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Sample records for chitinase insertion domain

  1. Biochemistry of plant class IV chitinases and fungal chitinase-modifying proteins

    Science.gov (United States)

    Plant class IV chitinases have 2 domains, a small (3 kDa) amino-terminal domain with homology to carbohydrate binding peptides, and a larger (25 kDa) catalytic domain. The biological function of these chitinases is not known. But it is known that some pathogenic fungi secrete chitinase modifying pro...

  2. Lesion insertion in the projection domain: Methods and initial results

    International Nuclear Information System (INIS)

    Purpose: To perform task-based image quality assessment in CT, it is desirable to have a large number of realistic patient images with known diagnostic truth. One effective way of achieving this objective is to create hybrid images that combine patient images with inserted lesions. Because conventional hybrid images generated in the image domain fails to reflect the impact of scan and reconstruction parameters on lesion appearance, this study explored a projection-domain approach. Methods: Lesions were segmented from patient images and forward projected to acquire lesion projections. The forward-projection geometry was designed according to a commercial CT scanner and accommodated both axial and helical modes with various focal spot movement patterns. The energy employed by the commercial CT scanner for beam hardening correction was measured and used for the forward projection. The lesion projections were inserted into patient projections decoded from commercial CT projection data. The combined projections were formatted to match those of commercial CT raw data, loaded onto a commercial CT scanner, and reconstructed to create the hybrid images. Two validations were performed. First, to validate the accuracy of the forward-projection geometry, images were reconstructed from the forward projections of a virtual ACR phantom and compared to physically acquired ACR phantom images in terms of CT number accuracy and high-contrast resolution. Second, to validate the realism of the lesion in hybrid images, liver lesions were segmented from patient images and inserted back into the same patients, each at a new location specified by a radiologist. The inserted lesions were compared to the original lesions and visually assessed for realism by two experienced radiologists in a blinded fashion. Results: For the validation of the forward-projection geometry, the images reconstructed from the forward projections of the virtual ACR phantom were consistent with the images physically

  3. Lesion insertion in the projection domain: Methods and initial results

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Baiyu; Leng, Shuai; Yu, Lifeng; Yu, Zhicong; Ma, Chi; McCollough, Cynthia, E-mail: mccollough.cynthia@mayo.edu [Department of Radiology, Mayo Clinic, Rochester, Minnesota 55905 (United States)

    2015-12-15

    Purpose: To perform task-based image quality assessment in CT, it is desirable to have a large number of realistic patient images with known diagnostic truth. One effective way of achieving this objective is to create hybrid images that combine patient images with inserted lesions. Because conventional hybrid images generated in the image domain fails to reflect the impact of scan and reconstruction parameters on lesion appearance, this study explored a projection-domain approach. Methods: Lesions were segmented from patient images and forward projected to acquire lesion projections. The forward-projection geometry was designed according to a commercial CT scanner and accommodated both axial and helical modes with various focal spot movement patterns. The energy employed by the commercial CT scanner for beam hardening correction was measured and used for the forward projection. The lesion projections were inserted into patient projections decoded from commercial CT projection data. The combined projections were formatted to match those of commercial CT raw data, loaded onto a commercial CT scanner, and reconstructed to create the hybrid images. Two validations were performed. First, to validate the accuracy of the forward-projection geometry, images were reconstructed from the forward projections of a virtual ACR phantom and compared to physically acquired ACR phantom images in terms of CT number accuracy and high-contrast resolution. Second, to validate the realism of the lesion in hybrid images, liver lesions were segmented from patient images and inserted back into the same patients, each at a new location specified by a radiologist. The inserted lesions were compared to the original lesions and visually assessed for realism by two experienced radiologists in a blinded fashion. Results: For the validation of the forward-projection geometry, the images reconstructed from the forward projections of the virtual ACR phantom were consistent with the images physically

  4. Insertion of tetracysteine motifs into dopamine transporter extracellular domains.

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    Deanna M Navaroli

    Full Text Available The neuronal dopamine transporter (DAT is a major determinant of extracellular dopamine (DA levels and is the primary target for a variety of addictive and therapeutic psychoactive drugs. DAT is acutely regulated by protein kinase C (PKC activation and amphetamine exposure, both of which modulate DAT surface expression by endocytic trafficking. In order to use live imaging approaches to study DAT endocytosis, methods are needed to exclusively label the DAT surface pool. The use of membrane impermeant, sulfonated biarsenic dyes holds potential as one such approach, and requires introduction of an extracellular tetracysteine motif (tetraCys; CCPGCC to facilitate dye binding. In the current study, we took advantage of intrinsic proline-glycine (Pro-Gly dipeptides encoded in predicted DAT extracellular domains to introduce tetraCys motifs into DAT extracellular loops 2, 3, and 4. [(3H]DA uptake studies, surface biotinylation and fluorescence microscopy in PC12 cells indicate that tetraCys insertion into the DAT second extracellular loop results in a functional transporter that maintains PKC-mediated downregulation. Introduction of tetraCys into extracellular loops 3 and 4 yielded DATs with severely compromised function that failed to mature and traffic to the cell surface. This is the first demonstration of successful introduction of a tetracysteine motif into a DAT extracellular domain, and may hold promise for use of biarsenic dyes in live DAT imaging studies.

  5. Latex-allergic patients sensitized to the major allergen hevein and hevein-like domains of class I chitinases show no increased frequency of latex-associated plant food allergy

    OpenAIRE

    Radauer, Christian; Adhami, Farzaneh; Fürtler, Irene; Wagner, Stefan; Allwardt, Dorothee; Scala, Enrico; Ebner, Christof; Hafner, Christine; Hemmer, Wolfgang; Mari, Adriano; Breiteneder, Heimo

    2011-01-01

    Allergies to certain fruits such as banana, avocado, chestnut and kiwi are described in 30–70% of latex-allergic patients. This association is attributed to the cross-reactivity between the major latex allergen hevein and hevein-like domains (HLDs) from fruit class I chitinases. We aimed to assess the extent of cross-reactivity between hevein and HLDs using sera from latex-allergic patients with and without plant food allergy. Hevein and HLDs of latex, banana, and avocado chitinases were expr...

  6. Crystallization and preliminary X-ray diffraction studies of the catalytic domain of a novel chitinase, a member of GH family 23, from the moderately thermophilic bacterium Ralstonia sp. A-471

    International Nuclear Information System (INIS)

    The catalytic domain of a novel chitinase, which is a member of GH family 23, from the moderately thermophilic bacterium Ralstonia sp. A-471 was crystallized and diffraction data were collected to 1.85 Å resolution. Chitinase from the moderately thermophilic bacterium Ralstonia sp. A-471 (Ra-ChiC) is divided into two domains: a chitin-binding domain (residues 36–80) and a catalytic domain (residues 103–252). Although the catalytic domain of Ra-ChiC has homology to goose-type lysozyme, Ra-ChiC does not show lysozyme activity but does show chitinase activity. The catalytic domain with part of an interdomain loop (Ra-ChiC89–252) was crystallized under several different conditions using polyethylene glycol as a precipitant. The crystals diffracted to 1.85 Å resolution and belonged to space group P6122 or P6522, with unit-cell parameters a = b = 100, c = 243 Å. The calculated Matthews coefficient was approximately 3.2, 2.4 or 1.9 Å3 Da−1 assuming the presence of three, four or five Ra-ChiC89–252 molecules in the asymmetric unit, respectively

  7. Chitinases: An update

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    Rifat Hamid

    2013-01-01

    Full Text Available Chitin, the second most abundant polysaccharide in nature after cellulose, is found in the exoskeleton of insects, fungi, yeast, and algae, and in the internal structures of other vertebrates. Chitinases are enzymes that degrade chitin. Chitinases contribute to the generation of carbon and nitrogen in the ecosystem. Chitin and chitinolytic enzymes are gaining importance for their biotechnological applications, especially the chitinases exploited in agriculture fields to control pathogens. Chitinases have a use in human health care, especially in human diseases like asthma. Chitinases have wide-ranging applications including the preparation of pharmaceutically important chitooligosaccharides and N-acetyl D glucosamine, preparation of single-cell protein, isolation of protoplasts from fungi and yeast, control of pathogenic fungi, treatment of chitinous waste, mosquito control and morphogenesis, etc. In this review, the various types of chitinases and the chitinases found in different organisms such as bacteria, plants, fungi, and mammals are discussed.

  8. Chitinases: An update

    OpenAIRE

    Rifat Hamid; Minhaj A Khan; Mahboob Ahmad; Malik Mobeen Ahmad; Malik Zainul Abdin; Javed Musarrat; Saleem Javed

    2013-01-01

    Chitin, the second most abundant polysaccharide in nature after cellulose, is found in the exoskeleton of insects, fungi, yeast, and algae, and in the internal structures of other vertebrates. Chitinases are enzymes that degrade chitin. Chitinases contribute to the generation of carbon and nitrogen in the ecosystem. Chitin and chitinolytic enzymes are gaining importance for their biotechnological applications, especially the chitinases exploited in agriculture fields to control pathogens. Chi...

  9. Evaluation of a projection-domain lung nodule insertion technique in thoracic CT

    Science.gov (United States)

    Ma, Chi; Chen, Baiyu; Koo, Chi Wan; Takahashi, Edwin A.; Fletcher, Joel G.; McCollough, Cynthia H.; Levin, David L.; Kuzo, Ronald S.; Viers, Lyndsay D.; Vincent Sheldon, Stephanie A.; Leng, Shuai; Yu, Lifeng

    2016-04-01

    Task-based assessment of computed tomography (CT) image quality requires a large number of cases with ground truth. Inserting lesions into existing cases to simulate positive cases is a promising alternative approach. The aim of this study was to evaluate a recently-developed raw-data based lesion insertion technique in thoracic CT. Lung lesions were segmented from patient CT images, forward projected, and reinserted into the same patient CT projection data. In total, 32 nodules of various attenuations were segmented from 21 CT cases. Two experienced radiologists and 2 residents blinded to the process independently evaluated these inserted nodules in two sub-studies. First, the 32 inserted and the 32 original nodules were presented in a randomized order and each received a rating score from 1 to 10 (1=absolutely artificial to 10=absolutely realistic). Second, the inserted and the corresponding original lesions were presented side-by-side to each reader, who identified the inserted lesion and provided a confidence score (1=no confidence to 5=completely certain). For the randomized evaluation, discrimination of real versus artificial nodules was poor with areas under the receiver operative characteristic curves being 0.69 (95% CI: 0.58-0.78), 0.57 (95% CI: 0.46-0.68), and 0.62 (95% CI: 0.54-0.69) for the 2 radiologists, 2 residents, and all 4 readers, respectively. For the side-by-side evaluation, although all 4 readers correctly identified inserted lesions in 103/128 pairs, the confidence score was moderate (2.6). Our projection-domain based lung nodule insertion technique provides a robust method to artificially generate clinical cases that prove to be difficult to differentiate from real cases.

  10. Chitinase from Serratia marcescens

    International Nuclear Information System (INIS)

    This paper discusses the chitinase which is assayed by the liberation of tritiated oligosaccharides from [acetyl-3H]chitin,3 with phosphate buffer at pH 6.3, at a final concentration of 0.05 M in the reaction mixture (buffer A). The author explains that a unit of chitinase is that amount of enzyme which catalyzes the release of 1 μmol of soluble product (calculated as N- acetylglucosamine) in 1 min at 30 degrees

  11. Cloning, Expression and 3D Structure Prediction of Chitinase from Chitinolyticbacter meiyuanensis SYBC-H1

    Science.gov (United States)

    Hao, Zhikui; Wu, Hangui; Yang, Meiling; Chen, Jianjun; Xi, Limin; Zhao, Weijie; Yu, Jialin; Liu, Jiayang; Liao, Xiangru; Huang, Qingguo

    2016-01-01

    Two CHI genes from Chitinolyticbacter meiyuanensis SYBC-H1 encoding chitinases were identified and their protein 3D structures were predicted. According to the amino acid sequence alignment, CHI1 gene encoding 166 aa had a structural domain similar to the GH18 type II chitinase, and CHI2 gene encoding 383 aa had the same catalytic domain as the glycoside hydrolase family 19 chitinase. In this study, CHI2 chitinase were expressed in Escherichia coli BL21 cells, and this protein was purified by ammonium sulfate precipitation, DEAE-cellulose, and Sephadex G-100 chromatography. Optimal activity of CHI2 chitinase occurred at a temperature of 40 °C and a pH of 6.5. The presence of metal ions Fe3+, Fe2+, and Zn2+ inhibited CHI2 chitinase activity, while Na+ and K+ promoted its activity. Furthermore, the presence of EGTA, EDTA, and β-mercaptoethanol significantly increased the stability of CHI2 chitinase. The CHI2 chitinase was active with p-NP-GlcNAc, with the Km and Vm values of 23.0 µmol/L and 9.1 mM/min at a temperature of 37 °C, respectively. Additionally, the CHI2 chitinase was characterized as an N-acetyl glucosaminidase based on the hydrolysate from chitin. Overall, our results demonstrated CHI2 chitinase with remarkable biochemical properties is suitable for bioconversion of chitin waste. PMID:27240345

  12. STRUCTURAL FEATURES OF PLANT CHITINASES AND CHITIN-BINDING PROTEINS

    NARCIS (Netherlands)

    BEINTEMA, JJ

    1994-01-01

    Structural features of plant chitinases and chitin-binding proteins are discussed. Many of these proteins consist of multiple domains,of which the chitin-binding hevein domain is a predominant one. X-ray and NMR structures of representatives of the major classes of these proteins are available now,

  13. Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    OpenAIRE

    Davidson, John F.; Fox, Richard; Harris, Dawn D.; Lyons-Abbott, Sally; Loeb, Lawrence A.

    2003-01-01

    Insertion of the T3 DNA polymerase thioredoxin binding domain (TBD) into the distantly related thermostable Taq DNA polymerase at an analogous position in the thumb domain, converts the Taq DNA polymerase from a low processive to a highly processive enzyme. Processivity is dependent on the presence of thioredoxin. The enhancement in processivity is 20–50-fold when compared with the wild-type Taq DNA polymerase or to the recombinant polymerase in the absence of thioredoxin. The recombinant Taq...

  14. pH-Triggered Conformational Switching along the Membrane Insertion Pathway of the Diphtheria Toxin T-Domain

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    Alexey S. Ladokhin

    2013-08-01

    Full Text Available The translocation (T-domain plays a key role in the action of diphtheria toxin and is responsible for transferring the catalytic domain across the endosomal membrane into the cytosol in response to acidification. Deciphering the molecular mechanism of pH-dependent refolding and membrane insertion of the T-domain, which is considered to be a paradigm for cell entry of other bacterial toxins, reveals general physicochemical principles underlying membrane protein assembly and signaling on membrane interfaces. Structure-function studies along the T-domain insertion pathway have been affected by the presence of multiple conformations at the same time, which hinders the application of high-resolution structural techniques. Here, we review recent progress in structural, functional and thermodynamic studies of the T-domain archived using a combination of site-selective fluorescence labeling with an array of spectroscopic techniques and computer simulations. We also discuss the principles of conformational switching along the insertion pathway revealed by studies of a series of T-domain mutants with substitutions of histidine residues.

  15. Cloning of the Bacillus thuringiensis serovar sotto chitinase (Schi gene and characterization of its protein

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    Wan-Fang Zhong

    2005-12-01

    Full Text Available Chitinase plays a positive role in the pathogenicity of Bacillus thuringiensis to insect pests. We used touchdown PCR to clone the chitinase (Schi gene from Bacillus thuringiensis serovar sotto (Bt sotto chromosomal DNA. Our DNA sequencing analysis revealed that the Bt sotto Schi gene consists of an open reading frame (ORF of 2067 nucleotides with codes for the chitinase precursor. We also found that the putative promoter consensus sequences (the -35 and -10 regions of the Bt soto Schi gene are identical to those of the chiA71 gene from Bt Pakistani, the chiA74 gene from Bt kenyae and the ichi gene from Bt israelensis. The Schi chitinase precursor is 688 amino acids long with an estimated molecular mass of 75.75 kDa and a theoretical isoelectric point of 5.74, and contains four domains, which are, in sequence, a signal peptide, an N-terminal catalytic domain, a fibronectin type III like domain and a C-terminal chitin-binding domain. Sequence comparison and the evolutionary relationship of the Bt sotto Schi chitinase to other chitinase and chitinase-like proteins are also discussed.

  16. Characterization of a chitinase from the cellulolytic actinomycete Thermobifida fusca.

    Science.gov (United States)

    Gaber, Yasser; Mekasha, Sophanit; Vaaje-Kolstad, Gustav; Eijsink, Vincent G H; Fraaije, Marco W

    2016-09-01

    Thermobifida fusca is a well-known cellulose-degrading actinomycete, which produces various glycoside hydrolases for this purpose. However, despite the presence of putative chitinase genes in its genome, T. fusca has not been reported to grow on chitin as sole carbon source. In this study, a gene encoding a putative membrane-anchored GH18 chitinase (Tfu0868) from T. fusca has been cloned and overexpressed in Escherichia coli. The protein was produced as SUMO fusion protein and, upon removal of the SUMO domain, soluble pure TfChi18A was obtained with yields typically amounting to 150mg per litre of culture. The enzyme was found to be relatively thermostable (apparent Tm=57.5°C) but not particularly thermoactive, the optimum temperature being 40-45°C. TfChi18A bound to α- and β-chitin and degraded both these substrates. Interestingly, activity towards colloidal chitin was minimal and in this case, substrate inhibition was observed. TfChi18A also cleaved soluble chito-oligosaccharides and showed a clear preference for substrates having five sugars or more. While these results show that TfChi18A is a catalytically competent GH18 chitinase, the observed catalytic rates were low compared to those of well-studied GH18 chitinases. This suggests that TfChi18A is not a true chitinase and not likely to endow T. fusca with the ability to grow on chitin. PMID:27108953

  17. Stabilizing the integrin αM inserted domain in alternative conformations with a range of engineered disulfide bonds

    OpenAIRE

    Shimaoka, Motomu; Lu, Chafen; Salas, Azucena; Xiao, Tsan; Takagi, Junichi; Springer, Timothy A.

    2002-01-01

    Conformational movement of the C-terminal α7 helix in the integrin inserted (I) domain, a major ligand-binding domain that adopts an α/β Rossmann fold, has been proposed to allosterically regulate ligand-binding activity. Disulfide bonds were engineered here to reversibly lock the position of the α7 helix in one of two alternative conformations seen in crystal structures, termed open and closed. Our results show that pairs of residues with Cβ atoms farther apart than optimal for disulfide bon...

  18. Genetic Variations of Kinase Inserts Domain Receptor (KDR) Gene Are Associated with the Risk of Astrocytomas.

    Science.gov (United States)

    Gao, Yufei; Ma, Piyong; He, Yichun; Liu, Yan; Jiang, Yang

    2016-05-01

    Astrocytomas is one of the most common central nervous system (CNS) tumors with high mortality rate. Kinase insert domain receptor (KDR) is involved in the regulation of tumor angiogenesis, migration, and vascular permeability. The aim of the study was to explore the relationship between KDR polymorphisms and risk of astrocytomas. Blood samples were collected from 157 astrocytomas patients and 160 healthy controls. Three tag-SNPs (rs2071559C/T, rs2305948T/C, and rs1870377A/T) were identified from the International HapMap Project Databases and genotyped using the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). We evaluated the astrocytomas risk caused by individual SNPs and haplotype using odds ratios (ORs) and their 95 % confidence intervals (CIs). In the overall individual SNP analysis, the C allele of rs2071559 was correlated with an increased risk of astrocytomas. However, individuals with mutant allele A and genotype TA + AA of rs1870377 showed a protective effect against astrocytomas. Subgroup analysis based on WHO tumor grade revealed that the C allele of rs2071559 had more influence with the risk of astrocytomas in the grade III-IV (OR = 1.91) subgroup than the grade I-II (OR = 1.47) group. Genotype TT of rs2305948 was found to be significantly associated with susceptibility of astrocytomas only in the grade III-IV subgroup. The protective effect of rs1870377 did not reveal significant difference between the grade III-IV and grade I-II subgroups. Meanwhile, stratified analysis demonstrated that mutation of rs2071559 and rs2305948 could elevate the risk of astrocytomas more significantly in the subgroup of smokers than the nonsmokers. Interestingly, the protective effect of rs1870377 was more obvious in the nonsmokers than the smokers. Additionally, haplotype-specific analysis showed that haplotype CCT and CTT were related with an increased risk of astrocytomas. We found that individual with variants of rs

  19. Higher-Resolution Structure of the Human Insulin Receptor Ectodomain: Multi-Modal Inclusion of the Insert Domain.

    Science.gov (United States)

    Croll, Tristan I; Smith, Brian J; Margetts, Mai B; Whittaker, Jonathan; Weiss, Michael A; Ward, Colin W; Lawrence, Michael C

    2016-03-01

    Insulin receptor (IR) signaling is critical to controlling nutrient uptake and metabolism. However, only a low-resolution (3.8 Å) structure currently exists for the IR ectodomain, with some segments ill-defined or unmodeled due to disorder. Here, we revise this structure using new diffraction data to 3.3 Å resolution that allow improved modeling of the N-linked glycans, the first and third fibronectin type III domains, and the insert domain. A novel haptic interactive molecular dynamics strategy was used to aid fitting to low-resolution electron density maps. The resulting model provides a foundation for investigation of structural transitions in IR upon ligand binding. PMID:26853939

  20. High-level expression and characterization of two chitinases, ChiCH and ChiCW, of Bacillus cereus 28-9 in Escherichia coli

    International Nuclear Information System (INIS)

    Many chitinase genes have been cloned and sequenced from prokaryotes and eukaryotes but overexpression of chitinases in Escherichia coli cells was less reported. ChiCH and ChiCW of Bacillus cereus 28-9 belong to two distinct groups based on their amino acid sequences of catalytic domains, and in addition, domain structures of two enzymes are different. In this study, we established an ideal method for high-level expression of chitinases in E. coli as glutathione-S-transferase fusion proteins using pGEX-6P-1 vector. Both ChiCH and ChiCW were successfully highly expressed in E. coli cells as soluble GST-chitinase fusion proteins, and recombinant native ChiCH and ChiCW could be purified after cleavage with PreScission protease to remove GST tag. Purified chitinases were used for biochemical characterization of kinetics, hydrolysis products, and binding activities. The results indicate that ChiCW is an endo-chitinase and effectively hydrolyzes chitin and chito-multimers to chito-oligomers and the end product chitobiose, and ChiCH is an exo-chitinase and degrades chito-oligomers to produce chitobiose. Furthermore, due to higher affinity of ChiCW toward colloidal chitin than Avicel, C-terminal domain of ChiCW should be classified as a chitin-binding domain not a cellulose-binding domain although that was revealed as a cellulose-binding domain by conserved domain analysis. Therefore, the method of high-level expression of chitinases is helpful to studies and applications of chitinases

  1. NSCLC harboring EGFR exon-20 insertions after the regulatory C-helix of kinase domain responds poorly to known EGFR inhibitors.

