub 0} on a superconducting (SC) substrate. At 4{pi}M{sub 0}substrate, l{sub N}/4{pi}{lambda}{sub L}>>1; {lambda}{sub L} being the London penetration length.

  13. Ferromagnetic film on a superconducting substrate

    International Nuclear Information System (INIS)

    Bulaevskii, L. N.; Chudnovsky, E. M.

    2001-01-01

    We study the equilibrium domain structure and magnetic flux around a ferromagnetic film with perpendicular magnetization M 0 on a superconducting (SC) substrate. At 4πM 0 c1 the SC is in the Meissner state and the equilibrium domain width in the film, l, scales as (l/4πλ L )=(l N /4πλ L ) 2/3 with the domain width on a normal (nonsuperconducting) substrate, l N /4πλ L >>1; λ L being the London penetration length

  14. Protein degradation: recognition of ubiquitinylated substrates

    DEFF Research Database (Denmark)

    Hartmann-Petersen, Rasmus; Gordon, Colin

    2004-01-01

    A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role.......A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role....

  15. Bayesian inference of substrate properties from film behavior

    International Nuclear Information System (INIS)

    Aggarwal, R; Demkowicz, M J; Marzouk, Y M

    2015-01-01

    We demonstrate that by observing the behavior of a film deposited on a substrate, certain features of the substrate may be inferred with quantified uncertainty using Bayesian methods. We carry out this demonstration on an illustrative film/substrate model where the substrate is a Gaussian random field and the film is a two-component mixture that obeys the Cahn–Hilliard equation. We construct a stochastic reduced order model to describe the film/substrate interaction and use it to infer substrate properties from film behavior. This quantitative inference strategy may be adapted to other film/substrate systems. (paper)

  16. Inverter power module with distributed support for direct substrate cooling

    Science.gov (United States)

    Miller, David Harold [San Pedro, CA; Korich, Mark D [Chino Hills, CA; Ward, Terence G [Redondo Beach, CA; Mann, Brooks S [Redondo Beach, CA

    2012-08-21

    Systems and/or methods are provided for an inverter power module with distributed support for direct substrate cooling. An inverter module comprises a power electronic substrate. A first support frame is adapted to house the power electronic substrate and has a first region adapted to allow direct cooling of the power electronic substrate. A gasket is interposed between the power electronic substrate and the first support frame. The gasket is configured to provide a seal between the first region and the power electronic substrate. A second support frame is adapted to house the power electronic substrate and joined to the first support frame to form the seal.

  17. Method for producing substrates for superconducting layers

    DEFF Research Database (Denmark)

    2013-01-01

    There is provided a method for producing a substrate (600) suitable for supporting an elongated superconducting element, wherein, e.g., a deformation process is utilized in order to form disruptive strips in a layered solid element, and where etching is used to form undercut volumes (330, 332...

  18. Method for Producing Substrates for Superconducting Layers

    DEFF Research Database (Denmark)

    2015-01-01

    There is provided a method for producing a substrate suitable for supporting an elongated superconducting element, wherein one or more elongated strips of masking material are placed on a solid element (202) so as to form one or more exposed elongated areas being delimited on one or two sides...

  19. Mass spectrometry-assisted protease substrate screening

    DEFF Research Database (Denmark)

    Schlüter, Hartmut; Rykl, Jana; Thiemann, Joachim

    2007-01-01

    -phase chromatography they are analyzed by tandem mass spectrometry and the substrates identified by database searching. The proof of principle in this study is demonstrated by incubating immobilized human plasma proteins with thrombin and by identifying by tandem mass spectrometry the fibrinopeptides, released...

  20. Biodegradation of hydrocarbons exploiting spent substrate from ...

    African Journals Online (AJOL)

    The aim of this study was to evaluate the mushroom substrate of P. ostreatus in a microcosm for the bioremediation of an agricultural soil contaminated with diesel. We evaluated the participation of microbial populations and specific enzymatic lacasses, manganese peroxidases, versatile peroxidases, veratryl alcohol ...

  1. Microbial reduction of uranium using cellulosic substrates

    International Nuclear Information System (INIS)

    Thombre, M.S.; Thomson, B.M.; Barton, L.L.

    1996-01-01

    Previous work at the University of New Mexico and elsewhere has shown that sulfate-reducing bacteria are capable of reducing uranium from the soluble +6 oxidation state to the insoluble +4 oxidation state. This chemistry forms the basis of a proposed ground water remediation strategy in which microbial reduction would be used to immobilize soluble uranium. One such system would consist of a subsurface permeable barrier which would stimulate microbial growth resulting in the reduction of sulfate and nitrate and immobilization of metals while permitting the unhindered flow of ground water through it. This research investigated some of the engineering considerations associated with a microbial reducing barrier such as identifying an appropriate biological substrate, estimating the rate of substrate utilization, and identifying the final fate of the contaminants concentrated in the barrier matrix. The performance of batch reactors and column systems that treated simulated plume water was evaluated using cellulose, wheat straw, alfalfa hay, sawdust, and soluble starch as substrates. The concentrations of sulfate, nitrate, and U(VI) were monitored over time. Precipitates from each system were collected, and the precipitated U(IV) was determined to be crystalline UO 2(s) by x-ray diffraction. The results of this study support the proposed use of cellulosic substrates as candidate barrier materials

  2. A Spectral Emissivity Library of Spoil Substrates

    Czech Academy of Sciences Publication Activity Database

    Pivovarník, Marek; Pikl, Miroslav; Frouz, J.; Zemek, František; Kopačková, V.; Notesco, G.; Ben Dor, E.

    2016-01-01

    Roč. 1, č. 2 (2016) E-ISSN 2306-5729 R&D Projects: GA MŠk(CZ) LO1415 Institutional support: RVO:86652079 Keywords : post-mining sites * spectral emissivity * spectral library * spoil substrates Subject RIV: EH - Ecology, Behaviour OBOR OECD: Environmental sciences (social aspects to be 5.7)

  3. Modification of chitin as substrates for chitinase

    African Journals Online (AJOL)

    sunny t

    2015-05-06

    May 6, 2015 ... Enzymes are able to bind to their substrates specifically at the active site. The proximity and ... the presence of chitin as a carbon source (Chernin et al.,. 1998). ... Possible rearrangement of chitin structure ... and form larger granules. .... Medium for Enumeration of Actinomycetes in Water and Soil. Appl.

  4. Buckling Behavior of Substrate Supported Graphene Sheets

    Directory of Open Access Journals (Sweden)

    Kuijian Yang

    2016-01-01

    Full Text Available The buckling of graphene sheets on substrates can significantly degrade their performance in materials and devices. Therefore, a systematic investigation on the buckling behavior of monolayer graphene sheet/substrate systems is carried out in this paper by both molecular mechanics simulations and theoretical analysis. From 70 simulation cases of simple-supported graphene sheets with different sizes under uniaxial compression, two different buckling modes are investigated and revealed to be dominated by the graphene size. Especially, for graphene sheets with length larger than 3 nm and width larger than 1.1 nm, the buckling mode depends only on the length/width ratio. Besides, it is revealed that the existence of graphene substrate can increase the critical buckling stress and strain to 4.39 N/m and 1.58%, respectively, which are about 10 times those for free-standing graphene sheets. Moreover, for graphene sheets with common size (longer than 20 nm, both theoretical and simulation results show that the critical buckling stress and strain are dominated only by the adhesive interactions with substrate and independent of the graphene size. Results in this work provide valuable insight and guidelines for the design and application of graphene-derived materials and nano-electromechanical systems.

  5. Optical characterization of nanoporous AAO sensor substrate

    Science.gov (United States)

    Kassu, Aschalew; Farley, Carlton W.; Sharma, Anup

    2014-05-01

    Nanoporous anodic aluminum oxide (AAO) has been investigated as an ideal and cost-effective chemical and biosensing platform. In this paper, we report the optical properties of periodic 100 micron thick nanoporous anodic alumina membranes with uniform and high density cylindrical pores penetrating the entire thickness of the substrate, ranging in size from 18 nm to 150 nm in diameter and pore periods from 44 nm to 243 nm. The surface geometry of the top and bottom surface of each membrane is studied using atomic force microscopy. The optical properties including transmittance, reflectance, and absorbance spectra on both sides of each substrate are studied and found to be symmetrical. It is observed that, as the pore size increases, the peak resonance intensity in transmittance decreases and in absorbance increases. The effects of the pore sizes on the optical properties of the bare nanoporous membranes and the benefit of using arrays of nanohole arrays with varying hole size and periodicity as a chemical sensing platform is also discussed. To characterize the optical sensing technique, transmittance and reflectance measurements of various concentrations of a standard chemical adsorbed on the bare nanoporous substrates are investigated. The preliminary results presented here show variation in transmittance and reflectance spectra with the concentration of the chemical used or the amount of the material adsorbed on the surface of the substrate.

  6. Enhanced substrate conversion efficiency of fermentation processes

    NARCIS (Netherlands)

    Sanders, J.P.M.; Weusthuis, R.A.; Mooibroek, H.

    2006-01-01

    The present invention relates to the field of fermentation technology. In particular the invention relates to fermentation processes for the production of a first and a second fermentation product by a single production organism wherein the first product is in a more reduced state than the substrate

  7. Enhanced substrate conversion effiency of fermentation processes

    NARCIS (Netherlands)

    Sanders, J.P.M.; Weusthuis, R.A.; Mooibroek, H.

    2008-01-01

    The present invention relates to the field of fermentation technology. In particular the invention relates to fermentation processes for the production of a first and a second fermentation product by a single production organism wherein the first product is in a more reduced state than the substrate

  8. Optimizing substrate for sulfate-reducing bacteria

    International Nuclear Information System (INIS)

    Chang, L.K.; Updegraff, D.M.; Wildeman, T.R.

    1991-01-01

    Microbial sulfate reduction followed by sulfide precipitation effectively removes heavy metals from wastewaters. The substrate in the anaerobic zone in a constructed wetland can be designed to emphasize this removal process. This group of bacteria requires CH 2 O, P, N, and SO 4 =, reducing conditions, and pH range of 5-9 (pH=7 is optimum). The objective of this study was to find an inexpensive source of nutrients that would give the best initial production of sulfide and make a good wetland substrate. All tested materials contain sufficient P and N; mine drainage provides sulfate. Thus, tests focused on finding organic material that provides the proper nutrients and does not cause the culture to fall below pH of 5. Among chemical nutrients, sodium lactate combined with (NH 4 ) 2 HPO 4 were the only compounds that produced sulfide after 11 days. Among complex nutrients, only cow manure produced sulfide after 26 days. Among complex carbohydrates, cracked corn and raw rice produced sulfide after 10 days. Most substrates failed to produce sulfide because anaerobic fermentation reduced the pH below 5. Presently, cracked corn is the best candidate for a substrate. Five grams of cow manure produced 0.14 millimole of sulfide whereas 0.1 g of cracked corn produced 0.22 millimole

  9. Influence of substrate modulus on gecko adhesion

    Science.gov (United States)

    Klittich, Mena R.; Wilson, Michael C.; Bernard, Craig; Rodrigo, Rochelle M.; Keith, Austin J.; Niewiarowski, Peter H.; Dhinojwala, Ali

    2017-03-01

    The gecko adhesion system fascinates biologists and materials scientists alike for its strong, reversible, glue-free, dry adhesion. Understanding the adhesion system’s performance on various surfaces can give clues as to gecko behaviour, as well as towards designing synthetic adhesive mimics. Geckos encounter a variety of surfaces in their natural habitats; tropical geckos, such as Gekko gecko, encounter hard, rough tree trunks as well as soft, flexible leaves. While gecko adhesion on hard surfaces has been extensively studied, little work has been done on soft surfaces. Here, we investigate for the first time the influence of macroscale and nanoscale substrate modulus on whole animal adhesion on two different substrates (cellulose acetate and polydimethylsiloxane) in air and find that across 5 orders of magnitude in macroscale modulus, there is no change in adhesion. On the nanoscale, however, gecko adhesion is shown to depend on substrate modulus. This suggests that low surface-layer modulus may inhibit the gecko adhesion system, independent of other influencing factors such as macroscale composite modulus and surface energy. Understanding the limits of gecko adhesion is vital for clarifying adhesive mechanisms and in the design of synthetic adhesives for soft substrates (including for biomedical applications and wearable electronics).

  10. Biochemicalmethane potential (BMP) of solid organic substrates

    DEFF Research Database (Denmark)

    Raposo, F.; Fernández-Cegrí, V.; de la Rubia, M.A.

    2011-01-01

    BACKGROUND: This paper describes results obtained for different participating research groups in an interlaboratory study related to biochemical methane potential (BMP). In this research work, all experimental conditions influencing the test such as inoculum, substrate characteristics and experim...... significantly according to the experimental approaches....

  11. Integrated Plastic Substrates for OLED Lighting

    Energy Technology Data Exchange (ETDEWEB)

    Gaynor, Whitney

    2015-08-01

    OLED lighting has immense potential as aesthetically pleasing, energy-efficient general illumination. Unlike other light sources, such as incandescents, fluorescents, and inorganic LEDs, OLEDs naturally emit over a large-area surface. They are glare free, do not need to be shaded, and are cool to the touch, requiring no heatsink. The best efficiencies and lifetimes reported are on par with or better than current forms of illumination. However, the cost for OLED lighting remains high – so much so that these products are not market competitive and there is very low consumer demand. We believe that flexible, plastic-based devices will highlight the advantages of aesthetically-pleasing OLED lighting systems while paving the way for lowering both materials and manufacturing costs. These flexible devices require new development in substrate and support technology, which was the focus of the work reported here. The project team, led by Sinovia Technologies, has developed integrated plastic substrates to serve as supports for flexible OLED lighting. The substrates created in this project would enable large-area, flexible devices and are specified to perform three functions. They include a barrier to protect the OLED from moisture and oxygen-related degradation, a smooth, highly conductive transparent electrode to enable large-area device operation, and a light scattering layer to improve emission efficiency. Through the course of this project, integrated substrates were fabricated, characterized, evaluated for manufacturing feasibility and cost, and used in white OLED demonstrations to test their impact on flexible OLED lighting. Our integrated substrates meet or exceed the DOE specifications for barrier performance in water vapor and oxygen transport rates, as well as the transparency and conductivity of the anode film. We find that these integrated substrates can be manufactured in a completely roll-to-roll, high throughput process and have developed and demonstrated

  12. Silvering substrates after CO2 snow cleaning

    Science.gov (United States)

    Zito, Richard R.

    2005-09-01

    There have been some questions in the astronomical community concerning the quality of silver coatings deposited on substrates that have been cleaned with carbon dioxide snow. These questions center around the possible existence of carbonate ions left behind on the substrate by CO2. Such carbonate ions could react with deposited silver to produce insoluble silver carbonate, thereby reducing film adhesion and reflectivity. Carbonate ions could be produced from CO2 via the following mechanism. First, during CO2 snow cleaning, a small amount of moisture can condense on a surface. This is especially true if the jet of CO2 is allowed to dwell on one spot. CO2 gas can dissolve in this moisture, producing carbonic acid, which can undergo two acid dissociations to form carbonate ions. In reality, it is highly unlikely that charged carbonate ions will remain stable on a substrate for very long. As condensed water evaporates, Le Chatelier's principle will shift the equilibrium of the chain of reactions that produced carbonate back to CO2 gas. Furthermore, the hydration of CO2 reaction of CO2 with H20) is an extremely slow process, and the total dehydrogenation of carbonic acid is not favored. Living tissues that must carry out the equilibration of carbonic acid and CO2 use the enzyme carbonic anhydrase to speed up the reaction by a factor of one million. But no such enzymatic action is present on a clean mirror substrate. In short, the worst case analysis presented below shows that the ratio of silver atoms to carbonate radicals must be at least 500 million to one. The results of chemical tests presented here support this view. Furthermore, film lift-off tests, also presented in this report, show that silver film adhesion to fused silica substrates is actually enhanced by CO2 snow cleaning.

  13. Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate.

    Science.gov (United States)

    Johnson, Joseph L; Cusack, Bernadette; Davies, Matthew P; Fauq, Abdul; Rosenberry, Terrone L

    2003-05-13

    Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3-(acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild-type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate

  14. Multifunctional epitaxial systems on silicon substrates

    Energy Technology Data Exchange (ETDEWEB)

    Singamaneni, Srinivasa Rao, E-mail: ssingam@ncsu.edu [Department of Materials Science and Engineering, North Carolina State University, Raleigh, North Carolina 27695 (United States); Materials Science Division, Army Research Office, Research Triangle Park, North Carolina 27709 (United States); Department of Physics, The University of Texas at El Paso, El Paso, Texas 79968 (United States); Prater, John Thomas [Department of Materials Science and Engineering, North Carolina State University, Raleigh, North Carolina 27695 (United States); Materials Science Division, Army Research Office, Research Triangle Park, North Carolina 27709 (United States); Narayan, Jagdish [Department of Materials Science and Engineering, North Carolina State University, Raleigh, North Carolina 27695 (United States)

    2016-09-15

    Multifunctional heterostructures can exhibit a wide range of functional properties, including colossal magneto-resistance, magnetocaloric, and multiferroic behavior, and can display interesting physical phenomena including spin and charge ordering and strong spin-orbit coupling. However, putting this functionality to work remains a challenge. To date, most of the work reported in the literature has dealt with heterostructures deposited onto closely lattice matched insulating substrates such as DyScO{sub 3}, SrTiO{sub 3} (STO), or STO buffered Si(100) using concepts of lattice matching epitaxy (LME). However, strain in heterostructures grown by LME is typically not fully relaxed and the layers contain detrimental defects such as threading dislocations that can significantly degrade the physical properties of the films and adversely affect the device characteristics. In addition, most of the substrates are incompatible with existing CMOS-based technology, where Si (100) substrates dominate. This review discusses recent advances in the integration of multifunctional oxide and non-oxide materials onto silicon substrates. An alternative thin film growth approach, called “domain matching epitaxy,” is presented which identifies approaches for minimizing lattice strain and unwanted defects in large misfit systems (7%–25% and higher). This approach broadly allows for the integration of multifunctional materials onto silicon substrates, such that sensing, computation, and response functions can be combined to produce next generation “smart” devices. In general, pulsed laser deposition has been used to epitaxially grow these materials, although the concepts developed here can be extended to other deposition techniques, as well. It will be shown that TiN and yttria-stabilized zirconia template layers provide promising platforms for the integration of new functionality into silicon-based computer chips. This review paper reports on a number of thin

  15. Self-Assembled Local Artificial Substrates of GaAs on Si Substrate

    Directory of Open Access Journals (Sweden)

    Frigeri C

    2010-01-01

    Full Text Available Abstract We propose a self-assembling procedure for the fabrication of GaAs islands by Droplet Epitaxy on silicon substrate. Controlling substrate temperature and amount of supplied gallium is possible to tune the base size of the islands from 70 up to 250 nm and the density from 107 to 109 cm−2. The islands show a standard deviation of base size distribution below 10% and their shape evolves changing the aspect ratio from 0.3 to 0.5 as size increases. Due to their characteristics, these islands are suitable to be used as local artificial substrates for the integration of III–V quantum nanostructures directly on silicon substrate.

  16. Dewetting of thin polymer film on rough substrate: II. Experiment

    International Nuclear Information System (INIS)

    Volodin, Pylyp; Kondyurin, Alexey

    2008-01-01

    The theory of the dewetting process developed for a model of substrate-film interaction forces was examined by an experimental investigation of the dewetting process of thin polystyrene (PS) films on chemically etched silicon substrates. In the dependence on PS films thickness and silicon roughness, various situations of dewetting were observed as follows: (i) if the wavelength of the substrate roughness is much larger than the critical spinodal wavelength of a film, then spinodal dewetting of the film is observed; (ii) if the wavelength of the substrate roughness is smaller than the critical wavelength of the film and the substrate roughness is larger in comparison with film thickness, then the dewetting due to substrate roughness is observed and the dewetted film patterns repeat the rough substrate structure; (iii) if the wavelength of the substrate roughness is smaller than the critical wavelength of the film and the substrate roughness is small in comparison with the film thickness, then spinodal dewetting proceeds

  17. Carbon nanotube substrates and catalyzed hot stamp for polishing and patterning the substrates

    Science.gov (United States)

    Wang, Yuhuang [Evanston, IL; Hauge, Robert H [Houston, TX; Schmidt, Howard K [Houston, TX; Kim, Myung Jong [Houston, TX; Kittrell, W Carter [Houston, TX

    2009-09-08

    The present invention is generally directed to catalyzed hot stamp methods for polishing and/or patterning carbon nanotube-containing substrates. In some embodiments, the substrate, as a carbon nanotube fiber end, is brought into contact with a hot stamp (typically at 200-800.degree. C.), and is kept in contact with the hot stamp until the morphology/patterns on the hot stamp have been transferred to the substrate. In some embodiments, the hot stamp is made of material comprising one or more transition metals (Fe, Ni, Co, Pt, Ag, Au, etc.), which can catalyze the etching reaction of carbon with H.sub.2, CO.sub.2, H.sub.2O, and/or O.sub.2. Such methods can (1) polish the carbon nanotube-containing substrate with a microscopically smooth finish, and/or (2) transfer pre-defined patterns from the hot stamp to the substrate. Such polished or patterned carbon nanotube substrates can find application as carbon nanotube electrodes, field emitters, and field emitter arrays for displays and electron sources.

  18. Substrate binding accelerates the conformational transitions and substrate dissociation in multidrug efflux transporter AcrB

    Directory of Open Access Journals (Sweden)

    Beibei eWang

    2015-04-01

    Full Text Available The tripartite efflux pump assembly AcrAB-TolC is the major multidrug resistance transporter in E. coli. The inner membrane transporter AcrB is a homotrimer, energized by the proton movement down the transmembrane electrochemical gradient. The asymmetric crystal structures of AcrB with three monomers in distinct conformational states (access (A, binding (B and extrusion (E support a functional rotating mechanism, in which each monomer of AcrB cycles among the three states in a concerted way. However, the relationship between the conformational changes during functional rotation and drug translocation has not been totally understood. Here, we explored the conformational changes of the AcrB homotrimer during the ABE→BEA transition in different substrate-binding states using targeted MD simulations. It was found that the dissociation of substrate from the distal binding pocket of B monomer is closely related to the concerted conformational changes in the translocation pathway, especially the side chain reorientation of Phe628 and Tyr327. A second substrate binding at the proximal binding pocket of A monomer evidently accelerates the conformational transitions as well as substrate dissociation in B monomer. The acceleration effect of the multi-substrate binding mode provides a molecular explanation for the positive cooperativity observed in the kinetic studies of substrate efflux and deepens our understanding of the functional rotating mechanism of AcrB.

  19. Substrate effect on oxygen reduction electrocatalysis

    International Nuclear Information System (INIS)

    Timperman, L.; Feng, Y.J.; Vogel, W.; Alonso-Vante, N.

    2010-01-01

    The oxygen reduction reaction (ORR) was investigated on carbon (XC-72) supported platinum nanoparticles, generated via the carbonyl chemical route and on oxide composites supported platinum generated via the UV-photo-deposition technique in sulfuric acid medium. The behavior of Pt/C was examined using a careful dosing of the catalyst loading spanning the range from 4.3 to 131 μg cm -2 . The ORR electrochemical response of Pt/C (in line with recent literature data) is put into contrast with the Pt/oxide-composite systems. Our results point out that it is possible to use smaller amounts of catalyst for the ORR when platinum atoms interact with the oxide (anatase) surface of the substrate composite. Evidence of the incipient metal-substrate interaction is discussed in the light of the results of XRD experiments.

  20. Formation of boride layers on steel substrates

    International Nuclear Information System (INIS)

    Stergioudis, G.

    2006-01-01

    Boronizing coatings were prepared by means of pack cementation technique. It was found that using the appropriate substrate and controlling parameters of the boribing process such as boron activity of the mixture, temperature and time of treatment, it is possible to obtain a structure predominantly consisting of the Fe 2 B phase. In the present study low alloy ferritic steels were chosen as substrates. Changing the boron carbide concentration in the mixture and the temperature and time of boronizing process the conditions of the boronizing were altered. As a result the formation of the Fe 2 B phase is enhanced. Characterization of the as-borided steels is discussed based on X-ray diffraction and Curie temperature measurements. (copyright 2006 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  1. Optical substrate materials for synchrotron radiation beamlines

    International Nuclear Information System (INIS)

    Howells, M.R.; Paquin, R.A.

    1997-06-01

    The authors consider the materials choices available for making optical substrates for synchrotron radiation beam lines. They find that currently the optical surfaces can only be polished to the required finish in fused silica and other glasses, silicon, CVD silicon carbide, electroless nickel and 17-4 PH stainless steel. Substrates must therefore be made of one of these materials or of a metal that can be coated with electroless nickel. In the context of material choices for mirrors they explore the issues of dimensional stability, polishing, bending, cooling, and manufacturing strategy. They conclude that metals are best from an engineering and cost standpoint while the ceramics are best from a polishing standpoint. They then give discussions of specific materials as follows: silicon carbide, silicon, electroless nickel, Glidcop trademark, aluminum, precipitation-hardening stainless steel, mild steel, invar and superinvar. Finally they summarize conclusions and propose ideas for further research

  2. Varactor-tuned Substrate Integrated Evanescent Filter

    DEFF Research Database (Denmark)

    Zhurbenko, Vitaliy; Acar, Öncel; Dong, Yunfeng

    Evanescent mode waveguides allow for more compact microwave component design in comparison to the traditional fundamental mode waveguide technology. Evanescent waveguides can be integrated into a dielectric substrate in order to further reduce the mass and volume. Unfortunately, traditional...... realization methods used in the standard evanescent waveguides are often not directly applicable to substrate integrated waveguide (SIW) technology due to dielectric filling and small height of the waveguide. In this work, one of the realization methods of evanescent waveguides using lumped elements...... is considered. In contrast to other methods described in the literature, it avoids etching split ring resonators in the metal layer of the SIW. The filters presented here use varactors as tuning elements. The varactors (as well as DC decoupling circuits) are mounted on the surface of PCB bringing the lower...

  3. Hydrazones as substrates for cycloaddition reactions

    International Nuclear Information System (INIS)

    Belskaya, N P; Eliseeva, A I; Bakulev, V A

    2015-01-01

    The [2+2]-, [4+2]- and [3+2]-cycloaddition reactions of hydrazones and 1,2-diazabuta-1,3-dienes, azomethine imines, nitrile imines and azomethine ylides formed upon hydrazone transformations with dienophiles, dipolarophiles and dienes are considered. The principal issues of structure and reactivity of active substrates and the influence of the reaction conditions and catalysts on the reaction regioselectivity and efficiency are discussed. The bibliography includes 288 references

  4. Purity of targets prepared on Cu substrates

    Science.gov (United States)

    Méens, A.; Rossini, I.; Sens, J. C.

    1993-09-01

    The purity of several elemental self-supporting targets usually prepared by evaporation onto soluble Cu substrates has been studied. The targets were analysed by Rutherford backscattering and instrumental neutron activation analysis. Because of the high percentage of Cu observed in some Si targets, further measurements, including transmission electron microscopy, have been performed on Si targets deposited by e-gun bombardment onto Cu and ion-beam sputtering onto betaine.

  5. Substrate Integrated Evanescent Filters Employing Coaxial Stubs

    DEFF Research Database (Denmark)

    Zhurbenko, Vitaliy

    2015-01-01

    Evanescent mode substrate integrated waveguide (SIW) is one of the promising technologies for design of light-weight low-cost microwave components. Traditional realization methods used in the standard evanescent waveguide technology are often not directly applicable to SIW due to dielectric filli...... of the microwave filter are discussed. The approach is useful in applications where a sharp roll-off at the upper stop-band is required....

  6. Natural cellulose fiber as substrate for supercapacitor.

    Science.gov (United States)

    Gui, Zhe; Zhu, Hongli; Gillette, Eleanor; Han, Xiaogang; Rubloff, Gary W; Hu, Liangbing; Lee, Sang Bok

    2013-07-23

    Cellulose fibers with porous structure and electrolyte absorption properties are considered to be a good potential substrate for the deposition of energy material for energy storage devices. Unlike traditional substrates, such as gold or stainless steel, paper prepared from cellulose fibers in this study not only functions as a substrate with large surface area but also acts as an interior electrolyte reservoir, where electrolyte can be absorbed much in the cellulose fibers and is ready to diffuse into an energy storage material. We demonstrated the value of this internal electrolyte reservoir by comparing a series of hierarchical hybrid supercapacitor electrodes based on homemade cellulose paper or polyester textile integrated with carbon nanotubes (CNTs) by simple solution dip and electrodeposited with MnO2. Atomic layer deposition of Al2O3 onto the fiber surface was used to limit electrolyte absorption into the fibers for comparison. Configurations designed with different numbers of ion diffusion pathways were compared to show that cellulose fibers in paper can act as a good interior electrolyte reservoir and provide an effective pathway for ion transport facilitation. Further optimization using an additional CNT coating resulted in an electrode of paper/CNTs/MnO2/CNTs, which has dual ion diffusion and electron transfer pathways and demonstrated superior supercapacitive performance. This paper highlights the merits of the mesoporous cellulose fibers as substrates for supercapacitor electrodes, in which the water-swelling effect of the cellulose fibers can absorb electrolyte, and the mesoporous internal structure of the fibers can provide channels for ions to diffuse to the electrochemical energy storage materials.

  7. Advanced organics for electronic substrates and packages

    CERN Document Server

    Fletcher, Andrew E

    1992-01-01

    Advanced Organics for Electronic Substrates and Packages provides information on packaging, which is one of the most technologically intensive activities in the electronics industry. The electronics packaging community has realized that while semiconductor devices continue to be improved upon for performance, cost, and reliability, it is the interconnection or packaging of these devices that will limit the performance of the systems. Technology must develop packaging for transistor chips, with high levels of performance and integration providing cooling, power, and interconnection, and yet pre

  8. Flexible SERS Substrates: Challenges and Opportunities

    Science.gov (United States)

    2016-01-28

    are still widely used due to the ease with which silver and gold nanoparticles can be produced. Nanoparticle inks are colloidal suspensions of...interactions between the analyte, silver nanoparticles, and a salt. This system has also been applied to detection of trace antibiotics for food safety...Cleanable SERS Substrates Based on Silver Nanoparticle Decorated Electrospun Nano-fibrous Membranes Chaoyang Jiang Porous electrospun nanofibrous

  9. CONICAL EQUIPOTENTIAL SUBSTRATE FOR LIQUID METAL SOURCES

    OpenAIRE

    Kubby , J.; Siegel , B.

    1986-01-01

    Morphological changes that occur at an ion bombarded surface as a result of erosion by sputtering can be utilized for the machining of cylindrically symmetric submicron structures. Such structuring has produced tungsten field emitters of conical configuration with variable cone half angle. A conical equipotential surface with an included half angle of 49.3° would be a useful emitter substrate for experiments designed to produce an equilibrium conical interface to a conducting fluid in an appl...

  10. Substrate-induced strain in carbon nanodisks

    International Nuclear Information System (INIS)

    Osváth, Z.; Vértesy, Z.; Lábár, J.; Nemes-Incze, P.; Horváth, Z.E.; Biró, L.P.

    2014-01-01

    Graphitic nanodisks of typically 20–50 nm in thickness, produced by the so-called Kvaerner Carbon Black and Hydrogen Process were dispersed on gold substrate and investigated by atomic force microscopy (AFM), field emission scanning electron microscopy (FE-SEM), and confocal Raman spectroscopy. The roughness of the gold surface was drastically changed by annealing at 400 °C. AFM measurements show that this change in the surface roughness induces changes also in the topography of the nanodisks, as they closely follow the corrugation of the gold substrate. This leads to strained nanodisks, which is confirmed also by confocal Raman microscopy. We found that the FE-SEM contrast obtained from the disks depends on the working distance used during the image acquisition by In-lens detection, a phenomenon which we explain by the decrease in the amount of electrons reaching the detector due to diffraction. This process may affect the image contrast in the case of other layered materials, like hexagonal boron nitride, and other planar hybrid nanostructures, too. - Highlights: • Bending of carbon nanodisks is induced by the roughness of the gold substrate. • Confocal Raman microscopy shows a compressive strain induced in the nanodisks. • The electron microscopy contrast of nanodisks depends on the working distance

  11. Liquid crystal displays with plastic substrates

    Science.gov (United States)

    Lueder, Ernst H.

    1998-04-01

    Plastic substrates for the cells of displays exhibit only 1/6 of the weight of glass substrates; they are virtually unbreakable; their flexibility allows the designer to give them a shape suppressing reflections, to realize a display board on a curved surface or meeting the requirements for an appealing styling; displays with plastics are thinner which provides a wider viewing angle. These features render them attractive for displays in portable systems such as mobile phones, pagers, smart cards, personal digital assistants (PDAs) and portable computers. Reflective displays are especially attractive as they don't need a back light. The most important requirements are the protection of plastics against gas permeation and chemical agents, the prevention of layers on plastics to crack or peel off when the plastic is bent and the development of low temperature thin film processes because the plastics, as a rule, only tolerate temperatures below 150 degrees Celsius. Bistable reflective FLC- and PSCT-displays with plastic substrates will be introduced. Special sputtered SiO2-orientation layers preserve the displayed information even if pressure or torsion is applied. MIM-addressed PDLC-displays require additional Al- or Ti-layers which provide the necessary ductility. Sputtered or PECVD-generated TFTs can be fabricated on plastics at temperatures below 150 degrees Celsius.

  12. Nanowire surface fastener fabrication on flexible substrate

    Science.gov (United States)

    Toku, Yuhki; Uchida, Keita; Morita, Yasuyuki; Ju, Yang

    2018-07-01

    The market for wearable devices has increased considerably in recent years. In response to this demand, flexible electronic circuit technology has become more important. The conventional bonding technology in electronic assembly depends on high-temperature processes such as reflow soldering, which result in undesired thermal damages and residual stress at a bonding interface. In addition, it exhibits poor compatibility with bendable or stretchable device applications. Therefore, there is an urgent requirement to attach electronic parts on printed circuit boards with good mechanical and electrical properties at room temperature. Nanowire surface fasteners (NSFs) are candidates for resolving these problems. This paper describes the fabrication of an NSF on a flexible substrate, which can be used for room temperature conductive bonding. The template method is used for preparing high-density nanowire arrays. A Cu thin film is layered on the template as the flexible substrate. After etching the template, a Cu NSF is obtained on the Cu film substrate. In addition, the electrical and mechanical properties of the Cu NSF are studied under various fabrication conditions. The Cu NSF exhibits high shear adhesion strength (∼234 N cm‑2) and low contact resistivity (2.2 × 10‑4 Ω cm2).

  13. Yttrium aluminum garnet coating on glass substrate

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Camila M.A.; Freiria, Gabriela S.; Faria, Emerson H. de; Rocha, Lucas A.; Ciuffi, Katia J.; Nassar, Eduardo J., E-mail: eduardo.nassar@unifran.edu.br

    2016-02-15

    Thin luminescent films have seen great technological advances and are applicable in the production of a variety of materials such as sensors, solar cells, photovoltaic devices, optical magnetic readers, waveguides, lasers, and recorders. Systems that contain yttrium aluminum oxide are important hosts for lanthanide ions and serve as light emission devices. This work deals with the deposition of yttrium aluminum garnet (YAG) film doped with Eu{sup 3+} onto a glass substrate obtained by the sol–gel methodology. Spray pyrolysis furnished the yttrium aluminum oxide powder. Dip-coating at a withdrawal speed of 10 mm min{sup −1} and evaporation led to deposition of different numbers of layers of the YAG:Eu{sup 3+} film onto the glass substrate from a YAG:Eu{sup 3+} powder suspension containing ethanol, water, and tetraethylorthosilicate. Photoluminescence, X-ray diffraction, scanning electron microscopy, and transparency measurements aided film characterization. The emission spectra revealed that the number of layers influenced film properties. - Highlights: • The spray pyrolysis was used to obtain luminescent YAG:Eu{sup 3+}. • The matrix was deposited as transparent films. • The YAG:Eu{sup 3+} was deposited by sol–gel process onto glass substrate.

