Targeting Pin1 by inhibitor API-1 regulates microRNA biogenesis and suppresses hepatocellular carcinoma development.

Science.gov (United States)

Pu, Wenchen; Li, Jiao; Zheng, Yuanyuan; Shen, Xianyan; Fan, Xin; Zhou, Jian-Kang; He, Juan; Deng, Yulan; Liu, Xuesha; Wang, Chun; Yang, Shengyong; Chen, Qiang; Liu, Lunxu; Zhang, Guolin; Wei, Yu-Quan; Peng, Yong

2018-01-30

Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide, but there are few effective treatments. Aberrant microRNA (miRNA) biogenesis is correlated with HCC development. We previously demonstrated that peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) participates in miRNA biogenesis and is a potential HCC treatment target. However, how Pin1 modulates miRNA biogenesis remains obscure. Here, we present in vivo evidence that Pin1 overexpression is directly linked to the development of HCC. Administration with the Pin1 inhibitor (API-1), a specific small molecule targeting Pin1 peptidyl-prolyl isomerase domain and inhibiting Pin1 cis-trans isomerizing activity, suppresses in vitro cell proliferation and migration of HCC cells. But API-1-induced Pin1 inhibition is insensitive to HCC cells with low Pin1 expression and/or low exportin-5 (XPO5) phosphorylation. Mechanistically, Pin1 recognizes and isomerizes the phosphorylated serine-proline motif of phosphorylated XPO5 and passivates phosphorylated XPO5. Pin1 inhibition by API-1 maintains the active conformation of phosphorylated XPO5 and restores XPO5-driven precursor miRNA nuclear-to-cytoplasm export, activating anticancer miRNA biogenesis and leading to both in vitro HCC suppression and HCC suppression in xenograft mice. Experimental evidence suggests that Pin1 inhibition by API-1 up-regulates miRNA biogenesis by retaining active XPO5 conformation and suppresses HCC development, revealing the mechanism of Pin1-mediated miRNA biogenesis and unequivocally supporting API-1 as a drug candidate for HCC therapy, especially for Pin1-overexpressing, extracellular signal-regulated kinase-activated HCC. (Hepatology 2018). © 2018 by the American Association for the Study of Liver Diseases.

The prolyl isomerase Pin1 acts synergistically with CDK2 to regulate the basal activity of estrogen receptor α in breast cancer.

Directory of Open Access Journals (Sweden)

Chiara Lucchetti

Full Text Available In hormone receptor-positive breast cancers, most tumors in the early stages of development depend on the activity of the estrogen receptor and its ligand, estradiol. Anti-estrogens, such as tamoxifen, have been used as the first line of therapy for over three decades due to the fact that they elicit cell cycle arrest. Unfortunately, after an initial period, most cells become resistant to hormonal therapy. Peptidylprolyl isomerase 1 (Pin1, a protein overexpressed in many tumor types including breast, has been demonstrated to modulate ERalpha activity and is involved in resistance to hormonal therapy. Here we show a new mechanism through which CDK2 drives an ERalpha-Pin1 interaction under hormone- and growth factor-free conditions. The PI3K/AKT pathway is necessary to activate CDK2, which phosphorylates ERalphaSer294, and mediates the binding between Pin1 and ERalpha. Site-directed mutagenesis demonstrated that ERalphaSer294 is essential for Pin1-ERalpha interaction and modulates ERalpha phosphorylation on Ser118 and Ser167, dimerization and activity. These results open up new drug treatment opportunities for breast cancer patients who are resistant to anti-estrogen therapy.

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

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

Reduced-Amide Inhibitor of Pin1 Binds in a Conformation Resembling a Twisted-Amide Transition State†

Science.gov (United States)

Xu, Guoyan G.; Zhang, Yan; Mercedes-Camacho, Ana Y.; Etzkorn, Felicia A.

2011-01-01

The mechanism of the cell cycle regulatory peptidyl prolyl isomerase (PPIase), Pin1, was investigated using reduced-amide inhibitors designed to mimic the twisted-amide transition state. Inhibitors, R–pSer–Ψ[CH2N]–Pro–2-(indol-3-yl)-ethylamine, 1 (R = fluorenylmethoxycarbonyl, Fmoc), and 2 (R = Ac), of Pin1 were synthesized and bioassayed. Inhibitor 1 had an IC50 value of 6.3 μM, which is 4.5-fold better inhibition for Pin1 than our comparable ground state analogue, a cis-amide alkene isostere containing inhibitor. The change of Fmoc to Ac in 2 improved aqueous solubility for structural determination, and resulted in an IC50 value of 12 μM. The X-ray structure of the complex of 2 bound to Pin1 was determined to 1.76 Å resolution. The structure revealed that the reduced amide adopted a conformation similar to the proposed twisted-amide transition state of Pin1, with a trans-pyrrolidine conformation of the prolyl ring. A similar conformation of substrate would be destabilized relative to the planar amide conformation. Three additional reduced amides, with Thr replacing Ser, and l- or d-pipecolate (Pip) replacing Pro, were slightly weaker inhibitors of Pin1. PMID:21980916

A reduced-amide inhibitor of Pin1 binds in a conformation resembling a twisted-amide transition state.

Science.gov (United States)

Xu, Guoyan G; Zhang, Yan; Mercedes-Camacho, Ana Y; Etzkorn, Felicia A

2011-11-08

The mechanism of the cell cycle regulatory peptidyl prolyl isomerase (PPIase), Pin1, was investigated using reduced-amide inhibitors designed to mimic the twisted-amide transition state. Inhibitors, R-pSer-Ψ[CH(2)N]-Pro-2-(indol-3-yl)ethylamine, 1 [R = fluorenylmethoxycarbonyl (Fmoc)] and 2 (R = Ac), of Pin1 were synthesized and bioassayed. Inhibitor 1 had an IC(50) value of 6.3 μM, which is 4.5-fold better for Pin1 than our comparable ground-state analogue, a cis-amide alkene isostere-containing inhibitor. The change of Fmoc to Ac in 2 improved aqueous solubility for structural determination and resulted in an IC(50) value of 12 μM. The X-ray structure of the complex of 2 bound to Pin1 was determined to 1.76 Å resolution. The structure revealed that the reduced amide adopted a conformation similar to the proposed twisted-amide transition state of Pin1, with a trans-pyrrolidine conformation of the prolyl ring. A similar conformation of substrate would be destabilized relative to the planar amide conformation. Three additional reduced amides, with Thr replacing Ser and l- or d-pipecolate (Pip) replacing Pro, were slightly weaker inhibitors of Pin1.

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.

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

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.

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.

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

Complete determination of the Pin1 catalytic domain thermodynamic cycle by NMR lineshape analysis

International Nuclear Information System (INIS)

Greenwood, Alexander I.; Rogals, Monique J.; De, Soumya; Lu, Kun Ping; Kovrigin, Evgenii L.; Nicholson, Linda K.

2011-01-01

The phosphorylation-specific peptidyl-prolyl isomerase Pin1 catalyzes the isomerization of the peptide bond preceding a proline residue between cis and trans isomers. To best understand the mechanisms of Pin1 regulation, rigorous enzymatic assays of isomerization are required. However, most measures of isomerase activity require significant constraints on substrate sequence and only yield rate constants for the cis isomer, k cat cis and apparent Michaelis constants, K M App . By contrast, NMR lineshape analysis is a powerful tool for determining microscopic rates and populations of each state in a complex binding scheme. The isolated catalytic domain of Pin1 was employed as a first step towards elucidating the reaction scheme of the full-length enzyme. A 24-residue phosphopeptide derived from the amyloid precurser protein intracellular domain (AICD) phosphorylated at Thr668 served as a biologically-relevant Pin1 substrate. Specific 13 C labeling at the Pin1-targeted proline residue provided multiple reporters sensitive to individual isomer binding and on-enzyme catalysis. We have performed titration experiments and employed lineshape analysis of phosphopeptide 13 C– 1 H constant time HSQC spectra to determine k cat cis , k cat trans , K D cis , and K D trans for the catalytic domain of Pin1 acting on this AICD substrate. The on-enzyme equilibrium value of [E·trans]/[E·cis] = 3.9 suggests that the catalytic domain of Pin1 is optimized to operate on this substrate near equilibrium in the cellular context. This highlights the power of lineshape analysis for determining the microscopic parameters of enzyme catalysis, and demonstrates the feasibility of future studies of Pin1-PPIase mutants to gain insights on the catalytic mechanism of this important enzyme.

Extended Impact of Pin1 Catalytic Loop Phosphorylation Revealed by S71E Phosphomimetic.

Science.gov (United States)

Mahoney, Brendan J; Zhang, Meiling; Zintsmaster, John S; Peng, Jeffrey W

2018-03-02

Pin1 is a two-domain human protein that catalyzes the cis-trans isomerization of phospho-Ser/Thr-Pro (pS/T-P) motifs in numerous cell-cycle regulatory proteins. These pS/T-P motifs bind to Pin1's peptidyl-prolyl isomerase (PPIase) domain in a catalytic pocket, between an extended catalytic loop and the PPIase domain core. Previous studies showed that post-translational phosphorylation of S71 in the catalytic loop decreases substrate binding affinity and isomerase activity. To define the origins for these effects, we investigated a phosphomimetic Pin1 mutant, S71E-Pin1, using solution NMR. We find that S71E perturbs not only its host loop but also the nearby PPIase core. The perturbations identify a local network of hydrogen bonds and salt bridges that is more extended than previously thought, and includes interactions between the catalytic loop and the α2/α3 turn in the PPIase core. Explicit-solvent molecular dynamics simulations and phylogenetic analysis suggest that these interactions act as conserved "latches" between the loop and PPIase core that enhance binding of phosphorylated substrates, as they are absent in PPIases lacking pS/T-P specificity. Our results suggest that S71 is a hub residue within an electrostatic network primed for phosphorylation, and may illustrate a common mechanism of phosphorylation-mediated allostery. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

Role of Pin1 in UVA-induced cell proliferation and malignant transformation in epidermal cells

International Nuclear Information System (INIS)

Han, Chang Yeob; Hien, Tran Thi; Lim, Sung Chul; Kang, Keon Wook

2011-01-01

Highlights: → Pin1 expression is enhanced by low energy UVA irradiation in both skin tissues of hairless mice and JB6 C141 epidermal cells. → UVA irradiation increases activator protein-1 activity and cyclin D1 in a Pin1-dependent manner. → UVA potentiates EGF-inducible, anchorage-independent growth of epidermal cells, and this is suppressed by Pin1 inhibition or by anti-oxidant. -- Abstract: Ultraviolet A (UVA) radiation (λ = 320-400 nm) is considered a major cause of human skin cancer. Pin1, a peptidyl prolyl isomerase, is overexpressed in most types of cancer tissues and plays an important role in cell proliferation and transformation. Here, we demonstrated that Pin1 expression was enhanced by low energy UVA (300-900 mJ/cm 2 ) irradiation in both skin tissues of hairless mice and JB6 C141 epidermal cells. Exposure of epidermal cells to UVA radiation increased cell proliferation and cyclin D1 expression, and these changes were blocked by Pin1 inhibition. UVA irradiation also increased activator protein-1 (AP-1) minimal reporter activity and nuclear levels of c-Jun, but not c-Fos, in a Pin1-dependent manner. The increases in Pin1 expression and in AP-1 reporter activity in response to UVA were abolished by N-acetylcysteine (NAC) treatment. Finally, we found that pre-exposure of JB6 C141 cells to UVA potentiated EGF-inducible, anchorage-independent growth, and this effect was significantly suppressed by Pin1inhibition or by NAC.

DLX5, FGF8 and the Pin1 isomerase control ΔNp63α protein stability during limb development: a regulatory loop at the basis of the SHFM and EEC congenital malformations

Science.gov (United States)

Restelli, Michela; Lopardo, Teresa; Lo Iacono, Nadia; Garaffo, Giulia; Conte, Daniele; Rustighi, Alessandra; Napoli, Marco; Del Sal, Giannino; Perez-Morga, David; Costanzo, Antonio; Merlo, Giorgio Roberto; Guerrini, Luisa

2014-01-01

Ectrodactyly, or Split-Hand/Foot Malformation (SHFM), is a congenital condition characterized by the loss of central rays of hands and feet. The p63 and the DLX5;DLX6 transcription factors, expressed in the embryonic limb buds and ectoderm, are disease genes for these conditions. Mutations of p63 also cause the ectodermal dysplasia–ectrodactyly–cleft lip/palate (EEC) syndrome, comprising SHFM. Ectrodactyly is linked to defects of the apical ectodermal ridge (AER) of the developing limb buds. FGF8 is the key signaling molecule in this process, able to direct proximo-distal growth and patterning of the skeletal primordial of the limbs. In the limb buds of both p63 and Dlx5;Dlx6 murine models of SHFM, the AER is poorly stratified and FGF8 expression is severely reduced. We show here that the FGF8 locus is a downstream target of DLX5 and that FGF8 counteracts Pin1–ΔNp63α interaction. In vivo, lack of Pin1 leads to accumulation of the p63 protein in the embryonic limbs and ectoderm. We show also that ΔNp63α protein stability is negatively regulated by the interaction with the prolyl-isomerase Pin1, via proteasome-mediated degradation; p63 mutant proteins associated with SHFM or EEC syndromes are resistant to Pin1 action. Thus, DLX5, p63, Pin1 and FGF8 participate to the same time- and location-restricted regulatory loop essential for AER stratification, hence for normal patterning and skeletal morphogenesis of the limb buds. These results shed new light on the molecular mechanisms at the basis of the SHFM and EEC limb malformations. PMID:24569166

DLX5, FGF8 and the Pin1 isomerase control ΔNp63α protein stability during limb development: a regulatory loop at the basis of the SHFM and EEC congenital malformations.

Science.gov (United States)

Restelli, Michela; Lopardo, Teresa; Lo Iacono, Nadia; Garaffo, Giulia; Conte, Daniele; Rustighi, Alessandra; Napoli, Marco; Del Sal, Giannino; Perez-Morga, David; Costanzo, Antonio; Merlo, Giorgio Roberto; Guerrini, Luisa

2014-07-15

Ectrodactyly, or Split-Hand/Foot Malformation (SHFM), is a congenital condition characterized by the loss of central rays of hands and feet. The p63 and the DLX5;DLX6 transcription factors, expressed in the embryonic limb buds and ectoderm, are disease genes for these conditions. Mutations of p63 also cause the ectodermal dysplasia-ectrodactyly-cleft lip/palate (EEC) syndrome, comprising SHFM. Ectrodactyly is linked to defects of the apical ectodermal ridge (AER) of the developing limb buds. FGF8 is the key signaling molecule in this process, able to direct proximo-distal growth and patterning of the skeletal primordial of the limbs. In the limb buds of both p63 and Dlx5;Dlx6 murine models of SHFM, the AER is poorly stratified and FGF8 expression is severely reduced. We show here that the FGF8 locus is a downstream target of DLX5 and that FGF8 counteracts Pin1-ΔNp63α interaction. In vivo, lack of Pin1 leads to accumulation of the p63 protein in the embryonic limbs and ectoderm. We show also that ΔNp63α protein stability is negatively regulated by the interaction with the prolyl-isomerase Pin1, via proteasome-mediated degradation; p63 mutant proteins associated with SHFM or EEC syndromes are resistant to Pin1 action. Thus, DLX5, p63, Pin1 and FGF8 participate to the same time- and location-restricted regulatory loop essential for AER stratification, hence for normal patterning and skeletal morphogenesis of the limb buds. These results shed new light on the molecular mechanisms at the basis of the SHFM and EEC limb malformations. © The Author 2014. Published by Oxford University Press.

Pin1 and secondary hyperparathyroidism of chronic kidney disease: gene polymorphisms and protein levels.

Science.gov (United States)

Zhao, Yu; Zhang, Li-Li; Ding, Fa-Xian; Cao, Ping; Qi, Yuan-Yuan; Wang, Jing

2017-11-01

Peptidyl-prolyl cis/trans isomerase NIMA-interacting 1 (Pin1) is a key regulator of PTH mRNA stability. Secondary hyperparathyroidism (SHPT), which is characterized by elevated serum PTH levels, is a common complication of CKD. We investigated the possible associations between CKD with SHPT (CKD SHPT) and single-nucleotide polymorphisms of the Pin1 gene and compared the levels of the Pin1 protein in the CKD SHPT patients with those of the controls. The study group included 251 CKD SHPT patients and 61 controls. One putative functional SNP (single nucleotide polymorphism) in the Pin1 promoter (rs2233679C > T: c.-667C > T) is the main object. Genotyping was performed on purified DNA using polymerase chain reaction-restriction (PCR) and restriction fragment length polymorphisms (RFLP). The levels of Pin1 were measured in serum using an enzyme-linked immunosorbent assay. Genotyping showed that CT + TT in the Pin1 promoter was significantly more common in the CKD SHPT group than in the control group (p<.05). The correlation analysis demonstrated that a significant difference in the C to T transition in the Pin1 promoter contributed to CKD SHPT (χ 2 =12.47, p<.05; Odds ratios (OR) = 1.26, 95% confidence (CI) intervals =1.06-1.49). The multivariate logistic regression analysis reported that the OR and 95%CI were 12.693 and 2.029-75.819 (p<.05), respectively, in the Pin1 gene promoter -667T variant genotypes (CT + TT) after adjusting for other factors, and those values in Pin1 were 0.310 and 0.122-0.792 (p<.05). The -667T genetic variants in the Pin1 promoter contribute to an increased risk of CKD SHPT and may be biomarkers of susceptibility to CKD SHPT.

Neighboring phosphoSer-Pro motifs in the undefined domain of IRAK1 impart bivalent advantage for Pin1 binding.

Science.gov (United States)

Rogals, Monique J; Greenwood, Alexander I; Kwon, Jeahoo; Lu, Kun Ping; Nicholson, Linda K

2016-12-01

The peptidyl prolyl isomerase Pin1 has two domains that are considered to be its binding (WW) and catalytic (PPIase) domains, both of which interact with phosphorylated Ser/Thr-Pro motifs. This shared specificity might influence substrate selection, as many known Pin1 substrates have multiple sequentially close phosphoSer/Thr-Pro motifs, including the protein interleukin-1 receptor-associated kinase-1 (IRAK1). The IRAK1 undefined domain (UD) contains two sets of such neighboring motifs (Ser131/Ser144 and Ser163/Ser173), suggesting possible bivalent interactions with Pin1. Using a series of NMR titrations with 15N-labeled full-length Pin1 (Pin1-FL), PPIase, or WW domain and phosphopeptides representing the Ser131/Ser144 and Ser163/Ser173 regions of IRAK1-UD, bivalent interactions were investigated. Binding studies using singly phosphorylated peptides showed that individual motifs displayed weak affinities (> 100 μm) for Pin1-FL and each isolated domain. Analysis of dually phosphorylated peptides binding to Pin1-FL showed that inclusion of bivalent states was necessary to fit the data. The resulting complex model and fitted parameters were applied to predict the impact of bivalent states at low micromolar concentrations, demonstrating significant affinity enhancement for both dually phosphorylated peptides (3.5 and 24 μm for peptides based on the Ser131/Ser144 and Ser163/Ser173 regions, respectively). The complementary technique biolayer interferometry confirmed the predicted affinity enhancement for a representative set of singly and dually phosphorylated Ser131/Ser144 peptides at low micromolar concentrations, validating model predictions. These studies provide novel insights regarding the complexity of interactions between Pin1 and activated IRAK1, and more broadly suggest that phosphorylation of neighboring Ser/Thr-Pro motifs in proteins might provide competitive advantage at cellular concentrations for engaging with Pin1. © 2016 Federation of European

Discovery of novel selenium derivatives as Pin1 inhibitors by high-throughput screening

International Nuclear Information System (INIS)

Subedi, Amit; Shimizu, Takeshi; Ryo, Akihide; Sanada, Emiko; Watanabe, Nobumoto; Osada, Hiroyuki

2016-01-01

Peptidyl prolyl cis/trans isomerization by Pin1 regulates various oncogenic signals during cancer progression, and its inhibition through multiple approaches has established Pin1 as a therapeutic target. However, lack of simplified screening systems has limited the discovery of potent Pin1 inhibitors. We utilized phosphorylation-dependent binding of Pin1 to its specific substrate to develop a screening system for Pin1 inhibitors. Using this system, we screened a chemical library, and identified a novel selenium derivative as Pin1 inhibitor. Based on structure-activity guided chemical synthesis, we developed more potent Pin1 inhibitors that inhibited cancer cell proliferation. -- Highlights: •Novel screening for Pin1 inhibitors based on Pin1 binding is developed. •A novel selenium compound is discovered as Pin1 inhibitor. •Activity guided chemical synthesis of selenium derivatives resulted potent Pin1 inhibitors.

Pin1, a new player in the fate of HIF-1α degradation: an hypothetical mechanism inside vascular damage as Alzheimer’s disease risk factor.

Directory of Open Access Journals (Sweden)

Elena eLonati

2014-01-01

Full Text Available Aetiology of neurodegenerative mechanisms underlying Alzheimer's disease (AD are still under elucidation. The contribution of cerebrovascular deficiencies (such as cerebral ischemia/stroke has been strongly endorsed in recent years. Reduction of blood supply leading to hypoxic condition is known to activate cellular responses mainly controlled by hypoxia-inducible transcription factor-1 (HIF-1. Thus alterations of oxygen responsive HIF-1α subunit in the central nervous system may contribute to the cognitive decline, especially influencing mechanisms associated to APP (amyloid precursor protein amyloidogenic metabolism. Although HIF-1α protein level is known to be regulated by von Hippel-Lindau (VHL ubiquitin-proteasome system, it has been recently suggested that Gsk-3β (glycogen synthase kinase-3β promotes a VHL-independent HIF-1α degradation. Here we provide evidences that in rat primary hippocampal cell cultures, HIF-1α degradation might be mediated by a synergic action of Gsk-3β and Pin1 (peptidyl-prolyl cis/trans isomerase. In post-ischemic conditions, such as those mimicked with oxygen glucose deprivation (OGD, HIF-1α protein level increases remaining unexpectedly high for long time after normal condition restoration jointly with the increase of LDH (lactate dehydrogenase and BACE1 (β-secretase 1 protein expression (70% and 140% respectively. Interestingly the Pin1 activity decreases about 40%-60% and Pin1S16 inhibitory phosphorylation significantly increases, indicating that Pin1 binding to its substrate and enzymatic activity are reduced by treatment. Co-immunoprecipitation experiments demonstrate that HIF-1α/Pin1 in normoxia are associated, and that in presence of specific Pin1 and Gsk-3β inhibitors their interaction is reduced in parallel to an increase of HIF-1α protein level. Thus we suggest that in post-OGD neurons the high level of HIF-1α might be due to Pin1 binding ability and activity reduction which affects HIF-1�

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

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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

Calpastatin is regulated by protein never in mitosis gene A interacting-1 (PIN1) in endothelial cells

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Liu, Tongzheng, E-mail: liu.tongzheng@mayo.edu [Division of Oncology Research, Department of Oncology, Mayo Clinic, Rochester, MN 55905 (United States); Schneider, Ryan A., E-mail: schneiderr@findlay.edu [College of Pharmacy, The University of Findlay, Findlay, OH 45840 (United States); Hoyt, Dale G., E-mail: hoyt.27@osu.edu [The Dorothy M. Davis Heart and Lung Research Institute, and the Division of Pharmacology, College of Pharmacy, The Ohio State University, 500 West Twelfth Avenue, Columbus, OH 43210 (United States)

2011-10-28

Highlights: Black-Right-Pointing-Pointer Depletion of PIN1 increases inhibitory effect of calpastatin against calpain in endothelial cells. Black-Right-Pointing-Pointer PIN1 associates with calpastatin. Black-Right-Pointing-Pointer PIN1, but not mutants, reduces the inhibitory activity of calpastatin in vitro. Black-Right-Pointing-Pointer Depletion of calpastatin shows that it is required for PIN1 depletion to reduce calpain activity. -- Abstract: The peptidyl-proline isomerase, protein never in mitosis gene A interacting-1 (PIN1) binds and isomerizes proteins phosphorylated on serine/threonine before a proline. It was previously found that depletion of PIN1 greatly increased induction of cyclooxygenase-2 and inducible nitric oxide synthase by lowering calpain activity in murine aortic endothelial cells (MAEC). Here we investigated the effect of PIN1 on the endogenous inhibitor of heterodimeric {mu}- and m-calpains, calpastatin. MAEC were transduced with small hairpin (sh) RNA to knock down PIN1 (KD) or an inactive Control shRNA. Cells were also treated with non-targeted double stranded small inhibitory RNA (siRNA) or siRNA designed to deplete calpastatin. Despite reducing calpain activity, PIN1 KD did not significantly affect the expression of {mu}- and m-calpains, or calpastatin, compared to Control shRNA. Instead, depletion of PIN1 increased the inhibitory activity of calpastatin. Calpastatin co-immunoprecipitated with endogenous PIN1 and was pulled down with glutathione-S-transferase (GST)-PIN1 fusion protein. Adding GST-PIN1 to KD cell extracts lacking PIN1 reduced calpastatin inhibitory activity. Substrate binding and catalytic domain mutants of PIN1 failed to do so. These results suggest that protein interaction and the proline isomerase functions of PIN1 are required for it to inhibit calpastatin. Furthermore, depletion of calpastatin raised calpain activity and reduced calpain inhibitory activity to similar levels in KD and Control MAEC, indicating that

Calpastatin is regulated by protein never in mitosis gene A interacting-1 (PIN1) in endothelial cells

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Liu, Tongzheng; Schneider, Ryan A.; Hoyt, Dale G.

2011-01-01

Highlights: ► Depletion of PIN1 increases inhibitory effect of calpastatin against calpain in endothelial cells. ► PIN1 associates with calpastatin. ► PIN1, but not mutants, reduces the inhibitory activity of calpastatin in vitro. ► Depletion of calpastatin shows that it is required for PIN1 depletion to reduce calpain activity. -- Abstract: The peptidyl-proline isomerase, protein never in mitosis gene A interacting-1 (PIN1) binds and isomerizes proteins phosphorylated on serine/threonine before a proline. It was previously found that depletion of PIN1 greatly increased induction of cyclooxygenase-2 and inducible nitric oxide synthase by lowering calpain activity in murine aortic endothelial cells (MAEC). Here we investigated the effect of PIN1 on the endogenous inhibitor of heterodimeric μ- and m-calpains, calpastatin. MAEC were transduced with small hairpin (sh) RNA to knock down PIN1 (KD) or an inactive Control shRNA. Cells were also treated with non-targeted double stranded small inhibitory RNA (siRNA) or siRNA designed to deplete calpastatin. Despite reducing calpain activity, PIN1 KD did not significantly affect the expression of μ- and m-calpains, or calpastatin, compared to Control shRNA. Instead, depletion of PIN1 increased the inhibitory activity of calpastatin. Calpastatin co-immunoprecipitated with endogenous PIN1 and was pulled down with glutathione-S-transferase (GST)–PIN1 fusion protein. Adding GST–PIN1 to KD cell extracts lacking PIN1 reduced calpastatin inhibitory activity. Substrate binding and catalytic domain mutants of PIN1 failed to do so. These results suggest that protein interaction and the proline isomerase functions of PIN1 are required for it to inhibit calpastatin. Furthermore, depletion of calpastatin raised calpain activity and reduced calpain inhibitory activity to similar levels in KD and Control MAEC, indicating that calpastatin is required for PIN1 depletion to lower calpain activity. Thus, PIN1 apparently restrains

Loss of Pin1 Suppresses Hedgehog-Driven Medulloblastoma Tumorigenesis

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Tao Xu

2017-03-01

Full Text Available Medulloblastoma is the most common malignant brain tumor in children. Therapeutic approaches to medulloblastoma (combination of surgery, radiotherapy, and chemotherapy have led to significant improvements, but these are achieved at a high cost to quality of life. Alternative therapeutic approaches are needed. Genetic mutations leading to the activation of the Hedgehog pathway drive tumorigenesis in ~30% of medulloblastoma. In a yeast two-hybrid proteomic screen, we discovered a novel interaction between GLI1, a key transcription factor for the mediation of Hedgehog signals, and PIN1, a peptidylprolyl cis/trans isomerase that regulates the postphosphorylation fate of its targets. The GLI1/PIN1 interaction was validated by reciprocal pulldowns using epitope-tagged proteins in HEK293T cells as well as by co-immunoprecipiations of the endogenous proteins in a medulloblastoma cell line. Our results support a molecular model in which PIN1 promotes GLI1 protein abundance, thus contributing to the positive regulation of Hedgehog signals. Most importantly, in vivo functional analyses of Pin1 in the GFAP-tTA;TRE-SmoA1 mouse model of Hedgehog-driven medulloblastoma demonstrate that the loss of Pin1 impairs tumor development and dramatically increases survival. In summary, the discovery of the GLI1/PIN1 interaction uncovers PIN1 as a novel therapeutic target in Hedgehog-driven medulloblastoma tumorigenesis.

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.

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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.

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

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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

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

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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.

The polymorphism and haplotypes of PIN1 gene are associated with the risk of lung cancer in Southern and Eastern Chinese populations.

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Lu, Jiachun; Yang, Lei; Zhao, Hongjun; Liu, Bin; Li, Yinyan; Wu, Hongxia; Li, Qingchu; Zeng, Bohang; Wang, Yunnan; Ji, Weidong; Zhou, Yifeng

2011-11-01

Peptidyl-prolyl cis/trans isomerase (PPIase), PIN1, has been found to be a critical catalyst that involves in multiple oncogenic signaling pathways. Recently, several putative functional polymorphisms of the PIN1 gene have been identified to be associated with cancer risk. In this study, we tested the hypothesis that two common polymorphisms, c.-842G>C (rs2233678) and c.-667C>T (rs2233679), in the PIN1 promoter are associated with risk of lung cancer. In two independent case-control studies of 1,559 lung cancer cases and 1,679 controls conducted in Southern and Eastern Chinese population, we found that compared with the most common c.-842GG genotype, the carriers of c.-842C variant genotypes (GC + CC) had a decreased risk of lung cancer (odds ratio [OR] = 0.63, 95% confidence interval [CI] = 0.51-0.78, p = 1.13 × 10(-5) ). Although no association was observed between the c.-667C>T polymorphism and cancer risk, we found that the haplotype "C-C" had a greater protective effect (OR = 0.39, 95% CI = 0.23-0.67, p = 5.03 × 10(-4) ). The stratification analysis showed that the protective role of c.-842C variants was more pronounced in current smokers (p = 4.45 × 10(-5) ), especially in male smokers (p = 6.71 × 10(-6) ) and in those who smoked more than 20 pack-years (p = 2.30 × 10(-5) ) and the c.-842C variant genotypes interacted with smoking status (P(interaction) = 0.019) or pack-years smoked (P(interaction) = 0.008) on reducing cancer risk. Further functional assay revealed that the c.-842C variant allele had a lower transcription activity in luciferase assay and a lower DNA-binding ability with nuclear proteins, and low transcription activity in western blot assay. In conclusions, our data suggest that functional c.-842C variants and haplotype "C-C" in the PIN1 promoter contribute to decreased risk of lung cancer by diminishing the promoter activity, which may be susceptibility biomarkers for lung cancer. © 2011 Wiley Periodicals, Inc.

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

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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.

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

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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)

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

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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.

Mutations in PPIB (cyclophilin B) delay type I procollagen chain association and result in perinatal lethal to moderate osteogenesis imperfecta phenotypes.

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Pyott, Shawna M; Schwarze, Ulrike; Christiansen, Helena E; Pepin, Melanie G; Leistritz, Dru F; Dineen, Richard; Harris, Catharine; Burton, Barbara K; Angle, Brad; Kim, Katherine; Sussman, Michael D; Weis, Maryann; Eyre, David R; Russell, David W; McCarthy, Kevin J; Steiner, Robert D; Byers, Peter H

2011-04-15

Recessive mutations in the cartilage-associated protein (CRTAP), leucine proline-enriched proteoglycan 1 (LEPRE1) and peptidyl prolyl cis-trans isomerase B (PPIB) genes result in phenotypes that range from lethal in the perinatal period to severe deforming osteogenesis imperfecta (OI). These genes encode CRTAP (encoded by CRTAP), prolyl 3-hydroxylase 1 (P3H1; encoded by LEPRE1) and cyclophilin B (CYPB; encoded by PPIB), which reside in the rough endoplasmic reticulum (RER) and can form a complex involved in prolyl 3-hydroxylation in type I procollagen. CYPB, a prolyl cis-trans isomerase, has been thought to drive the prolyl-containing peptide bonds to the trans configuration needed for triple helix formation. Here, we describe mutations in PPIB identified in cells from three individuals with OI. Cultured dermal fibroblasts from the most severely affected infant make some overmodified type I procollagen molecules. Proα1(I) chains are slow to assemble into trimers, and abnormal procollagen molecules concentrate in the RER, and bind to protein disulfide isomerase (PDI) and prolyl 4-hydroxylase 1 (P4H1). These findings suggest that although CYPB plays a role in helix formation another effect is on folding of the C-terminal propeptide and trimer formation. The extent of procollagen accumulation and PDI/P4H1 binding differs among cells with mutations in PPIB, CRTAP and LEPRE1 with the greatest amount in PPIB-deficient cells and the least in LEPRE1-deficient cells. These findings suggest that prolyl cis-trans isomerase may be required to effectively fold the proline-rich regions of the C-terminal propeptide to allow proα chain association and suggest an order of action for CRTAP, P3H1 and CYPB in procollagen biosynthesis and pathogenesis of OI.

Histone deacetylase turnover and recovery in sulforaphane-treated colon cancer cells: competing actions of 14-3-3 and Pin1 in HDAC3/SMRT corepressor complex dissociation/reassembly

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Williams David E

2011-05-01

Full Text Available Abstract Background Histone deacetylase (HDAC inhibitors are currently undergoing clinical evaluation as anti-cancer agents. Dietary constituents share certain properties of HDAC inhibitor drugs, including the ability to induce global histone acetylation, turn-on epigenetically-silenced genes, and trigger cell cycle arrest, apoptosis, or differentiation in cancer cells. One such example is sulforaphane (SFN, an isothiocyanate derived from the glucosinolate precursor glucoraphanin, which is abundant in broccoli. Here, we examined the time-course and reversibility of SFN-induced HDAC changes in human colon cancer cells. Results Cells underwent progressive G2/M arrest over the period 6-72 h after SFN treatment, during which time HDAC activity increased in the vehicle-treated controls but not in SFN-treated cells. There was a time-dependent loss of class I and selected class II HDAC proteins, with HDAC3 depletion detected ahead of other HDACs. Mechanism studies revealed no apparent effect of calpain, proteasome, protease or caspase inhibitors, but HDAC3 was rescued by cycloheximide or actinomycin D treatment. Among the protein partners implicated in the HDAC3 turnover mechanism, silencing mediator for retinoid and thyroid hormone receptors (SMRT was phosphorylated in the nucleus within 6 h of SFN treatment, as was HDAC3 itself. Co-immunoprecipitation assays revealed SFN-induced dissociation of HDAC3/SMRT complexes coinciding with increased binding of HDAC3 to 14-3-3 and peptidyl-prolyl cis/trans isomerase 1 (Pin1. Pin1 knockdown blocked the SFN-induced loss of HDAC3. Finally, SFN treatment for 6 or 24 h followed by SFN removal from the culture media led to complete recovery of HDAC activity and HDAC protein expression, during which time cells were released from G2/M arrest. Conclusion The current investigation supports a model in which protein kinase CK2 phosphorylates SMRT and HDAC3 in the nucleus, resulting in dissociation of the corepressor

The Rab2A GTPase Promotes Breast Cancer Stem Cells and Tumorigenesis via Erk Signaling Activation

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Man-Li Luo

2015-04-01

Full Text Available Proline-directed phosphorylation is regulated by the prolyl isomerase Pin1, which plays a fundamental role in driving breast cancer stem-like cells (BCSCs. Rab2A is a small GTPase critical for vesicle trafficking. Here, we show that Pin1 increases Rab2A transcription to promote BCSC expansion and tumorigenesis in vitro and in vivo. Mechanistically, Rab2A directly interacts with and prevents dephosphorylation/inactivation of Erk1/2 by the MKP3 phosphatase, resulting in Zeb1 upregulation and β-catenin nuclear translocation. In cancer cells, Rab2A is activated via gene amplification, mutation or Pin1 overexpression. Rab2A overexpression or mutation endows BCSC traits to primary normal human breast epithelial cells, whereas silencing Rab2A potently inhibits the expansion and tumorigenesis of freshly isolated BCSCs. Finally, Rab2A overexpression correlates with poor clinical outcome in breast cancer patients. Thus, Pin1/Rab2A/Erk drives BCSC expansion and tumorigenicity, suggesting potential drug targets.

Lack of cyclophilin B in osteogenesis imperfecta with normal collagen folding.

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Barnes, Aileen M; Carter, Erin M; Cabral, Wayne A; Weis, MaryAnn; Chang, Weizhong; Makareeva, Elena; Leikin, Sergey; Rotimi, Charles N; Eyre, David R; Raggio, Cathleen L; Marini, Joan C

2010-02-11

Osteogenesis imperfecta is a heritable disorder that causes bone fragility. Mutations in type I collagen result in autosomal dominant osteogenesis imperfecta, whereas mutations in either of two components of the collagen prolyl 3-hydroxylation complex (cartilage-associated protein [CRTAP] and prolyl 3-hydroxylase 1 [P3H1]) cause autosomal recessive osteogenesis imperfecta with rhizomelia (shortening of proximal segments of upper and lower limbs) and delayed collagen folding. We identified two siblings who had recessive osteogenesis imperfecta without rhizomelia. They had a homozygous start-codon mutation in the peptidyl-prolyl isomerase B gene (PPIB), which results in a lack of cyclophilin B (CyPB), the third component of the complex. The proband's collagen had normal collagen folding and normal prolyl 3-hydroxylation, suggesting that CyPB is not the exclusive peptidyl-prolyl cis-trans isomerase that catalyzes the rate-limiting step in collagen folding, as is currently thought. 2010 Massachusetts Medical Society

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

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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.

Overexpression of PvPin1, a Bamboo Homolog of PIN1-Type Parvulin 1, Delays Flowering Time in Transgenic Arabidopsis and Rice

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Zhigang Zheng

2017-09-01

Full Text Available Because of the long and unpredictable flowering period in bamboo, the molecular mechanism of bamboo flowering is unclear. Recent study showed that Arabidopsis PIN1-type parvulin 1 (Pin1At is an important floral activator and regulates floral transition by facilitating the cis/trans isomerization of the phosphorylated Ser/Thr residues preceding proline motifs in suppressor of overexpression of CO 1 (SOC1 and agamous-like 24 (AGL24. Whether bamboo has a Pin1 homolog and whether it works in bamboo flowering are still unknown. In this study, we cloned PvPin1, a homolog of Pin1At, from Phyllostachys violascens (Bambusoideae. Bioinformatics analysis showed that PvPin1 is closely related to Pin1-like proteins in monocots. PvPin1 was widely expressed in all tested bamboo tissues, with the highest expression in young leaf and lowest in floral bud. Moreover, PvPin1 expression was high in leaves before bamboo flowering then declined during flower development. Overexpression of PvPin1 significantly delayed flowering time by downregulating SOC1 and AGL24 expression in Arabidopsis under greenhouse conditions and conferred a significantly late flowering phenotype by upregulating OsMADS56 in rice under field conditions. PvPin1 showed subcellular localization in both the nucleus and cytolemma. The 1500-bp sequence of the PvPin1 promoter was cloned, and cis-acting element prediction showed that ABRE and TGACG-motif elements, which responded to abscisic acid (ABA and methyl jasmonate (MeJA, respectively, were characteristic of P. violascens in comparison with Arabidopsis. On promoter activity analysis, exogenous ABA and MeJA could significantly inhibit PvPin1 expression. These findings suggested that PvPin1 may be a repressor in flowering, and its delay of flowering time could be regulated by ABA and MeJA in bamboo.

Overexpression of PvPin1, a Bamboo Homolog of PIN1-Type Parvulin 1, Delays Flowering Time in Transgenic Arabidopsis and Rice.

Science.gov (United States)

Zheng, Zhigang; Yang, Xiaoming; Fu, Yaping; Zhu, Longfei; Wei, Hantian; Lin, Xinchun

2017-01-01

Because of the long and unpredictable flowering period in bamboo, the molecular mechanism of bamboo flowering is unclear. Recent study showed that Arabidopsis PIN1-type parvulin 1 (Pin1At) is an important floral activator and regulates floral transition by facilitating the cis/trans isomerization of the phosphorylated Ser/Thr residues preceding proline motifs in suppressor of overexpression of CO 1 (SOC1) and agamous-like 24 (AGL24). Whether bamboo has a Pin1 homolog and whether it works in bamboo flowering are still unknown. In this study, we cloned PvPin1 , a homolog of Pin1At , from Phyllostachys violascens (Bambusoideae). Bioinformatics analysis showed that PvPin1 is closely related to Pin1-like proteins in monocots. PvPin1 was widely expressed in all tested bamboo tissues, with the highest expression in young leaf and lowest in floral bud. Moreover, PvPin1 expression was high in leaves before bamboo flowering then declined during flower development. Overexpression of PvPin1 significantly delayed flowering time by downregulating SOC1 and AGL24 expression in Arabidopsis under greenhouse conditions and conferred a significantly late flowering phenotype by upregulating OsMADS56 in rice under field conditions. PvPin1 showed subcellular localization in both the nucleus and cytolemma. The 1500-bp sequence of the PvPin1 promoter was cloned, and cis -acting element prediction showed that ABRE and TGACG-motif elements, which responded to abscisic acid (ABA) and methyl jasmonate (MeJA), respectively, were characteristic of P. violascens in comparison with Arabidopsis . On promoter activity analysis, exogenous ABA and MeJA could significantly inhibit PvPin1 expression. These findings suggested that PvPin1 may be a repressor in flowering, and its delay of flowering time could be regulated by ABA and MeJA in bamboo.

ORF Alignment: NC_005090 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... PEPTIDYL-PROLYL ISOMERASE [Wolinella succinogenes] ... Length = 147 ... Query: 29 ... VVVLETTSGTIELTLFPKAAPKAVENFTTH...VKNGYYDGLIFHRVIKRFMLQXXXXXXXXX 88 ... VVVLETTSGTIELTLFPKAAPKAVENFTTH...VKNGYYDGLIFHRVIKRFMLQ ... Sbjct: 1 ... VVVLETTSGTIELTLFPKAAPKAVENFTTHVKNGYYDGLIFHRVIKRFMLQGGDP

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...

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

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.

Arabidopsis Phosphomannose Isomerase 1, but Not Phosphomannose Isomerase 2, Is Essential for Ascorbic Acid Biosynthesis*S⃞

OpenAIRE

Maruta, Takanori; Yonemitsu, Miki; Yabuta, Yukinori; Tamoi, Masahiro; Ishikawa, Takahiro; Shigeoka, Shigeru

2008-01-01

We studied molecular and functional properties of Arabidopsis phosphomannose isomerase isoenzymes (PMI1 and PMI2) that catalyze reversible isomerization between d-fructose 6-phosphate and d-mannose 6-phosphate (Man-6P). The apparent Km and Vmax values for Man-6P of purified recombinant PMI1 were 41.3 ± 4.2 μm and 1.89 μmol/min/mg protein, respectively, whereas those of purified recombinant PMI2 were 372 ± 13 μm and 22.5 μmol/min/mg protein, respectively. Both PMI1 ...

Bioproduction of D-Tagatose from D-Galactose Using Phosphoglucose Isomerase from Pseudomonas aeruginosa PAO1.

Science.gov (United States)

Patel, Manisha J; Patel, Arti T; Akhani, Rekha; Dedania, Samir; Patel, Darshan H

2016-07-01

Pseudomonas aeruginosa PAO1 phosphoglucose isomerase was purified as an active soluble form by a single-step purification using Ni-NTA chromatography that showed homogeneity on SDS-PAGE with molecular mass ∼62 kDa. The optimum temperature and pH for the maximum isomerization activity with D-galactose were 60 °C and 7.0, respectively. Generally, sugar phosphate isomerases show metal-independent activity but PA-PGI exhibited metal-dependent isomerization activity with aldosugars and optimally catalyzed the D-galactose isomerization in the presence of 1.0 mM MnCl2. The apparent Km and Vmax for D-galactose under standardized conditions were calculated to be 1029 mM (±31.30 with S.E.) and 5.95 U/mg (±0.9 with S.E.), respectively. Equilibrium reached after 180 min with production of 567.51 μM D-tagatose from 1000 mM of D-galactose. Though, the bioconversion ratio is low but it can be increased by immobilization and enzyme engineering. Although various L-arabinose isomerases have been characterized for bioproduction of D-tagatose, P. aeruginosa glucose phosphate isomerase is distinguished from the other L-arabinose isomerases by its optimal temperature (60 °C) for D-tagatose production being mesophilic bacteria, making it an alternate choice for bulk production.

The PIN1 family gene PvPIN1 is involved in auxin-dependent root emergence and tillering in switchgrass

Directory of Open Access Journals (Sweden)

Kaijie Xu

2016-03-01

Full Text Available Abstract Switchgrass (Panicum virgatum L.; family Poaceae is a warm-season C4 perennial grass. Tillering plays an important role in determining the morphology of aboveground parts and the final biomass yield of switchgrass. Auxin distribution in plants can affect a variety of important growth and developmental processes, including the regulation of shoot and root branching, plant resistance and biological yield. Auxin transport and gradients in plants are mediated by influx and efflux carriers. PvPIN1, a switchgrass PIN1-like gene that is involved in regulating polar transport, is a putative auxin efflux carrier. Neighbor-joining analysis using sequences deposited in NCBI databases showed that the PvPIN1gene belongs to the PIN1 family and is evolutionarily closer to the Oryza sativa japonica group. Tiller emergence and development was significantly promoted in plants subjected toPvPIN1 RNA interference (RNAi, which yielded a phenotype similar to that of wild-type plants treated with the auxin transport inhibitor TIBA (2,3,5-triiodobenzoic acid. A transgenic approach that inducedPvPIN1 gene overexpression or suppression altered tiller number and the shoot/root ratio. These data suggest that PvPIN1plays an important role in auxin-dependent adventitious root emergence and tillering.