    Science.gov (United States)

    Yang, Mengmeng; Xu, Xiaoxi; Cai, Jie; Ning, Jinying; Wery, Jean Pierre; Li, Qi-Xiang

    2016-07-01

    Anecdote clinical observations hint that non-small cell lung cancer (NSCLC) with exon-20 insertions might respond poorly to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), contrasting to those with classic mutations. Lack of patient-derived experimental models has been a major hurdle for the discovery of new treatment for the diseases. We established two NSCLC-PDXs harboring two different exon-20 insertions, LU0387-adenocarcinoma (ADC) with a nine-base insertion at 2319 (H773-V774insNPH) and LU3075-squamous cell carcinoma (SCC) with a nine-base insertion at 2316 (P772-H773insDNP). Both insertions immediately follow the regulatory C-helix of the kinase domain. Contrary to the generally good responses to EGFR inhibitors observed in PDXs with classic mutations, both exon-20 insertions are largely resistant to cetuximab and TKIs in vivo, suggesting fundamental difference from the classic EGFR mutations, consistent with the poor response rate to TKI seen in anecdotal clinic reports. It is worth noting that although responses are generally poor, they differ between the two exon-20 mutants depending on the type of TKI. In vitro drug sensitivity assays using established primary cell lines from our two PDXs largely confirmed the in vivo data. Our data from patient-derived experimental models confirmed that exon-20 insertions in domain immediately following the C-helix confer poor response to all known EGFR inhibitors, and suggested that these models can be utilized to facilitate the discovery of new therapies targeting NSCLC harboring exon-20 insertions. PMID:26891175

  2. Toxoplasma gondii Chitinase Induces Macrophage Activation.

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    Fausto Almeida

    Full Text Available Toxoplasma gondii is an obligate intracellular protozoan parasite found worldwide that is able to chronically infect almost all vertebrate species, especially birds and mammalians. Chitinases are essential to various biological processes, and some pathogens rely on chitinases for successful parasitization. Here, we purified and characterized a chitinase from T. gondii. The enzyme, provisionally named Tg_chitinase, has a molecular mass of 13.7 kDa and exhibits a Km of 0.34 mM and a Vmax of 2.64. The optimal environmental conditions for enzymatic function were at pH 4.0 and 50 °C. Tg_chitinase was immunolocalized in the cytoplasm of highly virulent T. gondii RH strain tachyzoites, mainly at the apical extremity. Tg_chitinase induced macrophage activation as manifested by the production of high levels of pro-inflammatory cytokines, a pathogenic hallmark of T. gondii infection. In conclusion, to our knowledge, we describe for the first time a chitinase of T. gondii tachyzoites and provide evidence that this enzyme might influence the pathogenesis of T. gondii infection.

  3. Association of kinase insert domain-containing receptor (KDR) gene polymorphism/ haplotypes with recurrent spontaneous abortion and genetic structure

    Science.gov (United States)

    Shahsavari, Shiva; Noormohammadi, Zahra; Zare Karizi, Shohreh

    2015-01-01

    Background: Recurrent spontaneous abortion is one of the diseases that can lead to physical, psychological, and, economical problems for both individuals and society. Recently a few numbers of genetic polymorphisms in kinase insert domain-containing receptor (KDR) gene are examined that can endanger the life of the fetus in pregnant women. Objective: The risk of KDR gene polymorphisms was investigated in Iranian women with idiopathic recurrent spontaneous abortion (RSA). Materials and Methods: A case controlled study was performed. One hundred idiopathic recurrent spontaneous abortion patients with at least two consecutive pregnancy losses before 20 weeks of gestational age with normal karyotypes were included in the study. Also, 100 healthy women with at least one natural pregnancy were studied as control group. Two functional SNPs located in KDR gene; rs1870377 (Q472H), and rs2305948 (V297I) as well as one tag SNP in the intron region (rs6838752) were genotyped by using PCR based restriction fragment length polymorphism (PCR-RFLP) technique. Haplotype frequency was determined for these three SNPs’ genotypes. Analysis of genetic STRUCTURE and K means clustering were performed to study genetic variation. Results: Functional SNP (rs1870377) was highly linked to tag SNP (rs6838752) (D´ value=0. 214; χ2 = 16.44, p<0. 001). K means clustering showed that k = 8 as the best fit for the optimal number of genetic subgroups in our studied materials. This result was in agreement with Neighbor Joining cluster analysis. Conclusion: In our study, the allele and genotype frequencies were not associated with RSA between patient and control individuals. Inconsistent results in different populations with different allele frequencies among RSA patients and controls may be due to ethnic variation and used sample size. PMID:27141535

  4. Pulmonary cryptococcosis induces chitinase in the rat

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    Casadevall Arturo

    2008-05-01

    Full Text Available Abstract Background We previously demonstrated that chronic pulmonary infection with Cryptococcus neoformans results in enhanced allergic inflammation and airway hyperreactivity in a rat model. Because the cell wall of C. neoformans consists of chitin, and since acidic mammalian chitinase (AMCase has recently been implicated as a novel mediator of asthma, we sought to determine whether such infection induces chitinase activity and expression of AMCase in the rat. Methods We utilized a previously-established model of chronic C. neoformans pulmonary infection in the rat to analyze the activity, expression and localization of AMCase. Results Our studies indicate that intratracheal inoculation of C. neoformans induces chitinase activity within the lung and bronchoalveolar lavage fluid of infected rats. Chitinase activity is also elicited by pulmonary infection with other fungi (e.g. C. albicans, but not by the inoculation of dead organisms. Enhanced chitinase activity reflects increased AMCase expression by airway epithelial cells and alveolar macrophages. Systemic cryptococcosis is not associated with increased pulmonary chitinase activity or AMCase expression. Conclusion Our findings indicate a possible link between respiratory fungal infections, including C. neoformans, and asthma through the induction of AMCase.

  5. Identification of a novel calcium binding motif based on the detection of sequence insertions in the animal peroxidase domain of bacterial proteins.

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    Saray Santamaría-Hernando

    Full Text Available Proteins of the animal heme peroxidase (ANP superfamily differ greatly in size since they have either one or two catalytic domains that match profile PS50292. The orf PP_2561 of Pseudomonas putida KT2440 that we have called PepA encodes a two-domain ANP. The alignment of these domains with those of PepA homologues revealed a variable number of insertions with the consensus G-x-D-G-x-x-[GN]-[TN]-x-D-D. This motif has also been detected in the structure of pseudopilin (pdb 3G20, where it was found to be involved in Ca(2+ coordination although a sequence analysis did not reveal the presence of any known calcium binding motifs in this protein. Isothermal titration calorimetry revealed that a peptide containing this consensus motif bound specifically calcium ions with affinities ranging between 33-79 µM depending on the pH. Microcalorimetric titrations of the purified N-terminal ANP-like domain of PepA revealed Ca(2+ binding with a K(D of 12 µM and stoichiometry of 1.25 calcium ions per protein monomer. This domain exhibited peroxidase activity after its reconstitution with heme. These data led to the definition of a novel calcium binding motif that we have termed PERCAL and which was abundantly present in animal peroxidase-like domains of bacterial proteins. Bacterial heme peroxidases thus possess two different types of calcium binding motifs, namely PERCAL and the related hemolysin type calcium binding motif, with the latter being located outside the catalytic domains and in their C-terminal end. A phylogenetic tree of ANP-like catalytic domains of bacterial proteins with PERCAL motifs, including single domain peroxidases, was divided into two major clusters, representing domains with and without PERCAL motif containing insertions. We have verified that the recently reported classification of bacterial heme peroxidases in two families (cd09819 and cd09821 is unrelated to these insertions. Sequences matching PERCAL were detected in all kingdoms of

  6. Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.

    Science.gov (United States)

    Aghazadeh, Rustam; Zamani, Mohammadreza; Motallebi, Mostafa; Moradyar, Mehdi; Moghadassi Jahromi, Zahra

    2016-09-01

    Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant. PMID:27430511

  7. WHEAT PATHOGEN RESISTANCE AND CHITINASE PROFILE

    Directory of Open Access Journals (Sweden)

    Zuzana Gregorová

    2015-02-01

    Full Text Available The powdery mildew and leaf rust caused by Blumeria graminis and Puccinia recondita (respectively are common diseases of wheat throughout the world. These fungal diseases greatly affect crop productivity. Incorporation of effective and durable disease resistance is an important breeding objective for wheat improvement. We have evaluated resistance of four bread wheat (Triticum aestivum and four spelt wheat (Triticum spelta cultivars. Chitinases occurrence as well as their activity was determined in leaf tissues. There was no correlation between resistance rating and activity of chitinase. The pattern of chitinases reveals four isoforms with different size in eight wheat cultivars. A detailed understanding of the molecular events that take place during a plant–pathogen interaction is an essential goal for disease control in the future.

  8. A simple, flexible and efficient PCR-fusion/Gateway cloning procedure for gene fusion, site-directed mutagenesis, short sequence insertion and domain deletions and swaps

    Directory of Open Access Journals (Sweden)

    Etchells J Peter

    2009-10-01

    Full Text Available Abstract Background The progress and completion of various plant genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating binary vectors containing: gene fusions, variants from site-directed mutagenesis, addition of protein tags together with domain swaps and deletions. Furthermore, efficient cloning procedures, ideally high throughput, are essential for pyramiding of multiple gene constructs. Results Here, we present a simple, flexible and efficient PCR-fusion/Gateway cloning procedure for construction of binary vectors for a range of gene fusions or variants with single or multiple nucleotide substitutions, short sequence insertions, domain deletions and swaps. Results from selected applications of the procedure which include ORF fusion, introduction of Cys>Ser mutations, insertion of StrepII tag sequence and domain swaps for Arabidopsis secondary cell wall AtCesA genes are demonstrated. Conclusion The PCR-fusion/Gateway cloning procedure described provides an elegant, simple and efficient solution for a wide range of diverse and complicated cloning tasks. Through streamlined cloning of sets of gene fusions and modification variants into binary vectors for systematic functional studies of gene families, our method allows for efficient utilization of the growing sequence and expression data.

  9. Heterologous expression of new antifungal chitinase from wheat.

    Science.gov (United States)

    Singh, Arpita; Kirubakaran, S Isaac; Sakthivel, N

    2007-11-01

    Chitinases (EC 3.2.1.14) have been grouped into seven classes (class I-VII) on the basis of their structural properties. Chitinases expressed during plant-microbe interaction are involved in defense responses of host plant against pathogens. In the present investigation, chitinase gene from wheat has been subcloned and overexpressed in Escherichia coli BL-21 (DE3). Molecular phylogeny analyses of wheat chitinase indicated that it belongs to an acidic form of class VII chitinase (glycosyl hydrolase family 19) and shows 77% identity with other wheat chitinase of class IV and low level identity to other plant chitinases. The three-dimensional structural model of wheat chitinase showed the presence of 10 alpha-helices, 3 beta-strands, 21 loop turns and the presence of 6 cysteine residues that are responsible for the formation of 3 disulphide bridges. The active site residues (Glu94 and Glu103) may be suggested for its antifungal activity. Expression of chitinase (33 kDa) was confirmed by SDS-PAGE and Western hybridization analyses. The yield of purified chitinase was 20 mg/L with chitinase activity of 1.9 U/mg. Purified chitinase exerted a broad-spectrum antifungal activity against Colletotrichum falcatum (red rot of sugarcane) Pestalotia theae (leaf spot of tea), Rhizoctonia solani (sheath blight of rice), Sarocladium oryzae (sheath rot of rice) Alternaria sp. (grain discoloration of rice) and Fusarium sp. (scab of rye). Due to its innate antifungal potential wheat chitinase can be used to enhance fungal-resistance in crop plants. PMID:17697785

  10. Two forms of the membrane-bound state of the first C2 domain (C2A) of synaptotagminⅠand calcium-triggered membrane insertion

    Institute of Scientific and Technical Information of China (English)

    HE Yuhong; LI Xianghui; WANG Fu; XUE Yi; SUI Senfang

    2003-01-01

    The synaptic vesicle protein synaptotagminⅠ (sytⅠ) is a vesicle trans membrane protein present in synaptic vesicles, which has been proposed as the Ca 2+ sensor that regulates secretion. The C2A domain is the membrane proximal part of its cytoplasmic domain. The interaction between C2A and lipid bilayer has be en considered to be essential for triggering neurotransmitter release. In the pr esent work, the measurements of membrane surface tension and surface concentrati on showed that the C2A domain of sytⅠexhibited two membrane-bound states: the s urface adsorption state and the membrane insertion state. The surface absorption state formed in a Ca2+-independent manner with lower affinity, while the membra ne insertion state formed with high affinity was only found in the presence of C a2+. Both the Ca2+-independent and Ca2+-dependent sytⅠ- membrane interactions r equired anionic phospholipids, such as phosphatidylserine (PS). When expressed i nto rat pheochromocytoma (PC12) cells and human embryonic kidney (HEK-293) cells , as demonstrated by immunofluorescence staining and subcellular fractionation, most of the C2A was found at the plasma membrane, even when the cells were deple ted of Ca2+ by incubation with EGTA. These results suggested a new molecular mec hanism of sytⅠas a Ca2+ sensor in membrane fusion. Ca2+-independent surface ads orption might attach sytⅠto the release site during the docking or priming step . When intracellular Ca2+ increased, sytⅠtriggered the neurotransmitter release following the Ca2+-dependent penetration into the target membrane.

  11. The Structure of Plasmodium falciparum Blood-Stage 6-Cys Protein Pf41 Reveals an Unexpected Intra-Domain Insertion Required for Pf12 Coordination.

    Directory of Open Access Journals (Sweden)

    Michelle L Parker

    Full Text Available Plasmodium falciparum is an apicomplexan parasite and the etiological agent of severe human malaria. The complex P. falciparum life cycle is supported by a diverse repertoire of surface proteins including the family of 6-Cys s48/45 antigens. Of these, Pf41 is localized to the surface of the blood-stage merozoite through its interaction with the glycophosphatidylinositol-anchored Pf12. Our recent structural characterization of Pf12 revealed two juxtaposed 6-Cys domains (D1 and D2. Pf41, however, contains an additional segment of 120 residues predicted to form a large spacer separating its two 6-Cys domains. To gain insight into the assembly mechanism and overall architecture of the Pf12-Pf41 complex, we first determined the 2.45 Å resolution crystal structure of Pf41 using zinc single-wavelength anomalous dispersion. Structural analysis revealed an unexpected domain organization where the Pf41 6-Cys domains are, in fact, intimately associated and the additional residues instead map predominately to an inserted domain-like region (ID located between two β-strands in D1. Notably, the ID is largely proteolyzed in the final structure suggesting inherent flexibility. To assess the contribution of the ID to complex formation, we engineered a form of Pf41 where the ID was replaced by a short glycine-serine linker and showed by isothermal titration calorimetry that binding to Pf12 was abrogated. Finally, protease protection assays showed that the proteolytic susceptibility of the ID was significantly reduced in the complex, consistent with the Pf41 ID directly engaging Pf12. Collectively, these data establish the architectural organization of Pf41 and define an essential role for the Pf41 ID in promoting assembly of the Pf12-Pf41 heterodimeric complex.

  12. Role of Chitin and Chitinase/Chitinase-Like Proteins in Inflammation, Tissue Remodeling, and Injury

    Science.gov (United States)

    Lee, Chun Geun; Da Silva, Carla A.; Dela Cruz, Charles S.; Ahangari, Farida; Ma, Bing; Kang, Min-Jong; He, Chuan-Hua; Takyar, Seyedtaghi; Elias, Jack A.

    2013-01-01

    The 18 glycosyl hydrolase family of chitinases is an ancient gene family that is widely expressed from prokaryotes to eukaryotes. In mammals, despite the absence of endogenous chitin, a number of chitinases and chitinase-like proteins (C/CLPs) have been identified. However, their roles have only recently begun to be elucidated. Acidic mammalian chitinase (AMCase) inhibits chitin-induced innate inflammation; augments chitin-free, allergen-induced Th2 inflammation; and mediates effector functions of IL-13. The CLPs BRP-39/YKL-40 (also termed chitinase 3-like 1) inhibit oxidant-induced lung injury, augments adaptive Th2 immunity, regulates apoptosis, stimulates alternative macrophage activation, and contributes to fibrosis and wound healing. In accord with these findings, levels of YKL-40 in the lung and serum are increased in asthma and other inflammatory and remodeling disorders and often correlate with disease severity. Our understanding of the roles of C/CLPs in inflammation, tissue remodeling, and tissue injury in health and disease is reviewed below. PMID:21054166

  13. Computational analysis of difenoconazole interaction with soil chitinases

    International Nuclear Information System (INIS)

    This study focusses on the investigation of the potential binding of the fungicide difenoconazole to soil chitinases using a computational approach. Computational characterization of the substrate binding sites of Serratia marcescens and Bacillus cereus chitinases using Fpocket tool reflects the role of hydrophobic residues for the substrate binding and the high local hydrophobic density of both sites. Molecular docking study reveals that difenoconazole is able to bind to Serratia marcescens and Bacillus cereus chitinases active sites, the binding energies being comparable

  14. WHEAT PATHOGEN RESISTANCE AND CHITINASE PROFILE

    OpenAIRE

    Zuzana Gregorová; Peter Socha; Marína Maglovski; Jana Moravčíková; Jana Libantová; Roman Kuna; Pavol Hauptvogel; Matušíková Ildikó

    2015-01-01

    The powdery mildew and leaf rust caused by Blumeria graminis and Puccinia recondita (respectively) are common diseases of wheat throughout the world. These fungal diseases greatly affect crop productivity. Incorporation of effective and durable disease resistance is an important breeding objective for wheat improvement. We have evaluated resistance of four bread wheat (Triticum aestivum) and four spelt wheat (Triticum spelta) cultivars. Chitinases occurrence as well as their activity was dete...

  15. The chitinase C gene PsChiC from Pseudomonas sp. and its synergistic effects on larvicidal activity

    Directory of Open Access Journals (Sweden)

    Wanfang Zhong

    2015-09-01

    Full Text Available Pseudomonas sp. strain TXG6-1, a chitinolytic gram-negative bacterium, was isolated from a vegetable field in Taixing city, Jiangsu Province, China. In this study, a Pseudomonas chitinase C gene (PsChiC was isolated from the chromosomal DNA of this bacterium using a pair of specific primers. The PsChiC gene consisted of an open reading frame of 1443 nucleotides and encoded 480 amino acid residues with a calculated molecular mass of 51.66 kDa. The deduced PsChiC amino acid sequence lacked a signal sequence and consisted of a glycoside hydrolase family 18 catalytic domain responsible for chitinase activity, a fibronectin type III-like domain (FLD and a C-terminal chitin-binding domain (ChBD. The amino acid sequence of PsChiCshowed high sequence homology (> 95% with chitinase C from Serratia marcescens. SDS-PAGE showed that the molecular mass of chitinase PsChiC was 52 kDa. Chitinase assays revealed that the chitobiosidase and endochitinase activities of PsChiCwere 51.6- and 84.1-fold higher than those of pET30a, respectively. Although PsChiC showed little insecticidal activity towards Spodoptera litura larvae, an insecticidal assay indicated that PsChiC increased the insecticidal toxicity of SpltNPV by 1.78-fold at 192 h and hastened death. These results suggest that PsChiC from Pseudomonas sp. could be useful in improving the pathogenicity of baculoviruses.

  16. Molecular cloning of class III chitinase gene from Avicennia marina and its expression analysis in response to cadmium and lead stress.

    Science.gov (United States)

    Wang, Li-Ying; Wang, You-Shao; Zhang, Jing-Ping; Gu, Ji-Dong

    2015-10-01

    Mangrove species have high tolerance to heavy metal pollution. Chitinases have been widely reported as defense proteins in response to heavy metal stress in terrestrial plants. In this study, a full-length cDNA sequence encoding an acidic and basic class III chitinase (AmCHI III) was cloned by using RT-PCR and RACE methods in Avicennia marina. AmCHI III mRNA expression in leaf of A. marina were investigated under Cd, Pb stresses on using real-time quantitative PCR. The deduced AmCHI III protein consists of 302 amino acids, including a signal putative peptide region, and a catalytic domain. Homology modeling of the catalytic domain revealed a typical molecular structure of class III plant chitinases. Results further demonstrated that the regulation of AmCHI III mRNA expression in leaves was strongly dependent on Cd, Pb stresses. AmCHI III mRNA expressions were significantly increased in response to Cd, Pb, and peaked at 7 days Cd-exposure, 7 days Pb-exposure, respectively. AmCHI III mRNA expression exhibited more sensitive to Pb stress than Cd stress. This work was the first time cloing chitinase from A. marina, and it brought evidence on chitinase gene involving in heavy metals (Cd(2+) and Pb(2+)) resistance or detoxification in plants. Further studies including the promoter and upstream regulation, gene over-expression and the response of mangrove chitinases to other stresses will shed more light on the role of chitinase in mangrove plants. PMID:26044930

  17. Two chitinase-like proteins abundantly accumulated in latex of mulberry show insecticidal activity

    Directory of Open Access Journals (Sweden)

    Komatsu Aino

    2010-01-01

    Full Text Available Abstract Background Plant latex is the cytoplasm of highly specialized cells known as laticifers, and is thought to have a critical role in defense against herbivorous insects. Proteins abundantly accumulated in latex might therefore be involved in the defense system. Results We purified latex abundant protein a and b (LA-a and LA-b from mulberry (Morus sp. and analyzed their properties. LA-a and LA-b have molecular masses of approximately 50 and 46 kDa, respectively, and are abundant in the soluble fraction of latex. Western blotting analysis suggested that they share sequence similarity with each other. The sequences of LA-a and LA-b, as determined by Edman degradation, showed chitin-binding domains of plant chitinases at the N termini. These proteins showed small but significant chitinase and chitosanase activities. Lectin RCA120 indicated that, unlike common plant chitinases, LA-a and LA-b are glycosylated. LA-a and LA-b showed insecticidal activities when fed to larvae of the model insect Drosophila melanogaster. Conclusions Our results suggest that the two LA proteins have a crucial role in defense against herbivorous insects, possibly by hydrolyzing their chitin.

  18. Intramolecular activation of a Ca(2+)-dependent protein kinase is disrupted by insertions in the tether that connects the calmodulin-like domain to the kinase

    Science.gov (United States)

    Vitart, V.; Christodoulou, J.; Huang, J. F.; Chazin, W. J.; Harper, J. F.; Evans, M. L. (Principal Investigator)

    2000-01-01

    Ca(2+)-dependent protein kinases (CDPK) have a calmodulin-like domain (CaM-LD) tethered to the C-terminal end of the kinase. Activation is proposed to involve intramolecular binding of the CaM-LD to a junction sequence that connects the CaM-LD to the kinase domain. Consistent with this model, a truncated CDPK (DeltaNC) in which the CaM-LD has been deleted can be activated in a bimolecular interaction with an isolated CaM-LD or calmodulin, similar to the activation of a calmodulin-dependent protein kinase (CaMK) by calmodulin. Here we provide genetic evidence that this bimolecular activation requires a nine-residue binding segment from F436 to I444 (numbers correspond to CPK-1 accession number L14771). Two mutations at either end of this core segment (F436/A and VI444/AA) severely disrupted bimolecular activation, whereas flanking mutations had only minor effects. Intramolecular activation of a full-length kinase was also disrupted by a VI444/AA mutation, but surprisingly not by a F436/A mutation (at the N-terminal end of the binding site). Interestingly, intramolecular but not bimolecular activation was disrupted by insertion mutations placed immediately downstream of I444. To show that mutant enzymes were not misfolded, latent kinase activity was stimulated through binding of an antijunction antibody. Results here support a model of intramolecular activation in which the tether (A445 to G455) that connects the CaM-LD to the kinase provides an important structural constraint and is not just a simple flexible connection.

  19. Multiple Mechanisms Contribute to the Cell Growth Defects Imparted by Human Telomerase Insertion in Fingers Domain Mutations Associated with Premature Aging Diseases.