  14. Yttrium aluminum garnet coating on glass substrate

    International Nuclear Information System (INIS)

    Ferreira, Camila M.A.; Freiria, Gabriela S.; Faria, Emerson H. de; Rocha, Lucas A.; Ciuffi, Katia J.; Nassar, Eduardo J.

    2016-01-01

    Thin luminescent films have seen great technological advances and are applicable in the production of a variety of materials such as sensors, solar cells, photovoltaic devices, optical magnetic readers, waveguides, lasers, and recorders. Systems that contain yttrium aluminum oxide are important hosts for lanthanide ions and serve as light emission devices. This work deals with the deposition of yttrium aluminum garnet (YAG) film doped with Eu 3+ onto a glass substrate obtained by the sol–gel methodology. Spray pyrolysis furnished the yttrium aluminum oxide powder. Dip-coating at a withdrawal speed of 10 mm min −1 and evaporation led to deposition of different numbers of layers of the YAG:Eu 3+ film onto the glass substrate from a YAG:Eu 3+ powder suspension containing ethanol, water, and tetraethylorthosilicate. Photoluminescence, X-ray diffraction, scanning electron microscopy, and transparency measurements aided film characterization. The emission spectra revealed that the number of layers influenced film properties. - Highlights: • The spray pyrolysis was used to obtain luminescent YAG:Eu 3+ . • The matrix was deposited as transparent films. • The YAG:Eu 3+ was deposited by sol–gel process onto glass substrate.

  15. Quantitative framework for ordered degradation of APC/C substrates.

    Science.gov (United States)

    Lu, Dan; Girard, Juliet R; Li, Weihan; Mizrak, Arda; Morgan, David O

    2015-11-16

    During cell-cycle progression, substrates of a single master regulatory enzyme can be modified in a specific order. Here, we used experimental and computational approaches to dissect the quantitative mechanisms underlying the ordered degradation of the substrates of the ubiquitin ligase APC/C(Cdc20), a key regulator of chromosome segregation in mitosis. We show experimentally that the rate of catalysis varies with different substrates of APC/C(Cdc20). Using a computational model based on multi-step ubiquitination, we then show how changes in the interaction between a single substrate and APC/C(Cdc20) can alter the timing of degradation onset relative to APC/C(Cdc20) activation, while ensuring a fast degradation rate. Degradation timing and dynamics depend on substrate affinity for the enzyme as well as the catalytic rate at which the substrate is modified. When two substrates share the same pool of APC/C(Cdc20), their relative enzyme affinities and rates of catalysis influence the partitioning of APC/C(Cdc20) among substrates, resulting in substrate competition. Depending on how APC/C(Cdc20) is partitioned among its substrates, competition can have minor or major effects on the degradation of certain substrates. We show experimentally that increased expression of the early APC/C(Cdc20) substrate Clb5 does not delay the degradation of the later substrate securin, arguing against a role for competition with Clb5 in establishing securin degradation timing. The degradation timing of APC/C(Cdc20) substrates depends on the multi-step nature of ubiquitination, differences in substrate-APC/C(Cdc20) interactions, and competition among substrates. Our studies provide a conceptual framework for understanding how ordered modification can be established among substrates of the same regulatory enzyme, and facilitate our understanding of how precise temporal control is achieved by a small number of master regulators to ensure a successful cell division cycle.

  16. Surface control alloy substrates and methods of manufacture therefor

    Energy Technology Data Exchange (ETDEWEB)

    Fritzemeier, Leslie G. (Mendon, MA); Li, Qi (Marlborough, MA); Rupich, Martin W. (Framingham, MA); Thompson, Elliott D. (Coventry, RI); Siegal, Edward J. (Malden, MA); Thieme, Cornelis Leo Hans (Westborough, MA); Annavarapu, Suresh (Brookline, MA); Arendt, Paul N. (Los Alamos, NM); Foltyn, Stephen R. (Los Alamos, NM)

    2004-05-04

    Methods and articles for controlling the surface of an alloy substrate for deposition of an epitaxial layer. The invention includes the use of an intermediate layer to stabilize the substrate surface against oxidation for subsequent deposition of an epitaxial layer.

  17. Substrate-induced instability in gas microstrip detectors

    International Nuclear Information System (INIS)

    Bateman, J.E.; Connolly, J.F.

    1992-12-01

    The results of a programme of research into substrate-induced gain instability in gas microstrip detectors are reported. Information has been collected on a wide range of substrates including many commonly available glasses and ceramics. A theoretical model of the gain instability is proposed. While we have not yet found an acceptable substrate for the construction of high flux detectors our experience points to electronically conductive glasses as the most promising source of a stable substrate. (Author)

  18. Analysis of Rectangular Microstrip Antennas with Air Substrates ...

    African Journals Online (AJOL)

    This paper presents an analysis of rectangular microstrip antennas with air substrates. The effect of the substrate thickness on the bandwidth and the efficiency are examined. An additional thin layer supporting the dielectric material is added to the air substrate in order to make the antenna mechanically rigid and easy to ...

  19. Effect of Substrate on Biogas Yield | Adamu | Global Journal of ...

    African Journals Online (AJOL)

    Biogas technology converts biological matter or biological waste (substrate) into energy and simultaneously helps to improve the quality of life and the environment. The effect of substrate on biogas yield was studied by using different substrate in laboratory scale experiment using water displacement method to monitor the ...

  20. Graphene-on-semiconductor substrates for analog electronics

    Science.gov (United States)

    Lagally, Max G.; Cavallo, Francesca; Rojas-Delgado, Richard

    2016-04-26

    Electrically conductive material structures, analog electronic devices incorporating the structures and methods for making the structures are provided. The structures include a layer of graphene on a semiconductor substrate. The graphene layer and the substrate are separated by an interfacial region that promotes transfer of charge carriers from the surface of the substrate to the graphene.

  1. Toward robust nanogenerators using aluminum substrate

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sangmin; Xu, Chen; Lee, Minbaek; Lin, Long [School of Material Science and Engineering, Georgia Institute of Technology, Atlanta, Georgia (United States); Hong, Jung-Il [School of Material Science and Engineering, Georgia Institute of Technology, Atlanta, Georgia (United States); Department of Emerging Materials Science, Daegu Gyeongbuk Institute of Science and Technology, Daegu (Korea, Republic of); Kim, Dongseob; Hwang, Woonbong [Department of Mechanical Engineering, Pohang University of Science and Technology, San 31, Hyoja, Namgu, Pohang, Gyungbuk (Korea, Republic of); Wang, Zhong Lin [School of Material Science and Engineering, Georgia Institute of Technology, Atlanta, Georgia (United States); Beijing Institue of Nanoenergy and Nanosystems, Chinese Academy of Sciences, Beijing (China)

    2012-08-22

    Nanogenerators (NG) have been developed to harvest mechanical energy from environmental sources such as vibration, human motion, or movement of automobiles. We demonstrate a robust and large-area NG based on a cost-effective Al substrate with the capability to be easily integrated in series and parallel for high-output performance. The output voltage and current density of the three-dimensionally integrated NG device reaches up to 3 V and 195 nA under human walking conditions. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  2. Metallic substrates for high temperature superconductors

    Science.gov (United States)

    Truchan, Thomas G.; Miller, Dean J.; Goretta, Kenneth C.; Balachandran, Uthamalingam; Foley, Robert

    2002-01-01

    A biaxially textured face-centered cubic metal article having grain boundaries with misorientation angles greater than about 8.degree. limited to less than about 1%. A laminate article is also disclosed having a metal substrate first rolled to at least about 95% thickness reduction followed by a first annealing at a temperature less than about 375.degree. C. Then a second rolling operation of not greater than about 6% thickness reduction is provided, followed by a second annealing at a temperature greater than about 400.degree. C. A method of forming the metal and laminate articles is also disclosed.

  3. Ferromagnetic Film on a Superconducting Substrate

    OpenAIRE

    Bulaevskii, L. N.; Chudnovsky, E. M.

    2000-01-01

    We study the equilibrium domain structure and magnetic flux around a ferromagnetic (FM) film with perpendicular magnetization M_0 on a superconducting (SC) substrate. At 4{\\pi}M_0> 1. Here \\lambda_L is the London penetration length. For 4{\\pi}M_0 > H_{c1} and l_{N} in excess of about 35 {\\lambda}_{L}, the domains are connected by SC vortices. We argue that pinning of vortices by magnetic domains in FM/SC multilayers can provide high critical currents.

  4. Molybdenum protective coatings adhesion to steel substrate

    Science.gov (United States)

    Blesman, A. I.; Postnikov, D. V.; Polonyankin, D. A.; Teplouhov, A. A.; Tyukin, A. V.; Tkachenko, E. A.

    2017-06-01

    Protection of the critical parts, components and assemblies from corrosion is an urgent engineering problem and many other industries. Protective coatings’ forming on surface of metal products is a promising way of corrosionprevention. The adhesion force is one of the main characteristics of coatings’ durability. The paper presents theoretical and experimental adhesion force assessment for coatings formed by molybdenum magnetron sputtering ontoa steel substrate. Validity and reliability of results obtained by simulation and sclerometry method allow applying the developed model for adhesion force evaluation in binary «steel-coating» systems.

  5. Ventricular tachycardia in ischemic heart disease substrates

    Directory of Open Access Journals (Sweden)

    Olujimi A. Ajijola

    2014-01-01

    This review will discuss the central role of the ischemic heart disease substrate in the development MMVT. Electrophysiologic characterization of the post-infarct myocardium using bipolar electrogram amplitudes to delineate scar border zones will be reviewed. Functional electrogram determinants of reentrant circuits such as isolated late potentials will be discussed. Strategies for catheter ablation of reentrant ventricular tachycardia, including structural and functional targets will also be examined, as will the role of the epicardial mapping and ablation in the management of recurrent MMVT.

  6. Macro algae as substrate for biogas production

    DEFF Research Database (Denmark)

    Møller, Henrik; Sarker, Shiplu; Gautam, Dhan Prasad

    Algae as a substrate for biogas is superior to other crops since it has a much higher yield of biomass per unit area and since algae grows in the seawater there will be no competition with food production on agricultural lands. So far, the progress in treating different groups of algae as a source...... of energy is promising. In this study 5 different algae types were tested for biogas potential and two algae were subsequent used for co-digestion with manure. Green seaweed, Ulva lactuca and brown seaweed Laminaria digitata was co-digested with cattle manure at mesophilic and thermophilic condition...

  7. Gas microstrip detectors on resistive plastic substrates

    International Nuclear Information System (INIS)

    Dixit, M.S.; Oakham, F.G.; Armitage, J.C.

    1993-01-01

    Plastics are desirable as substrates for gas microstrip detectors (GMDs) because of their flexibility, low density and long radiation length. GMDs have been fabricated on white Tedlar which has bulk electrical conductivity and ion-implanted Upilex which has a thin electrically conductive layer on the surface of an insulator. The effect of back plane voltage on the gain of such GMDs is investigated. Three 200 μm pitch, ion-implanted Upilex GMDs were recently tested in a high intensity beam at CERN. The anode signals were read out using fast, low noise, high gain amplifiers. Preliminary results of the test are presented

  8. Protective amorphous carbon coatings on glass substrates

    Science.gov (United States)

    Silins, Kaspars; Baránková, Hana; Bardos, Ladislav

    2017-11-01

    Thick amorphous carbon films were deposited by the Magnets-in-Motion (M-M) rf linear hollow cathode at varying acetylene contents in Ar in a hybrid PVD/PE-CVD process directly on glass substrates. The hollow cathode plates manufactured from graphite were used as the PVD target. The measurements show that the films can reach thickness of up to 50 μm at deposition rates of up to 2.5 μm/min. Scratch test measurements confirm that well adhering films several μm thick can be achieved at C2H2 contents of up to 0.5%.

  9. Method of forming through substrate vias (TSVs) and singulating and releasing die having the TSVs from a mechanical support substrate

    Science.gov (United States)

    Okandan, Murat; Nielson, Gregory N

    2014-12-09

    Accessing a workpiece object in semiconductor processing is disclosed. The workpiece object includes a mechanical support substrate, a release layer over the mechanical support substrate, and an integrated circuit substrate coupled over the release layer. The integrated circuit substrate includes a device layer having semiconductor devices. The method also includes etching through-substrate via (TSV) openings through the integrated circuit substrate that have buried ends at or within the release layer including using the release layer as an etch stop. TSVs are formed by introducing one or more conductive materials into the TSV openings. A die singulation trench is etched at least substantially through the integrated circuit substrate around a perimeter of an integrated circuit die. The integrated circuit die is at least substantially released from the mechanical support substrate.

  10. Neural substrates of sublexical processing for spelling.

    Science.gov (United States)

    DeMarco, Andrew T; Wilson, Stephen M; Rising, Kindle; Rapcsak, Steven Z; Beeson, Pélagie M

    2017-01-01

    We used fMRI to examine the neural substrates of sublexical phoneme-grapheme conversion during spelling in a group of healthy young adults. Participants performed a writing-to-dictation task involving irregular words (e.g., choir), plausible nonwords (e.g., kroid), and a control task of drawing familiar geometric shapes (e.g., squares). Written production of both irregular words and nonwords engaged a left-hemisphere perisylvian network associated with reading/spelling and phonological processing skills. Effects of lexicality, manifested by increased activation during nonword relative to irregular word spelling, were noted in anterior perisylvian regions (posterior inferior frontal gyrus/operculum/precentral gyrus/insula), and in left ventral occipito-temporal cortex. In addition to enhanced neural responses within domain-specific components of the language network, the increased cognitive demands associated with spelling nonwords engaged domain-general frontoparietal cortical networks involved in selective attention and executive control. These results elucidate the neural substrates of sublexical processing during written language production and complement lesion-deficit correlation studies of phonological agraphia. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Alloyed surfaces: New substrates for graphene growth

    Science.gov (United States)

    Tresca, C.; Verbitskiy, N. I.; Fedorov, A.; Grüneis, A.; Profeta, G.

    2017-11-01

    We report a systematic ab-initio density functional theory investigation of Ni(111) surface alloyed with elements of group IV (Si, Ge and Sn), demonstrating the possibility to use it to grow high quality graphene. Ni(111) surface represents an ideal substrate for graphene, due to its catalytic properties and perfect matching with the graphene lattice constant. However, Dirac bands of graphene growth on Ni(111) are completely destroyed due to the strong hybridization between carbon pz and Ni d orbitals. Group IV atoms, namely Si, Ge and Sn, once deposited on Ni(111) surface, form an ordered alloyed surface with √{ 3} ×√{ 3} -R30° reconstruction. We demonstrate that, at variance with the pure Ni(111) surface, alloyed surfaces effectively decouple graphene from the substrate, resulting unstrained due to the nearly perfect lattice matching and preserves linear Dirac bands without the strong hybridization with Ni d states. The proposed surfaces can be prepared before graphene growth without resorting on post-growth processes which necessarily alter the electronic and structural properties of graphene.

  12. Memory Consolidation and Neural Substrate of Reward

    Directory of Open Access Journals (Sweden)

    Redolar-Ripoll, Diego

    2012-08-01

    Full Text Available The aim of this report is to analyze the relationships between reward and learning and memory processes. Different studies have described how information about rewards influences behavior and how the brain uses this reward information to control learning and memory processes. Reward nature seems to be processed in different ways by neurons in different brain structures, ranging from the detection and perception of rewards to the use of information about predicted rewards for the control of goal-directed behavior. The neural substrate underling this processing of reward information is a reliable way of improving learning and memory processes. Evidence from several studies indicates that this neural system can facilitate memory consolidation in a wide variety of learning tasks. From a molecular perspective, certain cardinal features of reward have been described as forms of memory. Studies of human addicts and studies in animal models of addiction show that chronic drug exposure produces stable changes in the brain at the cellular and molecular levels that underlie the long-lasting behavioral plasticity associated with addiction. These molecular and cellular adaptations involved in addiction are also implicated in learning and memory processes. Dopamine seems to be a critical common signal to activate different genetic mechanisms that ultimately remodel synapses and circuits. Despite memory is an active and complex process mediated by different brain areas, the neural substrate of reward is able to improve memory consolidation in a several paradigms. We believe that there are many equivalent traits between reward and learning and memory processes.

  13. Electrospinning onto Insulating Substrates by Controlling Surface Wettability and Humidity

    Science.gov (United States)

    Choi, WooSeok; Kim, Geon Hwee; Shin, Jung Hwal; Lim, Geunbae; An, Taechang

    2017-11-01

    We report a simple method for electrospinning polymers onto flexible, insulating substrates by controlling the wettability of the substrate surface. Water molecules were adsorbed onto the surface of a hydrophilic polymer substrate by increasing the local humidity around the substrate. The adsorbed water was used as the ground electrode for electrospinning. The electrospun fibers were deposited only onto hydrophilic areas of the substrate, allowing for patterning through wettability control. Direct writing of polymer fiber was also possible through near-field electrospinning onto a hydrophilic surface.

  14. SOI MESFETs on high-resistivity, trap-rich substrates

    Science.gov (United States)

    Mehr, Payam; Zhang, Xiong; Lepkowski, William; Li, Chaojiang; Thornton, Trevor J.

    2018-04-01

    The DC and RF characteristics of metal-semiconductor field-effect-transistors (MESFETs) on conventional CMOS silicon-on-insulator (SOI) substrates are compared to nominally identical devices on high-resistivity, trap-rich SOI substrates. While the DC transfer characteristics are statistically identical on either substrate, the maximum available gain at GHz frequencies is enhanced by ∼2 dB when using the trap-rich substrates, with maximum operating frequencies, fmax, that are approximately 5-10% higher. The increased fmax is explained by the reduced substrate conduction at GHz frequencies using a lumped-element, small-signal model.

  15. Nemesia root hair response to paper pulp substrate for micropropagation.

    Science.gov (United States)

    Labrousse, Pascal; Delmail, David; Decou, Raphaël; Carlué, Michel; Lhernould, Sabine; Krausz, Pierre

    2012-01-01

    Agar substrates for in vitro culture are well adapted to plant micropropagation, but not to plant rooting and acclimatization. Conversely, paper-pulp-based substrates appear as potentially well adapted for in vitro culture and functional root production. To reinforce this hypothesis, this study compares in vitro development of nemesia on several substrates. Strong differences between nemesia roots growing in agar or in paper-pulp substrates were evidenced through scanning electron microscopy. Roots developed in agar have shorter hairs, larger rhizodermal cells, and less organized root caps than those growing on paper pulp. In conclusion, it should be noted that in this study, in vitro microporous substrates such as paper pulp lead to the production of similar root hairs to those found in greenhouse peat substrates. Consequently, if agar could be used for micropropagation, rooting, and plant acclimatization, enhancement could be achieved if rooting stage was performed on micro-porous substrates such as paper pulp.

  16. Chemometrics approach to substrate development, case: semisyntetic cheese

    DEFF Research Database (Denmark)

    Nielsen, Per Væggemose; Hansen, Birgitte Vedel

    1998-01-01

    from food production facilities.The Chemometrics approach to substrate development is illustrated by the development of a semisyntetic cheese substrate. Growth, colour formation and mycotoxin production of 6 cheese related fungi were studied on 9 types of natural cheeses and 24 synthetic cheese......, the most frequently occurring contaminant on semi-hard cheese. Growth experiments on the substrate were repeatable and reproducible. The substrate was also suitable for the starter P. camemberti. Mineral elements in cheese were shown to have strong effect on growth, mycotoxin production and colour...... formation of fungi. For P. roqueforti, P. discolor, P. verrucosum and Aspergillus versicolor the substrate was less suitable as a model cheese substrate, which indicates great variation in nutritional demands of the fungi. Substrates suitable for studies of specific cheese types was found for P. roqueforti...

  17. An Analytical Model for Spectral Peak Frequency Prediction of Substrate Noise in CMOS Substrates

    DEFF Research Database (Denmark)

    Shen, Ming; Mikkelsen, Jan H.

    2013-01-01

    This paper proposes an analytical model describing the generation of switching current noise in CMOS substrates. The model eliminates the need for SPICE simulations in existing methods by conducting a transient analysis on a generic CMOS inverter and approximating the switching current waveform us...

  18. Automatic classification of blank substrate defects

    Science.gov (United States)

    Boettiger, Tom; Buck, Peter; Paninjath, Sankaranarayanan; Pereira, Mark; Ronald, Rob; Rost, Dan; Samir, Bhamidipati

    2014-10-01

    Mask preparation stages are crucial in mask manufacturing, since this mask is to later act as a template for considerable number of dies on wafer. Defects on the initial blank substrate, and subsequent cleaned and coated substrates, can have a profound impact on the usability of the finished mask. This emphasizes the need for early and accurate identification of blank substrate defects and the risk they pose to the patterned reticle. While Automatic Defect Classification (ADC) is a well-developed technology for inspection and analysis of defects on patterned wafers and masks in the semiconductors industry, ADC for mask blanks is still in the early stages of adoption and development. Calibre ADC is a powerful analysis tool for fast, accurate, consistent and automatic classification of defects on mask blanks. Accurate, automated classification of mask blanks leads to better usability of blanks by enabling defect avoidance technologies during mask writing. Detailed information on blank defects can help to select appropriate job-decks to be written on the mask by defect avoidance tools [1][4][5]. Smart algorithms separate critical defects from the potentially large number of non-critical defects or false defects detected at various stages during mask blank preparation. Mechanisms used by Calibre ADC to identify and characterize defects include defect location and size, signal polarity (dark, bright) in both transmitted and reflected review images, distinguishing defect signals from background noise in defect images. The Calibre ADC engine then uses a decision tree to translate this information into a defect classification code. Using this automated process improves classification accuracy, repeatability and speed, while avoiding the subjectivity of human judgment compared to the alternative of manual defect classification by trained personnel [2]. This paper focuses on the results from the evaluation of Automatic Defect Classification (ADC) product at MP Mask

  19. Plastic properties of thin films on substrates as measured by submicron indentation hardness and substrate curvature techniques

    International Nuclear Information System (INIS)

    Doerner, M.F.; Gardner, D.S.; Nix, W.D.

    1986-01-01

    Substrate curvature and submicron indentation measurements have been used recently to study plastic deformation in thin films on substrates. In the present work both of these techniques have been employed to study the strength of aluminum and tungsten thin films on silicon substrates. In the case of aluminum films on silicon substrates, the film strength is found to increase with decreasing thickness. Grain size variations with film thickness do not account for the variations in strength. Wafer curvature measurements give strengths higher than those predicted from hardness measurements suggesting the substrate plays a role in strengthening the film. The observed strengthening effect with decreased thickness may be due to image forces on dislocations in the film due to the elastically stiffer silicon substrate. For sputtered tungsten films, where the substrate is less stiff than the film, the film strength decreases with decreasing film thickness

  20. Vector assembly of colloids on monolayer substrates

    Science.gov (United States)

    Jiang, Lingxiang; Yang, Shenyu; Tsang, Boyce; Tu, Mei; Granick, Steve

    2017-06-01

    The key to spontaneous and directed assembly is to encode the desired assembly information to building blocks in a programmable and efficient way. In computer graphics, raster graphics encodes images on a single-pixel level, conferring fine details at the expense of large file sizes, whereas vector graphics encrypts shape information into vectors that allow small file sizes and operational transformations. Here, we adapt this raster/vector concept to a 2D colloidal system and realize `vector assembly' by manipulating particles on a colloidal monolayer substrate with optical tweezers. In contrast to raster assembly that assigns optical tweezers to each particle, vector assembly requires a minimal number of optical tweezers that allow operations like chain elongation and shortening. This vector approach enables simple uniform particles to form a vast collection of colloidal arenes and colloidenes, the spontaneous dissociation of which is achieved with precision and stage-by-stage complexity by simply removing the optical tweezers.

  1. A universal mammalian vaccine cell line substrate.

    Directory of Open Access Journals (Sweden)

    Jackelyn Murray

    Full Text Available Using genome-wide small interfering RNA (siRNA screens for poliovirus, influenza A virus and rotavirus, we validated the top 6 gene hits PV, RV or IAV to search for host genes that when knocked-down (KD enhanced virus permissiveness and replication over wild type Vero cells or HEp-2 cells. The enhanced virus replication was tested for 12 viruses and ranged from 2-fold to >1000-fold. There were variations in virus-specific replication (strain differences across the cell lines examined. Some host genes (CNTD2, COQ9, GCGR, NDUFA9, NEU2, PYCR1, SEC16G, SVOPL, ZFYVE9, and ZNF205 showed that KD resulted in enhanced virus replication. These findings advance platform-enabling vaccine technology, the creation of diagnostic cells substrates, and are informative about the host mechanisms that affect virus replication in mammalian cells.

  2. Graphene based strain sensor with LCP substrate

    Science.gov (United States)

    Nie, M.; Yang, H. S.; Xia, Y. H.

    2018-02-01

    A flexible strain sensor constructed by an efficient, low-cost fabrication strategy is presented in this paper. It is assembled by adhering grid-like graphene on LCP substrate. Kinds of measurement setup have been designed to verify that the proposed flexible sensor device is suitable to be used in health monitoring system. From the experiment results, it can be proved that the sensor exhibits the following features: ultra-light, relatively good sensitivity, high reversibility, superior physical robustness, easy fabrication. With the great performance of this flexible strain sensor, it is considered to play an important role in body monitoring, structural health monitoring system, fatigue detection and healthcare systems in the near future.

  3. Deposition of silver layer on different substrates

    Science.gov (United States)

    Krzemiński, J.; Kiełbasiński, K.; Szałapak, J.; Jakubowska, M.; MłoŻniak, A.; Zwierkowska, E.

    2015-09-01

    The hole process of producing continuous layer with silver nanoparticles is presented in this paper. First the ink preparation and then the spray process is shown and discussed. The silver layers were obtained on sodium glass substrate. Three different ink carriers were considered and the best one has been chosen. Spray coating process was carried out using special spray can. After obtaining sprayed layers the samples were sintered in several temperatures to investigate the lowest suitable sintering temperature. After that layers resistivity were measured. Then the silver layers were cracked to produce breakthrough fracture that was investigated by a scanning electron microscope. In this paper, the authors investigated the spray coating technique as an alternative to electroplating and other techniques, considering layer resistivity, thickness and production process.

  4. The structural neural substrate of subjective happiness.

    Science.gov (United States)

    Sato, Wataru; Kochiyama, Takanori; Uono, Shota; Kubota, Yasutaka; Sawada, Reiko; Yoshimura, Sayaka; Toichi, Motomi

    2015-11-20

    Happiness is a subjective experience that is an ultimate goal for humans. Psychological studies have shown that subjective happiness can be measured reliably and consists of emotional and cognitive components. However, the neural substrates of subjective happiness remain unclear. To investigate this issue, we used structural magnetic resonance imaging and questionnaires that assessed subjective happiness, the intensity of positive and negative emotional experiences, and purpose in life. We found a positive relationship between the subjective happiness score and gray matter volume in the right precuneus. Moreover, the same region showed an association with the combined positive and negative emotional intensity and purpose in life scores. Our findings suggest that the precuneus mediates subjective happiness by integrating the emotional and cognitive components of happiness.

  5. Micropatterned substrates for studying astrocytes in culture

    Directory of Open Access Journals (Sweden)

    William Lee

    2009-12-01

    Full Text Available Recent studies of the physiological roles of astrocytes have ignited renewed interest in the functional significance of these glial cells in the central nervous system. Many of the newly discovered astrocytic functions were initially demonstrated and characterized in cell culture systems. We discuss the use of microculture techniques and micropatterning of cell-adhesive substrates in studies of astrocytic Ca2+ excitability and bidirectional neuron-astrocyte signaling. This culturing approach aims to reduce the level of complexity of the system by limiting the interacting partners and by controlling the localization of cells. It provides tight control over experimental conditions allowing detailed characterization of cellular functions and intercellular communication. Although such a reductionist approach yields some difference in observations between astrocytic properties in culture and in situ, general phenomena discovered in cell culture systems, however, have also been found in vivo.

  6. Experimental analysis of green roof substrate detention characteristics.

    Science.gov (United States)

    Yio, Marcus H N; Stovin, Virginia; Werdin, Jörg; Vesuviano, Gianni

    2013-01-01

    Green roofs may make an important contribution to urban stormwater management. Rainfall-runoff models are required to evaluate green roof responses to specific rainfall inputs. The roof's hydrological response is a function of its configuration, with the substrate - or growing media - providing both retention and detention of rainfall. The objective of the research described here is to quantify the detention effects due to green roof substrates, and to propose a suitable hydrological modelling approach. Laboratory results from experimental detention tests on green roof substrates are presented. It is shown that detention increases with substrate depth and as a result of increasing substrate organic content. Model structures based on reservoir routing are evaluated, and it is found that a one-parameter reservoir routing model coupled with a parameter that describes the delay to start of runoff best fits the observed data. Preliminary findings support the hypothesis that the reservoir routing parameter values can be defined from the substrate's physical characteristics.

  7. Substrate structures for InP-based devices

    International Nuclear Information System (INIS)

    Wanlass, M.W.; Sheldon, P.

    1990-01-01

    A substrate structure for an InP-based semiconductor device having an InP based film is described. The substrate structure includes a substrate region having a lightweight bulk substrate and an upper GaAs layer. An interconnecting region is disposed between the substrate region and the InP-based device. The interconnecting region includes a compositionally graded intermediate layer substantially lattice-matched at the opposite end to the InP=based film. The interconnecting region further includes a dislocation mechanism disposed between the GaAs layer and the InP-based film in cooperation with the graded intermediate layer, the buffer mechanism blocking and inhibiting propagation of threading dislocations between the substrate region, and the InP-based device

  8. Microwave GaAs Integrated Circuits On Quartz Substrates

    Science.gov (United States)

    Siegel, Peter H.; Mehdi, Imran; Wilson, Barbara

    1994-01-01

    Integrated circuits for use in detecting electromagnetic radiation at millimeter and submillimeter wavelengths constructed by bonding GaAs-based integrated circuits onto quartz-substrate-based stripline circuits. Approach offers combined advantages of high-speed semiconductor active devices made only on epitaxially deposited GaAs substrates with low-dielectric-loss, mechanically rugged quartz substrates. Other potential applications include integration of antenna elements with active devices, using carrier substrates other than quartz to meet particular requirements using lifted-off GaAs layer in membrane configuration with quartz substrate supporting edges only, and using lift-off technique to fabricate ultrathin discrete devices diced separately and inserted into predefined larger circuits. In different device concept, quartz substrate utilized as transparent support for GaAs devices excited from back side by optical radiation.

  9. Vitreous carbon mask substrate for X-ray lithography

    Science.gov (United States)

    Aigeldinger, Georg [Livermore, CA; Skala, Dawn M [Fremont, CA; Griffiths, Stewart K [Livermore, CA; Talin, Albert Alec [Livermore, CA; Losey, Matthew W [Livermore, CA; Yang, Chu-Yeu Peter [Dublin, CA

    2009-10-27

    The present invention is directed to the use of vitreous carbon as a substrate material for providing masks for X-ray lithography. The new substrate also enables a small thickness of the mask absorber used to pattern the resist, and this enables improved mask accuracy. An alternative embodiment comprised the use of vitreous carbon as a LIGA substrate wherein the VC wafer blank is etched in a reactive ion plasma after which an X-ray resist is bonded. This surface treatment provides a surface enabling good adhesion of the X-ray photoresist and subsequent nucleation and adhesion of the electrodeposited metal for LIGA mold-making while the VC substrate practically eliminates secondary radiation effects that lead to delamination of the X-ray resist form the substrate, the loss of isolated resist features, and the formation of a resist layer adjacent to the substrate that is insoluble in the developer.

  10. Pd thin films on flexible substrate for hydrogen sensor

    Energy Technology Data Exchange (ETDEWEB)

    Öztürk, Sadullah [Fatih Sultan Mehmet Vakıf University, Engineering Faculty, Istanbul (Turkey); Kılınç, Necmettin, E-mail: nkilinc@nigde.edu.tr [Nigde University, Mechatronics Engineering Department, 51245 Nigde (Turkey); Nigde University, Nanotechnology Application and Research Center, 51245 Nigde (Turkey)

    2016-07-25

    In this work, palladium (Pd) thin films were prepared via RF sputtering method with various thicknesses (6 nm, 20 nm and 60 nm) on both a flexible substrate and a hard substrate. Hydrogen (H{sub 2}) sensing properties of Pd films on flexible substrate have been investigated depending on temperatures (25–100 °C) and H{sub 2} concentrations (600 ppm – 10%). The effect of H{sub 2} on structural properties of the films was also studied. The films were characterized by Scanning Electron Microscopy (SEM) and X-ray diffraction. It is found that whole Pd films on hard substrate show permanent structural deformation after exposed to 10% H{sub 2} for 30 min. But, this H{sub 2} exposure does not causes any structural deformation for 6 nm Pd film on flexible substrate and 6 nm Pd film on flexible substrate shows reversible sensor response up to 10% H{sub 2} concentration without any structural deformation. On the other hand, Pd film sensors that have the thicknesses 20 nm and 60 nm on flexible substrate are irreversible for higher H{sub 2} concentration (>2%) with film deformation. The sensor response of 6 nm Pd film on flexible substrate increased with increasing H{sub 2} concentration up 4% and then saturated. The sensitivity of the film decreased with increasing operation temperature. - Highlights: • Pd thin films fabricated by RF sputtering on both flexible and hard substrates. • Structural deformation observed for films on hard substrate after exposing 10% H{sub 2}. • 6 nm Pd film on flexible substrate shows reversible sensor response up to 10% H{sub 2}. • H{sub 2} sensing properties of film on flexible substrate investigated depending on temperature and concentration. • The sensitivity of the film decreased with increasing operation temperature.

  11. Mogul-Patterned Elastomeric Substrate for Stretchable Electronics.

    Science.gov (United States)

    Lee, Han-Byeol; Bae, Chan-Wool; Duy, Le Thai; Sohn, Il-Yung; Kim, Do-Il; Song, You-Joon; Kim, Youn-Jea; Lee, Nae-Eung

    2016-04-01

    A mogul-patterned stretchable substrate with multidirectional stretchability and minimal fracture of layers under high stretching is fabricated by double photolithography and soft lithography. Au layers and a reduced graphene oxide chemiresistor on a mogul-patterned poly(dimethylsiloxane) substrate are stable and durable under various stretching conditions. The newly designed mogul-patterned stretchable substrate shows great promise for stretchable electronics. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Wetting Behavior in Colloid-Polymer Mixtures at Different Substrates.

    Science.gov (United States)

    Wijting, Willem K; Besseling, Nicolaas A M; Cohen Stuart, Martien A

    2003-09-25

    We present experimental observations on wetting phenomena in depletion interaction driven, phase separated colloidal dispersions. The contact angle of the colloidal liquid-gas interface at a solid substrate was determined for a series of compositions. Upon approach to the critical point, a transition occurs from partial to complete wetting. The interaction with the substrate was manipulated by modifying the substrate with a polymer. In that case, a transition from partial to complete drying is observed upon approach to the critical point.

  13. Layered graphene-mica substrates induce melting of DNA origami

    Science.gov (United States)

    Green, Nathaniel S.; Pham, Phi H. Q.; Crow, Daniel T.; Burke, Peter J.; Norton, Michael L.

    2018-04-01

    Monolayer graphene supported on mica substrates induce melting of cross-shaped DNA origami. This behavior can be contrasted with the case of origami on graphene on graphite, where an expansion or partially re-organized structure is observed. On mica, only well-formed structures are observed. Comparison of the morphological differences observed for these probes after adsorption on these substrates provides insights into the sensitivity of DNA based nanostructures to the properties of the graphene monolayer, as modified by its substrate.