Fbw7α and Fbw7γ Collaborate To Shuttle Cyclin E1 into the Nucleolus for Multiubiquitylation

Science.gov (United States)

Bhaskaran, Nimesh; van Drogen, Frank; Ng, Hwee-Fang; Kumar, Raman; Ekholm-Reed, Susanna; Peter, Matthias

2013-01-01

Cyclin E1, an activator of cyclin-dependent kinase 2 (Cdk2) that promotes replicative functions, is normally expressed periodically within the mammalian cell cycle, peaking at the G1-S-phase transition. This periodicity is achieved by E2F-dependent transcription in late G1 and early S phases and by ubiquitin-mediated proteolysis. The ubiquitin ligase that targets phosphorylated cyclin E is SCFFbw7 (also known as SCFCdc4), a member of the cullin ring ligase (CRL) family. Fbw7, a substrate adaptor subunit, is expressed as three splice-variant isoforms with different subcellular distributions: Fbw7α is nucleoplasmic but excluded from the nucleolus, Fbw7β is cytoplasmic, and Fbw7γ is nucleolar. Degradation of cyclin E in vivo requires SCF complexes containing Fbw7α and Fbw7γ, respectively. In vitro reconstitution showed that the role of SCFFbw7α in cyclin E degradation, rather than ubiquitylation, is to serve as a cofactor of the prolyl cis-trans isomerase Pin1 in the isomerization of a noncanonical proline-proline bond in the cyclin E phosphodegron. This isomerization is required for subsequent binding and ubiquitylation by SCFFbw7γ. Here we show that Pin1-mediated isomerization of the cyclin E phosphodegron and subsequent binding to Fbw7γ drive nucleolar localization of cyclin E, where it is ubiquitylated by SCFFbw7γ prior to its degradation by the proteasome. It is possible that this constitutes a mechanism for rapid inactivation of phosphorylated cyclin E by nucleolar sequestration prior to its multiubiquitylation and degradation. PMID:23109421

Purification and characterization of the d-xylose isomerase gene from Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Ho, N W.Y.; Rosenfeld, S; Stevis, P; Tsao, G T

1983-11-01

A DNA fragment containing both the Escherichia coli D-xylose isomerase (D-xylose ketol-isomerase, EC 5.3.1.5) gene and the D-xylulokinase (ATP: D-xylulose 5-phosphotransferase, EC 2.7.1.17) gene has been cloned on an E. coli plasmid. The D-xylose isomerase gene was separated from the D-xylulokinase gene by the construction of a new deletion plasmid, pLX7. The D-xylose isomerase gene cloned on pLX7 was found still to be an intact gene. The precise location of the D-xylose isomerase gene on the plasmid pLX7 was further determined by the construction of two more plasmids, pLX8 and pLX9. This is believed to be the first D-xylose isomerase gene that has been isolated and extensively purified from any organism. D-Xylose isomerase, the enzyme product of the D-xylose isomerase gene, is responsible for the conversion of D-xylose to D-xylulose, as well as D-glucose to D-fructose. It is widely believed that yeast cannot ferment D-xylose to ethanol primarily because of the lack of D-xylose isomerase in yeast. D-Xylose isomerase (also known as D-glucose isomerase) is also used for the commercial production of high-fructose syrups. The purification of the D-xylose isomerase gene may lead to the following industrial applications: (1) cloning and expression of the gene in yeast to make the latter organism capable of directly fermenting D-xylose to ethanol, and (2) cloning of the gene on a high-copy-number plasmid in a proper host to overproduce the enzyme, which should have a profound impact on the high-fructose syrup technology. 14 references.

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

Solution structure of the parvulin-type PPIase domain of Staphylococcus aureus PrsA – Implications for the catalytic mechanism of parvulins

Directory of Open Access Journals (Sweden)

Koskela Harri

2009-03-01

Full Text Available Abstract Background Staphylococcus aureus is a Gram-positive pathogenic bacterium causing many kinds of infections from mild respiratory tract infections to life-threatening states as sepsis. Recent emergence of S. aureus strains resistant to numerous antibiotics has created a need for new antimicrobial agents and novel drug targets. S. aureus PrsA is a membrane associated extra-cytoplasmic lipoprotein which contains a parvulin-type peptidyl-prolyl cis-trans isomerase domain. PrsA is known to act as an essential folding factor for secreted proteins in Gram-positive bacteria and thus it is a potential target for antimicrobial drugs against S. aureus. Results We have solved a high-resolution solution structure of the parvulin-type peptidyl-prolyl cis-trans isomerase domain of S. aureus PrsA (PrsA-PPIase. The results of substrate peptide titrations pinpoint the active site and demonstrate the substrate preference of the enzyme. With detailed NMR spectroscopic investigation of the orientation and tautomeric state of the active site histidines we are able to give further insight into the structure of the catalytic site. NMR relaxation analysis gives information on the dynamic behaviour of PrsA-PPIase. Conclusion Detailed structural description of the S. aureus PrsA-PPIase lays the foundation for structure-based design of enzyme inhibitors. The structure resembles hPin1-type parvulins both structurally and regarding substrate preference. Even though a wealth of structural data is available on parvulins, the catalytic mechanism has yet to be resolved. The structure of S. aureus PrsA-PPIase and our findings on the role of the conserved active site histidines help in designing further experiments to solve the detailed catalytic mechanism.

Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90.

Science.gov (United States)

Blackburn, Elizabeth A; Wear, Martin A; Landré, Vivian; Narayan, Vikram; Ning, Jia; Erman, Burak; Ball, Kathryn L; Walkinshaw, Malcolm D

2015-09-01

Cyclophilin 40 (Cyp40) comprises an N-terminal cyclophilin domain with peptidyl-prolyl isomerase (PPIase) activity and a C-terminal tetratricopeptide repeat (TPR) domain that binds to the C-terminal-EEVD sequence common to both heat shock protein 70 (Hsp70) and Hsp90. We show in the present study that binding of peptides containing the MEEVD motif reduces the PPIase activity by ∼30%. CD and fluorescence assays show that the TPR domain is less stable than the cyclophilin domain and is stabilized by peptide binding. Isothermal titration calorimetry (ITC) shows that the affinity for the-MEEVD peptide is temperature sensitive in the physiological temperature range. Results from these biophysical studies fit with the MD simulations of the apo and holo (peptide-bound) structures which show a significant reduction in root mean square (RMS) fluctuation in both TPR and cyclophilin domains when-MEEVD is bound. The MD simulations of the apo-protein also highlight strong anti-correlated motions between residues around the PPIase-active site and a band of residues running across four of the seven helices in the TPR domain. Peptide binding leads to a distortion in the shape of the active site and a significant reduction in these strongly anti-correlated motions, providing an explanation for the allosteric effect of ligand binding and loss of PPIase activity. Together the experimental and MD results suggest that on heat shock, dissociation of Cyp40 from complexes mediated by the TPR domain leads to an increased pool of free Cyp40 capable of acting as an isomerase/chaperone in conditions of cellular stress. © 2015 Authors.

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):.

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.)

Triosephosphate isomerase is a common crystallization contaminant of soluble His-tagged proteins produced in Escherichia coli

International Nuclear Information System (INIS)

Kozlov, Guennadi; Vinaik, Roohi; Gehring, Kalle

2013-01-01

Crystals of E. coli triosephosphate isomerase were obtained as a contaminant and its structure was determined to 1.85 Å resolution. Attempts to crystallize several mammalian proteins overexpressed in Escherichia coli revealed a common contaminant, triosephosphate isomerase, a protein involved in glucose metabolism. Even with triosephosphate isomerase present in very small amounts, similarly shaped crystals appeared in the crystallization drops in a number of polyethylene glycol-containing conditions. All of the target proteins were His-tagged and their purification involved immobilized metal-affinity chromatography (IMAC), a step that was likely to lead to triosephosphate isomerase contamination. Analysis of the triosephosphate isomerase crystals led to the structure of E. coli triosephosphate isomerase at 1.85 Å resolution, which is a significant improvement over the previous structure

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.

Glucose (xylose) isomerase production from thermotolerant and ...

African Journals Online (AJOL)

Owner

2012-11-13

Nov 13, 2012 ... in the production of the high fructose corn syrup (HFCS) from corn starch. ... Key words: Glucose isomerase, xylose isomerase, enzyme activity, Klebsiella, ... Soil, water, and manure (five samples each) were collected from.

Phosphorylation of conserved PIN motifs directs Arabidopsis PIN1 polarity and auxin transport

NARCIS (Netherlands)

Huang, F.; Kemel Zago, M.; Abas, L.; van Marion, A.; Galván-Ampudia, C.S.; Offringa, R.

2010-01-01

Polar cell-to-cell transport of auxin by plasma membrane-localized PIN-FORMED (PIN) auxin efflux carriers generates auxin gradients that provide positional information for various plant developmental processes. The apical-basal polar localization of the PIN proteins that determines the direction of

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

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

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

Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidusd-arabinose isomerase

International Nuclear Information System (INIS)

Takeda, Kosei; Yoshida, Hiromi; Takada, Goro; Izumori, Ken; Kamitori, Shigehiro

2008-01-01

Recombinant B. pallidusd-arabinose isomerase was crystallized and diffraction data were collected to 2.3 Å resolution. d-Arabinose isomerase catalyzes the isomerization of d-arabinose to d-ribulose. Bacillus pallidusd-arabinose isomerase has broad substrate specificity and can catalyze the isomerization of d-arabinose, l-fucose, l-xylose, l-galactose and d-altrose. Recombinant B. pallidusd-arabinose isomerase was overexpressed, purified and crystallized. A crystal of the enzyme was obtained by the sitting-drop method at room temperature and belonged to the orthorhombic space group P2 1 2 1 2, with unit-cell parameters a = 144.9, b = 127.9, c = 109.5 Å. Diffraction data were collected to 2.3 Å resolution

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

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.

Functional differences in yeast protein disulfide isomerases

DEFF Research Database (Denmark)

Nørgaard, P; Westphal, V; Tachibana, C

2001-01-01

PDI1 is the essential gene encoding protein disulfide isomerase in yeast. The Saccharomyces cerevisiae genome, however, contains four other nonessential genes with homology to PDI1: MPD1, MPD2, EUG1, and EPS1. We have investigated the effects of simultaneous deletions of these genes. In several...

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.

Comparing the xylose reductase/xylitol dehydrogenase and xylose isomerase pathways in arabinose and xylose fermenting Saccharomyces cerevisiae strains

Directory of Open Access Journals (Sweden)

Hahn-Hägerdal Bärbel

2008-10-01

Full Text Available Abstract Background Ethanolic fermentation of lignocellulosic biomass is a sustainable option for the production of bioethanol. This process would greatly benefit from recombinant Saccharomyces cerevisiae strains also able to ferment, besides the hexose sugar fraction, the pentose sugars, arabinose and xylose. Different pathways can be introduced in S. cerevisiae to provide arabinose and xylose utilisation. In this study, the bacterial arabinose isomerase pathway was combined with two different xylose utilisation pathways: the xylose reductase/xylitol dehydrogenase and xylose isomerase pathways, respectively, in genetically identical strains. The strains were compared with respect to aerobic growth in arabinose and xylose batch culture and in anaerobic batch fermentation of a mixture of glucose, arabinose and xylose. Results The specific aerobic arabinose growth rate was identical, 0.03 h-1, for the xylose reductase/xylitol dehydrogenase and xylose isomerase strain. The xylose reductase/xylitol dehydrogenase strain displayed higher aerobic growth rate on xylose, 0.14 h-1, and higher specific xylose consumption rate in anaerobic batch fermentation, 0.09 g (g cells-1 h-1 than the xylose isomerase strain, which only reached 0.03 h-1 and 0.02 g (g cells-1h-1, respectively. Whereas the xylose reductase/xylitol dehydrogenase strain produced higher ethanol yield on total sugars, 0.23 g g-1 compared with 0.18 g g-1 for the xylose isomerase strain, the xylose isomerase strain achieved higher ethanol yield on consumed sugars, 0.41 g g-1 compared with 0.32 g g-1 for the xylose reductase/xylitol dehydrogenase strain. Anaerobic fermentation of a mixture of glucose, arabinose and xylose resulted in higher final ethanol concentration, 14.7 g l-1 for the xylose reductase/xylitol dehydrogenase strain compared with 11.8 g l-1 for the xylose isomerase strain, and in higher specific ethanol productivity, 0.024 g (g cells-1 h-1 compared with 0.01 g (g cells-1 h-1

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.

Immobilization of Recombinant Glucose Isomerase for Efficient Production of High Fructose Corn Syrup.

Science.gov (United States)

Jin, Li-Qun; Xu, Qi; Liu, Zhi-Qiang; Jia, Dong-Xu; Liao, Cheng-Jun; Chen, De-Shui; Zheng, Yu-Guo

2017-09-01

Glucose isomerase is the important enzyme for the production of high fructose corn syrup (HFCS). One-step production of HFCS containing more than 55% fructose (HFCS-55) is receiving much attention for its industrial applications. In this work, the Escherichia coli harboring glucose isomerase mutant TEGI-W139F/V186T was immobilized for efficient production of HFCS-55. The immobilization conditions were optimized, and the maximum enzyme activity recovery of 92% was obtained. The immobilized glucose isomerase showed higher pH, temperature, and operational stabilities with a K m value of 272 mM and maximum reaction rate of 23.8 mM min -1 . The fructose concentration still retained above 55% after the immobilized glucose isomerase was reused for 10 cycles, and more than 85% of its initial activity was reserved even after 15 recycles of usage at temperature of 90 °C. The results highlighted the immobilized glucose isomerase as a potential biocatalyst for HFCS-55 production.

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

Postirradiation examinations of fuel pins from the GCFR F-1 series of mixed-oxide fuel pins at 5.5 at. % burnup

International Nuclear Information System (INIS)

Strain, R.V.; Johnson, C.E.

1978-05-01

Postirradiation examinations were performed on five fuel pins from the Gas-Cooled Fast-Breeder Reactor F-1 experiment irradiated in EBR-II to a peak burnup of approximately 5.5 at. %. These encapsulated fuel pins were irradiated at peak-power linear ratings from approximately 13 to 15 kW/ft and peak cladding inside diameter temperatures from approximately 625 to 760 0 C. The maximum diametral change that occurred during irradiation was 0.2% ΔD/D 0 . The maximum fuel-cladding chemical interaction depth was 2.6 mils in fuel pin G-1 and 1 mil or less in the other three pins examined destructively. Significant migration of the volatile fission products occurred axially to the fuel-blanket interfaces. Teh postirradiation examination data indicate that fuel melted at the inner surface of the annular fuel pellets in the two highest power rating fuel pins, but little axial movement of fuel occurred

Postirradiation examinations of fuel pins from the GCFR F-1 series of mixed-oxide fuel pins at 5. 5 at. % burnup

Energy Technology Data Exchange (ETDEWEB)

Strain, R V; Johnson, C E

1978-05-01

Postirradiation examinations were performed on five fuel pins from the Gas-Cooled Fast-Breeder Reactor F-1 experiment irradiated in EBR-II to a peak burnup of approximately 5.5 at. %. These encapsulated fuel pins were irradiated at peak-power linear ratings from approximately 13 to 15 kW/ft and peak cladding inside diameter temperatures from approximately 625 to 760/sup 0/C. The maximum diametral change that occurred during irradiation was 0.2% ..delta..D/D/sub 0/. The maximum fuel-cladding chemical interaction depth was 2.6 mils in fuel pin G-1 and 1 mil or less in the other three pins examined destructively. Significant migration of the volatile fission products occurred axially to the fuel-blanket interfaces. Teh postirradiation examination data indicate that fuel melted at the inner surface of the annular fuel pellets in the two highest power rating fuel pins, but little axial movement of fuel occurred.

Structure of ribose 5-phosphate isomerase from the probiotic bacterium Lactobacillus salivarius UCC118

International Nuclear Information System (INIS)

Lobley, Carina M. C.; Aller, Pierre; Douangamath, Alice; Reddivari, Yamini; Bumann, Mario; Bird, Louise E.; Nettleship, Joanne E.; Brandao-Neto, Jose; Owens, Raymond J.; O’Toole, Paul W.; Walsh, Martin A.

2012-01-01

The crystal structure of ribose 5-phosphate isomerase has been determined to 1.72 Å resolution and is presented with a brief comparison to other known ribose 5-phosphate isomerase A structures. The structure of ribose 5-phosphate isomerase from the probiotic bacterium Lactobacillus salivarius UCC188 has been determined at 1.72 Å resolution. The structure was solved by molecular replacement, which identified the functional homodimer in the asymmetric unit. Despite only showing 57% sequence identity to its closest homologue, the structure adopted the typical α and β d-ribose 5-phosphate isomerase fold. Comparison to other related structures revealed high homology in the active site, allowing a model of the substrate-bound protein to be proposed. The determination of the structure was expedited by the use of in situ crystallization-plate screening on beamline I04-1 at Diamond Light Source to identify well diffracting protein crystals prior to routine cryocrystallography

FKBP immunophilins and Alzheimer's disease: A chaperoned affair

Indian Academy of Sciences (India)

All FKBPs contain a domain with prolyl cis/trans isomerase (PPIase) activity. ... FKBP proteins are expressed ubiquitously but show relatively high levels of expression in the nervous .... evolution, higher organisms like humans, who normally.

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.

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.

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.

Escherichia coli rpiA gene encoding ribose phosphate isomerase A

DEFF Research Database (Denmark)

Hove-Jensen, Bjarne; Maigaard, Marianne

1993-01-01

The rpiA gene encoding ribose phosphate isomerase A was cloned from phage 1A2(471) of the Kohara gene library. Subcloning, restriction, and complementation analyses revealed an 1,800-bp SspI-generated DNA fragment that contained the entire control and coding sequences. This DNA fragment was seque......The rpiA gene encoding ribose phosphate isomerase A was cloned from phage 1A2(471) of the Kohara gene library. Subcloning, restriction, and complementation analyses revealed an 1,800-bp SspI-generated DNA fragment that contained the entire control and coding sequences. This DNA fragment...

Development of a fast pin-by-pin transport solver in ARCADIA registered

International Nuclear Information System (INIS)

Geemert, R. van

2009-01-01

For satisfaction of future global customer needs, dedicated efforts are being coordinated internationally and pursued continuously at AREVA NP. The currently ongoing CONVERGENCE project is committed to the development of the ARCADIA registered next generation core simulation software package. ARCADIA registered will be put to global use by all AREVA NP business regions, for the entire spectrum of core design processes, licensing computations and safety studies. As part of the currently ongoing trend towards more sophisticated neutronics methodologies, an SP 3 nodal transport concept (van Geemert 2008) has been developed for ARTEMIS (Hobson 2008) which is the steady-state and transient core simulation part of ARCADIA registered . For enabling a high computational performance, the SP 3 calculations are accelerated by applying multi-level coarse mesh rebalancing (van Geemert 2006). In the current implementation, SP 3 is typically about 1.4 times as expensive computationally as SP 1 (diffusion). The developed SP 3 solution concept is foreseen as the future computational workhorse for many-group 3D pin-by-pin full core computations by ARCADIA registered . With the entire numerical workload being highly parallelizable through domain decomposition techniques, associated CPU-time requirements that adhere to the efficiency needs in the nuclear industry can be expected to become feasible in the near future. The accuracy enhancement obtainable by using SP 3 instead of SP 1 has been verified by a detailed comparison of ARTEMIS 16-group pin-by-pin SP N results with KAERI's DeCart reference results (Kozlowski 2003) for the 2D pin-by-pin Purdue UO 2 /MOX benchmark. Within the associated pin-by-pin grid, large pin-to-pin variations in cross-section values occur due to the explicit modelling of guide tubes, gadolinium pins as well as the heterogeneous distribution of MOX assemblies and UO 2 assemblies featuring significantly different burnups. With a pin-by-pin grid as

Characterization of a mutated Geobacillus stearothermophilus L-arabinose isomerase that increases the production rate of D-tagatose.

Science.gov (United States)

Kim, H-J; Kim, J-H; Oh, H-J; Oh, D-K

2006-07-01

Characterization of a mutated Geobacillus stearothermophilus L-arabinose isomerase used to increase the production rate of D-tagatose. A mutated gene was obtained by an error-prone polymerase chain reaction using L-arabinose isomerase gene from G. stearothermophilus as a template and the gene was expressed in Escherichia coli. The expressed mutated L-arabinose isomerase exhibited the change of three amino acids (Met322-->Val, Ser393-->Thr, and Val408-->Ala), compared with the wild-type enzyme and was then purified to homogeneity. The mutated enzyme had a maximum galactose isomerization activity at pH 8.0, 65 degrees C, and 1.0 mM Co2+, while the wild-type enzyme had a maximum activity at pH 8.0, 60 degrees C, and 1.0-mM Mn2+. The mutated L-arabinose isomerase exhibited increases in D-galactose isomerization activity, optimum temperature, catalytic efficiency (kcat/Km) for D-galactose, and the production rate of D-tagatose from D-galactose. The mutated L-arabinose isomerase from G. stearothermophilus is valuable for the commercial production of D-tagatose. This work contributes knowledge on the characterization of a mutated L-arabinose isomerase, and allows an increased production rate for D-tagatose from D-galactose using the mutated enzyme.

A single and two step isomerization process for d-tagatose and l-ribose bioproduction using l-arabinose isomerase and d-lyxose isomerase.

Science.gov (United States)

Patel, Manisha J; Akhani, Rekha C; Patel, Arti T; Dedania, Samir R; Patel, Darshan H

2017-02-01

l-ribose and d-tagatose are biochemically synthesized using sugar isomerases. The l-arabinose isomerase gene from Shigella flexneri (Sf-AI) was cloned and expressed in Escherichia coli BL-21. Sf-AI was applied for the bioproduction of d-tagatose from d-galactose. l-ribose synthesis was performed by two step isomerization using Sf-AI and d-lyxose/ribose isomerase from Cohnella laevoribosii. The overall 22.3% and 25% conversion rate were observed for d-tagatose and l-ribose production from d-galactose and l-arabinose respectively. In the present manuscript, synthesis of rare sugars from naturally available sugars is discussed along with the biochemical characterization of Sf-AI and its efficiency. Copyright © 2016 Elsevier Inc. All rights reserved.

Crystallization and preliminary X-ray characterization of phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv

International Nuclear Information System (INIS)

Mathur, Divya; Anand, Kanchan; Mathur, Deepika; Jagadish, Nirmala; Suri, Anil; Garg, Lalit C.

2007-01-01

The phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv was crystallized and diffraction data were collected to 2.8 Å resolution. Phosphoglucose isomerase is a ubiquitous enzyme that catalyzes the isomerization of d-glucopyranose-6-phosphate to d-fructofuranose-6-phosphate. The present investigation reports the expression, purification, crystallization and preliminary crystallographic studies of the phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv, which shares 46% sequence identity with that of its human host. The recombinant protein, which was prepared using an Escherichia coli expression system, was crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.8 Å and belonged to the orthorhombic space group I2 1 2 1 2 1 , with unit-cell parameters a = 109.0, b = 119.8, c = 138.9 Å

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

Proč axony nezabloudí

Czech Academy of Sciences Publication Activity Database

Balaštík, Martin

2017-01-01

Roč. 96, č. 2 (2017), s. 78-79 ISSN 0042-4544 R&D Projects: GA ČR(CZ) GA16-15915S Institutional support: RVO:67985823 Keywords : axon guidance * prolyl isomerase * neural development Subject RIV: FH - Neurology OBOR OECD: Developmental biology

Pinning, de-pinning and re-pinning of a slowly varying rivulet

KAUST Repository

Paterson, C.; Wilson, S.K.; Duffy, B.R.

2013-01-01

The solutions for the unidirectional flow of a thin rivulet with prescribed volume flux down an inclined planar substrate are used to describe the locally unidirectional flow of a rivulet with constant width (i.e. pinned contact lines) but slowly varying contact angle as well as the possible pinning and subsequent de-pinning of a rivulet with constant contact angle and the possible de-pinning and subsequent re-pinning of a rivulet with constant width as they flow in the azimuthal direction from the top to the bottom of a large horizontal cylinder. Despite being the same locally, the global behaviour of a rivulet with constant width can be very different from that of a rivulet with constant contact angle. In particular, while a rivulet with constant non-zero contact angle can always run from the top to the bottom of the cylinder, the behaviour of a rivulet with constant width depends on the value of the width. Specifically, while a narrow rivulet can run all the way from the top to the bottom of the cylinder, a wide rivulet can run from the top of the cylinder only to a critical azimuthal angle. The scenario in which the hitherto pinned contact lines of the rivulet de-pin at the critical azimuthal angle and the rivulet runs from the critical azimuthal angle to the bottom of the cylinder with zero contact angle but slowly varying width is discussed. The pinning and de-pinning of a rivulet with constant contact angle, and the corresponding situation involving the de-pinning and re-pinning of a rivulet with constant width at a non-zero contact angle which generalises the de-pinning at zero contact angle discussed earlier, are described. In the latter situation, the mass of fluid on the cylinder is found to be a monotonically increasing function of the constant width. © 2013 Elsevier Masson SAS. All rights reserved.

Pinning, de-pinning and re-pinning of a slowly varying rivulet

KAUST Repository

Paterson, C.

2013-09-01

The solutions for the unidirectional flow of a thin rivulet with prescribed volume flux down an inclined planar substrate are used to describe the locally unidirectional flow of a rivulet with constant width (i.e. pinned contact lines) but slowly varying contact angle as well as the possible pinning and subsequent de-pinning of a rivulet with constant contact angle and the possible de-pinning and subsequent re-pinning of a rivulet with constant width as they flow in the azimuthal direction from the top to the bottom of a large horizontal cylinder. Despite being the same locally, the global behaviour of a rivulet with constant width can be very different from that of a rivulet with constant contact angle. In particular, while a rivulet with constant non-zero contact angle can always run from the top to the bottom of the cylinder, the behaviour of a rivulet with constant width depends on the value of the width. Specifically, while a narrow rivulet can run all the way from the top to the bottom of the cylinder, a wide rivulet can run from the top of the cylinder only to a critical azimuthal angle. The scenario in which the hitherto pinned contact lines of the rivulet de-pin at the critical azimuthal angle and the rivulet runs from the critical azimuthal angle to the bottom of the cylinder with zero contact angle but slowly varying width is discussed. The pinning and de-pinning of a rivulet with constant contact angle, and the corresponding situation involving the de-pinning and re-pinning of a rivulet with constant width at a non-zero contact angle which generalises the de-pinning at zero contact angle discussed earlier, are described. In the latter situation, the mass of fluid on the cylinder is found to be a monotonically increasing function of the constant width. © 2013 Elsevier Masson SAS. All rights reserved.

ZIFL1.1 transporter modulates polar auxin transport by stabilizing membrane abundance of multiple PINs in Arabidopsis root tip

Science.gov (United States)

Remy, Estelle; Baster, Pawel; Friml, Jiří; Duque, Paula

2013-01-01

Cell-to-cell directional flow of the phytohormone auxin is primarily established by polar localization of the PIN auxin transporters, a process tightly regulated at multiple levels by auxin itself. We recently reported that, in the context of strong auxin flows, activity of the vacuolar ZIFL1.1 transporter is required for fine-tuning of polar auxin transport rates in the Arabidopsis root. In particular, ZIFL1.1 function protects plasma-membrane stability of the PIN2 carrier in epidermal root tip cells under conditions normally triggering PIN2 degradation. Here, we show that ZIFL1.1 activity at the root tip also promotes PIN1 plasma-membrane abundance in central cylinder cells, thus supporting the notion that ZIFL1.1 acts as a general positive modulator of polar auxin transport in roots. PMID:23857365

Reduction of halo pin site morbidity with a new pin care regimen.

Science.gov (United States)

Kazi, Hussain Anthony; de Matas, Marcus; Pillay, Robin

2013-06-01

A retrospective analysis of halo device associated morbidity over a 4-year period. To assess the impact of a new pin care regimen on halo pin site related morbidity. Halo orthosis treatment still has a role in cervical spine pathology, despite increasing possibilities of open surgical treatment. Published figures for pin site infection range from 12% to 22% with pin loosening from 7% to 50%. We assessed the outcome of a new pin care regimen on morbidity associated with halo spinal orthoses, using a retrospective cohort study from 2001 to 2004. In the last two years, our pin care regimen was changed. This involved pin site care using chlorhexidene & regular torque checking as part of a standard protocol. Previously, povidone iodine was used as skin preparation in theatre, followed by regular sterile saline cleansing when pin sites became encrusted with blood. There were 37 patients in the series, the median age was 49 (range, 22-83) and 20 patients were male. The overall infection rate prior to the new pin care protocol was 30% (n=6) and after the introduction, it dropped to 5.9% (n=1). This difference was statistically significant (p<0.05). Pin loosening occurred in one patient in the group prior to the formal pin care protocol (3%) and none thereafter. Reduced morbidity from halo use can be achieved with a modified pin cleansing and tightening regimen.

Directed evolution of xylose isomerase for improved xylose catabolism and fermentation in the yeast Saccharomyces cerevisiae.

Science.gov (United States)

Lee, Sun-Mi; Jellison, Taylor; Alper, Hal S

2012-08-01

The heterologous expression of a highly functional xylose isomerase pathway in Saccharomyces cerevisiae would have significant advantages for ethanol yield, since the pathway bypasses cofactor requirements found in the traditionally used oxidoreductase pathways. However, nearly all reported xylose isomerase-based pathways in S. cerevisiae suffer from poor ethanol productivity, low xylose consumption rates, and poor cell growth compared with an oxidoreductase pathway and, additionally, often require adaptive strain evolution. Here, we report on the directed evolution of the Piromyces sp. xylose isomerase (encoded by xylA) for use in yeast. After three rounds of mutagenesis and growth-based screening, we isolated a variant containing six mutations (E15D, E114G, E129D, T142S, A177T, and V433I) that exhibited a 77% increase in enzymatic activity. When expressed in a minimally engineered yeast host containing a gre3 knockout and tal1 and XKS1 overexpression, the strain expressing this mutant enzyme improved its aerobic growth rate by 61-fold and both ethanol production and xylose consumption rates by nearly 8-fold. Moreover, the mutant enzyme enabled ethanol production by these yeasts under oxygen-limited fermentation conditions, unlike the wild-type enzyme. Under microaerobic conditions, the ethanol production rates of the strain expressing the mutant xylose isomerase were considerably higher than previously reported values for yeast harboring a xylose isomerase pathway and were also comparable to those of the strains harboring an oxidoreductase pathway. Consequently, this study shows the potential to evolve a xylose isomerase pathway for more efficient xylose utilization.

High production of D-tagatose, a potential sugar substitute, using immobilized L-arabinose isomerase.

Science.gov (United States)

Kim, P; Yoon, S H; Roh, H J; Choi, J H

2001-01-01

An L-arabinose isomerase of Escherichia coli was immobilized using covalent binding to agarose to produce D-tagatose, a bulking sweetener that can be economically used as a sugar substitute. The immobilized L-arabinose isomerase stably produced an average of 7.5 g-tagatose/L.day for 7 days with a productivity exceeding that of the free enzyme (0.47 vs 0.30 mg/U.day). Using a scaled-up immobilized enzyme system, 99.9 g-tagatose/L was produced from galactose with 20% equilibrium in 48 h. The process was repeated two more times with production of 104.1 and 103.5 g-tagatose/L. D-Tagatose production using an immobilized L-arabinose isomerase has a high potential for commercial application.

Domain architecture of protein-disulfide isomerase facilitates its dual role as an oxidase and an isomerase in Ero1p-mediated disulfide formation

DEFF Research Database (Denmark)

Kulp, M. S.; Frickel, E. M.; Ellgaard, Lars

2006-01-01

reduction/rearrangement of non-native disulfides is poorly understood. We analyzed the role of individual PDI domains in disulfide bond formation in a reaction driven by their natural oxidant, Ero1p. We found that Ero1p oxidizes the isolated PDI catalytic thioredoxin domains, A and A' at the same rate......Native disulfide bond formation in eukaryotes is dependent on protein-disulfide isomerase (PDI) and its homologs, which contain varying combinations of catalytically active and inactive thioredoxin domains. However, the specific contribution of PDI to the formation of new disulfides versus...... catalytic (A) domain. The specific order of thioredoxin domains in PDI is important in establishing the asymmetry in the rate of oxidation of the two active sites thus allowing A and A', two thioredoxin domains that are similar in sequence and structure, to serve opposing functional roles as a disulfide...

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

Biochemical function of typical and variant Arabidopsis thaliana U-box E3 ubiquitin-protein ligases

DEFF Research Database (Denmark)

Wiborg, Jakob; O'Shea, Charlotte; Skriver, Karen

2008-01-01

of the distant U-box protein, AtPUB49, representing a large family of eukaryotic proteins containing a U-box linked to a cyclophilin-like peptidyl-prolyl cis-trans isomerase domain, was characterized biochemically. AtPUB49 functioned both as a prolyl isomerase and a chaperone by catalysing cis......The variance of the U-box domain in 64 Arabidopsis thaliana (thale cress) E3s (ubiquitin-protein ligases) was used to examine the interactions between E3s and E2s (ubiquitin-conjugating enzymes). E2s and E3s are components of the ubiquitin protein degradation pathway. Seven U-box proteins were...... analysed for their ability to ubiquitinate proteins in vitro in co-operation with different E2s. All U-box domains exhibited ubiquitination activity and interacted productively with UBC4/5-type E2s. Three and four of the U-box domains mediated ubiquitin addition in the presence of UBC13 and UBC7 E2s...

PPIB mutations cause severe osteogenesis imperfecta.

Science.gov (United States)

van Dijk, Fleur S; Nesbitt, Isabel M; Zwikstra, Eline H; Nikkels, Peter G J; Piersma, Sander R; Fratantoni, Silvina A; Jimenez, Connie R; Huizer, Margriet; Morsman, Alice C; Cobben, Jan M; van Roij, Mirjam H H; Elting, Mariet W; Verbeke, Jonathan I M L; Wijnaendts, Liliane C D; Shaw, Nick J; Högler, Wolfgang; McKeown, Carole; Sistermans, Erik A; Dalton, Ann; Meijers-Heijboer, Hanne; Pals, Gerard

2009-10-01

Deficiency of cartilage-associated protein (CRTAP) or prolyl 3-hydroxylase 1(P3H1) has been reported in autosomal-recessive lethal or severe osteogenesis imperfecta (OI). CRTAP, P3H1, and cyclophilin B (CyPB) form an intracellular collagen-modifying complex that 3-hydroxylates proline at position 986 (P986) in the alpha1 chains of collagen type I. This 3-prolyl hydroxylation is decreased in patients with CRTAP and P3H1 deficiency. It was suspected that mutations in the PPIB gene encoding CyPB would also cause OI with decreased collagen 3-prolyl hydroxylation. To our knowledge we present the first two families with recessive OI caused by PPIB gene mutations. The clinical phenotype is compatible with OI Sillence type II-B/III as seen with COL1A1/2, CRTAP, and LEPRE1 mutations. The percentage of 3-hydroxylated P986 residues in patients with PPIB mutations is decreased in comparison to normal, but it is higher than in patients with CRTAP and LEPRE1 mutations. This result and the fact that CyPB is demonstrable independent of CRTAP and P3H1, along with reported decreased 3-prolyl hydroxylation due to deficiency of CRTAP lacking the catalytic hydroxylation domain and the known function of CyPB as a cis-trans isomerase, suggest that recessive OI is caused by a dysfunctional P3H1/CRTAP/CyPB complex rather than by the lack of 3-prolyl hydroxylation of a single proline residue in the alpha1 chains of collagen type I.

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

OpenAIRE

Daar, I O; Artymiuk, P J; Phillips, D C; Maquat, L E

1986-01-01

Triose-phosphate isomerase (TPI; D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1) deficiency is a recessive disorder that results in hemolytic anemia and neuromuscular dysfunction. To determine the molecular basis of this disorder, a TPI allele from two unrelated patients homozygous for TPI deficiency was compared with an allele from a normal individual. Each disease-associated sequence harbors a G X C----C X G transversion in the codon for amino acid-104 and specifies a structurally...

Thermal-Hydraulic Simulations of Single Pin and Assembly Sector for IVG- 1M Reactor

Energy Technology Data Exchange (ETDEWEB)

Kraus, A. [Argonne National Lab. (ANL), Argonne, IL (United States); Garner, P. [Argonne National Lab. (ANL), Argonne, IL (United States); Hanan, N. [Argonne National Lab. (ANL), Argonne, IL (United States)

2015-01-15

Thermal-hydraulic simulations have been performed using computational fluid dynamics (CFD) for the highly-enriched uranium (HEU) design of the IVG.1M reactor at the Institute of Atomic Energy (IAE) at the National Nuclear Center (NNC) in the Republic of Kazakhstan. Steady-state simulations were performed for both types of fuel assembly (FA), i.e. the FA in rows 1 & 2 and the FA in row 3, as well as for single pins in those FA (600 mm and 800 mm pins). Both single pin calculations and bundle sectors have been simulated for the most conservative operating conditions corresponding to the 10 MW output power, which corresponds to a pin unit cell Reynolds number of only about 7500. Simulations were performed using the commercial code STAR-CCM+ for the actual twisted pin geometry as well as a straight-pin approximation. Various Reynolds-Averaged Navier-Stokes (RANS) turbulence models gave different results, and so some validation runs with a higher-fidelity Large Eddy Simulation (LES) code were performed given the lack of experimental data. These singled out the Realizable Two-Layer k-ε as the most accurate turbulence model for estimating surface temperature. Single-pin results for the twisted case, based on the average flow rate per pin and peak pin power, were conservative for peak clad surface temperature compared to the bundle results. Also the straight-pin calculations were conservative as compared to the twisted pin simulations, as expected, but the single-pin straight case was not always conservative with regard to the straight-pin bundle. This was due to the straight-pin temperature distribution being strongly influenced by the pin orientation, particularly near the outer boundary. The straight-pin case also predicted the peak temperature to be in a different location than the twisted-pin case. This is a limitation of the straight-pin approach. The peak temperature pin was in a different location from the peak power pin in every case simulated, and occurred at an

Effect of gamma irradiation on whole-cell glucose isomerase. Pt.1

International Nuclear Information System (INIS)

Bachman, S.; Gebicka, L.

1984-01-01

Gamma-rays induced inactivation of Actinoplanes missouriensis and Streptomyces olivaceus glucose isomerase has been studied. This enzyme exhibits high resistance against ionizing radiation. The D 37 value was found to be equal to 131 kGy for Actinoplanes missouriensis cells and 88 kGy for Streptomyces olivaceus cells when irradiated in the dry state in the presence of air. Mg 2+ ions do not affect the radiosensitivity of the enzyme in cells, while the addition of Co 2+ ions to the cell suspension increases its stability against ionizing radiation. (orig.) [de

Neurological findings in triosephosphate isomerase deficiency

NARCIS (Netherlands)

Poll-The, B. T.; Aicardi, J.; Girot, R.; Rosa, R.

1985-01-01

Two siblings with hemolytic anemia caused by triosephosphate isomerase deficiency developed a progressive neurological syndrome featuring dystonic movements, tremor, pyramidal tract signs, and evidence of spinal motor neuron involvement. Intelligence was unaffected. The findings in these patients

Ricinus communis cyclophilin: functional characterisation of a sieve tube protein involved in protein folding.

Science.gov (United States)

Gottschalk, Maren; Dolgener, Elmar; Xoconostle-Cázares, Beatriz; Lucas, William J; Komor, Ewald; Schobert, Christian

2008-09-01

The phloem translocation stream of the angiosperms contains a special population of proteins and RNA molecules which appear to be produced in the companion cells prior to being transported into the sieve tube system through the interconnecting plasmodesmata. During this process, these non-cell-autonomous proteins are thought to undergo partial unfolding. Recent mass spectroscopy studies identified peptidyl-prolyl cis-trans isomerase (PPIases) as potential molecular chaperones functioning in the phloem translocation stream (Giavalisco et al. 2006). In the present study, we describe the cloning and characterisation of a castor bean phloem cyclophilin, RcCYP1 that has high peptidyl-prolyl cis-trans isomerase activity. Equivalent enzymatic activity was detected with phloem sap or purified recombinant (His)(6)-tagged RcCYP1. Mass spectrometry analysis of proteolytic peptides, derived from a 22 kDa band in HPLC-fractionated phloem sap, immunolocalisation studies and Western analysis of proteins extracted from castor bean tissues/organs indicated that RcCYP1 is an abundant protein in the companion cell-sieve element complex. Microinjection experiments established that purified recombinant (His)(6)-RcCYP1 can interact with plasmodesmata to both induce an increase in size exclusion limit and mediate its own cell-to-cell trafficking. Collectively, these findings support the hypothesis that RcCYP1 plays a role in the refolding of non-cell-autonomous proteins after their entry into the phloem translocation stream.

Computer simulation of vortex pinning in type II superconductors. II. Random point pins

International Nuclear Information System (INIS)

Brandt, E.H.

1983-01-01

Pinning of vortices in a type II superconductor by randomly positioned identical point pins is simulated using the two-dimensional method described in a previous paper (Part I). The system is characterized by the vortex and pin numbers (N/sub v/, N/sub p/), the vortex and pin interaction ranges (R/sub v/, R/sub p/), and the amplitude of the pin potential A/sub p/. The computation is performed for many cases: dilute or dense, sharp or soft, attractive or repulsive, weak or strong pins, and ideal or amorphous vortex lattice. The total pinning force F as a function of the mean vortex displacment X increases first linearly (over a distance usually much smaller than the vortex spacing and than R/sub p/) and then saturates, fluctuating about its averaging F-bar. We interpret F-bar as the maximum pinning force j/sub c/B of a large specimen. For weak pins the prediction of Larkin and Ovchinnikov for two-dimensional collective pinning is confirmed: F-bar = const. iW/R/sub p/c 66 , where W-bar is the mean square pinning force and c 66 is the shear modulus of the vortex lattice. If the initial vortex lattice is chosen highly defective (''amorphous'') the constant is 1.3--3 times larger than for the ideal triangular lattice. This finding may explain the often observed ''history effect.'' The function F-bar(A/sub p/) exhibits a jump, which for dilute, sharp, attractive pins occurs close to the ''threshold value'' predicted for isolated pins by Labusch. This jump reflects the onset of plastic deformation of the vortex lattice, and in some cases of vortex trapping, but is not a genuine threshold

The Role of Exportin-5 in MicroRNA Biogenesis and Cancer

Directory of Open Access Journals (Sweden)

Ke Wu

2018-04-01

Full Text Available MicroRNAs (miRNAs are conserved small non-coding RNAs that play an important role in the regulation of gene expression and participate in a variety of biological processes. The biogenesis of miRNAs is tightly controlled at multiple steps, such as transcription of miRNA genes, processing by Drosha and Dicer, and transportation of precursor miRNAs (pre-miRNAs from the nucleus to the cytoplasm by exportin-5 (XPO5. Given the critical role of nuclear export of pre-miRNAs in miRNA biogenesis, any alterations of XPO5, resulting from either genetic mutation, epigenetic change, abnormal expression level or posttranslational modification, could affect miRNA expression and thus have profound effects on tumorigenesis. Importantly, XPO5 phosphorylation by ERK kinase and its cis/trans isomerization by the prolyl isomerase Pin1 impair XPO5′s nucleo-to-cytoplasmic transport ability of pre-miRNAs, leading to downregulation of mature miRNAs in hepatocellular carcinoma. In this review, we focus on how XPO5 transports pre-miRNAs in the cells and summarize the dysregulation of XPO5 in human tumors. Keywords: Exportin-5, MicroRNA, Biogenesis, Dysregulation, Cancer

Mapping the dynamics of ligand reorganization via {sup 13}CH{sub 3} and {sup 13}CH{sub 2} relaxation dispersion at natural abundance

Energy Technology Data Exchange (ETDEWEB)

Peng, Jeffrey W., E-mail: jpeng@nd.edu; Wilson, Brian D.; Namanja, Andrew T. [University of Notre Dame, Department of Chemistry and Biochemistry (United States)

2009-09-15

Flexible ligands pose challenges to standard structure-activity studies since they frequently reorganize their conformations upon protein binding and catalysis. Here, we demonstrate the utility of side chain {sup 13}C relaxation dispersion measurements to identify and quantify the conformational dynamics that drive this reorganization. The dispersion measurements probe methylene {sup 13}CH{sub 2} and methyl {sup 13}CH{sub 3} groups; the latter are highly prevalent side chain moieties in known drugs. Combining these side chain studies with existing backbone dispersion studies enables a comprehensive investigation of {mu}s-ms conformational dynamics related to binding and catalysis. We perform these measurements at natural {sup 13}C abundance, in congruence with common pharmaceutical research settings. We illustrate these methods through a study of the interaction of a phosphopeptide ligand with the peptidyl-prolyl isomerase, Pin1. The results illuminate the side-chain moieties that undergo conformational readjustments upon complex formation. In particular, we find evidence that multiple exchange processes influence the side chain dispersion profiles. Collectively, our studies illustrate how side-chain relaxation dispersion can shed light on ligand conformational transitions required for activity, and thereby suggest strategies for its optimization.

PINS Spectrum Identification Guide

Energy Technology Data Exchange (ETDEWEB)

A.J. Caffrey

2012-03-01

The Portable Isotopic Neutron Spectroscopy—PINS, for short—system identifies the chemicals inside munitions and containers without opening them, a decided safety advantage if the fill chemical is a hazardous substance like a chemical warfare agent or an explosive. The PINS Spectrum Identification Guide is intended as a reference for technical professionals responsible for the interpretation of PINS gamma-ray spectra. The guide is divided into two parts. The three chapters that constitute Part I cover the science and technology of PINS. Neutron activation analysis is the focus of Chapter 1. Chapter 2 explores PINS hardware, software, and related operational issues. Gamma-ray spectral analysis basics are introduced in Chapter 3. The six chapters of Part II cover the identification of PINS spectra in detail. Like the PINS decision tree logic, these chapters are organized by chemical element: phosphorus-based chemicals, chlorine-based chemicals, etc. These descriptions of hazardous, toxic, and/or explosive chemicals conclude with a chapter on the identification of the inert chemicals, e.g. sand, used to fill practice munitions.

Improved pinning regime by energetic ions using reduction of pinning potential

Energy Technology Data Exchange (ETDEWEB)

Weinstein, Roy; Gandini, Alberto; Sawh, Ravi-Persad; Parks, Drew; Mayes, Bill

2003-05-15

When ion damage is used to create pinning centers, full columnar pinning centers provide the largest pinning potential, U{sub pin}, but not the greatest J{sub c} or pinned field, B{sub pin}. Some of the characteristics of columnar defects which limit J{sub c} and B{sub pin} are discussed, including reduction of percolation path, and the need for a larger number of columns of damage, for pinning, than are usually estimated. It is concluded that columnar pinning centers are limited to B{sub pin}<4 T, and also severely reduce J{sub c}. Evidence is reviewed that aligned damage, or broken-columnar pinning centers, described herein, can provide orders of magnitude higher J{sub c}, and higher pinned field, despite providing lower U{sub pin}. A pinning center morphology is discussed which utilizes multiple-in-line-damage (MILD). For, e.g., present day large grain HTS J{sub c}, obtainable by MILD pinning, is estimated to be of the order of 10{sup 6} A/cm{sup 2} at 77 K, even when crystal plane alignment and weak links are not improved. Pinned field is increased by over an order of magnitude. An experiment is proposed to confirm these observations, and to directly compare MILD to columnar pinning centers. It will also determine the optimum MILD structure. Other measurements of interest, made possible by the same data set, are described.