    Science.gov (United States)

    Chu, Tsz Wai; MacNeil, Deanna Elise; Autexier, Chantal

    2016-04-15

    Normal human stem cells rely on low levels of active telomerase to sustain their high replicative requirements. Deficiency in telomere maintenance mechanisms leads to the development of premature aging diseases, such as dyskeratosis congenita and aplastic anemia. Mutations in the unique "insertion in fingers domain" (IFD) in the human telomerase reverse transcriptase catalytic subunit (hTERT) have previously been identified and shown to be associated with dyskeratosis congenita and aplastic anemia. However, little is known about the molecular mechanisms impacted by these IFD mutations. We performed comparative functional analyses of disease-associated IFD variants at the molecular and cellular levels. We report that hTERT-P721R- and hTERT-R811C-expressing cells exhibited growth defects likely due to impaired TPP1-mediated recruitment of these variant enzymes to telomeres. We showed that activity and processivity of hTERT-T726M failed to be stimulated by TPP1-POT1 overexpression and that dGTP usage by this variant was less efficient compared with the wild-type enzyme. hTERT-P785L-expressing cells did not show growth defects, and this variant likely confers cell survival through increased DNA synthesis and robust activity stimulation by TPP1-POT1. Altogether, our data suggest that multiple mechanisms contribute to cell growth defects conferred by the IFD variants. PMID:26887940

  20. Pyrrolo[3,2-d]pyrimidine Derivatives as Type II Kinase Insert Domain Receptor (KDR Inhibitors: CoMFA and CoMSIA Studies

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    Jia-Jie Zhang

    2012-02-01

    Full Text Available Kinase insert domain receptor (KDR inhibitors have been proved to be very effective anticancer agents. Molecular docking, 3D-QSAR methods, CoMFA and CoMSIA were performed on pyrrolo[3,2-d]pyrimidine derivatives as non-ATP competitive KDR inhibitors (type II. The bioactive conformation was explored by docking one potent compound 20 into the active site of KDR in its DFG-out inactive conformation. The constructed CoMFA and CoMSIA models produced statistically significant results with the cross-validated correlation coefficients q2 of 0.542 and 0.552, non-cross-validated correlation coefficients r2 of 0.912 and 0.955, and predicted correction coefficients r2pred of 0.913 and 0.897, respectively. These results ensure the CoMFA and CoMSIA models as a tool to guide the design of a series of new potent KDR inhibitors.

  1. The -271 G>A polymorphism of kinase insert domain-containing receptor gene regulates its transcription level in patients with non-small cell lung cancer

    International Nuclear Information System (INIS)

    Kinase insert domain-containing receptor (KDR) plays a critical role in the metastasis of cancer and is used as a molecular target in cancer therapy. We investigated the characteristics of the -271 G>A polymorphism of the KDR gene to gain information that may benefit the development of individualized therapies for patients with non-small cell lung cancer (NSCLC). The -271 G>A polymorphism of the KDR gene in 106 lung cancer patients and 203 healthy control individuals was analyzed by polymerase chain reaction (PCR) and DNA sequencing methods. Real-time quantitative PCR and immunohistochemical methods were used to evaluate KDR mRNA and protein expression levels, respectively, in frozen tumor specimens. The -271 G>A polymorphism was associated with the mRNA expression level of the KDR gene in tumor tissues (t = 2.178, P = 0.032, independent samples t-test). Compared with the AG/GG genotype, the AA genotype was associated with higher KDR mRNA expression in tumor tissues. We found no relationship between the genotype and the KDR protein expression level and no significant difference in the distribution of the KDR gene polymorphism genotypes between lung cancer patients and the control group (χ2 = 1.269, P = 0.264, Fisher's exact test). This study is the first to show that the -271 G>A polymorphism of the KDR gene may be a functional polymorphism related to the regulation of gene transcription. These findings may have important implications for therapies targeting KDR in patients with NSCLC

  2. From bacteria to human: a journey into the world of chitinases.

    Science.gov (United States)

    Adrangi, Sina; Faramarzi, Mohammad Ali

    2013-12-01

    Chitinases, the enzymes responsible for the biological degradation of chitin, are found in a wide range of organisms from bacteria to higher plants and animals. They participate in numerous physiological processes such as nutrition, parasitism, morphogenesis and immunity. Many organisms, in addition to chitinases, produce inactive chitinase-like lectins that despite lacking enzymatic activity are involved in several regulatory functions. Most known chitinases belong to families 18 and 19 of glycosyl hydrolases, however a few chitinases that belong to families 23 and 48 have also been identified in recent years. In this review, different aspects of chitinases and chi-lectins from bacteria, fungi, insects, plants and mammals are discussed. PMID:24095741

  3. Preparation of chitooligosaccharides from fungal waste mycelium by recombinant chitinase.

    Science.gov (United States)

    Lv, Mengyuan; Hu, Ying; Gänzle, Michael G; Lin, Jianguo; Wang, Changgao; Cai, Jun

    2016-07-22

    This study aimed to develop an enzymatic method for conversion of chitin from fungal waste mycelia to chitooligosaccharides. The recombinant chitinase LlChi18A from Lactococcus lactis was over-expressed by Escherichia coli BL21 (DE3) and purified by affinity chromatography. The enzymatic properties of the purified enzyme were studied by chitin oligosaccharides. Waste mycelium was pre-treated by alkaline. The optimal conditions for hydrolysis of fungal chitin by recombinant chitinase were determined by Schales method. HPLC/ESI-MS was used to determine the content of N-acetylglucosamine and chitooligosaccharides after hydrolysis. The level of reducing sugar released from pretreated mycelium by chitinase increased with the reaction time during 6 days. The main product in the hydrolysates was N,N'-diacetylchitobiose. After hydrolysis by chitinase for 5 d, the yield of N,N'-diacetylchitobiose from waste mycelium was around 10% with estimated purity of around 70%. Combination of chitinase and snailase remarkably increased the yield to 24% with purity of 78%. Fungal mycelium which contains chitin is a new potential source for obtaining food grade chitooligosaccharides. PMID:27153004

  4. Transgenic expression of plant chitinases to enhance disease resistance.

    Science.gov (United States)

    Cletus, Jean; Balasubramanian, Vaiyapuri; Vashisht, Divya; Sakthivel, Natarajan

    2013-11-01

    Crop plants have evolved an array of mechanisms to counter biotic and abiotic stresses. Many pathogenesis-related proteins are expressed by plants during the attack of pathogens. Advances in recombinant DNA technology and understanding of plant-microbe interactions at the molecular level have paved the way for isolation and characterization of genes encoding such proteins, including chitinases. Chitinases are included in families 18 and 19 of glycosyl hydrolases (according to www.cazy.org ) and they are further categorized into seven major classes based on their aminoacid sequence homology, three-dimensional structures, and hydrolytic mechanisms of catalytic reactions. Although chitin is not a component of plant cell walls, plant chitinases are involved in development and non-specific stress responses. Also, chitinase genes sourced from plants have been successfully over-expressed in crop plants to combat fungal pathogens. Crops such as tomato, potato, maize, groundnut, mustard, finger millet, cotton, lychee, banana, grape, wheat and rice have been successfully engineered for fungal resistance either with chitinase alone or in combination with other PR proteins. PMID:23794096

  5. Induction and purification of chitinase in Brassica napus L. ssp. oleifera infected with Phoma lingam

    DEFF Research Database (Denmark)

    Rasmussen, U.; Giese, H.; Dalgaard Mikkelsen, J.

    1992-01-01

    after tryptic digestion of the protein shows high sequence similarity to basic chitinases from bean, tobacco, potato, Arabidopsis, barley and rice, as well as to acidic chitinases from tobacco and petunia. A close serological relationship was found between the chitinase isoenzyme and an isoenzyme from...

  6. Calorimeter insertion

    CERN Multimedia

    2006-01-01

    Calorimeter insertion between toroids in the ATLAS experiment detector Calorimeters are surrounding the inner detector. Calorimeters will absorb and measure the energies of the most charged and neutral particles after the collisions. The saved energy in the calorimeter is detected and converted to signals that are taken out with data taking electronics.

  7. Cloning, expression and biocharacterization of OfCht5, the chitinase from the insect Ostrinia furnacalis

    Institute of Scientific and Technical Information of China (English)

    Qingyue Wu; Tian Liu; Qing Yang

    2013-01-01

    Chitinase catalyzes β-l,4-glycosidic linkages in chitin and has attracted research interest due to it being a potential pesticide target and an enzymatic tool for preparation of N-acetyl-β-D-glucosamine.An individual insect contains multiple genes encoding chitinases,which vary in domain architectures,expression patterns,physiological roles and biochemical properties.Herein,OfCht5,the glycoside hydrolase family 18 chitinase from the widespread lepidopteran pest Ostrinia furnacalis,was cloned,expressed in the yeast Pichia pastoris and biochemically characterized in an attempt to facilitate both pest control and biomaterial preparation.Complementary DNA sequence analysis indicated that OfCHT5 consisted of an open reading frame of 1 665-bp nucleotides.Phylogenic analysis suggested OfCht5 belongs to the Group Ⅰ insect chitinases.Expression of OfCht5 in Pichia pastoris resulted in highest specific activity after 120 h of induction with methanol.Through two steps of purification,consisting of ammonium sulfate precipitation and metal chelating chromatography,about 7 mg of the recombinant OfCht5 was purified to homogeneity from 1 L culture supematant.OfCht5 effectively converted colloidal chitin into chitobiose,but had relatively low activity toward α-chitin.When chitooligosaccharides [(GlcNAc)n,n =3-6] were used as substrates,OfCht5 was observed to possess the highest catalytic efficiency parameter toward (GlcNAc)4 and predominantely hydrolyzed the second glycosidic bond from the non-reducing end.Together with β-N-acetyl-D-hexosaminidase OfHexl,OfCht5 achieved its highest efficiency in chitin degradation that yielded N-acetyl-β-D-glucosamine,a valuable pharmacological reagent and food supplement,within a molar concentration ratio of OfCht5 versus OfHexl in the range of 9∶1-15 ∶ 1.This work provides an alternative to existing preparation ofchitinase for pesticides and other applications.

  8. Insertion sequences.

    OpenAIRE

    Mahillon, Jacques; Chandler, M.

    1998-01-01

    Insertion sequences (ISs) constitute an important component of most bacterial genomes. Over 500 individual ISs have been described in the literature to date, and many more are being discovered in the ongoing prokaryotic and eukaryotic genome-sequencing projects. The last 10 years have also seen some striking advances in our understanding of the transposition process itself. Not least of these has been the development of various in vitro transposition systems for both prokaryotic and eukaryoti...

  9. A single base insertion in the putative transmembrane domain of the tyrosinase gene as a cause for tyrosinase-negative oculocutaneous albinism

    Energy Technology Data Exchange (ETDEWEB)

    Chintamaneni, C.D.; Kobayashi, Y.; Kwon, B.S. (Indiana Univ. School of Medicine, Indianapolis (United States)); Halaban, R. (Yale Univ. School of Medicine, New Haven, CT (United States)); Witkop, C.J. Jr. (Univ. of Minnesota, Minneapolis (United States))

    1991-06-15

    The authors have determined a molecular defect to be the likely basis for inactivity of the tyrosinase from a patient with tyrosinase-negative oculocutaneous albinism. A single base (thymine) was inserted in exon 5 of the tyrosinase gene following codon 471 in the putative transmembrane coding region. This insertion caused a shift in the reading frame of 19 amino acids at the 3{prime} end and introduced a premature termination signal that would be expected to truncate the protein by 21 amino acids at the carboxyl terminus. The albino tyrosinase was not recognized by antibodies directed to the carboxyl terminus of tyrosinase. Furthermore, as shown by gel electrophoresis of the immunoprecipitated protein, the tyrosinase was {approx} 3kDa smaller than normal. Similar immunoprecipitation data were obtained when cloned normal and mutant tyrosinases were expressed in COS-1 cells.

  10. Nitrogen regulates chitinase gene expression in a marine bacterium

    DEFF Research Database (Denmark)

    Delpin, Marina; Goodman, A.E.

    2009-01-01

    Ammonium concentration and nitrogen source regulate promoter activity and use for the transcription of chiA, the major chitinase gene of Pseudoalteromonas sp. S91 and S91CX, an S91 transposon lacZ fusion mutant. The activity of chiA was quantified by beta-galactosidase assay of S91CX cultures con...

  11. Chitinases from Bacteria to Human: Properties, Applications, and Future Perspectives

    Directory of Open Access Journals (Sweden)

    Abhishek Singh Rathore

    2015-01-01

    Full Text Available Chitin is the second most plenteous polysaccharide in nature after cellulose, present in cell walls of several fungi, exoskeletons of insects, and crustacean shells. Chitin does not accumulate in the environment due to presence of bacterial chitinases, despite its abundance. These enzymes are able to degrade chitin present in the cell walls of fungi as well as the exoskeletons of insect. They have shown being the potential agents for biological control of the plant diseases caused by various pathogenic fungi and insect pests and thus can be used as an alternative to chemical pesticides. There has been steady increase in demand of chitin derivatives, obtained by action of chitinases on chitin polymer for various industrial, clinical, and pharmaceutical purposes. Hence, this review focuses on properties and applications of chitinases starting from bacteria, followed by fungi, insects, plants, and vertebrates. Designing of chitinase by applying directed laboratory evolution and rational approaches for improved catalytic activity for cost-effective field applications has also been explored.

  12. Stability of transgene integration and expression in subsequent generations of doubled haploid oilseed rape transformed with chitinase and beta-1,3-glucanase genes in a double-gene construct.

    Science.gov (United States)

    Melander, Margareta; Kamnert, Iréne; Happstadius, Ingrid; Liljeroth, Erland; Bryngelsson, Tomas

    2006-09-01

    A double-gene construct with one chitinase and one beta-1,3-glucanase gene from barley, both driven by enhanced 35S promoters, was transformed into oilseed rape. From six primary transformants expressing both transgenes 10 doubled haploid lines were produced and studied for five generations. The number of inserted copies for both the genes was determined by Southern blotting and real-time PCR with full agreement between the two methods. When copy numbers were analysed in different generations, discrepancies were found, indicating that at least part of the inserted sequences were lost in one of the alleles of some doubled haploids. Chitinase and beta-1,3-glucanase expression was analysed by Western blotting in all five doubled haploid generations. Despite that both the genes were present on the same T-DNA and directed by the same promoter their expression pattern between generations was different. The beta-1,3-glucanase was expressed at high and stable levels in all generations, while the chitinase displayed lower expression that varied between generations. The transgenic plants did not show any major impact on fungal resistance when assayed in greenhouse, although purified beta-1,3-glucanase and chitinase caused retardment of fungal growth in vitro. PMID:16565860

  13. Insertional Mutations in Herpes Simplex Virus Type 1 gL Identify Functional Domains for Association with gH and for Membrane Fusion▿

    OpenAIRE

    Fan, Qing; Lin, Erick; Spear, Patricia G.

    2009-01-01

    Glycoprotein L (gL) is one of four glycoproteins required for the entry of herpes simplex virus (HSV) into cells and for virus-induced cell fusion. This glycoprotein oligomerizes with gH to form a membrane-bound heterodimer but can be secreted when expressed without gH. Twelve unique gL linker-insertion mutants were generated to identify regions critical for gH binding and gH/gL processing and regions essential for cell fusion and viral entry. All gL mutants were detected on the cell surface ...

  14. Bipartite Topology of Treponema pallidum Repeat Proteins C/D and I: OUTER MEMBRANE INSERTION, TRIMERIZATION, AND PORIN FUNCTION REQUIRE A C-TERMINAL β-BARREL DOMAIN.

    Science.gov (United States)

    Anand, Arvind; LeDoyt, Morgan; Karanian, Carson; Luthra, Amit; Koszelak-Rosenblum, Mary; Malkowski, Michael G; Puthenveetil, Robbins; Vinogradova, Olga; Radolf, Justin D

    2015-05-01

    We previously identified Treponema pallidum repeat proteins TprC/D, TprF, and TprI as candidate outer membrane proteins (OMPs) and subsequently demonstrated that TprC is not only a rare OMP but also forms trimers and has porin activity. We also reported that TprC contains N- and C-terminal domains (TprC(N) and TprC(C)) orthologous to regions in the major outer sheath protein (MOSP(N) and MOSP(C)) of Treponema denticola and that TprC(C) is solely responsible for β-barrel formation, trimerization, and porin function by the full-length protein. Herein, we show that TprI also possesses bipartite architecture, trimeric structure, and porin function and that the MOSP(C)-like domains of native TprC and TprI are surface-exposed in T. pallidum, whereas their MOSP(N)-like domains are tethered within the periplasm. TprF, which does not contain a MOSP(C)-like domain, lacks amphiphilicity and porin activity, adopts an extended inflexible structure, and, in T. pallidum, is tightly bound to the protoplasmic cylinder. By thermal denaturation, the MOSP(N) and MOSP(C)-like domains of TprC and TprI are highly thermostable, endowing the full-length proteins with impressive conformational stability. When expressed in Escherichia coli with PelB signal sequences, TprC and TprI localize to the outer membrane, adopting bipartite topologies, whereas TprF is periplasmic. We propose that the MOSP(N)-like domains enhance the structural integrity of the cell envelope by anchoring the β-barrels within the periplasm. In addition to being bona fide T. pallidum rare outer membrane proteins, TprC/D and TprI represent a new class of dual function, bipartite bacterial OMP. PMID:25805501

  15. Molecular phylogeny of C1 inhibitor depicts two immunoglobulin-like domains fusion in fishes and ray-finned fishes specific intron insertion after separation from zebrafish

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Abhishek, E-mail: akumar@bot.uni-kiel.de [Department of Genetics and Molecular Biology in Botany, Institute of Botany, Christian-Albrechts-University at Kiel, Kiel (Germany); Bhandari, Anita [Molecular Physiology, Zoological Institute, Christian-Albrechts-University at Kiel, Kiel (Germany); Sarde, Sandeep J. [Department of Genetics and Molecular Biology in Botany, Institute of Botany, Christian-Albrechts-University at Kiel, Kiel (Germany); Goswami, Chandan [National Institute of Science Education and Research, Bhubaneswar, Orissa (India)

    2014-07-18

    Highlights: • C1 inhibitors of fishes have two Ig domains fused in the N-terminal end. • Spliceosomal introns gain in two Ig domains of selected ray-finned fishes. • C1 inhibitors gene is maintained from 450 MY on the same locus. • C1 inhibitors gene is missing in frog and lampreys. • C1 inhibitors of tetrapod and fishes differ in the RCL region. - Abstract: C1 inhibitor (C1IN) is a multi-facet serine protease inhibitor in the plasma cascades, inhibiting several proteases, notably, regulates both complement and contact system activation. Despite huge advancements in the understanding of C1IN based on biochemical properties and its roles in the plasma cascades, the phylogenetic history of C1IN remains uncharacterized. To date, there is no comprehensive study illustrating the phylogenetic history of C1IN. Herein, we explored phylogenetic history of C1IN gene in vertebrates. Fishes have C1IN with two immunoglobulin like domains attached in the N-terminal region. The RCL regions of CIIN from fishes and tetrapod genomes have variations at the positions P2 and P1′. Gene structures of C1IN gene from selected ray-finned fishes varied in the Ig domain region with creation of novel intron splitting exon Im2 into Im2a and Im2b. This intron is limited to ray-finned fishes with genome size reduced below 1 Gb. Hence, we suggest that genome compaction and associated double-strand break repairs are behind this intron gain. This study reveals the evolutionary history of C1IN and confirmed that this gene remains the same locus for ∼450 MY in 52 vertebrates analysed, but it is not found in frogs and lampreys.

  16. Molecular phylogeny of C1 inhibitor depicts two immunoglobulin-like domains fusion in fishes and ray-finned fishes specific intron insertion after separation from zebrafish

    International Nuclear Information System (INIS)

    Highlights: • C1 inhibitors of fishes have two Ig domains fused in the N-terminal end. • Spliceosomal introns gain in two Ig domains of selected ray-finned fishes. • C1 inhibitors gene is maintained from 450 MY on the same locus. • C1 inhibitors gene is missing in frog and lampreys. • C1 inhibitors of tetrapod and fishes differ in the RCL region. - Abstract: C1 inhibitor (C1IN) is a multi-facet serine protease inhibitor in the plasma cascades, inhibiting several proteases, notably, regulates both complement and contact system activation. Despite huge advancements in the understanding of C1IN based on biochemical properties and its roles in the plasma cascades, the phylogenetic history of C1IN remains uncharacterized. To date, there is no comprehensive study illustrating the phylogenetic history of C1IN. Herein, we explored phylogenetic history of C1IN gene in vertebrates. Fishes have C1IN with two immunoglobulin like domains attached in the N-terminal region. The RCL regions of CIIN from fishes and tetrapod genomes have variations at the positions P2 and P1′. Gene structures of C1IN gene from selected ray-finned fishes varied in the Ig domain region with creation of novel intron splitting exon Im2 into Im2a and Im2b. This intron is limited to ray-finned fishes with genome size reduced below 1 Gb. Hence, we suggest that genome compaction and associated double-strand break repairs are behind this intron gain. This study reveals the evolutionary history of C1IN and confirmed that this gene remains the same locus for ∼450 MY in 52 vertebrates analysed, but it is not found in frogs and lampreys

  17. IN SILICO ANALYSIS OF CHITINASE PROMOTER ISOLATED FROM DROSERA ROTUNDIFOLIA L.

    OpenAIRE

    Dominika Ďurechová; Ildikó Matušíková; Jana Moravčíková; Martin Jopčík; Jana Libantová

    2014-01-01

    Chitinases occur in dozens of genes in the individual plant species and play diverse roles in plant growth and development, and during the plant defense to biotic and abiotic stress. Here we focused on isolation and in silico characterization of regulatory sequences of chitinase gene that belongs to the first isolated gene sequences from D. rotundifolia overall. For the isolation of the 739 bp sequence of chitinase promoter the genome walking approach was applied. The authenticity of the obta...

  18. Reduced Chitinase Activities in Ant Plants of the Genus Macaranga

    Science.gov (United States)

    Heil, Martin; Fiala, Brigitte; Linsenmair, K. Eduard; Boller, Thomas

    Many plant species have evolved mutualistic associations with ants, protecting their host against detrimental influences such as herbivorous insects. Letourneau (1998) reported in the case of Piper that ants defend their plants principally against stem-boring insects and also reduce fungal infections on inflorescences. Macaranga plants that were experimentally deprived of their symbiotic Crematogaster ants suffered heavily from shoot borers and pathogenic fungi (Heil 1998). Here we report that ants seem to reduce fungal infections actively in the obligate myrmecophyte Macarangatriloba (Euphorbiaceae), while ant-free plants can be easily infected. We also found extremely low chitinase activity in Macaranga plants. The plants' own biochemical defense seems to be reduced, and low chitinase activity perhaps may represent a predisposition for the evolution of myrmecophytism. These plants are therefore highly dependent on their ants, which obviously function not only as an antiherbivore defense but also as an effective agent against fungal pathogens.

  19. EXPRESSION OF CHITINASE GENE IN TRANSGENIC RAPE PLANTS

    OpenAIRE

    Lu Longdou; Gao Wu Jun; Wang Jingxue; Duan Hongying; Li Ruili

    2005-01-01

    The hypocotyl and cotyledon of Brassica napus L. H165 and Brassica juncea DB3 were transformed with chitinase gene and herbicide-resistance gene by co-culture with Agrobacterium tumefacients LBA4404, and rape plants were obtained which could grow on the medium containing herbicide. The PCR result showed that exotic genes were integrated in the genome of the rape. Further study was performed to determine the impact of temperature on the transgenic rate and the differentiation o...

  20. Isolation of bacteria producing chitinase and inhibiting growth of Rhizoctonia solani

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ Five bacteria strains with higher chitinase activity were isolated by using a technique of enriched cell wall of R. solani. All of them showed inhibiting effect on the growth of R. solani. Being cultured 3 d, strain CH-1 showed higher chitinase activity on the chitin plate. The diameter of the transparent circle reached 8.7 mm (4 replications) . In the antagonistic test to R. solani in PDA plate, the circle was 18.1 mm. It was also observed that the antagonistic ability of some strains was not consistent with the chitinase activity (Table 1). It may be connected with the secretion of chitinase at different culture situations.