  14. Substrate binding and specificity of rhomboid intramembrane protease revealed by substrate-peptide complex structures

    Czech Academy of Sciences Publication Activity Database

    Zoll, Sebastian; Stanchev, Stancho; Began, Jakub; Škerle, Jan; Lepšík, Martin; Peclinovská, Lucie; Majer, Pavel; Stříšovský, Kvido

    2014-01-01

    Roč. 33, č. 20 (2014), s. 2408-2421 ISSN 0261-4189 R&D Projects: GA ČR GAP305/11/1886; GA MŠk(CZ) LK11206; GA MŠk LO1302; GA ČR GBP208/12/G016 Institutional support: RVO:61388963 Keywords : intramembrane protease * rhomboid family * rhomboid protease * structure * substrate recognition Subject RIV: CE - Biochemistry Impact factor: 10.434, year: 2014

  15. Mechanical quality factor of mirror substrates for VIRGO

    CERN Document Server

    Amico, P; Carbone, L; Gammaitoni, L; Punturo, M; Travasso, F; Vocca, H

    2002-01-01

    Thermal noise in the mirror substrates is expected to be the main limit to the VIRGO sensitivity in the 50-500 Hz frequency range. The mechanical quality of the mirror substrates and the geometry of their suspension are expected to affect the noise level of the detector output. High mechanical Q have been obtained for different large fused silica substrates under VIRGO suspension conditions. Moreover, calcium fluoride substrates are shown to provide a more promising option for the design of future cryogenic, low thermal noise interferometers.

  16. Friction and Shear Strength at the Nanowire–Substrate Interfaces

    Directory of Open Access Journals (Sweden)

    Gu Yi

    2009-01-01

    Full Text Available Abstract The friction and shear strength of nanowire (NW–substrate interfaces critically influences the electrical/mechanical performance and life time of NW-based nanodevices. Yet, very few reports on this subject are available in the literature because of the experimental challenges involved and, more specifically no studies have been reported to investigate the configuration of individual NW tip in contact with a substrate. In this letter, using a new experimental method, we report the friction measurement between a NW tip and a substrate for the first time. The measurement was based on NW buckling in situ inside a scanning electron microscope. The coefficients of friction between silver NW and gold substrate and between ZnO NW and gold substrate were found to be 0.09–0.12 and 0.10–0.15, respectively. The adhesion between a NW and the substrate modified the true contact area, which affected the interfacial shear strength. Continuum mechanics calculation found that interfacial shear strengths between silver NW and gold substrate and between ZnO NW and gold substrate were 134–139 MPa and 78.9–95.3 MPa, respectively. This method can be applied to measure friction parameters of other NW–substrate systems. Our results on interfacial friction and shear strength could have implication on the AFM three-point bending tests used for nanomechanical characterisation.

  17. Influence of substrate rocks on Fe-Mn crust composition

    Science.gov (United States)

    Hein, J.R.; Morgan, C.L.

    1999-01-01

    Principal Component and other statistical analyses of chemical and mineralogical data of Fe-Mn oxyhydroxide crusts and their underlying rock substrates in the central Pacific indicate that substrate rocks do not influence crust composition. Two ridges near Johnston Atoll were dredged repetitively and up to seven substrate rock types were recovered from small areas of similar water depths. Crusts were analyzed mineralogically and chemically for 24 elements, and substrates were analyzed mineralogically and chemically for the 10 major oxides. Compositions of crusts on phosphatized substrates are distinctly different from crusts on substrates containing no phosphorite. However, that relationship only indicates that the episodes of phosphatization that mineralized the substrate rocks also mineralized the crusts that grew on them. A two-fold increase in copper contents in crusts that grew on phosphatized clastic substrate rocks, relative to crusts on other substrate rock types, is also associated with phosphatization and must have resulted from chemical reorganization during diagenesis. Phosphatized crusts show increases in Sr, Zn, Ca, Ba, Cu, Ce, V, and Mo contents and decreases in Fe, Si, and As contents relative to non-phosphatized crusts. Our statistical results support previous studies which show that crust compositions reflect predominantly direct precipitation from seawater (hydrogenetic), and to lesser extents reflect detrital input and diagenetic replacement of parts of the older crust generation by carbonate fluorapatite.

  18. Development of optimized nanogap plasmonic substrate for improved SERS enhancement

    Directory of Open Access Journals (Sweden)

    Jayakumar Perumal

    2017-05-01

    Full Text Available SERS enhancement factor (EF of planar substrates depends on the size and shape of the fine nanostructure forming a defect free, well-arranged matrix. Nano-lithographic process is considered to be the most advanced methods employed for the fabrication SERS substrates. Nanostructured plasmonic substrates with nanogap (NG pattern often results in stable, efficient and reproducible SERS enhancement. For such substrates, NG and their diagonal length (DL need to be optimized. Theoretically smaller NGs (∼30-40 nm or smaller results in higher SERS enhancement. However, fabrication of NG substrates below such limit is a challenge even for the most advanced lithography process. In this context, herein, we report the optimization of fabrication process, where higher SERS enhancement can be realized from larger NGs substrates by optimizing their DL of nanostructures between the NGs. Based on simulation we could demonstrate that, by optimizing the DL, SERS enhancement from larger NG substrate such as 60 and 80 nm could be comparable to that of smaller (40nm NG substrates. We envision that this concept will open up new regime in the nanofabrication of practically feasible NG based plasmonic substrates with higher SERS enhancement. Initial results of our experiments are in close agreement with our simulated study.

  19. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    International Nuclear Information System (INIS)

    Gartia, Manas Ranjan; Hsiao, Austin; Logan Liu, G; Sivaguru, Mayandi; Chen Yi

    2011-01-01

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  20. Friction and shear strength at the nanowire-substrate interfaces.

    Science.gov (United States)

    Zhu, Yong; Qin, Qingquan; Gu, Yi; Wang, Zhonglin

    2009-11-28

    The friction and shear strength of nanowire (NW)-substrate interfaces critically influences the electrical/mechanical performance and life time of NW-based nanodevices. Yet, very few reports on this subject are available in the literature because of the experimental challenges involved and, more specifically no studies have been reported to investigate the configuration of individual NW tip in contact with a substrate. In this letter, using a new experimental method, we report the friction measurement between a NW tip and a substrate for the first time. The measurement was based on NW buckling in situ inside a scanning electron microscope. The coefficients of friction between silver NW and gold substrate and between ZnO NW and gold substrate were found to be 0.09-0.12 and 0.10-0.15, respectively. The adhesion between a NW and the substrate modified the true contact area, which affected the interfacial shear strength. Continuum mechanics calculation found that interfacial shear strengths between silver NW and gold substrate and between ZnO NW and gold substrate were 134-139 MPa and 78.9-95.3 MPa, respectively. This method can be applied to measure friction parameters of other NW-substrate systems. Our results on interfacial friction and shear strength could have implication on the AFM three-point bending tests used for nanomechanical characterisation.

  1. Substrate Effects in Wideband SiGe HBT Mixer Circuits

    DEFF Research Database (Denmark)

    Johansen, Tom Keinicke; Vidkjær, Jens; Krozer, Viktor

    2005-01-01

    are also applied to predict short distance substrate coupling effects. Simulation results using extracted equivalent circuit models and substrate coupling networks are compared with experimental results obtained on a wideband mixer circuit implemented in a 0.35 μm, 60 GHz ft SiGe HBT BiCMOS process.......In this paper, the influence from substrate effects on the performance of wideband SiGe HBT mixer circuits is investigated. Equivalent circuit models including substrate networks are extracted from on-wafer test structures and compared with electromagnetic simulations. Electromagnetic simulations...

  2. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    Energy Technology Data Exchange (ETDEWEB)

    Gartia, Manas Ranjan [Department of Nuclear, Plasma and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Hsiao, Austin; Logan Liu, G [Department of Bioengineering, University of Illinois, Urbana, IL 61801 (United States); Sivaguru, Mayandi [Institute for Genomic Biology, University of Illinois, Urbana, IL 61801 (United States); Chen Yi, E-mail: loganliu@illinois.edu [Department of Electrical and Computer Engineering, University of Illinois, Urbana, IL 61801 (United States)

    2011-09-07

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  3. Fabrication of complex nanoscale structures on various substrates

    Science.gov (United States)

    Han, Kang-Soo; Hong, Sung-Hoon; Lee, Heon

    2007-09-01

    Polymer based complex nanoscale structures were fabricated and transferred to various substrates using reverse nanoimprint lithography. To facilitate the fabrication and transference of the large area of the nanostructured layer to the substrates, a water-soluble polyvinyl alcohol mold was used. After generation and transference of the nanostructured layer, the polyvinyl alcohol mold was removed by dissolving in water. A residue-free, UV-curable, glue layer was formulated and used to bond the nanostructured layer onto the substrates. As a result, nanometer scale patterned polymer layers were bonded to various substrates and three-dimensional nanostructures were also fabricated by stacking of the layers.

  4. Crystal Structure and Substrate Specificity of D-Galactose-6-Phosphate Isomerase Complexed with Substrates

    Science.gov (United States)

    Lee, Jung-Kul; Pan, Cheol-Ho

    2013-01-01

    D-Galactose-6-phosphate isomerase from Lactobacillus rhamnosus (LacAB; EC 5.3.1.26), which is encoded by the tagatose-6-phosphate pathway gene cluster (lacABCD), catalyzes the isomerization of D-galactose-6-phosphate to D-tagatose-6-phosphate during lactose catabolism and is used to produce rare sugars as low-calorie natural sweeteners. The crystal structures of LacAB and its complex with D-tagatose-6-phosphate revealed that LacAB is a homotetramer of LacA and LacB subunits, with a structure similar to that of ribose-5-phosphate isomerase (Rpi). Structurally, LacAB belongs to the RpiB/LacAB superfamily, having a Rossmann-like αβα sandwich fold as has been identified in pentose phosphate isomerase and hexose phosphate isomerase. In contrast to other family members, the LacB subunit also has a unique α7 helix in its C-terminus. One active site is distinctly located at the interface between LacA and LacB, whereas two active sites are present in RpiB. In the structure of the product complex, the phosphate group of D-tagatose-6-phosphate is bound to three arginine residues, including Arg-39, producing a different substrate orientation than that in RpiB, where the substrate binds at Asp-43. Due to the proximity of the Arg-134 residue and backbone Cα of the α6 helix in LacA to the last Asp-172 residue of LacB with a hydrogen bond, a six-carbon sugar-phosphate can bind in the larger pocket of LacAB, compared with RpiB. His-96 in the active site is important for ring opening and substrate orientation, and Cys-65 is essential for the isomerization activity of the enzyme. Two rare sugar substrates, D-psicose and D-ribulose, show optimal binding in the LacAB-substrate complex. These findings were supported by the results of LacA activity assays. PMID:24015281

  5. Crystal structure and substrate specificity of D-galactose-6-phosphate isomerase complexed with substrates.

    Directory of Open Access Journals (Sweden)

    Woo-Suk Jung

    Full Text Available D-Galactose-6-phosphate isomerase from Lactobacillus rhamnosus (LacAB; EC 5.3.1.26, which is encoded by the tagatose-6-phosphate pathway gene cluster (lacABCD, catalyzes the isomerization of D-galactose-6-phosphate to D-tagatose-6-phosphate during lactose catabolism and is used to produce rare sugars as low-calorie natural sweeteners. The crystal structures of LacAB and its complex with D-tagatose-6-phosphate revealed that LacAB is a homotetramer of LacA and LacB subunits, with a structure similar to that of ribose-5-phosphate isomerase (Rpi. Structurally, LacAB belongs to the RpiB/LacAB superfamily, having a Rossmann-like αβα sandwich fold as has been identified in pentose phosphate isomerase and hexose phosphate isomerase. In contrast to other family members, the LacB subunit also has a unique α7 helix in its C-terminus. One active site is distinctly located at the interface between LacA and LacB, whereas two active sites are present in RpiB. In the structure of the product complex, the phosphate group of D-tagatose-6-phosphate is bound to three arginine residues, including Arg-39, producing a different substrate orientation than that in RpiB, where the substrate binds at Asp-43. Due to the proximity of the Arg-134 residue and backbone Cα of the α6 helix in LacA to the last Asp-172 residue of LacB with a hydrogen bond, a six-carbon sugar-phosphate can bind in the larger pocket of LacAB, compared with RpiB. His-96 in the active site is important for ring opening and substrate orientation, and Cys-65 is essential for the isomerization activity of the enzyme. Two rare sugar substrates, D-psicose and D-ribulose, show optimal binding in the LacAB-substrate complex. These findings were supported by the results of LacA activity assays.

  6. Impact of substrate on soilless tomato cultivation

    Directory of Open Access Journals (Sweden)

    TP Suvo

    2016-12-01

    Full Text Available An experiment was carried out to determine the effect of four different media based hydroponics on plant growth, yield and nutritional values at Biochemistry laboratory of Patuakhali Science and Technology University (PSTU, Bangladesh during November 2014 to April 2015. Tomato plants were grown in closed soilless system where Hoagland solution as nutrient solution and jute fiber, cotton (jhut, coconut husk as substrate. Among four types of media, the media composed with Hoagland solution and jute fiber showed good impact on growth and nutritional values than the other three media (media of Hoagland solution with coconut husk, Hoagland solution with cotton and only Hoagland solution. It was revealed that the highest plant height, yield, vitamin C, fruit protein, fat and fiber content of all were related to media combination of jute fiber and Hoagland solution. Among all the verities, the highest plant height (106 cm, yield (5.3 kg plant-1, fruit Vitamin C content (64.54 mg 100 g-1, fruit protein (17.67 %, fat (5.2% and fiber (7.9% content was recorded from Patharkuchi tomato variety.

  7. Interactions of Cannabinoids With Biochemical Substrates

    Directory of Open Access Journals (Sweden)

    Brian F Thomas

    2017-05-01

    Full Text Available Recent decades have seen much progress in the identification and characterization of cannabinoid receptors and the elucidation of the mechanisms by which derivatives of the Cannabis sativa plant bind to receptors and produce their physiological and psychological effects. The information generated in this process has enabled better understanding of the fundamental physiological and psychological processes controlled by the central and peripheral nervous systems and has fostered the development of natural and synthetic cannabinoids as therapeutic agents. A negative aspect of this decades-long effort is the proliferation of clandestinely synthesized analogs as recreational street drugs with dangerous effects. Currently, the interactions of cannabinoids with their biochemical substrates are extensively but inadequately understood, and the clinical application of derived and synthetic receptor ligands remains quite limited. The wide anatomical distribution and functional complexity of the cannabinoid system continue to indicate potential for both therapeutic and side effects, which offers challenges and opportunities for medicinal chemists involved in drug discovery and development.

  8. Regulation of substrate use during the marathon.

    Science.gov (United States)

    Spriet, Lawrence L

    2007-01-01

    The energy required to run a marathon is mainly provided through oxidative phosphorylation in the mitochondria of the active muscles. Small amounts of energy from substrate phosphorylation are also required during transitions and short periods when running speed is increased. The three inputs for adenosine triphosphate production in the mitochondria include oxygen, free adenosine diphosphate and inorganic phosphate, and reducing equivalents. The reducing equivalents are derived from the metabolism of fat and carbohydrate (CHO), which are mobilised from intramuscular stores and also delivered from adipose tissue and liver, respectively. The metabolism of fat and CHO is tightly controlled at several regulatory sites during marathon running. Slower, recreational runners run at 60-65% maximal oxygen uptake (VO(2max)) for approximately 3:45:00 and faster athletes run at 70-75% for approximately 2:45:00. Both groups rely heavily on fat and CHO fuels. However, elite athletes run marathons at speeds requiring between 80% and 90% VO(2max), and finish in times between 2:05:00 and 2:20:00. They are highly adapted to oxidise fat and must do so during training. However, they compete at such high running speeds, that CHO oxidation (also highly adapted) may be the exclusive source of energy while racing. Further work with elite athletes is needed to examine this possibility.

  9. Polarimetric studies of polyethylene terephtalate flexible substrates

    Science.gov (United States)

    Stchakovsky, M.; Garcia-Caurel, E.; Warenghem, M.

    2008-12-01

    Polymer sheets are currently used worldwide in a wide range of applications. The manufacturing process of these sheets involves extruding machines that stretch the material in both lateral and longitudinal directions with respect to the machine direction, thus inducing birefringence. In most cases, the film obtained is optically biaxial. Polarimetric spectroscopy (Ellipsometry and Mueller Matrix) combined with conoscopic observation are the methods of choice to study these properties. In this work we report an analysis of commercially available polyethylene terephtalate (PET) films used as substrate for food packaging as well as for embedded electronic devices such as solar cells or flexible displays. Initial observation of these films through polarizing microscope in conoscopic mode reveals first the trace of optical axis plane with respect to the film surface and second, whether the optical axis is acute or not. This preliminary study allows optimal sample positioning for further polarimetric studies. The measurements and modelling are done in both reflection and transmission mode on several spectroscopic polarimetric setups from UV to NIR. The models give as a main result, the dielectric tensor of the film as well as its orientation with respect to the laboratory reference frame.

  10. CVD diamond substrates for electronic devices

    International Nuclear Information System (INIS)

    Holzer, H.

    1996-03-01

    In this study the applicability of chemical vapor deposition (CVD) diamond as a material for heat spreaders was investigated. Economical evaluations on the production of heat spreaders were also performed. For the diamond synthesis the hot-filament and microwave method were used respectively. The deposition parameters were varied in a way that free standing diamond layers with a thickness of 80 to 750 microns and different qualities were obtained. The influence of the deposition parameters on the relevant film properties was investigated and discussed. With both the hot-filament and microwave method it was possible to deposit diamond layers having a thermal conductivity exceeding 1200 W/mK and therefore to reach the quality level for commercial uses. The electrical resistivity was greater than 10 12 Ωcm. The investigation of the optical properties was done by Raman-, IR- and cathodoluminescence spectroscopy. Because of future applications of diamond-aluminium nitride composites as highly efficient heat spreaders diamond deposition an AIN was investigated. An improved substrate pretreatment prior to diamond deposition showed promising results for better performance of such composite heat spreaders. Both free standing layers and diamond-AIN composites could be cut by a CO2 Laser in Order to get an exact size geometry. A reduction of the diamond surface roughness was achieved by etching with manganese powder or cerium. (author)

  11. A randomised, double-blind, cross-over trial to evaluate bread, in which gluten has been pre-digested by prolyl endoprotease treatment, in subjects self-reporting benefits of adopting a gluten-free or low-gluten diet.

    Science.gov (United States)

    Rees, Dinka; Holtrop, Grietje; Chope, Gemma; Moar, Kim M; Cruickshank, Morven; Hoggard, Nigel

    2018-03-01

    The aim of the present study was to determine if the enzyme Aspergillus niger prolyl endoprotease (ANPEP), which degrades the immunogenic proline-rich residues in gluten peptides, can be used in the development of new wheat products, suitable for gluten-sensitive (GS) individuals. We have carried out a double-blind, randomised, cross-over trial with two groups of adults; subjects, self-reporting benefits of adopting a gluten-free or low-gluten diet (GS, n 16) and a control non-GS group (n 12). For the trial, volunteers consumed four wheat breads: normal bread, bread treated with 0·8 or 1 % ANPEP and low-protein bread made from biscuit flour. Compared with controls, GS subjects had a favourable cardiovascular lipid profile - lower LDL (4·0 (sem 0·3) v. 2·8 (sem 0·2) mmol/l; P=0·008) and LDL:HDL ratio (3·2 (sem 0·4) v. 1·8 (sem 0·2); P=0·005) and modified haematological profile. The majority of the GS subjects followed a low-gluten lifestyle, which helps to reduce the gastrointestinal (GI) symptoms severity. The low-gluten lifestyle does not have any effect on the quality of life, fatigue or mental state of this population. Consumption of normal wheat bread increased GI symptoms in GS subjects compared with their habitual diet. ANPEP lowered the immunogenic gluten in the treated bread by approximately 40 %. However, when compared with the control bread for inducing GI symptoms, no treatment effects were apparent. ANPEP can be applied in the production of bread with taste, texture and appearance comparable with standard bread.

  12. Hydrological performance of dual-substrate-layer green roofs using porous inert substrates with high sorption capacities.

    Science.gov (United States)

    Wang, Xiaoou; Tian, Yimei; Zhao, Xinhua; Peng, Chenrui

    2017-06-01

    Given that the common medium in existing green roofs is a single layer composed of organic and inorganic substrates, seven pilot-scale dual-substrate-layer extensive green roofs (G1-G7), which include nutrition and adsorption substrate layers, were constructed in this study. The effectiveness of porous inert substrates (activated charcoal, zeolite, pumice, lava, vermiculite and expanded perlite) used as the adsorption substrate for stormwater retention was investigated. A single-substrate-layer green roof (G8) was built for comparison with G1-G7. Despite the larger total rainfall depth (mm) of six types of simulated rains (43.2, 54.6, 76.2, 87.0, 85.2 and 86.4, respectively), the total percent retention of G1-G7 varied between 14% and 82% with an average of 43%, exhibiting better runoff-retaining capacity than G8 based on the maximum potential rainfall storage depth per unit height of adsorption substrate. Regression analysis showed that there was a logarithmic relationship between cumulative rainfall depth with non-zero runoff and stormwater retention for G1-G4 and a linear relationship for G5-G8. To enhance the water retention capacity and extend the service life of dual-substrate-layer extensive green roofs, the mixture of activated charcoal and/or pumice with expanded perlite and/or vermiculite is more suitable as the adsorption substrate than the mixture containing lava and/or zeolite.

  13. Interfacial interactions between calcined hydroxyapatite nanocrystals and substrates.

    Science.gov (United States)

    Okada, Masahiro; Furukawa, Keiko; Serizawa, Takeshi; Yanagisawa, Yoshihiko; Tanaka, Hidekazu; Kawai, Tomoji; Furuzono, Tsutomu

    2009-06-02

    Interfacial interactions between calcined hydroxyapatite (HAp) nanocrystals and surface-modified substrates were investigated by measuring adsorption behavior and adhesion strength with a quartz crystal microbalance (QCM) and a contact-mode atomic force microscope (AFM), respectively. The goal was to develop better control of HAp-nanocrystal coatings on biomedical materials. HAp nanocrystals with rodlike or spherical morphology were prepared by a wet chemical process followed by calcination at 800 degrees C with an antisintering agent to prevent the formation of sintered polycrystals. The substrate surface was modified by chemical reaction with a low-molecular-weight compound, or graft polymerization with a functional monomer. QCM measurement showed that the rodlike HAp nanocrystals adsorbed preferentially onto anionic COOH-modified substrates compared to cationic NH2- or hydrophobic CH3-modified substrates. On the other hand, the spherical nanocrystals adsorbed onto NH2- and COOH-modified substrates, which indicates that the surface properties of the HAp nanocrystals determined their adsorption behavior. The adhesion strength, which was estimated from the force required to move the nanocrystal in contact-mode AFM, on a COOH-grafted substrate prepared by graft polymerization was almost 9 times larger than that on a COOH-modified substrate prepared by chemical reaction with a low-molecular-weight compound, indicating that the long-chain polymer grafted on the substrate mitigated the surface roughness mismatch between the nanocrystal and the substrate. The adhesion strength of the nanocrystal bonded covalently by the coupling reaction to a Si(OCH3)-grafted substrate prepared by graft polymerization was approximately 1.5 times larger than that when adsorbed on the COOH-grafted substrate.

  14. An evaluation of potential dustbathing substrates for commercial broiler chickens.

    Science.gov (United States)

    Baxter, M; Bailie, C L; O'Connell, N E

    2017-12-22

    Provision of an appropriate dustbathing substrate may allow broiler chickens to satisfy a natural motivation and give them an opportunity to exercise. The main aim of this study was to evaluate the extent to which different substrates promote dustbathing behaviour in broilers. The trial was replicated over three production cycles in one commercial broiler house, with ~22 000 Ross broilers housed per cycle. The birds were provided with access to five experimental substrates from day 10 of the 6-week production cycle. The substrates included the following: (1) peat (P), (2) oat hulls (OH), (3) straw pellets (SP), (4) clean wood shavings (WS), and (5) litter control (C). The substrates were provided in 15 steel rings (1.1 m in diameter, three rings per substrate) dispersed throughout the house. The level of occupancy of the rings, behaviours performed in each substrate, and the effect of ring position (central or edge of house) were assessed in weeks 3, 4, 5 and 6 using scan sampling from video footage. Where substrates successfully promoted dustbathing, the length and components of the bouts (including number of vertical wing shakes and ground pecks) were also assessed. Results showed that birds used P significantly more than the remaining substrates for dustbathing (P0.05). The use of OH is likely to be more environmentally sustainable than that of P, and our results suggest that this substrate is relatively successful in promoting dustbathing. However, a preference was still observed for P and further work should investigate whether other suitable substrates could better reflect its qualities.

  15. Biochemical characterization of the triticale TsPAP1, a new type of plant prolyl aminopeptidase, and its impact on proline content and flowering time in transgenic Arabidopsis plants.

    Science.gov (United States)

    Zdunek-Zastocka, Edyta; Grabowska, Agnieszka; Branicki, Tomasz; Michniewska, Beata

    2017-07-01

    Proline aminopeptidase (PAP, EC 3.4.11.5) is the only enzyme that effectively releases proline from the N-termini of peptides. The amino acid sequence of the PAP from Triticosecale, TsPAP1, comprises conserved regions, characteristic of the monomeric forms of PAP found in bacteria but not yet identified in plants. Therefore, we aimed to obtain and biochemically characterize the TsPAP1 protein. The recombinant TsPAP1 protein was received through heterologous expression of the TsPAP1 coding sequence in a bacterial expression system and purified with affinity chromatography. Gel filtration chromatography and SDS electrophoresis revealed that TsPAP1 is a monomer with a molecular mass of 37.5 kDa. TsPAP1 prefers substrates with proline at the N-terminus but is also capable of hydrolyzing β-naphthylamides of hydroxyproline and alanine. Among the peptides tested, the most preferred were di- and tripeptides, especially those with glycine in the Y position. The use of diagnostic inhibitors indicated that TsPAP1 is a serine peptidase; however, further characterization revealed that the SH residues are also important for maintaining its activity. To examine the role of TsPAP1 under physiological conditions, we developed transgenic Arabidopsis plants overexpressing TsPAP1. Compared with wild-type plants, the transgenic lines accumulated more proline, flowered an average of 3.5 days earlier, and developed more siliques than did untransformed controls. Our paper is the first to describe the biochemical properties of a novel monomeric plant PAP and contributes to the functional characterization of PAP proteins in plants. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  16. Multi-substrate terpene synthases: their occurrence and physiological significance

    Directory of Open Access Journals (Sweden)

    Leila Pazouki

    2016-07-01

    Full Text Available Terpene synthases are responsible for synthesis of a large number of terpenes in plants using substrates provided by two distinct metabolic pathways, the mevalonate-dependent pathway that is located in cytosol and has been suggested to be responsible for synthesis of sesquiterpenes (C15, and 2-C-methyl-D-erythritol-4-phosphate pathway located in plastids and suggested to be responsible for the synthesis of hemi- (C5, mono- (C10 and diterpenes (C20. Recent advances in characterization of genes and enzymes responsible for substrate and end product biosynthesis as well as efforts in metabolic engineering have demonstrated existence of a number of multi-substrate terpene synthases. This review summarizes the progress in the characterization of such multi-substrate terpene synthases and suggests that the presence of multi-substrate use might have been significantly underestimated. Multi-substrate use could lead to important changes in terpene product profiles upon substrate profile changes under perturbation of metabolism in stressed plants as well as under certain developmental stages. We therefore argue that multi-substrate use can be significant under physiological conditions and can result in complicate modifications in terpene profiles.

  17. The Effect of Substrate Topography on Coating Cathodic Delamination

    DEFF Research Database (Denmark)

    Erik Weinell, Claus; Sørensen, Per A.; Kiil, Søren

    2011-01-01

    This article describes the effect of steel substrate topography on coating cathodic delamination. The study showed that the surface preparation can be used to control and minimize the rate of cathodic delamination. The coating should have maximum wetting properties so that substrates with high...

  18. A structural classification of substrate-binding proteins

    NARCIS (Netherlands)

    Berntsson, Ronnie P. -A.; Smits, Sander H. J.; Schmitt, Lutz; Slotboom, Dirk-Jan; Poolman, Bert

    2010-01-01

    Substrate-binding proteins (SBP) are associated with a wide variety of protein complexes. The proteins are part of ATP-binding cassette transporters for substrate uptake, ion gradient driven transporters, DNA-binding proteins, as well as channels and receptors from both pro-and eukaryotes. A wealth

  19. Constructed wetland using corncob charcoal substrate: pollutants removal and intensification.

    Science.gov (United States)

    Liu, Mao; Li, Boyuan; Xue, Yingwen; Wang, Hongyu; Yang, Kai

    2017-09-01

    To investigate the feasibility of using corncob charcoal substrate in constructed wetlands, four laboratory-scale vertical flow constructed wetlands (VFCWs) were built. Effluent pollutant (chemical oxygen demand (COD), NH 4 + -N, total phosphorus (TP)) concentrations during the experiment were determined to reveal pollutant removal mechanisms and efficiencies at different stages. In the stable stage, a VFCW using clay ceramisite substrate under aeration attained higher COD (95.1%), and NH 4 + -N (95.1%) removal efficiencies than a VFCW using corncob charcoal substrate (91.5% COD, 91.3% NH 4 + -N) under aeration, but lower TP removal efficiency (clay ceramisite 32.0% and corncob charcoal 40.0%). The VFCW with raw corncob substrate showed stronger COD emissions (maximum concentration 3,108 mg/L) than the corncob charcoal substrate (COD was lower than influent). The VFCW using corncob charcoal substrate performed much better than the VFCW using clay ceramisite substrate under aeration when the C/N ratio was low (C/N = 1.5, TN removal efficiency 36.89%, 4.1% respectively). These results suggest that corncob charcoal is a potential substrate in VFCWs under aeration with a unique self -supplying carbon source property in the denitrification process.

  20. Substrate inhibition kinetics of phenol degradation by binary mixed ...

    African Journals Online (AJOL)

    Steady states of a continuous culture with an inhibitory substrate were used to estimate kinetic parameters under substrate limitation (chemo stat operation). Mixed cultures of an indigenous Pseudomonas fluorescence and Pseudomonas aeruginosa were grown in continuous culture on phenol as the sole source of carbon ...

  1. Sample sufficiency of chinese pink grown in different substrates

    Directory of Open Access Journals (Sweden)

    Sidinei José lopes

    2016-04-01

    Full Text Available The cravina is an excellent plant to build up gardens due to its early flowering, abundant flowering and great performance in spring and autumn. The objective was to estimate the sample size for plant chinese pink, grown on different substrates, and check the variability of the sample size between growth parameters and production and substrates. They used seven treatments (substrates: S1 = 50% soil + 50% rice husk ash; S2 = 80% soil + 20% earthworm castings; S3 = 80% rice husk ash + 20% earthworm castings; S4 = 40% soil + 40% rice husk ash + 20% earthworm castings; S5 = 100% peat; S6 = 100% commercial substrate Mecplant®; S7 = 50% peat + 50% rice husk ash, with 56 repetitions each, totaling 392 plants of garden pink, which was evaluated in 17 of growth and production parameters. The methodology used to bootstrap resampling, with replacement, for each character within each substrate with predetermined error: 5, 10, 20 and 40% of the average (D%. To a 95% confidence interval, with D = 20%, the substrate 50% soil and 50% of rice husk ash had the largest sample size 11 characters; when comparing the characters , the number of flower buds had the highest sample size on average 113 plants. Using samples of 44 plant chinese pink for commercial substrate Mecplant® meet the lower precisions or equal to 20% for all variables. There is variation in sample size in relation to the substrate used and the variable evaluated in chinese pink plants.

  2. Development of the Enzyme-Substrate Interactions Concept Inventory

    Science.gov (United States)

    Bretz, Stacey Lowery; Linenberger, Kimberly J.

    2012-01-01

    Enzyme function is central to student understanding of multiple topics within the biochemistry curriculum. In particular, students must understand how enzymes and substrates interact with one another. This manuscript describes the development of a 15-item Enzyme-Substrate Interactions Concept Inventory (ESICI) that measures student understanding…

  3. Performance of baker's yeast produced using date syrup substrate ...

    African Journals Online (AJOL)

    Baker's yeast was produced from three selected baker's yeast strains using date syrup as a substrate at low and high flow rate compared to those produced using molasses substrates. Performance of the produced baker's yeasts on Arabic bread quality was investigated. Baking tests showed a positive relationship between ...

  4. Strawberry Production in Soilless Substrate Troughs – Plant Growth

    Science.gov (United States)

    Soilless substrates made of peat moss, coconut coir, perlite, rockwool or bark are pathogen free and they have been used in strawberry production in Europe in troughs or containers. Open field strawberry production in soilless substrate is new to California growers. The objective of this study was t...

  5. Thin film with oriented cracks on a flexible substrate

    Science.gov (United States)

    Feng, Bao; McGilvray, Andrew; Shi, Bo

    2010-07-27

    A thermoelectric film is disclosed. The thermoelectric film includes a substrate that is substantially electrically non-conductive and flexible and a thermoelectric material that is deposited on at least one surface of the substrate. The thermoelectric film also includes multiple cracks oriented in a predetermined direction.

  6. Patterned Electroplating of Micrometer Scale Magnetic Structures on Glass Substrates

    NARCIS (Netherlands)

    de Vries, A.H.B.; Kanger, Johannes S.; Krenn, Bea E.; van Driel, Roel

    2004-01-01

    This paper has developed a new method of micro patterned electroplating that enables the fabrication of micrometer scale magnetic structures on glass substrates. In contrast to other methods, the process as developed here leaves the surrounding substrate area untouched: that is there is no seed

  7. Patterned electroplating of micrometer scale magnetic structures on glass substrates.

    NARCIS (Netherlands)

    de Vries, A.H.B.; Kanger, S.J.; Krenn, G.E.; van Driel, R.

    2004-01-01

    This paper has developed a new method of micro patterned electroplating that enables the fabrication of micrometer scale magnetic structures on glass substrates. In contrast to other methods, the process as developed here leaves the surrounding substrate area untouched: that is there is no seed

  8. Rooting Rose Cuttings in Whole Pine Tree Substrates

    Science.gov (United States)

    Increased demand for alternatives to pine bark (PB) and peat moss (P) has led to extensive research on wood-based substrates, such as processed whole pine trees (WPT), for nursery and greenhouse crop production. Limited information is available on how WPT may perform as a rooting substrate for cutti...

  9. Dielectric constant of graphene-on-polarized substrate: A tight ...

    Indian Academy of Sciences (India)

    2017-06-24

    Jun 24, 2017 ... We report here a microscopic tight-binding theoretical study of the dynamic dielectric response of graphene-on-polarizable substrate with impurity. The Hamiltonian consists of first, second and third nearest neighbour electron hopping interactions besides doping and substrate-induced effects on graphene.

  10. Bioconversion of rape straw into a nutritionally enriched substrate by ...

    African Journals Online (AJOL)

    Jane

    2011-06-22

    Jun 22, 2011 ... rape straw substrate and the secretion of ligninolytic enzyme system including laccase (Lac), manganese ... results are mostly fields burning or natural degradation. The former ... Microbial conversion, especially fungal bio- conversion ... out at 27°C in plastic bags containing 200 g of lignocellulosic substrate ...

  11. Microwave flexible transistors on cellulose nanofibrillated fiber substrates

    Science.gov (United States)

    Jung-Hun Seo; Tzu-Hsuan Chang; Jaeseong Lee; Ronald Sabo; Weidong Zhou; Zhiyong Cai; Shaoqin Gong; Zhenqiang Ma

    2015-01-01

    In this paper, we demonstrate microwave flexible thin-film transistors (TFTs) on biodegradable substrates towards potential green portable devices. The combination of cellulose nanofibrillated fiber (CNF) substrate, which is a biobased and biodegradable platform, with transferrable single crystalline Si nanomembrane (Si NM), enables the realization of truly...

  12. Nanostructured surface enhanced Raman scattering substrates for explosives detection

    DEFF Research Database (Denmark)

    Schmidt, Michael Stenbaek; Olsen, Jesper Kenneth; Boisen, Anja

    2010-01-01

    Here we present a method for trace detection of explosives in the gas phase using novel surface enhanced Raman scattering (SERS) spectroscopy substrates. Novel substrates that produce an exceptionally large enhancement of the Raman effect were used to amplify the Raman signal of explosives...