Vph6 Mutants of Saccharomyces Cerevisiae Require Calcineurin for Growth and Are Defective in Vacuolar H(+)-Atpase Assembly

OpenAIRE

Hemenway, C. S.; Dolinski, K.; Cardenas, M. E.; Hiller, M. A.; Jones, E. W.; Heitman, J.

1995-01-01

We have characterized a Saccharomyces cerevisiae mutant strain that is hypersensitive to cyclosporin A (CsA) and FK506, immunosuppressants that inhibit calcineurin, a serine-threonine-specific phosphatase (PP2B). A single nuclear mutation, designated cev1 for calcineurin essential for viability, is responsible for the CsA-FK506-sensitive phenotype. The peptidyl-prolyl cis-trans isomerases cyclophilin A and FKBP12, respectively, mediate CsA and FK506 toxicity in the cev1 mutant strain. We demo...

Screening and selection of wild strains for L-arabinose isomerase production

Directory of Open Access Journals (Sweden)

R. M. Manzo

2013-12-01

Full Text Available The majority of L-arabinose isomerases have been isolated by recombinant techniques, but this methodology implies a reduced technological application. For this reason, 29 bacterial strains, some of them previously characterized as L-arabinose isomerase producers, were assayed as L-arabinose fermenting strains by employing conveniently designed culture media with 0.5% (w/v L-arabinose as main carbon source. From all evaluated bacterial strains, Enterococcus faecium DBFIQ ID: E36, Enterococcus faecium DBFIQ ID: ETW4 and Pediococcus acidilactici ATCC ID: 8042 were, in this order, the best L-arabinose fermenting strains. Afterwards, to assay L-arabinose metabolization and L-arabinose isomerase activity, cell-free extract and saline precipitated cell-free extract of the three bacterial cultures were obtained and the production of ketoses was determined by the cysteine carbazole sulfuric acid method. Results showed that the greater the L-arabinose metabolization ability, the higher the enzymatic activity achieved, so Enterococcus faecium DBFIQ ID: E36 was selected to continue with production, purification and characterization studies. This work thus describes a simple microbiological method for the selection of L-arabinose fermenting bacteria for the potential production of the enzyme L-arabinose isomerase.

[Influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin].

Science.gov (United States)

Shen, Congxiang; Liu, Yanhui; Wen, Zhong; Yang, Keke; Li, Guanxue; Zhang, Shenhua; Zhang, Xinyu

2015-06-23

To explore the influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin. Transfected nasopharyngeal carcinoma 5-8F cell lines with pCDH-CMV-PinX1-copGFP vector constructed by lentivirus to generate Lenti-PinX1-5-8F cells containing PinX1 gene, using Lenti-Ctrl-5-8F cell (blank vector without PinX1 gene was used to transfect 5-8F cell lines) and 5-8F cell as controls. Expression of PinX1 gene, telomerase activity, the inhibition of cancer cells proliferation, combined anticancer effect with Cisplatin and the expression of lung resistance protein (LRP) and Bcl-2 were detected with fluorescent quantitation polymerase chain reaction (PCR), flow cytometry, thiazolyl blue (MTT) method, areole test, Western blot and drug sensitivity test, respectively, in four groups (Lenti-PinX1-5-8F cell + Cisplatin, Lenti-PinX1-5-8F cell, Cisplatin and 5-8F cell) so as to explore the influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin. The telomerase activity in Lenti-PinX1-5-8F cell (0.146 ± 0.004) was lower than those in the other two control cells (Lenti-Ctrl-5-8F cell: 0.967 ± 0.016, 5-8F cell: 1.000 ± 0.034, both P Cisplatin after lower level telomerase activity induced by PinX1 gene. Proliferation index (PI) (%) in Lenti-PinX1-5-8F cell + Cisplatin (14.39 ± 3.66) was also less than the other groups (Lenti-PinX1-5-8F cell, Cisplatin and 5-8F cell groups, 32.97 ± 3.00, 31.18 ± 4.24 and 47.19 ± 4.19, all P Cisplatin, which may be mediated by the down-regulation of telomerase activity and the inhibition of LRP and Bcl-2 gene in nasopharyngeal carcinoma cells.

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.

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

The secreted L-arabinose isomerase displays anti-hyperglycemic effects in mice.

Science.gov (United States)

Rhimi, Moez; Bermudez-Humaran, Luis G; Huang, Yuan; Boudebbouze, Samira; Gaci, Nadia; Garnier, Alexandrine; Gratadoux, Jean-Jacques; Mkaouar, Héla; Langella, Philippe; Maguin, Emmanuelle

2015-12-21

The L-arabinose isomerase is an intracellular enzyme which converts L-arabinose into L-ribulose in living systems and D-galactose into D-tagatose in industrial processes and at industrial scales. D-tagatose is a natural ketohexose with potential uses in pharmaceutical and food industries. The D-galactose isomerization reaction is thermodynamically equilibrated, and leads to secondary subproducts at high pH. Therefore, an attractive L-arabinose isomerase should be thermoactive and acidotolerant with high catalytic efficiency. While many reports focused on the set out of a low cost process for the industrial production of D-tagatose, these procedures remain costly. When compared to intracellular enzymes, the production of extracellular ones constitutes an interesting strategy to increase the suitability of the biocatalysts. The L-arabinose isomerase (L-AI) from Lactobacillus sakei was expressed in Lactococcus lactis in fusion with the signal peptide of usp45 (SP(Usp45)). The L-AI protein and activity were detected only in the supernatant of the induced cultures of the recombinant L. lactis demonstrating the secretion in the medium of the intracellular L. sakei L-AI in an active form. Moreover, we showed an improvement in the enzyme secretion using either (1) L. lactis strains deficient for their two major proteases, ClpP and HtrA, or (2) an enhancer of protein secretion in L. lactis fused to the recombinant L-AI with the SP(Usp45). Th L-AI enzyme secreted by the recombinant L. lactis strains or produced intracellularly in E. coli, showed the same functional properties than the native enzyme. Furthermore, when mice are fed with the L. lactis strain secreting the L-AI and galactose, tagatose was produced in vivo and reduced the glycemia index. We report for the first time the secretion of the intracellular L-arabinose isomerase in the supernatant of food grade L. lactis cultures with hardly display other secreted proteins. The secreted L-AI originated from the food

High-speed 1.3 -1.55 um InGaAs/InP PIN photodetector for microwave photonics

Science.gov (United States)

Kozyreva, O. A.; Solov'ev, Y. V.; Polukhin, I. S.; Mikhailov, A. K.; Mikhailovskiy, G. A.; Odnoblyudov, M. A.; Gareev, E. Z.; Kolodeznyi, E. S.; Novikov, I. I.; Karachinsky, L. Ya; Egorov, A. Yu; Bougrov, V. E.

2017-11-01

We have fabricated the 1.3-1.55 um PIN photodetector based on InGaAs/InP heterostructure. Measurement results of optical and electrical characteristics of PIN photodetector chip were the following: photoconductivity at 1550 nm was 0.65 A/W and internal capacitance was 0.025 pF. Microwave model of photodetector was developed and verified by measurements of scattering matrix. The implementation of broadband (up to 20 GHz) hybrid integrated matching and biasing circuit for high-speed photodetector is presented.

Xylose isomerase improves growth and ethanol production rates from biomass sugars for both Saccharomyces pastorianus and Saccharomyces cerevisiae.

Science.gov (United States)

Miller, Kristen P; Gowtham, Yogender Kumar; Henson, J Michael; Harcum, Sarah W

2012-01-01

The demand for biofuel ethanol made from clean, renewable nonfood sources is growing. Cellulosic biomass, such as switch grass (Panicum virgatum L.), is an alternative feedstock for ethanol production; however, cellulosic feedstock hydrolysates contain high levels of xylose, which needs to be converted to ethanol to meet economic feasibility. In this study, the effects of xylose isomerase on cell growth and ethanol production from biomass sugars representative of switch grass were investigated using low cell density cultures. The lager yeast species Saccharomyces pastorianus was grown with immobilized xylose isomerase in the fermentation step to determine the impact of the glucose and xylose concentrations on the ethanol production rates. Ethanol production rates were improved due to xylose isomerase; however, the positive effect was not due solely to the conversion of xylose to xylulose. Xylose isomerase also has glucose isomerase activity, so to better understand the impact of the xylose isomerase on S. pastorianus, growth and ethanol production were examined in cultures provided fructose as the sole carbon. It was observed that growth and ethanol production rates were higher for the fructose cultures with xylose isomerase even in the absence of xylose. To determine whether the positive effects of xylose isomerase extended to other yeast species, a side-by-side comparison of S. pastorianus and Saccharomyces cerevisiae was conducted. These comparisons demonstrated that the xylose isomerase increased ethanol productivity for both the yeast species by increasing the glucose consumption rate. These results suggest that xylose isomerase can contribute to improved ethanol productivity, even without significant xylose conversion. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

21 CFR 862.1570 - Phosphohexose isomerase test system.

Science.gov (United States)

2010-04-01

.... Measurements of phosphohexose isomerase are used in the diagnosis and treatment of muscle diseases such as muscular dystrophy, liver diseases such as hepatitis or cirrhosis, and metastatic carcinoma. (b...

A physico-genetic module for the polarisation of auxin efflux carriers PIN-FORMED (PIN)

Science.gov (United States)

Hernández-Hernández, Valeria; Barrio, Rafael A.; Benítez, Mariana; Nakayama, Naomi; Romero-Arias, José Roberto; Villarreal, Carlos

2018-05-01

Intracellular polarisation of auxin efflux carriers is crucial for understanding how auxin gradients form in plants. The polarisation dynamics of auxin efflux carriers PIN-FORMED (PIN) depends on both biomechanical forces as well as chemical, molecular and genetic factors. Biomechanical forces have shown to affect the localisation of PIN transporters to the plasma membrane. We propose a physico-genetic module of PIN polarisation that integrates biomechanical, molecular, and cellular processes as well as their non-linear interactions. The module was implemented as a discrete Boolean model and then approximated to a continuous dynamic system, in order to explore the relative contribution of the factors mediating PIN polarisation at the scale of single cell. Our models recovered qualitative behaviours that have been experimentally observed and enable us to predict that, in the context of PIN polarisation, the effects of the mechanical forces can predominate over the activity of molecular factors such as the GTPase ROP6 and the ROP-INTERACTIVE CRIB MOTIF-CONTAINING PROTEIN RIC1.

The secreted l-arabinose isomerase displays anti-hyperglycemic effects in mice

OpenAIRE

Rhimi, Moez; Bermudez-Humaran, Luis G.; Huang, Yuan; Boudebbouze, Samira; Gaci, Nadia; Garnier, Alexandrine; Gratadoux, Jean-Jacques; Mkaouar, H?la; Langella, Philippe; Maguin, Emmanuelle

2015-01-01

Background The l-arabinose isomerase is an intracellular enzyme which converts l-arabinose into l-ribulose in living systems and d-galactose into d-tagatose in industrial processes and at industrial scales. d-tagatose is a natural ketohexose with potential uses in pharmaceutical and food industries. The d-galactose isomerization reaction is thermodynamically equilibrated, and leads to secondary subproducts at high pH. Therefore, an attractive l-arabinose isomerase should be thermoactive and a...

A Slow Conformational Switch in the BMAL1 Transactivation Domain Modulates Circadian Rhythms.

Science.gov (United States)

Gustafson, Chelsea L; Parsley, Nicole C; Asimgil, Hande; Lee, Hsiau-Wei; Ahlbach, Christopher; Michael, Alicia K; Xu, Haiyan; Williams, Owen L; Davis, Tara L; Liu, Andrew C; Partch, Carrie L

2017-05-18

The C-terminal transactivation domain (TAD) of BMAL1 (brain and muscle ARNT-like 1) is a regulatory hub for transcriptional coactivators and repressors that compete for binding and, consequently, contributes to period determination of the mammalian circadian clock. Here, we report the discovery of two distinct conformational states that slowly exchange within the dynamic TAD to control timing. This binary switch results from cis/trans isomerization about a highly conserved Trp-Pro imide bond in a region of the TAD that is required for normal circadian timekeeping. Both cis and trans isomers interact with transcriptional regulators, suggesting that isomerization could serve a role in assembling regulatory complexes in vivo. Toward this end, we show that locking the switch into the trans isomer leads to shortened circadian periods. Furthermore, isomerization is regulated by the cyclophilin family of peptidyl-prolyl isomerases, highlighting the potential for regulation of BMAL1 protein dynamics in period determination. Copyright © 2017 Elsevier Inc. All rights reserved.

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

Energy Technology Data Exchange (ETDEWEB)

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.

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.

Etude par génétique inverse du gène codant la protéine TARGET OF RAPAMYCIN d'Arabidopsis thaliana (AtTOR), l'homologue d'une kinase contrôlant la croissance cellulaire chez les eucaryotes

OpenAIRE

Menand, Benoit

2002-01-01

TOR (target of rapamycin) protein kinases were identified in yeast, mammals and Drosophila as central controllers of cell growth. Thu, G1 to S phases progression through the cell cycle is blocked by rapamycin, a drug which specifically inhibits TOR activity by forming a ternary complex with the peptidyl-prolyl isomerase FKBP12 (FK506 and rapamycin binding protein), and the FKBP-rapamycin binding domain (FRB) of TOR proteins. This work presents the study, the Arabidopsis homologue of yeast and...

Structure of ribose 5-phosphate isomerase from the probiotic bacterium Lactobacillus salivarius UCC118.

Science.gov (United States)

Lobley, Carina M C; Aller, Pierre; Douangamath, Alice; Reddivari, Yamini; Bumann, Mario; Bird, Louise E; Nettleship, Joanne E; Brandao-Neto, Jose; Owens, Raymond J; O'Toole, Paul W; Walsh, Martin A

2012-12-01

The structure of ribose 5-phosphate isomerase from the probiotic bacterium Lactobacillus salivarius UCC188 has been determined at 1.72 Å resolution. The structure was solved by molecular replacement, which identified the functional homodimer in the asymmetric unit. Despite only showing 57% sequence identity to its closest homologue, the structure adopted the typical α and β D-ribose 5-phosphate isomerase fold. Comparison to other related structures revealed high homology in the active site, allowing a model of the substrate-bound protein to be proposed. The determination of the structure was expedited by the use of in situ crystallization-plate screening on beamline I04-1 at Diamond Light Source to identify well diffracting protein crystals prior to routine cryocrystallography.

Design of a terahertz CW photomixer based on PIN and superlattice PIN devices

DEFF Research Database (Denmark)

Krozer, Viktor; Eichhorn, Finn

2006-01-01

We present the design of a photomixer LO based on standard and superlattice PIN diodes, operating at 1 THz. The design is based on a direct integration of a double slot antenna with the PIN device and a suitable matching circuit. The antenna has been designed together with a dielectric lens using...... Ansoft HFSS EM simulation. The large-signal PIN diode model employed in the work has been improved compared to our previously developed model presented earlier in a 3 THz design. We demonstrate that the antenna characteristic changes drastically with the device in place....

Heterogeneous neutron-leakage model for PWR pin-by-pin calculation

International Nuclear Information System (INIS)

Li, Yunzhao; Zhang, Bin; Wu, Hongchun; Shen, Wei

2017-01-01

Highlights: •The derivation of the formula of the leakage model is introduced. This paper evaluates homogeneous and heterogeneous leakage models used in PWR pin-by-pin calculation. •The implements of homogeneous and heterogeneous leakage models used in pin-cell homogenization of the lattice calculation are studied. A consistent method of cooperation between the heterogeneous leakage model and the pin-cell homogenization theory is proposed. •Considering the computational cost, a new buckling search scheme is proposed to reach the convergence faster. The computational cost of the newly proposed neutron balance scheme is much less than the power-method scheme and the linear-interpolation scheme. -- Abstract: When assembly calculation is performed with the reflective boundary condition, a leakage model is usually required in the lattice code. The previous studies show that the homogeneous leakage model works effectively for the assembly homogenization. However, it becomes different and unsettled for the pin-cell homogenization. Thus, this paper evaluates homogeneous and heterogeneous leakage models used in pin-by-pin calculation. The implements of homogeneous and heterogeneous leakage models used in pin-cell homogenization of the lattice calculation are studied. A consistent method of cooperation between the heterogeneous leakage model and the pin-cell homogenization theory is proposed. Considering the computational cost, a new buckling search scheme is proposed to reach the convergence faster. For practical reactor-core applications, the diffusion coefficients determined by the transport cross-section or by the leakage model are compared with each other to determine which one is more accurate for the Pressurized Water Reactor pin-by-pin calculation. Numerical results have demonstrated that the heterogeneous leakage model together with the diffusion coefficient determined by the heterogeneous leakage model would have the higher accuracy. The new buckling search

Glucose(xylose isomerase production by Streptomyces sp. CH7 grown on agricultural residues

Directory of Open Access Journals (Sweden)

Kankiya Chanitnun

2012-09-01

Full Text Available Streptomyces sp. CH7 was found to efficiently produce glucose(xylose isomerase when grown on either xylan or agricultural residues. This strain produced a glucose(xylose isomerase activity of roughly 1.8 U/mg of protein when it was grown in medium containing 1% xylose as a carbon source. Maximal enzymatic activities of about 5 and 3 U/mg were obtained when 1% xylan and 2.5% corn husks were used, respectively. The enzyme was purified from a mycelial extract to 16-fold purity with only two consecutive column chromatography steps using Macro-prep DEAE and Sephacryl-300, respectively. The approximate molecular weight of the purified enzyme is 170 kDa, and it has four identical subunits of 43.6 kDa as estimated by SDS-PAGE. Its Km values for glucose and xylose were found to be 258.96 and 82.77 mM, respectively, and its Vmax values are 32.42 and 63.64 μM/min/mg, respectively. The purified enzyme is optimally active at 85ºC and pH 7.0. It is stable at pH 5.5-8.5 and at temperatures up to 60ºC after 30 min. These findings indicate that glucose(xylose isomerase from Streptomyces sp. CH7 has the potential for industrial applications, especially for high-fructose syrup production and bioethanol fermentation from hemicellulosic hydrolysates by Saccharomyces cerevisiae.

Unicortical self-drilling external fixator pins reduce thermal effects during pin insertion.

Science.gov (United States)

Greinwald, Markus; Varady, Patrick A; Augat, Peter

2017-12-14

External fixation is associated with the risk of pin loosening and pin infection potentially associated to thermal bone necrosis during pin insertion. This study aims to investigate if the use of external fixator systems with unicortical pins reduces the heat production during pin insertion compared to fixators with bicortical pins. Porcine bone specimens were employed to determine bone temperatures during insertion of fixator pins. Two thermographic cameras were used for a simultaneous temperature measurement on the bone surface (top view) and a bone cross-section (front view). Self-drilling unicortical and bicortical pins were inserted at different rotational speeds: (30-600) rpm. Maximum and mean temperatures of the emerging bone debris, bone surface and bone cross-section were analyzed. Maximum temperatures of up to 77 ± 26 °C were measured during pin insertion in the emerging debris and up to 42 ± 2 °C on the bone surface. Temperatures of the emerging debris increased with increasing rotational speeds. Bicortical pin insertion generated significantly higher temperatures at low insertion speed (30 rpm) CONCLUSION: The insertion of external fixator pins can generate a considerable amount of heat around the pins, primarily emerging from bone debris and at higher insertion speeds. Our findings suggest that unicortical, self-drilling fixator pins have a decreased risk for thermal damage, both to the surrounding tissue and to the bone itself.

Purification, crystallization and preliminary crytallographic analysis of phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus

NARCIS (Netherlands)

Akerboom, A.P.; Turnbull, A.P.; Hargreaves, D.; Fischer, M.; Geus, de D.; Sedelnikova, S.E.; Berrisford, J.M.; Baker, P.J.; Verhees, C.H.; Oost, van der J.; Rice, D.W.

2003-01-01

The glycolytic enzyme phosphoglucose isomerase catalyses the reversible isomerization of glucose 6-phosphate to fructose 6-phosphate. The phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus, which shows no sequence similarity to any known bacterial or eukaryotic

Preparation of fluorescence quenched libraries containing interchain disulphide bonds for studies of protein disulphide isomerases

DEFF Research Database (Denmark)

Spetzler, J C; Westphal, V; Winther, Jakob R.

1998-01-01

Protein disulphide isomerase is an enzyme that catalyses disulphide redox reactions in proteins. In this paper, fluorogenic and interchain disulphide bond containing peptide libraries and suitable substrates, useful in the study of protein disulphide isomerase, are described. In order to establish...... the quenching chromophore (Tyr(NO2)) and Cys(pNpys) activated for reaction with a second thiol. The formation and cleavage of the interchain disulphide bonds in the library were monitored under a fluorescence microscope. Substrates to investigate the properties of protein disulphide isomerase in solution were...

Studies on the production of glucose isomerase by Bacillus licheniformis

Directory of Open Access Journals (Sweden)

Nwokoro Ogbonnaya

2015-09-01

Full Text Available This work reports the effects of some culture conditions on the production of glucose isomerase by Bacillus licheniformis. The bacterium was selected based on the release of 3.62 mg/mL fructose from the fermentation of glucose. Enzyme was produced using a variety of carbon substrates but the highest enzyme activity was detected in a medium containing 0.5% xylose and 1% glycerol (specific activity = 6.88 U/mg protein. Media containing only xylose or glucose gave lower enzyme productivies (specific activities= 4.60 and 2.35 U/mg protein respectively. The effects of nitrogen substrates on glucose isomerase production showed that yeast extract supported maximum enzyme activity (specific activity = 5.24 U/mg protein. Lowest enzyme activity was observed with sodium trioxonitrate (specific activity = 2.44 U/mg protein. In general, organic nitrogen substrates supported higher enzyme productivity than inorganic nitrogen substrates. Best enzyme activity was observed in the presence of Mg2+ (specific activity = 6.85 U/mg protein while Hg2+ was inhibitory (specific activity = 1.02 U/mg protein. The optimum pH for best enzyme activity was 6.0 while optimum temperature for enzyme production was 50ºC.

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

Affinity labeling and characterization of the active site histidine of glucosephosphate isomerase

International Nuclear Information System (INIS)

Gibson, D.R.; Gracy, R.W.; Hartman, F.C.

1980-01-01

N-bromoacetylethanolamine phosphate was found to act as a specific affinity label for the active center of glucosephosphate isomerase. The inactivation process followed pseudo-first order kinetics, was irreversible, and exhibited rate saturation kinetics with minimal half-lives of inactivation of 4.5 and 6.3 min for the enzyme isolated from human placenta and rabbit muscle, respectively. The pH dependence of the inactivation process closely paralleled the pH dependence of the overall catalytic process with pK/sub a/ values at pH 6.4 and 9.0. The stoichiometry of labeling of either enzyme, as determined with N-bromo[ 14 C 2 ]acetylethanolamine phosphate, was 1 eq of the affinity label/subunit of enzyme. After acid hydrolysis and amino acid analysis of the radioactive affinity-labeled human enzyme, only radioactive 3-carboxymethyl histidine was found. In the case of the rabbit enzyme, the only radioactive derivative obtained was 1-carboxymethyl histidine. Active site tryptic peptides were isolated by solvent extraction, thin layer peptide fingerprinting, and ion exchange chromatography before and after removal of the phosphate from the active site peptide. Amino acid analysis of the labeled peptides from the two species were very similar. Using high sensitivity methods for sequence analysis, the primary structure of the active site was established as Val-Leu-His-Ala-Glu-Asn-Val-Asp (Gly,Thr,Ser) Glu-Ile (Thr-Gly-His-Lys-Glx)-Tyr-Phe. Apparent sequence homology between the catalytic center of glucosephosphate isomerase and triosephosphate isomerase suggest that the two enzymes may have evolved from a common ancestral gene

Regulation of AR and (beta)-Catenin Signaling by Pin 1 in Prostate Cancer

National Research Council Canada - National Science Library

Chen, Shaoyong

2006-01-01

.... The mechanisms include that Pin1 can enhance beta-catenin nuclear localization, TCF/beta-catenin dependent Topflash activity, and c-Myc and Cyclin D1 expression, and disrupt AR-mediated suppression...

Equilibrium vortex structures of type-II/1 superconducting films with washboard pinning landscapes

Science.gov (United States)

Wei, C. A.; Xu, X. B.; Xu, X. N.; Wang, Z. H.; Gu, M.

2018-05-01

We numerically study the equilibrium vortex structures of type-II/1 superconducting films with a periodic quasi-one-dimensional corrugated substrate. We show as a function of substrate period and pinning strength that, the vortex system displays a variety of vortex phases including arrays consisted of vortex clumps with different morphologies, ordered vortex stripes parallel and perpendicular to pinning troughs, and ordered one-dimensional vortex chains. Our simulations are helpful in understanding the structural modulations for extensive systems with both competing interactions and competing periodicities.

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).

Inhibition of histone deacetylation alters Arabidopsis root growth in response to auxin via PIN1 degradation.

Science.gov (United States)

Nguyen, Hoai Nguyen; Kim, Jun Hyeok; Jeong, Chan Young; Hong, Suk-Whan; Lee, Hojoung

2013-10-01

Our results showed the histone deacetylase inhibitors (HDIs) control root development in Arabidopsis via regulation of PIN1 degradation. Epigenetic regulation plays a crucial role in the expression of many genes in response to exogenous or endogenous signals in plants as well as other organisms. One of epigenetic mechanisms is modifications of histone, such as acetylation and deacetylation, are catalyzed by histone acetyltransferase (HAT) and histone deacetylase (HDAC), respectively. The Arabidopsis HDACs, HDA6, and HDA19, were reported to function in physiological processes, including embryo development, abiotic stress response, and flowering. In this study, we demonstrated that histone deacetylase inhibitors (HDIs) inhibit primary root elongation and lateral root emergence. In response to HDIs treatment, the PIN1 protein was almost abolished in the root tip. However, the PIN1 gene did not show decreased expression in the presence of HDIs, whereas IAA genes exhibited increases in transcript levels. In contrast, we observed a stable level of gene expression of stress markers (KIN1 and COR15A) and a cell division marker (CYCB1). Taken together, these results suggest that epigenetic regulation may control auxin-mediated root development through the 26S proteasome-mediated degradation of PIN1 protein.

Magnetic pinning in superconductor-ferromagnet multilayers

International Nuclear Information System (INIS)

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

2000-01-01

We argue that superconductor/ferromagnet multilayers of nanoscale period should exhibit strong pinning of vortices by the magnetic domain structure in magnetic fields below the coercive field when ferromagnetic layers exhibit strong perpendicular magnetic anisotropy. The estimated maximum magnetic pinning energy for single vortex in such a system is about 100 times larger than the pinning energy by columnar defects. This pinning energy may provide critical currents as high as 10 6 -10 7 A/cm 2 at high temperatures (but not very close to T c ) at least in magnetic fields below 0.1 T. (c) 2000 American Institute of Physics

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

Fabrication of FFTF fuel pin wire wrap

International Nuclear Information System (INIS)

Epperson, E.M.

1980-06-01

Lateral spacing between FFTF fuel pins is required to provide a passageway for the sodium coolant to flow over each pin to remove heat generated by the fission process. This spacing is provided by wrapping each fuel pin with type 316 stainless steel wire. This wire has a 1.435mm (0.0565 in.) to 1.448mm (0.0570 in.) diameter, contains 17 +- 2% cold work and was fabricated and tested to exacting RDT Standards. About 500 kg (1100 lbs) or 39 Km (24 miles) of fuel pin wrap wire is used in each core loading. Fabrication procedures and quality assurance tests are described

Bacterial L-arabinose isomerases: industrial application for D-tagatose production.

Science.gov (United States)

Boudebbouze, Samira; Maguin, Emmanuelle; Rhimi, Moez

2011-12-01

D-tagatose is a natural monosaccharide with a low caloric value and has an anti-hyperglycemiant effect. This hexose has potential applications both in pharmaceutical and agro-food industries. However, the use of D-tagatose remains limited by its production cost. Many production procedures including chemical and biological processes were developed and patented. The most profitable production way is based on the use of L-arabinose isomerase which allows the manufacture of D-tagatose with an attractive rate. Future developments are focused on the generation of L-arabinose isomerases having biochemical properties satisfying the industrial applications. This report provides a brief review of the most recent patents that have been published relating to this area.

Multicenter Study of Pin Site Infections and Skin Complications Following Pinning of Pediatric Supracondylar Humerus Fractures.

Science.gov (United States)

Combs, Kristen; Frick, Steven; Kiebzak, Gary

2016-12-03

Pediatric supracondylar humerus fractures are the most common elbow fractures in pediatric patients. Surgical fixation using pins is the primary treatment for displaced fractures. Pin site infections may follow supracondylar humerus fracture fixation; the previously reported incidence rate in the literature is 2.34%, but there is significant variability in reported incidence rates of pin site infection. This study aims to define the incidence rate and determine pre-, peri-, and postoperative factors that may contribute to pin site infection following operative reduction, pinning, and casting. A retrospective chart analysis was performed over a one-year period on patients that developed pin site infection. A cast care form was added to Nemours' electronic medical records (EMR) system (Epic Systems Corp., Verona, WI) to identify pin site infections for retrospective review. The cast care form noted any inflamed or infected pins. Patients with inflamed or infected pin sites underwent a detailed chart review. Preoperative antibiotic use, number and size of pins used, method of postoperative immobilization, pin dressings, whether postoperative immobilization was changed prior to pin removal, and length of time pins were in place was recorded. A total of 369 patients underwent operative reduction, pinning, and casting. Three patients developed a pin site infection. The pin site infection incidence rate was 3/369=0.81%. Descriptive statistics were reported for the three patients that developed pin site infections and three patients that developed pin site complications. Pin site infection development is low. Factors that may contribute to the development of pin site infection include preoperative antibiotic use, length of time pins are left in, and changing the cast prior to pin removal.

Magnetic pinning in superconductor-ferromagnet multilayers

Energy Technology Data Exchange (ETDEWEB)

Bulaevskii, L. N. [Department of Physics and Astronomy, CUNY Lehman College 250 Bedford Park Boulevard West, Bronx, New York 10468-1589 (United States); Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States); Chudnovsky, E. M. [Department of Physics and Astronomy, CUNY Lehman College, 250 Bedford Park Boulevard West, Bronx, New York 10468-1589 (United States); Maley, M. P. [Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States)

2000-05-01

We argue that superconductor/ferromagnet multilayers of nanoscale period should exhibit strong pinning of vortices by the magnetic domain structure in magnetic fields below the coercive field when ferromagnetic layers exhibit strong perpendicular magnetic anisotropy. The estimated maximum magnetic pinning energy for single vortex in such a system is about 100 times larger than the pinning energy by columnar defects. This pinning energy may provide critical currents as high as 10{sup 6}-10{sup 7} A/cm{sup 2} at high temperatures (but not very close to T{sub c}) at least in magnetic fields below 0.1 T. (c) 2000 American Institute of Physics.

A macroscopic cross-section model for BWR pin-by-pin core analysis

International Nuclear Information System (INIS)

Fujita, Tatsuya; Endo, Tomohiro; Yamamoto, Akio

2014-01-01

A macroscopic cross-section model used in boiling water reactor (BWR) pin-by-pin core analysis is studied. In the pin-by-pin core calculation method, pin-cell averaged cross sections are calculated for many combinations of core state and depletion history variables and are tabulated prior to core calculations. Variations of cross sections in a core simulator are caused by two different phenomena (i.e. instantaneous and history effects). We treat them through the core state variables and the exposure-averaged core state variables, respectively. Furthermore, the cross-term effect among the core state and the depletion history variables is considered. In order to confirm the calculation accuracy and discuss the treatment of the cross-term effect, the k-infinity and the pin-by-pin fission rate distributions in a single fuel assembly geometry are compared. Some cross-term effects could be negligible since the impacts of them are sufficiently small. However, the cross-term effects among the control rod history (or the void history) and other variables have large impacts; thus, the consideration of them is crucial. The present macroscopic cross-section model, which considers such dominant cross-term effects, well reproduces the reference results and can be a candidate in practical applications for BWR pin-by-pin core analysis on the normal operations. (author)

Methods of measuring Protein Disulfide Isomerase activity: a critical overview

Science.gov (United States)

Watanabe, Monica; Laurindo, Francisco; Fernandes, Denise

2014-09-01

Protein disulfide isomerase is an essential redox chaperone from the endoplasmic reticulum (ER) and is responsible for correct disulfide bond formation in nascent proteins. PDI is also found in other cellular locations in the cell, particularly the cell surface. Overall, PDI contributes to ER and global cell redox homeostasis and signaling. The knowledge about PDI structure and function progressed substantially based on in vitro studies using recombinant PDI and chimeric proteins. In these experimental scenarios, PDI reductase and chaperone activities are readily approachable. In contrast, assays to measure PDI isomerase activity, the hallmark of PDI family, are more complex. Assessment of PDI roles in cells and tissues mainly relies on gain- or loss-of-function studies. However, there is limited information regarding correlation of experimental readouts with the distinct types of PDI activities. In this mini-review, we evaluate the main methods described for measuring the different kinds of PDI activity: thiol reductase, thiol oxidase, thiol isomerase and chaperone. We emphasize the need to use appropriate controls and the role of critical interferents (e.g., detergent, presence of reducing agents). We also discuss the translation of results from in vitro studies with purified recombinant PDI to cellular and tissue samples, with critical comments on the interpretation of results.

Effect of pH on simultaneous saccharification and isomerization by glucoamylase and glucose isomerase.

Science.gov (United States)

Mishra, Abha; Debnath Das, Meera

2002-01-01

pH and temperature play critical roles in multistep enzymatic conversions. In such conversions, the optimal pH for individual steps differs greatly. In this article, we describe the production of glucoamylase (from Aspergillus oryzae MTCC152 in solid-state fermentation) and glucose isomerase (from Streptomyces griseus NCIM2020 in submerged fermentation), used in industries for producing high-fructose syrup. Optimum pH for glucoamylase was found to be 5.0. For glucose isomerase, the optimum pH ranged between 7.0 and 8.5, depending on the type of buffer used. Optimum temperature for glucoamylase and glucose isomerase was 50 and 60 degrees C, respectively. When both the enzymatic conversions were performed simultaneously at a compromised pH of 6.5, both the enzymes showed lowered activity. We also studied the kinetics at different pHs, which allows the two-step reaction to take place simultaneously. This was done by separating two steps by a thin layer of urease. Ammonia generated by the hydrolysis of urea consumed the hydrogen ions, thereby allowing optimal activity of glucose isomerase at an acidic pH of 5.0.

Critical experiments supporting underwater storage of tightly packed configurations of spent fuel pins. Technical progress report, January 1-March 31, 1981

International Nuclear Information System (INIS)

Hoovler, G.S.; Baldwin, M.N.

1981-04-01

Critical experiments are in progress on arrays of 2 1/2% enriched UO 2 fuel pins simulating underwater pin storage of spent power reactor fuel. Pin storage refers to a spent fuel storage concept in which the fuel assemblies are dismantled and the fuel pins are tightly packed into specially designed canisters. These experiments are providing benchmark data with which to validate nuclear codes used to design spent fuel pin storage racks

Characteristics of chalcone isomerase promoter in crabapple leaves ...

African Journals Online (AJOL)

Anthocyanins are secondary metabolites found in higher plants that contribute to the colors of plants and chalcone isomerase (CHI) is one of the key enzymes in anthocyanin biosynthetic pathway. What characteristic is CHI promoter known as the regulation sequence of CHI gene, has been rarely investigated. We isolated A ...

Biochemical properties of L-arabinose isomerase from Clostridium hylemonae to produce D-tagatose as a functional sweetener.

Science.gov (United States)

Nguyen, Tien-Kieu; Hong, Moon-Gi; Chang, Pahn-Shick; Lee, Byung-Hoo; Yoo, Sang-Ho

2018-01-01

d-Tagatose has gained substantial interest due to its potential functionalities as a sucrose substitute. In this study, the gene araA, encoding l-arabinose isomerase (l-AI) from Clostridium hylemonae (DSM 15053), was cloned and expressed in Escherichia coli BL21 (DE3). This gene consists of 1,506 nucleotides and encodes a protein of 501 amino acid residues with a calculated molecular mass of 56,554 Da. Since l-AI was expressed as an intracellular inclusion body, this enzyme was solubilized with guanidine hydrochloride, refolded, and activated with a descending concentration gradient of urea. The purified enzyme exhibited the greatest activity at 50°C, pH 7-7.5, and required 1 mM of Mg2+ as a cofactor. Notably, the catalytic efficiency (3.69 mM-1sec-1) of l-AI from C. hylemonae on galactose was significantly greater than that of other previously reported enzymes. The bioconversion yield of d-tagatose using the C. hylemonae l-arabinose isomerase at 60°C reached approximately 46% from 10 mM of d-galactose after 2 h. From these results, it is suggested that the l-arabinose isomerase from C. hylemonae could be utilized as a potential enzyme for d-tagatose production due to its high conversion yield at an industrially competitive temperature.

Transformation of Helicopter PinS Procedures for Airplanes

Directory of Open Access Journals (Sweden)

Jakub Kraus

2013-09-01

Full Text Available This article deals with the possibility to use existing helicopter Point in Space procedures with minor changes for airplanes. The basis is to find parts of PinS procedures that need to be changed, suggest these changes, and then determine whether the revised procedures could be usable and could bring the benefits for airplane operations.

Bioconversion of D-galactose into D-tagatose by expression of L-arabinose isomerase.

Science.gov (United States)

Roh, H J; Kim, P; Park, Y C; Choi, J H

2000-02-01

D-Tagatose is a potential bulking agent in food as a non-calorific sweetener. To produce D-tagatose from cheaper resources, plasmids harbouring the L-arabinose isomerase gene (araA) from Escherichia coli, Bacillus subtilis and Salmonella typhimurium were constructed because L-arabinose isomerase was suggested previously as an enzyme that mediates the bioconversion of galactose into tagatose as well as that of arabinose to ribulose. The constructed plasmids were named pTC101, pTC105 and pTC106, containing araA from E. coli, B. subtilis and S. typhimurium respectively. In the cultures of recombinant E. coli with pTC101, pTC105 and pTC106, tagatose was produced from galactose in 9.9, 7.1 and 6.9% yields respectively. The enzyme extract of E. coli with the plasmid pTC101 also converted galactose into tagatose with a 96.4% yield.

The pin pixel detector--X-ray imaging

CERN Document Server

Bateman, J E; Derbyshire, G E; Duxbury, D M; Marsh, A S; Simmons, J E; Stephenson, R

2002-01-01

The development and testing of a soft X-ray gas pixel detector, which uses connector pins for the anodes is reported. Based on a commercial 100 pin connector block, a prototype detector of aperture 25.4 mm centre dot 25.4 mm can be economically fabricated. The individual pin anodes all show the expected characteristics of small gas detectors capable of counting rates reaching 1 MHz per pin. A 2-dimensional resistive divide readout system has been developed to permit the imaging properties of the detector to be explored in advance of true pixel readout electronics.

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.

Pin clad strains in Phenix

International Nuclear Information System (INIS)

Languille, A.

1979-07-01

The Phenix reactor has operated for 4 years in a satisfactory manner. The first 2 sub-assembly loadings contained pins clad in solution treated 316. The principal pin strains are: diametral strain (swelling and irradiation creep), ovality and spiral bending of the pin (interaction of wire and pin cluster and wrapper). A pin cluster irradiated to a dose of 80 dpa F reached a pin diameter strain of 5%. This strain is principally due to swelling (low fission gas pressure). The principal parameters governing the swelling are instantaneous dose, time and temperature for a given type of pin cladding. Other types of steel are or will be irradiated in Phenix. In particular, cold-worked titanium stabilised 316 steel should contribute towards a reduction in the pin clad strains and increase the target burn-up in this reactor. (author)

Cesium chemistry in GCFR fuel pins

International Nuclear Information System (INIS)

Fee, D.C.; Johnson, C.E.

1979-01-01

The fuel rod design for the Gas Cooled Fast-Breeder Reactor (GCFR) is similar to that employed for the Liquid Metal Fast Breeder Reactor (LMFBR) with the exception of the unique features inherent to the use of helium as the coolant. These unique design features include the use of (1) vented and pressure-equalized fuel rods, and (2) ribbed cladding along 75% of the fuel section. The former design feature enables reduction in cladding thickness and prevention of possible creep collapse of the cladding due to the high coolant pressure (8.5 MPa). The latter design feature brings about improved heat transfer characteristics. Each GCFR fuel rod is vented to a manifold whereby gaseous fission products diffusing out of the fuel pin are retained on charcoal traps. As a result, the internal pressure of a GCFR fuel pin does not increase during irradiation. In addition, the venting system also maintains the pressure within the fuel pin slightly below (0.3 to 0.5 MPa) the coolant pressure outside the fuel pin. Consequently, should a breach occur in the cladding, helium flows into the breached fuel pin thereby minimizing fission product contamination of the coolant. These desirable aspects of a GCFR fuel pin can be maintained only as long as axial gas transport paths are available and operating within the fuel pin

Vortex pinning landscape in MOD-TFA YBCO nanostroctured films

Science.gov (United States)

Gutierrez, J.; Puig, T.; Pomar, A.; Obradors, X.

2008-03-01

A methodology of general validity to study vortex pinning in YBCO based on Jc transport measurements is described. It permits to identify, separate and quantify three basic vortex pinning contributions associated to anisotropic-strong, isotropic-strong and isotropic-weak pinning centers. Thereof, the corresponding vortex pinning phase diagrams are built up. This methodology is applied to the new solution-derived YBCO nanostructured films, including controlled interfacial pinning by the growth of nanostructured templates by means of self-assembled processes [1] and YBCO-BaZrO3 nanocomposites prepared by modified solution precursors. The application of the methodology and comparison with a standard solution-derived YBCO film [2], enables us to identify the nature and the effect of the additional pinning centers induced. The nanostructured templates films show c-axis pinning strongly increased, controlling most of the pinning phase diagram. On the other hand, the nanocomposites have achieved so far, the highest pinning properties in HTc-superconductors [3], being the isotropic-strong defects contribution the origin of their unique properties. [1] M. Gibert et al, Adv. Mat. vol 19, p. 3937 (2007) [2] Puig.T et al, SuST EUCAS 2007 (to be published) [3] J. Gutierrez et al, Nat. Mat. vol. 6, p. 367 (2007) * Work supported by HIPERCHEM, NANOARTIS and MAT2005-02047

Pin care

Science.gov (United States)

... Drugs & Supplements Videos & Tools Español You Are Here: Home → Medical Encyclopedia → Pin care URL of this page: //medlineplus.gov/ency/patientinstructions/000481.htm Pin care To use the sharing features on this page, please enable JavaScript. Broken bones can be fixed in surgery with metal ...

Isoform-specific inhibition of cyclophilins.

Science.gov (United States)

Daum, Sebastian; Schumann, Michael; Mathea, Sebastian; Aumüller, Tobias; Balsley, Molly A; Constant, Stephanie L; de Lacroix, Boris Féaux; Kruska, Fabian; Braun, Manfred; Schiene-Fischer, Cordelia

2009-07-07

Cyclophilins belong to the enzyme class of peptidyl prolyl cis-trans isomerases which catalyze the cis-trans isomerization of prolyl bonds in peptides and proteins in different folding states. Cyclophilins have been shown to be involved in a multitude of cellular functions like cell growth, proliferation, and motility. Among the 20 human cyclophilin isoenzymes, the two most abundant members of the cyclophilin family, CypA and CypB, exhibit specific cellular functions in several inflammatory diseases, cancer development, and HCV replication. A small-molecule inhibitor on the basis of aryl 1-indanylketones has now been shown to discriminate between CypA and CypB in vitro. CypA binding of this inhibitor has been characterized by fluorescence anisotropy- and isothermal titration calorimetry-based cyclosporin competition assays. Inhibition of CypA- but not CypB-mediated chemotaxis of mouse CD4(+) T cells by the inhibitor provided biological proof of discrimination in vivo.

Kinase-dead ATM protein is highly oncogenic and can be preferentially targeted by Topo-isomerase I inhibitors.

Science.gov (United States)

Yamamoto, Kenta; Wang, Jiguang; Sprinzen, Lisa; Xu, Jun; Haddock, Christopher J; Li, Chen; Lee, Brian J; Loredan, Denis G; Jiang, Wenxia; Vindigni, Alessandro; Wang, Dong; Rabadan, Raul; Zha, Shan

2016-06-15

Missense mutations in ATM kinase, a master regulator of DNA damage responses, are found in many cancers, but their impact on ATM function and implications for cancer therapy are largely unknown. Here we report that 72% of cancer-associated ATM mutations are missense mutations that are enriched around the kinase domain. Expression of kinase-dead ATM (Atm(KD/-)) is more oncogenic than loss of ATM (Atm(-/-)) in mouse models, leading to earlier and more frequent lymphomas with Pten deletions. Kinase-dead ATM protein (Atm-KD), but not loss of ATM (Atm-null), prevents replication-dependent removal of Topo-isomerase I-DNA adducts at the step of strand cleavage, leading to severe genomic instability and hypersensitivity to Topo-isomerase I inhibitors. Correspondingly, Topo-isomerase I inhibitors effectively and preferentially eliminate Atm(KD/-), but not Atm-proficientor Atm(-/-) leukemia in animal models. These findings identify ATM kinase-domain missense mutations as a potent oncogenic event and a biomarker for Topo-isomerase I inhibitor based therapy.

Genome-wide identification and evolution of the PIN-FORMED (PIN) gene family in Glycine max.

Science.gov (United States)

Liu, Yuan; Wei, Haichao

2017-07-01

Soybean (Glycine max) is one of the most important crop plants. Wild and cultivated soybean varieties have significant differences worth further investigation, such as plant morphology, seed size, and seed coat development; these characters may be related to auxin biology. The PIN gene family encodes essential transport proteins in cell-to-cell auxin transport, but little research on soybean PIN genes (GmPIN genes) has been done, especially with respect to the evolution and differences between wild and cultivated soybean. In this study, we retrieved 23 GmPIN genes from the latest updated G. max genome database; six GmPIN protein sequences were changed compared with the previous database. Based on the Plant Genome Duplication Database, 18 GmPIN genes have been involved in segment duplication. Three pairs of GmPIN genes arose after the second soybean genome duplication, and six occurred after the first genome duplication. The duplicated GmPIN genes retained similar expression patterns. All the duplicated GmPIN genes experienced purifying selection (K a /K s genome sequence of 17 wild and 14 cultivated soybean varieties. Our research provides useful and comprehensive basic information for understanding GmPIN genes.