  1. Molecular and functional evolution of class I chitinases for plant carnivory in the caryophyllales.

    Science.gov (United States)

    Renner, Tanya; Specht, Chelsea D

    2012-10-01

    Proteins produced by the large and diverse chitinase gene family are involved in the hydrolyzation of glycosidic bonds in chitin, a polymer of N-acetylglucosamines. In flowering plants, class I chitinases are important pathogenesis-related proteins, functioning in the determent of herbivory and pathogen attack by acting on insect exoskeletons and fungal cell walls. Within the carnivorous plants, two subclasses of class I chitinases have been identified to play a role in the digestion of prey. Members of these two subclasses, depending on the presence or absence of a C-terminal extension, can be secreted from specialized digestive glands found within the morphologically diverse traps that develop from carnivorous plant leaves. The degree of homology among carnivorous plant class I chitinases and the method by which these enzymes have been adapted for the carnivorous habit has yet to be elucidated. This study focuses on understanding the evolution of carnivory and chitinase genes in one of the major groups of plants that has evolved the carnivorous habit: the Caryophyllales. We recover novel class I chitinase homologs from species of genera Ancistrocladus, Dionaea, Drosera, Nepenthes, and Triphyophyllum, while also confirming the presence of two subclasses of class I chitinases based upon sequence homology and phylogenetic affinity to class I chitinases available from sequenced angiosperm genomes. We further detect residues under positive selection and reveal substitutions specific to carnivorous plant class I chitinases. These substitutions may confer functional differences as indicated by protein structure homology modeling. PMID:22490823

  2. Purification and characterization of chitinase from Streptomyces violascens NRRL B2700.

    Science.gov (United States)

    Gangwar, Mamta; Singh, Vineeta; Pandey, Asheesh Kumar; Tripathi, C K M; Mishra, B N

    2016-01-01

    Chitinase is one of the important enzymes as it is directly linked to Chitin that has wide applications in industrial, medical and commercial fields for its biocompatibility and biodegradability. Here, we report extracellular chitinase production by Streptomyces violascens NRRL B2700 under submerged fermentation condition. Chitinase production started after 10 h of incubation and reached to maximum level at 72 h of cultivation. Studies on the influence of additional carbon and nitrogen sources on chitinase production revealed that maltose, xylose, fructose, lactose, soybean meal and ammonium nitrate served as good carbon and nitrogen sources to enhance chitinase yield by 1.6 to 6 fold. Medium supplemented with 1% colloidal chitin produced high chitinase concentration (0.1714 U/mg). The enzyme chitinase was purified from the culture broth by 75% ammonium sulphate precipitation, DEAE-cellulose ion-exchange and sephadex G-100 gel filtration. The molecular mass of the purified chitinase was 65 kDa as estimated by SDS-PAGE. The apparent Michaelis constant (K(m)) and the maximum rate (V(max)) of the enzyme for colloidal chitin were 1.556 mg/mL and 2.680 μM/min/mg, respectively suggested high affinity towards-chitin. Possibly, it is the first report on production of chitinase from S. violascens NRRL B2700. The findings were encouraging, especially for cost effective production, and further warrants media and purification optimization studies for enhanced yield. PMID:26891554

  3. ZYMOGRAPHIC IDENTIFICATION AND BIOCHEMICAL CHARACTERIZATION OF CHITINASE AGAINST PHYTOFUNGAL PATHOGENS

    Directory of Open Access Journals (Sweden)

    Urja Pandya

    2014-08-01

    Full Text Available An endospore forming Gram positive bacterium (MBCU4 was isolated from a vermicompost amended soil, and confirmed as Bacillus subtilis through the 16S rRNA sequence analysis. An extracellular chitinase was detected from this strain of B. subtilis under specific environmental condition. An attempt was made to purify the enzyme by ammonium sulfate precipitation followed by DEAE sepharose CL-6B column chromatography. The purified enzyme was demonstrated as a single band, having the molecular weight 31kDa on SDS PAGE analysis and its activity in the gel was determined by clear zone on zymogram. Further characterization of the isolated enzymes has showed that this enzyme is most active at pH 6.0 and at the optimized temperature of 50 0C. The purified chitinase exhibited high degree of antifungal activity particularly by degrading their cell wall components of plant pathogens Macrophomina phaseolina (69.0% and Rhizoctonia solani (52.0%. It infers that the chitinase produced by B. subtilis could play an important role for biopesticidal activity.

  4. Insertions and the emergence of novel protein structure: a structure-based phylogenetic study of insertions

    Directory of Open Access Journals (Sweden)

    Blouin Christian

    2007-11-01

    Full Text Available Abstract Background In protein evolution, the mechanism of the emergence of novel protein domain is still an open question. The incremental growth of protein variable regions, which was produced by stochastic insertions, has the potential to generate large and complex sub-structures. In this study, a deterministic methodology is proposed to reconstruct phylogenies from protein structures, and to infer insertion events in protein evolution. The analysis was performed on a broad range of SCOP domain families. Results Phylogenies were reconstructed from protein 3D structural data. The phylogenetic trees were used to infer ancestral structures with a consensus method. From these ancestral reconstructions, 42.7% of the observed insertions are nested insertions, which locate in previous insert regions. The average size of inserts tends to increase with the insert rank or total number of insertions in the variable regions. We found that the structures of some nested inserts show complex or even domain-like fold patterns with helices, strands and loops. Furthermore, a basal level of structural innovation was found in inserts which displayed a significant structural similarity exclusively to themselves. The β-Lactamase/D-ala carboxypeptidase domain family is provided as an example to illustrate the inference of insertion events, and how the incremental growth of a variable region is capable to generate novel structural patterns. Conclusion Using 3D data, we proposed a method to reconstruct phylogenies. We applied the method to reconstruct the sequences of insertion events leading to the emergence of potentially novel structural elements within existing protein domains. The results suggest that structural innovation is possible via the stochastic process of insertions and rapid evolution within variable regions where inserts tend to be nested. We also demonstrate that the structure-based phylogeny enables the study of new questions relating to the

  5. Entamoeba histolytica Lectins Contain Unique 6-Cys or 8-Cys Chitin-Binding Domains

    OpenAIRE

    Van Dellen, Katrina; Ghosh, Sudip K.; Robbins, Phillips W.; Loftus, Brendan; Samuelson, John

    2002-01-01

    The Jacob lectin, the most abundant glycoprotein in the cyst wall of Entamoeba invadens, contains five unique 6-Cys chitin-binding domains (CBDs). We identified Entamoeba histolytica and Entamoeba dispar genes encoding Jacob homologues, each of which contains two predicted 6-Cys CBDs. A unique 8-Cys CBD was found at the N termini of the E. histolytica chitinase and three other predicted lectins, called Jessie 1 to Jessie 3. The CBDs of four E. histolytica lectins (Jacob, chitinase, and Jessie...

  6. Polymorphisms and Haplotypes of Acid Mammalian Chitinase Are Associated with Bronchial Asthma

    OpenAIRE

    Bierbaum, Sibylle; Nickel, Renate; Koch, Anja; Lau, Susanne; Deichmann, Klaus A.; Wahn, Ulrich; Superti-Furga, Andrea; Heinzmann, Andrea

    2005-01-01

    Rationale: Chitinases are enzymes that cleave chitin, a polysaccharide contained in many parasites of humans. Recent studies in mouse models of bronchial asthma have shown that acid mammalian chitinase (AMCase) is involved in the pathophysiology of asthma. It acts downstream of interleukin-13; inhibition of AMCase leads to an abrogated T-helper cell 2 inflammation, less bronchial hyperreactivity, and fewer eosinophils.

  7. Chitinase Expression in Listeria monocytogenes Is Positively Regulated by the Agr System

    DEFF Research Database (Denmark)

    Paspaliari, Dafni Katerina; Mollerup, Maria Storm; Kallipolitis, Birgitte H.;

    2014-01-01

    The food-borne pathogen Listeria monocytogenes encodes two chitinases, ChiA and ChiB, which allow the bacterium to hydrolyze chitin, the second most abundant polysaccharide in nature. Intriguingly, despite the absence of chitin in human and mammalian hosts, both of the chitinases have been deemed...

  8. Structural and functional analysis of chitinase gene family in wheat (Triticum aestivum).

    Science.gov (United States)

    Mishra, A K; Pandey, Bharati; Tyagi, Chetna; Chakraborty, Ohika; Kumar, Amrender; Jain, A K

    2015-04-01

    Chitinases are the hydrolytic enzymes which protect plants against pathogen attack. However, the precise role of chitinases in disease resistance has not been explored in wheat. In the present study, in silico approach, including secondary structure analysis, detailed signature pattern study, cis-acting regulatory elements survey, evolutionary trends and three-dimensional molecular modeling was used for different chitinase classes of wheat (Triticum aestivum). Homology modeling of class I, II, IV and 3 chitinase proteins was performed using the template crystal structure. The model structures were further refined by molecular mechanics methods using different tools, such as Procheck, ProSA and Verify3D. Secondary structure studies revealed greater percentage of residues forming a helix conformation with specific signature pattern, similar to casein kinase II phosphorylation site, amidation site, N-myristoylation (N-MYR) site and protein kinase C phoshorylation site. The expression profile suggested that wheat chitinase gene was highly expressed in cell culture and callus. We found that wheat chitinases showed more functional similarity with rice and barley. The results provide insight into the evolution of the chitinase family, constituting a diverse array of pathogenesis-related proteins. The study also provides insight into the possible binding sites of chitinase proteins and may further enhance our knowledge of fungal resistance mechanism in plants. PMID:26118129

  9. A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis

    DEFF Research Database (Denmark)

    Ohnuma, Takayuki; Numata, Tomoyuki; Osawa, Takuo;

    2011-01-01

    Expression of a class V chitinase gene (At4g19810, AtChiC) in Arabidopsis thaliana was examined by quantitative real-time PCR and by analyzing microarray data available at Genevestigator. The gene expression was induced by the plant stress-related hormones abscisic acid (ABA) and jasmonic acid (JA...... common to class V chitinases from higher plants....

  10. Amino-terminal sequence analysis of the Coccidioides immitis chitinase/immunodiffusion-complement fixation protein.

    OpenAIRE

    Johnson, S M; Zimmermann, C R; Pappagianis, D

    1993-01-01

    A chitinase isolated from Coccidioides immitis was subjected to amino-terminal protein sequence analysis. The resulting 18-amino-acid sequence was compared with the previously reported amino acid sequence of coccidioidal immunodiffusion-complement fixation (IDCF) antigen. From the homology of the two sequences, the results support the identification of the IDCF antigen with a chitinase.

  11. A Flip Turn for Membrane Protein Insertion

    OpenAIRE

    Shao, Sichen; Hegde, Ramanujan S.

    2011-01-01

    The transmembrane domains in a membrane protein must be recognized and correctly oriented before their insertion into the lipid bilayer. Devaraneni et al. (2011) generate snapshots at different stages of membrane protein biogenesis, revealing a dynamic set of steps that imply an unexpectedly flexible membrane insertion machinery.

  12. Cloning and overexpression of antifungal barley chitinase gene in Escherichia coli.

    Science.gov (United States)

    Kirubakaran, S Isaac; Sakthivel, N

    2007-03-01

    Plant chitinases are pathogenesis-related proteins, which are believed to be involved in plant defense responses to pathogen infection. In this study, chitinase gene from barley was cloned and overexpressed in Escherichia coli. Chitinase (35 kDa) was isolated and purified. Since the protein was produced as insoluble inclusion bodies, the protein was solubilized and refolded. Purified chitinase exerted broad-spectrum antifungal activity against Botrytis cinerea (blight of tobacco), Pestalotia theae (leaf spot of tea), Bipolaris oryzae (brown spot of rice), Alternaria sp. (grain discoloration of rice), Curvularia lunata (leaf spot of clover) and Rhizoctonia solani (sheath blight of rice). Due to the potential of broad-spectrum antifungal activity barley chitinase gene can be used to enhance fungal-resistance in crop plants such as rice, tobacco, tea and clover. PMID:17029984

  13. Cerebrospinal fluid chitinase-3-like 2 and chitotriosidase are potential prognostic biomarkers in early multiple sclerosis

    DEFF Research Database (Denmark)

    Møllgaard, M; Vinter, Matilda Degn; Sellebjerg, F;

    2016-01-01

    BACKGROUND AND PURPOSE: The role of chitinases and chitinase-like proteins in multiple sclerosis (MS) is currently unknown; however, cerebrospinal fluid (CSF) levels of chitinase 3-like 1 (CHI3L1) predict prognosis in early MS. Whether this applies to other chitinases and chitinase-like proteins is......) and cognitive impairment by the Paced Auditory Serial Addition Test (P = 0.0357, linear regression) at follow-up. In a multivariate analysis of MS risk, CHI3L2 performed better than CHI3L1. CONCLUSIONS: CHI3L2 and chitotriosidase are promising biomarkers in patients with a first demyelinating episode......, immunoglobulin G index and leukocyte count were investigated. Long-term MS risk and disability (Expanded Disability Status Scale, Multiple Sclerosis Functional Composite components) were examined in a retrospective cohort of 78 patients with ON as the first demyelinating episode (mean follow-up 14 years). The...

  14. Purification, crystallization and preliminary X-ray crystallographic analysis of chitinase from Bacillus cereus NCTU2

    International Nuclear Information System (INIS)

    The crystallization of B. cereus chitinase is reported. Chitinases (EC 3.2.1.14) are found in a broad range of organisms, including bacteria, fungi and higher plants, and play different roles depending on their origin. A chitinase from Bacillus cereus NCTU2 (ChiNCTU2) capable of hydrolyzing chitin as a carbon and nitrogen nutrient has been identified as a member of the family 18 glycoside hydrolases. ChiNCTU2 of molecular weight 36 kDa has been crystallized using the hanging-drop vapour-diffusion method. According to the diffraction of chitinase crystals at 1.10 Å resolution, the crystal belongs to space group P21, with unit-cell parameters a = 50.79, b = 48.79, c = 66.87 Å, β = 99.31°. Preliminary analysis indicates there is one chitinase molecule in the asymmetric unit, with a solvent content of 43.4%

  15. Low chitinase activity in Acacia myrmecophytes: a potential trade-off between biotic and chemical defences?

    Science.gov (United States)

    Heil, M.; Staehelin, Christian; McKey, D.

    We determined chitinase activity in leaves of four myrmecophytic and four non-myrmecophytic leguminous species at the plants' natural growing sites in Mexico. Myrmecophytic plants (or 'ant plants') have obligate mutualisms with ants protecting them against herbivores and pathogenic fungi. Plant chitinases can be considered a reliable measure of plant resistance to pathogenic fungi. The myrmecophytic Acacia species, which were colonised by mutualistic ants, exhibited at least six-fold lower levels of chitinase activity compared with the non-myrmecophytic Acacia farnesiana and three other non-myrmecophytes. Though belonging to different phylogenetic groups, the myrmecophytic Acacia species formed one distinct group in the data set, which was clearly separated from the non-myrmecophytic species. These findings allowed for comparison between two recent hypotheses that attempt to explain low chitinase activity in ant plants. Most probably, chitinases are reduced in myrmecophytic plant species because these are effectively defended indirectly due to their symbiosis with mutualistic ants.

  16. EXPRESSION OF CHITINASE GENE IN TRANSGENIC RAPE PLANTS

    Directory of Open Access Journals (Sweden)

    Lu Longdou

    2005-08-01

    Full Text Available The hypocotyl and cotyledon of Brassica napus L. H165 and Brassica juncea DB3 were transformed with chitinase gene and herbicide-resistance gene by co-culture with Agrobacterium tumefacients LBA4404, and rape plants were obtained which could grow on the medium containing herbicide. The PCR result showed that exotic genes were integrated in the genome of the rape. Further study was performed to determine the impact of temperature on the transgenic rate and the differentiation of explants.

  17. Equilibrium heat-induced denaturation of chitinase 40 from Streptomyces thermoviolaceus.

    Science.gov (United States)

    Pyrpassopoulos, Serapion; Vlassi, Metaxia; Tsortos, Achilleas; Papanikolau, Yannis; Petratos, Kyriacos; Vorgias, Constantinos E; Nounesis, George

    2006-08-01

    High-precision differential scanning calorimetry (DSC) and circular dichroism (CD) have been employed to study the thermal unfolding of chitinase 40 (Chi40) from Streptomyces thermoviolaceus. Chi40 belongs to family 18 of glycosyl hydrolase superfamily bearing a catalytic domain with a "TIM barrel"-like fold, which exhibits deviations from the (beta/alpha)8 fold. The thermal unfolding is reversible at pH = 8.0 and 9.0. The denatured state is characterized by extensive structural changes with respect to the native. The process is characterized by slow relaxation kinetics. Even slower refolding rates are recorded upon cooling. It is shown that the denaturation calorimetric data obtained at slow heating rate (0.17 K/min) are in excellent agreement with equilibrium data obtained by extrapolation of the experimental results to zero scanning rate. Analysis of the DSC results reveals that the experimental data can be successfully fitted using either a non-two-state sequential model involving one equilibrium intermediate, or an independent transitions model involving the unfolding of two Chi40 energetic domains to intermediate states. The stability of the native state with respect to the final denatured state is estimated, deltaG = 24.0 kcal/mol at 25 degrees C. The thermal results are in agreement with previous findings from chemical denaturation studies of a wide variety of (beta/alpha)8 barrel proteins, that their unfolding is a non-two-state process, always involving at least one unfolding intermediate. PMID:16685709

  18. Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Application as Biocontrol for Aphis gossypii

    Directory of Open Access Journals (Sweden)

    TARUNI SRI PRAWASTI

    2010-12-01

    Full Text Available Chitin, a common constituent of insect exoskeleton, could be hydrolyzed by chitinase. This research was conducted to select rhizobacteria isolated from the rhizosphere of chili pepper that produced chitinase and to examine their chitinase activity in degrading chitin of the Aphis gossypii. A total of 25 rhizobacteria isolates formed a clear zone when grown on chitin agar. Three of them had the highest chitinolytic index and were identified as Bacillus sp. strain I.5, I.21, and II.14. The II.14 was chosen for characterization of chitinase activity. The isolate showed maximum chitinase activity at 48-h-incubation. Maximum temperature and pH of the chitinase activity were 55°C and 7.0, respectively. The cell culture and the enzyme crude extract of the above three isolates were tested against A. gossypii and the result was compared to the control through microscopic observation. Hydrolytic analysis showed that the enzyme crude extract of these isolates were able to degrade chitin of insect exoskeleton since the first 3-h-incubation. Meanwhile, the cell culture treatment on the chitin showed degrading activity after 12 h (Bacillus sp. strain I.21 and II.14, and 9 h (Bacillus sp. strain I.5. Chitin degradation of A. gossypii exoskeleton by enzyme crude extract was better than the cell culture treatment. Chitinases produced by Bacillus sp. strains I.5, I.21, and II.14 are potential as biocontrol agents for A. gossypii.

  19. Differential chitinase activity in banana cultivars as a response to Fusarium oxysporum f. sp. cubense infection

    International Nuclear Information System (INIS)

    Six banana clones with varying levels of resistance were inoculated with conidial suspension of races 1 and 4 of Fusarium oxysporum f. sp. cubense (FOC). Chitinase activity in the corm and root tissues was monitored before and after infection to relate with the field resistance or susceptibility of banana cultivars. Resistant clones showed high constitutive chitinase activity in roots and a rapid response to infection. The results suggest that chitinase could be considered as part of a complex mechanism leading to disease resistance. (author)

  20. Chitinase genes revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a phytopathogen suppressive soil

    Energy Technology Data Exchange (ETDEWEB)

    Hjort, K.; Bergstrom, M.; Adesina, M.F.; Jansson, J.K.; Smalla, K.; Sjoling, S.

    2009-09-01

    Soil that is suppressive to disease caused by fungal pathogens is an interesting source to target for novel chitinases that might be contributing towards disease suppression. In this study we screened for chitinase genes, in a phytopathogen-suppressive soil in three ways: (1) from a metagenomic library constructed from microbial cells extracted from soil, (2) from directly extracted DNA and (3) from bacterial isolates with antifungal and chitinase activities. Terminal-restriction fragment length polymorphism (T-RFLP) of chitinase genes revealed differences in amplified chitinase genes from the metagenomic library and the directly extracted DNA, but approximately 40% of the identified chitinase terminal-restriction fragments (TRFs) were found in both sources. All of the chitinase TRFs from the isolates were matched to TRFs in the directly extracted DNA and the metagenomic library. The most abundant chitinase TRF in the soil DNA and the metagenomic library corresponded to the TRF{sup 103} of the isolate, Streptomyces mutomycini and/or Streptomyces clavifer. There were good matches between T-RFLP profiles of chitinase gene fragments obtained from different sources of DNA. However, there were also differences in both the chitinase and the 16S rRNA gene T-RFLP patterns depending on the source of DNA, emphasizing the lack of complete coverage of the gene diversity by any of the approaches used.

  1. Heterologous expression and characterization of two chitinase 5 enzymes from the migratory locust Locusta migratoria.

    Science.gov (United States)

    Li, Ying-Long; Song, Hui-Fang; Zhang, Xue-Yao; Li, Da-Qi; Zhang, Ting-Ting; Ma, En-Bo; Zhang, Jian-Zhen

    2016-06-01

    Insect chitinases are involved in degradation of chitin from the exoskeleton or peritrophic metrix of midgut. In Locusta migratoria, two duplicated Cht5s (LmCht5-1 and LmCht5-2) have been shown to have distinct molecular characteristics and biological roles. To explore the protein properties of the two LmCht5s, we heterologously expressed both enzymes using baculovirus expression system in SF9 cells, and characterized kinetic and carbohydrate-binding properties of purified enzymes. LmCht5-1 and LmCht5-2 exhibited similar pH and temperature optimums. LmCht5-1 has lower Km value for the oligomeric substrate (4MU-(GlcNAc)3 ), and higher Km value for the longer substrate (CM-Chitin-RBV) compared with LmCht5-2. A comparison of amino acids and homology modeling of catalytic domain presented similar TIM barrel structures and differentiated amino acids between two proteins. LmCht5-1 has a chitin-binding domain (CBD) tightly bound to colloidal chitin, but LmCht5-2 does not have a CBD for binding to colloidal chitin. Our results suggested both LmCht5-1 and LmCht5-2, which have the critical glutamate residue in region II of catalytic domain, exhibited chitinolytic activity cleaving both polymeric and oligomeric substrates. LmCht5-1 had relatively higher activity against the oligomeric substrate, 4MU-(GlcNAc)3 , whereas LmCht5-2 exhibited higher activity toward the longer substrate, CM-Chitin-RBV. These findings are helpful for further research to clarify their different roles in insect growth and development. PMID:26792119

  2. Antifungal chitinase against human pathogenic yeasts from Coprinellus congregatus.

    Science.gov (United States)

    Yoo, Yeeun; Choi, Hyoung T

    2014-05-01

    The inky cap, Coprinellus congregatus, produces mushrooms which become autolyzed rapidly to generate black liquid droplets, in which no cell wall is detected by microscopy. A chitinase (Chi2) which is synthesized during the autolytic phase of C. congregatus inhibits the growths of Candida albicans and Cryptococcus neoformans up to 10% at the concentration of 10 μg/ml, about 50% at concentration of 20 μg/ml, and up to 95% at the concentration of 70 μg/ml. Upon treatment these yeast cells are observed to be severely deformed, with the formation of large holes in the cell wall. The two yeast species show no growth inhibition at the concentration of 5 μg/ml, which means the minimum inhibitory concentrations for both yeast species are 10 μg/ml under these experimental conditions. PMID:24535739

  3. Dual protonophore-chitinase inhibitors dramatically affect O. volvulus molting.

    Science.gov (United States)

    Gooyit, Major; Tricoche, Nancy; Lustigman, Sara; Janda, Kim D

    2014-07-10

    The L3-stage-specific chitinase OvCHT1 has been implicated in the development of Onchocerca volvulus, the causative agent of onchocerciasis. Closantel, a known anthelmintic drug, was previously discovered as a potent and specific OvCHT1 inhibitor. As closantel is also a known protonophore, we performed a simple scaffold modulation to map out the structural features that are relevant for its individual or dual biochemical roles. Furthermore, we present that either OvCHT1 inhibition or protonophoric activity was capable of affecting O. volvulus L3 molting and that the presence of both activities in a single molecule yielded more potent inhibition of the nematode's developmental process. PMID:24918716

  4. Purification and characterization of thermostable chitinase from a novel S. maltophilia strain

    Directory of Open Access Journals (Sweden)

    Javed, S.