  13. Substrate utilization during the first weeks of life

    NARCIS (Netherlands)

    Sauer, P. J.; Carnielli, V. P.; Sulkers, E. J.; van Goudoever, J. B.

    1994-01-01

    It is assumed that substrate utilization changes markedly around birth, from mainly glucose utilization before, to glucose/fat utilization after birth. We studied substrate oxidation and turnover in preterm infants on the first day and during the first weeks of life. We found that only part of the

  14. Effects of bamboo substrate and supplementary feed on growth and ...

    African Journals Online (AJOL)

    application as control (C), control and substrate installation (C + S) and, control and substrate with supplementary feeding (C + S + F) were randomly allotted to six earthen ponds each with an area of 100m2. Catfish fingerlings of mean weight 27.5g + 1.25 were stocked at the rate of 80 fish per 100m2. Water temperature, pH ...

  15. Design of a cylindrical LED substrate without radiator

    Science.gov (United States)

    Tang, Fan; Guo, Zhenning

    2017-12-01

    To reduce the weight and production costs of light-emitting diode (LED) lamps, we applied the principle of the chimney effect to design a cylindrical LED substrate without a radiator. We built a 3D model by using Solidworks software and applied the flow simulation plug-in to conduct model simulation, thereby optimizing the heat source distribution and substrate thickness. The results indicate that the design achieved optimal cooling with a substrate with an upper extension length of 35 mm, a lower extension length of 8 mm, and a thickness of 1 mm. For a substrate of those dimensions, the highest LED chip temperature was 64.78 °C, the weight of the substrate was 35.09 g, and R jb = 7.00 K/W. If the substrate is powered at 8, 10, and 12 W, its temperature meets LED safety requirements. In physical tests, the highest temperature for a physical 8 W cylindrical LED substrate was 66 °C, which differed by only 1.22 °C from the simulation results, verifying the validity of the simulation. The designed cylindrical LED substrate can be used in high-power LED lamps that do not require radiators. This design is not only excellent for heat dissipation, but also for its low weight, low cost, and simplicity of manufacture.

  16. Shared neural substrates of apraxia and aphasia.

    Science.gov (United States)

    Goldenberg, Georg; Randerath, Jennifer

    2015-08-01

    Apraxia is regularly associated with aphasia, but there is controversy whether their co-occurrence is the expression of a common basic deficit or results from anatomical proximity of their neural substrates. However, neither aphasia nor apraxia is an indivisible entity. Both diagnoses embrace diverse manifestations that may occur more or less independently from each other. Thus, the question whether apraxia is always accompanied by aphasia may lead to conflicting answers depending on which of their manifestations are considered. We used voxel based lesion symptom mapping (VLSM) for exploring communalities between lesion sites associated with aphasia and with apraxia. Linguistic impairment was assessed by the Aachen Aphasia Test (AAT) subtests naming, comprehension, repetition, written language, and Token Test. Apraxia was examined for imitation of meaningless hand and finger postures and for pantomime of tool use. There were two areas of overlap between aphasia and apraxia. Lesions in the anterior temporal lobe interfered with pantomime of tool use and with all linguistic tests. In the left inferior parietal lobe there was a large area where lesions were associated with defective imitation of hand postures and with poor scores on written language and the Token Test. Within this large area there were also two spots in supramarginal and angular gyrus where lesions were also associated with defective pantomime. We speculate that the coincidence of language impairment and defective pantomime after anterior temporal lesions is due to impaired access to semantic memory. The combination of defective imitation of hand postures with poor scores on Token Test and written language is not easily compatible with a crucial role of parietal regions for the conversion of concepts of intended actions into motor commands. It accords better with a role of left inferior parietal lobe regions for the categorical perception of spatial relationships. Copyright © 2015 Elsevier Ltd. All

  17. Warpage behavior analysis in package processes of embedded copper substrates

    Directory of Open Access Journals (Sweden)

    Hwang Yeong-Maw

    2017-01-01

    Full Text Available With the advance of the semiconductor industry and in response to the demands of ultra-thin products, packaging technology has been continuously developed. Thermal bonding process of copper pillar flip chip packages is a new bonding process in packaging technology, especially for substrates with embedded copper trace. During the packaging process, the substrate usually warps because of the heating process. In this paper, a finite element software ANSYS is used to model the embedded copper trace substrate and simulate the thermal and deformation behaviors of the substrate during the heating package process. A fixed geometric configuration equivalent to the real structure is duplicated to make the simulation of the warpage behavior of the substrate feasible. An empirical formula for predicting the warpage displacements is also established.

  18. Wrinkling of graphene membranes supported by silica nanoparticles on substrates

    Science.gov (United States)

    Yamamoto, Mahito; Cullen, William; Fuhrer, Michael; Einstein, Theodore; Department of Physics, University of Maryland Team

    2011-03-01

    The challenging endeavor of modulating the morphology of graphene via a patterned substrate to produce a controlled deformation has great potential importance for strain engineering the electronic properties of graphene. An essential step in this direction is to understand the response of graphene to substrate features of known geometry. Here we employ silica nanoparticles with a diameter of 10-100 nm to uniformly decorate Si O2 and mica substrates before depositing graphene, to promote nanoscale modulation of graphene geometry. The morphology of graphene on this modified substrate is then characterized by atomic force spectroscopy. We find that graphene on the substrate is locally raised by the supporting nanoparticles, and wrinkling propagates radially from the protrusions to form a ridge network which links the protrusions. We discuss the dependence of the wrinkled morphology on nanoparticle diameter and graphene thickness in terms of graphene elasticity and adhesion energy. Supported by NSF-MRSEC, Grant DMR 05-20471

  19. Substrate effects on terahertz metamaterial resonances for various metal thicknesses

    International Nuclear Information System (INIS)

    Park, S. J.; Ahn, Y. H.

    2014-01-01

    We demonstrate dielectric substrate effects on the resonance shift of terahertz metamaterials with various metal thicknesses by using finite-difference time-domain simulations. We found a small red shift in the metamaterial resonance with increasing metal thickness for the free-standing case. Conversely, when the metamaterial pattern was supported by a substrate with a high dielectric constant, the resonant frequency exhibited a large blue shift because the relative contribution of the substrate's refractive index to the resonant frequency decreased drastically as we increased the metal thickness. We determined the substrate's refractive index, 1.26, at which the metamaterial resonance was independent of the metal thickness. We extracted the effective refractive index as a function of the substrate's refractive index explicitly, which was noticeably different for different film thicknesses.

  20. Agronomic behavior of gladiolus in organic substrates with wastewater reuse

    Directory of Open Access Journals (Sweden)

    Ítalo Dourado Teixeira

    2015-12-01

    Full Text Available This study aimed to assess the technical feasibility of effluent reuse and recycling of organic substrates in the production of gladiolus. We adopted a completely randomized design in a split-plot, and the plots three qualities water (river water with Hoagland’s solution; treated effluent in stabilization pond; treated effluent in stabilization pond and disinfected and the subplots organic substrates (pine bark, coir, bagasse, repeated five times. Were characterized physico-chemical water and each substrate, the beginning and end of two growing seasons. The variables evaluated were: number of tillers, plant height, total production; flower stem length, number of flowers, time to first harvest, total cycle time, fresh and dry mass of the area. The reuse of effluent resulted in agronomic performance equal or superior to that obtained by the use of nutrient solution and the substrate base bagasse promoted agronomic performance inferior to the other substrates evaluated.

  1. Barrier layer arrangement for conductive layers on silicon substrates

    International Nuclear Information System (INIS)

    Hung, L.S.; Agostinelli, J.A.

    1990-01-01

    This patent describes a circuit element comprised of a silicon substrate and a conductive layer located on the substrate. It is characterized in that the conductive layer consists essentially of a rare earth alkaline earth copper oxide and a barrier layer triad is interposed between the silicon substrate and the conductive layer comprised of a first triad layer located adjacent the silicon substrate consisting essentially of silica, a third triad layer remote from the silicon substrate consisting essentially of a least one Group 4 heavy metal oxide, and a second triad layer interposed between the first and third triad layers consisting essentially of a mixture of silica and at lease one Group 4 heavy metal oxide

  2. The effect of grooves in amorphous substrates on the orientation of metal deposits. I - Carbon substrates

    Science.gov (United States)

    Anton, R.; Poppa, H.; Flanders, D. C.

    1982-01-01

    The graphoepitaxial alignment of vapor-deposited discrete metal crystallites is investigated in the nucleation and growth stages and during annealing by in situ UHV/TEM techniques. Various stages of nucleation, growth and coalescence of vapor deposits of Au, Ag, Pb, Sn, and Bi on amorphous, topographically structured C substrates are analyzed by advanced dark-field techniques to detect preferred local orientations. It is found that the topography-induced orientation of metal crystallites depends strongly on their mobility and their respective tendency to develop pronounced crystallographic shapes. Lowering of the average surface free energies and increasing the crystallographic surface energy anisotropies cause generally improved graphoepitaxial alignments.

  3. Reassessing SERS enhancement factors: using thermodynamics to drive substrate design.

    Science.gov (United States)

    Guicheteau, J A; Tripathi, A; Emmons, E D; Christesen, S D; Fountain, Augustus W

    2017-12-04

    Over the past 40 years fundamental and application research into Surface-Enhanced Raman Scattering (SERS) has been explored by academia, industry, and government laboratories. To date however, SERS has achieved little commercial success as an analytical technique. Researchers are tackling a variety of paths to help break through the commercial barrier by addressing the reproducibility in both the SERS substrates and SERS signals as well as continuing to explore the underlying mechanisms. To this end, investigators use a variety of methodologies, typically studying strongly binding analytes such as aromatic thiols and azarenes, and report SERS enhancement factor calculations. However a drawback of the traditional SERS enhancement factor calculation is that it does not yield enough information to understand substrate reproducibility, application potential with another analyte, or the driving factors behind the molecule-metal interaction. Our work at the US Army Edgewood Chemical Biological Center has focused on these questions and we have shown that thermodynamic principles play a key role in the SERS response and are an essential factor in future designs of substrates and applications. This work will discuss the advantages and disadvantages of various experimental techniques used to report SERS enhancement with planar SERS substrates and present our alternative SERS enhancement value. We will report on three types of analysis scenarios that all yield different information concerning the effectiveness of the SERS substrate, practical application of the substrate, and finally the thermodynamic properties of the substrate. We believe that through this work a greater understanding for designing substrates will be achieved, one that is based on both thermodynamic and plasmonic properties as opposed to just plasmonic properties. This new understanding and potential change in substrate design will enable more applications for SERS based methodologies including targeting

  4. The contribution of body composition, substrates and hormones to the variability in energy expenditure and substrate utilization in premenopausal women

    DEFF Research Database (Denmark)

    Astrup, A.; Buemann, Benjamin; Christensen, N.J.

    1992-01-01

    Twenty-four-hour energy expenditure and substrate use were measured by indirect calorimetry in respiration chambers on a fixed physical program and related to body composition and plasma concentrations of various substrates and thermogenic hormones. Fifty premenopausal women with a wide range of ...

  5. Substrate Handbook for Biogas Production; Substrathandbok foer biogasproduktion

    Energy Technology Data Exchange (ETDEWEB)

    Carlsson, My; Uldal, Martina (AnoxKaldnes AB, Lund (Sweden))

    2009-02-15

    Today, co-digestion plants in Sweden treat a broad range of different substrates, of which some have not previously been used for anaerobic digestion. The major part of this organic waste derives from households, restaurants, food industries and farms. When evaluating a new substrate as feed for anaerobic digestion, several different aspects need to be taken into consideration, such as anaerobic degradability, TS/VS content, nutrient composition and risk for mechanical problems. Consequently, there is a need for practical guidelines on how to evaluate new substrates as raw materials for biogas production, including not only gas yield but also what practical and microbiological problems that may arise when the specific substrate is treated together with other substrates in the plant. The aim with this handbook is to provide a basis on how to evaluate new substrates as feed for anaerobic digestion. The intention is that this material will save time and effort for the personnel at the plant when they come in contact with new types of waste. Also, the aim is to facilitate the process of identifying new substrates within the ABP-regulation (1774/2002) and what requirements are then demanded on handling. The work with the handbook has been divided in three different parts; (1) an extensive literature study and a compilation of the achieved results, (2) interviews with personnel at most of the Swedish co-digestion plants to identify substrates and problems of interest, and (3) lab tests of selected substrates. The lab tests included Bio Methane Potential (BMP) tests as well as a simple characterization of each substrate based on fat/protein/carbohydrate content. All data origins from anaerobic digestion within the mesophilic temperature range, but the results and discussion are applicable also for thermophilic anaerobic digestion. The result of this work is a written report together with an Excel file which are to be directly used by the biogas plants as a basis in the

  6. Comparison of filters: Inkjet printed on PEN substrate versus a laser-etched on LCP substrate

    KAUST Repository

    Arabi, Eyad A.

    2014-10-01

    In this paper, microstrip-based bandpass filters on polyethylene naphthalate (PEN) and liquid crystal polymers (LCP) are presented to investigate the performance of filters on ultra-thin substrates. PEN (with a thickness of 120 μm) has been characterized and used for a filter for the first time. In addition to being low cost and transparent, it demonstrates comparable RF performance to LCP. The conductor losses are compared by fabricating filters with inkjet printed lines as well as laser etched copper clad LCP sheets. With 5 layers of inkjet printing, and a curing temperature below 200°C, a final silver thickness of 2 μm and conductivity of 9.6 × 106 S/m are achieved. The designs are investigated at two frequencies, 24 GHz as well as 5 GHz to assess their performance at high and low frequencies respectively. The 24 GHz inkjet printed filter shows an insertion loss of 2 dB, while the 5 GHz design gives an insertion loss of 8 dB. We find that thin substrates have a strong effect on the insertion loss of filters especially as the frequency is reduced. The same design, realized on LCP (thickness of 100 μm) through laser etching, demonstrates a very similar performance, thus verifying this finding. © 2014 European Microwave Association.

  7. Comparison of filters: Inkjet printed on PEN substrate versus a laser-etched on LCP substrate

    KAUST Repository

    Arabi, Eyad A.; McKerricher, Garret; Shamim, Atif

    2014-01-01

    In this paper, microstrip-based bandpass filters on polyethylene naphthalate (PEN) and liquid crystal polymers (LCP) are presented to investigate the performance of filters on ultra-thin substrates. PEN (with a thickness of 120 μm) has been characterized and used for a filter for the first time. In addition to being low cost and transparent, it demonstrates comparable RF performance to LCP. The conductor losses are compared by fabricating filters with inkjet printed lines as well as laser etched copper clad LCP sheets. With 5 layers of inkjet printing, and a curing temperature below 200°C, a final silver thickness of 2 μm and conductivity of 9.6 × 106 S/m are achieved. The designs are investigated at two frequencies, 24 GHz as well as 5 GHz to assess their performance at high and low frequencies respectively. The 24 GHz inkjet printed filter shows an insertion loss of 2 dB, while the 5 GHz design gives an insertion loss of 8 dB. We find that thin substrates have a strong effect on the insertion loss of filters especially as the frequency is reduced. The same design, realized on LCP (thickness of 100 μm) through laser etching, demonstrates a very similar performance, thus verifying this finding. © 2014 European Microwave Association.

  8. Substrate independent approach for synthesis of graphene platelet networks

    Science.gov (United States)

    Shashurin, A.; Fang, X.; Zemlyanov, D.; Keidar, M.

    2017-06-01

    Graphene platelet networks (GPNs) comprised of randomly oriented graphene flakes two to three atomic layers thick are synthesized using a novel plasma-based approach. The approach uses a substrate capable of withstanding synthesis temperatures around 800 °C, but is fully independent of the substrate material. The synthesis occurs directly on the substrate surface without the necessity of any additional steps. GPNs were synthesized on various substrate materials including silicon (Si), thermally oxidized Si (SiO2), molybdenum (Mo), nickel (Ni) and copper (Cu), nickel-chromium (NiCr) alloy and alumina ceramics (Al2O3). The mismatch between the atomic structures of sp2 honeycomb carbon networks and the substrate material is fully eliminated shortly after the synthesis initiation, namely when about 100 nm thick deposits are formed on the substrate. GPN structures synthesized on a substrate at a temperature of about 800 °C are significantly more porous in comparison to the much denser packed amorphous carbon deposits synthesized at lower temperatures. The method proposed here can potentially revolutionize the area of electrochemical energy storage by offering a single-step direct approach for the manufacture of graphene-based electrodes for non-Faradaic supercapacitors. Mass production can be achieved using this method if a roll-to-roll system is utilized.

  9. Substrates of Peltigera Lichens as a Potential Source of Cyanobionts.

    Science.gov (United States)

    Zúñiga, Catalina; Leiva, Diego; Carú, Margarita; Orlando, Julieta

    2017-10-01

    Photobiont availability is one of the main factors determining the success of the lichenization process. Although multiple sources of photobionts have been proposed, there is no substantial evidence confirming that the substrates on which lichens grow are one of them. In this work, we obtained cyanobacterial 16S ribosomal RNA gene sequences from the substrates underlying 186 terricolous Peltigera cyanolichens from localities in Southern Chile and maritime Antarctica and compared them with the sequences of the cyanobionts of these lichens, in order to determine if cyanobacteria potentially available for lichenization were present in the substrates. A phylogenetic analysis of the sequences showed that Nostoc phylotypes dominated the cyanobacterial communities of the substrates in all sites. Among them, an overlap was observed between the phylotypes of the lichen cyanobionts and those of the cyanobacteria present in their substrates, suggesting that they could be a possible source of lichen photobionts. Also, in most cases, higher Nostoc diversity was observed in the lichens than in the substrates from each site. A better understanding of cyanobacterial diversity in lichen substrates and their relatives in the lichens would bring insights into mycobiont selection and the distribution patterns of lichens, providing a background for hypothesis testing and theory development for future studies of the lichenization process.

  10. Protein kinase substrate identification on functional protein arrays

    Directory of Open Access Journals (Sweden)

    Zhou Fang

    2008-02-01

    Full Text Available Abstract Background Over the last decade, kinases have emerged as attractive therapeutic targets for a number of different diseases, and numerous high throughput screening efforts in the pharmaceutical community are directed towards discovery of compounds that regulate kinase function. The emerging utility of systems biology approaches has necessitated the development of multiplex tools suitable for proteomic-scale experiments to replace lower throughput technologies such as mass spectroscopy for the study of protein phosphorylation. Recently, a new approach for identifying substrates of protein kinases has applied the miniaturized format of functional protein arrays to characterize phosphorylation for thousands of candidate protein substrates in a single experiment. This method involves the addition of protein kinases in solution to arrays of immobilized proteins to identify substrates using highly sensitive radioactive detection and hit identification algorithms. Results To date, the factors required for optimal performance of protein array-based kinase substrate identification have not been described. In the current study, we have carried out a detailed characterization of the protein array-based method for kinase substrate identification, including an examination of the effects of time, buffer compositions, and protein concentration on the results. The protein array approach was compared to standard solution-based assays for assessing substrate phosphorylation, and a correlation of greater than 80% was observed. The results presented here demonstrate how novel substrates for protein kinases can be quickly identified from arrays containing thousands of human proteins to provide new clues to protein kinase function. In addition, a pooling-deconvolution strategy was developed and applied that enhances characterization of specific kinase-substrate relationships and decreases reagent consumption. Conclusion Functional protein microarrays are an

  11. Microgravimetric Studies of Selenium Electrodeposition Onto Different Substrates

    Directory of Open Access Journals (Sweden)

    Kowalik R.

    2014-10-01

    Full Text Available The mechanism of selenium electrodeposition from sulfuric acid solution on different substrates was studied with the electrochemical techniques. The cyclic voltammetry was combined with the quartz crystal microbalance technique to analyze selenium deposition process. The electrochemical reduction of selenous acid on gold, silver and copper electrodes was investigated. It was found that reduction of selenous acid is a very complex process and it strongly depends from the applied substrate. The voltammetric measurements indicate the range of potentials in which the process of reduction of selenous acids on the applied substrate is possible. Additionally, the microgravimetric data confirm the deposition of selenium and they reveal the mechanism of the deposition process.

  12. Kinetic study of an alcoholic fermentation, using honey like substrate

    International Nuclear Information System (INIS)

    Gomez, Jose Angel; Castano, Hader Ivan; Arias Mario

    1997-01-01

    The paper describes a kinetic study of an alcoholic fermentation using honey like substrate; for this effect they were carried out nine fermentations in discontinuous process with a volume, of 10 L, following the behavior of the substrate concentrations, biomass and product in the time. It was evaluated the convenience of factors like the agitation and the addition of nutritious, also, it was observed the effect of the initial concentrations of substrate and inoculate and the type of honey looking for the best conditions of the process for the obtaining of an alcoholic drink

  13. Reel-to-reel substrate tape polishing system

    Energy Technology Data Exchange (ETDEWEB)

    Selvamanickam, Venkat; Gardner, Michael T.; Judd, Raymond D.; Weloth, Martin; Qiao, Yunfei

    2005-06-21

    Disclosed is a reel-to-reel single-pass mechanical polishing system (100) suitable for polishing long lengths of metal substrate tape (124) used in the manufacture of high-temperature superconductor (HTS) coated tape, including multiple instantiations of a polishing station (114) in combination with a subsequent rinsing station (116) arranged along the axis of the metal substrate tape (124) that is translating between a payout spool (110a) and a take-up spool (110b). The metal substrate tape obtains a surface smoothness that is suitable for the subsequent deposition of a buffer layer.

  14. Cell In Situ Zymography: Imaging Enzyme-Substrate Interactions.

    Science.gov (United States)

    Chhabra, Aastha; Rani, Vibha

    2017-01-01

    Zymography has long been used for the detection of substrate-specific enzyme activity. In situ zymography (ISZ), an adaptation from the conventional substrate zymography, is a widely employed technique useful for the detection, localization, and estimation of enzyme-substrate interactions in tissues. Here, we describe a protocol to detect 'in position' matrix metalloproteinase (MMP) activity in cells utilizing H9c2 cardiomyoblasts as a model. This technique is primarily adopted from the method used for histological sections and is termed as 'Cell in situ Zymography'. It is a simple, sensitive, and quantifiable methodology to assess the functional activity of an enzyme 'on site/in position' in cell culture.

  15. Adsorbates in a Box: Titration of Substrate Electronic States

    Science.gov (United States)

    Cheng, Zhihai; Wyrick, Jonathan; Luo, Miaomiao; Sun, Dezheng; Kim, Daeho; Zhu, Yeming; Lu, Wenhao; Kim, Kwangmoo; Einstein, T. L.; Bartels, Ludwig

    2010-08-01

    Nanoscale confinement of adsorbed CO molecules in an anthraquinone network on Cu(111) with a pore size of ≈4nm arranges the CO molecules in a shell structure that coincides with the distribution of substrate confined electronic states. Molecules occupy the states approximately in the sequence of rising electron energy. Despite the sixfold symmetry of the pore boundary itself, the adsorbate distribution adopts the threefold symmetry of the network-substrate system, highlighting the importance of the substrate even for such quasi-free-electron systems.

  16. Michaelis - Menten equation for degradation of insoluble substrate

    DEFF Research Database (Denmark)

    Andersen, Morten; Kari, Jeppe; Borch, Kim

    2017-01-01

    substrate it is difficult to assess whether the requirement of the MM equation is met. In this paper we study a simple kinetic model, where removal of attack sites expose new ones which preserve the total accessible substrate, and denote this approach the substrate conserving model. The kinetic equations...... are solved in closed form, both steady states and progress curves, for any admissible values of initial conditions and rate constants. The model is shown to merge with the MM equation and the reverse MM equation when these are valid. The relation between available molar concentration of attack sites and mass...

  17. Strain Multiplexed Metasurface Holograms on a Stretchable Substrate.

    Science.gov (United States)

    Malek, Stephanie C; Ee, Ho-Seok; Agarwal, Ritesh

    2017-06-14

    We demonstrate reconfigurable phase-only computer-generated metasurface holograms with up to three image planes operating in the visible regime fabricated with gold nanorods on a stretchable polydimethylsiloxane substrate. Stretching the substrate enlarges the hologram image and changes the location of the image plane. Upon stretching, these devices can switch the displayed holographic image between multiple distinct images. This work opens up the possibilities for stretchable metasurface holograms as flat devices for dynamically reconfigurable optical communication and display. It also confirms that metasurfaces on stretchable substrates can serve as platform for a variety of reconfigurable optical devices.

  18. Investigation of (111 wafers and comparison with (100 substrates

    Directory of Open Access Journals (Sweden)

    A Bahari

    2012-06-01

    Full Text Available  In the last decade, Si(100 has been used as a suitable substrate in field effect transistors. Some issues such as leakage current and tunneling current through the ultrathin films have been increased with shrinking the electronic devices – particularly, field effect transistors – to nanoscale, which is threatening more use of Si(100. We have thus demonstrated a series of experiments to grow ultrathin films on both Si(100 and Si(111 substrates and studied their nanostructural properties to see the possibility of replacing Si(100 with Si(111. The obtained results indicate that Si(111 substrate with silicon nitride film on top is desirable.

  19. Substrate-enhanced superconductivity in Li-decorated graphene

    KAUST Repository

    Kaloni, Thaneshwor P.

    2013-11-01

    We investigate the role of the substrate for the strength of the electron-phonon coupling in Li-decorated graphene. We find that the interaction with a h-BN substrate leads to a significant enhancement from to , which corresponds to a 25% increase of the transition temperature from to . The superconducting gaps amount to 1.56 meV (suspended) and 1.98 meV (supported). These findings open up a new route to enhanced superconducting transition temperatures in graphene-based materials by substrate engineering. © 2013 EPLA.

  20. Carbon Nanotube Microarrays Grown on Nanoflake Substrates

    Science.gov (United States)

    Schmidt, Howard K.; Hauge, Robert H.; Pint, Cary; Pheasant, Sean

    2013-01-01

    This innovation consists of a new composition of matter where single-walled carbon nanotubes (SWNTs) are grown in aligned arrays from nanostructured flakes that are coated in Fe catalyst. This method of growth of aligned SWNTs, which can yield well over 400 percent SWNT mass per unit substrate mass, exceeds current yields for entangled SWNT growth. In addition, processing can be performed with minimal wet etching treatments, leaving aligned SWNTs with superior properties over those that exist in entangled mats. The alignment of the nanotubes is similar to that achieved in vertically aligned nanotubes, which are called "carpets. " Because these flakes are grown in a state where they are airborne in a reactor, these flakes, after growing SWNTs, are termed "flying carpets. " These flakes are created in a roll-to-roll evaporator system, where three subsequent evaporations are performed on a 100-ft (approx. =30-m) roll of Mylar. The first layer is composed of a water-soluble "release layer, " which can be a material such as NaCl. After depositing NaCl, the second layer involves 40 nm of supporting layer material . either Al2O3 or MgO. The thickness of the layer can be tuned to synthesize flakes that are larger or smaller than those obtained with a 40-nm deposition. Finally, the third layer consists of a thin Fe catalyst layer with a thickness of 0.5 nm. The thickness of this layer ultimately determines the diameter of SWNT growth, and a layer that is too thick will result in the growth of multiwalled carbon nanotubes instead of single-wall nanotubes. However, between a thickness of 0.5 nm to 1 nm, single-walled carbon nanotubes are known to be the primary constituent. After this three-layer deposition process, the Mylar is rolled through a bath of water, which allows catalyst-coated flakes to detach from the Mylar. The flakes are then collected and dried. The method described here for making such flakes is analogous to that which is used to make birefringent ink that is

  1. Laboratory Tests of Substrate Physical Properties May Not Represent the Retention Capacity of Green Roof Substrates In Situ

    Directory of Open Access Journals (Sweden)

    Christopher Szota

    2017-11-01

    Full Text Available Green roofs can be used to reduce the volume of polluted stormwater that is generated by cities. Modelling rainfall retention is critical, but green roof water balance models often rely on the physical properties of substrates. In these models, substrate water holding capacity (WHC determines the depth of water which can be stored before runoff is generated; whereas, the permanent wilting point (PWP limits evapotranspiration. The WHC and PWP, as well as plant available water (PAW; where PAW = WHC − PWP, as determined from laboratory tests, may not truly reflect how substrates perform on green roofs. We therefore ran a simulated rainfall experiment on green roof modules to (i compare the rainfall retention of vegetated and non-vegetated substrates with different WHC and PAW, and (ii relate retention to substrate storage capacity, as calculated from laboratory measures of WHC and PAW. We found that the PAW of a substrate is a better indicator of evapotranspiration and retention when compared with WHC. However, we also found that substrates always retained less water than their calculated storage capacity would suggest, most likely being due to their high permeability. Our results indicate that using laboratory-derived measures of WHC and PAW in green roof models may be over-estimating both evapotranspiration and rainfall retention.

  2. Optical measurement of microroughness of pigment coatings on rough substrates

    Science.gov (United States)

    Elton, N. J.

    2009-02-01

    The optical determination of rms roughness at the sub-wavelength scale by measurement of specular intensity as a function of wavelength or angle of incidence is well known. The method is normally used for specimens that are macroscopically flat. However, important industrial materials such as coated paper and paint consist of microscopically rough pigment coatings on a macroscopically rough substrate. Numerical modelling is used to assess the applicability and limitations of optical measurement of microroughness for such materials. Experimental results are presented for a range of paints on substrates of various macroroughness. Model and data are in fair agreement and show that the presence of substrate macroroughness always leads to an underestimation of microroughness. Generally, optical measurements of microroughness are only comparable for substrates of similar macroroughness and a limiting value exists above which measurements may not be meaningful.

  3. Method for adhering a coating to a substrate structure

    Science.gov (United States)

    Taxacher, Glenn Curtis; Crespo, Andres Garcia; Roberts, III, Herbert Chidsey

    2015-02-17

    A method for adhering a coating to a substrate structure comprises selecting a substrate structure having an outer surface oriented substantially parallel to a direction of radial stress, modifying the outer surface to provide a textured region having steps to adhere a coating thereto, and applying a coating to extend over at least a portion of the textured region, wherein the steps are oriented substantially perpendicular to the direction of radial stress to resist deformation of the coating relative to the substrate structure. A rotating component comprises a substrate structure having an outer surface oriented substantially parallel to a direction of radial stress. The outer surface defines a textured region having steps to adhere a coating thereto, and a coating extends over at least a portion of the textured region. The steps are oriented substantially perpendicular to the direction of radial stress to resist creep.

  4. Concentration of radiocesium in cultivated mushrooms and substrates

    International Nuclear Information System (INIS)

    Miyake, Sadaaki; Higasa, Mamoru; Urabe, Ken-ichi; Haraguchi, Masato; Omura, Toshitaka

    2008-01-01

    Concentrations of 134 Cs, 137 Cs and 40 K in mushrooms cultivated in Saitama prefecture and those substrates were measured by gamma-ray spectrometry. In all mushrooms and substrates, 134 Cs was not detected. Concentrations of 137 Cs in mushrooms were in the range of 0.012 - 2.1 Bq/kg·fresh, and those in substrates were in the range of 0.080 - 1.8 Bq/kg·dry. Concentrations of 137 Cs in mushrooms varied widely with mushroom species, and the average concentration of 137 Cs in Lentinus edodes (Shiitake) was over 30 times higher than that in Pleurotus ostreatus (Hiratake). Mushroom-to-substrate concentration ratios of 137 Cs (fresh/dry) were 0.11-0.53. They were higher than the reported transfer factors for common agricultural plants. (author)

  5. Skeletal muscle substrate metabolism during exercise: methodological considerations

    DEFF Research Database (Denmark)

    Van Hall, Gerrit; González-Alonso, J; Sacchetti, M

    1999-01-01

    and the respiratory exchange ratio. However, if the aim is to quantify limb or muscle metabolism, invasive measurements have to be carried out, such as the determination of blood flow, arterio-venous (a-v) difference measurements for O2 and relevant substrates, and biopsies of the active muscle. As many substrates...... substrates. There are several methodological concerns to be aware of when studying the metabolic response to exercise in human subjects. These concerns include: (1) the muscle mass involved in the exercise is largely unknown (bicycle or treadmill). Moreover, whether the muscle sample obtained from a limb......; (3) the use of net limb glycerol release to estimate lipolysis is probably not valid (triacylglycerol utilization by muscle), since glycerol can be metabolized in skeletal muscle; (4) the precision of blood-borne substrate concentrations during exercise measured by a-v difference is hampered since...

  6. Impact and spreading behavior of cluster atoms bombarding substrates

    Energy Technology Data Exchange (ETDEWEB)

    Fang, Te-Hua, E-mail: fang.tehua@msa.hinet.net [Institute of Mechanical and Electromechanical Engineering, National Formosa University, Yunlin 632, Taiwan (China); Kang, Shao-Hui; Liao, Jia-Hung [Institute of Mechanical and Electromechanical Engineering, National Formosa University, Yunlin 632, Taiwan (China)

    2009-12-15

    The purpose of this study is to investigate the behavior of copper cluster atoms bombarding a substrate using molecule dynamics based on tight-binding second moment approximation (TB-SMA) potential. The simulated results show that a crater on the substrate surface was created by the impact of the clusters. The variations of kinetic energy of cluster bombardments can be divided into three stages. At the initial impact level, the kinetic energies of the clusters and the substrate were constant. Then, the system went into a sluggish stage of energy variation, in which the kinetic energy of the clusters reduced. In the final stage, the kinetic energy of the system became stable. The high slip vector region around the crater had a disorder damage zone. The symmetry-like cross-slip occurred beneath the top layer of the substrate along the <1 1 0> orientations. The spreading index, temperature, and potential functions that affect the bombardments are also discussed.

  7. Impact and spreading behavior of cluster atoms bombarding substrates

    International Nuclear Information System (INIS)

    Fang, Te-Hua; Kang, Shao-Hui; Liao, Jia-Hung

    2009-01-01

    The purpose of this study is to investigate the behavior of copper cluster atoms bombarding a substrate using molecule dynamics based on tight-binding second moment approximation (TB-SMA) potential. The simulated results show that a crater on the substrate surface was created by the impact of the clusters. The variations of kinetic energy of cluster bombardments can be divided into three stages. At the initial impact level, the kinetic energies of the clusters and the substrate were constant. Then, the system went into a sluggish stage of energy variation, in which the kinetic energy of the clusters reduced. In the final stage, the kinetic energy of the system became stable. The high slip vector region around the crater had a disorder damage zone. The symmetry-like cross-slip occurred beneath the top layer of the substrate along the orientations. The spreading index, temperature, and potential functions that affect the bombardments are also discussed.

  8. Highly Sensitive Filter Paper Substrate for SERS Trace Explosives Detection

    Directory of Open Access Journals (Sweden)

    Pedro M. Fierro-Mercado

    2012-01-01

    Full Text Available We report on a novel and extremely low-cost surface-enhanced Raman spectroscopy (SERS substrate fabricated depositing gold nanoparticles on common lab filter paper using thermal inkjet technology. The paper-based substrate combines all advantages of other plasmonic structures fabricated by more elaborate techniques with the dynamic flexibility given by the inherent nature of the paper for an efficient sample collection, robustness, and stability. We describe the fabrication, characterization, and SERS activity of our substrate using 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, and 1,3,5-trinitrobenzene as analytes. The paper-based SERS substrates presented a high sensitivity and excellent reproducibility for analytes employed, demonstrating a direct application in forensic science and homeland security.

  9. Substrate vibrations during courtship in three Drosophila species.

    Directory of Open Access Journals (Sweden)

    Valerio Mazzoni

    Full Text Available While a plethora of studies have focused on the role of visual, chemical and near-field airborne signals in courtship of Drosophila fruit flies, the existence of substrate-borne vibrational signals has been almost completely overlooked. Here we describe substrate vibrations generated during courtship in three species of the D. melanogaster group, from the allegedly mute species D. suzukii, its sister species D. biarmipes, and from D. melanogaster. In all species, we recorded several types of substrate vibrations which were generated by locomotion, abdominal vibrations and most likely through the activity of thoracic wing muscles. In D. melanogaster and D. suzukii, all substrate vibrations described in intact males were also recorded in males with amputated wings. Evidence suggests that vibrational signalling may be widespread among Drosophila species, and fruit flies may provide an ideal model to study various aspects of this widespread form of animal communication.