Cloning and characterization of peptidylprolyl isomerase B in the ...

African Journals Online (AJOL)

Peptidylprolyl isomerases (PPIases) play essential roles in protein folding and are implicated in immune response and cell cycle control. Our previous proteomic analysis indicated that Bombyx mori PPIases may be involved in anti- Bombyx mori nucleopolyhedrovirus (BmNPV) response. To help investigate this mechanism, ...

Neutron radiography of fuel pins

International Nuclear Information System (INIS)

Jackson, C.N. Jr.; Powers, H.G.; Burgess, C.A.

1975-01-01

Neutron radiography performed with a reactor source has been shown to be a superior radiographic method for the examination of unirradiated mixed oxide fuel pins at the Hanford Engineering Development Laboratory. Approximately 1,700 fuel pins were contained in a sample that demonstrated the capability of the method for detecting laminations, structural flaws, fissile density variation, hydrogenous inclusions and voids in assembled fuel pins. The nature, extent, and importance of the detected conditions are substantiated by gamma autoradiography and by destructive analysis employing alpha autoradiography, electron microprobe and visual inspection. Also, a series of radiographs illustrate the response of neutron radiography as compared to low voltage and high voltage x-ray and gamma source Iridium 192 radiography. (U.S.)

Assessment of pin-by-pin fission rate distribution within MOX/UO{sub 2} fuel assembly using MCNPX code

Energy Technology Data Exchange (ETDEWEB)

Louis, Heba Kareem; Amin, Esmat [Nuclear and Radiological Regulation Authority (NRRA), Cairo (Egypt). Safety Engineering Dept.

2016-03-15

The aim of the present paper is to assess the calculations of pin-by-pin group integrated fission rates within MOX/UO{sub 2} Fuel assemblies using the Monte Carlo code MCNP2.7c with two sets of the available latest nuclear data libraries used for calculating MOX-fueled systems. The data that are used in this paper are based on the benchmark by the NEA Nuclear Science Committee (NSC). The k{sub ∞} and absorption/fission reaction rates per isotope, k{sub eff} and pin-by-pin group integrated fission rates on 1/8 fraction of the geometry are determined. To assess the overall pin-by-pin fission rate distribution, the collective per cent error measures were investigated. The results of AVG, MRE and RMS error measures were less than 1 % error. The present results are compared with other participants using other Monte Carlo codes and with CEA results that were taken in the benchmark as reference. The results with ENDF/B-VI.6 are close to the results received by MVP (JENDL3.2) and SCALE 4.2 (JEF2.2). The results with ENDF/BVII.1 give higher values of k{sub ∞} reflecting the changes in the newer evaluations. In almost all results presented here, the MCNP calculated results with ENDF/B VII.1 should be considered more than those obtained by using other Monte Carlo codes and nuclear data libraries. The present calculations may be consider a reference for evaluating the numerical schemes in production code systems, as well as the global performance including cross-section data reduction methods as the calculations used continuous energy and no geometrical approximations.

MTH1745, a protein disulfide isomerase-like protein from thermophilic archaea, Methanothermobacter thermoautotrophicum involving in stress response.

Science.gov (United States)

Ding, Xia; Lv, Zhen-Mei; Zhao, Yang; Min, Hang; Yang, Wei-Jun

2008-01-01

MTH1745 is a putative protein disulfide isomerase characterized with 151 amino acid residues and a CPAC active-site from the anaerobic archaea Methanothermobacter thermoautotrophicum. The potential functions of MTH1745 are not clear. In the present study, we show a crucial role of MTH1745 in protecting cells against stress which may be related to its functions as a disulfide isomerase and its chaperone properties. Using real-time polymerase chain reaction analyses, the level of MTH1745 messenger RNA (mRNA) in the thermophilic archaea M. thermoautotrophicum was found to be stress-induced in that it was significantly higher under low (50 degrees C) and high (70 degrees C) growth temperatures than under the optimal growth temperature for the organism (65 degrees C). Additionally, the expression of MTH1745 mRNA was up-regulated by cold shock (4 degrees C). Furthermore, the survival of MTH1745 expressing Escherichia coli cells was markedly higher than that of control cells in response to heat shock (51.0 degrees C). These results indicated that MTH1745 plays an important role in the resistance of stress. By assay of enzyme activities in vitro, MTH1745 also exhibited a chaperone function by promoting the functional folding of citrate synthase after thermodenaturation. On the other hand, MTH1745 was also shown to function as a disulfide isomerase on the refolding of denatured and reduced ribonuclease A. On the basis of its single thioredoxin domain, function as a disulfide isomerase, and its chaperone activity, we suggest that MTH1745 may be an ancient protein disulfide isomerase. These studies may provide clues to the understanding of the function of protein disulfide isomerase in archaea.

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.

Flux Pinning in Superconductors

CERN Document Server

Matsushita, Teruo

2007-01-01

The book covers the flux pinning mechanisms and properties and the electromagnetic phenomena caused by the flux pinning common for metallic, high-Tc and MgB2 superconductors. The condensation energy interaction known for normal precipitates or grain boundaries and the kinetic energy interaction proposed for artificial Nb pins in Nb-Ti, etc., are introduced for the pinning mechanism. Summation theories to derive the critical current density are discussed in detail. Irreversible magnetization and AC loss caused by the flux pinning are also discussed. The loss originally stems from the ohmic dissipation of normal electrons in the normal core driven by the electric field induced by the flux motion. The readers will learn why the resultant loss is of hysteresis type in spite of such mechanism. The influence of the flux pinning on the vortex phase diagram in high Tc superconductors is discussed, and the dependencies of the irreversibility field are also described on other quantities such as anisotropy of supercondu...

PIN architecture for ultrasensitive organic thin film photoconductors.

Science.gov (United States)

Jin, Zhiwen; Wang, Jizheng

2014-06-17

Organic thin film photoconductors (OTFPs) are expected to have wide applications in the field of optical communications, artificial vision and biomedical sensing due to their great advantages of high flexibility and low-cost large-area fabrication. However, their performances are not satisfactory at present: the value of responsivity (R), the parameter that measures the sensitivity of a photoconductor to light, is below 1 AW(-1). We believe such poor performance is resulted from an intrinsic self-limited effect of present bare blend based device structure. Here we designed a PIN architecture for OTFPs, the PIN device exhibits a significantly improved high R value of 96.5 AW(-1). The PIN architecture and the performance the PIN device shows here should represent an important step in the development of OTFPs.

Applicability of the diffusion and simplified P3 theories for BWR pin-by-pin core analysis

International Nuclear Information System (INIS)

Tada, Kenichi; Yamamoto, Akio; Kitamura, Yasunori; Yamane, Yoshihiro; Watanabe, Masato; Noda, Hiroshi

2007-01-01

The pin-by-pin fine mesh core calculation method is considered as a candidate of next-generation core calculation method for BWR. In this study, the diffusion and the simplified P 3 (SP 3 ) theories are applied to the pin-by-pin core analysis of BWR. Performances of the diffusion and the SP 3 theories for cell-homogeneous pin-by-pin fine mesh BWR core analysis are evaluated through comparison with cell-heterogeneous detailed transport calculation by the method of characteristics (MOC). In this study, two-dimensional, 2x2 multi-assemblies geometry is used to compare the prediction accuracies of the diffusion and the SP 3 theories. The 2x2 multi- assemblies geometry consists of two types of 9x9 UO 2 assembly that have two different enrichment splittings. To mitigate the cell-homogenization error, the SPH method is applied for the pin-by-pin fine mesh calculation. The SPH method is a technique that reproduces a result of heterogeneous calculation by that of homogeneous calculation. The calculation results indicated that diffusion theory shows larger discrepancy than that of SP 3 theory on pin-wise fission rates. Furthermore, the accuracy of the diffusion theory would not be sufficient for the pin-by-pin fine mesh calculation. In contrast to the diffusion theory, the SP 3 theory shows much better accuracy on pin wise fission rates. Therefore, if the SP 3 theory is applied, the accuracy of the pin-by-pin fine mesh BWR core analysis will be higher and will be sufficient for production calculation. (author)

Comparison of SCDAP/RELAP5/MOD3 to TRAC-PF1/MOD1 for timing analysis of PWR fuel pin failures

International Nuclear Information System (INIS)

Jones, K.R.; Katsma, K.R.; Wade, N.L.; Siefken, L.J.; Straka, M.

1991-01-01

A comparison has been made of SCDAP/RELAP5/MOD3- and TRAC-PF1/MOD1- based calculations of the fuel pin failure timing (time from containment isolation signal to first fuel pin failure) in a loss-of-coolant accident (LOCA). The two codes were used to calculate the thermal-hydraulic boundary conditions for a complete, double-ended, offset-shear break of a cold leg in a Westinghouse 4-loop pressurized water reactor. Both calculations used the FRAPCON-2 code to calculate the steady-state fuel rod behavior and the FRAP-T6 code to calculate the transient fuel rod behavior. The analysis was performed for 16 combinations of fuel burnups and power peaking factors extending up to the Technical Specifications limits. While all calculations were made on a best-estimate basis, the SCDAP/RELAP5/MOD3 code has not yet been fully assessed for large-break LOCA analysis. The results indicate that SCDAP/RELAP5/MOD3 yields conservative fuel pin failure timing results in comparison to those generated using TRAC-PF1/MOD1. 7 refs., 5 figs

Pinning, flux diodes and ratchets for vortices interacting with conformal pinning arrays

International Nuclear Information System (INIS)

Olson Reichhardt, C. J.; Wang, Y. L.; Argonne National Laboratory; Xiao, Z. L.; Northern Illinois University, DeKalb, IL

2016-01-01

A conformal pinning array can be created by conformally transforming a uniform triangular pinning lattice to produce a new structure in which the six-fold ordering of the original lattice is conserved but where there is a spatial gradient in the density of pinning sites. Here we examine several aspects of vortices interacting with conformal pinning arrays and how they can be used to create a flux flow diode effect for driving vortices in different directions across the arrays. Under the application of an ac drive, a pronounced vortex ratchet effect occurs where the vortices flow in the easy direction of the array asymmetry. When the ac drive is applied perpendicular to the asymmetry direction of the array, it is possible to realize a transverse vortex ratchet effect where there is a generation of a dc flow of vortices perpendicular to the ac drive due to the creation of a noise correlation ratchet by the plastic motion of the vortices. We also examine vortex transport in experiments and compare the pinning effectiveness of conformal arrays to uniform triangular pinning arrays. In conclusion, we find that a triangular array generally pins the vortices more effectively at the first matching field and below, while the conformal array is more effective at higher fields where interstitial vortex flow occurs.

Excessive Cellular S-nitrosothiol Impairs Endocytosis of Auxin Efflux Transporter PIN2

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Min Ni

2017-11-01

Full Text Available S-nitrosoglutathione reductase (GSNOR1 is the key enzyme that regulates cellular levels of S-nitrosylation across kingdoms. We have previously reported that loss of GSNOR1 resulted in impaired auxin signaling and compromised auxin transport in Arabidopsis, leading to the auxin-related morphological phenotypes. However, the molecular mechanism underpinning the compromised auxin transport in gsnor1-3 mutant is still unknown. Endocytosis of plasma-membrane (PM-localized efflux PIN proteins play critical roles in auxin transport. Therefore, we investigate whether loss of GSNOR1 function has any effects on the endocytosis of PIN-FORMED (PIN proteins. It was found that the endocytosis of either the endogenous PIN2 or the transgenically expressed PIN2-GFP was compromised in the root cells of gsnor1-3 seedlings relative to Col-0. The internalization of PM-associated PIN2 or PIN2-GFP into Brefeldin A (BFA bodies was significantly reduced in gsnor1-3 upon BFA treatment in a manner independent of de novo protein synthesis. In addition, the exogenously applied GSNO not only compromised the endocytosis of PIN2-GFP but also inhibited the root elongation in a concentration-dependent manner. Taken together, our results indicate that, besides the reduced PIN2 level, one or more compromised components in the endocytosis pathway could account for the reduced endocytosis of PIN2 in gsnor1-3.

Investigation on macroscopic cross section model for BWR pin-by-pin core analysis - 118

International Nuclear Information System (INIS)

Fujita, T.; Tada, K.; Yamamoto, A.; Yamane, Y.; Kosaka, S.; Hirano, G.

2010-01-01

A cross section model used in the pin-by-pin core analysis for BWR is investigated. In the pin-by-pin core calculation method, pin-cell averaged cross sections are calculated for many combinations of state and history variables that have influences on the cross section and are tabulated prior to the core calculations. Variation of a cross section in a core simulator is classified into two different types, i.e., the instantaneous effect and the history effect. The instantaneous effect is incorporated by the variation of cross section which is caused by the instantaneous change of state variables. For this effect, the exposure, the void fraction, the fuel temperature, the moderator temperature and the control rod are used as indexes. The history effect is the cumulative effect of state variables. We treat this effect with a unified approach using the spectral history. To confirm accuracy of the cross section model, the pin-by-pin fission rate distribution and the k-infinity of fuel assembly which are obtained with the tabulated and the reference cross sections are compared. For the instantaneous effect, the present cross section model well reproduces the reference results for all off-nominal conditions. For the history effect, however, considerable differences both on the pin-by-pin fission rate distribution and the k-infinity are observed at high exposure points. (authors)

Transverse pinning versus intramedullary pinning in fifth metacarpal's neck fractures: A randomized controlled study with patient-reported outcome.

Science.gov (United States)

Galal, Sherif; Safwat, Wael

2017-01-01

The 5th metacarpal fractures accounts for 38% of all hand fractures given that the neck is the weakest point in metacarpals, so neck fracture is the most common metacarpal fracture. Surgical fixation is also advocated for such fractures to prevent mal-rotation of the little finger which will lead to fingers overlap in a clenched fist. Various methods are available for fixation of such fractures, like intramedullary & transverse pinning. There are very few reports in the literature comparing both techniques. Authors wanted to compare outcomes and complications of transverse pinning versus intramedullary pinning in fifth metacarpal's neck fractures. A single-center, parallel group, prospective, randomized study was conducted at an academic Level 1 Trauma Center from October 2014 to December 2016. A total of 80 patients with 5th metacarpal's neck fractures were randomized to pinning using either transverse pinning (group A) or intramedullary pinning (group B). Patients were assessed clinically on range of motion, patient-reported outcome using the Quick-DASH (Disabilities of the Arm, Shoulder, and Hand) questionnaire & radiographically. Two blinded observers assessed outcomes. At final follow up for each patient (12 months) the statistically significant differences were observed in operative time, the transverse pinning group showed shorter operative time, as well as complication rate as complications were observed only in intramedullary pinning group. No differences were found in range of motion or the Quick -DASH score. Both techniques are equally safe and effective treatment option for 5th metacarpal's neck fractures. The only difference was shorter operative time & less incidence of complications in transverse pinning group. Level II, Therapeutic study.

Clinical and histomorphometrical study on titanium dioxide-coated external fixation pins

Directory of Open Access Journals (Sweden)

Koseki H

2013-02-01

Full Text Available Hironobu Koseki,1 Tomohiko Asahara,1 Takayuki Shida,1 Itaru Yoda,1 Hidehiko Horiuchi,1 Koumei Baba,2 Makoto Osaki11Department of Orthopedic Surgery, Graduate School of Medicine, Nagasaki University, 2Industrial Technology Center of Nagasaki, Nagasaki, JapanBackground: Pin site infection is the most common and significant complication of external fixation. In this work, the efficacy of pins coated with titanium dioxide (TiO2 for inhibition of infection was compared with that of stainless steel control pins in an in vivo study.Methods: Pins contaminated with an identifiable Staphylococcus aureus strain were inserted into femoral bone in a rat model and exposed to ultraviolet A light for 30 minutes. On day 14, the animals were sacrificed and the bone and soft tissue around the pin were retrieved. The clinical findings and histological findings were evaluated in 60 samples.Results: Clinical signs of infection were present in 76.7% of untreated pins, but in only 36.7% of TiO2-coated pins. The histological bone infection score and planimetric rate of occupation for bacterial colonies and neutrophils in the TiO2-coated pin group were lower than those in the control group. The bone-implant contact ratio of the TiO2-coated pin group was significantly higher (71.4% than in the control pin group (58.2%. The TiO2 was successful in decreasing infection both clinically and histomorphometrically.Conclusion: The photocatalytic bactericidal effect of TiO2 is thought to be useful for inhibiting pin site infection after external fixation.Keywords: titanium dioxide, external fixation, bactericidal activity, Staphylococcus aureus

Crystallization and preliminary X-ray diffraction studies of l-rhamnose isomerase from Pseudomonas stutzeri

International Nuclear Information System (INIS)

Yoshida, Hiromi; Wayoon, Poonperm; Takada, Goro; Izumori, Ken; Kamitori, Shigehiro

2006-01-01

Recombinant l-rhamnose isomerase from P. stutzeri has been crystallized. Diffraction data have been collected to 2.0 Å resolution. l-Rhamnose isomerase from Pseudomonas stutzeri (P. stutzeril-RhI) catalyzes not only the reversible isomerization of l-rhamnose to l-rhamnulose, but also isomerization between various rare aldoses and ketoses. Purified His-tagged P. stutzeril-RhI was crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the monoclinic space group P2 1 , with unit-cell parameters a = 74.3, b = 104.0, c = 107.0 Å, β = 106.8°. Diffraction data have been collected to 2.0 Å resolution. The molecular weight of the purified P. stutzeril-RhI with a His tag at the C-terminus was confirmed to be 47.7 kDa by MALDI–TOF mass-spectrometric analysis and the asymmetric unit is expected to contain four molecules

A preliminary X-ray study of sedoheptulose-7-phosphate isomerase from Burkholderia pseudomallei

International Nuclear Information System (INIS)

Kim, Mi-Sun; Shin, Dong Hae

2009-01-01

Sedoheptulose-7-phosphate isomerase (GmhA) from B. pseudomallei is one of the targets of antibiotic adjuvants for melioidosis. In this study, GmhA has been cloned, expressed, purified and crystallized. Sedoheptulose-7-phosphate isomerase (GmhA) converts d-sedoheptulose 7-phosphate to d,d-heptose 7-phosphate. This is the first step in the biosynthesis pathway of NDP-heptose, which is responsible for the pleiotropic phenotype. This biosynthesis pathway is the target of inhibitors to increase the membrane permeability of Gram-negative pathogens or of adjuvants working synergistically with known antibiotics. Burkholderia pseudomallei is the causative agent of melioidosis, a seriously invasive disease in animals and humans in tropical and subtropical areas. GmhA from B. pseudomallei is one of the targets of antibiotic adjuvants for melioidosis. In this study, GmhA has been cloned, expressed, purified and crystallized. Synchrotron X-ray data were also collected to 1.9 Å resolution. The crystal belonged to the primitive orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 61.3, b = 84.2, c = 142.3 Å. A full structural determination is under way in order to provide insights into the structure–function relationships of this protein

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

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.

Cyclophilin B facilitates the replication of Orf virus

OpenAIRE

Zhao, Kui; Li, Jida; He, Wenqi; Song, Deguang; Zhang, Ximu; Zhang, Di; Zhou, Yanlong; Gao, Feng

2017-01-01

Background Viruses interact with host cellular factors to construct a more favourable environment for their efficient replication. Expression of cyclophilin B (CypB), a cellular peptidyl-prolyl cis-trans isomerase (PPIase), was found to be significantly up-regulated. Recently, a number of studies have shown that CypB is important in the replication of several viruses, including Japanese encephalitis virus (JEV), hepatitis C virus (HCV) and human papillomavirus type 16 (HPV 16). However, the f...

Triosephosphate isomerase: energetics of the reaction catalyzed by the yeast enzyme expressed in Escherichia coli

International Nuclear Information System (INIS)

Nickbarg, E.B.; Knowles, J.R.

1988-01-01

Triosephosphate isomerase from bakers' yeast, expressed in Escherichia coli strain DF502(p12), has been purified to homogeneity. The kinetics of the reaction in each direction have been determined at pH 7.5 and 30 degrees C. Deuterium substitution at the C-2 position of substrate (R)-glyceraldehyde phosphate and at the 1-pro-R position of substrate dihydroxyacetone phosphate results in kinetic isotope effects on kcat of 1.6 and 3.4, respectively. The extent of transfer of tritium from [1(R)- 3 H]dihydroxyacetone phosphate to product (R)-glyceraldehyde phosphate during the catalyzed reaction is only 3% after 66% conversion to product, indicating that the enzymic base that mediates proton transfer is in rapid exchange with solvent protons. When the isomerase-catalyzed reaction is run in tritiated water in each direction, radioactivity is incorporated both into the remaining substrate and into the product. In the exchange-conversion experiment with dihydroxyacetone phosphate as substrate, the specific radioactivity of remaining dihydroxyacetone phosphate rises as a function of the extent of reaction with a slope of about 0.3, while the specific radioactivity of the products is 54% that of the solvent. In the reverse direction with (R)-glyceraldehyde phosphate as substrate, the specific radioactivity of the product formed is only 11% that of the solvent, while the radioactivity incorporated into the remaining substrate (R)-glyceraldehyde phosphate also rises as a function of the extent of reaction with a slope of 0.3. These results have been analyzed according to the protocol described earlier to yield the free energy profile of the reaction catalyzed by the yeast isomerase

Preliminary crystallographic analysis of two hypothetical ribose-5-phosphate isomerases from Streptococcus mutans

International Nuclear Information System (INIS)

Wang, Chen; Fan, Xuexin; Cao, Xiaofang; Liu, Xiang; Li, Lanfen; Su, Xiaodong

2012-01-01

Two hypothetical ribose-5-phosphate isomerases from S. mutans have been produced in E. coli and crystallized. The crystals diffracted to high resolutions suitable for crystallographic analyses. Study of the enzymes from sugar metabolic pathways may provide a better understanding of the pathogenesis of the human oral pathogen Streptococcus mutans. Bioinformatics, biochemical and crystallization methods were used to characterize and understand the function of two putative ribose-5-phosphate isomerases: SMU1234 and SMU2142. The proteins were cloned and constructed with N-terminal His tags. Protein purification was performed by Ni 2+ -chelating and size-exclusion chromatography. The crystals of SUM1234 diffracted to 1.9 Å resolution and belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 48.97, b = 98.27, c = 101.09 Å, α = β = γ = 90°. The optimized SMU2142 crystals diffracted to 2.7 Å resolution and belonged to space group P1, with unit-cell parameters a = 53.7, b = 54.1, c = 86.5 Å, α = 74.2, β = 73.5, γ = 83.7°. Initial phasing of both proteins was attempted by molecular replacement; the structure of SMU1234 could easily be solved, but no useful results were obtained for SMU2142. Therefore, SeMet-labelled SMU2142 will be prepared for phasing

Structural insights from a novel invertebrate triosephosphate isomerase from Litopenaeus vannamei

Science.gov (United States)

Lopez-Zavala, Alonso A.; Carrasco-Miranda, Jesus S.; Ramirez-Aguirre, Claudia D.; López-Hidalgo, Marisol; Benitez-Cardoza, Claudia G.; Ochoa-Leyva, Adrian; Cardona-Felix, Cesar S.; Diaz-Quezada, Corina; Rudiño-Piñera, Enrique; Sotelo-Mundo, Rogerio R.; Brieba, Luis G.

2016-01-01

Triosephosphate isomerase (TIM; EC 5.3.1.1) is a key enzyme involved in glycolysis and gluconeogenesis. Glycolysis is one of the most regulated metabolic pathways, however little is known about the structural mechanisms for its regulation in non-model organisms, like crustaceans. To understand the structure and function of this enzyme in invertebrates, we obtained the crystal structure of triosephosphate isomerase from the marine Pacific whiteleg shrimp (Litopenaeus vannamei, LvTIM) in complex with its inhibitor 2-phosphogyceric acid (2-PG) at 1.7 Å resolution. LvTIM assembles as a homodimer with residues 166-176 covering the active site and residue Glu166 interacting with the inhibitor. We found that LvTIM is the least stable TIM characterized to date, with the lowest range of melting temperatures, and with the lowest activation enthalpy associated with the thermal unfolding process reported. In TIMs dimer stabilization is maintained by an interaction of loop 3 by a set of hydrophobic contacts between subunits. Within these contacts, the side chain of a hydrophobic residue of one subunit fits into a cavity created by a set of hydrophobic residues in the neighboring subunit, via a "ball and socket" interaction. LvTIM presents a Cys47 at the "ball" inter-subunit contact indicating that the character of this residue is responsible for the decrease in dimer stability. Mutational studies show that this residue plays a role in dimer stability but is not a solely determinant for dimer formation. PMID:27614148

Defect pin behaviour in the DFR

International Nuclear Information System (INIS)

Sloss, W.M.; Bagley, K.Q.; Edmonds, E.; Potter, P.E.

1979-01-01

A program of defective fuel pin irradiations has been carried out in the DFR. This program employed fuel pins which had failed during previous irradiations (natural defects) and pins in which simulated failures (artificial defects) had been induced prior to irradiation or during an intermediate examination stage at moderate or substantial burnups. The artificial defects simulated longitudinal ruptures and were normally located at positions near the top, middle and bottom of the pin where clad temperatures were 450, 540 and 630 0 C respectively. The fuel was mixed U-Pu oxide, and fuel form, stoichiometry, clad type, pin diameter, linear rating, and burnup were among the variables examined. The defect pin tests were normally carried out in single pin or trefoil type vehicles. After irradiation all the pins were subjected to the normal nondestructive examination procedures and the visual, radiographic, gamma-scanning, and dimensional change results are presented. Several pins were destructively examined and the metallographic data are discussed

Obtaining mutants of Streptomyces griseoflavus strain 1339, producers of glucose isomerase, following gamma irradiation

International Nuclear Information System (INIS)

Dzhedzheva, G.; Stoeva, N.; Stojchev, M.

1990-01-01

A water suspension of Streptomyces griseoflavus strain 1339 spores of a density of 8.7.10 6 spores/cm 3 is gamma irradiated ( 60 Co, RHM-γ-20, 30.3 Gy/min). The survival of Streptomyces griseoflavus strain 1339 spores was determined depending on radiation doses, exposure times and incubation temperature. Five major morphological types of colonies were isolated, characterized by different levels of glucose isomerase activity. Maximum specific glucose isomerase activity (GIU/g) was attained after the third gamma irradiation step using a dose of 3000 Gy. 2 tabs., 3 figs., 7 refs

Flux pinning characteristics of YBCO coated conductor

International Nuclear Information System (INIS)

Matsushita, T.; Watanabe, T.; Fukumoto, Y.; Yamauchi, K.; Kiuchi, M.; Otabe, E.S.; Kiss, T.; Watanabe, T.; Miyata, S.; Ibi, A.; Muroga, T.; Yamada, Y.; Shiohara, Y.

2005-01-01

Flux pinning properties of PLD-processed YBCO coated conductors deposited on IBAD substrate are investigated. The thickness of YBCO layer is changed in the range of 0.27-1.0 μm. The thickness dependence of critical current density, n-value and irreversibility field are measured in a wide range of magnetic field. The results are compared with the theoretical flux creep-flow model. It is found that these pinning properties are strongly influenced by the thickness as well as the pinning strength. Optimum condition for high field application of this superconductor is discussed

Roles of pinning strength and density in vortex melting

International Nuclear Information System (INIS)

Obaidat, I M; Khawaja, U Al; Benkraouda, M

2008-01-01

We have investigated the role of pinning strength and density on the equilibrium vortex-lattice to vortex-liquid phase transition under several applied magnetic fields. This study was conducted using a series of molecular dynamic simulations on several samples with different strengths and densities of pinning sites which are arranged in periodic square arrays. We have found a single solid-liquid vortex transition when the vortex filling factor n>1. We have found that, for fixed pinning densities and strengths, the melting temperature, T m , decreases almost linearly with increasing magnetic field. Our results provide direct numerical evidence for the significant role of both the strength and density of pinning centers on the position of the melting line. We have found that the vortex-lattice to vortex-liquid melting line shifts up as the pinning strength or the pinning density was increased. The effect on the melting line was found to be more pronounced at small values of strength and density of pinning sites

Improved pinning by multiple in-line damage

Energy Technology Data Exchange (ETDEWEB)

Weinstein, Roy [Beam Particle Dynamics Laboratories, University of Houston, Houston, TX 77204-5005 (United States); Sawh, Ravi-Persad [Beam Particle Dynamics Laboratories, University of Houston, Houston, TX 77204-5005 (United States); Gandini, Alberto [Beam Particle Dynamics Laboratories, University of Houston, Houston, TX 77204-5005 (United States); Parks, Drew [Beam Particle Dynamics Laboratories, University of Houston, Houston, TX 77204-5005 (United States)

2005-02-01

Columnar pinning centres provide the largest pinning potential, U{sub pin}, but not the greatest J{sub c} or pinnable field, B{sub pin}. Characteristics of ion-generated columnar defects which limit J{sub c} and B{sub pin} are discussed, including reduction of the percolation path, and the need for a larger number of columns of damage, for pinning, than are usually estimated. It is concluded that columnar pinning centres limit B{sub pin} to less than 4 T, and also severely reduce J{sub c}. The goal of maximizing U{sub pin}, via columnar centres, appears to have obscured a more rewarding approach and resulted in neglect of a large regime of ion interactions. Evidence is reviewed that multiple in-line damage (MILD), described herein, can provide orders of magnitude higher J{sub c} and B{sub pin}, despite providing lower U{sub pin}. The MILD pinning centre morphology is discussed, and it is estimated that for present-day large grain high T{sub c} superconductors, a J{sub c} value of {approx}10{sup 6}Acm{sup -2} is obtainable at 77 K, even when crystal plane alignment and weak links are not improved. In addition, the pinned field is increased by over an order of magnitude. An experiment is proposed to confirm these calculations, directly compare MILD pinning to continuous columnar pinning, and determine the optimum MILD structure. Applications of MILD pinning are discussed.

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.

Displaced humeral lateral condyle fractures in children: should we bury the pins?

Science.gov (United States)

Das De, Soumen; Bae, Donald S; Waters, Peter M

2012-09-01

The purpose of this investigation was to determine if leaving Kirschner wires exposed is more cost-effective than burying them subcutaneously after open reduction and internal fixation (ORIF) of humeral lateral condyle fractures. A retrospective cohort study of all lateral condyle fractures treated over a 10-year period at a single institution was performed. Data on surgical technique, fracture healing, and complications were analyzed, as well as treatment costs. A decision analysis model was then constructed to compare the strategies of leaving the pins exposed versus buried. Finally, sensitivity analyses were performed, assessing cost-effectiveness when infection rates and costs of treating deep infections were varied. A total of 235 children with displaced fractures were treated with ORIF using Kirschner wires. Pins were left exposed in 41 cases (17.4%) and buried in 194 cases (82.6%); the age, sex, injury mechanisms, and fracture patterns were similar in both the groups. The median time to removal of implants was shorter with exposed versus buried pins (4 vs. 6 wk, Pfracture union or loss of reduction rates. The rate of superficial infection was higher with exposed pins (9.8% vs. 3.1%), but this was not statistically significant (P=0.076). There were no deep infections with exposed pins, whereas the rate of deep infection was 0.5% with buried pins (P=1.00). Buried pins were associated with additional complications, including symptomatic implants (7.2%); pins protruding through the skin (16%); internal pin migration necessitating additional surgery (1%); and skin necrosis (1%). The decision analysis revealed that leaving pins exposed resulted in an average cost savings of $3442 per patient. This strategy remained cost-effective even when infection rates with exposed pins approached 40%. Leaving the pins exposed after ORIF of lateral condyle fractures is safe and more cost-effective than burying the pins subcutaneously. Retrospective cohort study (level III).

Fuel pin transfer tool

International Nuclear Information System (INIS)

Patenaude, R. S.

1985-01-01

A fuel pin transfer tool has a latching device of the collet type attached to a first member movable vertically through a long work stroke enabling a fuel pin in an under water assembly to be engaged and withdrawn therefrom or placed therein and released. The latching device has a collet provided with a plurality of resilient fingers having cam portions normally spaced apart to receive the upper end of a fuel pin between them and a second member, movable vertically through a short stroke relative to the first member is provided with cam portions engageable with those of the fingers and is yieldably and resiliently held in a raised position in which its cam portions engage those of the fingers and force the fingers into their pin-gripping positions. When a predetermined force is applied to the second member, it is so moved that its cam portions are disengaged from the cam portions of the fingers permitting the latter to move into their normal relationship in which a gripped pin is released or another pin received but with their pin-gripping relationship positively re-established and maintained once the force on the tubular member is lessened. Movement of the first member in either direction and movement of the second member into its raised position is attended by forces inadequate to affect the integrity of fuel pin cladding. That force is applied in the preferred embodiment, by a power operated actuator which is within the upper portion of a housing and, in the preferred embodiment, carried by the long stroke member but always in the upper housing portion which is of a material sufficiently translucent to enable the actuator to be observed throughout the work stroke and is sufficiently light in weight to prevent the tool from being top heavy

Results of transient overpower events on breached and unbreached fuel pins

International Nuclear Information System (INIS)

Strain, R.V.; Tsai, H.C.; Neimark, L.A.; Aratani, K.

1986-04-01

The objective of the extended overpower tests on intact pins was to determine the pin cladding breaching thresholds vis-a-vis the Plant Protection System (PPS) trip settings, typically at ∼10 to 15% overpower. These tests emphasize slow operational-type transients in light of earlier work which suggested that irradiated mixed-oxide fuel pins may be particularly vulnerable in the slow ramp-rate regime. An overview of the extended overpower test series was previously reported. More recent results on two of the tests in this series are included in this paper. These two tests, designated TOPI-1A and TOPI-1B, were each conducted on a 19-pin assembly with various pin design, operation and burnup variables. The overpower ramp rates for the TOPI-1A and -1B tests were 0.1%/s and 10%/s, respectively

Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA

International Nuclear Information System (INIS)

Ravaud, Stéphanie; Watzlawick, Hildegard; Haser, Richard; Mattes, Ralf; Aghajari, Nushin

2005-01-01

The P. rubrum sucrose isomerase SmuA, a key enzyme in the industrial production of isomaltulose, was crystallized and diffraction data were collected to 1.95 Å resolution. Palatinose (isomaltulose, α-d-glucosylpyranosyl-1,6-d-fructofuranose), a nutritional and acariogenic reducing sugar, is industrially obtained from sucrose by using immobilized cells of Protaminobacter rubrum that produce the sucrose isomerase SmuA. The isomerization of sucrose catalyzed by this enzyme also results in the formation of trehalulose (α-d-glucosylpyranosyl-1,1-d-fructofuranose) in smaller amounts and glucose, fructose and eventually isomaltose as by-products, which lower the yield of the reaction and complicate the recovery of palatinose. The determination of the three-dimensional structure of SmuA will provide a basis for rational protein-engineering studies in order to optimize the industrial production of palatinose. A recombinant form of the 67.3 kDa SmuA enzyme has been crystallized in the native state by the vapour-diffusion method. Crystals belong to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 61.6, b = 81.4, c = 135.6 Å, and diffract to 1.95 Å resolution on a synchrotron-radiation source

Crystallization and preliminary X-ray crystallographic analysis of l-rhamnose isomerase with a novel high thermostability from Bacillus halodurans

International Nuclear Information System (INIS)

Doan, Thi-Ngoc-Thanh; Prabhu, Ponnandy; Kim, Jin-Kwang; Ahn, Yeh-Jin; Natarajan, Sampath; Kang, Lin-Woo; Park, Geon Tae; Lim, Sang-Boem; Lee, Jung-Kul

2010-01-01

l-Rhamnose isomerase (l-RhI) from B. halodurans has been purified and crystallized. The crystals of l-RhI belonged to the monoclinic space group P2 1 , with unit-cell parameters a = 83.2, b = 164.9, c = 92.0 Å, β = 116.0°, and diffracted to 2.5 Å resolution. l-Rhamnose isomerases catalyze isomerization between l-rhamnose (6-deoxy-l-mannose) and l-rhamnulose (6-deoxy-l-fructose), which is the first step in rhamnose catabolism. l-Rhamnose isomerase from Bacillus halodurans ATCC BAA-125 (BHRI) exhibits interesting characteristics such as high thermostability and selective substrate specificity. BHRI fused with an HHHHHH sequence was purified and crystallized in order to elucidate the molecular basis of its unique enzymatic properties. The crystals were grown by the hanging-drop vapour-diffusion method and belonged to the monoclinic space group P2 1 , with unit-cell parameters a = 83.2, b = 164.9, c = 92.0 Å, β = 116.0°. Diffraction data were collected to 2.5 Å resolution. According to a Matthews coefficient calculation, there are four monomers in the asymmetric unit with a V M of 3.0 Å 3 Da −1 and a solvent content of 59.3%. The initial structure of BHRI has been determined by the molecular-replacement method

Genomic analysis of a xylose operon and characterization of novel xylose isomerase and xylulokinase from Bacillus coagulans NL01.

Science.gov (United States)

Zheng, Zhaojuan; Lin, Xi; Jiang, Ting; Ye, Weihua; Ouyang, Jia

2016-08-01

To investigate the xylose operon and properties of xylose isomerase and xylulokinase in Bacillus coagulans that can effectively ferment xylose to lactic acid. The xylose operon is widely present in B. coagulans. It is composed of four putative ORFs. Novel xylA and xylB from B. coagulans NL01 were cloned and expressed in Escherichia coli. Sequence of xylose isomerase was more conserved than that of xylulokinase. Both the enzymes exhibited maximum activities at pH 7-8 but with a high temperature maximum of 80-85 °C, divalent metal ion was prerequisite for their activation. Xylose isomerase and xylulokinase were most effectively activated by Ni(2+) and Co(2+), respectively. Genomic analysis of xylose operon has contributed to understanding xylose metabolism in B. coagulans and the novel xylose isomerase and xylulokinase might provide new alternatives for metabolic engineering of other strains to improve their fermentation performance on xylose.

Functionally different PIN proteins control auxin flux during bulbil development in Agave tequilana.

Science.gov (United States)

Abraham Juárez, María Jazmín; Hernández Cárdenas, Rocío; Santoyo Villa, José Natzul; O'Connor, Devin; Sluis, Aaron; Hake, Sarah; Ordaz-Ortiz, José; Terry, Leon; Simpson, June

2015-07-01

In Agave tequilana, reproductive failure or inadequate flower development stimulates the formation of vegetative bulbils at the bracteoles, ensuring survival in a hostile environment. Little is known about the signals that trigger this probably unique phenomenon in agave species. Here we report that auxin plays a central role in bulbil development and show that the localization of PIN1-related proteins is consistent with altered auxin transport during this process. Analysis of agave transcriptome data led to the identification of the A. tequilana orthologue of PIN1 (denoted AtqPIN1) and a second closely related gene from a distinct clade reported as 'Sister of PIN1' (denoted AtqSoPIN1). Quantitative real-time reverse transcription-PCR (RT-qPCR) analysis showed different patterns of expression for each gene during bulbil formation, and heterologous expression of the A. tequilana PIN1 and SoPIN1 genes in Arabidopsis thaliana confirmed functional differences between these genes. Although no free auxin was detected in induced pedicel samples, changes in the levels of auxin precursors were observed. Taken as a whole, the data support the model that AtqPIN1 and AtqSoPIN1 have co-ordinated but distinct functions in relation to auxin transport during the initial stages of bulbil formation. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Complications after pinning of supracondylar distal humerus fractures.

Science.gov (United States)

Bashyal, Ravi K; Chu, Jennifer Y; Schoenecker, Perry L; Dobbs, Matthew B; Luhmann, Scott J; Gordon, J Eric

2009-01-01

Supracondylar distal humerus fractures are one of the most common skeletal injuries in children. The current treatment of choice in North America is closed reduction and percutaneous pin fixation. Often surgeons leave the pins exposed beneath a cast but outside the skin. Great variation exists with respect to preoperative skin preparation, and perioperative antibiotic administration. Few data exist regarding the rate of infection and other complications. The purpose of this study is to review a large series of children to evaluate the rate of infection and other complications. A retrospective review was carried out of all patients treated at our institution over an 11-year period. A total of 622 patients were identified that were followed for a minimum of 2 weeks after pin removal. Seventeen patients had flexion-type fractures, 294 had type II fractures, and 311 had type III fractures. Seventy-four fractures (11.9%) had preoperative nerve deficits with anterior interosseous palsies being the most common (33 fractures, 5.3%). Preoperative antibiotics were given to 163 patients (26.2%). Spray and towel draping were used in 362 patients, paint and towel draping were used in 65 patients, alcohol paint and towel draping were used in 146 patients, and a full preparation and draping were used in 13 patients. The pins were left exposed under the cast in 591 fractures (95%), and buried beneath the skin in 31 fractures (5.0%). A medial pin was placed in 311 fractures with a small incision made to aid placement in 18 of these cases. The most common complication was pin migration necessitating unexpected return to the operating room for pin removal in 11 patients (1.8%). One patient developed a deep infection with septic arthritis and osteomyelitis (0.2%). Five additional patients had superficial skin infections and were treated with oral antibiotics for a total infection rate of 6 of 622 patients (1.0%). One patient ultimately had a malunion and 4 others returned to the

PROCOPE, Collision Probability in Pin Clusters and Infinite Rod Lattices

International Nuclear Information System (INIS)

Amyot, L.; Daolio, C.; Benoist, P.

1984-01-01

1 - Nature of physical problem solved: Calculation of directional collision probabilities in pin clusters and infinite rod lattices. 2 - Method of solution: a) Gauss integration of analytical expressions for collision probabilities. b) alternately, an approximate closed expression (not involving integrals) may be used for pin-to-pin interactions. 3 - Restrictions on the complexity of the problem: number of fuel pins must be smaller than 62; maximum number of groups of symmetry is 300

High-performance whole core Pin-by-Pin calculation based on EFEN-SP_3 method

International Nuclear Information System (INIS)

Yang Wen; Zheng Youqi; Wu Hongchun; Cao Liangzhi; Li Yunzhao

2014-01-01

The EFEN code for high-performance PWR whole core pin-by-pin calculation based on the EFEN-SP_3 method can be achieved by employing spatial parallelization based on MPI. To take advantage of the advanced computing and storage power, the entire problem spatial domain can be appropriately decomposed into sub-domains and the assigned to parallel CPUs to balance the computing load and minimize communication cost. Meanwhile, Red-Black Gauss-Seidel nodal sweeping scheme is employed to avoid the within-group iteration deterioration due to spatial parallelization. Numerical results based on whole core pin-by-pin problems designed according to commercial PWRs demonstrate the following conclusions: The EFEN code can provide results with acceptable accuracy; Communication period impacts neither the accuracy nor the parallel efficiency; Domain decomposition methods with smaller surface to volume ratio leads to greater parallel efficiency; A PWR whole core pin-by-pin calculation with a spatial mesh 289 × 289 × 218 and 4 energy groups could be completed about 900 s by using 125 CPUs, and its parallel efficiency is maintained at about 90%. (authors)

Incidence and risk factors for pin tract infection in external fixation of ...

African Journals Online (AJOL)

Incidence and risk factors for pin tract infection in external fixation of fractures ... for pin tract infection, there were 93 pins scored grade 1, 32 pins grade 2, 15 ... The incidence increased from 20.5% in closed fractures to 75.9% in open fractures.

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...

A comparative CFD investigation of helical wire-wrapped 7, 19 and 37 fuel pin bundles and its extendibility to 217 pin bundle

International Nuclear Information System (INIS)

Gajapathy, R.; Velusamy, K.; Selvaraj, P.; Chellapandi, P.; Chetal, S.C.

2009-01-01

Preliminary investigations of sodium flow and temperature distributions in heat generating fuel pin bundles with helical spacer wires have been carried out. Towards this, the 3D conservation equations of mass, momentum and energy have been solved using a commercial computational fluid dynamics (CFD) code. Turbulence has been accounted through the use of high Reynolds number version of standard k-ε model, with uniform mesh density respecting wall function requirements. The geometric details of the bundle and the heat flux in are similar to that of the Indian Prototype Fast Breeder Reactor (PFBR) that is currently under construction. The mixing characteristics of the flow among the peripheral and central zones are compared for 7, 19 and 37 fuel pin bundles and the characteristics are extended to a 217 pin bundle. The friction factors of the pin bundles obtained from the present study is seen to agree well with the values derived from experimental correlations. It is found that the normalized outlet velocities in the peripheral and central zones are nearly equal to 1.1-0.9, respectively which is in good agreement with the published hydraulic experimental measurements of 1.1-0.85 for a 91 pin bundle. The axial velocity is the maximum in the peripheral zone where spacer wires are located and minimum in the zones which are diametrically opposite to the respective zone of maximum velocity. The sodium temperature is higher in the zones where the flow area and mass flow rates are less due to the presence of the spacer wires though the axial velocity is higher there. It is the minimum in the peripheral zones where the circumferential flow is larger. Based on the flow and temperature distributions obtained for 19 and 37 pin bundles, a preliminary extrapolation procedure has been established for estimating the temperatures of peripheral and central zones of 217 pin bundle.

MICROCONTROLLER PIN CONFIGURATION TOOL

OpenAIRE

Bhaskar Joshi; F. Mohammed Rizwan; Dr. Rajashree Shettar

2012-01-01

Configuring the micro controller with large number of pins is tedious. Latest Infine on microcontroller contains more than 200 pins and each pin has classes of signals. Therefore the complexity of the microcontroller is growing. It evolves looking into thousands of pages of user manual. For a user it will take days to configure the microcontroller with the peripherals. We need an automated tool to configure the microcontroller so that the user can configure the microcontroller without having ...

MONJU fuel pin performance analysis

International Nuclear Information System (INIS)

Kitagawa, H.; Yamanaka, T.; Hayashi, H.

1979-01-01

Monju fuel pin has almost the same properties as other LMFBR fuel pins, i.e. Phenix, PFR, CRBR, but would be irradiated under severe conditions: maximum linear heat rate of 381 watt/cm, hot spot cladding temperature of 675 deg C, peak burnup of 131,000 MWd/t, peak fluence (E greater than 0.1 MeV) of 2.3 10 23 n/cm 2 . In order to understand in-core performance of Monju fuel pin, its thermal and mechanical behaviour was predicted using the fast running performance code SIMPLE. The code takes into account pellet-cladding interaction due to thermal expansion and swelling, gap conductance, structural changes of fuel pellets, fission product gas release with burnup and temperature increase, swelling and creep of fuel pellets, corrosion of cladding due to sodium flow and chemical attack by fission products, and cumulative damage of the cladding due to thermal creep

Stress relaxation of thermally bowed fuel pins

International Nuclear Information System (INIS)

Crossland, I.G.; Speight, M.V.