    2013-01-01

    Full Text Available Aims: The presents study examines the purification and characterization of a chitinase from S. maltophilia SJ602 strainisolated from a soil sample collected from Jamia Hamdard, New Delhi.Methodology and Results: The purification steps included chitin affinity using colloidal chitin as the affinity matrix andcolumn chromatography using Sephadex G-100. The chitinase was purified to 66 fold having a yield of 17%. The molecular weight of the chitinase was found to be around 29 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE. The pH and temperature optima of the purified chitinase were found to be at pH 5.5 and60 °C, respectively. Conclusion, Significance and Impact of the study: Besides showing a significant yield, the enzyme has a highthermal stability which has its applicability in the recycling of chitin waste.

  5. Production of chitinases with Trichoderma harzianun isolates using solid substrate fermentation

    Institute of Scientific and Technical Information of China (English)

    Viviana Nagy

    2004-01-01

    @@ Over forty Trichoderma harzianum isolates have been screened in solid substrate fermentation (SSF)for chitinase production. Strains were isolated from Asian soil and tree bark samples. Identification was performed in Canada and Austria by classical and molecular taxonomical methods.

  6. Chitinase-mediated inhibitory activity of Brassica transgenic on growth of Alternaria brassicae.

    Science.gov (United States)

    Mondal, Kalyan K; Chatterjee, Subhas Chandra; Viswakarma, Navin; Bhattacharya, Ram Charan; Grover, Anita

    2003-09-01

    Chitinase, capable of degrading the cell walls of invading phytopathogenic fungi, plays an important role in plant defense response, particularly when this enzyme is overexpressed through genetic engineering. In the present study, Brassica plant (Brassica juncea L.) was transformed with chitinase gene tagged with an overexpressing promoter 35 S CaMV. The putative transgenics were assayed for their inhibitory activity against Alternaria brassicae, the inducer of Alternaria leaf spot of Brassica both in vitro and under polyhouse conditions. In in vitro fungal growth inhibition assays, chitinase inhibited the fungal colony size by 12-56% over the non-trangenic control. The bioassay under artificial epiphytotic conditions revealed the delay in the onset of disease as well as reduced lesion number and size in 35S-chitinase Brassica as compared to the untransformed control plants. PMID:14570264

  7. Isolation, partial characterization, and cloning of an extracellular chitinase from the entomopathogenic fungus Verticillium lecanii.

    Science.gov (United States)

    Yu, G; Xie, L Q; Li, J T; Sun, X H; Zhang, H; Du, Q; Li, Q Y; Zhang, S H; Pan, H Y

    2015-01-01

    The entomopathogenic fungus Verticillium lecanii is a well-known biocontrol agent of fungal phytopathogens, as well as insect pests. A 42-kDa chitinase belonging to family 18 of the glycosyl hydrolases was isolated and partially characterized. Chitinase was purified using successive column chromatography on phenyl-sepharose, DEAE-sepharose, and CM-sepharose. The enzyme showed the highest activity at 40°C and pH 4.6. Enzyme activity was strongly activated in the presence of Mg(2+). The purified enzyme showed inhibitory activity of spore germination against several plant pathogens, particularly Fusarium moniliforme. The genomic DNA and cDNA sequences were resolved by polymerase chain reaction amplification and sequencing. Protein modeling and comparative investigation of different chitinase amino acids showed that chitinases are conserved in parasitic fungi. PMID:25867374

  8. Aromatic-Mediated Carbohydrate Recognition in Processive Serratia marcescens Chitinases.

    Science.gov (United States)

    Jana, Suvamay; Hamre, Anne Grethe; Wildberger, Patricia; Holen, Matilde Mengkrog; Eijsink, Vincent G H; Beckham, Gregg T; Sørlie, Morten; Payne, Christina M

    2016-02-25

    Microorganisms use a host of enzymes, including processive glycoside hydrolases, to deconstruct recalcitrant polysaccharides to sugars. Processive glycoside hydrolases closely associate with polymer chains and repeatedly cleave glycosidic linkages without dissociating from the crystalline surface after each hydrolytic step; they are typically the most abundant enzymes in both natural secretomes and industrial cocktails by virtue of their significant hydrolytic potential. The ubiquity of aromatic residues lining the enzyme catalytic tunnels and clefts is a notable feature of processive glycoside hydrolases. We hypothesized that these aromatic residues have uniquely defined roles, such as substrate chain acquisition and binding in the catalytic tunnel, that are defined by their local environment and position relative to the substrate and the catalytic center. Here, we investigated this hypothesis with variants of Serratia marcescens family 18 processive chitinases ChiA and ChiB. We applied molecular simulation and free energy calculations to assess active site dynamics and ligand binding free energies. Isothermal titration calorimetry provided further insight into enthalpic and entropic contributions to ligand binding free energy. Thus, the roles of six aromatic residues, Trp-167, Trp-275, and Phe-396 in ChiA, and Trp-97, Trp-220, and Phe-190 in ChiB, have been examined. We observed that point mutation of the tryptophan residues to alanine results in unfavorable changes in the free energy of binding relative to wild-type. The most drastic effects were observed for residues positioned at the "entrances" of the deep substrate-binding clefts and known to be important for processivity. Interestingly, phenylalanine mutations in ChiA and ChiB had little to no effect on chito-oligomer binding, in accordance with the limited effects of their removal on chitinase functionality. PMID:26824449

  9. ISOLATION AND CHARACTERIZATION OF CHITINASE GENE FROM THE UNTRADITIONAL PLANT SPECIES

    OpenAIRE

    Dominika Ďurechová; Ildikó Matušíková; Jana Moravčíková; Martin Jopčík; Jana Libantová

    2013-01-01

    Round-leaf sundew (Drosera rotundifolia L.) from Droseraceae family belongs among a few plant species with strong antifungal potential. It was previously shown that chitinases of carnivorous plant species may play role during the insect prey digestion, when hard chitin skeleton is being decomposed. As many phytopathogenic fungi contain chitin in their cell wall our attention in this work was focused on isolation and in silico characterization of genomic DNA sequence of sundew chitinase gene. ...

  10. Draft Genome Sequence of a Chitinase-producing Biocontrol Bacterium Serratia sp. C-1

    OpenAIRE

    Seur Kee Park; Young Cheol Kim

    2015-01-01

    The chitinase-producing bacterial strain C-1 is one of the key chitinase-producing biocontrol agents used for effective bioformulations for biological control. These bioformulations are mixed cultures of various chitinolytic bacteria. However, the precise identification, biocontrol activity, and the underlying mechanisms of the strain C-1 have not been investigated so far. Therefore, we evaluated in planta biocontrol efficacies of C-1 and determined the draft genome sequence of the strain in ...

  11. Fungal chitinases: function, regulation, and potential roles in plant/pathogen interactions.

    Science.gov (United States)

    Langner, Thorsten; Göhre, Vera

    2016-05-01

    In the past decades our knowledge about fungal cell wall architecture increased tremendously and led to the identification of many enzymes involved in polysaccharide synthesis and remodeling, which are also of biotechnological interest. Fungal cell walls play an important role in conferring mechanic stability during cell division and polar growth. Additionally, in phytopathogenic fungi the cell wall is the first structure that gets into intimate contact with the host plant. A major constituent of fungal cell walls is chitin, a homopolymer of N-acetylglucosamine units. To ensure plasticity, polymeric chitin needs continuous remodeling which is maintained by chitinolytic enzymes, including lytic polysaccharide monooxygenases N-acetylglucosaminidases, and chitinases. Depending on the species and lifestyle of fungi, there is great variation in the number of encoded chitinases and their function. Chitinases can have housekeeping function in plasticizing the cell wall or can act more specifically during cell separation, nutritional chitin acquisition, or competitive interaction with other fungi. Although chitinase research made huge progress in the last decades, our knowledge about their role in phytopathogenic fungi is still scarce. Recent findings in the dimorphic basidiomycete Ustilago maydis show that chitinases play different physiological functions throughout the life cycle and raise questions about their role during plant-fungus interactions. In this work we summarize these functions, mechanisms of chitinase regulation and their putative role during pathogen/host interactions. PMID:26527115

  12. High-capacity calcium-binding chitinase III from pomegranate seeds (Punica granatum Linn.) is located in amyloplasts

    OpenAIRE

    Lv, Chenyan; Masuda, Taro; Yang, Haixia; Sun, Lei; Zhao, Guanghua

    2011-01-01

    We have recently identified a new class III chitinase from pomegranate seeds (PSC). Interestingly, this new chitinase naturally binds calcium ions with high capacity and low affinity, suggesting that PSC is a Ca-storage protein. Analysis of the amino acid sequence showed that this enzyme is rich in acidic amino acid residues, especially Asp, which are responsible for calcium binding. Different from other known chitinases, PSC is located in the stroma of amyloplasts in pomegranate seeds. Trans...

  13. Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A

    Science.gov (United States)

    Zarei, Mandana; Aminzadeh, Saeed; Zolgharnein, Hossein; Safahieh, Alireza; Daliri, Morteza; Noghabi, Kambiz Akbari; Ghoroghi, Ahmad; Motallebi, Abbasali

    2011-01-01

    Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. In the following investigation, a novel chitinase with antifungal activity was characterized from a native Serratia marcescens B4A. Partially purified enzyme had an apparent molecular mass of 54 kDa. It indicated an optimum activity in pH 5 at 45°C. Enzyme was stable in 55°C for 20 min and at a pH range of 3–9 for 90 min at 25°C. When the temperature was raised to 60°C, it might affect the structure of enzymes lead to reduction of chitinase activity. Moreover, the Km and Vmax values for chitin were 8.3 mg/ml and 2.4 mmol/min, respectively. Additionally, the effect of some cations and chemical compounds were found to stimulate the chitinase activity. In addition, Iodoacetamide and Idoacetic acid did not inhibit enzyme activity, indicating that cysteine residues are not part of the catalytic site of chitinase. Finally, chitinase activity was further monitored by scanning electronic microscopy data in which progressive changes in chitin porosity appeared upon treatment with chitinase. This enzyme exhibited antifungal activity against Rhizoctonia solani, Bipolaris sp, Alternaria raphani, Alternaria brassicicola, revealing a potential application for the industry with potentially exploitable significance. Fungal chitin shows some special features, in particular with respect to chemical structure. Difference in chitinolytic ability must result from the subsite structure in the enzyme binding cleft. This implies that why the enzyme didn’t have significant antifungal activity against other Fungi. PMID:24031719

  14. Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A

    Directory of Open Access Journals (Sweden)

    Mandana Zarei

    2011-09-01

    Full Text Available Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. In the following investigation, a novel chitinase with antifungal activity was characterized from a native Serratia marcescens B4A. Partially purified enzyme had an apparent molecular mass of 54 kDa. It indicated an optimum activity in pH 5 at 45ºC. Enzyme was stable in 55ºC for 20 min and at a pH range of 3-9 for 90 min at 25ºC. When the temperature was raised to 60ºC, it might affect the structure of enzymes lead to reduction of chitinase activity. Moreover, the Km and Vmax values for chitin were 8.3 mg/ml and 2.4 mmol/min, respectively. Additionally, the effect of some cations and chemical compounds were found to stimulate the chitinase activity. In addition, Iodoacetamide and Idoacetic acid did not inhibit enzyme activity, indicating that cysteine residues are not part of the catalytic site of chitinase. Finally, chitinase activity was further monitored by scanning electronic microscopy data in which progressive changes in chitin porosity appeared upon treatment with chitinase. This enzyme exhibited antifungal activity against Rhizoctonia solani, Bipolaris sp, Alternaria raphani, Alternaria brassicicola, revealing a potential application for the industry with potentially exploitable significance. Fungal chitin shows some special features, in particular with respect to chemical structure. Difference in chitinolytic ability must result from the subsite structure in the enzyme binding cleft. This implies that why the enzyme didn't have significant antifungal activity against other Fungi.

  15. Isolation and purification of fungal pathogen (Macrophomina phaseolina induced chitinase from moth beans (Phaseolus aconitifolius

    Directory of Open Access Journals (Sweden)

    Neelima Garg

    2010-01-01

    Full Text Available Objective : Chitinase (EC 3.2.1.14 is one of the major pathogenesis-related proteins, which is a polypeptide that accumulates extracellularly in infected plant tissue. An attempt was made to isolate and purify the chitanase enzyme using moth beans as an enzyme source. Materials and Method : The enzyme was isolated and purified from moth beans against the fungal pathogen Macrophomina phaseolina strain 2165. The isolation and purification was done in both in vitro and in vivo conditions. Purification of chitinase was carried out to obtain three fractions, viz. 50°C heated, ammonium sulfate precipitated and sephadex G-25 column-eluted fractions. The molecular mass of Chitinase was directly estimated by sodium dodecyl sulfate-polyacryamide gel electroresis (SDS-PAGE. Result : The yield is sufficient for initial characterization studies of the enzyme. The molecular study of the enzyme shows the possibility of generating the defense mechanism in plants in which it cannot occur. Chitinase was purified by gel filtration chromatography with 20.75-fold and 32.78-fold purification in the in vitro and in vivo conditions, respectively. The enzyme shows a maximum activity after 90 min with 0.1 ml of colloidal chitin as a substrate and 0.4 ml of crude chitinase extract. The optimum pH of 5.0 and an optimum temperature of 40°C was found for maximal activity. The molecular weight of purified chitinase was estimated to be 30 kDa by SDS-PAGE. Conclusion : The chitinase isolated in both in vitro and in vivo conditions is stable andactive.

  16. Purification and characterization of chitinase from Bacillus circulans No.4.1.

    Science.gov (United States)

    Wiwat, C; Siwayaprahm, P; Bhumiratana, A

    1999-09-01

    Bacillus circulans No.4.1 produced a high level of chitinase when cells were grown in tryptic soy broth supplemented with 0.3% colloidal chitin at 35 degrees C for 5 days. Purification was carried out by protein precipitation with 80% saturation ammonium sulfate, anion-exchange chromatography with DEAE-Sephacel, and gel filtration with Sephadex G-100, sequentially. The purified enzyme could be demonstrated as a single band on SDS-PAGE, estimated to be 45 kDa. This enzyme could hydrolyze colloidal chitin, purified chitin, glycol chitin, carboxymethyl-chitin (CM-chitin), and 4-methylumbelliferyl-beta-D-N,N'-diacetylchitobioside [4-MU-(GlcNAc)(2)]. The optimal conditions for this chitinase were pH 8.0 and 40 degrees C. The isoelectric point of the chitinase was 5.1. The amino acid composition of the purified chitinase was determined. The initial 20 amino acid residues of the N-terminal were found to be alanine (A), proline (P), tryptophan (W), asparagine (N), serine (S), lysine (K), glycine (G), asparagine (N), tyrosine (Y), alanine (A), leucine (L), proline (P), tyrosine (Y), tyrosine (Y), arginine (R), glycine (G), alanine (A), tryptophan (W), alanine (A), and valine (V). Knowledge of these properties of chitinase from B. circulans No. 4.1 should be useful in the development of genetically engineered Bacillus sp. as biopesticides. PMID:10441726

  17. An investigation of a defensive chitinase against Fusarium oxysporum in pepper leaf tissue

    Directory of Open Access Journals (Sweden)

    Khemika S. Lomthaisong

    2008-01-01

    Full Text Available Plant chitinase is classified as a PR-protein involved in a defense mechanism against a pathogen. This research aims to investigate a specific type of chitinase which is produced by pepper in response to an early defense against Fusarium oxysporum, which causes wilt disease. The changes of chitinase isozyme patterns in the inter- and intracellular fluids in the leaf of four cultivars of pepper (Capsicum annuum L. at day 1, 3, 5, 7 and 10 from fungal inoculation were analysed using SDS-PAGE in polyacrylamide gel supplemented with glycol chitin as a substrate. The levels of disease severity in the four varieties of pepper were also compared with the isozyme patterns. The results showed that the resistance of pepper to F. oxysporum attack corresponded to the expression of ~70 kDa chitinase band (Chi-3 in the intercellular fluid. Therefore, such chitinase could possibly be used as a protein marker to identify the tolerant line and as a springboard for further study of wilt disease control.

  18. Characterization of two Listeria innocua chitinases of different sizes that were expressed in Escherichia coli.

    Science.gov (United States)

    Honda, Shotaro; Wakita, Satoshi; Sugahara, Yasusato; Kawakita, Masao; Oyama, Fumitaka; Sakaguchi, Masayoshi

    2016-09-01

    Two putative chitinase genes, lin0153 and lin1996, from the nonpathogenic bacterium Listeria innocua were expressed in Escherichia coli, and the gene products were characterized. The genes were close homologs of chitinases from the pathogenic bacterium Listeria monocytogenes, in which chitinases and chitin-binding proteins play important roles in pathogenesis in mice-infection models. The purified recombinant enzymes that are different in size, LinChi78 (lin0153 product) and LinChi35 (lin1996 product)-with molecular masses of 82 and 38 kDa, including vector-derived additional sequences, respectively-exhibited optimum catalytic activity under neutral and acidic conditions at 50 °C, respectively, and were stable over broad pH (4-11) and temperature (4-40 °C) ranges. LinChi35 displayed higher k cat and K M values for 4-nitrophenyl N,N-diacetyl-β-D-chitobioside [4NP-(GlcNAc)2] than LinChi78. Both enzymes produced primarily dimers from colloidal chitin as a substrate. However, LinChi78 and LinChi35 could hydrolyze oligomeric substrates in a processive exo- and nonprocessive endo-manner, respectively, and showed different reactivity toward oligomeric substrates. Both enzymes could bind chitin beads but were different in their binding ability toward crystalline α-chitin and cellulose. The structure-function relationships of these chitinases are discussed in reference to other bacterial chitinases. PMID:27138200

  19. [Suprapubic catheter insertion].

    Science.gov (United States)

    Neumann, Eva; Schwentner, Christian

    2016-01-01

    The suprapubic catheter enables a percutaneous drainage of urine. The insertion is made superior of the pubic bone through the abdominal wall into the bladder. It allows a permanent drainage of urine bypassing the urethra. The insertion of a suprapubic catheter requires knowledge and expertise. This paper summarizes the basic background and allows to follow the practical application step by step. PMID:26800072

  20. Tie rod insertion test

    CERN Multimedia

    B. LEVESY

    2002-01-01

    The superconducting coil is inserted in the outer vaccum tank and supported by a set of tie rods. These tie rods are made of titanium alloy. This test reproduce the final insertion of the tie rods inside the outer vacuum tank.

  1. Degradation of chitin and chitosan by a recombinant chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    Science.gov (United States)

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  2. Comparative molecular evolution of Trichoderma chitinases in response to mycoparasitic interactions

    DEFF Research Database (Denmark)

    Ihrmark, Katarina; Asmail, Nashwan; Ubhayasekera, Wimal;

    2010-01-01

    Certain species of the fungal genus Trichoderma are potent mycoparasites and are used for biological control of fungal diseases on agricultural crops. In Trichoderma, whole-genome sequencing reveal between 20 and 36 different genes encoding chitinases, hydrolytic enzymes that are involved in the...... mycoparasitic attack. Sequences of Trichoderma chitinase genes chi18-5, chi18-13, chi18-15 and chi18-17, which all exhibit specific expression during mycoparasitism-related conditions, were determined from up to 13 different taxa and studied with regard to their evolutionary patterns. Two of them, chi18-13 and...... Trichoderma clades are observed. These observations show that Trichoderma chitinases chi18-13 and chi18-15 evolve in a manner consistent with rapid co-evolutionary interactions and identifies putative target regions involved in determining substrate-specificity....

  3. Are mycoparasitism and chitinase production species or isolate dependent in Trichoderma ?

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Nagy V; Kovacs K

    2004-01-01

    @@ The relationship between taxonomic status of Trichoderma spp., chitinase production in solid substrate fermentation (SSF) on four media and mycoparasitism in dual culture (confrontation assay)against four plant pathogenic fungi was studied. Seventy five Trichoderma isolates belonging to 35species have been screened. The plant pathogenic fungi used in confrontation assay were Botrytis cinerea , Fusarium oxysporum f. sp. dianthi , Rhizoctonia solani and Sclerotinia sclerotiorum . The SSF media contained wheat bran, crude chitin (from crab shells, SIGMA) and salt solutions. The best performing isolates in mycoparasitism tests were Trichoderma flavofuscum, T. harzianum, T.inhamatum, T. koningii and T. strigosum. Some isolates exhibiting good mycoparasitism produced chitinase in SSF only at low or medium level. In contrary there were isolates with excellent extracellular chitinase production but their biocontrol potential did not belong to the leading group.Statistical methods have been used to evaluate the data.

  4. Significance of Penicillium ochrochloron chitinase as a biocontrol agent against pest Helicoverpa armigera.

    Science.gov (United States)

    Patil, Nilambari S; Jadhav, Jyoti P

    2015-06-01

    Penicillium ochrochloron chitinase purified by DEAE-cellulose ion exchange chromatography was evaluated for its antifeedant and growth inhibitory activities against Helicoverpa armigera at different concentrations of 2000, 1000, 500, 250 and 100 U mL(-1). It reduced the successful pupation and increased larval and pupal mortality, adult emergence in a dosage-dependent manner when applied topically. The highest mortalities were recorded for groups treated with 2000 U mL(-1) chitinase activity. The studies showed P.ochrochloron chitinase can affect the growth of H.armigera larvae. Since this insect pest species has developed resistance and resurgence to chemical insecticides, only alternate is the usage of enzyme-based pesticide formulations as an environmentally friendly pest management tool. PMID:25723715

  5. Extraction and partial purification of chitinase from mycosymbiont and its relation with mycorrhiza association process

    International Nuclear Information System (INIS)

    Extraction effectivity and partial purification of chitinase extracellular metabolite from Scleroderma columnare, Pisolithus tinctorius, Trichoderma harzianum and the root of Shorea selanica have been searched. S. columnare and P. tinctorius were the mycosymbiont for S. selanica while T. harzianum was not. (NH4)2SO4 was the very effective solvent for crude extracellular enzyme extraction, followed by PEG-6000 and ethanol 50 percent respectively. The activity of P. tinctorius was the lowest among the microbe while S. columnare was the highest. The activity lowered by purification process but specific activity was not. The chitinase activity of inoculated root was higher in the S. columnare association than P. tinctorius's also the percentage of root chitin content and infection rate. It mean that S. columnare more effective as mycosymbiont. The periods of association lowered the activity of root chitinase but this phenomenon did not happen in the root of S. selanica with T. harzianum infection

  6. Insectivorous bats digest chitin in the stomach using acidic mammalian chitinase.

    Science.gov (United States)

    Strobel, Sara; Roswag, Anna; Becker, Nina I; Trenczek, Tina E; Encarnação, Jorge A

    2013-01-01

    The gastrointestinal tract of animals is adapted to their primary source of food to optimize resource use and energy intake. Temperate bat species mainly feed on arthropods. These contain the energy-rich carbohydrate chitin, which is indigestible for the endogenous enzymes of a typical mammalian gastrointestinal tract. However, the gastrointestinal tract of bat species should be adapted to their diet and be able to digest chitin. We hypothesized that (i) European vespertilionid bat species have the digestive enzyme chitinase and that (ii) the chitinolytic activity is located in the intestine, as has been found for North American bat species. The gastrointestinal tracts of seven bat species (Pipistrellus pipistrellus, Plecotus auritus, Myotis bechsteinii, Myotis nattereri, Myotis daubentonii, Myotis myotis, and Nyctalus leisleri) were tested for chitinolytic activity by diffusion assay. Gastrointestinal tracts of P. pipistrellus, P. auritus, M. nattereri, M. myotis, and N. leisleri were examined for acidic mammalian chitinase by western blot analysis. Tissue sections of the gastrointestinal tract of P. pipistrellus were immunohistochemically analyzed to locate the acidic mammalian chitinase. Chitinolytic activity was detected in the stomachs of all bat species. Western blot analysis confirmed the acidic mammalian chitinase in stomach samples. Immunohistochemistry of the P. pipistrellus gastrointestinal tract indicated that acidic mammalian chitinase is located in the stomach chief cells at the base of the gastric glands. In conclusion, European vespertilionid bat species have acidic mammalian chitinase that is produced in the gastric glands of the stomach. Therefore, the gastrointestinal tracts of insectivorous bat species evolved an enzymatic adaptation to their diet. PMID:24019876

  7. Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A

    OpenAIRE

    Mandana Zarei; Saeed Aminzadeh; Hossein Zolgharnein; Alireza Safahieh; Morteza Daliri; Kambiz Akbari Noghabi; Ahmad Ghoroghi; Abbasali Motallebi

    2011-01-01

    Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. In the following investigation, a novel chitinase with antifungal activity was characterized from a native Serratia marcescens B4A. Partially purified enzyme had an apparent molecular mass of 54 kDa. It indicated an optimum activity in pH 5 at 45ºC. Enzyme was stable in 55ºC for 20 min and at a pH range of 3-9 for 90 min at 25ºC. When the temperature w...