  10. Effects of Ganoderma lucidum Spent Mushroom Substrate Extract on ...

    African Journals Online (AJOL)

    Effects of Ganoderma lucidum Spent Mushroom Substrate Extract on Milk and ... Serum antioxidant activity as well as milk and serum immunoglobulin levels were ... total antioxidant capacity (TAC) and mean serum TAC concentration among ...

  11. Broadband antireflection nanodome structures on SiC substrate

    DEFF Research Database (Denmark)

    Ou, Yiyu; Zhu, Xiaolong; Møller, Uffe Visbech

    2013-01-01

    Nanodome structures are demonstrated on the SiC substrate by using nanosphere lithography and dry etching. Significant surface antireflection has been observed over a broad spectral range from 400 nm to 1600 nm....

  12. In situ synthesis of nanoparticles on substrates by inkjet printing

    KAUST Repository

    Abulikemu, Mutalifu

    2014-12-23

    Nanoparticles may be formed on a substrate by mixing precursor solutions deposited by an inkjet printer. A first solution is deposited on a substrate from a first inkjet print cartridge. Then, a second solution is deposited on the substrate from a second inkjet print cartridge. The solutions may be printed in an array of droplets on the substrate. Nanoparticles form when droplets of the first solution overlap with droplets of the second solution. In one example, the nanoparticles may be gold nanoparticles formed from mixing a first solution of 1,2-dichlorobenzene (DCB) and oleylamine and a second solution of gold chloride trihydrite and dimethyl sulfoxide (DMSO). The nanoparticles may be incorporated into optoelectronic devices.

  13. Evaluation of substrates from renewable-resources in biosurfactants ...

    African Journals Online (AJOL)

    Evaluation of substrates from renewable-resources in biosurfactants production by Pseudomonas strains. Sidnei Cerqueira dos Santos, Luzimar Gonzaga Fernandez, Juan Carlos Rossi-Alva, Milton Ricardo de Abreu Roque ...

  14. Comment on ``Ferromagnetic film on a superconducting substrate''

    Science.gov (United States)

    Sonin, E. B.

    2002-10-01

    A superconducting substrate is not able to shrink drastically domains in a ferromagnetic film, contrary to the prediction of Bulaevskii and Chudnovsky. This is shown on the basis of the exact solution for the stripe domain structure.

  15. HgCdTe photovoltaic detectors on Si substrates

    International Nuclear Information System (INIS)

    Zanio, K.R.; Bean, R.C.

    1988-01-01

    HgCdTe photovoltaic detectors have been fabricated on Si substrates through intermediate CdTe/GaAs layers. Encapsulation of the GaAs between the CdTe and Si prevents unintentional doping of the HgCdTe by Ga and As. Uniform epitaxial GaAs is grown on three inch diameter Si substrates. Detectors on such large area Si substrates will offer hybrid focal plane arrays whose dimensions are not limited by the difference between the coefficients of thermal expansion of the Si signal processor and the substrate for the HgCdTe detector array. The growth of HgCdTe detectors on the Si signal processors for monolithic focal plane arrays is also considered. 40 references

  16. In situ synthesis of nanoparticles on substrates by inkjet printing

    KAUST Repository

    Abulikemu, Mutalifu; Jabbour, Ghassan

    2014-01-01

    Nanoparticles may be formed on a substrate by mixing precursor solutions deposited by an inkjet printer. A first solution is deposited on a substrate from a first inkjet print cartridge. Then, a second solution is deposited on the substrate from a second inkjet print cartridge. The solutions may be printed in an array of droplets on the substrate. Nanoparticles form when droplets of the first solution overlap with droplets of the second solution. In one example, the nanoparticles may be gold nanoparticles formed from mixing a first solution of 1,2-dichlorobenzene (DCB) and oleylamine and a second solution of gold chloride trihydrite and dimethyl sulfoxide (DMSO). The nanoparticles may be incorporated into optoelectronic devices.

  17. Effect of substrate temperature on electrical and magnetic properties ...

    Indian Academy of Sciences (India)

    . Figure 1. The temperature dependence of resistivity for LPMO films grown at different substrate temperatures (solid and open circles are the data in zero and 1 T magnetic field). The inset shows the variation of magnetoresistance with ...

  18. Secretory Phospholipase A(2) Activity toward Diverse Substrates

    DEFF Research Database (Denmark)

    Madsen, Jesper Jonasson; Linderoth, Lars; Subramanian, Arun Kumar

    2011-01-01

    We have studied secretory phospholipase A(2)-IIA (sPLA(2)) activity toward different phospholipid analogues by performing biophysical 1 characterizations and molecular dynamics simulations. The phospholipids were natural substrates, triple alkyl phospholipids, a prodrug anticancer etherlipid, and...

  19. Screening of natural substrates and optimization of operating ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-18

    Feb 18, 2009 ... 2Department of Chemical Engineering, Eritrea Institute of Technology, Eritrea, North-East Africa. ... The work involves optimizing various parameters like substrate ... of application in food industries (Brawman, 1981; Phutela.

  20. Graphene substrates enhance optical transfection efficiency in pluripotent stem cells

    CSIR Research Space (South Africa)

    Khanyile, T

    2013-09-01

    Full Text Available Studies directed at investigating the role of nanomaterial substrates with varying properties in tissue engineering research are essential. In this research arena, pluripotent stem cells are popular for their self renewing ability and are widely...

  1. Direct transfer of graphene films for polyurethane substrate

    Energy Technology Data Exchange (ETDEWEB)

    Vilani, C.; Romani, E.C.; Larrudé, D.G. [Departamento de Física, Pontifícia Universidade Católica do Rio de Janeiro, 22451-900 Rio de Janeiro, RJ (Brazil); Barbosa, Gelza M. [Diretoria de Sistemas de Armas da Marinha, Marinha do Brasil, 20010-00 Rio de Janeiro, RJ (Brazil); Freire, F.L., E-mail: lazaro@vdg.fis.puc-rio.br [Departamento de Física, Pontifícia Universidade Católica do Rio de Janeiro, 22451-900 Rio de Janeiro, RJ (Brazil); Centro Brasileiro de Pesquisas Físicas, 22290-180 Rio de Janeiro, RJ (Brazil)

    2015-11-30

    Highlights: • Graphene was prepared by CVD using copper foils as substrates. • Monolayer, bilayer and multilayer graphene were transferred to PU. • Samples were characterized by Raman and optical spectroscopies. • PU/monolayer graphene has transmittance around 80% in visible range. - Abstract: We have proposed the direct transfer of large-area graphene films grown by chemical vapor deposition to polymeric substrate by evaporating of solvents of polyurethane/tetrahydrofurane solution. The graphene films on polyurethane substrates were characterized by Raman spectroscopy, optical and atomic force microscopies and UV–vis spectroscopy measurements. The Raman spectra revealed that it is possible to transfer in a controlled manner monolayer, bilayer and multilayer graphene films over polyurethane substrate.

  2. Hysteretic capillary condensation of 4He on Nuclepore substrates

    International Nuclear Information System (INIS)

    Godshalk, K.M.; Smith, D.T.; Hallock, R.B.

    1987-01-01

    Measurements of the approach to capillary condensation and the hysteresis encountered in capillary condensation are reported for helium adsorbed on the polycarbonate substrate Nuclepore. (Author) (5 refs., 3 figs.)

  3. Modelling of Substrate Noise and Mitigation Schemes for UWB Systems

    DEFF Research Database (Denmark)

    Shen, Ming; Mikkelsen, Jan H.; Larsen, Torben

    2012-01-01

    tuned elements in the signal paths. However, for UWB designs this is not a viable option and other means are therefore required. Moreover, owing to the ultra-wideband nature and low power spectral density of the signal, UWB mixed-signal integrated circuits are more sensitive to substrate noise compared...... with narrow-band circuits. This chapter presents a study on the modeling and mitigation of substrate noise in mixed-signal integrated circuits (ICs), focusing on UWB system/circuit designs. Experimental impact evaluation of substrate noise on UWB circuits is presented. It shows how a wide-band circuit can......The last chapter of this first part of the book, chapter seven, is devoted to Modeling of Substrate Noise and Mitigation Schemes for Ultrawideband (UWB) systems, and is written by Ming Shen, Jan H. Mikkelsen, and Torben Larsen from Aalborg University, Denmark. In highly integrated mixed...

  4. Substrate Curvature Regulates Cell Migration -A Computational Study

    Science.gov (United States)

    He, Xiuxiu; Jiang, Yi

    Cell migration in host microenvironment is essential to cancer etiology, progression and metastasis. Cellular processes of adhesion, cytoskeletal polymerization, contraction, and matrix remodeling act in concert to regulate cell migration, while local extracellular matrix architecture modulate these processes. In this work we study how stromal microenvironment with native and cell-derived curvature at micron-meter scale regulate cell motility pattern. We developed a 3D model of single cell migration on a curved substrate. Mathematical analysis of cell morphological adaption to the cell-substrate interface shows that cell migration on convex surfaces deforms more than on concave surfaces. Both analytical and simulation results show that curved surfaces regulate the cell motile force for cell's protruding front through force balance with focal adhesion and cell contraction. We also found that cell migration on concave substrates is more persistent. These results offer a novel biomechanical explanation to substrate curvature regulation of cell migration. NIH 1U01CA143069.

  5. Direct transfer of graphene films for polyurethane substrate

    International Nuclear Information System (INIS)

    Vilani, C.; Romani, E.C.; Larrudé, D.G.; Barbosa, Gelza M.; Freire, F.L.

    2015-01-01

    Highlights: • Graphene was prepared by CVD using copper foils as substrates. • Monolayer, bilayer and multilayer graphene were transferred to PU. • Samples were characterized by Raman and optical spectroscopies. • PU/monolayer graphene has transmittance around 80% in visible range. - Abstract: We have proposed the direct transfer of large-area graphene films grown by chemical vapor deposition to polymeric substrate by evaporating of solvents of polyurethane/tetrahydrofurane solution. The graphene films on polyurethane substrates were characterized by Raman spectroscopy, optical and atomic force microscopies and UV–vis spectroscopy measurements. The Raman spectra revealed that it is possible to transfer in a controlled manner monolayer, bilayer and multilayer graphene films over polyurethane substrate.

  6. Effects of mixed substrates on growth and vitamin production by ...

    African Journals Online (AJOL)

    SERVER

    2007-10-16

    Oct 16, 2007 ... The cells were grown mixotrophically in glucose (G), ethanol ... Key words: mixed substrate culture, Euglena gracilis, cell growth, vitamin production. ..... Biological elimination of nitric oxide from fuel gas by marine micro-.

  7. Pattern of diffusion-limited aggregation on nonuniform substrate

    CERN Document Server

    Ouyang Wen Ze; Zou Xian Wu; Jin Zhun Zhi

    2003-01-01

    Pattern of diffusion-limited aggregation (DLA) on nonuniform substrate was investigated by computer simulations. The nonuniform substrates are represented by Leath percolations with the probability p. p stands for the degree of nonuniformity and takes values in the range p sub c<=p<=1, where p sub c is the threshold of percolation. The DLA cluster grows up on the Leath percolation substrate. The patterns of the DLA clusters appear asymmetrical and nonuniform, and the branches are relative few for the case p is close to p sub c. In addition, the pattern depends on the shape of substrate. As p increases from p sub c to 1, cluster changes to pure DLA gradually. Correspondingly, the fractal dimension increases from 1.46 to 1.68. Also, the random walks on Leath percolations through the range p sub c<=p<=1 were examined. Our simulations show the Honda-Toyoki-Matsushita relation is still reasonable for fractional dimensional systems.

  8. Radiolabelled substrates for studying biological effects of trace contaminants

    International Nuclear Information System (INIS)

    1975-01-01

    A programme of coordinated isotopic tracer-aided investigations of the biological side-effects of foreign chemical residues in food and agriculture, initiated in 1973, was reviewed. The current status of representative investigations from the point of view of techniques and priorities was assessed. Such investigations involved radioactive substrates for studying DNA injury and its repair; 14 C-labelled acetylcholine as substrate for measuring enzyme inhibition due to the presence of, or exposure to, anticholinesteratic contaminants; radioactive substrates as indication of side-effects in non-target organisms and of their comparative susceptibilities; radioactive substrates as indicators of persistence or biodegradability of trace contaminants of soil or water; and labelled pools for studying the biological side-effects of trace contaminants. Priorities were identified

  9. Optical measurement of microroughness of pigment coatings on rough substrates

    International Nuclear Information System (INIS)

    Elton, N J

    2009-01-01

    The optical determination of rms roughness at the sub-wavelength scale by measurement of specular intensity as a function of wavelength or angle of incidence is well known. The method is normally used for specimens that are macroscopically flat. However, important industrial materials such as coated paper and paint consist of microscopically rough pigment coatings on a macroscopically rough substrate. Numerical modelling is used to assess the applicability and limitations of optical measurement of microroughness for such materials. Experimental results are presented for a range of paints on substrates of various macroroughness. Model and data are in fair agreement and show that the presence of substrate macroroughness always leads to an underestimation of microroughness. Generally, optical measurements of microroughness are only comparable for substrates of similar macroroughness and a limiting value exists above which measurements may not be meaningful

  10. Identifying 8-hydroxynaringenin as a suicide substrate of mushroom tyrosinase.

    Science.gov (United States)

    Chang, Te-Sheng; Lin, Meng-Yi; Lin, Hsuan-Jung

    2010-01-01

    A biotransformed metabolite of naringenin was isolated from the fermentation broth of Aspergillus oryzae, fed with naringenin, and identified as 8-hydroxynaringenin based on the mass and (1)H- and (13)C-NMR spectral data. The compound showed characteristics of both an irreversible inhibitor and a substrate of mushroom tyrosinase in preincubation and HPLC analysis. These results demonstrate that 8-hydroxynaringenin belongs to a suicide substrate of mushroom tyrosinase. The partition ratio between the compound's molecules in the formation of product and in the inactivation of the enzyme was determined to be 283 +/- 21. The present study's results, together with our previous findings, which proved that both 8-hydroxydaidzein and 8-hydroxygenistein are suicide substrates of mushroom tyrosinase, show that 7,8,4'-trihydroxyl functional groups on flavonoids' skeletons play important roles in producing suicide substrate properties toward mushroom tyrosinase.

  11. Substrate heating and cooling during magnetron sputtering of copper target

    Energy Technology Data Exchange (ETDEWEB)

    Shapovalov, Viktor I.; Komlev, Andrey E.; Bondarenko, Anastasia S., E-mail: stopnastia@gmail.com; Baykov, Pavel B.; Karzin, Vitaliy V.

    2016-02-22

    Heating and cooling processes of the substrate during the DC magnetron sputtering of the copper target were investigated. The sensitive element of a thermocouple was used as a substrate. It was found, that the heat outflow rate from the substrate is lower when the magnetron is turned off rather than when it is turned on. Furthermore, the heating rate, the ultimate temperature, and the heat outflow rate related to the deposition of copper atoms are directly proportional to the discharge current density. - Highlights: • New effect of heat outflow from substrate when magnetron is on was discovered. • This new effect is linear in terms of heat outflow rate to target current ratio. • Kinetic equation for heating process additively considers this effect.

  12. Design and modeling of Faraday cages for substrate noise isolation

    Science.gov (United States)

    Wu, Joyce H.; del Alamo, Jesús A.

    2013-07-01

    A Faraday cage structure using through-substrate vias is an effective strategy to suppress substrate crosstalk, particularly at high frequencies. Faraday cages can reduce substrate noise by 32 dB at 10 GHz, and 26 dB at 50 GHz. We have developed lumped-element, equivalent circuit models of the Faraday cages and test structures to better understand the performance of the Faraday cages. These models compare well to measured results and show that the vias of the Faraday cage act as an RLC shunt to ground that draws substrate current. Designing a Faraday cage to achieve optimum isolation requires low via impedance and mitigation of via sidewall capacitance. The Faraday cage inductance is correlated to the number of vias and via spacing of the cage and can be optimized for the frequency of operation.

  13. Substrate utilization and VSS relations in activated sludge processes

    Energy Technology Data Exchange (ETDEWEB)

    Droste, R.L.; Fernandes, L.; Sun, X. [Ottawa Univ., ON (Canada). Dept. of Civil Engineering

    1993-12-31

    A new empirical substrate removal model for activated sludge in continuous flow stirred tank reactor (CFSTR) and sequencing batch reactor (SBR) was developed in this study. This model includes an exponential function of volatile suspended solids to express the active biomass which is actually involved in substrate utilization. Results indicate that the proposed exponential models predict more accurately effluent COD in CFSTR and SBR systems than the first or zero order models. (author). 7 refs., 1 fig., 4 tabs.

  14. Coating-substrate-simulations applied to HFQ® forming tools

    Directory of Open Access Journals (Sweden)

    Leopold Jürgen

    2015-01-01

    Full Text Available In this paper a comparative analysis of coating-substrate simulations applied to HFQTM forming tools is presented. When using the solution heat treatment cold die forming and quenching process, known as HFQTM, for forming of hardened aluminium alloy of automotive panel parts, coating-substrate-systems have to satisfy unique requirements. Numerical experiments, based on the Advanced Adaptive FE method, will finally present.

  15. Substrate specificity determinants of class III nucleotidyl cyclases.

    Science.gov (United States)

    Bharambe, Nikhil G; Barathy, Deivanayaga V; Syed, Wajeed; Visweswariah, Sandhya S; Colaςo, Melwin; Misquith, Sandra; Suguna, Kaza

    2016-10-01

    The two second messengers in signalling, cyclic AMP and cyclic GMP, are produced by adenylyl and guanylyl cyclases respectively. Recognition and discrimination of the substrates ATP and GTP by the nucleotidyl cyclases are vital in these reactions. Various apo-, substrate- or inhibitor-bound forms of adenylyl cyclase (AC) structures from transmembrane and soluble ACs have revealed the catalytic mechanism of ATP cyclization reaction. Previously reported structures of guanylyl cyclases represent ligand-free forms and inactive open states of the enzymes and thus do not provide information regarding the exact mode of substrate binding. The structures we present here of the cyclase homology domain of a class III AC from Mycobacterium avium (Ma1120) and its mutant in complex with ATP and GTP in the presence of calcium ion, provide the structural basis for substrate selection by the nucleotidyl cyclases at the atomic level. Precise nature of the enzyme-substrate interactions, novel modes of substrate binding and the ability of the binding pocket to accommodate diverse conformations of the substrates have been revealed by the present crystallographic analysis. This is the first report to provide structures of both the nucleotide substrates bound to a nucleotidyl cyclase. Coordinates and structure factors have been deposited in the Protein Data Bank with accession numbers: 5D15 (Ma1120 CHD +ATP.Ca 2+ ), 5D0E (Ma1120 CHD +GTP.Ca 2+ ), 5D0H (Ma1120 CHD (KDA→EGY)+ATP.Ca 2+ ), 5D0G (Ma1120 CHD (KDA→EGY)+GTP.Ca 2+ ). Adenylyl cyclase (EC number: 4.6.1.1). © 2016 Federation of European Biochemical Societies.

  16. Ultrasensitivity in phosphorylation-dephosphorylation cycles with little substrate.

    Directory of Open Access Journals (Sweden)

    Bruno M C Martins

    Full Text Available Cellular decision-making is driven by dynamic behaviours, such as the preparations for sunrise enabled by circadian rhythms and the choice of cell fates enabled by positive feedback. Such behaviours are often built upon ultrasensitive responses where a linear change in input generates a sigmoidal change in output. Phosphorylation-dephosphorylation cycles are one means to generate ultrasensitivity. Using bioinformatics, we show that in vivo levels of kinases and phosphatases frequently exceed the levels of their corresponding substrates in budding yeast. This result is in contrast to the conditions often required by zero-order ultrasensitivity, perhaps the most well known means for how such cycles become ultrasensitive. We therefore introduce a mechanism to generate ultrasensitivity when numbers of enzymes are higher than numbers of substrates. Our model combines distributive and non-distributive actions of the enzymes with two-stage binding and concerted allosteric transitions of the substrate. We use analytical and numerical methods to calculate the Hill number of the response. For a substrate with [Formula: see text] phosphosites, we find an upper bound of the Hill number of [Formula: see text], and so even systems with a single phosphosite can be ultrasensitive. Two-stage binding, where an enzyme must first bind to a binding site on the substrate before it can access the substrate's phosphosites, allows the enzymes to sequester the substrate. Such sequestration combined with competition for each phosphosite provides an intuitive explanation for the sigmoidal shifts in levels of phosphorylated substrate. Additionally, we find cases for which the response is not monotonic, but shows instead a peak at intermediate levels of input. Given its generality, we expect the mechanism described by our model to often underlay decision-making circuits in eukaryotic cells.

  17. Ultrasound-Enhanced Biogas Production from Different Substrates

    DEFF Research Database (Denmark)

    González-Fernández, Cristina; Timmers, Rudolphus Antonius; Ruiz, Begona

    2015-01-01

    Among the biofuel production processes using different substrates, the biogas generation process is one of the simplest. Compared with bioethanol or biodiesel production processes, anaerobic digestion is a process where all the organic matter (carbohydrates, lipids and proteins) can be biologically...... production. The present chapter is dedicated to providing a review of ultrasound pretreatment applied to different substrates (lignocelullosic materials, manures, sludge and microalgae). The advantages and constraints, that ultrasound pretreatment exhibit towards biogas production, are discussed and compared...

  18. Preparation of surface enhanced Raman substrate and its characterization

    Science.gov (United States)

    Liu, Y.; Wang, J. Y.; Wang, J. Q.

    2017-10-01

    Surface enhanced Raman spectroscopy (SERS) is a fast, convenient and highly sensitive detection technique, and preparing the good effect and repeatable substrate is the key to realize the trace amount and quantitative detection in the field of food safety detection. In this paper, a surface enhanced Raman substrate based on submicrometer silver particles structure was prepared by chemical deposition method, and characterized its structure and optical properties.

  19. Substrate utilization and VSS relations in activated sludge processes

    Energy Technology Data Exchange (ETDEWEB)

    Droste, R L; Fernandes, L; Sun, X [Ottawa Univ., ON (Canada). Dept. of Civil Engineering

    1994-12-31

    A new empirical substrate removal model for activated sludge in continuous flow stirred tank reactor (CFSTR) and sequencing batch reactor (SBR) was developed in this study. This model includes an exponential function of volatile suspended solids to express the active biomass which is actually involved in substrate utilization. Results indicate that the proposed exponential models predict more accurately effluent COD in CFSTR and SBR systems than the first or zero order models. (author). 7 refs., 1 fig., 4 tabs.

  20. Adhesive interactions of geckos with wet and dry fluoropolymer substrates.

    Science.gov (United States)

    Stark, Alyssa Y; Dryden, Daniel M; Olderman, Jeffrey; Peterson, Kelly A; Niewiarowski, Peter H; French, Roger H; Dhinojwala, Ali

    2015-07-06

    Fluorinated substrates like Teflon® (poly(tetrafluoroethylene); PTFE) are well known for their role in creating non-stick surfaces. We showed previously that even geckos, which can stick to most surfaces under a wide variety of conditions, slip on PTFE. Surprisingly, however, geckos can stick reasonably well to PTFE if it is wet. In an effort to explain this effect, we have turned our attention to the role of substrate surface energy and roughness when shear adhesion occurs in media other than air. In this study, we removed the roughness component inherent to commercially available PTFE and tested geckos on relatively smooth wet and dry fluoropolymer substrates. We found that roughness had very little effect on shear adhesion in air or in water and that the level of fluorination was most important for shear adhesion, particularly in air. Surface energy calculations of the two fluorinated substrates and one control substrate using the Tabor-Winterton approximation and the Young-Dupré equation were used to determine the interfacial energy of the substrates. Using these interfacial energies we estimated the ratio of wet and dry normal adhesion for geckos clinging to the three substrates. Consistent with the results for rough PTFE, our predictions show a qualitative trend in shear adhesion based on fluorination, and the quantitative experimental differences highlight the unusually low shear adhesion of geckos on dry smooth fluorinated substrates, which is not captured by surface energy calculations. Our work has implications for bioinspired design of synthetics that can preferentially stick in water but not in air.

  1. Structural basis for substrate specificities of cellular deoxyribonucleoside kinases

    DEFF Research Database (Denmark)

    Johansson, K.; Ramaswamy, S.; Ljungcrantz, C.

    2001-01-01

    Deoxyribonucleoside kinases phosphorylate deoxyribonucleosides and activate a number of medically important nucleoside analogs. Here we report the structure of the Drosophila deoxyribonucleoside kinase with deoxycytidine bound at the nucleoside binding site and that of the human deoxyguanosine ki......; this is apparently due to the presence of Arg 118, which provides favorable hydrogen bonding interactions with the substrate. The two new structures provide an explanation for the substrate specificity of cellular deoxyribonucleoside kinases....

  2. Chemically Functionalized Carbon Nanotubes as Substrates for Neuronal Growth

    Science.gov (United States)

    Hu, Hui; Ni, Yingchun; Montana, Vedrana; Haddon, Robert C.; Parpura, Vladimir

    2009-01-01

    We report the use of chemically modified carbon nanotubes as a substrate for cultured neurons. The morphological features of neurons that directly reflect their potential capability in synaptic transmission are characterized. The chemical properties of carbon nanotubes are systematically varied by attaching different functional groups that confer known characteristics to the substrate. By manipulating the charge carried by functionalized carbon nanotubes we are able to control the outgrowth and branching pattern of neuronal processes. PMID:21394241

  3. Colloidal quantum dot solar cells on curved and flexible substrates

    International Nuclear Information System (INIS)

    Kramer, Illan J.; Moreno-Bautista, Gabriel; Minor, James C.; Kopilovic, Damir; Sargent, Edward H.

    2014-01-01

    Colloidal quantum dots (CQDs) are semiconductor nanocrystals synthesized with, processed in, and deposited from the solution phase, potentially enabling low-cost, facile manufacture of solar cells. Unfortunately, CQD solar cell reports, until now, have only explored batch-processing methods—such as spin-coating—that offer limited capacity for scaling. Spray-coating could offer a means of producing uniform colloidal quantum dot films that yield high-quality devices. Here, we explore the versatility of the spray-coating method by producing CQD solar cells in a variety of previously unexplored substrate arrangements. The potential transferability of the spray-coating method to a roll-to-roll manufacturing process was tested by spray-coating the CQD active layer onto six substrates mounted on a rapidly rotating drum, yielding devices with an average power conversion efficiency of 6.7%. We further tested the manufacturability of the process by endeavoring to spray onto flexible substrates, only to find that spraying while the substrate was flexed was crucial to achieving champion performance of 7.2% without compromise to open-circuit voltage. Having deposited onto a substrate with one axis of curvature, we then built our CQD solar cells onto a spherical lens substrate having two axes of curvature resulting in a 5% efficient device. These results show that CQDs deposited using our spraying method can be integrated to large-area manufacturing processes and can be used to make solar cells on unconventional shapes.

  4. Cracking in thin films of colloidal particles on elastomeric substrates

    Science.gov (United States)

    Smith, Michael; Sharp, James

    2012-02-01

    The drying of thin colloidal films of particles is a common industrial problem (e.g paint drying, ceramic coatings). An often undesirable side effect is the appearance of cracks. As the liquid in a suspension evaporates, particles are forced into contact both with each other and the substrate, forming a fully wetted film. Under carefully controlled conditions the observed cracks grow orthogonal to the drying front, spaced at regular intervals along it. In this work we investigated the role of the substrate in constraining the film. Atomic force microscopy, was used to image the particle arrangements on the top and bottom surfaces of films, dried on liquid and glass substrates. We present convincing evidence that the interface prevents particle rearrangements at the bottom of the film, leading to a mismatch strain between upper and lower surfaces of the film which appears to drive cracking. We show that when the modulus of the substrate becomes comparable to the stresses measured in the films, the crack spacing is significantly altered. We also show that cracks do not form on liquid substrates. These combined experiments highlight the importance of substrate constraint in the crack formation mechanism.[4pt] [1] M.I. Smith, J.S. Sharp, Langmuir 27, 8009 (2011)

  5. Characterizing Protease Specificity: How Many Substrates Do We Need?

    Directory of Open Access Journals (Sweden)

    Michael Schauperl

    Full Text Available Calculation of cleavage entropies allows to quantify, map and compare protease substrate specificity by an information entropy based approach. The metric intrinsically depends on the number of experimentally determined substrates (data points. Thus a statistical analysis of its numerical stability is crucial to estimate the systematic error made by estimating specificity based on a limited number of substrates. In this contribution, we show the mathematical basis for estimating the uncertainty in cleavage entropies. Sets of cleavage entropies are calculated using experimental cleavage data and modeled extreme cases. By analyzing the underlying mathematics and applying statistical tools, a linear dependence of the metric in respect to 1/n was found. This allows us to extrapolate the values to an infinite number of samples and to estimate the errors. Analyzing the errors, a minimum number of 30 substrates was found to be necessary to characterize substrate specificity, in terms of amino acid variability, for a protease (S4-S4' with an uncertainty of 5 percent. Therefore, we encourage experimental researchers in the protease field to record specificity profiles of novel proteases aiming to identify at least 30 peptide substrates of maximum sequence diversity. We expect a full characterization of protease specificity helpful to rationalize biological functions of proteases and to assist rational drug design.

  6. Colloidal quantum dot solar cells on curved and flexible substrates

    KAUST Repository

    Kramer, Illan J.; Moreno-Bautista, Gabriel; Minor, James C.; Kopilovic, Damir; Sargent, Edward H.

    2014-01-01

    © 2014 AIP Publishing LLC. Colloidal quantum dots (CQDs) are semiconductor nanocrystals synthesized with, processed in, and deposited from the solution phase, potentially enabling low-cost, facile manufacture of solar cells. Unfortunately, CQD solar cell reports, until now, have only explored batch-processing methods - such as spin-coating - that offer limited capacity for scaling. Spray-coating could offer a means of producing uniform colloidal quantum dot films that yield high-quality devices. Here, we explore the versatility of the spray-coating method by producing CQD solar cells in a variety of previously unexplored substrate arrangements. The potential transferability of the spray-coating method to a roll-to-roll manufacturing process was tested by spray-coating the CQD active layer onto six substrates mounted on a rapidly rotating drum, yielding devices with an average power conversion efficiency of 6.7%. We further tested the manufacturability of the process by endeavoring to spray onto flexible substrates, only to find that spraying while the substrate was flexed was crucial to achieving champion performance of 7.2% without compromise to open-circuit voltage. Having deposited onto a substrate with one axis of curvature, we then built our CQD solar cells onto a spherical lens substrate having two axes of curvature resulting in a 5% efficient device. These results show that CQDs deposited using our spraying method can be integrated to large-area manufacturing processes and can be used to make solar cells on unconventional shapes.

  7. Energetic efficiency of complex substrate utilization by Trichoderma viride

    Energy Technology Data Exchange (ETDEWEB)

    Leite, M; Apine, A; Zeltina, M; Shvinka, J [AN Latvijskoj SSR, Riga (USSR). August Kirchstein Inst. of Microbiology

    1989-01-01

    The efficiency of carbon substrate utilization is evaluated as the thermodynamic efficiency (eta{sub x}) of microbial growth. Three methods based on mass-energy balance are used for the efficiency studies of complex substrates (straw, plant juices, lye) utilization by microfungi Trichoderma viride. 1. According to substrate and biomass balance eta{sub x}=0.55, 0.37 and 0.36 for Trichoderma viride growth on alkali pretreated wheat straw during 23, 34 and 50 hours. Cellulose biodegradation increases with cultivation time. However, the efficiency of cellulose utilization for cell mass growth decreases at the same time. 2. In accordance with oxygen-balance calculations eta{sub x}=0.75 and 0.71 for the same processes. The discrepancy in results from the above two methods probably can be explained by the following: A. Substrate and biomass balance gives underestimated results. B. Oxygen balance method includes the part of energy for extracellular product formation and therefore eta{sub x} can be overestimated. C. The efficiency of complex soluble substrate utilization (lye, green juice, deproteinized brown plant juice) tested by means of pulse method gives the values of eta{sub x}=0.72-0.88. Similar high estimates of eta{sub x} in C-limited batch culture are observed for soluble carbohydrates (glucose, galactose, lactose, xylose) but not for acetate. The pulse method is advantageous for testing the 'true' efficiency of carbon substrate utilization in a definite physiological environment. (orig.).

  8. Colloidal quantum dot solar cells on curved and flexible substrates

    KAUST Repository

    Kramer, Illan J.

    2014-10-20

    © 2014 AIP Publishing LLC. Colloidal quantum dots (CQDs) are semiconductor nanocrystals synthesized with, processed in, and deposited from the solution phase, potentially enabling low-cost, facile manufacture of solar cells. Unfortunately, CQD solar cell reports, until now, have only explored batch-processing methods - such as spin-coating - that offer limited capacity for scaling. Spray-coating could offer a means of producing uniform colloidal quantum dot films that yield high-quality devices. Here, we explore the versatility of the spray-coating method by producing CQD solar cells in a variety of previously unexplored substrate arrangements. The potential transferability of the spray-coating method to a roll-to-roll manufacturing process was tested by spray-coating the CQD active layer onto six substrates mounted on a rapidly rotating drum, yielding devices with an average power conversion efficiency of 6.7%. We further tested the manufacturability of the process by endeavoring to spray onto flexible substrates, only to find that spraying while the substrate was flexed was crucial to achieving champion performance of 7.2% without compromise to open-circuit voltage. Having deposited onto a substrate with one axis of curvature, we then built our CQD solar cells onto a spherical lens substrate having two axes of curvature resulting in a 5% efficient device. These results show that CQDs deposited using our spraying method can be integrated to large-area manufacturing processes and can be used to make solar cells on unconventional shapes.

  9. Flexible Bragg reflection waveguide devices fabricated on a plastic substrate

    Science.gov (United States)

    Kim, Kyung-Jo; Yi, Jeong-Ah; Oh, Min-Cheol; Noh, Young-Ouk; Lee, Hyung-Jong

    2007-09-01

    Bragg reflecting waveguide devices are fabricated on a flexible substrate by using a post lift-off process in order to provide highly uniform grating patterns on a wide range. In this process, the flexible substrate spin-coated on silicon wafer is released after the final fabrication process of chip dicing. The fabricated flexible Bragg reflector shows very sharp transmission spectrum with 3-dB bandwidth of 0.1 nm and 10-dB bandwidth of 0.4 nm, which proves the Bragg reflector has excellent uniformity. To achieve athermal operation of the flexible Bragg reflector, thermal expansion property of the plastic substrate is controlled by the thickness of two polymer materials constructing the plastic substrate. The flexible substrate with 0.7-μm SU-8 layers sandwiching 100-μm NOA61 layer provides an optimized thermal expansion property to compensate the thermo-optic effect of the waveguide made of ZPU polymer. The temperature dependence of the Bragg reflector is decreased to -0.011 nm/°C through the incorporation of the plastic substrate.

  10. Substrate channel in nitrogenase revealed by a molecular dynamics approach.

    Science.gov (United States)

    Smith, Dayle; Danyal, Karamatullah; Raugei, Simone; Seefeldt, Lance C

    2014-04-15

    Mo-dependent nitrogenase catalyzes the biological reduction of N2 to two NH3 molecules at FeMo-cofactor buried deep inside the MoFe protein. Access of substrates, such as N2, to the active site is likely restricted by the surrounding protein, requiring substrate channels that lead from the surface to the active site. Earlier studies on crystallographic structures of the MoFe protein have suggested three putative substrate channels. Here, we have utilized submicrosecond atomistic molecular dynamics simulations to allow the nitrogenase MoFe protein to explore its conformational space in an aqueous solution at physiological ionic strength, revealing a putative substrate channel. The viability of this observed channel was tested by examining the free energy of passage of N2 from the surface through the channel to FeMo-cofactor, resulting in the discovery of a very low energy barrier. These studies point to a viable substrate channel in nitrogenase that appears during thermal motions of the protein in an aqueous environment and that approaches a face of FeMo-cofactor earlier implicated in substrate binding.