1983-01-01

The presence of cross-pin temperature gradients in nuclear reactor fuel pins produces differential thermal expansion which, in turn, causes the fuel pin to bow elastically. If the pin is restrained in any way, such thermal bowing causes the pin to be stressed. At high temperatures these stresses can relax by creep and it is shown here that this causes the pin to suffer an additional permanent deflection, so that when the cross-pin temperature difference is removed the pin remains bowed. By representing the cylindrical pin by an equivalent I-beam, the present work examines this effect when it takes place by secondary creep. Two restraint systems are considered, and it is demonstrated that the rate of relaxation depends mainly upon the creep equation, and hence the temperature, and also the magnitude of the initial stresses. (author)

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

ITP Hanford Type 40 pin electrical connector failure analysis

International Nuclear Information System (INIS)

Imrich, K.J.

1993-01-01

Corrosion products observed on the ITP Hanford Type 40 pin electrical connectors would be expected to adversely affect the power and control signals supplied to process equipment in the filter cell by the connectors. Corrosion products were consistent with those found on similar pins in DWPF. The recommendations based on the findings in this investigation are as follows: (1) Replace male and female rhodium plated pins with gold plated pins. (2) Replace the galvanized carbon steel spring on the male connector with a stainless steel spring. (3) Install protective caps over Hanford connectors when jumpers are removed

[Deficiency of triosephosphate isomerase. Apropos of 2 new cases].

Science.gov (United States)

Delso Martínez, M C; Uriel Miñana, P; Pérez Lugmus, G; Giménez Mas, J A; Baldellou Vázquez, A

1983-08-01

Two siblings, born of a no consanguineous couple, a female and a male, affected by a severe and progressive neurological disease and chronic hemolytic anemia are presented. Their clinical, hematological, biochemical and pathological studies are discussed. One of the patients showed a triosephosphate isomerase deficiency and the carrier condition of their parents was tested. Commentaries about physiopathology of this disease are made.

Automated system for loading nuclear fuel pins

International Nuclear Information System (INIS)

Marshall, J.L.

1983-10-01

A completely automatic and remotely controlled fuel pin fabrication system is being designed by the Westinghouse Hanford Company. The Pin Operations System will produce fuel pins for the Fast Flux Test Facility (FFTF) and the Clinch River Breeder Reactor Plant (CRBRP). The system will assemble fuel pin components into cladding tubes in a controlled environment. After fuel loading, the pins are filled with helium, the tag gas capsules are inserted, and the top end cap welded. Following welding, the pins are surveyed to assure they are free of contamination and then the pins are helium leak tested

Fabrication of oxide dispersion strengthened ferritic clad fuel pins

International Nuclear Information System (INIS)

Zirker, L.R.; Bottcher, J.H.; Shikakura, S.; Tsai, C.L.

1991-01-01

A resistance butt welding procedure was developed and qualified for joining ferritic fuel pin cladding to end caps. The cladding are INCO MA957 and PNC ODS lots 63DSA and 1DK1, ferritic stainless steels strengthened by oxide dispersion, while the end caps are HT9 a martensitic stainless steel. With adequate parameter control the weld is formed without a residual melt phase and its strength approaches that of the cladding. This welding process required a new design for fuel pin end cap and weld joint. Summaries of the development, characterization, and fabrication processes are given for these fuel pins. 13 refs., 6 figs., 1 tab

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...

Pin Component Technology (V1.0) and Its C Interface

National Research Council Canada - National Science Library

Hissam, Scott; Ivers, James; Plakosh, Daniel; Wallnau, Kurt C

2005-01-01

.... Pin implements the container idiom for software components. Containers provide a pre-fabricated "shell" in which custom code executes and through which all interactions between custom code and its external environment are mediated...

A proposed parameterization of interface discontinuity factors depending on neighborhood for pin-by-pin diffusion computations for LWR

International Nuclear Information System (INIS)

Herrero, Jose Javier; Garcia-Herranz, Nuria; Ahnert, Carol

2011-01-01

There exists an interest in performing full core pin-by-pin computations for present nuclear reactors. In such type of problems the use of a transport approximation like the diffusion equation requires the introduction of correction parameters. Interface discontinuity factors can improve the diffusion solution to nearly reproduce a transport solution. Nevertheless, calculating accurate pin-by-pin IDF requires the knowledge of the heterogeneous neutron flux distribution, which depends on the boundary conditions of the pin-cell as well as the local variables along the nuclear reactor operation. As a consequence, it is impractical to compute them for each possible configuration. An alternative to generate accurate pin-by-pin interface discontinuity factors is to calculate reference values using zero-net-current boundary conditions and to synthesize afterwards their dependencies on the main neighborhood variables. In such way the factors can be accurately computed during fine-mesh diffusion calculations by correcting the reference values as a function of the actual environment of the pin-cell in the core. In this paper we propose a parameterization of the pin-by-pin interface discontinuity factors allowing the implementation of a cross sections library able to treat the neighborhood effect. First results are presented for typical PWR configurations. (author)

A proposed parameterization of interface discontinuity factors depending on neighborhood for pin-by-pin diffusion computations for LWR

Energy Technology Data Exchange (ETDEWEB)

Herrero, Jose Javier; Garcia-Herranz, Nuria; Ahnert, Carol, E-mail: herrero@din.upm.es, E-mail: nuria@din.upm.es, E-mail: carol@din.upm.es [Departamento de Ingenieria Nuclear, Universidad Politecnica de Madrid (Spain)

2011-07-01

There exists an interest in performing full core pin-by-pin computations for present nuclear reactors. In such type of problems the use of a transport approximation like the diffusion equation requires the introduction of correction parameters. Interface discontinuity factors can improve the diffusion solution to nearly reproduce a transport solution. Nevertheless, calculating accurate pin-by-pin IDF requires the knowledge of the heterogeneous neutron flux distribution, which depends on the boundary conditions of the pin-cell as well as the local variables along the nuclear reactor operation. As a consequence, it is impractical to compute them for each possible configuration. An alternative to generate accurate pin-by-pin interface discontinuity factors is to calculate reference values using zero-net-current boundary conditions and to synthesize afterwards their dependencies on the main neighborhood variables. In such way the factors can be accurately computed during fine-mesh diffusion calculations by correcting the reference values as a function of the actual environment of the pin-cell in the core. In this paper we propose a parameterization of the pin-by-pin interface discontinuity factors allowing the implementation of a cross sections library able to treat the neighborhood effect. First results are presented for typical PWR configurations. (author)

Enquiry into the Topology of Plasma Membrane-Localized PIN Auxin Transport Components.

Science.gov (United States)

Nodzyński, Tomasz; Vanneste, Steffen; Zwiewka, Marta; Pernisová, Markéta; Hejátko, Jan; Friml, Jiří

2016-11-07

Auxin directs plant ontogenesis via differential accumulation within tissues depending largely on the activity of PIN proteins that mediate auxin efflux from cells and its directional cell-to-cell transport. Regardless of the developmental importance of PINs, the structure of these transporters is poorly characterized. Here, we present experimental data concerning protein topology of plasma membrane-localized PINs. Utilizing approaches based on pH-dependent quenching of fluorescent reporters combined with immunolocalization techniques, we mapped the membrane topology of PINs and further cross-validated our results using available topology modeling software. We delineated the topology of PIN1 with two transmembrane (TM) bundles of five α-helices linked by a large intracellular loop and a C-terminus positioned outside the cytoplasm. Using constraints derived from our experimental data, we also provide an updated position of helical regions generating a verisimilitude model of PIN1. Since the canonical long PINs show a high degree of conservation in TM domains and auxin transport capacity has been demonstrated for Arabidopsis representatives of this group, this empirically enhanced topological model of PIN1 will be an important starting point for further studies on PIN structure-function relationships. In addition, we have established protocols that can be used to probe the topology of other plasma membrane proteins in plants. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

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.

Internal fuel pin oxidizer

International Nuclear Information System (INIS)

Andrews, M.G.

1978-01-01

A nuclear fuel pin has positioned within it material which will decompose to release an oxidizing agent which will react with the cladding of the pin and form a protective oxide film on the internal surface of the cladding

Biosynthesis of collagen in the lung of the mouse after X-irradiation

International Nuclear Information System (INIS)

Walklin, C.M.; Law, M.P.

1986-01-01

Increases in the activities of both prolyl-4-hydroxylase (P-4-Hase) and protein disulphide isomerase (PDI) were observed as early as 1 month after 5-9 Gy. Maximal increases were observed at 6-7 months after 9 Gy and persisted up to 15 months after exposure. Increases after 5 and 7.5 Gy were more gradual but by 1 year after irradiation they had reached levels similar to those after 9 Gy. Collagen types were analysed at 2, 7.5 and 15 months. Results are shown for 7.5 and 15 months after 9 Gy. Although the total collagen content was increased, the ratio of collagen type I to III was normal. (UK)

Design and construction of a novel 1H/19F double-tuned coil system using PIN-diode switches at 9.4T.

Science.gov (United States)

Choi, Chang-Hoon; Hong, Suk-Min; Ha, YongHyun; Shah, N Jon

2017-06-01

A double-tuned 1 H/ 19 F coil using PIN-diode switches was developed and its performance evaluated. The is a key difference from the previous developments being that this design used a PIN-diode switch in series with an additionally inserted inductor in parallel to one of the capacitors on the loop. The probe was adjusted to 19 F when the reverse bias voltage was applied (PIN-diode OFF), whilst it was switched to 1 H when forward current was flowing (PIN-diode ON). S-parameters and Q-factors of single- and double-tuned coils were examined and compared with/without a phantom on the bench. Imaging experiments were carried out on a 9.4T preclinical scanner. All coils were tuned at resonance frequencies and matched well. It is shown that the Q-ratio and SNR of double-tuned coil at 19 F frequency are nearly as good as those of a single-tuned coil. Since the operating frequency was tuned to 19 F when the PIN-diodes were turned off, losses due to PIN-diodes were substantially lower resulting in the provision of excellent image quality of X-nuclei. Copyright © 2017 Elsevier Inc. All rights reserved.

Whole-Pin Furnace system: An experimental facility for studying irradiated fuel pin behavior under potential reactor accident conditions

International Nuclear Information System (INIS)

Liu, Y.Y.; Tsai, H.C.; Donahue, D.A.; Pushis, D.O.; Savoie, F.E.; Holland, J.W.; Wright, A.E.; August, C.; Bailey, J.L.; Patterson, D.R.

1990-05-01

The whole-pin furnace system is a new in-cell experimental facility constructed to investigate how irradiated fuel pins may fail under potential reactor accident conditions. Extensive checkouts have demonstrated excellent performance in remote operation, temperature control, pin breach detection, and fission gas handling. The system is currently being used in testing of EBIR-II-irradiated Integral Fast Reactor (IFR) metal fuel pins; future testing will include EBR-II-irradiated mixed-oxide fuel pins. 7 refs., 4 figs

In-house SIRAS phasing of the polyunsaturated fatty-acid isomerase from Propionibacterium acnes

International Nuclear Information System (INIS)

Liavonchanka, Alena; Hornung, Ellen; Feussner, Ivo; Rudolph, Markus

2006-01-01

Low iodide concentrations were sufficient to allow SAD and SIRAS phasing of cubic crystals of a novel fatty acid isomerase using Cu Kα radiation. The polyenoic fatty-acid isomerase from Propionibacterium acnes (PAI) catalyzes the double-bond isomerization of linoleic acid to conjugated linoleic acid, which is a dairy- or meat-derived fatty acid in the human diet. PAI was overproduced in Escherichia coli and purified to homogeneity as a yellow-coloured protein. The nature of the bound cofactor was analyzed by absorption and fluorescence spectroscopy. Single crystals of PAI were obtained in two crystal forms. Cubic shaped crystals belong to space group I2 1 3, with a unit-cell parameter of 160.4 Å, and plate-like crystals belong to the monoclinic space group C2, with unit-cell parameters a = 133.7, b = 60.8, c = 72.2 Å, β = 115.8°. Both crystal forms contain one molecule per asymmetric unit and diffract to a resolution of better than 2.0 Å. Initial phases were obtained by SIRAS from in-house data from a cubic crystal that was soaked with an unusually low KI concentration of 0.25 M

Cyclophilin B as a co-regulator of prolactin-induced gene expression and function in breast cancer cells

OpenAIRE

Fang, Feng; Zheng, Jiamao; Galbaugh, Traci L; Fiorillo, Alyson A; Hjort, Elizabeth E; Zeng, Xianke; Clevenger, Charles V

2010-01-01

The effects of prolactin (PRL) during the pathogenesis of breast cancer are mediated in part though Stat5 activity enhanced by its interaction with its transcriptional inducer, the prolyl isomerase cyclophilin B (CypB). We have demonstrated that knockdown of CypB decreases cell growth, proliferation, and migration, and CypB expression is associated with malignant progression of breast cancer. In this study, we examined the effect of CypB knockdown on PRL signaling in breast cancer cells. CypB...

Analytical method for reconstruction pin to pin of the nuclear power density distribution

International Nuclear Information System (INIS)

Pessoa, Paulo O.; Silva, Fernando C.; Martinez, Aquilino S.

2013-01-01

An accurate and efficient method for reconstructing pin to pin of the nuclear power density distribution, involving the analytical solution of the diffusion equation for two-dimensional neutron energy groups in homogeneous nodes, is presented. The boundary conditions used for analytic as solution are the four currents or fluxes on the surface of the node, which are obtained by Nodal Expansion Method (known as NEM) and four fluxes at the vertices of a node calculated using the finite difference method. The analytical solution found is the homogeneous distribution of neutron flux. Detailed distributions pin to pin inside a fuel assembly are estimated by the product of homogeneous flux distribution by local heterogeneous form function. Furthermore, the form functions of flux and power are used. The results obtained with this method have a good accuracy when compared with reference values. (author)

Analytical method for reconstruction pin to pin of the nuclear power density distribution

Energy Technology Data Exchange (ETDEWEB)

Pessoa, Paulo O.; Silva, Fernando C.; Martinez, Aquilino S., E-mail: ppessoa@con.ufrj.br, E-mail: fernando@con.ufrj.br, E-mail: aquilino@imp.ufrj.br [Coordenacao dos Programas de Pos-Graduacao em Engenharia (COPPE/UFRJ), Rio de Janeiro, RJ (Brazil)

2013-07-01

An accurate and efficient method for reconstructing pin to pin of the nuclear power density distribution, involving the analytical solution of the diffusion equation for two-dimensional neutron energy groups in homogeneous nodes, is presented. The boundary conditions used for analytic as solution are the four currents or fluxes on the surface of the node, which are obtained by Nodal Expansion Method (known as NEM) and four fluxes at the vertices of a node calculated using the finite difference method. The analytical solution found is the homogeneous distribution of neutron flux. Detailed distributions pin to pin inside a fuel assembly are estimated by the product of homogeneous flux distribution by local heterogeneous form function. Furthermore, the form functions of flux and power are used. The results obtained with this method have a good accuracy when compared with reference values. (author)

Characterization of an L-arabinose isomerase from Bacillus thermoglucosidasius for D-tagatose production.

Science.gov (United States)

Seo, Myung-Ji

2013-01-01

L-Arabinose isomerase from Bacillus thermoglucosidasius KCTC 1828 (BTAI) was expressed in Escherichia coli. The optimal temperature and pH for the activity of the purified BTAI were 40 °C and pH 7.0. The Mn(2+) ion was an activator of BTAI activity. The kinetic parameters of BTAI for D-galactose were a K(m) of 175 mM and a k(cat)/K(m) of 2.8 mM(-1)min(-1). The conversion ratio by BTAI to D-tagatose reached 45.6% at 40 °C.

Optimization study on pin tip diameter of an impact-pin nozzle at high pressure ratio

Energy Technology Data Exchange (ETDEWEB)

Kumar, C. Palani; Lee, Kwon Hee [FMTRC, Daejoo Machinery Co. Ltd., Daegu (Korea, Republic of); Park, Tae Choon; Cha, Bong Jun [Engine Components Research Team, Korea Aerospace Research Institute, Daejeon (Korea, Republic of); Kim, Heuy Dong [Dept. of Mechanical Engineering, Andong National University, Andong (Korea, Republic of)

2016-09-15

Wet compression system is typically installed in a gas turbine engine to increase the net power output and efficiency. A crucial component of the wet compression system is the nozzle which generates fine water droplets for injection into the compressor. The main objective of present work is to optimize a kind of nozzle called impact-pin spray nozzle and thereby produce better quality droplets. To achieve this, the dynamics occurring in the water jet impinging on the pin tip, the subsequent formation of water sheet, which finally breaks into water droplets, must be studied. In this manuscript, the progress on the numerical studies on impact-pin nozzle are reported. A small computational domain covering the orifice, pin tip and the region where primary atomization occurs is selected for numerical analysis. The governing equations are selected in three dimensional cartesian form and simulations are performed to predict the dynamics of water jet impinging on the pin. Systematic studies were carried out and the results leading to the choice of turbulence model and the effect of pin tip diameter are reported here. Further studies are proposed to show the future directions of the present research work.

Self-organized critical behavior in pinned flux lattices

International Nuclear Information System (INIS)

Pla, O.; Nori, F.

1991-01-01

We study the response of pinned fluxed lattices, under small perturbations in the driving force, below and close to the pinning-depinning transition. For driving Lorentz forces below F c (the depinning force at which the whole flux lattice slides), the system has instabilities against small force increases, with a power-law distribution characteristic of self-organized criticality. Specifically, D(d)∼d -1,3 , where d is the displacement of a flux line after a very small force increase. We also study the initial stages of the motion of the lattice once the driving force overcomes the pinning forces

Top-nozzle mounted replacement guide pin assemblies

International Nuclear Information System (INIS)

Gilmore, C.B.; Andrews, W.H.

1993-01-01

A replacement guide pin assembly is provided for aligning a nuclear fuel assembly with an upper core plate of a nuclear reactor core. The guide pin assembly includes a guide pin body having a radially expandable base insertable within a hole in the top nozzle, a ferrule insertable within the guide pin base and capable of imparting a radially and outwardly directed force on the expandable base to expand it within the hole of the top nozzle and thereby secure the guide pin body to the top nozzle in response to a predetermined displacement of the ferrule relative to the guide pin body along its longitudinal axis, and a lock screw interfitted with the ferrule and threaded into the guide pin body so as to produce the predetermined displacement of the ferrule. (author)

Fuel pin bowing and related investigation of WWER-440 control rod influence on power release inside of neighbouring fuel pins

International Nuclear Information System (INIS)

Mikus, J.

2005-01-01

The purpose of this work consists in investigation of the WWER-440 control rod (CR) influence on space power distribution, especially from viewpoint of the values and gradient occurrence that could result in static and cyclic loads with some consequences, e.g. fuel pin bowing. As known, CR can cause power peaks in periphery fuel pins of adjacent operating assemblies because of the butt joint design of the absorbing adapter to the CR fuel part, that is, presence of the water cavity resulting in a flash up of thermal neutrons. As a consequence, beside well-known peaks in axial power distribution, above power gradients can occur inside of mentioned fuel pins. Because of complicated geometry and material composition of the CR, the detailed calculations concerning both above phenomena are complicated, too. Therefore it is useful to acquire appropriate experimental data to investigate mentioned influence and compare them with calculations. Since detailed power distributions cannot be obtained in the NPP, needed information is provided by means of experiments on research reactors. In case of measurements inside of fuel pins, special (e.g. track) detectors placed between fuel pellets are used. Such works are relatively complicated and time consuming, therefore an evaluation based on mathematical modelling and numerical approximation was proposed by means of that, and using measured power release in some selected fuel pins, information about power release inside of one of these fuel pins, can be obtained. For this purpose, an experiment on light water, zero-power research reactor LR-0 was realized and axial power distribution measurements were performed in a WWER-440 type core near to an authentic CR model. Application of the above evaluation method is demonstrated on one ''investigated'' fuel pin neighbouring CR by means of following results: 1. Axial power distribution inside of investigated fuel pin in two opposite positions on its pellets surface that are situated to

Styrene Oxide Isomerase of Rhodococcus opacus 1CP, a Highly Stable and Considerably Active Enzyme

Science.gov (United States)

Gröning, Janosch A. D.; Tischler, Dirk; Kaschabek, Stefan R.; Schlömann, Michael

2012-01-01

Styrene oxide isomerase (SOI) is involved in peripheral styrene catabolism of bacteria and converts styrene oxide to phenylacetaldehyde. Here, we report on the identification, enrichment, and biochemical characterization of a novel representative from the actinobacterium Rhodococcus opacus 1CP. The enzyme, which is strongly induced during growth on styrene, was shown to be membrane integrated, and a convenient procedure was developed to highly enrich the protein in active form from the wild-type host. A specific activity of about 370 U mg−1 represents the highest activity reported for this enzyme class so far. This, in combination with a wide pH and temperature tolerance, the independence from cofactors, and the ability to convert a spectrum of substituted styrene oxides, makes a biocatalytic application imaginable. First, semipreparative conversions were performed from which up to 760 μmol of the pure phenylacetaldehyde could be obtained from 130 U of enriched SOI. Product concentrations of up to 76 mM were achieved. However, due to the high chemical reactivity of the aldehyde function, SOI was shown to be the subject of an irreversible product inhibition. A half-life of 15 min was determined at a phenylacetaldehyde concentration of about 55 mM, indicating substantial limitations of applicability and the need to modify the process. PMID:22504818

SAXS-WAXS studies of the low-resolution structure in solution of xylose/glucose isomerase from Streptomyces rubiginosus

Science.gov (United States)

Kozak, Maciej; Taube, Michał

2009-10-01

The structure and conformation of molecule of xylose/glucose isomerase from Streptomyces rubiginosus in solution (at pH 6 and 7.6; with and without the substrate) has been studied by small- and wide-angle scattering of synchrotron radiation (SAXS-WAXS). On the basis of the SAXS-WAXS data, the low-resolution structure in solution has been reconstructed using ab inito methods. A comparison of the models of glucose isomerase shows only small differences between the model in solution and the crystal structure.

Mode of failure of LMFBR fuel pins

International Nuclear Information System (INIS)

Washburn, D.F.

1975-01-01

The objectives of the irradiation test described were to evaluate mixed-oxide fuel performance and to confirm the design adequacy of the FFTF fuel pins. After attainment of the initial objectives the irradiation of several of the original fuel pins was continued until a cladding breach occurred. The consequences of a cladding breach were evaluated by reconstituting the original 37-pin subassembly into two 19-pin subassemblies after a burnup at 50,000 MWd/MTM (5.2 a/o). The original pins were supplemented with fresh pins as necessary. Irradiation of the subassemblies was continued until a cladding breach occurred. Results are presented and discussed

75 MeV boron ion irradiation studies on Si PIN photodiodes

Energy Technology Data Exchange (ETDEWEB)

Prabhakara Rao, Y.P.; Praveen, K.C. [Department of Studies in Physics, University of Mysore, Manasagangotri, Mysore 570006 (India); Rejeena Rani, Y. [Integrated Circuits Division, Bharat Electronics Limited, Bangalore 560013, Karnataka (India); Tripathi, Ambuj [Inter University Accelerator Centre, Aruna Asaf Ali Marg, New Delhi 110067 (India); Gnana Prakash, A.P., E-mail: gnanap@hotmail.com [Department of Studies in Physics, University of Mysore, Manasagangotri, Mysore 570006 (India)

2013-12-01

The highly sensitive silicon PIN photodiodes were fabricated to use in radiation environments. The Si PIN photodiodes are coated with 150 nm silicon dioxide (SiO{sub 2}) as anti-reflective (AR) coating. The presence of AR coating on the performance of irradiated PIN photodiodes is studied up to a total dose of 10 Mrad. The effects of 75 MeV boron (B{sup 5+}) ions and {sup 60}Co gamma radiation on the I–V, C–V and spectral responses of PIN photodiodes were studied systematically to understand the radiation tolerance of the devices. The 75 MeV B{sup 5+} irradiation results are compared with {sup 60}Co gamma irradiated results in the same dose range for 1 mm × 1 mm and 10 mm × 10 mm active area PIN photodiodes. The irradiation results show that the ion irradiated PIN photodiodes show more degradation when compared {sup 60}Co gamma irradiated devices. The irradiation results are presented in this paper and the possible mechanism behind the degradation of photodiodes is also discussed in the paper.

Flux pinning and flux flow studies in superconductors using flux flow noise techniques. Progress report, April 1, 1976--December 17, 1976

International Nuclear Information System (INIS)

Joiner, W.C.H.

1976-12-01

Measurements of flux flow noise power spectra have been combined with critical current measurements and measurements of current-voltage characteristics to study flux flow and local pinning interactions effective during flux flow. A model of flux flow noise generation in the presence of local pinning interactions is developed and applied to situations where pinning is dominated by: (1) grain boundaries, (2) normal metal precipitates in a superconducting matrix, (3) gross deformation producing a critical current peak effect, and (4) surface grooves imposed on a sample surface. In the case of pinning caused by normal metal precipitates in a superconducting matrix, unusual training and hysterisis effects are observed in the flux flow characteristics. The greater sensitivity of noise spectra, as compared with bulk critical current measurements, in obtaining a detailed picture of flux flow is emphasized

BARS - a heterogeneous code for 3D pin-by-pin LWR steady-state and transient calculation

International Nuclear Information System (INIS)

Avvakumov, A.V.; Malofeev, V.M.

2000-01-01

A 3D pin-by-pin dynamic model for LWR detailed calculation was developed. The model is based on a coupling of the BARS neutronic code with the RELAP5/MOD3.2 thermal hydraulic code. This model is intended to calculate a fuel cycle, a xenon transient, and a wide range of reactivity initiated accidents in a WWER and a PWR. Galanin-Feinberg heterogeneous method was realized in the BARS code. Some results for a validation of the heterogeneous method are presented for reactivity coefficients, a pin-by-pin power distribution, and a fast pulse transient. (Authors)

Temperature and pinning strength dependence of the critical current of a superconductor with a square periodic array of pinning sites

International Nuclear Information System (INIS)

Benkraouda, M.; Obaidat, I.M.; Al Khawaja, U.

2006-01-01

We have conducted extensive series of molecular dynamic simulations on driven vortex lattices interacting with periodic square arrays of pinning sites. In solving the over damped equation of vortex motion we took into account the vortex-vortex repulsion interaction, the attractive vortex-pinning interaction, and the driving Lorentz force at several values of temperature. We have studied the effect of varying the driving Lorentz force and varying the pinning strength on the critical current for several pinning densities, and temperature values. We have found that the pinning strength play an important role in enhancing the critical current over the whole temperature range. At low temperatures, the critical current was found to increase linearly with increasing the pinning strengths for all pinning densities. As the temperature increases, the effect of small pinning strengths diminishes and becomes insignificant at high temperatures

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...

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.

Development of spectral history methods for pin-by-pin core analysis method using three-dimensional direct response matrix

International Nuclear Information System (INIS)

Mitsuyasu, T.; Ishii, K.; Hino, T.; Aoyama, M.

2009-01-01

Spectral history methods for pin-by-pin core analysis method using the three-dimensional direct response matrix have been developed. The direct response matrix is formalized by four sub-response matrices in order to respond to a core eigenvalue k and thus can be recomposed at each outer iteration in the core analysis. For core analysis, it is necessary to take into account the burn-up effect related to spectral history. One of the methods is to evaluate the nodal burn-up spectrum obtained using the out-going neutron current. The other is to correct the fuel rod neutron production rates obtained the pin-by-pin correction. These spectral history methods were tested in a heterogeneous system. The test results show that the neutron multiplication factor error can be reduced by half during burn-up, the nodal neutron production rates errors can be reduced by 30% or more. The root-mean-square differences between the relative fuel rod neutron production rate distributions can be reduced within 1.1% error. This means that these methods can accurately reflect the effects of intra- and inter-assembly heterogeneities during burn-up and can be used for core analysis. Core analysis with the DRM method was carried out for an ABWR quarter core and it was found that both thermal power and coolant-flow distributions were smoothly converged. (authors)

Pinning synchronization of a mobile agent network

International Nuclear Information System (INIS)

Wang, Lei; Sun, You-xian

2009-01-01

We investigate the problem of controlling a group of mobile agents in a plane in order to move them towards a desired orbit via pinning control, in which each agent is associated with a chaotic oscillator coupled with those of neighboring agents, and the pinning strategy is to have the common linear feedback acting on a small fraction of agents by random selection. We explore the effects of the pinning probability, feedback gains and agent density in the pinning synchronization of a mobile agent network under a fast-switching constraint, and perform numerical simulations for validation. In particular, we show that there exists a critical pinning density for network synchronization with an unbounded region: above the threshold, the dynamical network can be controlled by pinning; below it, anarchy prevails. And for the network with a single bounded synchronization region, pinning control has little effect as regards enhancing network synchronizability

Protein disulfide isomerase-like protein 1-1 controls endosperm development through regulation of the amount and composition of seed proteins in rice.

Directory of Open Access Journals (Sweden)

Yeon Jeong Kim

Full Text Available Protein disulfide isomerase (PDI is a chaperone protein involved in oxidative protein folding by acting as a catalyst and assisting folding in the endoplasmic reticulum (ER. A genome database search showed that rice contains 19 PDI-like genes. However, their functions are not clearly identified. This paper shows possible functions of rice PDI-like protein 1-1 (PDIL1-1 during seed development. Seeds of the T-DNA insertion PDIL1-1 mutant, PDIL1-1Δ, identified by genomic DNA PCR and western blot analysis, display a chalky phenotype and a thick aleurone layer. Protein content per seed was significantly lower and free sugar content higher in PDIL1-1Δ mutant seeds than in the wild type. Proteomic analysis of PDIL1-1Δ mutant seeds showed that PDIL1-1 is post-translationally regulated, and its loss causes accumulation of many types of seed proteins including glucose/starch metabolism- and ROS (reactive oxygen species scavenging-related proteins. In addition, PDIL1-1 strongly interacts with the cysteine protease OsCP1. Our data indicate that the opaque phenotype of PDIL1-1Δ mutant seeds results from production of irregular starch granules and protein body through loss of regulatory activity for various proteins involved in the synthesis of seed components.

Vortex lattice melting, pinning and kinetics

International Nuclear Information System (INIS)

Doniach, S.; Ryu, S.; Kapitulnik, A.

1994-01-01

The phenomenology of the high T c superconductors is discussed both at the level of the thermodynamics of melting of the Abrikosov flux lattice and in terms of the melting and kinetics of the flux lattice for a pinned system. The authors review results on 3D melting obtained by a Monte Carlo simulation approach in which the 2D open-quotes pancakeclose quotes vortices are treated as statistical variables. The authors discuss pinning in the context of the strong pinning regime in which the vortex density given in terms of the applied field B is small compared to that represented by an effective field B pin measuring the pinning center density. The authors introduce a new criterion for the unfreezing of a vortex glass on increase of magnetic field or temperature, in the strong pinning, small field unit. The authors model this limit in terms of a single flux line interacting with a columnar pin. This model is studied both analytically and by computer simulation. By applying a tilt potential, the authors study the kinetics of the vortex motion in an external current and show that the resulting current-voltage characteristic follows a basic vortex glass-like scaling relation in the vicinity of the depinning transition

Pin fin compliant heat sink with enhanced flexibility

Science.gov (United States)

Schultz, Mark D.

2018-04-10

Heat sinks and methods of using the same include a top and bottom plate, at least one of which has a plurality of pin contacts flexibly connected to one another, where the plurality of pin contacts have vertical and lateral flexibility with respect to one another; and pin slice layers, each having multiple pin slices, arranged vertically between the top and bottom plates such that the plurality of pin slices form substantially vertical pins connecting the top and bottom plates.

Expression of Cyclophilin B is Associated with Malignant Progression and Regulation of Genes Implicated in the Pathogenesis of Breast Cancer

OpenAIRE

Fang, Feng; Flegler, Ayanna J.; Du, Pan; Lin, Simon; Clevenger, Charles V.

2009-01-01

Cyclophilin B (CypB) is a 21-kDa protein with peptidyl-prolyl cis-trans isomerase activity that functions as a transcriptional inducer for Stat5 and as a ligand for CD147. To better understand the global function of CypB in breast cancer, T47D cells with a small interfering RNA-mediated knockdown of CypB were generated. Subsequent expression profiling analysis showed that 663 transcripts were regulated by CypB knockdown, and that many of these gene products contributed to cell proliferation, ...

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

Multilevel parallel strategy on Monte Carlo particle transport for the large-scale full-core pin-by-pin simulations

International Nuclear Information System (INIS)

Zhang, B.; Li, G.; Wang, W.; Shangguan, D.; Deng, L.

2015-01-01

This paper introduces the Strategy of multilevel hybrid parallelism of JCOGIN Infrastructure on Monte Carlo Particle Transport for the large-scale full-core pin-by-pin simulations. The particle parallelism, domain decomposition parallelism and MPI/OpenMP parallelism are designed and implemented. By the testing, JMCT presents the parallel scalability of JCOGIN, which reaches the parallel efficiency 80% on 120,000 cores for the pin-by-pin computation of the BEAVRS benchmark. (author)

Development of 3D pseudo pin-by-pin calculation methodology in ANC

International Nuclear Information System (INIS)

Zhang, B.; Mayhue, L.; Huria, H.; Ivanov, B.

2012-01-01

Advanced cores and fuel assembly designs have been developed to improve operational flexibility, economic performance and further enhance safety features of nuclear power plants. The simulation of these new designs, along with strong heterogeneous fuel loading, have brought new challenges to the reactor physics methodologies currently employed in the industrial codes for core analyses. Control rod insertion during normal operation is one operational feature in the AP1000 R plant of Westinghouse next generation Pressurized Water Reactor (PWR) design. This design improves its operational flexibility and efficiency but significantly challenges the conventional reactor physics methods, especially in pin power calculations. The mixture loading of fuel assemblies with significant neutron spectrums causes a strong interaction between different fuel assembly types that is not fully captured with the current core design codes. To overcome the weaknesses of the conventional methods, Westinghouse has developed a state-of-the-art 3D Pin-by-Pin Calculation Methodology (P3C) and successfully implemented in the Westinghouse core design code ANC. The new methodology has been qualified and licensed for pin power prediction. The 3D P3C methodology along with its application and validation will be discussed in the paper. (authors)

Investigation of gene expression and serum levels of PIN1 and eNOS with high blood pressure in patients with Alzheimer disease.

Science.gov (United States)

Azimi, Mina; Nikanfar, Masoud; Khakikhatibi, Fatemeh; Rahbarghazi, Reza; Nourazarian, Seyed Manuchehr; Biray Avci, Cigir; Nourazarian, Alireza

2017-09-01

According to evidence, Alzheimer's disease is known as one of the most serious neurodegenerative diseases, for which hypertension has been observed to be a key risk factor. Therefore, this study aims to examine the relationship between the PIN1 and eNOS genes expression, as well as serum levels and hypertension in Alzheimer's disease sufferers. Blood samples were obtained from subjects who were divided into four groups: the control group, normotensive Alzheimer's patients, the Alzheimer's sufferers group with hypertension, and the healthy group with only hypertension, considering the inhibition of confounding factors. Thereafter, eNOS and PIN1 genes expression along with serum levels were studied. Based on the obtained results, a statistically significant correlation didn't exist between serum level of PIN1 and the systolic and diastolic blood pressure, between serum level of eNOS and diastolic blood pressure in the norm tension Alzheimer's disease patients, between serum levels of PIN1, eNOS and systolic blood pressure, and between serum eNOS and systolic and diastolic blood pressure in the patients with hypertension (pdiseases in Alzheimer's disease. Copyright © 2017 Elsevier Ltd. All rights reserved.

Performance of fast reactor mixed-oxide fuels pins during extended overpower transients

International Nuclear Information System (INIS)

Tsai, H.; Neimark, L.A.; Asaga, T.; Shikakura, S.

1991-02-01

The Operational Reliability Testing (ORT) program, a collaborative effort between the US Department of Energy and the Power Reactor and Nuclear Fuel Development Corp. (PNC) of Japan, was initiated in 1982 to investigate the behavior of mixed-oxide fuel pin under various slow-ramp transient and duty-cycle conditions. In the first phase of the program, a series of four extended overpower transient tests, with severity sufficient to challenge the pin cladding integrity, was conducted. The objectives of the designated TOPI-1A through -1D tests were to establish the cladding breaching threshold and mechanisms, and investigate the thermal and mechanical effects of the transient on pin behavior. The tests were conducted in EBR-2, a normally steady-state reactor. The modes of transient operation in EBR-2 were described in a previous paper. Two ramp rates, 0.1%/s and 10%/s, were selected to provide a comparison of ramp-rate effects on fuel behavior. The test pins chosen for the series covered a range of design and pre-test irradiation parameters. In the first test (1A), all pins maintained their cladding integrity during the 0.1%/s ramp to 60% peak overpower. Fuel pins with aggressive designs, i.e., high fuel- smear density and/or thin cladding, were, therefore, included in the follow-up 1B and 1C tests to enhance the likelihood of achieving cladding breaching. In the meantime, a higher pin overpower capability, to greater than 100%, was established by increasing the reactor power limit from 62.5 to 75 MWt. In this paper, the significant results of the 1B and 1C tests are presented. 4 refs., 5 figs., 1 tab

Timing analysis of PWR fuel pin failures

International Nuclear Information System (INIS)

Jones, K.R.; Wade, N.L.; Katsma, K.R.; Siefken, L.J.; Straka, M.

1992-09-01

Research has been conducted to develop and demonstrate a methodology for calculation of the time interval between receipt of the containment isolation signals and the first fuel pin failure for loss-of-coolant accidents (LOCAs). Demonstration calculations were performed for a Babcock and Wilcox (B ampersand W) design (Oconee) and a Westinghouse (W) four-loop design (Seabrook). Sensitivity studies were performed to assess the impacts of fuel pin bumup, axial peaking factor, break size, emergency core cooling system availability, and main coolant pump trip on these times. The analysis was performed using the following codes: FRAPCON-2, for the calculation of steady-state fuel behavior; SCDAP/RELAP5/MOD3 and TRACPF1/MOD1, for the calculation of the transient thermal-hydraulic conditions in the reactor system; and FRAP-T6, for the calculation of transient fuel behavior. In addition to the calculation of fuel pin failure timing, this analysis provides a comparison of the predicted results of SCDAP/RELAP5/MOD3 and TRAC-PFL/MOD1 for large-break LOCA analysis. Using SCDAP/RELAP5/MOD3 thermal-hydraulic data, the shortest time intervals calculated between initiation of containment isolation and fuel pin failure are 10.4 seconds and 19.1 seconds for the B ampersand W and W plants, respectively. Using data generated by TRAC-PF1/MOD1, the shortest intervals are 10.3 seconds and 29.1 seconds for the B ampersand W and W plants, respectively. These intervals are for a double-ended, offset-shear, cold leg break, using the technical specification maximum peaking factor and applied to fuel with maximum design bumup. Using peaking factors commensurate widi actual bumups would result in longer intervals for both reactor designs. This document also contains appendices A through J of this report

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.

Integral Fast Reactor fuel pin processor

International Nuclear Information System (INIS)

Levinskas, D.

1993-01-01

This report discusses the pin processor which receives metal alloy pins cast from recycled Integral Fast Reactor (IFR) fuel and prepares them for assembly into new IFR fuel elements. Either full length as-cast or precut pins are fed to the machine from a magazine, cut if necessary, and measured for length, weight, diameter and deviation from straightness. Accepted pins are loaded into cladding jackets located in a magazine, while rejects and cutting scraps are separated into trays. The magazines, trays, and the individual modules that perform the different machine functions are assembled and removed using remote manipulators and master-slaves

Effect of oil around the pin boss bearing on bearing friction force; Piston pin jikuukebu kinbo no junkatsuyu ga masatsuryoku ni oyobosu eikyo

Energy Technology Data Exchange (ETDEWEB)

Takiguchi, M; Suhara, T; Ato, S; Someya, T [Musashi Institute of Technology, Tokyo (Japan)

1997-10-01

Piston pin bearings are engine parts placed under severe lubricating condition because of the high unit load and temperature and the low sliding speeds. Therefore, they are vulnerable to many lubrication problems such as abnormal wear, scuffs and seizures. In our recent study, the bearing friction were measured using a original measuring device and it was found out that the lubricating conditions of piston pin boss bearings are non-fluid lubrication due to the oil starvation. In this study, we have also measured the lubricating oil behavior around the pin boss bearing using a special cylinder with glass window, and analyzed the relationship between the friction force and the oil behavior at the pin boss bearings in a actual operating gasoline engine. 2 refs., 9 figs., 1 tab.

Flux pinning in MOD YBCO films by chemical doping

International Nuclear Information System (INIS)

Zhou, Y X; Ghalsasi, S; Rusakova, I; Salama, K

2007-01-01

A novel nanomaterial synthesis technique has been developed to introduce 0D (particles), 1D (columnar defects) and 3D (domains) nanoscale pinning centres in MOD Y 1 Ba 2 Cu 3 O 7 (YBCO) coated conductors. We have succeeded in introducing nanoscale Y enriched particles, nanoscale 90 0 rotated Y 1/3 Sm 2/3 Ba 2 Cu 3 O 7 domains and nanoscale Zr enriched columnar defects into YBCO layers by different chemical doping. The pinning force density in Y 2 O 3 -doped YBCO film is found to be larger than that of pure YBCO film at all fields. Also it was found that YBCO films with Sm substituting for Y have yielded improved critical current density characteristics over a wide range of magnetic fields. Maximum pinning force densities exceeding 7 and 8 GN m -3 are obtained in 5% BZO-doped and Sm substituted YBCO films, respectively. Additionally, TEM studies revealed nanoscale Zr enriched columnar defects distributing in the matrix of the c-oriented YBCO film throughout the whole cross section. This indicates that chemical doping is a promising fabrication technique to create specific pinning landscapes in YBCO coated conductors

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

Science.gov (United States)

Daar, I O; Artymiuk, P J; Phillips, D C; Maquat, L E

1986-10-01

Triose-phosphate isomerase (TPI; D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1) deficiency is a recessive disorder that results in hemolytic anemia and neuromuscular dysfunction. To determine the molecular basis of this disorder, a TPI allele from two unrelated patients homozygous for TPI deficiency was compared with an allele from a normal individual. Each disease-associated sequence harbors a G X C----C X G transversion in the codon for amino acid-104 and specifies a structurally altered protein in which a glutamate residue is replaced by an aspartate residue. The importance of glutamate-104 to enzyme structure and function is implicated by its conservation in the TPI protein of all species that have been characterized to date. The glutamate-to-aspartate substitution results in a thermolabile enzyme as demonstrated by assays of TPI activity in cultured fibroblasts of each patient and cultured Chinese hamster ovary (CHO) cells that were stably transformed with the mutant alleles. Although this substitution conserves the overall charge of amino acid-104, the x-ray crystal structure of chicken TPI indicates that the loss of a side-chain methylene group (-CH2CH2COO- ---- -CH2COO-) is sufficient to disrupt the counterbalancing of charges that normally exists within a hydrophobic pocket of the native enzyme.

FABRICE process for the refrabrication of experimental pins in a hot cell, from pins pre-irradiated in power reactors

International Nuclear Information System (INIS)

Vignesoult, N.; Atabek, R.; Ducas, S.

1982-06-01

The Fabrice ''hot cell refabrication'' process for small pins from very long irradiated fuel elements was developed at the CEA to allow parametric studies of the irradiation behavior of pins from nuclear power plants. Since this operation required complete assurance of the validity of the process, qualification of the fabrication was performed on test pins, refabricated in the hot cell, as well as irradiation qualification. The latter qualification was intended to demonstrate that, in identical experimental irradiation conditions, the refabricated Fabrice pins behaved in the same way as whole pins with the same initial characteristics. This qualification of the Fabrice process, dealing with more than twenty pins at different burnups, showed that fabrication did not alter: the inherent characteristics of the sampled fuel element and the irradiation behavior of the sampled fuel element [fr

A pinning puzzle: two similar, non-superconducting chemical deposits in YBCO-one pins, the other does not

Energy Technology Data Exchange (ETDEWEB)

Sawh, Ravi-Persad; Weinstein, Roy; Gandini, Alberto; Skorpenske, Harley; Parks, Drew, E-mail: Weinstein@uh.ed [Beam Particle Dynamics Laboratories, University of Houston, Houston, TX 77204-5005 (United States); Department of Physics, University of Houston, Houston, TX 77204-5005 (United States); Texas Center for Superconductivity at UH, University of Houston, Houston, TX 77204-5002 (United States)

2009-09-15

The pinning effects of two kinds of U-rich deposits in YBCO (YBa{sub 2}Cu{sub 3}O{sub 7-{delta}}) are compared. One is a five-element compound, (U{sub 0.6}Pt{sub 0.4})YBa{sub 2}O{sub 6}, which is a paramagnetic double perovskite which forms as profuse stable nanosize deposits, and pins very well. The other is a four-element compound, (U{sub 0.4}Y{sub 0.6})BaO{sub 3}, which is a ferromagnetic single perovskite which forms as profuse stable nanosize deposits and pins very weakly or not at all. The pinning comparison is done with nearly equal deposit sizes and number of deposits per unit volume for the two compounds. Evidence for the pinning capability, chemical makeup, x-ray diffraction signature, and magnetic properties of the two compounds is reported.

Tapered leaf support pin for operating plant guide tubes

International Nuclear Information System (INIS)

Land, J.T.; Hopkins, R.J.; Ford, D.E.

1991-01-01

This patent describes a mounting system for removably mounting the lower flange of a control rod guide tube over an opening in the upper core plate of a nuclear reactor comprising at least one elongated support pin mounted on the guide tube lower flange and resiliently receivable in a bore formed in the upper core plate. It comprises a support pin having a longitudinal axis and comprising a first pin portion mountable on the guide tube lower flange, and a second pin portion receivable within the upper core plate bore, the second pin portion including a solid body section adjacent the first pin portion and having an outer diameter which is accommodated by the bore by a close clearance fit; locking means mounted on the first pin portion of the support pin for retaining the guide tube lower flange between the solid body section of the second pin portion and the locking means; and a washer disposed around the first pin portion between the locking means and the control rod guide tube flange, the washer and the locking means including mutually engaging rounded surfaces for eliminating bending moments and stresses on the support pin during mounting of the locking means on the first pin portion of the support pin

Irradiation of TZM: Uranium dioxide fuel pin at 1700 K

Science.gov (United States)

Mcdonald, G. E.