  8. Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A

    OpenAIRE

    Zarei, Mandana; Aminzadeh, Saeed; Zolgharnein, Hossein; Safahieh, Alireza; Daliri, Morteza; Noghabi, Kambiz Akbari; Ghoroghi, Ahmad; Motallebi, Abbasali

    2011-01-01

    Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. In the following investigation, a novel chitinase with antifungal activity was characterized from a native Serratia marcescens B4A. Partially purified enzyme had an apparent molecular mass of 54 kDa. It indicated an optimum activity in pH 5 at 45°C. Enzyme was stable in 55°C for 20 min and at a pH range of 3–9 for 90 min at 25°C. When the temperature w...

  9. Study on the character of chitinase produced by Trichoderna spp.with measuring reducing sugar

    Institute of Scientific and Technical Information of China (English)

    LIU Kai-qi; XIANG Mei-mei; LIU Ren; ZENG Yong-san; LI Hua; JIANG Xin-yin; ZHANG Yue-li

    2004-01-01

    @@ A trusty and intuitionistic method for screening chitinase produced by Trichoderma spp. was developed. 38 isolates of Trichoderma spp. were cultured in liquid medium with chitin or colloidal chitin as the sole carbon source for 4 days. The supernatant of the fermented broth was mixed with colloidal chitin and heated in water-bath at 37℃ for 30 min, then 3,5-dinitrosalicylic acid reagent (DNS) was added to the mixture, and let them react for 10 min in water-bath. According to the different colour of the mixture, the isolates of Trichoderma spp. which can produce chitinase could be screened.

  10. Differential response of banana cultivars to F. oxysporum f. sp. cubense infection for Chitinase activity

    International Nuclear Information System (INIS)

    Six banana clones with varying levels of resistance were inoculated with conidial suspension of races 1 and 4 of Fusarium oxysporum f. sp. cubense. Chitanase activity in the corm and root tissues was monitored before and after infection to relate with the field resistance or susceptibility of banana cultivars. Resistant clones showed high constitutive chitinase activity in roots and a rapid response to infection. The results suggest that chitinase could be considered as part of a complex mechanism leading to disease resistance. (author). 5 refs, 8 figs

  11. Acidic mammalian chitinase and the eye: implications for ocular inflammatory diseases

    Directory of Open Access Journals (Sweden)

    ClaudioBucolo

    2011-07-01

    Full Text Available Chitinases have an important role in the defence of organisms against chitin containing parasites. An acidic mammalian chitinase (AMCase has been detected in epithelial cells in lung tissue samples taken from patients with asthma as well as in conjunctival epithelium of patients with inflammatory ocular diseases. Particularly, elevated AMCase activity has been observed in ocular tissues of patients with vernal keratoconjunctivitis, seasonal allergic conjunctivitis, and in patients affected by dry eye syndrome. This enzyme is induced via a TH2-specific, IL-13-dependent pathway. AMCase may thus be a key mediator of IL-13-induced responses in TH2-driven inflammatory ocular diseases.

  12. Antifungal activity of chitinase obtained from Paenibacillus ehimensis MA2012 against conidial of Collectotrichum gloeosporioides in vitro.

    Science.gov (United States)

    Seo, Dong-Jun; Lee, Yong-Sung; Kim, Kil-Yong; Jung, Woo-Jin

    2016-07-01

    To investigate the expression patterns of chitinase on SDS-PAGE gel, Paenibacillus ehimensis MA2012 was incubated in gelatin-chitin medium (GCM) at 30 °C for 7 days. Six major bands (Ch3, Ch4, Ch5, Ch6, Ch7, and Ch8) of chitinase isozymes in GC medium appeared on SDS-PAGE gel during the incubation period. Chitinase activity staining of P. ehimensis MA2012 was detected on 2-DE with different pI values (4-11). After DEAE-Sephadex chromatography, eight bands (Ch1 to Ch8) of chitinase isozymes were stained strongly with Calcofluor white M2R at fraction 45. After Sephadex G-75 gel filtration, six bands (Ch3 to Ch8) of chitinase isozymes were stained with Calcofluor white M2R at fractions of 11-12. The specific activity of the purified chitinase was 3.8 units mg(-1) protein with a purification factor of 0.27. Inhibition rate of the conidial germination of Colletotrichum gloeosporioides was 87% in partial purified chitinase treatment compared with control. PMID:27133265

  13. IN SILICO ANALYSIS OF CHITINASE PROMOTER ISOLATED FROM DROSERA ROTUNDIFOLIA L.

    Directory of Open Access Journals (Sweden)

    Dominika Ďurechová

    2014-02-01

    Full Text Available Chitinases occur in dozens of genes in the individual plant species and play diverse roles in plant growth and development, and during the plant defense to biotic and abiotic stress. Here we focused on isolation and in silico characterization of regulatory sequences of chitinase gene that belongs to the first isolated gene sequences from D. rotundifolia overall. For the isolation of the 739 bp sequence of chitinase promoter the genome walking approach was applied. The authenticity of the obtained fragment(s was verified by sequencing and sequence alignment ClustalW program. The core of the chitinase promoter was predicted by Neural Network Promoter Prediction program. In total the PLACE online available database identified 66 various cis-regulatory elements in the analyzed sequence. Some of them might be potentially bound by specific transcription factors, and regulate gene expression in specific plant tissues during the plant development or upon the pathogen attack, dehydration, cold or high salinity stress. However, further analyses are needed to reveal which out of predicted cis-elements participate in the true expression profile of isolated promoter in origin and transgenic plant organism.

  14. Differential enzymatic activity of common haplotypic versions of the human acidic mammalian chitinase protein

    NARCIS (Netherlands)

    M.A. Seibold; T.A. Reese; S. Choudhry; M.T. Salam; K. Beckman; C. Eng; A. Atakilit; K. Meade; M. Lenoir; H.G. Watson; S. Thyne; R. Kumar; K.B. Weiss; L.C. Grammer; P. Avila; R.P. Schleimer; J.V. Fahy; J. Rodriguez-Santana; W. Rodriguez-Cintron; R.G. Boot; D. Sheppard; F.D. Gilliland; R.M. Locksley; E.G. Burchard

    2009-01-01

    Mouse models have shown the importance of acidic mammalian chitinase activity in settings of Th2 inflammation. AMCase enzymatic activity was necessary for most of the inflammation present in an asthma mouse model. However, in settings of chitin exposure, AMCase-mediated cleavage of the inflammatory

  15. Studies on Exo-Chitinase Production from Trichoderma asperellum UTP-16 and Its Characterization.

    Science.gov (United States)

    Kumar, D Praveen; Singh, Rajesh Kumar; Anupama, P D; Solanki, Manoj Kumar; Kumar, Sudheer; Srivastava, Alok K; Singhal, Pradeep K; Arora, Dilip K

    2012-09-01

    The growth conditions for chitinase production by Trichoderma asperellum UTP-16 in solid state fermentation was optimized using response surface methodology based on central composite design. The chitinase production was optimized, using one-factor at a time approach, with six independent variables (temperature, pH, NaCl, incubation period, nitrogen and carbon sources) and 3.31 Units per gram dry substrate (U gds(-1)) exo-chitinase yield was obtained. A 21.15% increase was recorded in chitinase activity (4.01 U gds(-1)) through surface response methodology, indicates that it is a powerful and rapid tool for optimization of physical and nutritional variables. Further, efficiency of crude enzyme was evaluated against phytopathogenic Fusarium spp. and a mycelial growth inhibition up to 3.5-6.5 mm was achieved in well diffusion assay. These results could be supplemented as basic information for the development of enzyme based formulation of T. asperellum UTP-16 and its use as a biocontrol agent. PMID:23997329

  16. Biological activity of Bacillus thuringiensis (Bacillales: Bacillaceae) chitinase against Caenorhabditis elegans (Rhabditida: Rhabditidae)

    Czech Academy of Sciences Publication Activity Database

    Zhang, L.; Yu, J.; Xie, Y.; Lin, H.; Huang, Z.; Xu, L.; Gelbič, Ivan; Guan, X.

    2014-01-01

    Roč. 107, č. 2 (2014), s. 551-558. ISSN 0022-0493 Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis * Caenorhabditis elegans * chitinase Subject RIV: GF - Plant Pathology, Vermin, Weed, Plant Protection Impact factor: 1.506, year: 2014 http://www.bioone.org/doi/pdf/10.1603/EC13201

  17. Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Its Application as Biocontrol for Whitefly (Bemisia tabaci Genn.

    Directory of Open Access Journals (Sweden)

    Nisa R. Mubarik

    2010-01-01

    Full Text Available Problem statement: Chitin, a common constituent of insect exoskeleton, could be hydrolyzed by chitinase. The research was conducted to screen chitinolytic rhizobacteria isolated from rhizosphere of chilli pepper and to determine their chitinase activity in degrading chitin of whitefly, Bemisia tabaci Genn. (Hemiptera: Aleyrodidae. Whitefly is recognized as an important pest on many crops including chilli pepper. Approach: Screening and molecular identification based on 16S rRNA sequence of chitinolytic isolates, chitinase productions, measurement of chitinase activity, characterization of chitinase and effect of the chitinase treatment on whitefly were studied. Results: A total of 25 isolates of rhizobacteria formed a clear zone on solid chitin media. Two isolates, i.e., I.5 and I.21 isolates had the highest chitinolytic index. Based on sequence of 16S rRNA gene, the isolates of I.5 and I.21 were identified as Bacillus sp. and Bacillus cereus, respectively. The highest chitinolytic index and specific activity of I.5 isolate was 0.94 and 0.11 U mg-1 proteins, respectively. Maximum production of I.5 chitinase was occured after 36 h cultivation at 30°C and pH 7.0. The highest chitinolytic index and specific activity of I.21isolate was 0.75 and 0.114 U mg-1 proteins, respectively. Maximum production of I.21 chitinase was occured after 36 h cultivation at 55°C and pH 7.0. Cell culture and crude enzyme of the isolates were tested on chitin of B. tabaci and the effect was observed using a microscope and sterile water was used as a negative control. Hydrolytic observation showed that crude enzyme of I.21 isolate could degrade chitin of B. tabaci exoskeleton and the activity was better than that of I.5 isolate. Conclusion: Chitinase produced by Bacillus cereus I.21 strain has potential application as biocontrol agents for B. tabaci.

  18. ALS insertion devices

    International Nuclear Information System (INIS)

    The Advanced Light Source (ALS), the first US third generation synchrotron radiation source, is currently under construction at the Lawrence Berkeley Laboratory. The low-emittance, 1.5 GeV electron storage ring and the insertion devices are specifically designed to produce high brightness beams in the UV to soft X-Ray range. The planned initial complement of insertion devices includes four 4.6 m long undulators, with period lengths of 3.9 cm, 5.0 cm (2) and 8.0 cm, and a 2.9 m long wiggler of 16 cm period length. Undulator design is well advanced and fabrication has begun on the 5.0 cm and 8.0 cm period length undulators. This paper discusses ALS insertion device requirements; general design philosophy; and design of the magnetic structure, support structure/drive systems, control system and vacuum system. 18 refs., 9 figs., 5 tabs

  19. Purification and characterization of chitinase from Alcaligenes faecalis AU02 by utilizing marine wastes and its antioxidant activity

    OpenAIRE

    Annamalai, Neelamegam; Veeramuthu Rajeswari, Mayavan; Vijayalakshmi, Shanmugam; Balasubramanian, Thangavel

    2011-01-01

    Marine waste is an abundant renewable source for the recovery of several value added metabolites with potential industrial applications. This study describes the production of chitinase on marine waste, with the subsequent use of the same marine waste for the extraction of antioxidants. A chitinase-producing bacterium isolated from seafood effluent was identified as Alcaligenes faecalis AU02. Optimal chitinase production was obtained in culture conditions of 37°C for 72 h in 100 ml medium con...

  20. Pixel detector insertion

    CERN Multimedia

    CMS

    2015-01-01

    Insertion of the Pixel Tracker, the 66-million-channel device used to pinpoint the vertex of each colliding proton pair, located at the heart of the detector. The geometry of CMS is a cylinder lying on its side (22 meters long and 15 meters high in dia

  1. The Composite Insertion Electrode

    DEFF Research Database (Denmark)

    Atlung, Sven; Zachau-Christiansen, Birgit; West, Keld;

    1984-01-01

    The specific energy obtainable by discharge of porous insertion electrodes is limited by electrolyte depletion in thepores. This can be overcome using a solid ion conductor as electrolyte. The term "composite" is used to distinguishthese electrodes from porous electrodes with liquid electrolyte...

  2. Inserting the CMS solenoid

    CERN Multimedia

    Maximilien Brice

    2005-01-01

    The huge superconducting solenoid for CMS is inserted into the cryostat barrel. CMS uses the world's largest thin solenoid, in terms of energy stored, and is 12 m long, with a diameter of 6 m and weighing 220 tonnes. When turned on the magnet will produce a field strength of 4 T using superconducting niobium-titanium material at 4.5 K.

  3. Detection of chitinolytic enzymes with different substrate specificity in tissues of intact sundew (Drosera rotundifolia L.): chitinases in sundew tissues.

    Science.gov (United States)

    Libantová, Jana; Kämäräinen, Terttu; Moravcíková, Jana; Matusíková, Ildikó; Salaj, Jan

    2009-05-01

    The round-leaf sundew (Drosera rotundifolia L.) is a carnivorous plant expressing a wide range of chitinolytic enzymes playing role in many different processes. In this study the intact plants were analyzed for the presence of chitinase transcripts and chitinolytic activities in different organs. In situ hybridization with chitnase fragment as a probe has revealed the presence of chitinases in the mesophyll cells of leaves and vascular elements of stems of healthy, non-stressed plants. More pronounced expression was observed in cortex and stele cells of roots as well as in ovules and anthers of reproductive organs. Similarly, higher chitinase enzyme activity was typical for flowers and roots suggesting a more specific role of chitinases in these tissues. In addition to endochitinases of different substrate specificities, chitobiosidases contributed to overall chitinolytic activity of tissue extracts. The activity of chitobiosidases was again typical for flowers and roots, while their role in plant physiology remains to be elucidated. PMID:18437530

  4. Transformation of indica rice with plasmid pBGll21 containing a tobacco endo-chitinase gene I

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    @@ Several plasmids, which were suitable for cereals transformation, have been reported. In the study, rice was transformed by a new plasmid pBGll21 containing a tobacco endo-chitinase gene ( TchiB ).

  5. Contribution of Chitinase A’s C-Terminal Vacuolar Sorting Determinant to the Study of Soluble Protein Compartmentation

    Directory of Open Access Journals (Sweden)

    Egidio Stigliano

    2014-06-01

    Full Text Available Plant chitinases have been studied for their importance in the defense of crop plants from pathogen attacks and for their peculiar vacuolar sorting determinants. A peculiarity of the sequence of many family 19 chitinases is the presence of a C-terminal extension that seems to be important for their correct recognition by the vacuole sorting machinery. The 7 amino acids long C-terminal vacuolar sorting determinant (CtVSD of tobacco chitinase A is necessary and sufficient for the transport to the vacuole. This VSD shares no homology with other CtVSDs such as the phaseolin’s tetrapeptide AFVY (AlaPheValTyr and it is also sorted by different mechanisms. While a receptor for this signal has not yet been convincingly identified, the research using the chitinase CtVSD has been very informative, leading to the observation of phenomena otherwise difficult to observe such as the presence of separate vacuoles in differentiating cells and the existence of a Golgi-independent route to the vacuole. Thanks to these new insights in the endoplasmic reticulum (ER-to-vacuole transport, GFPChi (Green Fluorescent Protein carrying the chitinase A CtVSD and other markers based on chitinase signals will continue to help the investigation of vacuolar biogenesis in plants.

  6. Ozone and ultraviolet B effects on the defense-related proteins beta-1,3-glucanase and chitinase in tobacco

    International Nuclear Information System (INIS)

    The air pollutant ozone is a potent abiotic inducer of defense-related enzymes such as pathogenesis-related proteins. Here we report on the accumulation of beta-1,3-glucanase and chitinase in Nicotiana tabacumL. treated with ozone and ultraviolet B radiation, singly and in combination, under a simulated sunlight spectrum. Ozone (0.16 mu L . L-1, 2 x 5 h) induced the basic isoforms of beta-1,3-glucanase in both, ozone-sensitive (Eel W3) and -tolerant (Bel B) cultivars, while chitinase was only affected in cv. Bel W3. Ultraviolet B radiation (7.5 MED) alone did not lead to beta-1,3-glucanase or chitinase induction. In combined treatments ultraviolet B increased the ozone-dependent lesion formation and reduced chitinase accumulation in the sensitive cv. Bel W3. Analysis of the intercellular washing fluid of ozone-treated plants revealed the accumulation of a major ozone-related protein (O(3)R-1) of 28 kDa within 32 h. Microsequence analysis of two tryptic peptides showed 100 % homology to acidic chitinase PR-3b. These results indicate that basic beta-1,3-glucanase and chitinase are distinctly regulated in ozone and ultraviolet B treated tobacco, and that ultraviolet B radiation with a similar UV edge as the solar spectrum does not lead to an accumulation of basic pathogenesis-related proteins

  7. Two cold-induced family 19 glycosyl hydrolases from cherimoya (Annona cherimola) fruit: an antifungal chitinase and a cold-adapted chitinase.

    Science.gov (United States)

    Goñi, Oscar; Sanchez-Ballesta, María T; Merodio, Carmen; Escribano, María I

    2013-11-01

    Two cold-induced chitinases were isolated and purified from the mesocarp cherimoyas (Annona cherimola Mill.) and they were characterised as acidic endochitinases with a Mr of 24.79 and 47.77kDa (AChi24 and AChi48, respectively), both family 19 glycosyl hydrolases. These purified chitinases differed significantly in their biochemical and biophysical properties. While both enzymes had similar optimal acidic pH values, AChi24 was enzymatically active and stable at alkaline pH values, as well as displaying an optimal temperature of 45°C and moderate thermostability. Kinetic studies revealed a great catalytic efficiency of AChi24 for oligomeric and polymeric substrates. Conversely, AChi48 hydrolysis showed positive co-operativity that was associated to a mixture of different functional oligomeric states through weak transient protein interactions. The rise in the AChi48 kcat at increasing enzyme concentrations provided evidence of its oligomerisation. AChi48 chitinase was active and stable in a broad acidic pH range, and while it was relatively labile as temperatures increased, with an optimal temperature of 35°C, it retained about 50% of its maximal activity from 5 to 50°C. Thermodynamic characterisation reflected the high kcat of AChi48 and the remarkably lower ΔH(‡), ΔS(‡) and ΔG(‡) values at 5°C compared to AChi24, indicating that the hydrolytic activity of AChi48 was less thermodependent. In vitro functional studies revealed that AChi24 had a strong antifungal defence potential against Botrytis cinerea, whereas they displayed no cryoprotective or antifreeze activity. Hence, based on biochemical, thermodynamic and functional data, this study demonstrates that two acidic endochitinases are induced at low temperatures in a subtropical fruit, and that one of them acts in an oligomeric cold-adapted manner. PMID:23890591

  8. X-Ray Crystal Structure of the Full Length Human Chitotriosidase (CHIT1) Reveals Features of Its Chitin Binding Domain

    Science.gov (United States)

    Fadel, Firas; Zhao, Yuguang; Cousido-Siah, Alexandra; Ruiz, Francesc X.; Mitschler, André; Podjarny, Alberto

    2016-01-01

    Chitinases are enzymes that catalyze the hydrolysis of chitin. Human chitotriosidase (CHIT1) is one of the two active human chitinases, involved in the innate immune response and highly expressed in a variety of diseases. CHIT1 is composed of a catalytic domain linked by a hinge to its chitin binding domain (ChBD). This latter domain belongs to the carbohydrate-binding module family 14 (CBM14 family) and facilitates binding to chitin. So far, the available crystal structures of the human chitinase CHIT1 and the Acidic Mammalian Chitinase (AMCase) comprise only their catalytic domain. Here, we report a crystallization strategy combining cross-seeding and micro-seeding cycles which allowed us to obtain the first crystal structure of the full length CHIT1 (CHIT1-FL) at 1.95 Å resolution. The CHIT1 chitin binding domain (ChBDCHIT1) structure shows a distorted β-sandwich 3D fold, typical of CBM14 family members. Accordingly, ChBDCHIT1 presents six conserved cysteine residues forming three disulfide bridges and several exposed aromatic residues that probably are involved in chitin binding, including the highly conserved Trp465 in a surface- exposed conformation. Furthermore, ChBDCHIT1 presents a positively charged surface which may be involved in electrostatic interactions. Our data highlight the strong structural conservation of CBM14 family members and uncover the structural similarity between the human ChBDCHIT1, tachycitin and house mite dust allergens. Overall, our new CHIT1-FL structure, determined with an adapted crystallization approach, is one of the few complete bi-modular chitinase structures available and reveals the structural features of a human CBM14 domain. PMID:27111557

  9. MECHANISM AND HYDROPHOBIC FORCES DRIVING MEMBRANE PROTEIN INSERTION OF SUBUNIT II OF CYTOCHROME BO OXIDASE

    Science.gov (United States)

    Celebi, Nil; Dalbey, Ross E.; Yuan, Jijun

    2009-01-01

    Subunit II (CyoA) of cytochrome bo oxidase, which spans the inner membrane twice in bacteria, has several unusual features in membrane biogenesis. It is synthesized with an amino-terminal signal peptide. In addition, distinct pathways are used to insert the two ends of the protein. The amino-terminal domain is inserted by the YidC pathway whereas the large carboxyl-terminal domain is translocated by the SecYEG pathway. Insertion of the protein is also pmf-independent. In this study we examined the topogenic requirements and mechanism of insertion of CyoA in bacteria. We find that both the signal peptide and the first membrane spanning region are required for insertion of the amino-terminal periplasmic loop. The pmf-independence of insertion of the first periplasmic loop is due to the loop’s neutral net charge. We observe also that the introduction of negatively charged residues into the periplasmic loop makes insertion pmf dependent, whereas the addition of positively charged residues prevents insertion unless the pmf is abolished. Insertion of the carboxyl-terminal domain in the full-length CyoA occurs by a sequential mechanism even when the CyoA amino and carboxyl-terminal domains are swapped with other domains. However, when a long spacer peptide is added to increase the distance between the amino-terminal and carboxyl-terminal domains, insertion no longer occurs by a sequential mechanism. PMID:18155041

  10. Draft Genome Sequence of a Chitinase-producing Biocontrol Bacterium Serratia sp. C-1

    Directory of Open Access Journals (Sweden)

    Seur Kee Park

    2015-09-01

    Full Text Available The chitinase-producing bacterial strain C-1 is one of the key chitinase-producing biocontrol agents used for effective bioformulations for biological control. These bioformulations are mixed cultures of various chitinolytic bacteria. However, the precise identification, biocontrol activity, and the underlying mechanisms of the strain C-1 have not been investigated so far. Therefore, we evaluated in planta biocontrol efficacies of C-1 and determined the draft genome sequence of the strain in this study. The bacterial C-1 strain was identified as a novel Serratia sp. by a phylogenic analysis of its 16S rRNA sequence. The Serratia sp. C-1 bacterial cultures showed strong in planta biocontrol efficacies against some major phytopathogenic fungal diseases. The draft genome sequence of Serratia sp. C-1 indicated that the C-1 strain is a novel strain harboring a subset of genes that may be involved in its biocontrol activities.