  11. Acclimatization and growth of ornamental pineapple seedlings under organic substrates

    Directory of Open Access Journals (Sweden)

    Ronan Carlos Colombo

    2017-09-01

    Full Text Available The in vitro propagation techniques are commonly used to produce ornamental pineapple seedlings in commercial scale, aiming to attend the growers with genetic and sanitary quality seedlings. However, the choice of the ideal substrate is essential for the acclimatization and growth stage of the seedlings propagated by this technique, since some substrates can increase the seedling mortality and/or limit the seedling growth due to its physical and chemical characteristics. Thus, the aim of this study was to evaluate the acclimatization of ornamental pineapple [Ananas comosus (L. Merr. var. ananassoides (Baker Coppens & Leal] on different substrates. Seedlings with approximately seven centimeters, obtained from in vitro culture, were transplanted into styrofoam trays filled with the following substrates: sphagnum; semi-composed pine bark; carbonized rice husk; sphagnum + semicomposed pine bark; sphagnum + carbonized rice husk; and semi-composed pine bark + carbonized rice husk. Each treatment was replicated five times using 10 plants. At 180 days, there were evaluated the following variables: survival percentage, plant height, number of leaves, leaf area, largest root length, and shoot and root dry matter. The substrate semi-composed pine bark + carbonized rice husk presented the lowest mean (62% for survival percentage. The semi-composed pine bark and semi-composed pine bark + carbonized rice husk treatments presented significant increments in some evaluated biometric characteristics. The semi-composed pine bark is the most favorable substrate for the A. comosus var. ananassoids acclimatization.

  12. Caged Protein Prenyltransferase Substrates: Tools for Understanding Protein Prenylation

    Energy Technology Data Exchange (ETDEWEB)

    DeGraw, Amanda J.; Hast, Michael A.; Xu, Juhua; Mullen, Daniel; Beese, Lorena S.; Barany, George; Distefano, Mark D. (Duke); (UMM)

    2010-11-15

    Originally designed to block the prenylation of oncogenic Ras, inhibitors of protein farnesyltransferase currently in preclinical and clinical trials are showing efficacy in cancers with normal Ras. Blocking protein prenylation has also shown promise in the treatment of malaria, Chagas disease and progeria syndrome. A better understanding of the mechanism, targets and in vivo consequences of protein prenylation are needed to elucidate the mode of action of current PFTase (Protein Farnesyltransferase) inhibitors and to create more potent and selective compounds. Caged enzyme substrates are useful tools for understanding enzyme mechanism and biological function. Reported here is the synthesis and characterization of caged substrates of PFTase. The caged isoprenoid diphosphates are poor substrates prior to photolysis. The caged CAAX peptide is a true catalytically caged substrate of PFTase in that it is to not a substrate, yet is able to bind to the enzyme as established by inhibition studies and X-ray crystallography. Irradiation of the caged molecules with 350 nm light readily releases their cognate substrate and their photolysis products are benign. These properties highlight the utility of those analogs towards a variety of in vitro and in vivo applications.

  13. Effect of Substrate Permittivity and Thickness on Performance of Single-Layer, Wideband, U-Slot Antennas on Microwave Substrates

    National Research Council Canada - National Science Library

    Natarajan, V; Chatterjee, D

    2004-01-01

    This paper presents effects of substrate permittivity and thickness on the performance characteristics like impedance bandwidth, radiation efficiency and gain of a single-layer, wideband, U-slot antenna...

  14. Substrate deposit effect on the characteristic of an intertidal macroalgal community

    Digital Repository Service at National Institute of Oceanography (India)

    Imchen, T.

    Present study consists the effect of substrate deposit (silt, clay, sand, gravel and shards of shells) on the characteristic of an intertidal rocky shore macroalgae Macroalgal assemblage was segregated from substrate deposit in two stages Substrate...

  15. Single-molecule FRET unveils induced-fit mechanism for substrate selectivity in flap endonuclease 1

    KAUST Repository

    Rashid, Fahad; Harris, Paul D.; Zaher, Manal S.; Sobhy, Mohamed Abdelmaboud; Joudeh, Luay I; Yan, Chunli; Piwonski, Hubert Marek; Tsutakawa, Susan E; Ivanov, Ivaylo; Tainer, John A; Habuchi, Satoshi; Hamdan, Samir

    2017-01-01

    that efficiently verifies substrate and suppresses off-target cleavage. FEN1 sculpts DNA with diffusion-limited kinetics to test DNA substrate. This DNA distortion mutually 'locks' protein and DNA conformation and enables substrate verification with extreme

  16. Identification of putative substrates for cynomolgus monkey cytochrome P450 2C8 by substrate depletion assays with 22 human P450 substrates and inhibitors.

    Science.gov (United States)

    Hosaka, Shinya; Murayama, Norie; Satsukawa, Masahiro; Uehara, Shotaro; Shimizu, Makiko; Iwasaki, Kazuhide; Iwano, Shunsuke; Uno, Yasuhiro; Yamazaki, Hiroshi

    2016-07-01

    Cynomolgus monkeys are widely used in drug developmental stages as non-human primate models. Previous studies used 89 compounds to investigate species differences associated with cytochrome P450 (P450 or CYP) function that reported monkey specific CYP2C76 cleared 19 chemicals, and homologous CYP2C9 and CYP2C19 metabolized 17 and 30 human CYP2C9 and/or CYP2C19 substrates/inhibitors, respectively. In the present study, 22 compounds selected from viewpoints of global drug interaction guidances and guidelines were further evaluated to seek potential substrates for monkey CYP2C8, which is highly homologous to human CYP2C8 (92%). Amodiaquine, montelukast, quercetin and rosiglitazone, known as substrates or competitive inhibitors of human CYP2C8, were metabolically depleted by recombinant monkey CYP2C8 at relatively high rates. Taken together with our reported findings of the slow eliminations of amodiaquine and montelukast by monkey CYP2C9, CYP2C19 and CYP2C76, the present results suggest that these at least four chemicals may be good marker substrates for monkey CYP2C8. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  17. Real-time sensing of epithelial cell-cell and cell-substrate interactions by impedance spectroscopy on porous substrates

    Energy Technology Data Exchange (ETDEWEB)

    Mondal, D.; RoyChaudhuri, C., E-mail: chirosreepram@yahoo.com [Department of Electronics and Telecommunication Engineering, Indian Institute of Engineering Science and Technology, Shibpur, Howrah 711103 (India); Pal, D. [Department of Aerospace Engineering and Applied Mechanics, Indian Institute of Engineering Science and Technology, Shibpur, Howrah 711103 (India)

    2015-07-28

    Oxidized porous silicon (PS) is a common topographical biocompatible substrate that potentially provides a distinct in vitro environment for better understanding of in vivo behavior. But in the reported studies on oxidized PS, cell-cell and cell-substrate interactions have been detected only by fluorescent labeling. This paper is the first attempt to investigate real-time sensing of these interactions on HaCaT cells by label-free impedance spectroscopy on oxidized PS of two pore diameters (50 and 500 nm). One of the major requirements for successful impedance spectroscopy measurement is to restrict the channeling of electric field lines through the pores. To satisfy this criterion, we have designed the pore depths after analyzing the penetration of the medium by using computational fluid dynamics simulation. A distributed electrical model was also developed for estimating the various cellular attributes by considering a pseudorandom distribution of pores. It is observed from the impedance measurements and from the model that the proliferation rate increases for 50 nm pores but decreases for 500 nm pores compared to that for planar substrates. The rate of decrease in cell substrate separation (h) in the initial stage is more than the rate of increase in cell-cell junction resistance (R{sub b}) corresponding to the initial adhesion phase of cells. It is observed that R{sub b} and h are higher for 50 nm pores than those for planar substrates, corresponding to the fact that substrates more conducive toward cell adhesion encourage cell-cell interactions than direct cell-substrate interactions. Thus, the impedance spectroscopy coupled with the proposed theoretical framework for PS substrates can sense and quantify the cellular interactions.

  18. Real-time sensing of epithelial cell-cell and cell-substrate interactions by impedance spectroscopy on porous substrates

    International Nuclear Information System (INIS)

    Mondal, D.; RoyChaudhuri, C.; Pal, D.

    2015-01-01

    Oxidized porous silicon (PS) is a common topographical biocompatible substrate that potentially provides a distinct in vitro environment for better understanding of in vivo behavior. But in the reported studies on oxidized PS, cell-cell and cell-substrate interactions have been detected only by fluorescent labeling. This paper is the first attempt to investigate real-time sensing of these interactions on HaCaT cells by label-free impedance spectroscopy on oxidized PS of two pore diameters (50 and 500 nm). One of the major requirements for successful impedance spectroscopy measurement is to restrict the channeling of electric field lines through the pores. To satisfy this criterion, we have designed the pore depths after analyzing the penetration of the medium by using computational fluid dynamics simulation. A distributed electrical model was also developed for estimating the various cellular attributes by considering a pseudorandom distribution of pores. It is observed from the impedance measurements and from the model that the proliferation rate increases for 50 nm pores but decreases for 500 nm pores compared to that for planar substrates. The rate of decrease in cell substrate separation (h) in the initial stage is more than the rate of increase in cell-cell junction resistance (R b ) corresponding to the initial adhesion phase of cells. It is observed that R b and h are higher for 50 nm pores than those for planar substrates, corresponding to the fact that substrates more conducive toward cell adhesion encourage cell-cell interactions than direct cell-substrate interactions. Thus, the impedance spectroscopy coupled with the proposed theoretical framework for PS substrates can sense and quantify the cellular interactions

  19. Quality improvement of organic thin films deposited on vibrating substrates

    Energy Technology Data Exchange (ETDEWEB)

    Paredes, Y.A.; Caldas, P.G.; Prioli, R.; Cremona, M., E-mail: cremona@fis.puc-rio.br

    2011-12-30

    Most of the Organic Light-Emitting Diodes (OLEDs) have a multilayered structure composed of functional organic layers sandwiched between two electrodes. Thin films of small molecules are generally deposited by thermal evaporation onto glass or other rigid or flexible substrates. The interface state between two organic layers in OLED device depends on the surface morphology of the layers and affects deeply the OLED performance. The morphology of organic thin films depends mostly on substrate temperature and deposition rate. Generally, the control of the substrate temperature allows improving the quality of the deposited films. For organic compounds substrate temperature cannot be increased too much due to their poor thermal stability. However, studies in inorganic thin films indicate that it is possible to modify the morphology of a film by using substrate vibration without increasing the substrate temperature. In this work, the effect of the resonance vibration of glass and silicon substrates during thermal deposition in high vacuum environment of tris(8-quinolinolate)aluminum(III) (Alq{sub 3}) and N,N Prime -Bis(naphthalene-2-yl)-N,N Prime -bis(phenyl)-benzidine ({beta}-NPB) organic thin films with different deposition rates was investigated. The vibration used was in the range of hundreds of Hz and the substrates were kept at room temperature during the process. The nucleation and subsequent growth of the organic films on the substrates have been studied by atomic force microscopy technique. For Alq{sub 3} and {beta}-NPB films grown with 0.1 nm/s as deposition rate and using a frequency of 100 Hz with oscillation amplitude of some micrometers, the results indicate a reduction of cluster density and a roughness decreasing. Moreover, OLEDs fabricated with organic films deposited under these conditions improved their power efficiency, driven at 4 mA/cm{sup 2}, passing from 0.11 lm/W to 0.24 lm/W with an increase in their luminance of about 352 cd/m{sup 2

  20. Interactions between Casein kinase Iepsilon (CKIepsilon and two substrates from disparate signaling pathways reveal mechanisms for substrate-kinase specificity.

    Directory of Open Access Journals (Sweden)

    Caroline Lund Dahlberg

    Full Text Available Members of the Casein Kinase I (CKI family of serine/threonine kinases regulate diverse biological pathways. The seven mammalian CKI isoforms contain a highly conserved kinase domain and divergent amino- and carboxy-termini. Although they share a preferred target recognition sequence and have overlapping expression patterns, individual isoforms often have specific substrates. In an effort to determine how substrates recognize differences between CKI isoforms, we have examined the interaction between CKIepsilon and two substrates from different signaling pathways.CKIepsilon, but not CKIalpha, binds to and phosphorylates two proteins: Period, a transcriptional regulator of the circadian rhythms pathway, and Disheveled, an activator of the planar cell polarity pathway. We use GST-pull-down assays data to show that two key residues in CKIalpha's kinase domain prevent Disheveled and Period from binding. We also show that the unique C-terminus of CKIepsilon does not determine Dishevelled's and Period's preference for CKIepsilon nor is it essential for binding, but instead plays an auxillary role in stabilizing the interactions of CKIepsilon with its substrates. We demonstrate that autophosphorylation of CKIepsilon's C-terminal tail prevents substrate binding, and use mass spectrometry and chemical crosslinking to reveal how a phosphorylation-dependent interaction between the C-terminal tail and the kinase domain prevents substrate phosphorylation and binding.The biochemical interactions between CKIepsilon and Disheveled, Period, and its own C-terminus lead to models that explain CKIepsilon's specificity and regulation.

  1. Design, fabrication and characterisation of advanced substrate crosstalk suppression structures in silicon on insulator substrates with buried ground planes (GPSOI)

    International Nuclear Information System (INIS)

    Stefanou, Stefanos

    2002-07-01

    Substrate crosstalk or coupling has been acknowledged to be a limiting factor in mixed signal RF integration. Although high levels of integration and high frequencies of operation are desirable for mixed mode RF and microwave circuits, they make substrate crosstalk more pronounced and may lead to circuit performance degradation. High signal isolation is dictated by requirements for low power dissipation, reduced number of components and lower integration costs for feasible system-on-chip (SoC) solutions. Substrate crosstalk suppression in ground plane silicon-on-insulator (GPSOI) substrates is investigated in this thesis. Test structures are designed and fabricated on SOI substrates with a buried WSi 2 plane that is connected to ground; hence it is called a ground plane. A Faraday cage structure that exhibits very high degrees of signal isolation is presented and compared to other SOI isolation schemes. The Faraday cage structure is shown to achieve 20 dB increased isolation in the frequency range of 0.5-50 GHz compared to published data for high resistivity (200 Ωcm) thin film SOI substrates with no ground planes, but where capacitive guard rings were used. The measurement results are analysed with the aid of planar electromagnetic simulators and compact lumped element models of all the fabricated test structures are developed. The accuracy of the lumped models is validated against experimental measurements. (author)

  2. Optimization of the enzyme system for hydrolysis of pretreated lignocellulose substrates; Optimering av enzymsystemet foer hydrolys av foerbehandlade lignocellulosa substrat

    Energy Technology Data Exchange (ETDEWEB)

    Tjerneld, Folke [Lund univ., (Sweden). Dept. of Biochemistry

    2000-06-01

    This project aims to clarify the reasons for the slow and incomplete enzymatic hydrolysis of certain lignocellulose substrates, particularly softwood e.g. spruce. Based on this knowledge we will optimize the enzyme system so that the yield of fermentable sugars is increased as well as the rate of hydrolysis. We will also study methods for recycling of the enzymes in the process by adsorption on fresh substrate. Progress in these areas will lead to improved process economy in an ethanol process. We collaborate with Chemical Engineering on hydrolysis of pretreated lignocellulose substrates and with Analytical Chemistry and Applied Microbiology on analysis of potential inhibitors. Within this main research direction the work at Biochemistry during this project period (since 970701) has been focused on the following areas: (1) Studies of the role of substrate properties in the enzymatic hydrolysis to clarify the reasons for the decrease in the rate of hydrolysis; (2) enzyme adsorption on lignin; (3) studies of recently identified low molecular weight endo glucanases which may be used for more effective penetration of small pores in pretreated substrates (this part is financed by the Nordic Energy Research Program). Central results during the period: In order to study the role of substrate properties for hydrolysis we have initiated investigations on steam pretreated substrates with several techniques. Measurements of pore sizes have been done with probe molecules of known molecular weights. Results show that probe molecules with diameters larger than 50 Aangstroem can more easily penetrate pretreated willow compared with spruce, which can be a part of the explanation for the better hydrolysability of hardwood substrates compared with softwood. We have started studies with electron microscopy of pretreated substrates at different degrees of enzymatic hydrolysis. With scanning electron microscopy (SEM) we can see significant differences in substrate structure in

  3. Tribology of bio-inspired nanowrinkled films on ultrasoft substrates.

    Science.gov (United States)

    Lackner, Juergen M; Waldhauser, Wolfgang; Major, Lukasz; Teichert, Christian; Hartmann, Paul

    2013-01-01

    Biomimetic design of new materials uses nature as antetype, learning from billions of years of evolution. This work emphasizes the mechanical and tribological properties of skin, combining both hardness and wear resistance of its surface (the stratum corneum) with high elasticity of the bulk (epidermis, dermis, hypodermis). The key for combination of such opposite properties is wrinkling, being consequence of intrinsic stresses in the bulk (soft tissue): Tribological contact to counterparts below the stress threshold for tissue trauma occurs on the thick hard stratum corneum layer pads, while tensile loads smooth out wrinkles in between these pads. Similar mechanism offers high tribological resistance to hard films on soft, flexible polymers, which is shown for diamond-like carbon (DLC) and titanium nitride thin films on ultrasoft polyurethane and harder polycarbonate substrates. The choice of these two compared substrate materials will show that ultra-soft substrate materials are decisive for the distinct tribological material. Hierarchical wrinkled structures of films on these substrates are due to high intrinsic compressive stress, which evolves during high energetic film growth. Incremental relaxation of these stresses occurs by compound deformation of film and elastic substrate surface, appearing in hierarchical nano-wrinkles. Nano-wrinkled topographies enable high elastic deformability of thin hard films, while overstressing results in zigzag film fracture along larger hierarchical wrinkle structures. Tribologically, these fracture mechanisms are highly important for ploughing and sliding of sharp and flat counterparts on hard-coated ultra-soft substrates like polyurethane. Concentration of polyurethane deformation under the applied normal loads occurs below these zigzag cracks. Unloading closes these cracks again. Even cyclic testing do not lead to film delamination and retain low friction behavior, if the adhesion to the substrate is high and the initial

  4. TRIBOLOGY OF BIO-INSPIRED NANOWRINKLED FILMS ON ULTRASOFT SUBSTRATES

    Directory of Open Access Journals (Sweden)

    Juergen M. Lackner

    2013-03-01

    Full Text Available Biomimetic design of new materials uses nature as antetype, learning from billions of years of evolution. This work emphasizes the mechanical and tribological properties of skin, combining both hardness and wear resistance of its surface (the stratum corneum with high elasticity of the bulk (epidermis, dermis, hypodermis. The key for combination of such opposite properties is wrinkling, being consequence of intrinsic stresses in the bulk (soft tissue: Tribological contact to counterparts below the stress threshold for tissue trauma occurs on the thick hard stratum corneum layer pads, while tensile loads smooth out wrinkles in between these pads. Similar mechanism offers high tribological resistance to hard films on soft, flexible polymers, which is shown for diamond-like carbon (DLC and titanium nitride thin films on ultrasoft polyurethane and harder polycarbonate substrates. The choice of these two compared substrate materials will show that ultra-soft substrate materials are decisive for the distinct tribological material. Hierarchical wrinkled structures of films on these substrates are due to high intrinsic compressive stress, which evolves during high energetic film growth. Incremental relaxation of these stresses occurs by compound deformation of film and elastic substrate surface, appearing in hierarchical nano-wrinkles. Nano-wrinkled topographies enable high elastic deformability of thin hard films, while overstressing results in zigzag film fracture along larger hierarchical wrinkle structures. Tribologically, these fracture mechanisms are highly important for ploughing and sliding of sharp and flat counterparts on hard-coated ultra-soft substrates like polyurethane. Concentration of polyurethane deformation under the applied normal loads occurs below these zigzag cracks. Unloading closes these cracks again. Even cyclic testing do not lead to film delamination and retain low friction behavior, if the adhesion to the substrate is high

  5. Precise micropatterning of silver nanoparticles on plastic substrates

    Energy Technology Data Exchange (ETDEWEB)

    Ammosova, Lena; Jiang, Yu; Suvanto, Mika; Pakkanen, Tapani A., E-mail: tapani.pakkanen@uef.fi

    2017-04-15

    Highlights: • Silver ink has been deposited on plastic substrate and silver nanoparticles have been produced. • 3D control allows both ink superimposing and deposition on complicated surfaces. • Polyol method ensures the formation of metallic mircopatterns with high uniformity. • Substrate wettability, ink volume, and sintering temperature influences deposited patterns. - Abstract: Conventional fabrication methods to obtain metal patterns on polymer substrates are restricted by high operating temperature and complex preparation steps. The present study demonstrates a simple yet versatile method for preparation of silver nanoparticle micropatterns on polymer substrates with various surface geometry. With the microworking robot technique, we were able not only to directly structure the surface, but also precisely deposit silver nanoparticle ink on the desired surface location with the minimum usage of ink material. The prepared silver nanoparticle ink, containing silver cations and polyethylene glycol (PEG) as a reducing agent, yields silver nanoparticle micropatterns on plastic substrates at low sintering temperature without any contamination. The influence of the ink behaviour was studied, such as substrate wettability, ink volume, and sintering temperature. The ultraviolet visible (UV–vis), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) measurements revealed the formation of micropatterns with uniformly distributed silver nanoparticles. The prepared patterns are expected to have a broad range of applications in optics, medicine, and sensor devices owing to the unique properties of silver. Furthermore, the deposition of a chemical compound, which is different from the substrate material, not only adds a fourth dimension to the prestructured three-dimensional (3D) surfaces, but also opens new application areas to the conventional surface structures.

  6. Precise micropatterning of silver nanoparticles on plastic substrates

    International Nuclear Information System (INIS)

    Ammosova, Lena; Jiang, Yu; Suvanto, Mika; Pakkanen, Tapani A.

    2017-01-01

    Highlights: • Silver ink has been deposited on plastic substrate and silver nanoparticles have been produced. • 3D control allows both ink superimposing and deposition on complicated surfaces. • Polyol method ensures the formation of metallic mircopatterns with high uniformity. • Substrate wettability, ink volume, and sintering temperature influences deposited patterns. - Abstract: Conventional fabrication methods to obtain metal patterns on polymer substrates are restricted by high operating temperature and complex preparation steps. The present study demonstrates a simple yet versatile method for preparation of silver nanoparticle micropatterns on polymer substrates with various surface geometry. With the microworking robot technique, we were able not only to directly structure the surface, but also precisely deposit silver nanoparticle ink on the desired surface location with the minimum usage of ink material. The prepared silver nanoparticle ink, containing silver cations and polyethylene glycol (PEG) as a reducing agent, yields silver nanoparticle micropatterns on plastic substrates at low sintering temperature without any contamination. The influence of the ink behaviour was studied, such as substrate wettability, ink volume, and sintering temperature. The ultraviolet visible (UV–vis), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) measurements revealed the formation of micropatterns with uniformly distributed silver nanoparticles. The prepared patterns are expected to have a broad range of applications in optics, medicine, and sensor devices owing to the unique properties of silver. Furthermore, the deposition of a chemical compound, which is different from the substrate material, not only adds a fourth dimension to the prestructured three-dimensional (3D) surfaces, but also opens new application areas to the conventional surface structures.

  7. HIV-1 protease-substrate coevolution in nelfinavir resistance.

    Science.gov (United States)

    Kolli, Madhavi; Ozen, Ayşegül; Kurt-Yilmaz, Nese; Schiffer, Celia A

    2014-07-01

    Resistance to various human immunodeficiency virus type 1 (HIV-1) protease inhibitors (PIs) challenges the effectiveness of therapies in treating HIV-1-infected individuals and AIDS patients. The virus accumulates mutations within the protease (PR) that render the PIs less potent. Occasionally, Gag sequences also coevolve with mutations at PR cleavage sites contributing to drug resistance. In this study, we investigated the structural basis of coevolution of the p1-p6 cleavage site with the nelfinavir (NFV) resistance D30N/N88D protease mutations by determining crystal structures of wild-type and NFV-resistant HIV-1 protease in complex with p1-p6 substrate peptide variants with L449F and/or S451N. Alterations of residue 30's interaction with the substrate are compensated by the coevolving L449F and S451N cleavage site mutations. This interdependency in the PR-p1-p6 interactions enhances intermolecular contacts and reinforces the overall fit of the substrate within the substrate envelope, likely enabling coevolution to sustain substrate recognition and cleavage in the presence of PR resistance mutations. Resistance to human immunodeficiency virus type 1 (HIV-1) protease inhibitors challenges the effectiveness of therapies in treating HIV-1-infected individuals and AIDS patients. Mutations in HIV-1 protease selected under the pressure of protease inhibitors render the inhibitors less potent. Occasionally, Gag sequences also mutate and coevolve with protease, contributing to maintenance of viral fitness and to drug resistance. In this study, we investigated the structural basis of coevolution at the Gag p1-p6 cleavage site with the nelfinavir (NFV) resistance D30N/N88D protease mutations. Our structural analysis reveals the interdependency of protease-substrate interactions and how coevolution may restore substrate recognition and cleavage in the presence of protease drug resistance mutations. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  8. Stability of perovskite solar cells on flexible substrates

    Science.gov (United States)

    Tam, Ho Won; Chen, Wei; Liu, Fangzhou; He, Yanling; Leung, Tik Lun; Wang, Yushu; Wong, Man Kwong; Djurišić, Aleksandra B.; Ng, Alan Man Ching; He, Zhubing; Chan, Wai Kin; Tang, Jinyao

    2018-02-01

    Perovskite solar cells are emerging photovoltaic technology with potential for low cost, high efficiency devices. Currently, flexible devices efficiencies over 15% have been achieved. Flexible devices are of significant interest for achieving very low production cost via roll-to-roll processing. However, the stability of perovskite devices remains a significant challenge. Unlike glass substrate which has negligible water vapor transmission rate (WVTR), polymeric flexible film substrates suffer from high moisture permeability. As PET and PEN flexible substrates exhibit higher water permeability then glass, transparent flexible backside encapsulation should be used to maximize light harvesting in perovskite layer while WVTR should be low enough. Wide band gap materials are transparent in the visible spectral range low temperature processable and can be a moisture barrier. For flexible substrates, approaches like atomic layer deposition (ALD) and low temperature solution processing could be used for metal oxide deposition. In this work, ALD SnO2, TiO2, Al2O3 and solution processed spin-on-glass was used as the barrier layer on the polymeric side of indium tin oxide (ITO) coated PEN substrates. The UV-Vis transmission spectra of the prepared substrates were investigated. Perovskite solar cells will be fabricated and stability of the devices were encapsulated with copolymer films on the top side and tested under standard ISOS-L-1 protocol and then compared to the commercial unmodified ITO/PET or ITO/PEN substrates. In addition, devices with copolymer films laminated on both sides successfully surviving more than 300 hours upon continuous AM1.5G illumination were demonstrated.

  9. Light alloys as substrate material for bipolar plates; Leichtmetall-Legierungen als Substrat fuer Bipolarplatten

    Energy Technology Data Exchange (ETDEWEB)

    Schicke, R. [PSFU GmbH, Wernigerode (Germany)

    2008-07-01

    Light alloys as substrate material for bipolar plates in fuel cells offer a number of advantages compared to stainless steel sheets. First, the specific weight is smaller, costs are lower, but also bulk properties like thermal and electric conductivities are much better than in the case of stainless steel. Regarding graphite polymer composite materials, the electric conductivity of light alloys again is much higher leading to a considerably lower internal resistance of the cells. Metal sheets, in general, are more attractive with respect to building up compact stacks with high power densities since metal sheets can be produced easily down to thicknesses of around 0.1 mm, whereby graphite composite materials most often have a thickness of at least around 2 mm. In addition, the economics of using light alloys as bipolar plate material is advantageous also for small and medium quantities of production (for instance making use of photochemical etching), but also for high volume production where both conventional techniques like stamping and also more advanced processes like hydroforming can be employed. A major challenge is the identification and technological control and improvement of surface modification / coating processes which lead to low ohmic contact resistances and a good corrosion protection under the electrochemical conditions within a fuel cell environment. Different coating technologies and the characteristics of several coatings will be discussed. (orig.)

  10. Hot Films on Ceramic Substrates for Measuring Skin Friction

    Science.gov (United States)

    Noffz, Greg; Leiser, Daniel; Bartlett, Jim; Lavine, Adrienne

    2003-01-01

    Hot-film sensors, consisting of a metallic film on an electrically nonconductive substrate, have been used to measure skin friction as far back as 1931. A hot film is maintained at an elevated temperature relative to the local flow by passing an electrical current through it. The power required to maintain the specified temperature depends on the rate at which heat is transferred to the flow. The heat transfer rate correlates to the velocity gradient at the surface, and hence, with skin friction. The hot-film skin friction measurement method is most thoroughly developed for steady-state conditions, but additional issues arise under transient conditions. Fabricating hot-film substrates using low-thermal-conductivity ceramics can offer advantages over traditional quartz or polyester-film substrates. First, a low conductivity substrate increases the fraction of heat convected away by the fluid, thus increasing sensitivity to changes in flow conditions. Furthermore, the two-part, composite nature of the substrate allows the installation of thermocouple junctions just below the hot film, which can provide an estimate of the conduction heat loss.

  11. The influence of substrate material on ascidian larval settlement.

    Science.gov (United States)

    Chase, Anna L; Dijkstra, Jennifer A; Harris, Larry G

    2016-05-15

    Submerged man-made structures present novel habitat for marine organisms and often host communities that differ from those on natural substrates. Although many factors are known to contribute to these differences, few studies have directly examined the influence of substrate material on organism settlement. We quantified larval substrate preferences of two species of ascidians, Ciona intestinalis (cryptogenic, formerly C. intestinalis type B) and Botrylloides violaceus (non-native), on commonly occurring natural (granite) and man-made (concrete, high-density polyethylene, PVC) marine materials in laboratory trials. Larvae exhibited species-specific settlement preferences, but generally settled more often than expected by chance on concrete and HDPE. Variation in settlement between materials may reflect preferences for rougher substrates, or may result from the influence of leached chemicals on ascidian settlement. These findings indicate that an experimental plate material can influence larval behavior and may help us understand how substrate features may contribute to differences in settlement in the field. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Infrared photothermal imaging of trace explosives on relevant substrates

    Science.gov (United States)

    Kendziora, Christopher A.; Furstenberg, Robert; Papantonakis, Michael; Nguyen, Viet; Borchert, James; Byers, Jeff; McGill, R. Andrew

    2013-06-01

    We are developing a technique for the stand-off detection of trace explosives on relevant substrate surfaces using photo-thermal infrared (IR) imaging spectroscopy (PT-IRIS). This approach leverages one or more compact IR quantum cascade lasers, tuned to strong absorption bands in the analytes and directed to illuminate an area on a surface of interest. An IR focal plane array is used to image the surface and detect small increases in thermal emission upon laser illumination. The PT-IRIS signal is processed as a hyperspectral image cube comprised of spatial, spectral and temporal dimensions as vectors within a detection algorithm. The ability to detect trace analytes on relevant substrates is critical for stand-off applications, but is complicated by the optical and thermal analyte/substrate interactions. This manuscript describes recent PT-IRIS experimental results and analysis for traces of RDX, TNT, ammonium nitrate (AN) and sucrose on relevant substrates (steel, polyethylene, glass and painted steel panels). We demonstrate that these analytes can be detected on these substrates at relevant surface mass loadings (10 μg/cm2 to 100 μg/cm2) even at the single pixel level.

  13. Substrate considerations for graphene synthesis on thin copper films

    International Nuclear Information System (INIS)

    Howsare, Casey A; Robinson, Joshua A; Weng Xiaojun; Bojan, Vince; Snyder, David

    2012-01-01

    Chemical vapor deposition on copper substrates is a primary technique for synthesis of high quality graphene films over large areas. While well-developed processes are in place for catalytic growth of graphene on bulk copper substrates, chemical vapor deposition of graphene on thin films could provide a means for simplified device processing through the elimination of the layer transfer process. Recently, it was demonstrated that transfer-free growth and processing is possible on SiO 2 . However, the Cu/SiO 2 /Si material system must be stable at high temperatures for high quality transfer-free graphene. This study identifies the presence of interdiffusion at the Cu/SiO 2 interface and investigates the influence of metal (Ni, Cr, W) and insulating (Si 3 N 4 , Al 2 O 3 , HfO 2 ) diffusion barrier layers on Cu–SiO 2 interdiffusion, as well as graphene structural quality. Regardless of barrier choice, we find the presence of Cu diffusion into the silicon substrate as well as the presence of Cu–Si–O domains on the surface of the copper film. As a result, we investigate the choice of a sapphire substrate and present evidence that it is a robust substrate for synthesis and processing of high quality, transfer-free graphene. (paper)

  14. Adhesion mechanisms of nanoparticle silver to substrate materials: identification

    International Nuclear Information System (INIS)

    Joo, Sungchul; Baldwin, Daniel F

    2010-01-01

    Nanoparticle silver (NPS) conductors are increasingly being investigated for printed electronics applications. However, the adhesion mechanism of the nanoparticle silver to substrate materials has not been identified yet. In particular, the adhesion of NPS to organic materials such as the widely used polyimide Kapton HN and Kapton FPC dry films is concerned with low adhesion strength because the processed polymer surface is chemically inert. Moreover, its adhesion to substrate materials such as benzocyclobutene (BCB), copper and aluminum was found to be very weak. Therefore, in this paper, the mechanisms of NPS adhesion to organic and inorganic materials are identified as the first step in improving NPS adhesion strength. Improving the adhesion strength of NPS will be the key issue for printed electronics applications. The adhesion of NPS to substrate materials was found to be mainly attributed to van der Waals forces based on particle adhesion mechanisms. This finding provides the initiative of developing an adhesion prediction model of NPS to substrate materials in order to provide guidelines for improving the NPS adhesion strength to the substrate materials used in printed electronics.

  15. Responses of low pressure Andersen sampler for collecting substrates

    Energy Technology Data Exchange (ETDEWEB)

    Yamasaki, K. [Research Reactor Institute, Kyoto University, Osaka (Japan); Yamada, Y.; Miyamoto, K.; Shimo, M. [Division of Radiotoxicology and Protection, National Institute of Radiological Sciences, Chiba (Japan)

    2000-05-01

    Some types of low pressure cascade impactors (Andersen, Berner, Davies, and MOUDI etc) have been used to measure the activity size distribution of radon progeny in the environment. In spite of their careful uses, their nonideal responses are not adequately known. Some important factors such as the wall loss, electrostatic attraction, and the surface nature of collecting substrates may affect the reliability of the impactor data. Size selective characteristics of a low pressure Andersen sampler for various collecting substrates were examined in a radon exposure chamber using several kinds of liquid (DOS) or solid (carnauba wax) carrier aerosols labelled with radon progeny. These carrier aerosols were produced by commercial condensation aerosol generator. Tested collecting substrates were, (1) uncoated clean stainless steel plate, (2) silicone oil or grease coated stainless steel plate, (3) polyethylene sheet covered stainless steel plate, (4) membrane filter, (5) glass fiber filter, and (6) quartz fiber filter. In the case of collecting liquid or sticky carrier aerosols, the effect of particle bounce was small and nearly any substrates might used on the impaction plate. On the other hand, in the case of solid carrier aerosols, an adhesive layer such as grease or oil might have to be applied to the substrate. It was concluded that a low pressure cascade impactor such as Andersen sampler might need an appropriate calibration procedure including the interstage characteristics for determining the accurate activity size distribution. (author)

  16. The substrate matters in the Raman spectroscopy analysis of cells

    Science.gov (United States)

    Mikoliunaite, Lina; Rodriguez, Raul D.; Sheremet, Evgeniya; Kolchuzhin, Vladimir; Mehner, Jan; Ramanavicius, Arunas; Zahn, Dietrich R. T.