1973-01-01

A fuel pin clad with TZM and containing solid pellets of uranium dioxide was fission heated in a static helium-cooled capsule at a maximum surface temperature of 1700 K for approximately 1000 hr and to a total burnup of 2.0 percent of the uranium-235. The results of the postirradiation examination indicated: (1) A transverse, intergranular failure of the fuel pin occurred when the fuel pin reached 2.0-percent burnup. This corresponds to 1330 kW-hr/cu cm, where the volume is the sum of the fuel, clad, and void volumes in the fuel region. (2) The maximum swelling of the fuel pin was less than 1.5 percent on the fuel-pin diameter. (3) There was no visible interaction between the TZM clad and the UO2. (4) Irradiation at 1700 K produced a course-grained structure, with an average grain diameter of 0.02 centimeter and with some of the grains extending one-half of the thickness of the clad. (5) Below approximately 1500 K, the irradiation of the clad produced a moderately fine-grained structure, with an average grain diameter of 0.004 centimeter.

Direct production of D-arabinose from D-xylose by a coupling reaction using D-xylose isomerase, D-tagatose 3-epimerase and D-arabinose isomerase.

Science.gov (United States)

Sultana, Ishrat; Mizanur, Rahman Md; Takeshita, Kei; Takada, Goro; Izumori, Ken

2003-01-01

Klebsiella pneumoniae 40bXX, a mutant strain that constitutively produces D-arabinose isomerase (D-AI), was isolated through a series of repeated subcultures from the parent strain on a mineral salt medium supplemented with L-Xylose as the sole carbon source. D-AI could be efficiently immobilized on chitopearl beads. The optimum temperature for the activity of the immobilized enzyme was 40 degrees C and the enzyme was stable up to 50 degrees C. The D-Al was active at pH 10.0 and was stable in the range of pH 6.0-11.0. The enzyme required manganese ions for maximum activity. Three immobilized enzymes, D-xylose isomerase (D-XI), D-tagatose 3-epimerase (D-TE and D-AI were used for the preparation of D-arabinose from D-xylose in a coupling reaction. After completion of the reaction, degradation of D-xylulose was carried out by Saccharomyces cerevisiae. The reaction mixture containing D-Xylose, D-ribulose and the product was then separated by ion exchange column chromatography. After crystallization, the product was checked by HPLC, IR spectroscopy, NMR spectroscopy and optical rotation measurements. Finally, 2.0 g of D-arabinose could be obtained from 5 g of the substrate.

Cyclophilin B stimulates RNA synthesis by the HCV RNA dependent RNA polymerase.

Science.gov (United States)

Heck, Julie A; Meng, Xiao; Frick, David N

2009-04-01

Cyclophilins are cellular peptidyl isomerases that have been implicated in regulating hepatitis C virus (HCV) replication. Cyclophilin B (CypB) is a target of cyclosporin A (CsA), an immunosuppressive drug recently shown to suppress HCV replication in cell culture. Watashi et al. recently demonstrated that CypB is important for efficient HCV replication, and proposed that it mediates the anti-HCV effects of CsA through an interaction with NS5B [Watashi K, Ishii N, Hijikata M, Inoue D, Murata T, Miyanari Y, et al. Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase. Mol Cell 2005;19:111-22]. We examined the effects of purified CypB proteins on the enzymatic activity of NS5B. Recombinant CypB purified from insect cells directly stimulated NS5B-catalyzed RNA synthesis. CypB increased RNA synthesis by NS5B derived from genotype 1a, 1b, and 2a HCV strains. Stimulation appears to arise from an increase in productive RNA binding. NS5B residue Pro540, a previously proposed target of CypB peptidyl-prolyl isomerase activity, is not required for stimulation of RNA synthesis.

HEPATOTROPHIC EFFECTS OF FK506 IN DOGS1

Science.gov (United States)

Starzl, Thomas E.; Porter, Kendrick A.; Mazzaferro, Vincenzo; Todo, Satoru; Fung, John; Francavilla, Antonio

2010-01-01

Portacaval shunt (Eck fistula) in dogs causes hepatocyte atrophy and organelle disruption, as well as tripling of hepatocyte mitoses. After submitting dogs to this procedure, FK506 was infused into the tied-off left portal vein. The size, anatomic quality, and replication of hepatocytes were enhanced in the portion of liver infused with FK506, with a significant spillover effect in the noninfused portion. These hepatotrophic qualities of FK506 may explain part of FK506’s efficacy for the treatment of chronic liver rejection. Also, the observations support a trial with this drug for the treatment of autoimmune liver diseases because, in addition to turning off the immunologic genesis of such disorders, repair and regeneration of the damaged liver may be augmented. Finally, these hepatrophic qualities are part of an emerging spectrum of biologic effects caused by drugs that may modulate the enzyme cis-trans peptidyl-prolyl isomerase (PPIase), the principal constituent of the cytosolic binding sites of FK506, repamycin, cyclosporine, and presumably other immunosuppressive drugs as yet undiscovered. PMID:1702912

Performance of IN-706 and PE-16 cladding in mixed-oxide fuel pins

International Nuclear Information System (INIS)

Makenas, B.J.; Lawrence, L.A.; Jensen, B.W.

1982-05-01

Iron-nickel base, precipitation-strengthened alloys, IN-706 and PE-16, advanced alloy cladding considered for breeder reactor applications, were irradiated in mixed-oxide fuel pins in the HEDL-P-60 subassembly in EBR-II. Initial selection of candidate advanced alloys was done using only nonfueled materials test results. However, to establish the performance characteristics of the candidate cladding alloys, i.e., dimensional stability and structural integrity under conditions of high neutron flux, elevated temperature, and applied stress, it was necessary to irradiate fuel pins under typical operating conditions. Fuel pins were clad with solution treated IN-706 and PE-16 and irradiated to peak fluences of 6.1 x 10 22 n/cm 2 (E > .1 MeV) and 8.8 x 10 22 n/cm 2 (E > .1 MeV) respectively. Fabrication and operating parameters for the fuel pins with the advanced cladding alloy candidates are summarized. Irradiation of HEDL-P-60 was interrupted with the breach of a pin with IN-706 cladding at 5.1 at % and the test was terminated with cladding breach in a pin with PE-16 cladding at 7.6 at %

Transient survivability of LMR oxide fuel pins

International Nuclear Information System (INIS)

Weber, E.T.; Pitner, A.L.; Bard, F.E.; Culley, G.E.; Hunter, C.W.

1986-01-01

Fuel pin integrity during transient events must be assessed for both the core design and safety analysis phases of a reactor project. A significant increase in the experience related to limits of integrity for oxide fuel pins in transient overpower events has been realized from testing of fuel pins irradiated in FFTF and PFR. Fourteen FFTF irradiated fuel pins were tested in TREAT, representing a range of burnups, overpower ramp rates and maximum overpower conditions. Results of these tests along with similar testing in the PFR/TREAT program, provide a demonstration of significant safety margins for oxide fuel pins. Useful information applied in analytical extrapolation of fuel pin test data have been developed from laboratory transient tests on irradiated fuel cladding (FCTT) and on unirradiated fuel pellet deformation. These refinements in oxide fuel transient performance are being applied in assessment of transient capabilities of long lifetime fuel designs using ferritic cladding

Probabilistic distributions of pin gaps within a wire-spaced fuel subassembly and sensitivities of the related uncertainties to pin gap

International Nuclear Information System (INIS)

Sakai, K.; Hishida, H.

1978-01-01

Probabilistic fuel pin gap distributions within a wire-spaced fuel subassembly and sensitivities of the related uncertainties to fuel pin gaps are discussed. The analyses consist mainly of expressing a local fuel pin gap in terms of sensitivity functions of the related uncertainties and calculating the corresponding probabilistic distribution through taking all the possible combinations of the distribution of uncertainties. The results of illustrative calculations show that with the reliability level of 0.9987, the maximum deviation of the pin gap at the cladding hot spot of a center fuel subassembly is 8.05% from its nominal value and the corresponding probabilistic pin gap distribution is shifted to the narrower side due to the external confinement of a pin bundle with a wrapper tube. (Auth.)

Cesium migration in LMFBR fuel pins

International Nuclear Information System (INIS)

Karnesky, R.A.; Jost, J.W.; Stone, I.Z.

1978-10-01

The factors affecting the axial migration of cesium in mixed oxide fuel pins and the effects of cesium migration on fuel pin performance are examined. The development and application of a correlated model which will predict the occurrence of cesium migration in a mixed oxide (75 w/o UO 2 + 25 w/o PuO 2 ) fuel pins over a wide range of fabrication and irradiation conditions are described

Calculation of fuel pin failure timing under LOCA conditions

International Nuclear Information System (INIS)

Jones, K.R.; Wade, N.L.; Siefken, L.J.; Straka, M.; Katsma, K.R.

1991-10-01

The objective of this research was to develop and demonstrate a methodology for calculation of the time interval between receipt of the containment isolation signals and the first fuel pin failure for loss-of-coolant accidents (LOCAs). Demonstration calculations were performed for a Babcock and Wilcox (B ampersand W) design (Oconee) and a Westinghouse (W) 4-loop design (Seabrook). Sensitivity studies were performed to assess the impacts of fuel pin burnup, axial peaking factor, break size, emergency core cooling system (ECCS) availability, and main coolant pump trip on these items. The analysis was performed using a four-code approach, comprised of FRAPCON-2, SCDAP/RELAP5/MOD3, TRAC-PF1/MOD1, and FRAP-T6. In addition to the calculation of timing results, this analysis provided a comparison of the capabilities of SCDAP/RELAP5/MOD3 with TRAC-PF1/MOD1 for large-break LOCA analysis. This paper discusses the methodology employed and the code development efforts required to implement the methodology. The shortest time intervals calculated between initiation of containment isolation and fuel pin failure were 11.4 s and 19.1 for the B ampersand W and W plants, respectively. The FRAP-T6 fuel pin failure times calculated using thermal-hydraulic data generated by SCDAP/RELAP5/MOD3 were more conservative than those calculated using data generated by TRAC-PF1/MOD1. 18 refs., 7 figs., 4 tabs

Automated fuel pin loading system

Science.gov (United States)

Christiansen, D.W.; Brown, W.F.; Steffen, J.M.

An automated loading system for nuclear reactor fuel elements utilizes a gravity feed conveyor which permits individual fuel pins to roll along a constrained path perpendicular to their respective lengths. The individual lengths of fuel cladding are directed onto movable transports, where they are aligned coaxially with the axes of associated handling equipment at appropriate production stations. Each fuel pin can be be reciprocated axially and/or rotated about its axis as required during handling steps. The fuel pins are inerted as a batch prior to welding of end caps by one of two disclosed welding systems.

Sodium Loop Safety Facility W-2 experiment fuel pin rupture detection system

International Nuclear Information System (INIS)

Hoffman, M.A.; Kirchner, T.L.; Meyers, S.C.

1980-05-01

The objective of the Sodium Loop Safety Facility (SLSF) W-2 experiment is to characterize the combined effects of a preconditioned full-length fuel column and slow transient overpower (TOP) conditions on breeder reactor (BR) fuel pin cladding failures. The W-2 experiment will meet this objective by providing data in two technological areas: (1) time and location of cladding failure, and (2) early post-failure test fuel behavior. The test involves a seven pin, prototypic full-length fast test reactor (FTR) fuel pin bundle which will be subjected to a simulated unprotected 5 cents/s reactivity transient overpower event. The outer six pins will provide the necessary prototypic thermal-hydraulic environment for the center pin

Robustness of pinning a general complex dynamical network

International Nuclear Information System (INIS)

Wang Lei; Sun Youxian

2010-01-01

This Letter studies the robustness problem of pinning a general complex dynamical network toward an assigned synchronous evolution. Several synchronization criteria are presented to guarantee the convergence of the pinning process locally and globally by construction of Lyapunov functions. In particular, if a pinning strategy has been designed for synchronization of a given complex dynamical network, then no matter what uncertainties occur among the pinned nodes, synchronization can still be guaranteed through the pinning. The analytical results show that pinning control has a certain robustness against perturbations on network architecture: adding, deleting and changing the weights of edges. Numerical simulations illustrated by scale-free complex networks verify the theoretical results above-acquired.

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.

Pinning Mechanisms in YBCO Tapes

CERN Document Server

Spera, Marcello; Ballarino, Amalia

2015-01-01

In this thesis work, a study on flux pinning mechanisms of commercial YBCO tapes is presented. This study has been performed via critical current characterization using transport (via direct I-V curves) and magnetization (via a Vibrating Sample Magnetometer) measurements. The latter ones turned out to be better concerning the comprehension of the pinning landscape of the provided samples, as a wider range of magnetic fields and temperatures is available for those measurements in the setup I used. The comparison of the experimental data with existing theoretical models allowed me to draw a picture of the pinning mechanisms underlying in each sample, and they turned out to be quite different one another. Moreover, for high-performance research tapes, another interesting feature has been found: the counterplay between the self-field critical current and the in-field one. Very well engineered artificial pinning structures limit the self-field critical current density due to the hi...

Open reading frame 176 in the photosynthesis gene cluster of Rhodobacter capsulatus encodes idi, a gene for isopentenyl diphosphate isomerase.

OpenAIRE

Hahn, F M; Baker, J A; Poulter, C D

1996-01-01

Isopentenyl diphosphate (IPP) isomerase catalyzes an essential activation step in the isoprenoid biosynthetic pathway. A database search based on probes from the highly conserved regions in three eukaryotic IPP isomerases revealed substantial similarity with ORF176 in the photosynthesis gene cluster in Rhodobacter capsulatus. The open reading frame was cloned into an Escherichia coli expression vector. The encoded 20-kDa protein, which was purified in two steps by ion exchange and hydrophobic...

Crystal structure of Pyrococcus furiosus phosphoglucose isomerase: Implications for substrate binding and catalysis

NARCIS (Netherlands)

Berrisford, J.M.; Akerboom, A.P.; Turnbull, A.P.; Geus, de D.; Sedelnikova, S.E.; Staton, I.; McLeod, C.W.; Verhees, C.H.; Oost, van der J.; Rice, D.W.; Baker, P.J.

2003-01-01

Phosphoglucose isomerase (PGI) catalyzes the reversible isomerization between D-fructose 6-phosphate and D-glucose 6-phosphate as part of the glycolytic pathway. PGI from the Archaea Pyrococcus furiosus (Pfu) was crystallized, and its structure was determined by x-ray diffraction to a 2-Angstrom

Fuel-pin cladding transient failure strain criterion

International Nuclear Information System (INIS)

Bard, F.E.; Duncan, D.R.; Hunter, C.W.

1983-01-01

A criterion for cladding failure based on accumulated strain was developed for mixed uranium-plutonium oxide fuel pins and used to interpret the calculated strain results from failed transient fuel pin experiments conducted in the Transient Reactor Test (TREAT) facility. The new STRAIN criterion replaced a stress-based criterion that depends on the DORN parameter and that incorrectly predicted fuel pin failure for transient tested fuel pins. This paper describes the STRAIN criterion and compares its prediction with those of the stress-based criterion

Pressure and Temperature of the Room 1 for the Pipe Break Accidents of the 3-Pin Fuel Test Loop

Energy Technology Data Exchange (ETDEWEB)

Park, S. K.; Chi, D. Y.; Sim, B. S.; Park, K. N.; Ahn, S. H.; Lee, J. M.; Lee, C. Y.; Kim, H. R

2005-08-15

This report deals with the prediction of the pressure and temperature of the room 1 for the pipe break accidents of the 3-pin fuel test loop. The 3-pin fuel test loop is an experimental facility for nuclear fuel tests at the operation conditions similar to those of PWR and CANDU power plants. Because the most processing systems of the 3-pin fuel test loop are placed in the room 1. The structural integrity of the room 1 should be evaluated for the postulated accident conditions. Therefore the pressures and temperatures of the room 1 needed for the structural integrity evaluation have been calculated by using MARS code. The pressures and temperatures of the room 1 have been calculated in various conditions such as the thermal hydraulic operation parameters, the locations of pipe break, and the thermal properties of the room 1 wall. It is assumed that the pipe break accident occurs in the letdown operation without regeneration, because the mass and energy release to the room 1 is expected to be the largest. As a result of the calculations the maximum pressure and temperature are predicted to be 208kPa and 369.2K(96.0 .deg. C) in case the heat transfer is considered in the room 1 wall. However the pressure and temperature are asymptotically 243kPa and 378.1K(104.9 .deg. C) assuming that the heat transfer does not occur in the room 1 wall.

Enhanced pinning in superconducting thin films with graded pinning landscapes

Science.gov (United States)

Motta, M.; Colauto, F.; Ortiz, W. A.; Fritzsche, J.; Cuppens, J.; Gillijns, W.; Moshchalkov, V. V.; Johansen, T. H.; Sanchez, A.; Silhanek, A. V.

2013-05-01

A graded distribution of antidots in superconducting a-Mo79Ge21 thin films has been investigated by magnetization and magneto-optical imaging measurements. The pinning landscape has maximum density at the sample border, decreasing linearly towards the center. Its overall performance is noticeably superior than that for a sample with uniformly distributed antidots: For high temperatures and low fields, the critical current is enhanced, whereas the region of thermomagnetic instabilities in the field-temperature diagram is significantly suppressed. These findings confirm the relevance of graded landscapes on the enhancement of pinning efficiency, as recently predicted by Misko and Nori [Phys. Rev. B 85, 184506 (2012)].

PINS Testing and Modification for Explosive Identification

International Nuclear Information System (INIS)

Seabury, E.H.; Caffrey, A.J.

2011-01-01

The INL's Portable Isotopic Neutron Spectroscopy System (PINS)1 non-intrusively identifies the chemical fill of munitions and sealed containers. PINS is used routinely by the U.S. Army, the Defense Threat Reduction Agency, and foreign military units to determine the contents of munitions and other containers suspected to contain explosives, smoke-generating chemicals, and chemical warfare agents such as mustard and nerve gas. The objects assayed with PINS range from softball-sized M139 chemical bomblets to 200 gallon DOT 500X ton containers. INL had previously examined2 the feasibility of using a similar system for the identification of explosives, and based on this proof-of-principle test, the development of a dedicated system for the identification of explosives in an improvised nuclear device appears entirely feasible. INL has been tasked by NNSA NA-42 Render Safe Research and Development with the development of such a system.

Evaluating Approaches to Rendering Braille Text on a High-Density Pin Display.

Science.gov (United States)

Morash, Valerie S; Russomanno, Alexander; Gillespie, R Brent; OModhrain, Sile

2017-10-13

Refreshable displays for tactile graphics are typically composed of pins that have smaller diameters and spacing than standard braille dots. We investigated configurations of high-density pins to form braille text on such displays using non-refreshable stimuli produced with a 3D printer. Normal dot braille (diameter 1.5 mm) was compared to high-density dot braille (diameter 0.75 mm) wherein each normal dot was rendered by high-density simulated pins alone or in a cluster of pins configured in a diamond, X, or square; and to "blobs" that could result from covering normal braille and high-density multi-pin configurations with a thin membrane. Twelve blind participants read MNREAD sentences displayed in these conditions. For high-density simulated pins, single pins were as quickly and easily read as normal braille, but diamond, X, and square multi-pin configurations were slower and/or harder to read than normal braille. We therefore conclude that as long as center-to-center dot spacing and dot placement is maintained, the dot diameter may be open to variability for rendering braille on a high density tactile display.

Performance of electrical contact pins near a nuclear explosion

International Nuclear Information System (INIS)

Ragan, C.E.; Silbert, M.G.; Ellis, A.N.; Robinson, E.E.; Daddario, M.J.

1977-09-01

The pressures attainable in equation-of-state studies using nuclear-explosion-driven shock waves greatly exceed those that can be reached in normal laboratory conditions. However, the diagnostic instrumentation must survive in the high-radiation environment present near such an explosion. Therefore, a set of experiments were fielded on the Redmud event to test the feasibility of using electrical contact pins in this environment. In these experiments a 60-cm-high shield of boron-lead was placed on the rack lid approximately 1 m from the device. A sample consisting of slabs of molybdenum and 238 U was placed on top of the shield, and twelve electrical contact pins were embedded to five different depths in the materials. Five different multiplexing-charging circuits were used for the pins, and a piezoelectric quartz gauge was placed on top of the uranium to obtain an estimate of the fission-energy deposition. All of the charged pins survived the radiation and produced signals indicating shock arrival. The uncertainty in determining the pin-closure time was approximately 3 ns. The signal from the quartz gauge corresponded to a pressure that was consistent with the calculated neutron fluence

FFTF fuel pin design bases and performance

International Nuclear Information System (INIS)

Cox, C.M.; Hanson, J.E.; Roake, W.E.; Slember, R.J.; Weber, C.E.; Millunzi, A.C.

1975-04-01

The FFTF fuel pin was conservatively designed to meet thermal and structural performance requirements in the categories normal operation, upset events, emergency events, and hypothetical, faulted events. The fuel pin operating limits consistent with these requirements were developed from a strong fuel pin irradiation testing program scoped to define the performance capability under relevant steady state and transient conditions. Comparison of the results of the irradiation testing program with design requirements indicates that the FFTF fuel pin can exceed its goal burnup of 80,000 MWd/MTM. (U.S.)

Ferromagnetic artificial pinning centers in multifilamentary superconducting wires

International Nuclear Information System (INIS)

Wang, J.Q.; Rizzo, N.D.; Prober, D.E.

1997-01-01

The authors fabricated multifilamentary NbTi wires with ferromagnetic (FM) artificial pinning centers (APCs) to enhance the critical current density (J c ) in magnetic fields. They used a bundle and draw technique to process the APC wires with either Ni or Fe as the pinning centers. Both wires produced higher J c in the high field range (5-9 T) than previous non-magnetic APC wires similarly processed, even though the authors have not yet optimized pin percentage. Using a magnetometer they found that the pins remained ferromagnetic for the wires with maximum J c . However, they did observe a substantial loss of FM material for the wires where the pin diameter approached 3 nm. Thus, they expect further enhancement of J c with better pin quality

Pinning control of complex networked systems synchronization, consensus and flocking of networked systems via pinning

CERN Document Server

Su, Housheng

2013-01-01

Synchronization, consensus and flocking are ubiquitous requirements in networked systems. Pinning Control of Complex Networked Systems investigates these requirements by using the pinning control strategy, which aims to control the whole dynamical network with huge numbers of nodes by imposing controllers for only a fraction of the nodes. As the direct control of every node in a dynamical network with huge numbers of nodes might be impossible or unnecessary, it’s then very important to use the pinning control strategy for the synchronization of complex dynamical networks. The research on pinning control strategy in consensus and flocking of multi-agent systems can not only help us to better understand the mechanisms of natural collective phenomena, but also benefit applications in mobile sensor/robot networks. This book offers a valuable resource for researchers and engineers working in the fields of control theory and control engineering.   Housheng Su is an Associate Professor at the Department of Contro...

Automation of FBTR fuel pin inspection using FPGA

International Nuclear Information System (INIS)

Khare, K.M.; Pai, Siddhesh; Pant, Brijesh; Sendhil Raja, S.; Gupta, P.K.

2011-01-01

A non-contact metrology system for inspection of FBTR fuel pins has been developed. The system consists of a stepper motors driven mechanism for orientation and positioning of FBTR fuel pin, a telecentric imaging system, absolute linear encoder with 0.1 μm resolution and a Field Programmable Gate Array (FPCA) based controller. The FBTR pin assembly is telecentrically illuminated from bottom by a red LED and its shadow graph is imaged using a CCD camera through telecentric imaging lens system. For system control and automation we have used a FPGA that has integrated soft picoblaze processor, X-θ axis motion controller, custom IPs for encoder data acquisition, synchronization circuit, RS485 interface along with other l/Os. Using the Graphical User Interface (GUI) on a PC the system is initialized at home position and the controller provides the trigger signal for start of data acquisition of CCD camera. CCD image of pin and the corresponding X-θ information is captured. After the acquisition of one set of images, the imaging module is moved with a step size pre-programmed to ensure proper stitching of acquired images. The GUI is programmed to analyze these X-θ Images to calculate the required parameters of the fuel pin like the diameter variation, pitch and bow. The details of the instrument and measurements made with it will be presented. (author)

Dynamic Phases in Driven Vortex Lattices in Superconductors with Periodic Pinning Arrays.

Science.gov (United States)

Reichhardt, C.; Olson, C. J.; Nori, F.

1997-03-01

In an extensive series of simulations of driven vortices interacting with periodic pinning arrays, an extremely rich variety of novel plastic flow phases, very distinct from those observed in random arrays, are found as a function of applied driving force. We show that signatures of the transitions between these different dynamical phases appear as pronounced jumps and dips in the I-V curves, coinciding with marked changes in the microscopic structure and flow behavior of the vortex lattice. When the number of vortices is greater than the number of pinning sites, we observe up to six distinct dynamical phases, including a pinned phase, a flow of interstitial vortices between pinned vortices, a disordered flow, a 1D flow along the pinning rows, and a homogeneous flow. By varying a wide range of microscopic pinning parameters, including pinning strength, size, density, and degree of ordering, as well as varying temperature and commensurability, we obtain a series of dynamic phase diagrams. nori>A short video will also be presented to highlight these different dynamic phases.

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

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

Directory of Open Access Journals (Sweden)

Hirokazu Tanaka

2013-05-01

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

Nano-engineered pinning centres in YBCO superconducting films

Energy Technology Data Exchange (ETDEWEB)

Crisan, A., E-mail: adrian.crisan@infim.ro [National Institute for Materials Physics Bucharest, 105 bis Atomistilor Str., 077125 Magurele (Romania); School of Metallurgy and Materials, University of Birmingham, Edgbaston, B15 2TT Birmingham (United Kingdom); Dang, V.S. [School of Metallurgy and Materials, University of Birmingham, Edgbaston, B15 2TT Birmingham (United Kingdom); Nano and Energy Center, VNU Hanoi University of Science, 334 Nguyen Trai, Thanh Xuan, Hanoi (Viet Nam); Mikheenko, P. [School of Metallurgy and Materials, University of Birmingham, Edgbaston, B15 2TT Birmingham (United Kingdom); Department of Physics, University of Oslo, P.O. Box 1048 Blindern, N-0316 Oslo (Norway)

2017-02-15

Highlights: • Power applications of YBCO films/coated conductors in technological relevant magnetic fields requires nano-engineered pinning centre. • Three approaches have been proposed: substrate decoration, quasi-multilayers, and targets with secondary phase nano-inclusions. • Combination of all three approaches greatly increased critical current in YBCO films. • Bulk pinning force, pinning potential, and critical current density are estimated and discussed in relation with the type and strength of pinning centres related to the defects evidenced by Transmission Electron Microscopy. - Abstract: For practical applications of superconducting materials in applied magnetic fields, artificial pinning centres in addition to natural ones are required to oppose the Lorentz force. These pinning centres are actually various types of defects in the superconductor matrix. The pinning centres can be categorised on their dimension (volume, surface or point) and on their character (normal cores or Δκ cores). Different samples have been produced by Pulsed Laser Deposition, with various thicknesses, temperatures and nanostructured additions to the superconducting matrix. They have been characterized by SQUID Magnetic Properties Measurement System and Physical Properties Measurement System, as well as by Transmission Electron Microscopy (TEM). Correlations between pinning architecture, TEM images, and critical currents at various fields and field orientations will be shown for a large number of YBa{sub 2}Cu{sub 3}O{sub x} films with various types and architectures of artificial pinning centres.

Role of protein disulfide isomerase and other thiol-reactive proteins in HIV-1 envelope protein-mediated fusion

International Nuclear Information System (INIS)

Ou Wu; Silver, Jonathan

2006-01-01

Cell-surface protein disulfide isomerase (PDI) has been proposed to promote disulfide bond rearrangements in HIV-1 envelope protein (Env) that accompany Env-mediated fusion. We evaluated the role of PDI in ways that have not been previously tested by downregulating PDI with siRNA and by overexpressing wild-type or variant forms of PDI in transiently and stably transfected cells. These manipulations, as well as treatment with anti-PDI antibodies, had only small effects on infection or cell fusion mediated by NL4-3 or AD8 strains of HIV-1. However, the cell-surface thiol-reactive reagent 5, 5'-dithiobis(2-nitrobenzoic acid) (DTNB) had a much stronger inhibitory effect in our system, suggesting that cell-surface thiol-containing molecules other than PDI, acting alone or in concert, have a greater effect than PDI on HIV-1 Env-mediated fusion. We evaluated one such candidate, thioredoxin, a PDI family member reported to reduce a labile disulfide bond in CD4. We found that the ability of thioredoxin to reduce the disulfide bond in CD4 is enhanced in the presence of HIV-1 Env gp120 and that thioredoxin also reduces disulfide bonds in gp120 directly in the absence of CD4. We discuss the implications of these observations for identification of molecules involved in disulfide rearrangements in Env during fusion

Tailoring the vortex pinning strength of YBCO thin films by systematic incorporation of hybrid artificial pinning centers

International Nuclear Information System (INIS)

Jha, Alok K; Matsumoto, Kaname; Horide, Tomoya; Saini, Shrikant; Mele, Paolo; Ichinose, Ataru; Yoshida, Yutaka; Awaji, Satoshi

2015-01-01

The effect of hybrid (columnar and spherical together) artificial pinning centers (APCs) on the vortex pinning properties of YBa 2 Cu 3 O 7−δ (YBCO) thin films is investigated in detail on the basis of variation of critical current density (J C ) with applied magnetic field and also with the orientation of the applied magnetic field at 65 K and 77 K. Premixed YBCO + BaSnO 3 composite targets are used for the deposition of the YBCO films which consist of self-assembled BaSnO 3 nanocolumns (1D APCs); on the other hand, for the deposition of the YBCO films with hybrid APCs (BaSnO 3 nanocolumns together with Y 2 O 3 nanoparticles), the surface of the premixed YBCO + BaSnO 3 composite targets are modified by putting a thin Y 2 O 3 sectored piece on the premixed YBCO + BaSnO 3 composite targets by means of silver paste. F pmax value increases systematically with incorporation of 1D and 1D and 3D APCs and it also shifts towards higher applied magnetic fields. Films with 1D APCs exhibit a strong J C peak at Θ = 0° (H//c-axis) whereas films consisting of hybrid APCs exhibit enhanced J C at all the investigated angular regimes. A possible mechanism of vortex pinning in samples with hybrid APCs is also discussed suggesting the role of 1D and 3D APCs. (paper)

Investigation on Bond-Slip Behavior of Z-Pin Interfaces in X-Cor® Sandwich Structures Using Z-Pin Pull-Out Test

Science.gov (United States)

Shan, Hangying; Xiao, Jun; Chu, Qiyi

2018-05-01

The Z-Pin interfacial bond properties play an important role in the structural performance of X-Cor® sandwich structures. This paper presents an experimental investigation on bond-slip behavior of Z-Pin interfaces using Z-Pin pull-out test. Based on the experimental data the whole Z-Pin pull-out process consists of three stages: initial bonding, debonding and frictional sliding. Comparative experimental study on the influence of design parameters on bond-slip behavior of Z-Pin interfaces has also been performed. Numerical analyses were conducted with the ABAQUS finite element (FE) program to simulate the Z-Pins bond-slip response of the pull-out test. The Z-Pins interfacial bond-slip behavior was implemented using nonlinear spring elements characterized with the constitutive relation from experimental results. Numerical results were validated by comparison with experimental data, and reasonably good agreement was achieved between experimental and analytical pull-out force-slip curves.

Repeat biopsy in patients with initial diagnosis of PIN; La biopsia ripetuta nei pazienti con diagnosi iniziale di PIN

Energy Technology Data Exchange (ETDEWEB)

De Matteis, Massimo [Azienda Ospedaliera Policlinico S. Orsola-Malpighi, Bologna (Italy). UO Radiologia Albertoni; Poggi, Cristina; De Martino, Antonietta; Pavlica, Pietro [Azienda Ospedaliera Policlinico S. Orsola-Malpighi, Bologna (Italy). UO Radiologia Palagi, Dipartimento area radiologica; Corti, Barbara [Azienda Ospedaliera Policlinico S. Orsola-Malpighi, Bologna (Italy). UO Anatomia ed istologia patologica, Dipartimento oncologico ed ematologico; Barozzi, Libero [Azienda Ospedaliera Policlinico S. Orsola-Malpighi, Bologna (Italy). UO Radiologia d' urgenza, Dipartimento emergenze ed accettazione

2005-09-15

Purpose. Prostatic intra-epithelial neoplasia (PIN) is considered a pre-malignant lesion and the main precursor of invasive prostatic adenocarcinoma. A PIN diagnosis established by prostate needle biopsy poses a difficult clinical management. problem. We retrospectively reviewed our three-year experience in order to identify criteria for referring patients to repeat biopsy. Materials and methods. We reviewed the repeat biopsy records of 72 patients in whom PIN had been detected on initial US-guided needle biopsy of the prostate. All the patients had a minimum of 6 biopsy cores taken, and they all had PSA > 4 ng/ml. Results. Adenocarcinoma was detected in 15 patients out of 50 (30%) with an initial diagnosis of low-grade PIN and in 10 patients out of 22 (45.4%) with high grade PIN, in 7 out of 18 (39%) in whom PSA levels had decreased during the observation interval, in 16 patients out of 46 (35%) in whom the PSA had increased and in 2 patients out of 8 (25%) with stable PSA. Conclusions. Our results seem to confirm that PIN can be considered a precursor of prostatic adenocarcinoma or a histological alteration often associated with it. Patients with low-grade PIN and particularly those with high-grade PIN should be regularly subjected to repeat biopsy at short intervals due to the high frequency of the final diagnosis of carcinoma. No agreement has been reached on the time interval between the first and the second biopsy. The PSA changes during the observation period are not a statistically significant parameter to suggest the repetition of prostatic biopsy. [Italian] Scopo. La neoplasia prostatica intraepiteliale (PIN) e considerata una lesione premaligna ed il precursore principale dell'adenocarcinoma prostatico infiltrante. La diagnosi di PIN ottenuta con l'agobiopsia della prostata rappresenta un difficile problema gestionale clinico. In una valutazione retrospettiva della nostra esperienza di 3 anni si e cercato di individuare i criteri che possano

Vortex pinning and creep experiments

International Nuclear Information System (INIS)

Kes, P.H.

1991-01-01

A brief review of basic flux-pinning and flux-creep ingredients and a selection of experimental results on high-temperature-superconductivity compounds is presented. Emphasis is put on recent results and on those properties which are central to the emerging understanding of the flux-pinning and flux-creep mechanisms of these fascinating materials

Parasitic nematodes modulate PIN-mediated auxin transport to facilitate infection.

Directory of Open Access Journals (Sweden)

Wim Grunewald

2009-01-01

Full Text Available Plant-parasitic nematodes are destructive plant pathogens that cause significant yield losses. They induce highly specialized feeding sites (NFS in infected plant roots from which they withdraw nutrients. In order to establish these NFS, it is thought that the nematodes manipulate the molecular and physiological pathways of their hosts. Evidence is accumulating that the plant signalling molecule auxin is involved in the initiation and development of the feeding sites of sedentary plant-parasitic nematodes. Intercellular transport of auxin is essential for various aspects of plant growth and development. Here, we analysed the spatial and temporal expression of PIN auxin transporters during the early events of NFS establishment using promoter-GUS/GFP fusion lines. Additionally, single and double pin mutants were used in infection studies to analyse the role of the different PIN proteins during cyst nematode infection. Based on our results, we postulate a model in which PIN1-mediated auxin transport is needed to deliver auxin to the initial syncytial cell, whereas PIN3 and PIN4 distribute the accumulated auxin laterally and are involved in the radial expansion of the NFS. Our data demonstrate that cyst nematodes are able to hijack the auxin distribution network in order to facilitate the infection process.

Parasitic nematodes modulate PIN-mediated auxin transport to facilitate infection.

Science.gov (United States)

Grunewald, Wim; Cannoot, Bernard; Friml, Jirí; Gheysen, Godelieve

2009-01-01

Plant-parasitic nematodes are destructive plant pathogens that cause significant yield losses. They induce highly specialized feeding sites (NFS) in infected plant roots from which they withdraw nutrients. In order to establish these NFS, it is thought that the nematodes manipulate the molecular and physiological pathways of their hosts. Evidence is accumulating that the plant signalling molecule auxin is involved in the initiation and development of the feeding sites of sedentary plant-parasitic nematodes. Intercellular transport of auxin is essential for various aspects of plant growth and development. Here, we analysed the spatial and temporal expression of PIN auxin transporters during the early events of NFS establishment using promoter-GUS/GFP fusion lines. Additionally, single and double pin mutants were used in infection studies to analyse the role of the different PIN proteins during cyst nematode infection. Based on our results, we postulate a model in which PIN1-mediated auxin transport is needed to deliver auxin to the initial syncytial cell, whereas PIN3 and PIN4 distribute the accumulated auxin laterally and are involved in the radial expansion of the NFS. Our data demonstrate that cyst nematodes are able to hijack the auxin distribution network in order to facilitate the infection process.

L-Rhamnose isomerase and its use for biotechnological production of rare sugars.

Science.gov (United States)

Xu, Wei; Zhang, Wenli; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

2016-04-01

L-Rhamnose isomerase (L-RI, EC 5.3.1.14), catalyzing the isomerization between L-rhamnose and L-rhamnulose, plays an important role in microbial L-rhamnose metabolism and thus occurs in a wide range of microorganisms. It attracts more and more attention because of its broad substrate specificity and its great potential in enzymatic production of various rare sugars. In this article, the enzymatic properties of various reported L-RIs were compared in detail, and their applications in the production of L-rhamnulose and various rare sugars including D-allose, D-gulose, L-lyxose, L-mannose, L-talose, and L-galactose were also reviewed.

Nuclear fuel pin controlled failure device

International Nuclear Information System (INIS)

Schlenker, L.D.

1975-01-01

Each fuel pin of a fuel assembly for a water-cooled nuclear reactor is provided with means for rupturing the cladding tube at a predetermined location if an abnormal increase in pressure of the gases present occurs due to a loss-of-coolant accident. Preferably all such rupture means are oriented to minimize the hydraulic resistance to the flow of emergency core coolant such as all rupture means pointing in the same direction. Rupture means may be disposed at different elevations in adjacent fuel pins and, further, fuel pins may be provided with two or more rupture means, one of which is in the upper portion of the fuel pin. Rupture means are mechanical as by providing a locally weakened condition of a controlled nature in the cladding. (U.S.)

Silicide Schottky Contacts to Silicon: Screened Pinning at Defect Levels

Energy Technology Data Exchange (ETDEWEB)

Drummond, T.J.

1999-03-11

Silicide Schottky contacts can be as large as 0.955 eV (E{sub v} + 0.165 eV) on n-type silicon and as large as 1.05 eV (E{sub c} {minus} 0.07 eV) on p-type silicon. Current models of Schottky barrier formation do not provide a satisfactory explanation of occurrence of this wide variation. A model for understanding Schottky contacts via screened pinning at defect levels is presented. In the present paper it is shown that most transition metal silicides are pinned approximately 0.48 eV above the valence band by interstitial Si clusters. Rare earth disilicides pin close to the divacancy acceptor level 0.41 eV below the conduction band edge while high work function silicides of Ir and Pt pin close to the divacancy donor level 0.21 eV above the valence band edge. Selection of a particular defect pinning level depends strongly on the relative positions of the silicide work function and the defect energy level on an absolute energy scale.

Parallel two-phase-flow-induced vibrations in fuel pin model

International Nuclear Information System (INIS)

Hara, Fumio; Yamashita, Tadashi

1978-01-01

This paper reports the experimental results of vibrations of a fuel pin model -herein meaning the essential form of a fuel pin from the standpoint of vibration- in a parallel air-and-water two-phase flow. The essential part of the experimental apparatus consisted of a flat elastic strip made of stainless steel, both ends of which were firmly supported in a circular channel conveying the two-phase fluid. Vibrational strain of the fuel pin model, pressure fluctuation of the two-phase flow and two-phase-flow void signals were measured. Statistical measures such as power spectral density, variance and correlation function were calculated. The authors obtained (1) the relation between variance of vibrational strain and two-phase-flow velocity, (2) the relation between variance of vibrational strain and two-phase-flow pressure fluctuation, (3) frequency characteristics of variance of vibrational strain against the dominant frequency of the two-phase-flow pressure fluctuation, and (4) frequency characteristics of variance of vibrational strain against the dominant frequency of two-phase-flow void signals. The authors conclude that there exist two kinds of excitation mechanisms in vibrations of a fuel pin model inserted in a parallel air-and-water two-phase flow; namely, (1) parametric excitation, which occurs when the fundamental natural frequency of the fuel pin model is related to the dominant travelling frequency of water slugs in the two-phase flow by the ratio 1/2, 1/1, 3/2 and so on; and (2) vibrational resonance, which occurs when the fundamental frequency coincides with the dominant frequency of the two-phase-flow pressure fluctuation. (auth.)

Enzymatic conversion of D-galactose to D-tagatose: heterologous expression and characterisation of a thermostable L-arabinose isomerase from Thermoanaerobacter mathranii.

Science.gov (United States)

Jørgensen, F; Hansen, O C; Stougaard, P

2004-06-01

The ability to convert D-galactose into D-tagatose was compared among a number of bacterial L-arabinose isomerases ( araA). One of the most efficient enzymes, from the anaerobic thermophilic bacterium Thermoanaerobacter mathranii, was produced heterologously in Escherichia coli and characterised. Amino acid sequence comparisons indicated that this enzyme is only distantly related to the group of previously known araA sequences in which the sequence similarity is evident. The substrate specificity and the Michaelis-Menten constants of the enzyme determined with L-arabinose, D-galactose and D-fucose also indicated that this enzyme is an unusual, versatile L-arabinose isomerase which is able to isomerise structurally related sugars. The enzyme was immobilised and used for production of D-tagatose at 65 degrees C. Starting from a 30% solution of D-galactose, the yield of D-tagatose was 42% and no sugars other than D-tagatose and D-galactose were detected. Direct conversion of lactose to D-tagatose in a single reactor was demonstrated using a thermostable beta-galactosidase together with the thermostable L-arabinose isomerase. The two enzymes were also successfully combined with a commercially available glucose isomerase for conversion of lactose into a sweetening mixture comprising lactose, glucose, galactose, fructose and tagatose.

Biochemical function of typical and variant Arabidopsis thaliana U-box E3 ubiquitin-protein ligases.

Science.gov (United States)

Wiborg, Jakob; O'Shea, Charlotte; Skriver, Karen

2008-08-01

The variance of the U-box domain in 64 Arabidopsis thaliana (thale cress) E3s (ubiquitin-protein ligases) was used to examine the interactions between E3s and E2s (ubiquitin-conjugating enzymes). E2s and E3s are components of the ubiquitin protein degradation pathway. Seven U-box proteins were analysed for their ability to ubiquitinate proteins in vitro in co-operation with different E2s. All U-box domains exhibited ubiquitination activity and interacted productively with UBC4/5-type E2s. Three and four of the U-box domains mediated ubiquitin addition in the presence of UBC13 and UBC7 E2s respectively, but no productive interaction was observed with the UBC15 E2 tested. The activity of AtPUB54 [Arabidopsis thaliana (thale cress) plant U-box 54 protein] was dependent on Trp(266) in the E2-binding cleft, and the E2 selectivity was changed by substitution of this position. The function of the distant U-box protein, AtPUB49, representing a large family of eukaryotic proteins containing a U-box linked to a cyclophilin-like peptidyl-prolyl cis-trans isomerase domain, was characterized biochemically. AtPUB49 functioned both as a prolyl isomerase and a chaperone by catalysing cis-trans isomerization of peptidyl-prolyl bonds and dissolving protein aggregates. In conclusion, both typical and atypical Arabidopsis U-box proteins were active E3s. The overlap in the E3/E2 selectivity suggests that in vivo specificity is not determined only by the E3-E2 interactions, but also by other parameters, e.g. co-existence or interactions with additional domains. The biochemical functions of AtPUB49 suggest that the protein can be involved in folding or degradation of protein substrates. Similar functions can also be retained within a protein complex with separate chaperone and U-box proteins.

Use of a modified transfixation pin cast for treatment of comminuted phalangeal fractures in horses.

Science.gov (United States)

Rossignol, Fabrice; Vitte, Amélie; Boening, Josef

2014-01-01

To (1) report a modified transfixation pin cast technique, using dorsal recumbency for fracture reduction, distal positioning of the pins in the epiphysis and distal metaphysis, and a hybrid cast, combining plaster of Paris (POP) and fiberglass casting, and (2) report outcome in 11 adult horses. Case series. Adult horses (n = 11) with comminuted phalangeal fractures. Horses were anesthetized and positioned in dorsal recumbency. The phalangeal fracture was reduced by limb traction using a cable attached to the hoof. Screw fixation in lag fashion of fracture fragments was performed when possible. Transfixation casting was performed using two 6.3 mm positive profile centrally threaded pins with the 1st pin placed in the epiphysis of the metacarpus/tarsus at the center of, or slightly proximal to, the condylar fossa and the 2nd one 3-4 cm proximal. A hybrid cast was applied. Forelimbs were involved in 9 horses and the hind limb in 2. Pins were maintained for a minimum of 6 weeks. No pin loosening was observed at the time of removal (6-8 weeks). A pony fractured the distal aspect of the metacarpus at the proximal pin. Nine horses survived (82%); none of the horses developed septic arthritis despite the distal location of the distal pin, close to the fetlock joint. This modified transfixation pin casting technique was associated with good pin longevity and could reduce the risk of secondary pin hole fractures and pin loosening. © Copyright 2013 by The American College of Veterinary Surgeons.

Propagation of spiral waves pinned to circular and rectangular obstacles.

Science.gov (United States)

Sutthiopad, Malee; Luengviriya, Jiraporn; Porjai, Porramain; Phantu, Metinee; Kanchanawarin, Jarin; Müller, Stefan C; Luengviriya, Chaiya

2015-05-01

We present an investigation of spiral waves pinned to circular and rectangular obstacles with different circumferences in both thin layers of the Belousov-Zhabotinsky reaction and numerical simulations with the Oregonator model. For circular objects, the area always increases with the circumference. In contrast, we varied the circumference of rectangles with equal areas by adjusting their width w and height h. For both obstacle forms, the propagating parameters (i.e., wavelength, wave period, and velocity of pinned spiral waves) increase with the circumference, regardless of the obstacle area. Despite these common features of the parameters, the forms of pinned spiral waves depend on the obstacle shapes. The structures of spiral waves pinned to circles as well as rectangles with the ratio w/h∼1 are similar to Archimedean spirals. When w/h increases, deformations of the spiral shapes are observed. For extremely thin rectangles with w/h≫1, these shapes can be constructed by employing semicircles with different radii which relate to the obstacle width and the core diameter of free spirals.