  11. Potential of Chitinases as a Biopesticide against Agriculturally Harmful Fungi and Insects.

    Directory of Open Access Journals (Sweden)

    Gursharan Singh

    2013-12-01

    Full Text Available Due to an increasing sensibility and pressure of public and environmental agencies against the application of chemical based pesticides and their long lasting adverse effects on ecosystems and human health, has motivated the search for non hazardous alternatives. The most trustworthy substitute of chemical pesticides is considered as biocontrol agents. These agents could be formulations of bacteria, fungi, viruses, plant extracts or antibiotics. Inhibition or killing of harmful pests by biocontrol agents is environmentally safe and without creating any soil pollution. In last two decades, chitinases have received the attention of researchers for their anti-insects and antifungal biocontrol activities. This review critically focused on the successful studies (inside and outside of laboratory has been done for the evaluation of biocontrol potential of chitinases from different sources.

  12. ISOLATION AND CHARACTERIZATION OF CHITINASE GENE FROM THE UNTRADITIONAL PLANT SPECIES

    Directory of Open Access Journals (Sweden)

    Dominika Ďurechová

    2013-02-01

    Full Text Available Round-leaf sundew (Drosera rotundifolia L. from Droseraceae family belongs among a few plant species with strong antifungal potential. It was previously shown that chitinases of carnivorous plant species may play role during the insect prey digestion, when hard chitin skeleton is being decomposed. As many phytopathogenic fungi contain chitin in their cell wall our attention in this work was focused on isolation and in silico characterization of genomic DNA sequence of sundew chitinase gene. Subsequently this gene was fused to strong constitutive CaMV35S promoter and cloned into the plant binary vector pBinPlus and tested in A. tumefaciens LBA 4404 for its stability. Next, when transgenic tobacco plants are obtained, increasing of their antifungal potential will be tested.

  13. Role of Tyr-435 of Vibrio harveyi chitinase A in chitin utilization.

    Science.gov (United States)

    Sritho, Natchanok; Suginta, Wipa

    2012-03-01

    Vibrio harveyi chitinase A or VhChiA (EC.3.2.1.14) is a member of GH-18 chitinases that catalyzes chitin degradation from marine biomaterials. Our earlier structural data of VhChiA suggested that Tyr-435 marks the ending of subsite +2 and may influence binding of the interacting substrate at the aglycone binding sites. This study reports the effects of Tyr-435 using site-directed mutagenesis technique. Mutation of Tyr-435 to Ala (mutant Y435A) enhanced both binding and catalytic efficiency of VhChiA, whereas substitution of Tyr-435 to Trp (mutant Y435W) lessened the ability of the enzyme to bind and hydrolyze chitin substrates. The increased activity of Y435A can be explained by partial removal of a steric clash around subsite (+2), thereby allowing a chitin chain to move beyond or to access the enzyme's active site from the aglycone side more straightforwardly. PMID:22194054

  14. Chitinase-3-like-1/YKL-40 as marker of circulating tumor cells

    OpenAIRE

    Hamilton, Gerhard; Rath, Barbara; Burghuber, Otto

    2015-01-01

    Ex vivo expansion of circulating tumor cells (CTCs) of small cell lung cancer (SCLC) patients enabled systematic screening of secreted cytokines. Permanent CTC cultures of different patients shared secretion of chitinase-3-like-1 (CHI3L1)/YKL-40, known to be upregulated in a range of tumor entities and to be associated with increased metastasis and decreased survival. This protein lacks enzymatic activity and its mechanism of promoting tumor dissemination has not been resolved. Results from S...

  15. Effect of Chitinase-Producing Strain V-8 on 3ontrolling Cotton Fusarium Wilt

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    [Objective] This study aimed to screen endophytic bacteria which is antag- onistic to cotton Fusarium wilt. [Method] Fresh cotton plants collected from cotton- growing areas in Jingzhou City, Hubei Province were used as experimental materials to isolate endophytic bacteria. Through chitinase test and co-culturing both micro-or- ganisms side by side on the same PDA culture plate, antagonistic strains to cotton Fusarium wilt were screened. [Result] A total of 83 bacterial isolates were obtained from cotton plants grown in the fields, six of which were chitinase-productive bacte- ria. Through chitinase test and co-culturing both micro-organisms side by side on the same PDA culture plate, strain V-8 which had the strongest antagonistic effect on Fusarium oxysporum f. sp. vasinfectum was screened. Strain V-8 had a wider anti- fungal spectrum with certain inhibitory effect on all the six important pathogenic fungi including Fusarium oxysporum f. sp niveum; it colonized stably in the rhizospheric soil of cotton, with a colonization density of up to 6.2x10s cfu/g fifty days after inoc- ulation; the relative effect on controlling cotton Fusarium wilt in pot test was 73.2%. The Findings of this study suggested that strain V-8 had great potential for biological control of cotton Fusarium wilt and could be taken as a substantial material for the cloning of chitinase genes. [Conclusion] The results from this study provides bases for the control of cotton fusarium wilt, as well as the exploitation of endophytic bac- teria resources in cotton and the development of novel biological pesticides.

  16. Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer

    Directory of Open Access Journals (Sweden)

    Sebastian Margino

    2015-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Potato Cyst Nematodes (Globodera rostochiensis is one of the important potato’s pests and caused economic looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN shell component of egg shell containing chitin (inner layer and vitelline/protein (outer layer, so the purpose of research was to fi nd out of chitin degrading bacteria for controlling of egg’s PCN by cutting of their life cycle. The results showed that Bacillus sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation using ammonium sulphate precipitation and DEAE-Cellulose ion-exchange chromatography showed a specifi c activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The apparent Km and Vmax of chitinase towards colloidal chitin were 2 mg/ml and 2.2 μg/h, respectively.  

  17. Purification and characterization of a novel chitinase from Trichosanthes dioica seed with antifungal activity.

    Science.gov (United States)

    Kabir, Syed Rashel; Rahman, Md Musfikur; Tasnim, Shahnima; Karim, Md Rezaul; Khatun, Nazma; Hasan, Imtiaj; Amin, Ruhul; Islam, Shaikh Shohidul; Nurujjaman, Md; Kabir, Ahmad Humayan; Sana, Niranjan Kumar; Ozeki, Yasuhiro; Asaduzzaman, A K M

    2016-03-01

    Chitinases are a group of enzymes that show differences in their molecular structure, substrate specificity, and catalytic mechanism and widely found in organisms like bacteria, yeasts, fungi, arthropods actinomycetes, plants and humans. A novel chitinase enzyme (designated as TDSC) was purified from Trichosanthes dioica seed with a molecular mass of 39±1 kDa in the presence and absence of β-mercaptoethanol. The enzyme was a glycoprotein in nature containing 8% neutral sugar. The N-terminal sequence was determined to be EINGGGA which did not match with other proteins. Amino acid analysis performed by LC-MS revealed that the protein was rich in leucine. The enzyme was stable at a wide range of pH (5.0-11.0) and temperature (30-90 °C). Chitinase activity was little bit inhibited in the presence of chelating agent EDTA (ethylenediaminetetraaceticacid), urea and Ca(2+). A strong fluorescence quenching effect was found when dithiothreitol and sodium dodecyl sulfate were added to the enzyme. TDSC showed antifungal activity against Aspergillus niger and Trichoderma sp. as tested by MTT assay and disc diffusion method. PMID:26666429

  18. Chitinase production and antifungal potential of endophytic Streptomyces strain P4

    Directory of Open Access Journals (Sweden)

    Hataichanoke Niamsup

    2012-02-01

    Full Text Available The endophytic actinomycete P4 strain, previously isolated from sweet pea root, wasidentified as Streptomyces sp. by full 16S rRNA sequencing. It is mostly related to Streptomycesgriseoflavus with a 99.7% identity score. The Streptomyces sp. P4 was tested for its hydrolyticactivities by plate method. The result showed the presence of chitinase. The extent of chitinase activitywas assessed by spectrophotometric method along with growth monitoring. Chitinase production wasgrowth-associated and showed the highest activity on the fifth day. The dual culture method revealedthat the strain was effective in restricting the radial growth of Fusarium oxysporum f.sp. lycopersici, animportant phytopathogen of tomato. Scanning electronic microscopic analysis showed that the ruptureof the F. oxysporum mycelial cell wall occurred at the area of interaction between F. oxysporum andStreptomyces sp. P4. This was possibly due to the chitinolytic activity of the P4. Thus, thisactinomycete has the potential for being used as a biocontrol agent, thereby reducing the use ofchemical fungicides.

  19. Purification and characterization of an antifungal chitinase from Bacillus sp.SL-13

    Institute of Scientific and Technical Information of China (English)

    Chen; Shan

    2014-01-01

    Bacillus sp.SL-13 produced antifungal proteins.The growth of the plant-pathogenic fungi Rhizoctonia solani was considerably inhibited by the presence of the SL-13 culture supernatant.The proteins were purified by DEAE-Sepharose fast flow ion exchange column chromatography and Sephadex G-75 gel filtration,and the main antifungal protein was purified to be chitinase.The molecular weight of chitinase was estimated to be 36 kD by 12%SDS PAGE.The optimal pH and temperature for the chitinase was 7.0 and 50℃.It demonstrated that the enzyme was stable from pH 5 to 9 and form 40?C to 60℃.The enzyme still kept 70%activity when incubated at 121℃,0.11MPa up to 20 minutes and the enzyme is also not lost the activity when treated with protease K and ultraviolet radiation for 1.5hours.It is very suitable for the use in a relatively unstable environment,exhibiting effective biological control.

  20. Optimization of Chitinase Production by Bacillus pumilus Using Plackett-Burman Design and Response Surface Methodology

    Science.gov (United States)

    Tasharrofi, Noshin; Adrangi, Sina; Fazeli, Mehdi; Rastegar, Hossein; Khoshayand, Mohammad Reza; Faramarzi, Mohammad Ali

    2011-01-01

    A soil bacterium capable of degrading chitin on chitin agar plates was isolated and identified as Bacillus pumilus isolate U5 on the basis of 16S rDNA sequence analysis. In order to optimize culture conditions for chitinase production by this bacterium, a two step approach was employed. First, the effects of several medium components were studied using the Plackett-Burman design. Among various components tested, chitin and yeast extract showed positive effect on enzyme production while MgSO4 and FeSO4 had negative effect. However, the linear model proved to be insufficient for determining the optimum levels for these components due to a highly significant curvature effect. In the second step, Box-Behnken response surface methodology was used to determine the optimum values. It was noticed that a quadratic polynomial equation fitted he experimental data appropriately. The optimum concentrations for chitin, yeast extract, MgSO4 and FeSO4 were found to be 4.76, 0.439, 0.0055 and 0.019 g/L, respectively, with a predicted value of chitinase production of 97.67 U/100 mL. Using this statistically optimized medium, the practical chitinase production reached 96.1 U/100 mL. PMID:24250411

  1. Bacterial and fungal chitinase chiJ orthologs evolve under different selective constraints following horizontal gene transfer

    Directory of Open Access Journals (Sweden)

    Ubhayasekera Wimal

    2012-10-01

    Full Text Available Abstract Background Certain bacteria from the genus Streptomyces are currently used as biological control agents against plant pathogenic fungi. Hydrolytic enzymes that degrade fungal cell wall components, such as chitinases, are suggested as one possible mechanism in biocontrol interactions. Adaptive evolution of chitinases are previously reported for plant chitinases involved in defence against fungal pathogens, and in fungal chitinases involved in fungal-fungal interactions. In this study we investigated the molecular evolution of chitinase chiJ in the bacterial genus Streptomyces. In addition, as chiJ orthologs are previously reported in certain fungal species as a result from horizontal gene transfer, we conducted a comparative study of differences in evolutionary patterns between bacterial and fungal taxa. Findings ChiJ contained three sites evolving under strong positive selection and four groups of co-evolving sites. Regions of high amino acid diversity were predicted to be surface-exposed and associated with coil regions that connect certain α-helices and β-strands in the family 18 chitinase TIM barrel structure, but not associated with the catalytic cleft. The comparative study with fungal ChiJ orthologs identified three regions that display signs of type 1 functional divergence, where unique adaptations in the bacterial and fungal taxa are driven by positive selection. Conclusions The identified surface-exposed regions of chitinase ChiJ where sequence diversification is driven by positive selection may putatively be related to functional divergence between bacterial and fungal orthologs. These results show that ChiJ orthologs have evolved under different selective constraints following the horizontal gene transfer event.

  2. ENDOSCOPIC GROMMET INSERTION OUR EXPERIENCE

    Directory of Open Access Journals (Sweden)

    Balasubramanian Thiagarajan

    2012-03-01

    Full Text Available Grommet insertion the commonest surgical procedure next only to circumcision is usually performed using an operating microscope 1. Authors have been using 4 mm 0 degree nasalendoscopes to perform this procedure during the last 5 years. This is a report of their experience in using endoscope inlieu of microscope in performing this surgery. This study makes a comparative analysis of Endoscopic Grommet insertion viz a viz Microscopic Grommet insertion. For this comparative analysis one year (2009 data base of Government Stanley Medical College Chennai India was used. This study reveals that Endoscopic Grommet insertion compared favorably with Microscopic Grommet insertion in all aspects with certain obvious advantages.

  3. Antifungal performance of extracellular chitinases and culture supernatants of Streptomyces galilaeus CFFSUR-B12 against Mycosphaerella fijiensis Morelet.

    Science.gov (United States)

    Castillo, Benjamín Moreno; Dunn, Michael F; Navarro, Karina Guillén; Meléndez, Francisco Holguín; Ortiz, Magdalena Hernández; Guevara, Sergio Encarnación; Palacios, Graciela Huerta

    2016-03-01

    The tropical and mycoparasite strain Streptomyces galilaeus CFFSUR-B12 was evaluated as an antagonist of Mycosphaerella fijiensis Morelet, causal agent of the Black Sigatoka Disease (BSD) of banana. On zymograms of CFFSUR-B12 culture supernatants, we detected four chitinases of approximately 32 kDa (Chi32), 20 kDa (Chi20), and two with masses well over 170 kDa (ChiU) that showed little migration during denaturing electrophoresis at different concentrations of polyacrylamide. The thymol-sulphuric acid assay showed that the ChiU were glycosylated chitinases. Moreover, matrix assisted laser desorption ionization time-of-flight MS analysis revealed that the ChiU are the same protein and identical to a family 18 chitinase from Streptomyces sp. S4 (gi|498328075). Chi32 was similar to an extracellular protein from Streptomyces albus J1074 (gi|478687481) and Chi20 was non-significantly similar to chitinases from five different strains of Streptomyces (P > 0.05). Subsequently, Chi32 and Chi20 were partially purified by anion exchange and hydrophobic interaction chromatography and tested against M. fijiensis. Chitinases failed to inhibit ascospore germination, but inhibited up to 35 and 62 % of germ tube elongation and mycelial growth, respectively. We found that crude culture supernatant and living cells of S. galilaeus CFFSUR-B12 were the most effective in inhibiting M. fijiensis and are potential biocontrol agents of BSD. PMID:26873555

  4. Development of insect resistant maize plants expressing a chitinase gene from the cotton leaf worm, Spodoptera littoralis.

    Science.gov (United States)

    Osman, Gamal H; Assem, Shireen K; Alreedy, Rasha M; El-Ghareeb, Doaa K; Basry, Mahmoud A; Rastogi, Anshu; Kalaji, Hazem M

    2015-01-01

    Due to the importance of chitinolytic enzymes for insect, nematode and fungal growth, they are receiving attention concerning their development as biopesticides or chemical defense proteins in transgenic plants and as microbial biocontrol agents. Targeting chitin associated with the extracellular matrices or cell wall by insect chitinases may be an effective approach for controlling pest insects and pathogenic fungi. The ability of chitinases to attack and digest chitin in the peritrophic matrix or exoskeleton raises the possibility to use them as insect control method. In this study, an insect chitinase cDNA from cotton leaf worm (Spodoptera littoralis) has been synthesized. Transgenic maize plant system was used to improve its tolerance against insects. Insect chitinase transcripts and proteins were expressed in transgenic maize plants. The functional integrity and expression of chitinase in progenies of the transgenic plants were confirmed by insect bioassays. The bioassays using transgenic corn plants against corn borer (Sesamia cretica) revealed that ~50% of the insects reared on transgenic corn plants died, suggesting that transgenic maize plants have enhanced resistance against S. cretica. PMID:26658494

  5. Cloning and identification of Fv-cmp, a protease from Fusarium verticillioides that truncates Zea mays and Arabidopsis thaliana class IV chitinases

    Science.gov (United States)

    Chitinase modifying proteins (cmps) are proteases, secreted by fungal pathogens, that were originally identified as proteins that truncate class IV chitinases of maize during ear rot. Cmps from Bipolaris zeicola and Stenocarpella maydis have been characterized, but the identities of the proteases h...

  6. Facility target insert shielding assessment

    Energy Technology Data Exchange (ETDEWEB)

    Mocko, Michal [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2015-10-06

    Main objective of this report is to assess the basic shielding requirements for the vertical target insert and retrieval port. We used the baseline design for the vertical target insert in our calculations. The insert sits in the 12”-diameter cylindrical shaft extending from the service alley in the top floor of the facility all the way down to the target location. The target retrieval mechanism is a long rod with the target assembly attached and running the entire length of the vertical shaft. The insert also houses the helium cooling supply and return lines each with 2” diameter. In the present study we focused on calculating the neutron and photon dose rate fields on top of the target insert/retrieval mechanism in the service alley. Additionally, we studied a few prototypical configurations of the shielding layers in the vertical insert as well as on the top.

  7. Endoscopic grommet insertion our experience

    OpenAIRE

    Balasubramanian Thiagarajan; Venkatesan Ulaganathan; Seethalakshmi Narashiman

    2012-01-01

    Grommet insertion the commonest surgical procedure next only to circumcision is usually performed using an operating microscope 1. Authors have been using 4 mm 0 degree nasal endoscopes to perform this procedure during the last 5 years. This is a report of their experience in using endoscope inlieu of microscope in performing this surgery. This study makes a comparative analysis of Endoscopic Grommet insertion viz a viz Microscopic Grommet insertion. For this comparative analysis one year (20...

  8. Subjective Misidentification and Thought Insertion

    OpenAIRE

    Parrott, Matthew Thomas

    2015-01-01

    This essay presents a new account of thought insertion. Prevailing views in both philosophy and cognitive science tend to characterize the experience of thought insertion as missing or lacking some element, such as a ‘sense of agency’, found in ordinary first-person awareness of one’s own thoughts. By contrast, I propose that, rather than lacking something, experiences of thought insertion have an additional feature not present in ordinary conscious experiences of one’s own thoughts. More spe...

  9. Optimization of nutrition factors on chitinase production from a newly isolated Chitiolyticbacter meiyuanensis SYBC-H1

    Directory of Open Access Journals (Sweden)

    Zhikui Hao

    2012-03-01

    Full Text Available The present study reports statistical medial optimization for chitinase production by a novel bacterial strain isolated from soil recently, which the name Chitinolyticbacter meiyuanensis SYBC-H1 is proposed. A sequential statistical methodology comprising of Plackett-Burman and response surface methodology (RSM was applied to enhance the fermentative production of chitinase, in which inulin was firstly used as an effective carbon source. As a result, maximum chitinase activity of 5.17 U/mL was obtained in the optimized medium, which was 15.5-fold higher than that in the basal medium. The triplicate verification experiments were performed under the optimized nutrients levels which indicated that it well agreed with the predicted value.

  10. Post-CTS Delay Insertion

    OpenAIRE

    Jianchao Lu; Baris Taskin

    2010-01-01

    A post-clock-tree-synthesis (post-CTS) optimization method is proposed that suggests delay insertion at the leaves of the clock tree in order to implement a limited version of clock skew scheduling. Delay insertion is limited on each clock tree branch simultaneous with a global monitoring of the total amount of delay insertion. The delay insertion for nonzero clock skew operation is performed only at the clock sinks in order to preserve the structure and the optimizations implemented in the c...

  11. Cloning of a chitinase gene from Ewingella americana, a pathogen of the cultivated mushroom, Agaricus bisporus

    Directory of Open Access Journals (Sweden)

    P.W. Inglis

    2000-09-01

    Full Text Available We have isolated a gene encoding a chitinase (EC 3.2.1.14 from Ewingella americana, a recently described pathogen of the mushroom Agaricus bisporus. This gene, designated chiA (EMBL/Genbank/DDBJ accession number X90562, was cloned by expression screening of a plasmid-based E. americana HindIII genomic library in Escherichia coli using remazol brilliant violet-stained carboxymethylated chitin incorporated into selective medium. The chiA gene has a 918-bp ORF, terminated by a TAA codon, with a calculated polypeptide size of 33.2 kDa, likely corresponding to a previously purified and characterised 33-kDa endochitinase from E. americana. The deduced amino acid sequence shares 33% identity with chitinase II from Aeromonas sp. No. 10S-24 and 7.8% identity with a chitinase from Saccharopolyspora erythraeus. Homology to other chitinase sequences was otherwise low. The peptide sequence deduced from chiA lacks a typical N-terminal signal sequence and also lacks the chitin binding and type III fibronectin homology units common to many bacterial chitinases. The possibility that this chitinase is not primarily adapted for the environmental mineralisation of pre-formed chitin, but rather for the breakdown of nascent chitin, is discussed in the context of mushroom disease.O gene que codifica uma quitinase (EC 3.2.1.14 foi isolado de Ewingella americana, recentemente descrita como patógeno do cogumelo Agaricus bisporus. Este gene, denominado chiA (EMBL/Genebank/DDBJ número de acesso X9061, foi clonado e selecionado a partir de livraria genômica construída por digestão do DNA de E. americana com HindIII e ligação em plasmídio de expressão em E. coli, utilizando meio seletivo contendo quitina carboximetilada, corada com "remazol brilliant violet'' para seleção de clones. O gene chiA apresenta uma ORF de 918 bp, código terminador TAA, tendo o tamanho do polipeptídeo sido calculado como 33,2 kDa, o qual corresponde ao tamanho de 33 kDa da endoquitinase

  12. Production of prodigiosin and chitinases by tropical Serratia marcescens strains with potential to control plant pathogens.