    2015-08-01

    Raman spectroscopy is a powerful analytical method that allows deposited and/or immobilized cells to be evaluated without complex sample preparation or labeling. However, a main limitation of Raman spectroscopy in cell analysis is the extremely weak Raman intensity that results in low signal to noise ratios. Therefore, it is important to seize any opportunity that increases the intensity of the Raman signal and to understand whether and how the signal enhancement changes with respect to the substrate used. Our experimental results show clear differences in the spectroscopic response from cells on different surfaces. This result is partly due to the difference in spatial distribution of electric field at the substrate/cell interface as shown by numerical simulations. We found that the substrate also changes the spatial location of maximum field enhancement around the cells. Moreover, beyond conventional flat surfaces, we introduce an efficient nanostructured silver substrate that largely enhances the Raman signal intensity from a single yeast cell. This work contributes to the field of vibrational spectroscopy analysis by providing a fresh look at the significance of the substrate for Raman investigations in cell research.

  17. Gecko Adhesion on Wet and Dry Patterned Substrates.

    Directory of Open Access Journals (Sweden)

    Alyssa Y Stark

    Full Text Available Perhaps one of the most astounding characteristics of the gecko adhesive system is its versatility. Geckos can locomote across complex substrates in a variety of conditions with apparent ease. In contrast, many of our synthetic pressure sensitive adhesives fail on substrates that are dirty, wet or rough. Although many studies have investigated the effect of environmental challenges on performance, the interaction of multiple, potentially compromising variables is studied less often. Here we focus on substrate structure and surface water, both of which are highly relevant to the biological system and to synthetic design. To do this we utilized a highly controlled, patterned substrate (Sharklet®, by Sharklet® Technologies Inc.. This allowed us to test independently and jointly the effects of reduced surface area substrates, with a defined pattern, on adhesion in both air and water. Our results show that adhesion is not significantly impaired in air, whereas surface area and pattern significantly affect adhesion in water. These findings highlight the need to study multiple parameters that are relevant to the gecko adhesive system to further improve our understanding of the biological system and to design better, more versatile synthetics.

  18. Cultivation of Schizophyllum commune mushroom on different wood substrates

    Directory of Open Access Journals (Sweden)

    P.N. Dasanayaka

    2017-07-01

    Full Text Available Schizophyllum commune is an edible mushroom grown on wood under natural conditions. Present study focused on cultivation of S.commune on different wood substrates since it is not commercially cultivated. A pure culture of S. commune was obtained by growing a tissue of the mushroom on Potato Dextrose Agar (PDA medium. Spawns were produced by growing the mycelium on paddy grains. Mushroom was cultivated on sawdust of seven different wood substrates. The maximum yield was observed in sawdust of jackfruit (Artocarpusheterophyllus followed by sawdust of rambutan (Nepheliumlappaceum and country almond (Terminaliacatappa. A significant difference was not observed when mango (Mangiferaindica elephant apple (Dilleniaindica, tulip wood tree (Harpulliaarborea and thungfaa (Alstoniamacrophylla sawdust used as substrate. The lowest yield was observed in thungfaa (Alstoniamacrophylla sawdust. Effect of some additives on the yield was studied and significant difference in yield was observed when rice bran and used-tea leaves used as additives. Effect of rice bran on yield was studied using different ratios of sawdust to rice bran and the highest was observed in 2:1 ratio of sawdust to rice bran. The best incubating temperature for mycelial growth on the substrate was 350C. The composition of the mushroom on a dry weight basis was; 71.4% moisture, 23.35% crude protein and 6% ash. Tested wood species are promising substrates for cultivation of S.communeas cottage industry.

  19. Effect Of Various Substrates On Eudrilus eugeniae (Oligochaeta Production

    Directory of Open Access Journals (Sweden)

    Mpoame, M.

    1994-01-01

    Full Text Available In order to assess the effects of substrate on Eudrilus eugeniae production, 360 worms were raised in plastic buckets containing manure either from cattle (BV, from sheep (CM, from rabbit (CL, and from chicken (FP. At the end of the experiment which lasted 4 months, worm productivity was estimated at 111.9 g/kg of substrate in CL, 86.3 g/kg in CM, and 33.0 g/kg in BV. Substrate FP was inadequate as worms escaped from it. In another trial aiming at determining some of E. Eugeniae's reproductive parameters, each of the 3 substrates BV, CL, and CM was distributed into 15 plastic half bottles and was inoculated with one cocoon. In the 3 substrates cocoon incubation time and growing period were estimated at 3 to 4 weeks and 6 to 7 weeks respectively. Cocoons were laid approximately a week after the pairing of worms. On the average, each worm laid 2 cocoons per week.

  20. Chemical vapor deposition of yttria stabilized zirconia in porous substrates

    International Nuclear Information System (INIS)

    Carolan, M.F.; Michaels, J.N.

    1987-01-01

    Electrochemical vapor deposition (EVD) of yttria stabilized zirconia (YSZ) is the preferred route to the production of thin films of YSZ on porous substrates. This process has been used in the construction of both fuel cells and steam electrolyzers. A critical aspect of the EVD process is an initial chemical vapor deposition phase in which the pores of a porous substrate are plugged by YSZ. In this process, water vapor and a mixture of gaseous zirconium chloride and yttrium chloride diffuse into the porous substrate from opposite sides and react to form YSZ and HCl ga. During the second stage of the process a continuous dense film of electrolyte is formed by a tarnishing-type process. Experimentally it is observed that the pores plug within a few pore diameters of the metal chloride face of the substrate. A kinetic rate expression that is first order in metal chloride but zero order in water is best able to explain this phenomenon. With this rate expression, the pores always plug near the metal chloride face. The model predicts less pore narrowing to occur as the ratio of the reaction rate to the diffusion rate of the metal chloride is increased. A kinetic rate expression that is first order in both water and metal chloride predicts that the pores plug much deeper in the substrate

  1. Weather resistance of inkjet prints on plastic substrates

    Directory of Open Access Journals (Sweden)

    Rozália Szentgyörgyvölgyi

    2015-06-01

    Full Text Available The development of wide format inkjet printers made the technology available for large area commercials. Outdoor advertising uses a wide range of substrate including paperboard, vinyl, canvas, mesh; the material of the substrate itself has to endure the physical and chemical effects of local weather. Weather elements (humidity, wind, solar irradiation degrade printed products inevitably; plastic products have better resistance against them, than paper based substrates. Service life of the printed product for outdoor application is a key parameter from the customer’s point of view. There are two ways to estimate expected lifetime: on site outdoor testing or laboratory testing. In both cases weathering parameters can be monitored, however laboratory testing devices may produce the desired environmental effects and thus accelerate the aging process. Our research objective was to evaluate the effects of artificial weathering on prints produced by inkjet technology on plastic substrates. We used a large format CMYK inkjet printer (Mutoh Rockhopper II, with Epson DX 4 print heads to print our test chart on two similar substrates (PVC coated tarpaulins with grammages 400 g/m2 and 440 g/m2. Specimen were aged in an Atlas Suntest XLS+ material tester device for equal time intervals. We measured and calculated the gradual changes of the optical properties (optical density, tone value, colour shifts of the test prints.

  2. A liquid aluminum corrosion resistance surface on steel substrate

    International Nuclear Information System (INIS)

    Wang Deqing; Shi Ziyuan; Zou Longjiang

    2003-01-01

    The process of hot dipping pure aluminum on a steel substrate followed by oxidation was studied to form a surface layer of aluminum oxide resistant to the corrosion of aluminum melt. The thickness of the pure aluminum layer on the steel substrate is reduced with the increase in temperature and time in initial aluminizing, and the thickness of the aluminum layer does not increase with time at given temperature when identical temperature and complete wetting occur between liquid aluminum and the substrate surface. The thickness of the Fe-Al intermetallic layer on the steel base is increased with increasing bath temperature and time. Based on the experimental data and the mathematics model developed by the study, a maximum exists in the thickness of the Fe-Al intermetallic at certain dipping temperature. X-ray diffraction (XRD) and energy dispersive X-ray (EDX) analysis reveals that the top portion of the steel substrate is composed of a thin layer of α-Al 2 O 3 , followed by a thinner layer of FeAl 3 , and then a much thicker one of Fe 2 Al 5 on the steel base side. In addition, there is a carbon enrichment zone in diffusion front. The aluminum oxide surface formed on the steel substrate is in perfect condition after corrosion test in liquid aluminum at 750 deg. C for 240 h, showing extremely good resistance to aluminum melt corrosion

  3. Computational Approach for Epitaxial Polymorph Stabilization through Substrate Selection

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Hong; Dwaraknath, Shyam S.; Garten, Lauren; Ndione, Paul; Ginley, David; Persson, Kristin A.

    2016-05-25

    With the ultimate goal of finding new polymorphs through targeted synthesis conditions and techniques, we outline a computational framework to select optimal substrates for epitaxial growth using first principle calculations of formation energies, elastic strain energy, and topological information. To demonstrate the approach, we study the stabilization of metastable VO2 compounds which provides a rich chemical and structural polymorph space. We find that common polymorph statistics, lattice matching, and energy above hull considerations recommends homostructural growth on TiO2 substrates, where the VO2 brookite phase would be preferentially grown on the a-c TiO2 brookite plane while the columbite and anatase structures favor the a-b plane on the respective TiO2 phases. Overall, we find that a model which incorporates a geometric unit cell area matching between the substrate and the target film as well as the resulting strain energy density of the film provide qualitative agreement with experimental observations for the heterostructural growth of known VO2 polymorphs: rutile, A and B phases. The minimal interfacial geometry matching and estimated strain energy criteria provide several suggestions for substrates and substrate-film orientations for the heterostructural growth of the hitherto hypothetical anatase, brookite, and columbite polymorphs. These criteria serve as a preliminary guidance for the experimental efforts stabilizing new materials and/or polymorphs through epitaxy. The current screening algorithm is being integrated within the Materials Project online framework and data and hence publicly available.

  4. Yield of lettuce grown in aquaponic system using different substrates

    Directory of Open Access Journals (Sweden)

    Rodrigo A. Jordan

    Full Text Available ABSTRACT In the aquaponic system, the characteristics of the materials used as substrate directly affect plant development, because besides acting as a support base, they must present a surface to fix microorganisms, responsible for the conversion of nutrients into forms more easily available to plants. Thus, the objective of this study was to evaluate the effect of four growing substrates on the yield of lettuce grown in aquaponic system. The experimental design was randomized blocks with four treatments, which corresponded to the substrates, and six replicates. Plants were grown using the nutrient film technique (NFT system. The substrates used in the experiment were: coconut shell fiber with crushed stone #3, expanded vermiculite, zeolite and phenolic foam. The treatment with phenolic foam was considered as the least suitable for lettuce cultivation in aquaponic system, because it caused lower yield (20.8 t ha-1. The treatment using coconut shell fiber with crushed stone #3 was considered as the most adequate, since it led to higher yield (39.9 t ha-1 compared with the other substrates analyzed.

  5. SiC substrate defects and III-N heteroepitaxy

    Energy Technology Data Exchange (ETDEWEB)

    Poust, B D [Department of Materials Science and Engineering, University of California, Los Angeles, CA 90095 (United States); Koga, T S [Department of Materials Science and Engineering, University of California, Los Angeles, CA 90095 (United States); Sandhu, R [Department of Materials Science and Engineering, University of California, Los Angeles, CA 90095 (United States); Heying, B [Northrop Grumman Space Technology, Space and Electronics Group, Redondo Beach, CA 90278 (United States); Hsing, R [Northrop Grumman Space Technology, Space and Electronics Group, Redondo Beach, CA 90278 (United States); Wojtowicz, M [Northrop Grumman Space Technology, Space and Electronics Group, Redondo Beach, CA 90278 (United States); Khan, A [Department of Electrical Engineering, University of South Carolina, Columbia, SC (United States); Goorsky, M S [Department of Materials Science and Engineering, University of California, Los Angeles, CA 90095 (United States)

    2003-05-21

    This study addressed how defects in SiC substrates influence the crystallographic properties of AlGaN/GaN layers deposited by metallorganic vapour phase epitaxy and by molecular beam epitaxy. We employed double crystal reflection x-ray topography using symmetric (0008) and (00012) reflections with CuK{alpha} radiation ({lambda} = 1.54 A) to image dislocations, micropipes, and low angle boundaries in SiC substrates. Lattice strain near the core of a micropipe defect was estimated to be of the order of 10{sup -7}. The substrates investigated exhibited radial patterns of strain and, primarily, of tilt of the order of tens of arcsec. After deposition of the AlGaN and GaN layers, DCXRT images were generated from the substrate (0008) or (00012) and GaN epitaxial layer (0004) reflections. Full-width at half-maximum values ranging from {approx}100 to 300 arcsec were typical of the GaN reflections, while those of the 4H-SiC reflections were {approx}20-70 arcsec. Micropipes, tilt boundaries, and inclusions in the SiC were shown to produce structural defects in the GaN layers. A clear correlation between SiC substrate defects and GaN defects has been established.

  6. SiC substrate defects and III-N heteroepitaxy

    International Nuclear Information System (INIS)

    Poust, B D; Koga, T S; Sandhu, R; Heying, B; Hsing, R; Wojtowicz, M; Khan, A; Goorsky, M S

    2003-01-01

    This study addressed how defects in SiC substrates influence the crystallographic properties of AlGaN/GaN layers deposited by metallorganic vapour phase epitaxy and by molecular beam epitaxy. We employed double crystal reflection x-ray topography using symmetric (0008) and (00012) reflections with CuKα radiation (λ = 1.54 A) to image dislocations, micropipes, and low angle boundaries in SiC substrates. Lattice strain near the core of a micropipe defect was estimated to be of the order of 10 -7 . The substrates investigated exhibited radial patterns of strain and, primarily, of tilt of the order of tens of arcsec. After deposition of the AlGaN and GaN layers, DCXRT images were generated from the substrate (0008) or (00012) and GaN epitaxial layer (0004) reflections. Full-width at half-maximum values ranging from ∼100 to 300 arcsec were typical of the GaN reflections, while those of the 4H-SiC reflections were ∼20-70 arcsec. Micropipes, tilt boundaries, and inclusions in the SiC were shown to produce structural defects in the GaN layers. A clear correlation between SiC substrate defects and GaN defects has been established

  7. Molecular docking simulations provide insights in the substrate binding sites and possible substrates of the ABCC6 transporter.

    Directory of Open Access Journals (Sweden)

    Mohammad Jakir Hosen

    Full Text Available The human ATP-binding cassette family C member 6 (ABCC6 gene encodes an ABC transporter protein (ABCC6, primarily expressed in liver and kidney. Mutations in the ABCC6 gene cause pseudoxanthoma elasticum (PXE, an autosomal recessive connective tissue disease characterized by ectopic mineralization of the elastic fibers. The pathophysiology underlying PXE is incompletely understood, which can at least partly be explained by the undetermined nature of the ABCC6 substrates as well as the unknown substrate recognition and binding sites. Several compounds, including anionic glutathione conjugates (N-ethylmaleimide; NEM-GS and leukotriene C4 (LTC4 were shown to be modestly transported in vitro; conversely, vitamin K3 (VK3 was demonstrated not to be transported by ABCC6. To predict the possible substrate binding pockets of the ABCC6 transporter, we generated a 3D homology model of ABCC6 in both open and closed conformation, qualified for molecular docking and virtual screening approaches. By docking 10 reported in vitro substrates in our ABCC6 3D homology models, we were able to predict the substrate binding residues of ABCC6. Further, virtual screening of 4651 metabolites from the Human Serum Metabolome Database against our open conformation model disclosed possible substrates for ABCC6, which are mostly lipid and biliary secretion compounds, some of which are found to be involved in mineralization. Docking of these possible substrates in the closed conformation model also showed high affinity. Virtual screening expands this possibility to explore more compounds that can interact with ABCC6, and may aid in understanding the mechanisms leading to PXE.

  8. Thermal and Electrical Characterization of Alumina Substrate for Microelectronic Applications

    International Nuclear Information System (INIS)

    Ahmad, S.; Ibrahim, A.; Alias, R.; Shapee, S. M.; Ambak, Z.; Zakaria, S. Z.; Yahya, M. R.; Mat, A. F. A.

    2010-01-01

    This paper reports the effect of sintering temperature on thermal and electrical properties of alumina material as substrate for microelectronic devices. Alumina materials in the form of green sheet with 1 mm thickness were sintered at 1100 deg. C, 1300 deg. C and 1500 deg. C for about 20 hours using heating and cooling rates of 2 deg. C/min. The densities were measured using densitometer and the microstructures of the samples were analyzed using SEM micrographs. Meanwhile thermal and electrical properties of the samples were measured using flash method and impedance analyzer respectively. It was found that thermal conductivity and thermal diffusivity of the substrate increases as sintering temperature increases. It was found also that the dielectric constant of alumina substrate increases as the sintering temperature increases.

  9. Thermal and Electrical Characterization of Alumina Substrate for Microelectronic Applications

    Science.gov (United States)

    Ahmad, S.; Ibrahim, A.; Alias, R.; Shapee, S. M.; Ambak, Z.; Zakaria, S. Z.; Yahya, M. R.; Mat, A. F. A.

    2010-03-01

    This paper reports the effect of sintering temperature on thermal and electrical properties of alumina material as substrate for microelectronic devices. Alumina materials in the form of green sheet with 1 mm thickness were sintered at 1100° C, 1300° C and 1500° C for about 20 hours using heating and cooling rates of 2° C/min. The densities were measured using densitometer and the microstructures of the samples were analyzed using SEM micrographs. Meanwhile thermal and electrical properties of the samples were measured using flash method and impedance analyzer respectively. It was found that thermal conductivity and thermal diffusivity of the substrate increases as sintering temperature increases. It was found also that the dielectric constant of alumina substrate increases as the sintering temperature increases.

  10. Thin polymer films on chemically patterned, corrugated substrates

    International Nuclear Information System (INIS)

    Geoghegan, Mark; Wang Chun; Rehse, Nicolaus; Magerle, Robert; Krausch, Georg

    2005-01-01

    We study the effect of a chemical pattern on the wetting and dewetting behaviour of thin polystyrene (PS) films on regularly corrugated silicon substrates. Our results reveal that the film preparation, annealing method, and confinement play a critical role in the final film structure. On evaporating gold on both sides of the facets (such that it covered the crests of the facets, and not the troughs), we observed dewetting, which proceeded to the gold, demonstrating an enthalpic effect contrary to the outcome previously observed when gold was only evaporated on one side of the facet. We also coated the substrate with octadecyltrichlorosilane (OTS); this led to a gold and OTS striped structure. PS films several nanometres thick dewet such substrates, with a preferential direction for dewetting in the direction of the stripes forming droplets of a considerably larger size than the stripes

  11. Retrospective dosimetry with alumina substrate from electronic components

    International Nuclear Information System (INIS)

    Ekendahl, D.; Judas, L.

    2012-01-01

    Alumina substrate can be found in electronic components used in portable electronic devices. The material is radiation sensitive and can be applied in dosimetry using thermally or optically stimulated luminescence. Electronic portable devices such as mobile phones, USB flash discs, mp3 players, etc., which are worn close to the body, can represent personal dosemeters for members of the general public in situations of large-scale radiation accidents or malevolent acts with radioactive materials. This study investigated dosimetric properties of alumina substrates and aspects of using mobile phones as personal dosemeters. The alumina substrates exhibited favourable dosimetry characteristics. However, anomalous fading had to be properly corrected in order to achieve sufficient precision in dose estimate. Trial dose reconstruction performed by means of two mobile phones proved that mobile phones can be used for reconstruction of personal doses. (authors)

  12. Crystal Structure and Substrate Specificity of PTPN12

    Directory of Open Access Journals (Sweden)

    Hui Li

    2016-05-01

    Full Text Available PTPN12 is an important tumor suppressor that plays critical roles in various physiological processes. However, the molecular basis underlying the substrate specificity of PTPN12 remains uncertain. Here, enzymological and crystallographic studies have enabled us to identify two distinct structural features that are crucial determinants of PTPN12 substrate specificity: the pY+1 site binding pocket and specific basic charged residues along its surface loops. Key structurally plastic regions and specific residues in PTPN12 enabled recognition of different HER2 phosphorylation sites and regulated specific PTPN12 functions. In addition, the structure of PTPN12 revealed a CDK2 phosphorylation site in a specific PTPN12 loop. Taken together, our results not only provide the working mechanisms of PTPN12 for desphosphorylation of its substrates but will also help in designing specific inhibitors of PTPN12.

  13. Enzyme activity and kinetics in substrate-amended river sediments

    Energy Technology Data Exchange (ETDEWEB)

    Duddridge, J E; Wainwright, M

    1982-01-01

    In determining the effects of heavy metals in microbial activity and litter degradation in river sediments, one approach is to determine the effects of these pollutants on sediment enzyme activity and synthesis. Methods to assay amylase, cellulase and urease activity in diverse river sediments are reported. Enzyme activity was low in non-amended sediments, but increased markedly when the appropriate substrate was added, paralleling both athropogenic and natural amendment. Linear relationships between enzyme activity, length of incubation, sample size and substrate concentration were established. Sediment enzyme activity generally obeyed Michaelis-Menton kinetics, but of the three enzymes, urease gave least significant correlation coefficients when the data for substrate concentration versus activity was applied to the Eadie-Hofstee transformation of the Michaelis-Menten equation. K/sub m/ and V/sub max/ for amylase, cellulase and urease in sediments are reported. (JMT)

  14. Proposed cryogenic Q-factor measurement of mirror substrates

    Energy Technology Data Exchange (ETDEWEB)

    Nietzsche, Sandor; Zimmer, Anja; Vodel, Wolfgang; Thuerk, Matthias; Schmidl, Frank; Seidel, Paul [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, 07743 Jena (Germany)

    2004-03-07

    The thermal noise of optical components (e.g., end mirrors, beam splitters) is one of the limiting factors of the sensitivity of most of the present interferometric gravitational wave detectors, and it will be limiting in the advanced detectors now being designed. This thermal noise occurs mainly in the optical substrates and their mirror coatings. One possible method for minimizing thermal noise is cooling to cryogenic temperatures, maximizing the mechanical Q and maximizing the eigenfrequencies of the substrate. A new cryogenic apparatus for investigations of the temperature dependency of the Q-factor of several substrate materials down to 4.2 K is proposed. Possible methods of mode excitation and ring down measurement are discussed.

  15. On the use of Plexiglass Substrates for Neutron Mirrors

    International Nuclear Information System (INIS)

    Maayouf, R.M.A.

    2009-01-01

    The work deals with neutron reflectivity measurements performed for Ni films coated on different types of the commercially available plexiglass substrates. The Ni coatings were Ni Cr (80% Ni, 20% Cr), 58 Ni and natural nickel. The reflectivity behaviors of 58 Ni and natural nickel are compared with a present measurement performed for a 58 Ni film (150 nm thick) coated on glass substrate. Some of the present mirrors were measured several years before and are included in the presented measurements in order to check the quality of the plexiglass mirrors over years. It has been found, from the presented measurements, that plexiglass, as a substrate, successfully substitutes glass and the quality of the Ni coating can last for several years without deterioration.

  16. On mobility of ions in thin films in liquid substrates

    International Nuclear Information System (INIS)

    Matveev, Yu.A.

    1984-01-01

    The problem of energy dissipation by emission of ripplons is solved for an ion moving in the media with two interfaces (films on solid and liquid substrates), taking into account the Van der Waals interaction. It is shown that in contrast to the earlier considered case of solid substrate where the action of the Van der Waals forces causes only renormalization of the free fall acceleration, in the vase of liquid substrate the influence of these forces is much more complicated. In addition to renormalization of the amplitude of the emitted surface wave and change of the velocity threshold after which the wave mechanism is effective, in sufficiently thin film, where modes are ''intersected'' the analytical expressions for mobility are also significantly modified. In real experimental environments consideration of all the factors mentioned leads as a rule to higher ion mobility

  17. Mechanically flexible optically transparent porous mono-crystalline silicon substrate

    KAUST Repository

    Rojas, Jhonathan Prieto; Syed, Ahad A.; Hussain, Muhammad Mustafa

    2012-01-01

    For the first time, we present a simple process to fabricate a thin (≥5μm), mechanically flexible, optically transparent, porous mono-crystalline silicon substrate. Relying only on reactive ion etching steps, we are able to controllably peel off a thin layer of the original substrate. This scheme is cost favorable as it uses a low-cost silicon <100> wafer and furthermore it has the potential for recycling the remaining part of the wafer that otherwise would be lost and wasted during conventional back-grinding process. Due to its porosity, it shows see-through transparency and potential for flexible membrane applications, neural probing and such. Our process can offer flexible, transparent silicon from post high-thermal budget processed device wafer to retain the high performance electronics on flexible substrates. © 2012 IEEE.

  18. Low profile conformal antenna arrays on high impedance substrate

    CERN Document Server

    Singh, Hema; Jha, Rakesh Mohan

    2016-01-01

    This book presents electromagnetic (EM) design and analysis of dipole antenna array over high impedance substrate (HIS). HIS is a preferred substrate for low-profile antenna design, owing to its unique boundary conditions. Such substrates permit radiating elements to be printed on them without any disturbance in the radiation characteristics. Moreover HIS provides improved impedance matching, enhanced bandwidth, and increased broadside directivity owing to total reflection from the reactive surface and high input impedance. This book considers different configurations of HIS for array design on planar and non-planar high-impedance surfaces. Results are presented for cylindrical dipole, printed dipole, and folded dipole over single- and double-layered square-patch-based HIS and dogbone-based HIS. The performance of antenna arrays is analyzed in terms of performance parameters such as return loss and radiation pattern. The design presented shows acceptable return loss and mainlobe gain of radiation pattern. Thi...

  19. Optically transparent frequency selective surfaces on flexible thin plastic substrates

    Energy Technology Data Exchange (ETDEWEB)

    Dewani, Aliya A., E-mail: a.ashraf@griffith.edu.au; O’Keefe, Steven G.; Thiel, David V.; Galehdar, Amir [School Of Electrical Engineering, Griffith University, Brisbane, 4111 (Australia)

    2015-02-15

    A novel 2D simple low cost frequency selective surface was screen printed on thin (0.21 mm), flexible transparent plastic substrate (relative permittivity 3.2). It was designed, fabricated and tested in the frequency range 10-20 GHz. The plane wave transmission and reflection coefficients agreed with numerical modelling. The effective permittivity and thickness of the backing sheet has a significant effect on the frequency characteristics. The stop band frequency reduced from 15GHz (no backing) to 12.5GHz with polycarbonate. The plastic substrate thickness beyond 1.8mm has minimal effect on the resonant frequency. While the inner element spacing controls the stop-band frequency, the substrate thickness controls the bandwidth. The screen printing technique provided a simple, low cost FSS fabrication method to produce flexible, conformal, optically transparent and bio-degradable FSS structures which can find their use in electromagnetic shielding and filtering applications in radomes, reflector antennas, beam splitters and polarizers.

  20. Optically transparent frequency selective surfaces on flexible thin plastic substrates

    Science.gov (United States)

    Dewani, Aliya A.; O'Keefe, Steven G.; Thiel, David V.; Galehdar, Amir

    2015-02-01

    A novel 2D simple low cost frequency selective surface was screen printed on thin (0.21 mm), flexible transparent plastic substrate (relative permittivity 3.2). It was designed, fabricated and tested in the frequency range 10-20 GHz. The plane wave transmission and reflection coefficients agreed with numerical modelling. The effective permittivity and thickness of the backing sheet has a significant effect on the frequency characteristics. The stop band frequency reduced from 15GHz (no backing) to 12.5GHz with polycarbonate. The plastic substrate thickness beyond 1.8mm has minimal effect on the resonant frequency. While the inner element spacing controls the stop-band frequency, the substrate thickness controls the bandwidth. The screen printing technique provided a simple, low cost FSS fabrication method to produce flexible, conformal, optically transparent and bio-degradable FSS structures which can find their use in electromagnetic shielding and filtering applications in radomes, reflector antennas, beam splitters and polarizers.

  1. Substrate specificity within a family of outer membrane carboxylate channels.

    Directory of Open Access Journals (Sweden)

    Elif Eren

    2012-01-01

    Full Text Available Many Gram-negative bacteria, including human pathogens such as Pseudomonas aeruginosa, do not have large-channel porins. This results in an outer membrane (OM that is highly impermeable to small polar molecules, making the bacteria intrinsically resistant towards many antibiotics. In such microorganisms, the majority of small molecules are taken up by members of the OprD outer membrane protein family. Here we show that OprD channels require a carboxyl group in the substrate for efficient transport, and based on this we have renamed the family Occ, for outer membrane carboxylate channels. We further show that Occ channels can be divided into two subfamilies, based on their very different substrate specificities. Our results rationalize how certain bacteria can efficiently take up a variety of substrates under nutrient-poor conditions without compromising membrane permeability. In addition, they explain how channel inactivation in response to antibiotics can cause resistance but does not lead to decreased fitness.

  2. Uji produksi biosurfaktan oleh Pseudomonas sp. pada substrat yang berbeda

    Directory of Open Access Journals (Sweden)

    Fatimah Fatimah

    2012-02-01

    Full Text Available Biosurfactant, microbial metabolite whose properties like surfactant, was suggested to replace chemically synthesized surfactant for take in hand environtmental pollution by petroleum hydrocarbon. This work was done to examine potency of Pseudomonas sp. isolated from Tanjung Perak Harbor to produce biosurfactant. Also, to know the effect of different substrates (glucose + yeast extract, lubricating oil and hexadecane toward biosurfactant production. Pseudomonas sp. grown in mineral synthetic water and biosurfactant production was measured on stationary phase. Biosurfactant production based on emulsification activity and surface tension reduction of supernatant (using Du Nouy tensiometer. Solar, lubricating oil, and hexadecane were used to examine emulsification activity. Results indicated that Pseudomonas sp. have a potency to produce biosurfactant. Surface tension of supernatant decreased up to 20 dyne/cm, when grown on hexadecane substrate. Hexadecane is the best growing substrate for biosurfactant production than others.

  3. Transparent Substrates for Plasmonic Sensing by Lithography-Free Fabrication

    DEFF Research Database (Denmark)

    Thilsted, Anil Haraksingh

    This Ph.D. thesis presents fabrication and optimization of transparent plasmonic substrates that can be used for biological and chemical sensing by surface enhanced Raman spectroscopy (SERS) sensing and localized surface plasmon resonance refractive index (LSPR RI) sensing. These substrates are......-free fabrication methods, and resulted in large-area, high throughput and low cost production techniques. The fabrication techniques consisted of using aluminum patterned areas and reactive ion etching (RIE) to achieve nanopillars or nanocylinders in glass; using RIE to achieve nanopillars in silicon as a mould......, respectively. As the substrates were transparent, measurements from the backside were possible, showing a 44%, 1:7% and 71% Raman signal intensity in comparison to the measurements from the front, for the glass nanopillars, the polymer injected nanopillars and the transferred metal nanocaps, respectively...

  4. Highly reflective polymeric substrates functionalized utilizing atomic layer deposition

    Science.gov (United States)

    Zuzuarregui, Ana; Coto, Borja; Rodríguez, Jorge; Gregorczyk, Keith E.; Ruiz de Gopegui, Unai; Barriga, Javier; Knez, Mato

    2015-08-01

    Reflective surfaces are one of the key elements of solar plants to concentrate energy in the receivers of solar thermal electricity plants. Polymeric substrates are being considered as an alternative to the widely used glass mirrors due to their intrinsic and processing advantages, but optimizing both the reflectance and the physical stability of polymeric mirrors still poses technological difficulties. In this work, polymeric surfaces have been functionalized with ceramic thin-films by atomic layer deposition. The characterization and optimization of the parameters involved in the process resulted in surfaces with a reflection index of 97%, turning polymers into a real alternative to glass substrates. The solution we present here can be easily applied in further technological areas where seemingly incompatible combinations of polymeric substrates and ceramic coatings occur.

  5. Highly reflective polymeric substrates functionalized utilizing atomic layer deposition

    Energy Technology Data Exchange (ETDEWEB)

    Zuzuarregui, Ana, E-mail: a.zuzuarregui@nanogune.eu; Gregorczyk, Keith E. [CIC Nanogune Consolider, de Tolosa Hiribidea 76, 20018 San Sebastián (Spain); Coto, Borja; Ruiz de Gopegui, Unai; Barriga, Javier [IK4-Tekniker, Iñaki Goenaga 5, 20600 Eibar (Spain); Rodríguez, Jorge [Torresol Energy (SENER Group), Avda. de Zugazarte 61, 48930 Las Arenas (Spain); Knez, Mato [CIC Nanogune Consolider, de Tolosa Hiribidea 76, 20018 San Sebastián (Spain); IKERBASQUE Basque Foundation for Science, Maria Diaz de Haro 3, 48013 Bilbao (Spain)

    2015-08-10

    Reflective surfaces are one of the key elements of solar plants to concentrate energy in the receivers of solar thermal electricity plants. Polymeric substrates are being considered as an alternative to the widely used glass mirrors due to their intrinsic and processing advantages, but optimizing both the reflectance and the physical stability of polymeric mirrors still poses technological difficulties. In this work, polymeric surfaces have been functionalized with ceramic thin-films by atomic layer deposition. The characterization and optimization of the parameters involved in the process resulted in surfaces with a reflection index of 97%, turning polymers into a real alternative to glass substrates. The solution we present here can be easily applied in further technological areas where seemingly incompatible combinations of polymeric substrates and ceramic coatings occur.

  6. Highly reflective polymeric substrates functionalized utilizing atomic layer deposition

    International Nuclear Information System (INIS)

    Zuzuarregui, Ana; Gregorczyk, Keith E.; Coto, Borja; Ruiz de Gopegui, Unai; Barriga, Javier; Rodríguez, Jorge; Knez, Mato

    2015-01-01

    Reflective surfaces are one of the key elements of solar plants to concentrate energy in the receivers of solar thermal electricity plants. Polymeric substrates are being considered as an alternative to the widely used glass mirrors due to their intrinsic and processing advantages, but optimizing both the reflectance and the physical stability of polymeric mirrors still poses technological difficulties. In this work, polymeric surfaces have been functionalized with ceramic thin-films by atomic layer deposition. The characterization and optimization of the parameters involved in the process resulted in surfaces with a reflection index of 97%, turning polymers into a real alternative to glass substrates. The solution we present here can be easily applied in further technological areas where seemingly incompatible combinations of polymeric substrates and ceramic coatings occur

  7. Surprisingly high substrate specificities observed in complex biofilms

    DEFF Research Database (Denmark)

    Nierychlo, Marta; Kindaichi, Tomonori; Kragelund, Caroline

    The behavior of microorganisms in natural ecosystems (e.g. biofilms) differs significantly from laboratory studies. In nature microorganisms experience alternating periods of surplus nutrients, nutrient-limitation, and starvation. Literature data suggests that to survive and compete successfully......, microorganisms can regulate their metabolism expressing wide range of uptake and catabolic systems. However, ecophysiological studies of natural biofilms indicate that bacteria are very specialized in their choice of substrate, so even minor changes in substrate composition can affect the community composition...... by selection for different specialized species. We hypothesized that bacteria growing in natural environment express strongly conserved substrate specificity which is independent on short-term (few hours) variations in growth conditions. In this study, biofilm from Aalborg wastewater treatment plant was used...

  8. A pentacene monolayer trapped between graphene and a substrate.

    Science.gov (United States)

    Zhang, Qicheng; Peng, Boyu; Chan, Paddy Kwok Leung; Luo, Zhengtang

    2015-09-21

    A self-assembled pentacene monolayer can be fabricated between the solid-solid interface of few-layered graphene (FLG) and the mica substrate, through a diffusion-spreading method. By utilizing a transfer method that allows us to sandwich pentacene between graphene and mica, followed by controlled annealing, we enabled the diffused pentacene to be trapped in the interfaces and led to the formation of a stable monolayer. We found that the formation of a monolayer is kinetically favored by using a 2D Ising lattice gas model for pentacene trapped between the graphene-substrate interfaces. This kinetic Monte Carlo simulation results indicate that, due to the graphene substrate enclosure, the spreading of the first layer proceeds faster than the second layer, as the kinetics favors the filling of voids by molecules from the second layer. This graphene assisted monolayer assembly method provides a new avenue for the fabrication of two-dimensional monolayer structures.