JMCT Monte Carlo simulation analysis of full core PWR Pin-By-Pin and shielding

International Nuclear Information System (INIS)

Deng, L.; Li, G.; Zhang, B.; Shangguan, D.; Ma, Y.; Hu, Z.; Fu, Y.; Li, R.; Hu, X.; Cheng, T.; Shi, D.

2015-01-01

This paper describes the application of the JMCT Monte Carlo code to the simulation of Kord Smith Challenge H-M model, BEAVRS model and Chinese SG-III model. For H-M model, the 6.3624 millions tally regions and the 98.3 billion neutron histories do. The detailed pin flux and energy deposition densities obtain. 95% regions have less 1% standard deviation. For BEAVRS model, firstly, we performed the neutron transport calculation of 398 axial planes in the Hot Zero Power (HZP) status. Almost the same results with MC21 and OpenMC results are achieved. The detailed pin-power density distribution and standard deviation are shown. Then, we performed the calculation of ten depletion steps in 30 axial plane cases. The depletion regions exceed 1.5 million and 12,000 processors uses. Finally, the Chinese SG-III laser model is simulated. The neutron and photon flux distributions are given, respectively. The results show that the JMCT code well suits for extremely large reactor and shielding simulation. (author)

Heat transfer in a fuel pin shipping container

International Nuclear Information System (INIS)

Ingham, J.G.

1980-01-01

Maximum cladding temperatures occur when the IDENT 1578 fuel pin shipping container is installed in the T-3 Cask. The maximum allowable cladding temperature of 800 0 F is reached when the rate of energy deposited in the 19-pin basket reaches 400 watts. Since 45% of the energy which is generated in the fuel escapes the 19-pin basket without being deposited, mostly gamma energy, the maximum allowable rate of heat generation is 400/.55 = 727 watts. Similarly, the maximum allowable cladding temperature of 800 0 F is reached when the rate of energy deposited in the 40-pin basket reaches 465 watts. Since 33% of the energy which is generated in the fuel escapes the 40-pin basket without being deposited, mostly gamma energy, the maximum allowable rate of heat generation is 465/.66 = 704 watts. The IDENT 1578 fuel pin shipping container therefore meets its thermal design criteria. IDENT 1578 can handle fuel pins with a decay heat load of 600 watts while maintaining the maximum fuel pin cladding temperature below 800 0 F. The emissivities which were determined from the test results for the basket tubes and container are relatively low and correspond to new, shiny conditions. As the IDENT 1578 container is exposed to high temperatures for extended periods of time during the transportation of fuel pins, the emissivities will probably increase. This will result in reduced temperatures

Electro-optical fuel pin identification system

International Nuclear Information System (INIS)

Kirchner, T.L.

1978-09-01

A prototype Electro-Optical Fuel Pin Identification System referred to as the Fuel Pin Identification System (FPIS) has been developed by the Hanford Engineering Development Laboratory (HEDL) in support of the Fast Flux Test Facility (FFTF) presently under construction at HEDL. The system is designed to remotely read an alpha-numeric identification number that is roll stamped on the top of the fuel pin end cap. The prototype FPIS consists of four major subassemblies: optical read head, digital compression electronics, video display, and line printer

Novel Roles of the Non-catalytic Elements of Yeast Protein-disulfide Isomerase in Its Interplay with Endoplasmic Reticulum Oxidoreductin 1*

Science.gov (United States)

Niu, Yingbo; Zhang, Lihui; Yu, Jiaojiao; Wang, Chih-chen; Wang, Lei

2016-01-01

The formation of disulfide bonds in the endoplasmic reticulum (ER) of eukaryotic cells is catalyzed by the sulfhydryl oxidase, ER oxidoreductin 1 (Ero1), and protein-disulfide isomerase (PDI). PDI is oxidized by Ero1 to continuously introduce disulfides into substrates, and feedback regulates Ero1 activity by manipulating the regulatory disulfides of Ero1. In this study we find that yeast Ero1p is enzymatically active even with its regulatory disulfides intact, and further activation of Ero1p by reduction of the regulatory disulfides requires the reduction of non-catalytic Cys90-Cys97 disulfide in Pdi1p. The principal client-binding site in the Pdi1p b′ domain is necessary not only for the functional Ero1p-Pdi1p disulfide relay but also for the activation of Ero1p. We also demonstrate by complementary activation assays that the regulatory disulfides in Ero1p are much more stable than those in human Ero1α. These new findings on yeast Ero1p-Pdi1p interplay reveal significant differences from our previously identified mode of human Ero1α-PDI interplay and provide insights into the evolution of the eukaryotic oxidative protein folding pathway. PMID:26846856

Development of disassembly and pin chopping technology for FBR spent fuels

International Nuclear Information System (INIS)

Kobayashi, Tsuguyuki; Namba, Takashi; Kawabe, Yukinari; Washiya, Tadahiro

2008-01-01

Japan Atomic Power Company (JAPC) and Japan Atomic Energy Agency (JAEA) have been developing fuel disassembly and fuel pin chopping systems for a future Japanese commercial FBR. At first, the wrapper tube is cut by the slit-cut to pull it out, then the fuel pins are cut by the crop-cut at their end-plugs to separate them from the entrance nozzle. The pins are transferred to the magazine of the chopping machine. A series of tests were performed to develop this procedure. As the result of mechanical cutting tests, the CBN wheel was selected. The slit-cut tests were carried out to evaluated the cutting performance of the wheel. The wrapper tube is normally slit-cut in the circumferential direction. One CBN wheel could cut more than 5 fuel assemblies in this direction. The slit-cut in the axial direction is prepared as provision when the tube is difficult to put out. More work is needed to cut 5mm thick PNC-FMS plate in this direction without damaging the pins beneath it. As the result of the crop-cut tests of end-plugs made of ODS steel, the CBN wheel could cut the 61 pin bundle by two strokes. More work is needed to cut the 217 pin bundle. Fuel pin handling tests were performed to transfer them from the disassembly machine to the chopping machine. The Saucer tray was selected to receive the disassembled pins. All the pins were transferred and loaded into a magazine of the chopping machine. Fuel pin loading tests were conducted to optimize the magazine configuration to make the chopping length within 1.0±0.5 cm. In order to decrease the disturbance during chopping, the width of the magazine was adjusted to be 12 cm and installation of a height adjuster is favourable to control the free space above the pins. (author)

Serviceability of rod ceramic fuel pins on motoring conditions of FTP or NEMF reactor

International Nuclear Information System (INIS)

Deryavko, I.I.

2004-01-01

The operation conditions of rod ceramic fuel pins in the running hydrogen-cooled technological canals of FTP or NEMF reactor on the motoring conditions are considered. The available postreactor researches of the fuel pins are presented and the additional postreactor researches of fuel pins, tested on this mode in IVG.1 and IRGIT reactors, are carried out. The fuel pins serviceability on motoring conditions of FTP or NEF reactor operation is concluded. (author)

Sodium Loop Safety Facility W-2 experiment fuel pin rupture detection system. [LMFBR

Energy Technology Data Exchange (ETDEWEB)

Hoffman, M.A.; Kirchner, T.L.; Meyers, S.C.

1980-05-01

The objective of the Sodium Loop Safety Facility (SLSF) W-2 experiment is to characterize the combined effects of a preconditioned full-length fuel column and slow transient overpower (TOP) conditions on breeder reactor (BR) fuel pin cladding failures. The W-2 experiment will meet this objective by providing data in two technological areas: (1) time and location of cladding failure, and (2) early post-failure test fuel behavior. The test involves a seven pin, prototypic full-length fast test reactor (FTR) fuel pin bundle which will be subjected to a simulated unprotected 5 cents/s reactivity transient overpower event. The outer six pins will provide the necessary prototypic thermal-hydraulic environment for the center pin.

Radon measurements with a PIN photodiode

International Nuclear Information System (INIS)

Martin-Martin, A.; Gutierrez-Villanueva, J.L.; Munoz, J.M.; Garcia-Talavera, M.; Adamiec, G.; Iniguez, M.P.

2006-01-01

Silicon photodiodes are well suited to detect alphas coming from different sources as neutron reactions or radon daughters. In this work a radon in air detecting device, using an 18x18 mm silicon PIN photodiode is studied. The ionized airborne decay products formed during radon diffusion were focused by an accelerating high voltage to the PIN surface. Several conducting rings were disposed inside a cylindrical PVC vessel in such a way that they reproduced the electric field created by a punctual charge located behind PIN position. Alpha spectra coming from the neutral and ionized species deposited on the PIN surface, dominated by 218 Po and 214 Po progeny peaks, were recorded for varying conditions. Those include radon concentration from a Pylon source, high voltage (thousands of volts) and PIN inverse bias voltage. Different parameters such as temperature and humidity were also registered during data acquisition. The increase in the particle collection efficiency with respect to zero electric field was compared with the corresponding to a parallel plates configuration. A discussion is made in terms of the most appropriate voltages for different radon concentrations

Interpretation of NMR relaxation properties of Pin1, a two-domain protein, based on Brownian dynamic simulations

International Nuclear Information System (INIS)

Bernado, Pau; Fernandes, Miguel X.; Jacobs, Doris M.; Fiebig, Klaus; Garcia de la Torre, Jose; Pons, Miquel

2004-01-01

Many important proteins contain multiple domains connected by flexible linkers. Inter-domain motion is suggested to play a key role in many processes involving molecular recognition. Heteronuclear NMR relaxation is sensitive to motions in the relevant time scales and could provide valuable information on the dynamics of multi-domain proteins. However, the standard analysis based on the separation of global tumbling and fast local motions is no longer valid for multi-domain proteins undergoing internal motions involving complete domains and that take place on the same time scale than the overall motion.The complexity of the motions experienced even for the simplest two-domain proteins are difficult to capture with simple extensions of the classical Lipari-Szabo approach. Hydrodynamic effects are expected to dominate the motion of the individual globular domains, as well as that of the complete protein. Using Pin1 as a test case, we have simulated its motion at the microsecond time scale, at a reasonable computational expense, using Brownian Dynamic simulations on simplified models. The resulting trajectories provide insight on the interplay between global and inter-domain motion and can be analyzed using the recently published method of isotropic Reorientational Mode Dynamics which offer a way of calculating their contribution to heteronuclear relaxation rates. The analysis of trajectories computed with Pin1 models of different flexibility provides a general framework to understand the dynamics of multi-domain proteins and explains some of the observed features in the relaxation rate profile of free Pin1

Interpretation of NMR relaxation properties of Pin1, a two-domain protein, based on Brownian dynamic simulations

Energy Technology Data Exchange (ETDEWEB)

Bernado, Pau [Institut de Biologie Structurale, Jean Pierre Ebel (France); Fernandes, Miguel X. [Universidad de Murcia, Departamento de Quimica Fisica, Facultad de Quimica (Spain); Jacobs, Doris M. [Johann Wolfgang Goethe-Universitaet Frankfurt, Institut fuer Organische Chemie und Chemische Biologie (Germany); Fiebig, Klaus [Affinium Pharmaceuticals (Canada); Garcia de la Torre, Jose [Universidad de Murcia, Departamento de Quimica Fisica, Facultad de Quimica (Spain); Pons, Miquel [Laboratori de RMN de Biomolecules, Parc Cientific de Barcelona (Spain)], E-mail: mpons@ub.edu

2004-05-15

Many important proteins contain multiple domains connected by flexible linkers. Inter-domain motion is suggested to play a key role in many processes involving molecular recognition. Heteronuclear NMR relaxation is sensitive to motions in the relevant time scales and could provide valuable information on the dynamics of multi-domain proteins. However, the standard analysis based on the separation of global tumbling and fast local motions is no longer valid for multi-domain proteins undergoing internal motions involving complete domains and that take place on the same time scale than the overall motion.The complexity of the motions experienced even for the simplest two-domain proteins are difficult to capture with simple extensions of the classical Lipari-Szabo approach. Hydrodynamic effects are expected to dominate the motion of the individual globular domains, as well as that of the complete protein. Using Pin1 as a test case, we have simulated its motion at the microsecond time scale, at a reasonable computational expense, using Brownian Dynamic simulations on simplified models. The resulting trajectories provide insight on the interplay between global and inter-domain motion and can be analyzed using the recently published method of isotropic Reorientational Mode Dynamics which offer a way of calculating their contribution to heteronuclear relaxation rates. The analysis of trajectories computed with Pin1 models of different flexibility provides a general framework to understand the dynamics of multi-domain proteins and explains some of the observed features in the relaxation rate profile of free Pin1.

Crystal Structure of Escherichia coli L-Arabinose Isomerase (ECAI), The Putative Target of Biological Tagatose Production

Energy Technology Data Exchange (ETDEWEB)

Manjasetty,B.; Chance, M.

2006-01-01

Escherichia coli L-arabinose isomerase (ECAI; EC 5.3.1.4) catalyzes the isomerization of L-arabinose to L-ribulose in vivo. This enzyme is also of commercial interest as it catalyzes the conversion of D-galactose to D-tagatose in vitro. The crystal structure of ECAI was solved and refined at 2.6 Angstroms resolution. The subunit structure of ECAI is organized into three domains: an N-terminal, a central and a C-terminal domain. It forms a crystallographic trimeric architecture in the asymmetric unit. Packing within the crystal suggests the idea that ECAI can form a hexameric assembly. Previous electron microscopic and biochemical studies supports that ECAI is hexameric in solution. A comparison with other known structures reveals that ECAI adopts a protein fold most similar to E. coli fucose isomerase (ECFI) despite very low sequence identity 9.7%. The structural similarity between ECAI and ECFI with regard to number of domains, overall fold, biological assembly, and active site architecture strongly suggests that the enzymes have functional similarities. Further, the crystal structure of ECAI forms a basis for identifying molecular determinants responsible for isomerization of arabinose to ribulose in vivo and galactose to tagatose in vitro.

Intrinsic flux pinning mechanisms in different thickness MgB2 films

Directory of Open Access Journals (Sweden)

C. Yang

2017-03-01

Full Text Available MgB2 films in four thickness (60 nm, 200nm, 600nm and 1μm have been fabricated by hybrid physical–chemical vapor deposition technique (HPCVD. By measuring the magnetization hysteresis loops and the resistivity, we have obtained the transport and magnetic properties of the four films. After that, the pinning mechanisms in them were discussed. Comparing the pinning behaviors in these ultrathin films, thin films and thick films, it was found that there exist different pinning types in MgB2 films of different thickness. In combination with the study of the surface morphology, cross-section and XRD results, we concluded that MgB2 films had different growth modes in different growth stages. For thin films, films grew along c axis, and grain boundaries acted as surface pinning. While for thick films, films grew along c axis at first, and then changed to a-b axis growth. As a result, the a-b axis grains acted as strong volume pinning.

Pinning impulsive synchronization of stochastic delayed coupled networks

International Nuclear Information System (INIS)

Tang Yang; Fang Jian-An; Wong W K; Miao Qing-Ying

2011-01-01

In this paper, the pinning synchronization problem of stochastic delayed complex network (SDCN) is investigated by using a novel hybrid pinning controller. The proposed hybrid pinning controller is composed of adaptive controller and impulsive controller, where the two controllers are both added to a fraction of nodes in the network. Using the Lyapunov stability theory and the novel hybrid pinning controller, some sufficient conditions are derived for the exponential synchronization of such dynamical networks in mean square. Two numerical simulation examples are provided to verify the effectiveness of the proposed approach. The simulation results show that the proposed control scheme has a fast convergence rate compared with the conventional adaptive pinning method. (general)

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

The Effect of Insertion Technique on Temperatures for Standard and Self-Drilling External Fixation Pins.

Science.gov (United States)

Manoogian, Sarah; Lee, Adam K; Widmaier, James C

2017-08-01

No studies have assessed the effects of parameters associated with insertion temperature in modern self-drilling external fixation pins. The current study assessed how varying the presence of irrigation, insertion speed, and force impacted the insertion temperatures of 2 types of standard and self-drilling external fixation half pins. Seventy tests were conducted with 10 trials for 4 conditions on self-drilling pins, and 3 conditions for standard pins. Each test used a thermocouple inside the pin to measure temperature rise during insertion. Adding irrigation to the standard pin insertion significantly lowered the maximum temperature (P drilling pin tests dropped average rise in temperature from 151.3 ± 21.6°C to 124.1 ± 15.3°C (P = 0.005). When the self-drilling pin insertion was decreased considerably from 360 to 60 rpm, the temperature decreased significantly from 151.3 ± 21.6°C to 109.6 ± 14.0°C (P drilling pin temperature increase was not significant. The standard pin had lower peak temperatures than the self-drilling pin for all conditions. Moreover, slowing down the insertion speed and adding irrigation helped mitigate the temperature increase of both pin types during insertion.

Identification and characterization of a novel L-arabinose isomerase from Anoxybacillus flavithermus useful in D-tagatose production.

Science.gov (United States)

Li, Yanjun; Zhu, Yueming; Liu, Anjun; Sun, Yuanxia

2011-05-01

D-Tagatose is a highly functional rare ketohexose and many attempts have been made to convert D-galactose into the valuable D-tagatose using L-arabinose isomerase (L-AI). In this study, a thermophilic strain possessing L-AI gene was isolated from hot spring sludge and identified as Anoxybacillus flavithermus based on its physio-biochemical characterization and phylogenetic analysis of its 16s rRNA gene. Furthermore, the gene encoding L-AI from A. flavithermus (AFAI) was cloned and expressed at a high level in E. coli BL21(DE3). L-AI had a molecular weight of 55,876 Da, an optimum pH of 10.5 and temperature of 95°C. The results showed that the conversion equilibrium shifted to more D-tagatose from D-galactose by raising the reaction temperatures and adding borate. A 60% conversion of D-galactose to D-tagatose was observed at an isomerization temperature of 95°C with borate. The catalytic efficiency (k (cat) /K (m)) for D-galactose with borate was 9.47 mM(-1) min(-1), twice as much as that without borate. Our results indicate that AFAI is a novel hyperthermophilic and alkaliphilic isomerase with a higher catalytic efficiency for D-galactose, suggesting its great potential for producing D-tagatose.

Theoretical interpretation of SCARABEE single pin in-pile boiling experiments

International Nuclear Information System (INIS)

Struwe, D.; Bottoni, M.; Fries, W.; Elbel, H.; Angerer, G.

1977-01-01

In the framework of LMFBR safety analysis a theoretical interpretation of some of the most representative of the single pin experiments of the in-pile SCARABEE project has been performed from both viewpoints of thermohydraulic and fuel behaviour using the computer codes CAPRI-2 and SATURN-1. The analysis is aimed at investigating the pin behavior from the preirradiation history, through the observed sequence of events following a coolant mass flow reduction from boiling inception up to pin breakdown. A comparison of theoretical results with experimentally recorded data has allowed a deeper insight into the peculiar features of the experiments and enabled a valuable code verification. (Auth.)

Pin-wise Reactor Analysis Based on the Generalized Equivalence Theory

Energy Technology Data Exchange (ETDEWEB)

Yu, Hwan Yeal; Heo, Woong; Kim, Yong Hee [KAIST, Daejeon (Korea, Republic of)

2016-05-15

In this paper, a pin-wise reactor analysis is performed based on the generalized equivalence theory. From the conventional fuel assembly lattice calculations, pin-wise 2-group cross sections and pin DFs are generated. Based on the numerical results on a small PWR benchmark, it is observed that the pin-wise core analysis provide quite accurate prediction on the effective multiplication factor and the peak pin power error is bounded by about 3% in peripheral fuel assemblies facing the baffle-reflector. Also, it was found that relatively large pin power errors occur along the interface between clearly different fuel assemblies. It is expected that the GET-based pin-by-pin core calculation can be further developed as an advanced method for reactor analysis via improving the group constants and discontinuity factors. Recently, high-fidelity multi-dimensional analysis tools are gaining more attention because of their accurate prediction of local parameters for core design and safety assessment. In terms of accuracy, direct whole-core transport is quite promising. However, it is clear that it is still very costly in terms of the computing time and memory requirements. Another possible solution is the pin-by-pin core analysis in which only small fuel pins are homogenized and the 3-D core analysis is still performed using a low-order operator such as the diffusion theory. In this paper, a pin-by-pin core analysis is performed using the hybrid CMFD (HCMFD) method. Hybrid CMFD is a new global-local iteration method that has been developed for efficient parallel calculation of pinby-pin heterogeneous core analysis. For the HCMFD method, the one-node CMFD scheme is combined with a local two-node CMFD method in a non-linear way. Since the SPH method is iterative and SPH factors are not direction dependent, it is clear that SPH method takes more computing cost and cannot take into account the different heterogeneity and transport effects at each pin interface. Unlike the SPH

Pinning impulsive control algorithms for complex network

Energy Technology Data Exchange (ETDEWEB)

Sun, Wen [School of Information and Mathematics, Yangtze University, Jingzhou 434023 (China); Lü, Jinhu [Academy of Mathematics and Systems Science, Chinese Academy of Sciences, Beijing 100190 (China); Chen, Shihua [College of Mathematics and Statistics, Wuhan University, Wuhan 430072 (China); Yu, Xinghuo [School of Electrical and Computer Engineering, RMIT University, Melbourne VIC 3001 (Australia)

2014-03-15

In this paper, we further investigate the synchronization of complex dynamical network via pinning control in which a selection of nodes are controlled at discrete times. Different from most existing work, the pinning control algorithms utilize only the impulsive signals at discrete time instants, which may greatly improve the communication channel efficiency and reduce control cost. Two classes of algorithms are designed, one for strongly connected complex network and another for non-strongly connected complex network. It is suggested that in the strongly connected network with suitable coupling strength, a single controller at any one of the network's nodes can always pin the network to its homogeneous solution. In the non-strongly connected case, the location and minimum number of nodes needed to pin the network are determined by the Frobenius normal form of the coupling matrix. In addition, the coupling matrix is not necessarily symmetric or irreducible. Illustrative examples are then given to validate the proposed pinning impulsive control algorithms.

Pinning impulsive control algorithms for complex network

International Nuclear Information System (INIS)

Sun, Wen; Lü, Jinhu; Chen, Shihua; Yu, Xinghuo

2014-01-01

In this paper, we further investigate the synchronization of complex dynamical network via pinning control in which a selection of nodes are controlled at discrete times. Different from most existing work, the pinning control algorithms utilize only the impulsive signals at discrete time instants, which may greatly improve the communication channel efficiency and reduce control cost. Two classes of algorithms are designed, one for strongly connected complex network and another for non-strongly connected complex network. It is suggested that in the strongly connected network with suitable coupling strength, a single controller at any one of the network's nodes can always pin the network to its homogeneous solution. In the non-strongly connected case, the location and minimum number of nodes needed to pin the network are determined by the Frobenius normal form of the coupling matrix. In addition, the coupling matrix is not necessarily symmetric or irreducible. Illustrative examples are then given to validate the proposed pinning impulsive control algorithms

Overexpressed cyclophilin B suppresses apoptosis associated with ROS and Ca2+ homeostasis after ER stress.

Science.gov (United States)

Kim, Jinhwan; Choi, Tae Gyu; Ding, Yan; Kim, Yeonghwan; Ha, Kwon Soo; Lee, Kyung Ho; Kang, Insug; Ha, Joohun; Kaufman, Randal J; Lee, Jinhwa; Choe, Wonchae; Kim, Sung Soo

2008-11-01

Prolonged accumulation of misfolded proteins in the endoplasmic reticulum (ER) results in ER stress-mediated apoptosis. Cyclophilins are protein chaperones that accelerate the rate of protein folding through their peptidyl-prolyl cis-trans isomerase (PPIase) activity. In this study, we demonstrated that ER stress activates the expression of the ER-localized cyclophilin B (CypB) gene through a novel ER stress response element. Overexpression of wild-type CypB attenuated ER stress-induced cell death, whereas overexpression of an isomerase activity-defective mutant, CypB/R62A, not only increased Ca(2+) leakage from the ER and ROS generation, but also decreased mitochondrial membrane potential, resulting in cell death following exposure to ER stress-inducing agents. siRNA-mediated inhibition of CypB expression rendered cells more vulnerable to ER stress. Finally, CypB interacted with the ER stress-related chaperones, Bip and Grp94. Taken together, we concluded that CypB performs a crucial function in protecting cells against ER stress via its PPIase activity.

Kinetic analysis of the mechanism and specificity of protein-disulfide isomerase using fluorescence-quenched peptides

DEFF Research Database (Denmark)

Westphal, V; Spetzler, J C; Meldal, M

1998-01-01

Protein-disulfide isomerase (PDI) is an abundant folding catalyst in the endoplasmic reticulum of eukaryotic cells. PDI introduces disulfide bonds into newly synthesized proteins and catalyzes disulfide bond isomerizations. We have synthesized a library of disulfide-linked fluorescence......-quenched peptides, individually linked to resin beads, for two purposes: 1) to probe PDI specificity, and 2) to identify simple, sensitive peptide substrates of PDI. Using this library, beads that became rapidly fluorescent by reduction by human PDI were selected. Amino acid sequencing of the bead-linked peptides...

Fabrication and post-irradiation examination of a zircaloy-2 clad UO2-1.5 wt% PuO2 fuel pin irradiated in PWL, CIRUS

International Nuclear Information System (INIS)

Sah, D.N.; Sahoo, K.C.; Chatterjee, S.; Majumdar, S.; Kamath, H.S.; Ramachandran, R.; Bahl, J.K.; Purushottam, D.S.C.; Ramakumar, M.S.; Sivaramakrishnan, K.S.; Roy, P.R.

1977-01-01

A zircaloy-2 clad UO 2 -1.5 wt% PuO 2 fuel pin was fabricated at the Radiometallurgy Section of the Bhabha Atomic Research Centre, Bombay, for irradiation in the pressurised water loop in CIRUS. Requisite development work related to powder conditioning, blending, pressing and sintering parameters was carried out to meet the exacting fuel pellet specifications of CANDU fuel. The fuel pin ruptured while being irradiated in the pressurised water loop in CIRUS, after experiencing a low burn-up of 507 MWD/MTM and was subsequently examined at the Radiometallurgy Hot Cells Facility. The results showed that internal clad hydriding led to primary failure of the fuel pin. Subsequent ingress of the coolant water caused excessive swelling of the thermal insulating magnesia pellets located at the ends of the fuel column. The swelling of magnesia pellets caused severe rupturing of the fuel pin at the two ends. The delayed rupturing of the fuel pin at the upper end, caused the fuel column to be displaced downwards by 5.85mm. (author)

Novel pinning control strategies for synchronisation of complex networks with nonlinear coupling dynamics

International Nuclear Information System (INIS)

Zhao-Bing, Liu; Hua-Guang, Zhang; Qiu-Ye, Sun

2010-01-01

This paper considers the global stability of controlling an uncertain complex network to a homogeneous trajectory of the uncoupled system by a local pinning control strategy. Several sufficient conditions are derived to guarantee the network synchronisation by investigating the relationship among pinning synchronisation, network topology, and coupling strength. Also, some fundamental and yet challenging problems in the pinning control of complex networks are discussed: (1) what nodes should be selected as pinned candidates? (2) How many nodes are needed to be pinned for a fixed coupling strength? Furthermore, an adaptive pinning control scheme is developed. In order to achieve synchronisation of an uncertain complex network, the adaptive tuning strategy of either the coupling strength or the control gain is utilised. As an illustrative example, a network with the Lorenz system as node self-dynamics is simulated to verify the efficacy of theoretical results. (general)

Production of D-tagatose at high temperatures using immobilized Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana.

Science.gov (United States)

Hong, Young-Ho; Lee, Dong-Woo; Lee, Sang-Jae; Choe, Eun-Ah; Kim, Seong-Bo; Lee, Yoon-Hee; Cheigh, Chan-Ick; Pyun, Yu-Ryang

2007-04-01

Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana (TNAI) were immobilized in calcium alginate beads. The resulting cell reactor (2.4 U, t (1/2) = 43 days at 70 degrees C) in a continuous recycling mode at 70 degrees C produced 49 and 38 g D-tagatose/l from 180 and 90 g D-galactose/l, respectively, within 12 h.

Cluster synchronization for directed community networks via pinning partial schemes

International Nuclear Information System (INIS)

Hu Cheng; Jiang Haijun

2012-01-01

Highlights: ► Cluster synchronization for directed community networks is proposed by pinning partial schemes. ► Each community is considered as a whole. ► Several novel pinning criteria are derived based on the information of communities. ► A numerical example with simulation is provided. - Abstract: In this paper, we focus on driving a class of directed networks to achieve cluster synchronization by pinning schemes. The desired cluster synchronization states are no longer decoupled orbits but a set of un-decoupled trajectories. Each community is considered as a whole and the synchronization criteria are derived based on the information of communities. Several pinning schemes including feedback control and adaptive strategy are proposed to select controlled communities by analyzing the information of each community such as indegrees and outdegrees. In all, this paper answers several challenging problems in pinning control of directed community networks: (1) What communities should be chosen as controlled candidates? (2) How many communities are needed to be controlled? (3) How large should the control gains be used in a given community network to achieve cluster synchronization? Finally, an example with numerical simulations is given to demonstrate the effectiveness of the theoretical results.

Vortex pinning by point defect in superconductors

International Nuclear Information System (INIS)

Liao Hongyin; Zhou Shiping; Du Haochen

2003-01-01

We apply the periodic time-dependent Ginzburg-Landau model to study vortex distribution in type-II superconductors with a point-like defect and square pinning array. A defect site will pin vortices, and a periodic pinning array with right geometric parameters, which can be any form designed in advance, shapes the vortex pattern as external magnetic field varies. The maximum length over which an attractive interaction between a pinning centre and a vortex extends is estimated to be about 6.0ξ. We also derive spatial distribution expressions for the order parameter, vector potential, magnetic field and supercurrent induced by a point defect. Theoretical results and numerical simulations are compared with each other and they are consistent

1 GSPS digitizer based on the FPGA Mezzanine Card (FMC) standard with low-count pin connector.

CERN Document Server

Vasilyev, Mikhail

2015-01-01

Under the scope of a CERN summer student project, the schematic for ADC based on FMC mezzanine card with 1 GSPS sampling rate and 8 bit resolution was developed. The mezzanine is fully compatible with the standard: FPGA Mezzanine Card (FMC) [1]. A low-pin count connector was used to connect the mezzanine with the “carrier”. The carrier was an Open Hardware project: Simple PCIe FMC carrier (SPEC).

Coexpression of β-D-galactosidase and L-arabinose isomerase in the production of D-tagatose: a functional sweetener.

Science.gov (United States)

Zhan, Yijing; Xu, Zheng; Li, Sha; Liu, Xiaoliu; Xu, Lu; Feng, Xiaohai; Xu, Hong

2014-03-19

The functional sweetener, d-tagatose, is commonly transformed from galactose by l-arabinose isomerase. To make use of a much cheaper starting material, lactose, hydrolization, and isomerization are required to take place collaboratively. Therefore, a single-step method involving β-d-galactosidase was explored for d-tagatose production. The two vital genes, β-d-galactosidase gene (lacZ) and l-arabinose isomerase mutant gene (araA') were extracted separately from Escherichia coli strains and incorporated into E. coli simultaneously. This gave us E. coli-ZY, a recombinant producing strain capable of coexpressing the two key enzymes. The resulted cells exhibited maximum d-tagatose producing activity at 34 °C and pH 6.5 and in the presence of borate, 10 mM Fe(2+), and 1 mM Mn(2+). Further monitoring showed that the recombinant cells could hydrolyze more than 95% lactose and convert 43% d-galactose into d-tagatose. This research has verified the feasibility of single-step d-tagatose fermentation, thereby laying down the foundation for industrial usage of lactose.

Inscuteable Regulates the Pins-Mud Spindle Orientation Pathway

Science.gov (United States)

Mauser, Jonathon F.; Prehoda, Kenneth E.

2012-01-01

During asymmetric cell division, alignment of the mitotic spindle with the cell polarity axis ensures that the cleavage furrow separates fate determinants into distinct daughter cells. The protein Inscuteable (Insc) is thought to link cell polarity and spindle positioning in diverse systems by binding the polarity protein Bazooka (Baz; aka Par-3) and the spindle orienting protein Partner of Inscuteable (Pins; mPins or LGN in mammals). Here we investigate the mechanism of spindle orientation by the Insc-Pins complex. Previously, we defined two Pins spindle orientation pathways: a complex with Mushroom body defect (Mud; NuMA in mammals) is required for full activity, whereas binding to Discs large (Dlg) is sufficient for partial activity. In the current study, we have examined the role of Inscuteable in mediating downstream Pins-mediated spindle orientation pathways. We find that the Insc-Pins complex requires Gαi for partial activity and that the complex specifically recruits Dlg but not Mud. In vitro competition experiments revealed that Insc and Mud compete for binding to the Pins TPR motifs, while Dlg can form a ternary complex with Insc-Pins. Our results suggest that Insc does not passively couple polarity and spindle orientation but preferentially inhibits the Mud pathway, while allowing the Dlg pathway to remain active. Insc-regulated complex assembly may ensure that the spindle is attached to the cortex (via Dlg) before activation of spindle pulling forces by Dynein/Dynactin (via Mud). PMID:22253744

Establishing the need for an engineering standard for agricultural hitch pins.

Science.gov (United States)

Deboy, G R; Knapp, W M; Field, W E; Krutz, G W; Corum, C L

2012-04-01

Documented incidents have occurred in which failure or unintentional disengagement of agricultural hitch pins has contributed to property damage and personal injury. An examination of current hitch pin use on a convenience sample of farm operations in Indiana revealed a variety of non-standard, worn and damaged, and inappropriately sized hitch pins in use. Informal interviews with the farm operators confirmed that hitch pin misuse, failure, or disengagement is a relatively widespread problem that remains largely unaddressed. On-site observations also suggested a low use of hitch pin retaining devices or safety chains. A review of prior research revealed that little attention has been given to this problem, and currently no documentation allows for an estimate of the frequency or severity of losses associated with hitch pin misuse, failure, or disengagement. No specific engineering standards were found that directly applied to the design, appropriate selection, or loading capacity of agricultural hitch pins. Major suppliers of replacement hitch pins currently provide little or no information on matching hitch pin size to intended applications, and most replacement hitch pins examined were of foreign origin, with the overwhelming majority imported from China or India. These replacement hitch pins provided no specifications other than diameter, length, and, in some cases, labeling that indicated that the pins had been "heat treated. " Testing of a sample of 11 commercially available replacement hitch pins found variation along the length of the pin shaft and between individual pins in surface hardness, a potential predictor of pin failure. Examination of 17 commercially available replacement pins also revealed a variety of identifiers used to describe pin composition and fabrication methods, e.g., "heat treated." None of the pins examined provided any specifications on loading capacity. It was therefore concluded that there is a need to develop an agricultural hitch

Vortex pinning in artificially layered Ba(Fe,Co)2As2 film

Science.gov (United States)

Oh, M. J.; Lee, Jongmin; Seo, Sehun; Yoon, Sejun; Seo, M. S.; Park, S. Y.; Kim, Ho-Sup; Ha, Dong-Woo; Lee, Sanghan; Jo, Youn Jung

2018-06-01

Static high critical current densities (Jc) > 1 MA/cm2 with magnetic field parallel or perpendicular to c-axis were realized in Co-doped/undoped multilayerd BaFe2As2 films. We made a current bridge by FIB to allow precise measurements, and confirmed that the boundary quality using FIB was considerably better than the quality achieved using a laser. The presence of a high in-plane Jc suggested the existence of c-axis correlated vortex pinning centers. To clarify the relationship between the Jc performance and superstructures, we investigated the magnetic flux pinning mechanism using scaling theory of the volume pinning force Fp(H). The Jc(H) curves, Fp/Fp,max vs. h = H/Hirr curves, and parameters p and q depended on the characteristics of the flux pinning mechanism. It was found that the dominant pinning mechanism of Co-doped/undoped multilayerd BaFe2As2 films was Δl-pinning and the inserted undoped BaFe2As2 layers remained non-superconducting. The dominant pin geometry varied when the magnetic field direction changed. It was concluded that the artificially layered BaFe2As2 film is a 3-D superconductor due to its long correlation length compared to the thickness of the non-superconducting layer.

The acid-tolerant L-arabinose isomerase from the mesophilic Shewanella sp. ANA-3 is highly active at low temperatures

Science.gov (United States)

2011-01-01

Background L-arabinose isomerases catalyse the isomerization of L-arabinose into L-ribulose at insight biological systems. At industrial scale of this enzyme is used for the bioconversion of D-galactose into D-tagatose which has many applications in pharmaceutical and agro-food industries. The isomerization reaction is thermodynamically equilibrated, and therefore the bioconversion rates is shifted towards tagatose when the temperature is increased. Moreover, to prevent secondary reactions it will be of interest to operate at low pH. The profitability of this D-tagatose production process is mainly related to the use of lactose as cheaper raw material. In many dairy products it will be interesting to produce D-tagatose during storage. This requires an efficient L-arabinose isomerase acting at low temperature and pH values. Results The gene encoding the L-arabinose isomerase from Shewanella sp. ANA-3 was cloned and overexpressed in Escherichia coli. The purified protein has a tetrameric arrangement composed by four identical 55 kDa subunits. The biochemical characterization of this enzyme showed that it was distinguishable by its maximal activity at low temperatures comprised between 15-35°C. Interestingly, this biocatalyst preserves more than 85% of its activity in a broad range of temperatures from 4.0 to 45°C. Shewanella sp. ANA-3 L-arabinose isomerase was also optimally active at pH 5.5-6.5 and maintained over 80% of its activity at large pH values from 4.0 to 8.5. Furthermore, this enzyme exhibited a weak requirement for metallic ions for its activity evaluated at 0.6 mM Mn2+. Stability studies showed that this protein is highly stable mainly at low temperature and pH values. Remarkably, T268K mutation clearly enhances the enzyme stability at low pH values. Use of this L-arabinose isomerase for D-tagatose production allows the achievement of attractive bioconversion rates of 16% at 4°C and 34% at 35°C. Conclusions Here we reported the purification and the

The acid-tolerant L-arabinose isomerase from the mesophilic Shewanella sp. ANA-3 is highly active at low temperatures

Directory of Open Access Journals (Sweden)

Rhimi Moez

2011-11-01

Full Text Available Abstract Background L-arabinose isomerases catalyse the isomerization of L-arabinose into L-ribulose at insight biological systems. At industrial scale of this enzyme is used for the bioconversion of D-galactose into D-tagatose which has many applications in pharmaceutical and agro-food industries. The isomerization reaction is thermodynamically equilibrated, and therefore the bioconversion rates is shifted towards tagatose when the temperature is increased. Moreover, to prevent secondary reactions it will be of interest to operate at low pH. The profitability of this D-tagatose production process is mainly related to the use of lactose as cheaper raw material. In many dairy products it will be interesting to produce D-tagatose during storage. This requires an efficient L-arabinose isomerase acting at low temperature and pH values. Results The gene encoding the L-arabinose isomerase from Shewanella sp. ANA-3 was cloned and overexpressed in Escherichia coli. The purified protein has a tetrameric arrangement composed by four identical 55 kDa subunits. The biochemical characterization of this enzyme showed that it was distinguishable by its maximal activity at low temperatures comprised between 15-35°C. Interestingly, this biocatalyst preserves more than 85% of its activity in a broad range of temperatures from 4.0 to 45°C. Shewanella sp. ANA-3 L-arabinose isomerase was also optimally active at pH 5.5-6.5 and maintained over 80% of its activity at large pH values from 4.0 to 8.5. Furthermore, this enzyme exhibited a weak requirement for metallic ions for its activity evaluated at 0.6 mM Mn2+. Stability studies showed that this protein is highly stable mainly at low temperature and pH values. Remarkably, T268K mutation clearly enhances the enzyme stability at low pH values. Use of this L-arabinose isomerase for D-tagatose production allows the achievement of attractive bioconversion rates of 16% at 4°C and 34% at 35°C. Conclusions Here we

Ultrasonic inspections of fuel alignment pins

International Nuclear Information System (INIS)

Rathgeb, W.; Schmid, R.

1994-01-01

As a remedy to the practical problem of defects in fuel alignment pins made of Inconel X750, an inspection technique has been developed which fully meets the requirements of detecting defects. The newly used fuel alignment pins made of austenite are easy to test and therefore satisfy the necessity of further inspections.For the fuel alignment pins of the upper core structure a safe and fast inspection technique was made available. The inspection sensitivity is high and it is possible to give quantitative directions concerning defect orientation and depth. After the required inspections had been concluded in 1989, a total of 18 inspections were carried out in various national and international nuclear power plants in the following years. During this time more than 6000 fuel alignment pines were examined.For the fuel alignment pins the inspection technique provided could increase the understanding of the defect process. This technique contributed to the development of an adaptive and economical repair strategy. ((orig.))

Design and Operation of 3-Pin FTL HVAC System

International Nuclear Information System (INIS)

Chi, D. Y.; Sim, B. S.; Park, S. K.; Park, K. N.; Lee, J. M.; Ahn, S. H.; Lee, C. Y.; Kim, Y. J.

2005-01-01

According to the increasing demand for irradiation tests to develop new fuels, the 3-Pin FTL(Fuel Test Loop for 3 pin test fuel) facility has now been under design to conduct in-core fuel performance tests at the operating conditions, which will be installed at HANARO. The HVAC system of the FTL will be dependent on that of the HANARO. The FTL has three equipments rooms, which are the room 1, room 2 and the control room. The high pressure and high temperature equipments will be installed in the room 1. The atmosphere of the room 1 shall be maintained under the designed condition. This paper describes the design of the FTL HVAC system in the room 1

Cell homogenization methods for pin-by-pin core calculations tested in slab geometry

International Nuclear Information System (INIS)

Yamamoto, Akio; Kitamura, Yasunori; Yamane, Yoshihiro

2004-01-01

In this paper, performances of spatial homogenization methods for fuel or non-fuel cells are compared in slab geometry in order to facilitate pin-by-pin core calculations. Since the spatial homogenization methods were mainly developed for fuel assemblies, systematic study of their performance for the cell-level homogenization has not been carried out. Importance of cell-level homogenization is recently increasing since the pin-by-pin mesh core calculation in actual three-dimensional geometry, which is less approximate approach than current advanced nodal method, is getting feasible. Four homogenization methods were investigated in this paper; the flux-volume weighting, the generalized equivalence theory, the superhomogenization (SPH) method and the nonlinear iteration method. The last one, the nonlinear iteration method, was tested as the homogenization method for the first time. The calculations were carried out in simplified colorset assembly configurations of PWR, which are simulated by slab geometries, and homogenization performances were evaluated through comparison with the reference cell-heterogeneous calculations. The calculation results revealed that the generalized equivalence theory showed best performance. Though the nonlinear iteration method can significantly reduce homogenization error, its performance was not as good as that of the generalized equivalence theory. Through comparison of the results obtained by the generalized equivalence theory and the superhomogenization method, important byproduct was obtained; deficiency of the current superhomogenization method, which could be improved by incorporating the 'cell-level discontinuity factor between assemblies', was clarified

Pinning Synchronization of Switched Complex Dynamical Networks

Directory of Open Access Journals (Sweden)

Liming Du

2015-01-01

Full Text Available Network topology and node dynamics play a key role in forming synchronization of complex networks. Unfortunately there is no effective synchronization criterion for pinning synchronization of complex dynamical networks with switching topology. In this paper, pinning synchronization of complex dynamical networks with switching topology is studied. Two basic problems are considered: one is pinning synchronization of switched complex networks under arbitrary switching; the other is pinning synchronization of switched complex networks by design of switching when synchronization cannot achieved by using any individual connection topology alone. For the two problems, common Lyapunov function method and single Lyapunov function method are used respectively, some global synchronization criteria are proposed and the designed switching law is given. Finally, simulation results verify the validity of the results.

Breached-pin testing in the US

International Nuclear Information System (INIS)

Mahagin, D.E.; Lambert, J.D.B.

1981-04-01

Experience gained at EBR-II by the late 1970's from a significant number of failures in experimental fuel-pin irradiations forms the basis of a program directed towards the characterization of breached pins. The questions to be answered and the issues raised by further testing are discussed

Mannose Phosphate Isomerase Isoenzymes in Plutella xylostella Support Common Genetic Bases of Resistance to Bacillus thuringiensis Toxins in Lepidopteran Species

OpenAIRE

Herrero, Salvador; Ferré, Juan; Escriche, Baltasar

2001-01-01

A strong correlation between two mannose phosphate isomerase (MPI) isoenzymes and resistance to Cry1A toxins from Bacillus thuringiensis has been found in a Plutella xylostella population. MPI linkage to Cry1A resistance had previously been reported for a Heliothis virescens population. The fact that the two populations share similar biochemical, genetic, and cross-resistance profiles of resistance suggests the occurrence of homologous resistance loci in both species.

Is magnetic pinning a dominant mechanism in Nb-Ti

International Nuclear Information System (INIS)

Cooley, L.D.; Lee, P.J.; Larbalestier, D.C.

1991-01-01

In this paper, the authors compare the pinning behavior of an artificial pinning center (APC) composite and a nanometer-filament Nb 46.5 wt% Ti composite to that of a conventional Nb 48 wt% Ti composite. The microstructure of the APC composite resembles the conventional composite, where ribbons of normal metal form the pinning centers, whereas the nanometer-filament composite has no internal normal metal but pins instead at the filament surface. The APC composite exhibits much stronger pinning relative to B c 2 than the conventional composite (21.4 GN/m 3 , 7 T vs. 18.9 GN/m 3 , 11 T), which is possibly due to the increased amount of pinning center (50 vol.% vs. 25 vol.%), however the proximity effect reduces the B c 2 unfavorably

Velocity distribution measurement in wire-spaced fuel pin bundle

International Nuclear Information System (INIS)

Mizuta, Hiroshi; Ohtake, Toshihide; Uruwashi, Shinichi; Takahashi, Keiichi

1974-01-01

Flow distribution measurement was made in the subchannels of a pin bundle in air flow. The present paper is interim because the target of this work is the decision of temperature of the pin surface in contact with wire spacers. The wire-spaced fuel pin bundle used for the experiment consists of 37 simulated fuel pins of stainless steel tubes, 3000 mm in length and 31.6 mm in diameter, which are wound spirally with 6 mm stainless steel wire. The bundle is wrapped with a hexagonal tube, 3500 mm in length and 293 mm in flat-to-flat distance. The bundle is fixed with knock-bar at the entrance of air flow in the hexagonal tube. The pitch of pins in the bundle is 37.6 mm (P/D=1.19) and the wrapping pitch of wire is 1100 mm (H/D=34.8). A pair of arrow-type 5-hole Pitot tubes are used to measure the flow velocity and the direction of air flow in the pin bundle. The measurement of flow distribution was made with the conditions of air flow rate of 0.33 m 3 /sec, air temperature of 45 0 C, and average Reynolds number of 15100 (average air velocity of 20.6 m/sec.). It was found that circular flow existed in the down stream of wire spacers, that axial flow velocity was slower in the subchannels, which contained wire spacers, than in those not affected by the wire, and that the flow angle to the axial velocity at the boundary of subchannels was two thirds smaller than wire wrapping angle. (Tai, I.)