  9. Recyclable organic solar cells on cellulose nanocrystal substrates.

    Science.gov (United States)

    Zhou, Yinhua; Fuentes-Hernandez, Canek; Khan, Talha M; Liu, Jen-Chieh; Hsu, James; Shim, Jae Won; Dindar, Amir; Youngblood, Jeffrey P; Moon, Robert J; Kippelen, Bernard

    2013-01-01

    Solar energy is potentially the largest source of renewable energy at our disposal, but significant advances are required to make photovoltaic technologies economically viable and, from a life-cycle perspective, environmentally friendly, and consequently scalable. Cellulose nanomaterials are emerging high-value nanoparticles extracted from plants that are abundant, renewable, and sustainable. Here, we report on the first demonstration of efficient polymer solar cells fabricated on optically transparent cellulose nanocrystal (CNC) substrates. The solar cells fabricated on the CNC substrates display good rectification in the dark and reach a power conversion efficiency of 2.7%. In addition, we demonstrate that these solar cells can be easily separated and recycled into their major components using low-energy processes at room temperature, opening the door for a truly recyclable solar cell technology. Efficient and easily recyclable organic solar cells on CNC substrates are expected to be an attractive technology for sustainable, scalable, and environmentally-friendly energy production.

  10. Composition of agarose substrate affects behavioral output of Drosophila larvae

    Directory of Open Access Journals (Sweden)

    Anthi Aristomenis Apostolopoulou

    2014-01-01

    Full Text Available In the last decade the Drosophila larva has evolved into a simple model organism offering the opportunity to integrate molecular genetics with systems neuroscience. This led to a detailed understanding of the functional neuronal networks for a number of sensory functions and behaviors including olfaction, vision, gustation and learning and memory. Typically, behavioral assays in use exploit simple Petri dish setups with either agarose or agar as a substrate. However, neither the quality nor the concentration of the substrate is generally standardized across these experiments and there is no data available on how larval behavior is affected by such different substrates. Here, we have investigated the effects of different agarose concentrations on several larval behaviors. We demonstrate that agarose concentration is an important parameter, which affects all behaviors tested: preference, feeding, learning and locomotion. Larvae can discriminate between different agarose concentrations, they feed differently on them, they can learn to associate an agarose concentration with an odor stimulus and crawl faster on a substrate of higher agarose concentration. Additionally, we have investigated the effect of agarose concentration on three quinine based behaviors: preference, feeding and learning. We show that in all cases examined the behavioral output changes in an agarose concentration-dependent manner. Our results suggest that comparisons between experiments performed on substrates differing in agarose concentration should be done with caution. It should be taken into consideration that the agarose concentration can affect the behavioral output and thereby the experimental outcomes per se potentially due to an increased escape response on more rigid substrates.

  11. Substrate matters: Magnetic tuning of the Fe monolayer

    Energy Technology Data Exchange (ETDEWEB)

    Ramanathan, A.A.; Khalifeh, J.M.

    2017-03-15

    The effect of substrate on the magnetism of the Fe monolayer (ML) is investigated using the total energy DFT calculations with the local spin density approximation (LSDA). The results show an in plane ferromagnetic coupling (FM) and a magnetic moment of 1.78 µ{sub B} for the relaxed Fe ML in the presence of the vanadium substrate. In comparison, the surface Fe(001) magnetic moment ranges between 2.97–3.01 µ{sub B}. This difference in the Fe surface moment of more than 1 µ{sub B} in the presence or absence of Vanadium allows tuning of the Fe magnetic moment and has great potential as a magnetic switch and in spintronic devices. The surface magnetic quenching of Fe with V is much more pronounced than with other transition metal substrates like Molybdenum or Tungsten. We have a reduction of 40.5% of the Fe (001) surface moment which is more than double the reduction obtained with the Fe/Mo(001) or the Fe/W(001) systems. The magnetic quenching is due to the strong hybridization between the Fe and V d bands. This is supported by the observed charge density redistribution and large inward relaxation of 18.37% for the Fe surface upon structural relaxation. The Fe ML is antiferromagnetically (AF) coupled with the V interfacial layer, which has an appreciable induced magnetic moment of 0.48 µ{sub B}. - Highlights: • We report the magnetic quenching of a Fe ML on V(001) substrate. • Almost double as compared to Mo and W substrates. • The Fe surface ML on V(001) shows FM ordering as opposed to AF ordering for Fe ML on Mo(001) and W(001) substrates.

  12. Influence of substrate and film thickness on polymer LIPSS formation

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Jing; Nogales, Aurora; Ezquerra, Tiberio A. [Instituto de Estructura de la Materia (IEM-CSIC), Serrano 121, Madrid 28006 (Spain); Rebollar, Esther, E-mail: e.rebollar@csic.es [Instituto de Química Física Rocasolano (IQFR-CSIC), Serrano 119, Madrid 28006 (Spain)

    2017-02-01

    Highlights: • The estimation of temperature upon pulse accumulation shows that a small positive offset is caused by each individual pulse. • Number of pulses needed for LIPSS formation in PS thin films depends on polymer thickness. • Thermal conductivity and diffusivity of supporting substrate influence the onset for LIPSS formation and their quality. • Quality of LIPSS is affected by the substrate optical properties. - Abstract: Here we focus on the influence of both, substrate and film thickness on polymer Laser Induced Periodic Surface Structures (LIPSS) formation in polymer films. For this aim a morphological description of ripples structures generated on spin-coated polystyrene (PS) films by a linearly polarized laser beam with a wavelength of 266 nm is presented. The influence of different parameters on the quality and characteristics of the formed laser-induced periodic surface structures (LIPSS) was investigated. We found that well-ordered LIPSS are formed either on PS films thinner than 200 nm or thicker than 400 nm supported on silicon substrates as well as on thicker free standing films. However less-ordered ripples are formed on silicon supported films with intermediate thicknesses in the range of 200–380 nm. The effect of the thermal and optical properties of the substrate on the quality of LIPSS was analyzed. Differences observed in the fluence and number of pulses needed for the onset of surface morphological modifications is explained considering two main effects which are: (1) The temperature increase on polymer surface induced by the action of cumulative laser irradiation and (2) The differences in thermal conductivity between the polymer and the substrate which strongly affect the heat dissipation generated by irradiation.

  13. Arabidopsis ATG4 cysteine proteases specificity toward ATG8 substrates

    Science.gov (United States)

    Park, Eunsook; Woo, Jongchan; Dinesh-Kumar, SP

    2014-01-01

    Macroautophagy (hereafter autophagy) is a regulated intracellular process during which cytoplasmic cargo engulfed by double-membrane autophagosomes is delivered to the vacuole or lysosome for degradation and recycling. Atg8 that is conjugated to phosphatidylethanolamine (PE) during autophagy plays an important role not only in autophagosome biogenesis but also in cargo recruitment. Conjugation of PE to Atg8 requires processing of the C-terminal conserved glycine residue in Atg8 by the Atg4 cysteine protease. The Arabidopsis plant genome contains 9 Atg8 (AtATG8a to AtATG8i) and 2 Atg4 (AtATG4a and AtATG4b) family members. To understand AtATG4’s specificity toward different AtATG8 substrates, we generated a unique synthetic substrate C-AtATG8-ShR (citrine-AtATG8-Renilla luciferase SuperhRLUC). In vitro analyses indicated that AtATG4a is catalytically more active and has broad AtATG8 substrate specificity compared with AtATG4b. Arabidopsis transgenic plants expressing the synthetic substrate C-AtAtg8a-ShR is efficiently processed by endogenous AtATG4s and targeted to the vacuole during nitrogen starvation. These results indicate that the synthetic substrate mimics endogenous AtATG8, and its processing can be monitored in vivo by a bioluminescence resonance energy transfer (BRET) assay. The synthetic Atg8 substrates provide an easy and versatile method to study plant autophagy during different biological processes. PMID:24658121

  14. Interface between Sn-Sb-Cu solder and copper substrate

    Energy Technology Data Exchange (ETDEWEB)

    Sebo, P., E-mail: Pavel.Sebo@savba.sk [Institute of Materials and Machine Mechanics, Slovak Academy of Sciences, Racianska 75, 831 02 Bratislava 3 (Slovakia); Svec, P. [Institute of Physics, Slovak Academy of Sciences, Dubravska cesta 9, 845 11 Bratislava 45 (Slovakia); Faculty of Materials Science and Technology, Slovak University of Technology, J. Bottu 25, 917 24 Trnava (Slovakia); Janickovic, D.; Illekova, E. [Institute of Physics, Slovak Academy of Sciences, Dubravska cesta 9, 845 11 Bratislava 45 (Slovakia); Plevachuk, Yu. [Ivan Franko National University, Department of Metal Physics, 79005 Lviv (Ukraine)

    2011-07-15

    Highlights: {yields} New lead-free solder materials based on Sn-Sb-Cu were designed and prepared. {yields} Melting and solidification temperatures of the solders have been determined. {yields} Cu-substrate/solder interaction has been analyzed and quantified. {yields} Phases formed at the solder-substrate interface have been identified. {yields} Composition and soldering atmospheres were correlated with joint strength. - Abstract: Influence of antimony and copper in Sn-Sb-Cu solder on the melting and solidification temperatures and on the microstructure of the interface between the solder and copper substrate after wetting the substrate at 623 K for 1800 s were studied. Microstructure of the interface between the solder and copper substrates in Cu-solder-Cu joints prepared at the same temperature for 1800 s was observed and shear strength of the joints was measured. Influence of the atmosphere - air with the flux and deoxidising N{sub 2} + 10H{sub 2} gas - was taken into account. Thermal stability and microstructure were studied by differential scanning calorimetry (DSC), light microscopy, scanning electron microscopy (SEM) with energy-dispersive spectrometry (EDS) and X-ray diffraction (XRD). Melting and solidification temperatures of the solders were determined. An interfacial transition zone was formed by diffusion reaction between solid copper and liquid solder. At the interface Cu{sub 3}Sn and Cu{sub 6}Sn{sub 5} phases arise. Cu{sub 3}Sn is adjacent to the Cu substrate and its thickness decreases with increasing the amount of copper in solder. Scallop Cu{sub 6}Sn{sub 5} phase is formed also inside the solder drop. The solid solution Sn(Sb) and SbSn phase compose the interior of the solder drop. Shear strength of the joints measured by push-off method decreases with increasing Sb concentration. Copper in the solder shows even bigger negative effect on the strength.

  15. Superhydrophobic SERS substrates based on silicon hierarchical nanostructures

    Science.gov (United States)

    Chen, Xuexian; Wen, Jinxiu; Zhou, Jianhua; Zheng, Zebo; An, Di; Wang, Hao; Xie, Weiguang; Zhan, Runze; Xu, Ningsheng; Chen, Jun; She, Juncong; Chen, Huanjun; Deng, Shaozhi

    2018-02-01

    Silicon nanostructures have been cultivated as promising surface enhanced Raman scattering (SERS) substrates in terms of their low-loss optical resonance modes, facile functionalization, and compatibility with today’s state-of-the-art CMOS techniques. However, unlike their plasmonic counterparts, the electromagnetic field enhancements induced by silicon nanostructures are relatively small, which restrict their SERS sensing limit to around 10-7 M. To tackle this problem, we propose here a strategy for improving the SERS performance of silicon nanostructures by constructing silicon hierarchical nanostructures with a superhydrophobic surface. The hierarchical nanostructures are binary structures consisted of silicon nanowires (NWs) grown on micropyramids (MPs). After being modified with perfluorooctyltriethoxysilane (PFOT), the nanostructure surface shows a stable superhydrophobicity with a high contact angle of ˜160°. The substrate can allow for concentrating diluted analyte solutions into a specific area during the evaporation of the liquid droplet, whereby the analytes are aggregated into a small volume and can be easily detected by the silicon nanostructure SERS substrate. The analyte molecules (methylene blue: MB) enriched from an aqueous solution lower than 10-8 M can be readily detected. Such a detection limit is ˜100-fold lower than the conventional SERS substrates made of silicon nanostructures. Additionally, the detection limit can be further improved by functionalizing gold nanoparticles onto silicon hierarchical nanostructures, whereby the superhydrophobic characteristics and plasmonic field enhancements can be combined synergistically to give a detection limit down to ˜10-11 M. A gold nanoparticle-functionalized superhydrophobic substrate was employed to detect the spiked melamine in liquid milk. The results showed that the detection limit can be as low as 10-5 M, highlighting the potential of the proposed superhydrophobic SERS substrate in

  16. Seneca Valley Virus 3Cpro Substrate Optimization Yields Efficient Substrates for Use in Peptide-Prodrug Therapy.

    Directory of Open Access Journals (Sweden)

    Linde A Miles

    Full Text Available The oncolytic picornavirus Seneca Valley Virus (SVV-001 demonstrates anti-tumor activity in models of small cell lung cancer (SCLC, but may ultimately need to be combined with cytotoxic therapies to improve responses observed in patients. Combining SVV-001 virotherapy with a peptide prodrug activated by the viral protease 3Cpro is a novel strategy that may increase the therapeutic potential of SVV-001. Using recombinant SVV-001 3Cpro, we measured cleavage kinetics of predicted SVV-001 3Cpro substrates. An efficient substrate, L/VP4 (kcat/KM = 1932 ± 183 M(-1s(-1, was further optimized by a P2' N→P substitution yielding L/VP4.1 (kcat/KM = 17446 ± 2203 M(-1s(-1. We also determined essential substrate amino acids by sequential N-terminal deletion and substitution of amino acids found in other picornavirus genera. A peptide corresponding to the L/VP4.1 substrate was selectively cleaved by SVV-001 3Cpro in vitro and was stable in human plasma. These data define an optimized peptide substrate for SVV-001 3Cpro, with direct implications for anti-cancer therapeutic development.

  17. System and process for aluminization of metal-containing substrates

    Energy Technology Data Exchange (ETDEWEB)

    Chou, Yeong-Shyung; Stevenson, Jeffry W.

    2017-12-12

    A system and method are detailed for aluminizing surfaces of metallic substrates, parts, and components with a protective alumina layer in-situ. Aluminum (Al) foil sandwiched between the metallic components and a refractory material when heated in an oxidizing gas under a compression load at a selected temperature forms the protective alumina coating on the surface of the metallic components. The alumina coating minimizes evaporation of volatile metals from the metallic substrates, parts, and components in assembled devices that can degrade performance during operation at high temperature.

  18. Substrates coated with silver nanoparticles as a neuronal regenerative material

    Directory of Open Access Journals (Sweden)

    Alon N

    2014-05-01

    Full Text Available Noa Alon,1,3,* Yana Miroshnikov,2,3,* Nina Perkas,2,3 Ifat Nissan,2,3 Aharon Gedanken,2,3 Orit Shefi1,31Faculty of Engineering, 2Department of Chemistry, 3Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat Gan, Israel*These authors contributed equally to this workAbstract: Much effort has been devoted to the design of effective biomaterials for nerve regeneration. Here, we report the novel use of silver nanoparticles (AgNPs as regenerative agents to promote neuronal growth. We grew neuroblastoma cells on surfaces coated with AgNPs and studied the effect on the development of the neurites during the initiation and the elongation growth phases. We find that the AgNPs function as favorable anchoring sites, and the growth on the AgNP-coated substrates leads to a significantly enhanced neurite outgrowth. Cells grown on substrates coated with AgNPs have initiated three times more neurites than cells grown on uncoated substrates, and two times more than cells grown on substrates sputtered with a plain homogenous layer of silver. The growth of neurites on AgNPs in the elongation phase was enhanced as well. A comparison with substrates coated with gold nanoparticles (AuNPs and zinc oxide nanoparticles (ZnONPs demonstrated a clear silver material-driven promoting effect, in addition to the nanotopography. The growth on substrates coated with AgNPs has led to a significantly higher number of initiating neurites when compared to substrates coated with AuNPs or ZnONPs. All nanoparticle-coated substrates affected and promoted the elongation of neurites, with a significant positive maximal effect for the AgNPs. Our results, combined with the well-known antibacterial effect of AgNPs, suggest the use of AgNPs as an attractive nanomaterial – with dual activity – for neuronal repair studies.Keywords: nerve regeneration, nanotopography, antibacterial material, neuroblastoma, gold nanoparticles, zinc oxide nanoparticles

  19. Fabrication of SERS Substrate by Multilayered Nanosphere Deposition Technique

    International Nuclear Information System (INIS)

    Fu, Chit Yaw; Dinish, U. S.; Praveen, Thoniyot; Koh, Zhen Yu; Kho, Khiang Wei; Malini, Olivo

    2010-01-01

    Metal film over nanosphere (MFON) has been employed as a reproducible and predictable SERS-active device in biosensing applications. In addition to its economic fabrication process, such substrate can be further processed to a prism-structure with increased SERS enhancement and wider Plasmon tunability. In this work, we investigate an alternative coating method to deposit a larger area of well-ordered PS beads with different sizes (oe = 100nm and 400 nm) onto a glass. The result suggests that the proposed well-coating technique can be suitably used to form closely-packed PS beads with diameter less than 100 nm for developing MFON substrates.

  20. Nickel W14 substrates for high-temperature superconductors

    International Nuclear Information System (INIS)

    Kolb-Telieps, Angelika; Gehrmann, Bodo

    2008-01-01

    High-temperature superconductivity is on the threshold of market launch. Starting from the results of a research project, ThyssenKrupp VDM has successfully developed an industrial-scale production process for nickel W14, which is used as substrate strip in superconductors destined for applications such as generators for wind turbines. The deoxidation of the melt presented a particular challenge. On the one hand, this is required in order to avoid fractures during the hot forming, on the other, the usual deoxidation elements have negative effects on the nanoscale texture and surface roughness needed for the substrate strip. (orig.)

  1. System and process for aluminization of metal-containing substrates

    Science.gov (United States)

    Chou, Yeong-Shyung; Stevenson, Jeffry W

    2015-11-03

    A system and method are detailed for aluminizing surfaces of metallic substrates, parts, and components with a protective alumina layer in-situ. Aluminum (Al) foil sandwiched between the metallic components and a refractory material when heated in an oxidizing gas under a compression load at a selected temperature forms the protective alumina coating on the surface of the metallic components. The alumina coating minimizes evaporation of volatile metals from the metallic substrates, parts, and components in assembled devices during operation at high temperature that can degrade performance.

  2. Mechanics of patterned helical Si springs on Si substrate.

    Science.gov (United States)

    Liu, D L; Ye, D X; Khan, F; Tang, F; Lim, B K; Picu, R C; Wang, G C; Lu, T M

    2003-12-01

    The elastic response, including the spring constant, of individual Si helical-shape submicron springs, was measured using a tip-cantilever assembly attached to a conventional atomic force microscope. The isolated, four-turn Si springs were fabricated using oblique angle deposition with substrate rotation, also known as the glancing angle deposition, on a templated Si substrate. The response of the structures was modeled using finite elements, and it was shown that the conventional formulae for the spring constant required modifications before they could be used for the loading scheme used in the present experiment.

  3. Ductile film delamination from compliant substrates using hard overlayers.

    Science.gov (United States)

    Cordill, M J; Marx, V M; Kirchlechner, C

    2014-11-28

    Flexible electronic devices call for copper and gold metal films to adhere well to polymer substrates. Measuring the interfacial adhesion of these material systems is often challenging, requiring the formulation of different techniques and models. Presented here is a strategy to induce well defined areas of delamination to measure the adhesion of copper films on polyimide substrates. The technique utilizes a stressed overlayer and tensile straining to cause buckle formation. The described method allows one to examine the effects of thin adhesion layers used to improve the adhesion of flexible systems.

  4. Influence of nutrition and various substrates on spruce seedling growth

    Directory of Open Access Journals (Sweden)

    Đukić Matilda

    2004-01-01

    Full Text Available The results of the influence of main macronutrients (N, P, and K on growth and development of spruce (Picea abies L. Karst one-year old seedlings are presented. They were grown in containers, in nursery conditions, on four different substrates. There is a good influence on biogenous element contents, height, root collar diameter, needle length and mass, root mass as well as physiological vitality of spruce seedlings. It was observed that the effect of nutrition depends also on the type of substrate.

  5. Optimization of Substrate Feeding for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias

    2013-01-01

    to be effective in mitigating the effects of substrate inhibition. Using enzymatic biodiesel production as a case study, the volumetric productivity of the reactor is increased while minimizing inactivation of the enzyme due to the alcohol. This is done by using a simple optimization routine where the substrate...... (both the vegetable oil and alcohol) feed rate/concentration is manipulated simultaneously. The results of the simulation were tested in the laboratory and are sufficiently positive to suggest the implementation of a feeding strategy for large scale enzymatic biodiesel production...

  6. Conductive and robust nitride buffer layers on biaxially textured substrates

    Science.gov (United States)

    Sankar, Sambasivan [Chicago, IL; Goyal, Amit [Knoxville, TN; Barnett, Scott A [Evanston, IL; Kim, Ilwon [Skokie, IL; Kroeger, Donald M [Knoxville, TN

    2009-03-31

    The present invention relates to epitaxial, electrically conducting and mechanically robust, cubic nitride buffer layers deposited epitaxially on biaxially textured substrates such as metals and alloys. The invention comprises of a biaxially textured substrate with epitaxial layers of nitrides. The invention also discloses a method to form such epitaxial layers using a high rate deposition method as well as without the use of forming gases. The invention further comprises epitaxial layers of oxides on the biaxially textured nitride layer. In some embodiments the article further comprises electromagnetic devices which may have superconducting properties.

  7. RFID and Memory Devices Fabricated Integrally on Substrates

    Science.gov (United States)

    Schramm, Harry F.

    2004-01-01

    Electronic identification devices containing radio-frequency identification (RFID) circuits and antennas would be fabricated integrally with the objects to be identified, according to a proposal. That is to say, the objects to be identified would serve as substrates for the deposition and patterning of the materials of the devices used to identify them, and each identification device would be bonded to the identified object at the molecular level. Vacuum arc vapor deposition (VAVD) is the NASA derived process for depositing layers of material on the substrate. This proposal stands in contrast to the current practice of fabricating RFID and/or memory devices as wafer-based, self-contained integrated-circuit chips that are subsequently embedded in or attached to plastic cards to make smart account-information cards and identification badges. If one relies on such a chip to store data on the history of an object to be tracked and the chip falls off or out of the object, then one loses both the historical data and the means to track the object and verify its identity electronically. Also, in contrast is the manufacturing philosophy in use today to make many memory devices. Today s methods involve many subtractive processes such as etching. This proposal only uses additive methods, building RFID and memory devices from the substrate up in thin layers. VAVD is capable of spraying silicon, copper, and other materials commonly used in electronic devices. The VAVD process sprays most metals and some ceramics. The material being sprayed has a very strong bond with the substrate, whether that substrate is metal, ceramic, or even wood, rock, glass, PVC, or paper. An object to be tagged with an identification device according to the proposal must be compatible with a vacuum deposition process. Temperature is seldom an issue as the substrate rarely reaches 150 F (66 C) during the deposition process. A portion of the surface of the object would be designated as a substrate for

  8. Laser assisted removal of fixed contamination from metallic substrate

    International Nuclear Information System (INIS)

    Kumar, Aniruddha; Prasad, Manisha; Prakash, Tej; Shail, Shailini; Bhatt, R.B.; Behere, P.G.; Mohd Afzal; Kumar, Arun; Biswas, D.J.

    2015-01-01

    A single mode pulsed fiber laser was used to remove fixed contamination from stainless steel substrate by ablation. Samples were simulated by electro-deposition technique with 232 U as the test contaminant. Laser power, repetition rate, laser beam scanning speed and number of passes were optimised to obtain the desired ablation depth in the substrate. Ablation depth varying between few microns to few hundreds of microns could be achieved through careful control of these processing parameters. The absence of any activity in laser treated samples provided experimental signature of the efficacy of the laser assisted removal of fixed contamination. (author)

  9. Recycled hydroxyapatite coatings on 316L stainless steel substrates

    International Nuclear Information System (INIS)

    Mendes Filho, Antonio Alves; Pereira, Renato Alves; Araujo, Fernando Gabriel da Silva; Sousa, Camila Mateus de

    2010-01-01

    In this work were evaluated recycled hydroxyapatite coatings on 316L stainless steel substrates by plasma thermal aspersion. The hydroxyapatite used was obtained from bovine bone by the hydrothermal method. The samples of hydroxyapatite powders were divided according to their particle size distribution. The adhesion of the powders coating to the substrate was evaluated by assay scratch. The X-ray diffraction techniques and scanning electron microscopy were also used. The results of scratch resistance were between 46N and 63N. Analysis by scanning electron microscopy and x-ray diffraction showed no cracks coatings, single-phase and with few fused particles. (author)

  10. A Simple Combinatorial Codon Mutagenesis Method for Targeted Protein Engineering.

    Science.gov (United States)

    Belsare, Ketaki D; Andorfer, Mary C; Cardenas, Frida S; Chael, Julia R; Park, Hyun June; Lewis, Jared C

    2017-03-17

    Directed evolution is a powerful tool for optimizing enzymes, and mutagenesis methods that improve enzyme library quality can significantly expedite the evolution process. Here, we report a simple method for targeted combinatorial codon mutagenesis (CCM). To demonstrate the utility of this method for protein engineering, CCM libraries were constructed for cytochrome P450 BM3 , pfu prolyl oligopeptidase, and the flavin-dependent halogenase RebH; 10-26 sites were targeted for codon mutagenesis in each of these enzymes, and libraries with a tunable average of 1-7 codon mutations per gene were generated. Each of these libraries provided improved enzymes for their respective transformations, which highlights the generality, simplicity, and tunability of CCM for targeted protein engineering.

  11. Crystallization and preliminary crystallographic analysis of porcine acylaminoacyl peptidase.

    Science.gov (United States)

    Wright, Helena; Kiss, András L; Szeltner, Zoltán; Polgár, László; Fülöp, Vilmos

    2005-10-01

    Acylaminoacyl peptidase (also known as acylamino-acid-releasing enzyme or acylpeptide hydrolase; EC 3.4.19.1) is an unusual member of the prolyl oligopeptidase family catalysing the hydrolysis of an N-acylated peptide to an acylamino acid and a peptide with a free N-terminus. Acylaminoacyl peptidase purified from porcine liver has been crystallized in mother liquor containing 0.1 M Tris-HCl pH 7.0, 10%(w/v) polyethylene glycol 8000, 50 mM MgCl2 and 1%(w/v) CHAPS using the hanging-drop vapour-diffusion technique. A full data set to 3.4 A resolution was collected at ESRF beamline ID14-4 and space group C222 was assigned, with unit-cell parameters a = 84.8, b = 421.1, c = 212.0 A and four molecules in the asymmetric unit.

  12. Low Defect Density Substrates and High-Quality Epi-Substrate Interfaces for ABCS Devices and Progress Toward Phonon-Mediated THz Lasers

    National Research Council Canada - National Science Library

    Goodhue, William; Bliss, David; Krishnaswami, Kannan; Vangala, Shivashankar; Li, Jin; Zhu, Beihong

    2005-01-01

    ... has been developing technology for producing low defect density substrates and high-quality epi-substrate interfaces for ABCS device applications as well as developing fabrication and device concepts...

  13. Effect of Feed Substrate Thickness on the Bandwidth and Radiation Characteristics of an Aperture-Coupled Microstrip Antenna with a High Permittivity Feed Substrate

    Directory of Open Access Journals (Sweden)

    Jae-Hyun Kim

    2018-04-01

    Full Text Available The impedance bandwidth and radiation characteristics of an aperture-coupled microstrip line-fed patch antenna (ACMPA with a high permittivity (ɛr = 10 feed substrate suitable for integration with a monolithic microwave integrated circuit (MMIC are investigated for various feed substrate thicknesses through an experiment and computer simulation. The impedance bandwidth of an ACMPA with a high permittivity feed substrate increases as the feed substrate thickness decreases. Furthermore, the front-to-back ratio of an ACMPA with a high permittivity feed substrate increases and the cross-polarization level decreases as the feed substrate thickness decreases. As the impedance bandwidth of an ACMPA with a high permittivity feed substrate increases and its radiation characteristics improve as the feed substrate thickness decreases, the ACMPA configuration becomes suitable for integration with an MMIC.

  14. Sexual dimorphism in the attachment ability of the ladybird beetle Coccinella septempunctata on soft substrates

    DEFF Research Database (Denmark)

    Heepe, Lars; Petersen, Dennis S.; Tölle, Lisa

    2017-01-01

    of insects and also that of spiders and geckos has been tested on rigid substrates only. However, the natural habitats of climbing animals may provide a variety of substrate stiffness ranging from rigid rock surfaces to soft, biofilm covered substrates. In order to test the effect of different substrate...

  15. Method of producing an electronic unit having a polydimethylsiloxane substrate and circuit lines

    Energy Technology Data Exchange (ETDEWEB)

    Davidson, James Courtney [Livermore, CA; Krulevitch, Peter A [Pleasanton, CA; Maghribi, Mariam N [Livermore, CA; Benett, William J [Livermore, CA; Hamilton, Julie K [Tracy, CA; Tovar, Armando R [San Antonio, TX

    2012-06-19

    A system of metalization in an integrated polymer microsystem. A flexible polymer substrate is provided and conductive ink is applied to the substrate. In one embodiment the flexible polymer substrate is silicone. In another embodiment the flexible polymer substrate comprises poly(dimethylsiloxane).

  16. Calcium-Dependent Protein Kinases from Arabidopsis show substrate specificity differences in an analysis of 103 substrates

    Directory of Open Access Journals (Sweden)

    Amy eCurran

    2011-08-01

    Full Text Available The identification of substrates represents a critical challenge for understanding any protein kinase-based signal transduction pathway. In Arabidopsis, there are more than 1000 different protein kinases, 34 of which belong to a family of Ca2+-dependent protein kinases (CPKs. While CPKs are implicated in regulating diverse aspects of plant biology, from ion transport to transcription, relatively little is known about isoform-specific differences in substrate specificity, or the number of phosphorylation targets. Here, in vitro kinase assays were used to compare phosphorylation targets of four CPKs from Arabidopsis (CPK1, 10, 16 and 34. Significant differences in substrate specificity for each kinase were revealed by assays using 103 different substrates. For example CPK16 phosphorylated Serine 109 in a peptide from the stress-regulated protein, Di19-2 with KM ~70 µM, but this site was not phosphorylated significantly by CPKs 1, 10, or 34. In contrast, CPKs 1, 10, and 34 phosphorylated 93 other peptide substrates not recognized by CPK16. Examples of substrate specificity differences among all four CPKs were verified by kinetic analyses. To test the correlation between in vivo phosphorylation events and in vitro kinase activities, assays were performed with 274 synthetic peptides that contained phosphorylation sites previously mapped in proteins isolated from plants (in vivo-mapped sites. Of these, 74 (27% were found to be phosphorylated by at least one of the four CPKs tested. This 27% success rate validates a robust strategy for linking the activities of specific kinases, such as CPKs, to the thousands of in planta phosphorylation sites that are being uncovered by emerging technologies.

  17. Hydrogenated amorphous silicon solar cells fabricated at low substrate temperature 110°C on flexible PET substrate

    Science.gov (United States)

    Ramakrishna, M.; Kumari, Juhi; Venkanna, K.; Agarwal, Pratima

    2018-05-01

    In this paper, we report a-Si:H solar cells fabricated on flexible Polyethylene terephthalate (PET) and corning glass. The a-Si:H thin films were prepared at low substrate temperature (110oC) on corning 1737 glass with different rf powers. The influence of rf power on structural and optoelectronic properties of i-a-Si:H were studied. The films deposited at rf power 50W show less broadening of peak. This indicates these films are more ordered. With this optimized parameter for i-layer, solar cells fabricated on flexible PET substrate show best efficiency of 3.3% whereas on corning glass 3.82%.

  18. Electron beam irradiating process for rendering rough or topographically irregular surface substrates smooth; and coated substrates produced thereby

    International Nuclear Information System (INIS)

    Nablo, S.V.

    1979-01-01

    This disclosure involves a novel process for instantaneous electron-beam curing of very thin low viscosity, solventless coating upon rough, irregular or textured surfaces of a substrate such as paper or the like. Through rather critical timing and energy adjustment procedures, the coating firmly adheres to the surface before the coating can conform to the roughness or texture contour or substantially penetrate into the surface. By this method a solidified very smooth outer surface is provided for the substrate that is particularly used for metalization and other finished layerings. (author)

  19. Photoinduced absorption of Ag nanoparticles deposited on ITO substrate

    International Nuclear Information System (INIS)

    Ozga, K.; Oyama, M.; Szota, M.; Nabialek, M.; Kityk, I.V.; Slezak, A.; Umar, A.A.; Nouneh, K.

    2011-01-01

    Research highlights: → We study photoinduced absorption for two Ag NP deposited on the ITO. → The higher resistance eof the NP favors larger photoinduced changes. → Principal role is played by nanointerfaces. - Abstract: Substantial changes of absorption after illumination by 300 mW continuous wave green laser at 532 nm were observed. The effect of indium tin oxide (ITO) substrate was explored versus Ag nanoparticles (AgNPs) size, their regularity and surface plasmon resonance. The ITO substrate features play a crucial role for the formation of homogenous AgNPs. The attachments of AgNPs on ITO surface as well as their homogeneity are significantly changed under the influence of the laser treatment. We study the Ag NP deposited on the two different substrates which play a crucial role in the photoinduced absorption. The dependence of the photoinduced absorption versus the time of optical treatment is explained within a framework of the photopolarization of the particular trapping levels on the borders between the ITO substrate and the Ag NP.

  20. Photoinduced absorption of Ag nanoparticles deposited on ITO substrate

    Energy Technology Data Exchange (ETDEWEB)

    Ozga, K., E-mail: cate.ozga@wp.pl [Chair of Public Health, Czestochowa University of Technology, Al. Armii Krajowej 36B, 42-200 Czestochowa (Poland); Oyama, M. [Department of Material Chemisrty, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8520 (Japan); Szota, M. [Institute of Materials Science and Engineering, Technical University of Czestochowa, al. Armii Krajowej 19, 42-200 Czestochowa (Poland); Nabialek, M. [Institute of Physics, Czestochowa University of Technology, Al. Armii Krajowej 19, 42-200 Czestochowa (Poland); Kityk, I.V. [Electrical Engineering Department, Czestochowa University of Technology, Al. Armii Krajowej 17/19, 42-200 Czestochowa (Poland); Slezak, A. [Chair of Public Health, Czestochowa University of Technology, Al. Armii Krajowej 36B, 42-200 Czestochowa (Poland); Umar, A.A. [Institute of Micronegineering and Nanoelectronics Universiti Kebangsaan Malaysia 43600 UKM bangi, Selangor D.E. (Malaysia); Nouneh, K. [INANOTECH, Institute of Nanomaterials and Nanotechnology, MAScIR (Moroccan Advanced Science, Innovation and Research Foundation), ENSET, Av. Armee Royale, 10100, Rabat (Morocco)

    2011-06-15

    Research highlights: > We study photoinduced absorption for two Ag NP deposited on the ITO. > The higher resistance eof the NP favors larger photoinduced changes. > Principal role is played by nanointerfaces. - Abstract: Substantial changes of absorption after illumination by 300 mW continuous wave green laser at 532 nm were observed. The effect of indium tin oxide (ITO) substrate was explored versus Ag nanoparticles (AgNPs) size, their regularity and surface plasmon resonance. The ITO substrate features play a crucial role for the formation of homogenous AgNPs. The attachments of AgNPs on ITO surface as well as their homogeneity are significantly changed under the influence of the laser treatment. We study the Ag NP deposited on the two different substrates which play a crucial role in the photoinduced absorption. The dependence of the photoinduced absorption versus the time of optical treatment is explained within a framework of the photopolarization of the particular trapping levels on the borders between the ITO substrate and the Ag NP.