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.

Evaluation of bundle duct interaction by out-of-pile compression test of FBR fuel pin bundles

Energy Technology Data Exchange (ETDEWEB)

Tanaka, Kosuke; Yamamoto, Yuji; Nagamine, Tsuyoshi; Maeda, Koji [Japan Nuclear Cycle Development Inst., Oarai, Ibaraki (Japan). Oarai Engineering Center

2001-06-01

Bundle duct interaction (BDI) caused by expansion of fuel pin bundle is a main factor to limit the fuel lifetime. Therefore, it is important for the design of fast reactor fuel assembly to understand the fuel pin deformation behavior under BDI condition. In order to understand the fuel pin deformation behavior under BDI condition, out-of-pile compression tests were conducted for FBR fuel pin bundle by use of X-ray CT equipment. In these compression tests, two kinds of fuel pin bundles were conducted. One was the fuel pin bundle with the short wire-pitch and the other was the fuel pin bundle with the short wire-pitch and large diameter claddings. The general discussions were also performed based on the results of out-of-pile compression tests obtained by use of X-ray CT equipment in the previous work. Following results were obtained. 1) The occurrence of the pin-to-duct contact depends on the wire-pitch. In the fuel pin bundle with large wire-pitch, the pin-to-duct contact occurred at the early stage of BDI. The reason of this result is due to the low bowing rigidity of the fuel pins with long wire-pitch. 2) The value of the ovalation stiffness strongly depends on the geometry of cladding (diameter, thickness) and especially on wire-pitch. This result in this work revealed that the occurrence of the pin-to-duct contact depends on the value of the ovalation stiffness. 3) The occurrence of wire dispersion and dispersive displacement of pins depends on the wire-pitch strongly. In the fuel pin bundle with the long wire-pitch, the occurrence of the above-mentioned suppression mechanism to BDI is remarkable. 4) The suppression mechanism to BDI of the fuel pin bundle with the long wire-pitch is elastic oval deformation of cladding, wire dispersion and dispersive displacement of pins. On the other hand, the elastic and plastic oval deformation of cladding is the major suppression mechanism to BDI in the fuel pin bundle with the short wire-pitch. 5) The appearance of

Control analysis of the role of triosephosphate isomerase in glucose metabolism in Lactococcus lactis

DEFF Research Database (Denmark)

Solem, Christian; Købmann, Brian Jensen; Jensen, Peter Ruhdal

2008-01-01

Triosephosphate isomerase (TPI), which catalyses the conversion of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (G3P), was studied for its control on glycolysis and mixed acid production in L. lactis subspecies lactis IL1403 and L. lactis subspecies cremoris MG1363. Strains...... metabolites glucose-6-phosphate, fructose-1,6-bisphosphate and DHAP in the IL1403 derivatives were essentially unchanged for TPI activities from 26% to 225%. At a TPI activity of 3%, the level of DHAP increased four times. The finding that an increased level of DHAP coincides with an increase in formate...

End-threaded intramedullary positive profile screw ended self-tapping pin (Admit pin) - A cost-effective novel implant for fixing canine long bone fractures.

Science.gov (United States)

Chanana, Mitin; Kumar, Adarsh; Tyagi, Som Prakash; Singla, Amit Kumar; Sharma, Arvind; Farooq, Uiase Bin

2018-02-01

The current study was undertaken to evaluate the clinical efficacy of end-threaded intramedullary pinning for management of various long bone fractures in canines. This study was conducted in two phases, managing 25 client-owned dogs presented with different fractures. The technique of application of end-threaded intramedullary pinning in long bone fractures was initially standardized in 6 clinical patients presented with long bone fractures. In this phase, end-threaded pins of different profiles, i.e., positive and negative, were used as the internal fixation technique. On the basis of results obtained from standardization phase, 19 client-owned dogs clinically presented with different fractures were implanted with end-threaded intramedullary positive profile screw ended self-tapping pin in the clinical application phase. The patients, allocated randomly in two groups, when evaluated postoperatively revealed slight pin migration in Group-I (negative profile), which resulted in disruption of callus site causing delayed union in one case and large callus formation in other two cases whereas no pin migration was observed in Group-II (positive profile). Other observations in Group-I was reduced muscle girth and delayed healing time as compared to Group-II. In clinical application, phase 21 st and 42 nd day post-operative radiographic follow-up revealed no pin migration in any of the cases, and there was no bone shortening or fragment collapse in end-threaded intramedullary positive profile screw ended self-tapping pin. The end-threaded intramedullary positive profile screw ended self-tapping pin used for fixation of long bone fractures in canines can resist pin migration, pin breakage, and all loads acting on the bone, i.e., compression, tension, bending, rotation, and shearing to an extent with no post-operative complications.

Happy Pinning

DEFF Research Database (Denmark)

Fausing, Bent

2012-01-01

This is about Pinterest, but with a different approach than usual to social networks. Pinterest is an image site par excellence. The images are as Windows that open outwards and also lets us look inwards and displays the soul and heart, the unintentional or pre-conscious desires. Happy Pinning!...

Suspension scheme for fuel pin

International Nuclear Information System (INIS)

Butts, C.E.; Gray, H.C.

1975-01-01

A description is presented of a nuclear fuel pin suspension arrangement comprising, in combination, a rod; a first beam member connected to said rod at one end; a plurality of parallel-spaced slidable fuel support plates attached to said first beam member, the longitudinal axis of first beam member being perpendicular to the longitudinal axis of each of said fuel support plates, a first coupling means disposed along the length of the first beam member for permitting slidable fuel support plates parallel movement with respect to the longitudinal axis of said first beam member, a second coupling means located at one end of each of slidable fuel plates for slidably engaging first coupling means of first beam member, a second beam member connected to the other end of each of parallel-spaced slidable fuel support plates and providing an extension, second beam member being provided with a third coupling means disposed along the length of second beam member at one end thereof; and a plurality of fuel pins provided with a fourth coupling means located at one end of each fuel pin for slidably engaging third coupling means of second beam member to permit each fuel pin parallel movement with respect to the longitudinal axis of second beam member. (U.S.)

Coevolving MAPK and PID phosphosites indicate an ancient environmental control of PIN auxin transporters in land plants.

Science.gov (United States)

Dory, Magdalena; Hatzimasoura, Elizabeth; Kállai, Brigitta M; Nagy, Szilvia K; Jäger, Katalin; Darula, Zsuzsanna; Nádai, Tímea V; Mészáros, Tamás; López-Juez, Enrique; Barnabás, Beáta; Palme, Klaus; Bögre, László; Ditengou, Franck A; Dóczi, Róbert

2018-01-01

Plant growth flexibly adapts to environmental conditions, implying cross-talk between environmental signalling and developmental regulation. Here, we show that the PIN auxin efflux carrier family possesses three highly conserved putative mitogen-activated protein kinase (MAPK) sites adjacent to the phosphorylation sites of the well-characterised AGC kinase PINOID, which regulates the polar localisation of PINs and directional auxin transport, thereby underpinning organ growth. The conserved sites of PIN1 are phosphorylated in vitro by two environmentally activated MAPKs, MPK4 and MPK6. In contrast to AGC kinases, MAPK-mediated phosphorylation of PIN1 at adjacent sites leads to a partial loss of the plasma membrane localisation of PIN1. MAPK-mediated modulation of PIN trafficking may participate in environmental adjustment of plant growth. © 2017 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

Effects of polybrominated diphenyl ethers (PBDEs) and their derivatives on protein disulfide isomerase activity and growth hormone release of GH3 cells.

Science.gov (United States)

Hashimoto, Shoko; Yoshimura, Hiromi; Okada, Kazushi; Uramaru, Naoto; Sugihara, Kazumi; Kitamura, Shigeyuki; Imaoka, Susumu

2012-03-19

Polybrominated diphenyl ethers (PBDEs) have been used in a variety of consumer products such as flame retardants and recently have been known to be widespread environmental pollutants, which probably affect biological functions of mammalian cells. However, the risk posed by PBDE metabolites has not been clarified. Our previous study suggested that bisphenol A (BPA), an endocrine-disrupting chemical, binds to protein disulfide isomerase (PDI) and inhibits its activity. PDI is an isomerase enzyme in the endoplasmic reticulum and facilitates the formation or cleavage of disulfide bonds. PDI consists of a, b, b', and a' domains and the c region, with the a and a' domains having isomerase active sites. In the present study, we tested the effects of 10 kinds of PBDE compounds and their metabolites on PDI. OH-PBDEs specifically inhibited the isomerase activity of PDI, with 4'-OH-PBDE more effective than 2' (or 2)-OH-PBDEs. 4'-OH-PBDE inhibited the isomerase activity of the b'a'c fragment but not that of ab and a'c, suggesting that the b' domain of PDI is essential for the inhibition by 4'-OH-PBDE. We also investigated the effects of these chemicals on the production of growth hormone (GH) in GH3 cells. In GH3 cells, levels of mRNA and protein of GH stimulated by T(3) were reduced by 4'-OH-PBDE and 4'-MeO-PBDE. The reduction in GH expression caused by these compounds was not changed by the overexpression or knockdown of PDI in GH3 cells, while these manipulations of PDI levels significantly suppressed the expression of GH. These results suggest that the biological effects of PBDEs differed depending on their brominated and hydroxylated positions. © 2011 American Chemical Society

Retractable Pin Tools for the Friction Stir Welding Process

Science.gov (United States)

1998-01-01

Two companies have successfully commercialized a specialized welding tool developed at the Marshall Space Flight Center (MSFC). Friction stir welding uses the high rotational speed of a tool and the resulting frictional heat created from contact to crush, 'stir' together, and forge a bond between two metal alloys. It has had a major drawback, reliance on a single-piece pin tool. The pin is slowly plunged into the joint between two materials to be welded and rotated as high speed. At the end of the weld, the single-piece pin tool is retracted and leaves a 'keyhole,' something which is unacceptable when welding cylindrical objects such as drums, pipes and storage tanks. Another drawback is the requirement for different-length pin tools when welding materials of varying thickness. An engineer at the MSFC helped design an automatic retractable pin tool that uses a computer-controlled motor to automatically retract the pin into the shoulder of the tool at the end of the weld, preventing keyholes. This design allows the pin angle and length to be adjusted for changes in material thickness and results in a smooth hole closure at the end of the weld. Benefits of friction stir welding, using the MSFC retractable pin tool technology, include the following: The ability to weld a wide range of alloys, including previously unweldable and composite materials; provision of twice the fatigue resistance of fusion welds and no keyholes; minimization of material distortion; no creation of hazards such as welding fumes, radiation, high voltage, liquid metals, or arcing; automatic retraction of the pin at the end of the weld; and maintaining full penetration of the pin.

Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analysis of the mannose 6-phosphate isomerase from Salmonella typhimurium

Energy Technology Data Exchange (ETDEWEB)

Gowda, Giri; Sagurthi, Someswar Rao [Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012 (India); Savithri, H. S. [Department of Biochemistry, Indian Institute of Science, Bangalore 560 012 (India); Murthy, M. R. N., E-mail: mrn@mbu.iisc.ernet.in [Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012 (India)

2008-02-01

The cloning, expression, purification, crystallization and preliminary X-ray crystallographic studies of mannose 6-phosphate isomerase from S. typhimurium are reported. Mannose 6-phosphate isomerase (MPI; EC 5.3.1.8) catalyzes the reversible isomerization of d-mannose 6-phosphate (M6P) and d-fructose 6-phosphate (F6P). In the eukaryotes and prokaryotes investigated to date, the enzyme has been reported to play a crucial role in d-mannose metabolism and supply of the activated mannose donor guanosine diphosphate d-mannose (GDP-d-mannose). In the present study, MPI was cloned from Salmonella typhimurium, overexpressed in Escherichia coli and purified using Ni–NTA affinity column chromatography. Purified MPI crystallized in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 36.03, b = 92.2, c = 111.01 Å. A data set extending to 1.66 Å resolution was collected with 98.8% completeness using an image-plate detector system mounted on a rotating-anode X-ray generator. The asymmetric unit of the crystal cell was compatible with the presence of a monomer of MPI. A preliminary structure solution of the enzyme has been obtained by molecular replacement using Candida albicans MPI as the phasing model and the program Phaser. Further refinement and model building are in progress.

Correlation of creep and swelling with fuel pin performance

International Nuclear Information System (INIS)

Jackson, R.J.; Washburn, D.F.; Garner, F.A.; Gilbert, E.R.

1975-09-01

The HEDL PNL-11 experiment described was one in a series of fueled subassemblies irradiated in EBR-II to demonstrate the adequacy of the FFTF fuel pin design. The cladding material, dimensions, and fuel density are prototypic of FFTF. Because neutron flux in EBR-II is lower than in FFTF, the uranium enrichment is higher in these experimental fuel pins, irradiated in EBR-II, than the FFTF enrichment for comparable linear heat rates. Some pertinent oprating conditions for the center fuel pin in this experiment are listed. This 37-pin subassembly represents, at 110,000 MWd/MTM, the highest burnup yet attained by a prototypic FFTF subassembly. Similarly, this is the highest fluence presently attained by prototypic fuel pins. A cladding breach occurred in one fuel pin which is presently being examined. Results are presented and discussed

Material composition – Pinning strength correlation in Nb thin films with focused ion beam-milled washboard nanostructures

International Nuclear Information System (INIS)

Dobrovolskiy, Oleksandr V.; Begun, Evgeniya; Huth, Michael; Shklovskij, Valerij A.

2013-01-01

Highlights: •We fabricated an array of grooves in Nb films by using focused ion beam milling. •We determined the material composition in different areas of the processed films. •We deduced the pinning activation energies from the magneto-resistivity data. •We obtained the material composition – pinning strength correlation in the processed films. -- Abstract: An analysis of the interrelated changes in the material composition and the pinning strength in nanostructured Nb (1 1 0) thin films is presented. The nanopatterns were prepared by focused ion beam milling of an array of uniaxial grooves. They induce a washboard-like pinning potential landscape for vortices in the mixed state. By applying different magnetic fields, the most likely pinning sites along which the flux lines move through the samples have been selected. By this, either the background isotropic pinning of the pristine film or the enhanced isotropic pinning originating from the nanoprocessing has been probed. The enhanced pinning strength in the processed films has been found to correlate with the content of Ga implanted into the films during the nanopatterning

Material composition – Pinning strength correlation in Nb thin films with focused ion beam-milled washboard nanostructures

Energy Technology Data Exchange (ETDEWEB)

Dobrovolskiy, Oleksandr V., E-mail: Dobrovolskiy@Physik.uni-frankfurt.de [Physikalisches Institut, Goethe-Universität, 60438 Frankfurt am Main (Germany); Physical Department, Kharkiv National University, 61077 Kharkiv (Ukraine); Begun, Evgeniya; Huth, Michael [Physikalisches Institut, Goethe-Universität, 60438 Frankfurt am Main (Germany); Shklovskij, Valerij A. [Physical Department, Kharkiv National University, 61077 Kharkiv (Ukraine); Institute for Theoretical Physics, NSC-KIPT, 61108 Kharkiv (Ukraine)

2013-11-15

Highlights: •We fabricated an array of grooves in Nb films by using focused ion beam milling. •We determined the material composition in different areas of the processed films. •We deduced the pinning activation energies from the magneto-resistivity data. •We obtained the material composition – pinning strength correlation in the processed films. -- Abstract: An analysis of the interrelated changes in the material composition and the pinning strength in nanostructured Nb (1 1 0) thin films is presented. The nanopatterns were prepared by focused ion beam milling of an array of uniaxial grooves. They induce a washboard-like pinning potential landscape for vortices in the mixed state. By applying different magnetic fields, the most likely pinning sites along which the flux lines move through the samples have been selected. By this, either the background isotropic pinning of the pristine film or the enhanced isotropic pinning originating from the nanoprocessing has been probed. The enhanced pinning strength in the processed films has been found to correlate with the content of Ga implanted into the films during the nanopatterning.

Verification and implications of the multiple pin treatment in the SASSYS-1 LMR systems analysis code

International Nuclear Information System (INIS)

Dunn, F.E.

1994-01-01

As part of a program to obtain realistic, as opposed to excessively conservative, analysis of reactor transients, a multiple pin treatment for the analysis of intra-subassembly thermal hydraulics has been included in the SASSYS-1 liquid metal reactor systems analysis code. This new treatment has made possible a whole new level of verification for the code. The code can now predict the steady-state and transient responses of individual thermocouples within instrumented subassemlies in a reactor, rather than just predicting average temperatures for a subassembly. Very good agreement has been achieved between code predictions and the experimental measurements of steady-state and transient temperatures and flow rates in the Shutdown Heat Removal Tests in the EBR-II Reactor. Detailed multiple pin calculations for blanket subassemblies in the EBR-II reactor demonstrate that the actual steady-state and transient peak temperatures in these subassemblies are significantly lower than those that would be calculated by simpler models

PINS-3X Operations

Energy Technology Data Exchange (ETDEWEB)

E.H. Seabury

2013-09-01

Idaho National Laboratory’s (INL’s) Portable Isotopic Neutron Spectroscopy System (PINS) non-intrusively identifies the chemical fill of munitions and sealed containers. The PINS-3X variant of the system is used to identify explosives and uses a deuterium-tritium (DT) electronic neutron generator (ENG) as the neutron source. Use of the system, including possession and use of the neutron generator and shipment of the system components requires compliance with a number of regulations. This report outlines some of these requirements as well as some of the requirements in using the system outside of INL.

Elementary pinning force for a superconducting vortex

International Nuclear Information System (INIS)

Hyun, O.B.; Finnemore, D.K.; Schwartzkopf, L.; Clem, J.R.

1987-01-01

The elementary pinning force f/sub p/ has been measured for a single vortex trapped in one of the superconducting layers of a cross-strip Josephson junction. At temperatures close to the transition temperature the vortex can be pushed across the junction by a transport current. The vortex is found to move in a small number of discrete steps before it exits the junction. The pinning force for each site is found to be asymmetric and to have a value of about 10/sup -6/ N/m at the reduced temperature, t = T/T/sub c/ = 0.95. As a function of temperature, f/sub p/ is found to vary approximately as (1-t)/sup 3/2/. .AE

Cloning, expression and characterization of L-arabinose isomerase from Thermotoga neapolitana: bioconversion of D-galactose to D-tagatose using the enzyme.

Science.gov (United States)

Kim, Byoung-Chan; Lee, Yoon-Hee; Lee, Han-Seung; Lee, Dong-Woo; Choe, Eun-Ah; Pyun, Yu-Ryang

2002-06-18

Gene araA encoding an L-arabinose isomerase (AraA) from the hyperthermophile, Thermotoga neapolitana 5068 was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a polypeptide of 496 residues with a calculated molecular mass of 56677 Da. The deduced amino acid sequence has 94.8% identical amino acids compared with the residues in a putative L-arabinose isomerase of Thermotoga maritima. The recombinant enzyme expressed in E. coli was purified to homogeneity by heat treatment, ion exchange chromatography and gel filtration. The thermophilic enzyme had a maximum activity of L-arabinose isomerization and D-galactose isomerization at 85 degrees C, and required divalent cations such as Co(2+) and Mn(2+) for its activity and thermostability. The apparent K(m) values of the enzyme for L-arabinose and D-galactose were 116 mM (v(max), 119 micromol min(-1) mg(-1)) and 250 mM (v(max), 14.3 micromol min(-1) mg(-1)), respectively, that were determined in the presence of both 1 mM Co(2+) and 1 mM Mn(2+). A 68% conversion of D-galactose to D-tagatose was obtained using the recombinant enzyme at the isomerization temperature of 80 degrees C.

Application Of WIMS Code To Calculation Kartini Reactor Parameters By Pin-Cell And Cluster Method

International Nuclear Information System (INIS)

Sumarsono, Bambang; Tjiptono, T.W.

1996-01-01

Analysis UZrH fuel element parameters calculation in Kartini Reactor by WIMS Code has been done. The analysis is done by pin cell and cluster method. The pin cell method is done as a function percent burn-up and by 8 group 3 region analysis and cluster method by 8 group 12 region analysis. From analysis and calculation resulted K ∼ = 1.3687 by pin cell method and K ∼ = 1.3162 by cluster method and so deviation is 3.83%. By pin cell analysis as a function percent burn-up at the percent burn-up greater than 59.50%, the multiplication factor is less than one (k ∼ < 1) it is mean that the fuel element reactivity is negative

Dependence of J c on pinning center morphology: An explanation of record J c observed for discontinuous columnar pinning of vortices

International Nuclear Information System (INIS)

Weinstein, R.; Gandini, A.; Parks, D.; Sawh, R.; Mayes, B.

2006-01-01

An experimental study of defects caused by ion irradiation has proven that very discontinuous multiple-in-line-damage results in much higher J c than continuous columnar pinning centers. This is contrary to conventional wisdom. We develop a first approximation to a theory relating J c to the morphology of pinning centers. Qualitative agreement with experiment is achieved. Conclusions indicate the relative importance of pinning potential, percolation, and critical temperature for various morphologies. Results suggest the leading importance of pinning, compared to texture, weak links and oxygenation in achievement of the goal of high J c especially in large grain HTS

Nucleotide sequence of the triosephosphate isomerase gene from Macaca mulatta

Energy Technology Data Exchange (ETDEWEB)

Old, S.E.; Mohrenweiser, H.W. (Univ. of Michigan, Ann Arbor (USA))

1988-09-26

The triosephosphate isomerase gene from a rhesus monkey, Macaca mulatta, charon 34 library was sequenced. The human and chimpanzee enzymes differ from the rhesus enzyme at ASN 20 and GLU 198. The nucleotide sequence identity between rhesus and human is 97% in the coding region and >94% in the flanking regions. Comparison of the rhesus and chimp genes, including the intron and flanking sequences, does not suggest a mechanism for generating the two TPI peptides of proliferating cells from hominoids and a single peptide from the rhesus gene.

Development of a semiconductor neutron dosimeter with a PIN diode

International Nuclear Information System (INIS)

Kim, Seungho; Lee, Namho; Cho, Jaiwan; Youk, Geunuck

2004-01-01

When a Si PIN diode is exposed to fast neutrons, it produces displacement in Si lattice structure of the diode. Defects induced from structural dislocation become effective recombination centers for carriers which pass through the base of a PIN diode. Hence, increasing the resistivity of the diode decreases the current for the applied forward voltage. This paper involves the development of a neutron sensor based on the phenomena of the displacement effect damaged by neutron exposure. The neutron effect on the semiconductor was analyzed, and multi PIN diode arrays with various intrinsic layer (I layer) thicknesses and cross sections were fabricated. Under irradiation tests with a neutron beam, the manufactured diodes have good characteristics of linearity in a neutron irradiation experiment and give results that the increase of thickness of I layer and the decrease of the cross-section of the PIN diodes improve the sensitivity. Newly developed PIN diodes with a thicker I layer and various cross sections were retested and showed the best neutron sensitivity in the condition that the I layer thickness was similar to the length of a side of the cross-section. On the basis of two test results, final PIN diodes with a rectangular shape were manufactured and the characteristics for neutron detectors were analyzed through the neutron beam test using the on-line electronic dosimetry system. The developed PIN diode shows a good linearity to absorbed dose in the range of 0 to 1,000cGy (Tissue) and its neutron sensitivity is 13 mV/cGy at a constant current of 5 mA, that is three higher than that of similar commercially developed neutron detectors. Moreover the device shows less dependency on the orientation of the neutron beam and a considerable stability in an annealing test for a long period. (author)

Prevention of pin tract infection with titanium-copper alloys.

Science.gov (United States)

Shirai, Toshiharu; Tsuchiya, Hiroyuki; Shimizu, Tohru; Ohtani, Kaori; Zen, Yo; Tomita, Katsuro

2009-10-01

The most frequent complication in external fixation is pin tract infection. To reduce the incidence of implant-associated infection, many published reports have looked at preventing bacterial adhesion by treating the pin surface. This study aimed to evaluate the antibacterial activity of a Titanium-Copper (Ti-Cu) alloy on implant infection, and to determine the potential use of the Ti-Cu alloy as a biomaterial. Two forms of Ti-Cu alloys were synthesized: one with 1% Cu and the other with 5% Cu. For analyzing infectious behavior, the implants were exposed to Staphylococcus aureus and Escherichia coli. The reaction of pathogens to the Ti-Cu alloys was compared with their reaction to stainless steel and pure titanium as controls. Both Ti-Cu alloys evidently inhibited colonization by both bacteria. Conversely, cytocompatibility studies were performed using fibroblasts and colony formation on the metals was assessed by counting the number of colonies. Ti-1% Cu alloy showed no difference in the number of colonies compared with the control. External fixator pins made of Ti-Cu alloys were evaluated in a rabbit model. The tissue-implant interactions were analyzed for the presence of infection, inflammatory changes and osteoid-formation. Ti-1% Cu alloy significantly inhibited inflammation and infection, and had excellent osteoid-formation. Copper blood levels were measured before surgery and at 14 days postoperatively. Preoperative and postoperative blood copper values were not statistically different. Overall, it was concluded that Ti-Cu alloys have antimicrobial activity and substantially reduce the incidence of pin tract infection. Ti-1% Cu alloy shows particular promise as a biomaterial. (c) 2009 Wiley Periodicals, Inc.

l-Arabinose Isomerase and d-Xylose Isomerase from Lactobacillus reuteri: Characterization, Coexpression in the Food Grade Host Lactobacillus plantarum, and Application in the Conversion of d-Galactose and d-Glucose

Science.gov (United States)

2014-01-01

The l-arabinose isomerase (l-AI) and the d-xylose isomerase (d-XI) encoding genes from Lactobacillus reuteri (DSMZ 17509) were cloned and overexpressed in Escherichia coli BL21 (DE3). The proteins were purified to homogeneity by one-step affinity chromatography and characterized biochemically. l-AI displayed maximum activity at 65 °C and pH 6.0, whereas d-XI showed maximum activity at 65 °C and pH 5.0. Both enzymes require divalent metal ions. The genes were also ligated into the inducible lactobacillal expression vectors pSIP409 and pSIP609, the latter containing a food grade auxotrophy marker instead of an antibiotic resistance marker, and the l-AI- and d-XI-encoding sequences/genes were coexpressed in the food grade host Lactobacillus plantarum. The recombinant enzymes were tested for applications in carbohydrate conversion reactions of industrial relevance. The purified l-AI converted d-galactose to d-tagatose with a maximum conversion rate of 35%, and the d-XI isomerized d-glucose to d-fructose with a maximum conversion rate of 48% at 60 °C. PMID:24443973

Rational design of Bacillus stearothermophilus US100 L-arabinose isomerase: potential applications for D-tagatose production.

Science.gov (United States)

Rhimi, Moez; Aghajari, Nushin; Juy, Michel; Chouayekh, Hichem; Maguin, Emmanuelle; Haser, Richard; Bejar, Samir

2009-05-01

L-arabinose isomerases catalyze the bioconversion of D-galactose into D-tagatose. With the aim of producing an enzyme optimized for D-tagatose production, three Bacillus stearothermophilus US100 L-arabinose isomerase mutants were constructed, purified and characterized. Our results indicate that mutant Q268K was significantly more acidotolerant and more stable at acidic pH than the wild-type enzyme. The N175H mutant has a broad optimal temperature range from 50 to 65 degrees C. With the aim of constructing an acidotolerant mutant working at relatively low temperatures we generated the Q268K/N175H construct. This double mutant displays an optimal pH in the range 6.0-7.0 and an optimal activity around 50-65 degrees C, temperatures at which the enzyme was stable without addition of metal ions.

Fuel pin behavior under slow ramp-type transient-overpower conditions in the CABRI-FAST experiments

International Nuclear Information System (INIS)

Fukano, Yoshitaka; Onoda, Yuichi; Sato, Ikken; Charpenel, Jean

2009-01-01

In the CABRI-FAST experimental program, four in-pile tests were performed with slow power-ramp-type transient-overpower conditions (called hereafter as 'slow TOP') to study transient fuel pin behavior under inadvertent control rod withdrawal events in liquid metal cooled fast breeder reactors. Annular-pellet fuel pins were used in three tests, while a solid-pellet fuel pin was used in the other test. All of these pins were pre-irradiated in Phenix. The slow TOP test with a solid-pellet fuel pin was realized as a comparatory test against an existing test (E12) in the CABRI-2 program. In the CABRI-FAST test (BCF1), a power ramp rate of 3% Po/s was applied, while in the CABRI-2 test, 1% Po/s was adopted. Moreover, overpower condition was maintained for a few seconds beyond the observed pin failure in the BCF1 test. In spite of the different power ramp rates, evaluated fuel thermal conditions at the observed failure time are quite similar. The continued overpower condition in the BCF1 test resulted in gradual degradation of the pin structure providing information effective for evaluation of various accident scenarios. Three slow TOP tests with the annular fuel in the CABRI-FAST program resulted in no pin failure showing high failure threshold. Based on post-test examination data and a theoretical evaluation, it was concluded that intra-pin free spaces, such as central hole, macroscopic cracks and fuel-cladding gap effectively mitigated fuel cladding mechanical interaction. It was also clarified that cavity pressurization became effective only in case of very large amount of fuel melting. Furthermore, such cavity pressurization was effectively mitigated by a molten-fuel squirting into the upper blanket region pushing the blanket pellets upward. These CABRI FAST slow TOP tests, in combination with the existing CABRI and TREAT tests, provided an extended slow TOP test database with various fuel and transient conditions. (author)

Construction of phosphomannose isomerase (PMI) transformation vectors and evaluation of the effectiveness of vectors in tobacco (Nicotiana tabacum L).

Science.gov (United States)

Bahariah, Bohari; Parveez, Ghulam Kadir Ahmad; Masani, Mat Yunus Abdul; Khalid, Norzulaani

2012-01-01

Phosphomannose isomerase (pmi) gene isolated from Escherichia coli allows transgenic plants carrying it to convert mannose-6- phosphate (from mannose), a carbon source that could not be naturally utilized by plants into fructose-6-phosphate which can be utilized by plants as a carbon source. This conversion ability provides energy source to allow the transformed cells to survive on the medium containing mannose. In this study, four transformation vectors carrying the pmi gene alone or in combination with the β-glucuronidase (gusA) gene were constructed and driven by either the maize ubiquitin (Ubi1) or the cauliflower mosaic virus (CaMV35S) promoter. Restriction digestion, PCR amplification and sequencing were carried out to ensure sequence integrity and orientation. Tobacco was used as a model system to study the effectiveness of the constructs and selection system. PMI11G and pMI3G, which carry gusA gene, were used to study the gene transient expression in tobacco. PMI3 construct, which only carries the pmi gene driven by CaMV35S promoter, was stably transformed into tobacco using biolistics after selection on 30 g 1(-1) mannose without sucrose. Transgenic plants were verified using PCR analysis. PMI/pmi - Phosphomannose isomerase, Ubi1 - Maize ubiquitin promoter, CaMV35S - Cauliflower mosaic virus 35S promoter, gusA - β-glucuronidase GUS reporter gene.

Advanced control system for the Integral Fast Reactor fuel pin processor

International Nuclear Information System (INIS)

Lau, L.D.; Randall, P.F.; Benedict, R.W.; Levinskas, D.

1993-01-01

A computerized control system has been developed for the remotely-operated fuel pin processor used in the Integral Fast Reactor Program, Fuel Cycle Facility (FCF). The pin processor remotely shears cast EBR- reactor fuel pins to length, inspects them for diameter, straightness, length, and weight, and then inserts acceptable pins into new sodium-loaded stainless-steel fuel element jackets. Two main components comprise the control system: (1) a programmable logic controller (PLC), together with various input/output modules and associated relay ladder-logic associated computer software. The PLC system controls the remote operation of the machine as directed by the OCS, and also monitors the machine operation to make operational data available to the OCS. The OCS allows operator control of the machine, provides nearly real-time viewing of the operational data, allows on-line changes of machine operational parameters, and records the collected data for each acceptable pin on a central data archiving computer. The two main components of the control system provide the operator with various levels of control ranging from manual operation to completely automatic operation by means of a graphic touch screen interface

Clinical and histomorphometrical study on titanium dioxide-coated external fixation pins.

Science.gov (United States)

Koseki, Hironobu; Asahara, Tomohiko; Shida, Takayuki; Yoda, Itaru; Horiuchi, Hidehiko; Baba, Koumei; Osaki, Makoto

2013-01-01

Pin site infection is the most common and significant complication of external fixation. In this work, the efficacy of pins coated with titanium dioxide (TiO(2)) for inhibition of infection was compared with that of stainless steel control pins in an in vivo study. Pins contaminated with an identifiable Staphylococcus aureus strain were inserted into femoral bone in a rat model and exposed to ultraviolet A light for 30 minutes. On day 14, the animals were sacrificed and the bone and soft tissue around the pin were retrieved. The clinical findings and histological findings were evaluated in 60 samples. Clinical signs of infection were present in 76.7% of untreated pins, but in only 36.7% of TiO(2)-coated pins. The histological bone infection score and planimetric rate of occupation for bacterial colonies and neutrophils in the TiO(2)-coated pin group were lower than those in the control group. The bone-implant contact ratio of the TiO(2)-coated pin group was significantly higher (71.4%) than in the control pin group (58.2%). The TiO(2) was successful in decreasing infection both clinically and histomorphometrically. The photocatalytic bactericidal effect of TiO(2) is thought to be useful for inhibiting pin site infection after external fixation.

Fission rate distribution at the 84-pin radial section of a SVEA-96 Optima2 BWR assembly

Energy Technology Data Exchange (ETDEWEB)

Perret, Gregory; Murphy, Michael F.; Jatuff, Fabian [Paul Scherrer Institute, CH-5232 Villigen PSI (Switzerland); Chawla, Rakesh [Paul Scherrer Institute, CH-5232 Villigen PSI (Switzerland); Ecole Polytechnique Federale de Lausanne (EPFL), CH-1015 Lausanne (Switzerland)

2008-07-01

Westinghouse boiling water reactor SVEA-96 Optima2 assemblies were studied during the LWRPROTEUS program at the PROTEUS facility in the Paul Scherrer Institute. Measured radial fission rate distributions at the 84-pin elevation are compared with MCNPX predictions using both ENDF/B-VI (Release 2) and JEFF-3.1 data libraries. Predicted fission rates agree within +-4.5% using both libraries. Fission rates were over-predicted in UO{sub 2} pins close to the missing 1/3 pins and under-predicted in UO{sub 2} pins close to the missing 2/3 pins. Recurrent under-estimations were observed in the UO{sub 2}-Gd{sub 2}O{sub 3} pins, for both libraries, which might be explained by over-estimated thermal cross-sections of {sup 157}Gd, as suggested in a recent work of G. Leinweber et al. (2006). (authors)

Synchronizability on complex networks via pinning control

Indian Academy of Sciences (India)

Keywords. Complex network; the pinning synchronization; synchronizability. ... The findings reveal the relationship between the decreasing speed of maximum eigenvalue sequence of the principal submatrices for coupling matrix and the synchronizability on complex networks via pinning control. We discuss the ...

The "Good, Bad and Ugly" pin site grading system: A reliable and memorable method for documenting and monitoring ring fixator pin sites.

Science.gov (United States)

Clint, S A; Eastwood, D M; Chasseaud, M; Calder, P R; Marsh, D R

2010-02-01

Although there is much in the literature regarding pin site infections, there is no accepted, validated method for documenting their state. We present a system for reliably labelling pin sites on any ring fixator construct and an easy-to-remember grading system to document the state of each pin site. Each site is graded in terms of erythema, pain and discharge to give a 3-point scale, named "Good", "Bad" and "Ugly" for ease of recall. This system was tested for intra- and inter-observer reproducibility. 15 patients undergoing elective limb reconstruction were recruited. A total of 218 pin sites were independently scored by 2 examiners. 82 were then re-examined later by the same examiners. 514 pin sites were felt to be "Good", 80 "Bad" and 6 "Ugly". The reproducibility of the system was found to be excellent. We feel our system gives a quick, reliable and reproducible method to monitor individual pin sites and their response to treatment. Crown Copyright 2009. Published by Elsevier Ltd. All rights reserved.

Spectroscopic amplifier for pin diode; Amplificador espectroscopico para diodo Pin

Energy Technology Data Exchange (ETDEWEB)

Alonso M, M. S.; Hernandez D, V. M.; Vega C, H. R., E-mail: bebe.luna_s@hotmail.com [Universidad Autonoma de Zacatecas, Unidad Academica de Estudios Nucleares, Cipres No. 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)

2014-10-15

The photodiode remains the basic choice for the photo-detection and is widely used in optical communications, medical diagnostics and field of corpuscular radiation. In detecting radiation it has been used for monitoring radon and its progeny and inexpensive spectrometric systems. The development of a spectroscopic amplifier for Pin diode is presented which has the following characteristics: canceler Pole-Zero (P/Z) with a time constant of 8 μs; constant gain of 57, suitable for the acquisition system; 4th integrator Gaussian order to waveform change of exponential input to semi-Gaussian output and finally a stage of baseline restorer which prevents Dc signal contribution to the next stage. The operational amplifier used is the TLE2074 of BiFET technology of Texas Instruments with 10 MHz bandwidth, 25 V/μs of slew rate and a noise floor of 17 nv/(Hz)1/2. The integrated circuit has 4 operational amplifiers and in is contained the total of spectroscopic amplifier that is the goal of electronic design. The results show like the exponential input signal is converted to semi-Gaussian, modifying only the amplitude according to the specifications in the design. The total system is formed by the detector, which is the Pin diode, a sensitive preamplifier to the load, the spectroscopic amplifier that is what is presented and finally a pulse height analyzer (Mca) which is where the spectrum is shown. (Author)

A pin-assisted retention technique for resin-bonded restorations.

Science.gov (United States)

Miara, P; Touati, B

1992-09-01

The value of pins for auxiliary retention has been demonstrated many times. The use of pins with resin-bonded restorations allows for improved aesthetics and less tooth reduction while increasing resistance to dislodging forces. Clinical and technical procedures for resin-bonded bridges with pin-assisted retention are presented.

Ethanol production from lignocellulosic hydrolysates using engineered Saccharomyces cerevisiae harboring xylose isomerase-based pathway.

Science.gov (United States)

Ko, Ja Kyong; Um, Youngsoon; Woo, Han Min; Kim, Kyoung Heon; Lee, Sun-Mi

2016-06-01

The efficient co-fermentation of glucose and xylose is necessary for the economically feasible bioethanol production from lignocellulosic biomass. Even with xylose utilizing Saccharomyces cerevisiae, the efficiency of the lignocellulosic ethanol production remains suboptimal mainly due to the low conversion yield of xylose to ethanol. In this study, we evaluated the co-fermentation performances of SXA-R2P-E, a recently engineered isomerase-based xylose utilizing strain, in mixed sugars and in lignocellulosic hydrolysates. In a high-sugar fermentation with 70g/L of glucose and 40g/L of xylose, SXA-R2P-E produced 50g/L of ethanol with an yield of 0.43gethanol/gsugars at 72h. From dilute acid-pretreated hydrolysates of rice straw and hardwood (oak), the strain produced 18-21g/L of ethanol with among the highest yield of 0.43-0.46gethanol/gsugars ever reported. This study shows a highly promising potential of a xylose isomerase-expressing strain as an industrially relevant ethanol producer from lignocellulosic hydrolysates. Copyright © 2016 Elsevier Ltd. All rights reserved.

Method and apparatus for enhancing vortex pinning by conformal crystal arrays

Science.gov (United States)

Janko, Boldizsar; Reichhardt, Cynthia; Reichhardt, Charles; Ray, Dipanjan

2015-07-14

Disclosed is a method and apparatus for strongly enhancing vortex pinning by conformal crystal arrays. The conformal crystal array is constructed by a conformal transformation of a hexagonal lattice, producing a non-uniform structure with a gradient where the local six-fold coordination of the pinning sites is preserved, and with an arching effect. The conformal pinning arrays produce significantly enhanced vortex pinning over a much wider range of field than that found for other vortex pinning geometries with an equivalent number of vortex pinning sites, such as random, square, and triangular.

The treatment of burnable poison pins in LWRWIMS

International Nuclear Information System (INIS)

Halsall, M.J.

1982-12-01

This report describes an investigation into the modelling approximations normally made when the LWR lattice code LWRWIMS is used for design calculations on assemblies containing burnable poison pins. Parameters investigated include energy group structure, intervals between calculations in MWd/te and spatial subdivision of the poison pins. An estimate is made of the effect of using pin-cell smearing with diffusion theory for the assembly geometry, instead of a more exact heterogeneous transport theory calculation. The influence on reactivity of the minor gadolinium isotopes 152, 154, 156, 158 and 160 in a poison pin dominated by the isotopes 155 and 157 is presented, and finally, recommendations on the use of LWRWIMS for this type of calculation are made. (author)

New Universal Tribometer as Pin or Ball-on-Disc and Reciprocating Pin-on-Plate Types

Directory of Open Access Journals (Sweden)

H. Kaleli

2016-06-01

Full Text Available The present paper contains a description of a new Universal Tribometer design which enables simulation of different contact and test types such as pin-on-disc, ball-on-disc and linear reciprocating tests. There are many models of wear Tribometer in the world market. These devices are manufactured by various companies abroad and are imported to our country. Cost of this devices start from 50.000 euros and goes to hundreds of thousands of euros. One of the most commonly used of this device is Reciprocating Pin-on-Plate Tribo Test Machine. This wear tester is produced at a low limited cost within the KAP (Scientifical Research Project Coordinator of Yıldız Technical University. The test machine can work including three types of Tribotest rigs (Reciprocating Pin-on-Plate, Pin-on-Disc and Ball-on-Disc. It is designed to operate also at high temperatures up to 500 ˚C. The new piece of equipment allows instrumented tribological testing of piston ring and cylinder liner samples at low and high temperatures and boundary lubrication conditions of any typical gasoline or Diesel engines. Some friction results were shown in boundary lubricating conditions between piston ring and cylinder liner sliding pairs describing Tribotest machine is driven by AC servo motor which is more accurate than DC motor.

Primary hip spica with crossed retrograde intramedullary rush pins ...

African Journals Online (AJOL)

Bursitis and penetration of pins at the site of Rush pin insertion is a complication associated with this method of treatment. Conclusion: Closed reduction and internal fixation with crossed Rush pins was a superior treatment method in terms of early weight bearing and restoration of normal anatomy. Keywords: Femoral ...

Power release estimation inside of a fuel pin neighbouring a WWER-440 control rod

International Nuclear Information System (INIS)

Mikus, J.

2006-01-01

This work presents an estimation of the control rod (CR) influence in the WWER-440 core on the power release inside of a fuel pin neighbouring CR, that can have some consequences due to possible static and cyclic loads, for example fuel pin / fuel assembly bowing. For this purpose detailed (usual) axial power distribution measurements were performed in a WWER-440 type core on the light water, zero-power research reactor LR-0 in fuel pins near to an authentic CR model at zero boron concentration in moderator, modelling the conditions at the end of fuel cycle. To demonstrate the CR influence on power distribution inside of one fuel pin neighbouring CR, results of above measurements were used for estimation of the: 1) Axial power distribution inside of the investigated fuel pin in both opposite positions on its pellets surface that are situated to- and outwards CR and corresponding gradient of the (r, z) - power distribution in above opposite positions and 2) Azimuthal power distributions on pellet surface of the investigated fuel pin in horizontal planes at selected axial coordinates. Similar information can be relevant from the viewpoint of the fuel pin failures occurrence investigation

A three-dimensional pin-wise analysis for CEA ejection accident

Energy Technology Data Exchange (ETDEWEB)

Park, Guen-Tae; Park, Min-Ho; Park, Jin-Woo; Um, Kil-Sup; Choi, Tong-Soo [KEPCO NF, Daejeon (Korea, Republic of)

2016-10-15

The ejection of a control element assembly (CEA) with high reactivity worth causes the sudden insertion of reactivity into the core. Immediately after the CEA ejection, the nuclear power of the reactor dramatically increases in an exponential behavior until the doppler effect becomes important and turns the reactivity balance and power down to lower levels. The 3-D CEA ejection analysis methodology has been developed using the multi-dimensional code coupling system, CHASER, which couples three dimensional core neutron kinetics code ASTRA, subchannel analysis code THALES, and fuel performance analysis code FROST using message passing interface (MPI). This paper presents the pin-by-pin level analysis result with the 3-D CEA ejection analysis methodology using the CHASER. The pin-by-pin level analysis consists of DNBR, enthalpy and Pellet/Clad Mechanical Interaction (PCMI) analysis. All the evaluations are simulated for APR1400 plant loaded with PLUS7 fuel. In this paper, the pin-by-pin analysis using the multidimensional core transient code, CHASER, is presented with respect to enthalpy, DNBR and PCMI for APR1400 plant loaded with PLUS7 fuel. For the pin-by-pin enthalpy and DNBR analysis, the quarter core for HFP case or 15 - 20 assemblies around the most severe assembly for part powers or HZP cases are selected. And PCMI calculation is performed for all the rods in the whole core during a conservative time period. The pin-by-pin analysis results show that the regulatory guidelines of CEA ejection accident are satisfied.

Interaction of PLS and PIN and hormonal crosstalk in Arabidopsis root developmentHormonal crosstalk in Arabidopsis

Directory of Open Access Journals (Sweden)

Junli eLiu

2013-04-01

Full Text Available Understanding how hormones and genes interact to coordinate plant growth is a major challenge in developmental biology. The activities of auxin, ethylene and cytokinin depend on cellular context and exhibit either synergistic or antagonistic interactions. Here we use experimentation and network construction to elucidate the role of the interaction of the POLARIS peptide (PLS and the auxin efflux carrier PIN proteins in the crosstalk of three hormones (auxin, ethylene and cytokinin in Arabidopsis root development. In ethylene hypersignalling mutants such as polaris (pls, we show experimentally that expression of both PIN1 and PIN2 significantly increases. This relationship is analysed in the context of the crosstalk between auxin, ethylene and cytokinin: in pls, endogenous auxin, ethylene and cytokinin concentration decreases, approximately remains unchanged and increases, respectively. Experimental data are integrated into a hormonal crosstalk network through combination with information in literature. Network construction reveals that the regulation of both PIN1 and PIN2 is predominantly via ethylene signalling. In addition, it is deduced that the relationship between cytokinin and PIN1 and PIN2 levels implies a regulatory role of cytokinin in addition to its regulation to auxin, ethylene and PLS levels. We discuss how the network of hormones and genes coordinates plant growth by simultaneously regulating the activities of auxin, ethylene and cytokinin signalling pathways.