    Science.gov (United States)

    Gutiérrez-Román, Martha Ingrid; Holguín-Meléndez, Francisco; Bello-Mendoza, Ricardo; Guillén-Navarro, Karina; Dunn, Michael F; Huerta-Palacios, Graciela

    2012-01-01

    The potential of three Serratia marcescens strains (CFFSUR-B2, CFFSUR-B3 and CFFSUR-B4) isolated from tropical regions in Mexico to inhibit the mycelial growth and conidial germination of Colletotrichum gloeosporioides, causal agent of fruit anthracnose, was evaluated. The ability of these strains to produce prodigiosin and chitinases when cultivated in oil seed-based media (peanut, sesame, soybean and castor bean) and in Luria-Bertani medium was determined. All of the strains exhibited similar fungal antagonistic activities and inhibited myceliar growth by more than 40% while inhibiting conidial germination by 81-89% (P = 0.01). The highest level of prodigiosin (40 μg/ml) was produced in the peanut-based medium while growth in soybean-based medium allowed the highest production of chitinases (56 units/ml), independent of the strain used. Strain CFFSUR-B2 grown in peanut medium was used to evaluate the effect of inoculum density and initial pH on metabolite production. The amount of prodigiosin produced increased with greater inoculum densities, with an initial density of 1 × 10(12) resulting in the highest production (60 μg/ml). Prodigiosin production was not affected by pH. The strains studied have the advantage of being adapted to tropical climates and are able to produce chitinases in the absence of chitin induction in vitro. These characteristics suggest their potential as biocontrol agents for fungal pathogens in tropical regions of the world. PMID:22806790

  13. Purification, characterization and structural determination of chitinases produced by Moniliophthora perniciosa

    OpenAIRE

    Rafaela S. Galante; Taranto, Alex G; Maria G.B. Koblitz; Aristóteles Góes-Neto; Pirovani, Carlos P.; Cascardo, Júlio C. M.; Sandra H. Cruz; Pereira, Gonçalo A. G.; Assis, Sandra A

    2012-01-01

    The enzyme chitinase from Moniliophthora perniciosa the causative agent of the witches' broom disease in Theobroma cacao, was partially purified with ammonium sulfate and filtration by Sephacryl S-200 using sodium phosphate as an extraction buffer. Response surface methodology (RSM) was used to determine the optimum pH and temperature conditions. Four different isoenzymes were obtained: ChitMp I, ChitMp II, ChitMp III and ChitMp IV. ChitMp I had an optimum temperature at 44-73ºC and an optimu...

  14. Isolation of Serratia marcescens SR1 as a Source of Chitinase Having Potentiality of Using as a Biocontrol Agent

    OpenAIRE

    Parani, K.; Shetty, G. P.; Saha, B K

    2011-01-01

    Serratia marcescens, strain SR1 was isolated from the local soil of a cultivated farm and it was screened as potent strain for chitinase production. Maximum chitinase production (77.3 u Mh−1 100−1) was observed after 96 h of incubation period with pH 5.5 at 30°C under shake conditions (120 rpm). Compare to still flasks, shake culture with prawn fish colloidal chitin of 0.5% (w/v) concentration, showed a better enzyme yield. Crude enzyme showed antifungal activity against plant pathogens....

  15. Molecular cloning, expression and biochemical characterisation of a cold-adapted novel recombinant chitinase from Glaciozyma antarctica PI12

    Directory of Open Access Journals (Sweden)

    Ramli Aizi

    2011-11-01

    Full Text Available Abstract Background Cold-adapted enzymes are proteins produced by psychrophilic organisms that display a high catalytic efficiency at extremely low temperatures. Chitin consists of the insoluble homopolysaccharide β-(1, 4-linked N-acetylglucosamine, which is the second most abundant biopolymer found in nature. Chitinases (EC 3.2.1.14 play an important role in chitin recycling in nature. Biodegradation of chitin by the action of cold-adapted chitinases offers significant advantages in industrial applications such as the treatment of chitin-rich waste at low temperatures, the biocontrol of phytopathogens in cold environments and the biocontrol of microbial spoilage of refrigerated food. Results A gene encoding a cold-adapted chitinase (CHI II from Glaciozyma antarctica PI12 was isolated using Rapid Amplification of cDNA Ends (RACE and RT-PCR techniques. The isolated gene was successfully expressed in the Pichia pastoris expression system. Analysis of the nucleotide sequence revealed the presence of an open reading frame of 1,215 bp, which encodes a 404 amino acid protein. The recombinant chitinase was secreted into the medium when induced with 1% methanol in BMMY medium at 25°C. The purified recombinant chitinase exhibited two bands, corresponding to the non-glycosylated and glycosylated proteins, by SDS-PAGE with molecular masses of approximately 39 and 50 kDa, respectively. The enzyme displayed an acidic pH characteristic with an optimum pH at 4.0 and an optimum temperature at 15°C. The enzyme was stable between pH 3.0-4.5 and was able to retain its activity from 5 to 25°C. The presence of K+, Mn2+ and Co2+ ions increased the enzyme activity up to 20%. Analysis of the insoluble substrates showed that the purified recombinant chitinase had a strong affinity towards colloidal chitin and little effect on glycol chitosan. CHI II recombinant chitinase exhibited higher Vmax and Kcat values toward colloidal chitin than other substrates at low

  16. Insertion device vacuum system designs

    International Nuclear Information System (INIS)

    Synchrotron light source insertion device vacuum systems now in operation and systems proposed for the future are reviewed. An overview of insertion devices is given and four generic vacuum chamber designs, transition section design and pumping considerations are discussed. Examples of vacuum chamber systems are presented

  17. ISR Superconducting High luminosity Insertion

    CERN Multimedia

    1981-01-01

    The picture shows two of the eight superconducting quadrupoles of the low-beta insertion at intersection I8.The increase of luminosity produced by this insertion was above a factor 7. At right one can also see the Open- Axial- Field Magnet. The person is Stephan Pichler. See also 7702690X, 8102123, 8010397, 8008332.

  18. Agrobacterium mediated transformation of brassica juncea (l.) czern with chitinase gene conferring resistance against fungal infections

    International Nuclear Information System (INIS)

    Brassica juncea (Czern and Coss., L.) is an important oilseed crop. Since it is attacked by several bacterial and fungal diseases, therefore, we developed an easy and simple protocol for the regeneration and transformation of B. juncea variety RAYA ANMOL to give rise to transgenic plants conferring resistance against various fungal diseases. The transformation was carried out using Agrobacterium with Chitinase gene. This gene was isolated from Streptomyces griseus HUT6037. We used two types of explants for transformation i.e. hypocotyls and cotyledons. Only hypocotyls explants showed good results regarding callus initiation. Different hormonal concentrations were applied i.e. BAP 2, 4 and 6 mgL-1 and NAA 0.1, 0.2 and 0.3 mgL-1. However, high transformation efficiency was observed by supplementing the medium with combination of 2 mgL-1 BAP and 0.2 mgL-1 for initiation of callus. Similarly 10 mgL-1 kanamycin and 200 mgL-1 cefotaxime also proved successful for the selection of transformed callus. In order to confirm the presence of transgenic callus Polymerase chain reaction was performed using specific primers for Chitinase gene. (author)

  19. Preparation of nanoscale Bacillus thuringiensis chitinases using silica nanoparticles for nematicide delivery.

    Science.gov (United States)

    Qin, Xu; Xiang, Xuemei; Sun, Xiaowen; Ni, Hong; Li, Lin

    2016-01-01

    A series of amino, carboxylic, and aldehydic surface-grafted silica nanoparticles (SNPs) was prepared based on SiO2 NYSi40 nanoparticles to develop an efficient, biocompatible, and cost-effective biopesticide delivery system. Bacillus thuringiensis chitinase (Chi9602) was immobilized onto SNP surface to prepare nanoscale chitinases (SNPCs) through electrostatic adsorption and covalent binding. The specimens were characterized by Fourier transform infrared, scanning electron microscopy, and zeta-potential analyses. The delivery capacity of the SNPs in Caenorhabditis elegans N2 was observed by immunofluorescence. Results demonstrated that amino-grafted SiO2 nanoparticles with Chi9602 electrostatically adsorbed onto their surface (SNPC2) exhibited a relatively high enzyme immobilization rate (80.2%) and the highest (94.1%) residual enzyme activity among all SNPCs. SNPC2 also showed wider pH tolerance and relatively higher thermostability and ultraviolet radiation resistance capacity than Chi9602. Bioassays further showed that SNPC2 synergistically enhanced the nematicidal effect of B. thuringiensis YBT-020 preparation against C. elegans, with a reduced LC50 of 8.35mg/mL and a shortened LT50 of 12.04h. Immunofluorescence assays showed that SNPC2 had considerable delivery capacity to carry a large protein into C. elegans. Therefore, SNP2 can serve as an efficient nanocarrier for the delivery of macromolecular proteic biopesticides or drugs, indicating potential agricultural or biotechnological applications. PMID:26476241

  20. Expression, purification, crystallization and preliminary crystallographic analysis of chitinase A from Vibrio carchariae

    International Nuclear Information System (INIS)

    Chitinase A of Vibrio carchariae was functionally expressed in Escherichia coli M15 host cells as a C-terminally proteolytic processed fragment using the pQE60 expression vector. The yield of the 63-kDa protein was purified, yielding ∼70 mg per liter of bacterial culture. Crystals of recombinant chitinase A were obtained by the hanging-drop vapor diffusion method in a precipitant containing 10% (v/v) PEG 400, 0.1 M sodium acetate p H 4.6 and 0.125 M CaCl2. The crystals belonged to the tetragonal space group P422 with two molecules per asymmetric unit and unit-cell parameters a = b 127.64 Angstrom, and c = 171.42 Angstrom. A complete diffraction data set was collected to 2.14 Angstrom resolution, using a Rigaku/MSC R-AXIS IV++ detector system mounted on an RU-H3R rotating-anode X-ray generator

  1. Purification and characterization of a viral chitinase active against plant pathogens and herbivores from transgenic tobacco.

    Science.gov (United States)

    Di Maro, Antimo; Terracciano, Irma; Sticco, Lucia; Fiandra, Luisa; Ruocco, Michelina; Corrado, Giandomenico; Parente, Augusto; Rao, Rosa

    2010-05-01

    The Autographa californica nucleopolyhedrovirus chitinase A (AcMNPV ChiA) is a chitinolytic enzyme with fungicidal and insecticidal properties. Its expression in transgenic plants enhances resistance against pests and fungal pathogens. We exploited tobacco for the production of a biologically active recombinant AcMNPV ChiA (rChiA), as such species is an alternative to traditional biological systems for large-scale enzyme production. The protein was purified from leaves using ammonium sulfate precipitation followed by anion exchange and gel-filtration chromatography. Transgenic plants produced an estimated 14 mg kg(-1) fresh leaf weight, which represents 0.2% of total soluble proteins. The yield of the purification was about 14% (2 mg kg(-1) fresh leaf weight). The comparison between the biochemical and kinetic properties of the rChiA with those of a commercial Serratia marcescens chitinase A indicated that the rChiA was thermostable and more resistant at basic pH, two positive features for agricultural and industrial applications. Finally, we showed that the purified rChiA enhanced the permeability of the peritrophic membrane of larvae of two Lepidoptera (Bombyx mori and Heliothis virescens) and inhibited spore germination and growth of the phytopatogenic fungus Alternaria alternata. The data indicated that tobacco represents a suitable platform for the production of rChiA, an enzyme with interesting features for future applications as "eco-friendly" control agent in agriculture. PMID:20302895

  2. Chitinase 3-like 1 Regulates Cellular and Tissue Responses via IL-13 Receptor α2

    Directory of Open Access Journals (Sweden)

    Chuan Hua He

    2013-08-01

    Full Text Available Members of the 18 glycosyl hydrolase (GH 18 gene family have been conserved over species and time and are dysregulated in inflammatory, infectious, remodeling, and neoplastic disorders. This is particularly striking for the prototypic chitinase-like protein chitinase 3-like 1 (Chi3l1, which plays a critical role in antipathogen responses where it augments bacterial killing while stimulating disease tolerance by controlling cell death, inflammation, and remodeling. However, receptors that mediate the effects of GH 18 moieties have not been defined. Here, we demonstrate that Chi3l1 binds to interleukin-13 receptor α2 (IL-13Rα2 and that Chi3l1, IL-13Rα2, and IL-13 are in a multimeric complex. We also demonstrate that Chi3l1 activates macrophage mitogen-activated protein kinase, protein kinase B/AKT, and Wnt/β-catenin signaling and regulates oxidant injury, apoptosis, pyroptosis, inflammasome activation, antibacterial responses, melanoma metastasis, and TGF-β1 production via IL-13Rα2-dependent mechanisms. Thus, IL-13Rα2 is a GH 18 receptor that plays a critical role in Chi3l1 effector responses.

  3. Quantification of chitinase and thaumatin-like proteins in grape juices and wines.

    Science.gov (United States)

    Le Bourse, D; Conreux, A; Villaume, S; Lameiras, P; Nuzillard, J-M; Jeandet, P

    2011-09-01

    Chitinases and thaumatin-like proteins are important grape proteins as they have a great influence on wine quality. The quantification of these proteins in grape juices and wines, along with their purification, is therefore crucial to study their intrinsic characteristics and the exact role they play in wines. The main isoforms of these two proteins from Chardonnay grape juice were thus purified by liquid chromatography. Two fast protein liquid chromatography (FLPC) steps allowed the fractionation and purification of the juice proteins, using cation exchange and hydrophobic interaction media. A further high-performance liquid chromatography (HPLC) step was used to achieve higher purity levels. Fraction assessment was achieved by mass spectrometry. Fraction purity was determined by HPLC to detect the presence of protein contaminants, and by nuclear magnetic resonance (NMR) spectroscopy to detect the presence of organic contaminants. Once pure fractions of lyophilized chitinase and thaumatin-like protein were obtained, ultra-HPLC (UHPLC) and enzyme-linked immunosorbent assay (ELISA) calibration curves were constructed. The quantification of these proteins in different grape juice and wine samples was thus achieved for the first time with both techniques through comparison with the purified protein calibration curve. UHPLC and ELISA showed very consistent results (less than 16% deviation for both proteins) and either could be considered to provide an accurate and reliable quantification of proteins in the oenology field. PMID:21465097

  4. Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifolia L.) show induction of chitinase activity upon mimicking the presence of prey.

    Science.gov (United States)

    Matusíková, Ildikó; Salaj, Ján; Moravcíková, Jana; Mlynárová, Ludmila; Nap, Jan-Peter; Libantová, Jana

    2005-12-01

    Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addition, mechanical stirring of tentacles was performed. Chitinase activity was markedly increased in leaf exudates upon application of notably chitin. Application of gelatine increased the proteolytic activity of leaf exudates, indicating that the reaction of sundew leaves depends on the molecular nature of the inducer applied. In situ hybridization of sundew leaves with a Drosera chitinase probe showed chitinase gene expression in different cell types of non-treated leaves, but not in the secretory cells of the glandular heads. Upon induction, chitinase mRNA was also present in the secretory cells of the sundew leaf. The combined results indicate that chitinase is likely to be involved in the decomposition of insect prey by carnivorous plants. This adds a novel role to the already broad function of chitinases in the plant kingdom and may contribute to our understanding of the molecular mechanisms behind the ecological success of carnivorous plants in nutritionally poor environments. PMID:16049675

  5. Isolation and molecular identification chitinase-producing Streptomyces strains and examination of their in-vitro antagonistic effects

    Directory of Open Access Journals (Sweden)

    Alireza Dehnad

    2015-12-01

    Full Text Available Introduction: The chemical fungicides are used widely in the world. To reduce the application of synthetic fungicides in treating plant diseases, biological methods are considered as an alternative way to control plant diseases. Many actinomycetes, particularly Streptomyces species are biological agents against a broad spectrum of fungal plant pathogens. The purpose of this study was using the kitinolitik actinomycetes isolated from soil of Eastern Azerbaijan province In order to produce biological pesticides. Materials and methods: Soil samples were taken from different areas of Eastern Azerbaijan province. According to Streptomyces morphological features, single colonies were isolated. To identify the bacteria by molecular characteristic, the genomic DNA was extracted and then the sequences of 16S rDNA were replicated. By using specific primers the bacterial isolates containing chitinase gene were screened. The isolates consisted Chitinase enzyme and were antagonistically cultured with Alternaria genus which is a fungal plant pathogen. Results: Out of 60 soil collected samples, 31 Streptomyces bacterial isolates were separated. Four isolates showed positive results to selectivity action of the chitinase enzyme. Treatment of 3 bacterial isolates with 2 pathogenic fungi showed that AE09 is the most effective anti-fungal isolates. Discussion and conclusion: Soils in Eastern Azerbaijan province are rich of Streptomyces bacteria which generate antifungal compounds. Obtaining the Streptomyces bacteria which have chitinase gene, can lead to identification of very effective strains as anti-fungal.

  6. The Ifchit1 chitinase gene acts as a critical virulence factor in the insect pathogenic fungus Isaria fumosorosea.

    Science.gov (United States)

    Huang, Zhen; Hao, Yongfen; Gao, Tianni; Huang, Yü; Ren, Shunxiang; Keyhani, Nemat O

    2016-06-01

    The filamentous fungus, Isaria fumosorosea, is a promising insect biological control agent. Chitinases have been implicated in targeting insect cuticle structures, with biotechnological potential in insect and fungal control. The I. fumosorosea chitinase gene, Ifchit1, was isolated and determined to encode a polypeptide of 423 amino acids (46 kDa, pI = 6.53), present as a single copy in the I. fumosorosea genome. A split marker transformation system was developed and used to construct an Ifchit1 gene knockout. The ΔIfchit1 strain displayed minor alterations in mycelial growth on diverse media at 26 °C compared to the wild type and complemented (ΔIfchit1::Ifchit1) strains; however, colony morphology was affected, and the mutant strain had a temperature sensitive phenotype (32 °C). Although sporulation was delayed for the mutant, overall conidial production was almost twice than that of wild type. Biochemical assays indicated decreased chitinase activity during growth in Czapek-Dox liquid media for the ΔIfchit1 strain. Insect bioassays using diamondback moth, Plutella xylostella, larvae revealed decreased infectivity, i.e., increased LC 50 (threefold to fourfold) and a significantly delayed time to death, LT 50 from 3 to 6 days, for the ΔIfchit1 strain compared to the wild type and complemented strains. These data indicate an important role for the Ifchit1 chitinase as a virulence factor in I. fumosorosea. PMID:26910039

  7. Immunohistological localization of chitinase and -1,3-glucanase in rhizomania-diseased and benzothiadiazole treated sugar beet roots

    Czech Academy of Sciences Publication Activity Database

    Burketová, Lenka; Štillerová, Kateřina; Feltlová, Marcela

    2003-01-01

    Roč. 63, č. 1 (2003), s. 47-54. ISSN 0885-5765 R&D Projects: GA ČR GA522/03/0353 Institutional research plan: CEZ:AV0Z5038910 Keywords : immunolocalization * beta-1,3-glucanase * chitinase Subject RIV: CE - Biochemistry Impact factor: 1.262, year: 2003

  8. The Drosophila chitinase-like protein IDGF3 is involved in protection against nematodes and woung healing

    Czech Academy of Sciences Publication Activity Database

    Kučerová, Lucie; Brož, Václav; Arefin, B.; Maaroufi, H. O.; Hurychová, J.; Strnad, H.; Žurovec, Michal; Theopold, U.

    2016-01-01

    Roč. 8, č. 2 (2016), s. 199-210. ISSN 1662-811X R&D Projects: GA ČR GA14-27816S Institutional support: RVO:60077344 Keywords : chitinase-like proteins * imaginal disc growth factor * hemolymph clot Subject RIV: CE - Biochemistry Impact factor: 4.352, year: 2014 http://www.karger.com/Article/FullText/442351

  9. Induction by chromium ions of chitinases and polyamines in barley (Hordeum vulgare L.) and rape (Brassica napus L. ssp. oleifera)

    DEFF Research Database (Denmark)

    Jacobsen, S.; Hauschild, M.Z.; Rasmussen, U.

    1992-01-01

    Barley and rape seedlings were grown in hydroponic culture with increasing concentrations of CrO3 (Cr(VI)) or CrCl3 (Cr(III)). The chitinase activity and the concentrations of putrescine, spennidine and spermine were determined in the third leaf of barley seed-lings and in the second leaf of rape...

  10. Application of DNA bar codes for screening of industrially important fungi: the haplotype of Trichoderma harzianum sensu stricto indicates superior chitinase formation.

    Science.gov (United States)

    Nagy, Viviana; Seidl, Verena; Szakacs, George; Komoń-Zelazowska, Monika; Kubicek, Christian P; Druzhinina, Irina S

    2007-11-01

    Selection of suitable strains for biotechnological purposes is frequently a random process supported by high-throughput methods. Using chitinase production by Hypocrea lixii/Trichoderma harzianum as a model, we tested whether fungal strains with superior enzyme formation may be diagnosed by DNA bar codes. We analyzed sequences of two phylogenetic marker loci, internal transcribed spacer 1 (ITS1) and ITS2 of the rRNA-encoding gene cluster and the large intron of the elongation factor 1-alpha gene, tef1, from 50 isolates of H. lixii/T. harzianum, which were also tested to determine their ability to produce chitinases in solid-state fermentation (SSF). Statistically supported superior chitinase production was obtained for strains carrying one of the observed ITS1 and ITS2 and tef1 alleles corresponding to an allele of T. harzianum type strain CBS 226.95. A tef1-based DNA bar code tool, TrichoCHIT, for rapid identification of these strains was developed. The geographic origin of the strains was irrelevant for chitinase production. The improved chitinase production by strains containing this haplotype was not due to better growth on N-acetyl-beta-D-glucosamine or glucosamine. Isoenzyme electrophoresis showed that neither the isoenzyme profile of N-acetyl-beta-glucosaminidases or the endochitinases nor the intensity of staining of individual chitinase bands correlated with total chitinase in the culture filtrate. The superior chitinase producers did not exhibit similarly increased cellulase formation. Biolog Phenotype MicroArray analysis identified lack of N-acetyl-beta-D-mannosamine utilization as a specific trait of strains with the chitinase-overproducing haplotype. This observation was used to develop a plate screening assay for rapid microbiological identification of the strains. The data illustrate that desired industrial properties may be an attribute of certain populations within a species, and screening procedures should thus include a balanced mixture of all

  11. Oat (Avena sativa) seed extract as an antifungal food preservative through the catalytic activity of a highly abundant class I chitinase.

    Science.gov (United States)

    Sørensen, Hans Peter; Madsen, Lone Søvad; Petersen, Jørgen; Andersen, Jesper Tapdrup; Hansen, Anne Maria; Beck, Hans Christian

    2010-03-01

    Extracts from different higher plants were screened for the ability to inhibit the growth of Penicillium roqueforti, a major contaminating species in industrial food processing. Oat (Avena sativa) seed extracts exhibited a high degree of antifungal activity and could be used directly on rye bread to prevent the formation of P. roqueforti colonies. Proteins in the oat seed extracts were fractionated by column chromatography and proteins in fractions containing antifungal activity were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database searches. Identified antifungal candidates included thaumatin-like proteins, 1,3-beta-glucanase, permatin precursor, pathogenesis-related protein type 1, and chitinases of class I and II. Class I chitinase could be specifically removed from the extracts and was found to be indispensable for 50% of the P. roqueforti inhibiting activity. The purified class I chitinase has a molecular weight of approximately 34 kDa, optimal chitinase activity at pH 7, and exists as at least two basic isoforms (pI values of 7.6 and 8.0). Partial sequencing of the class I chitinase isoforms by LC-MS/MS revealed a primary structure with high similarity to class I chitinases of wheat (Triticum aestivum), barley (Hordeum vulgare), and rye (Secale cereale). Oat, wheat, barley, and rye seed extracts were compared with respect to the abundance of the class I chitinase and decrease in antifungal activity when class I chitinase is removed. We found that the oat seed class I chitinase is at least ten times more abundant than the wheat, barley, and rye homologs and that oat seed extracts are highly active toward P. roqueforti as opposed to extracts of other cereal seeds. PMID:19224400

  12. Quantitative Real-Time PCR Analysis of YKL-40 and Its Comparison with Mammalian Chitinase mRNAs in Normal Human Tissues Using a Single Standard DNA

    OpenAIRE

    Misa Ohno; Bauer, Peter O.; Yuta Kida; Masayoshi Sakaguchi; Yasusato Sugahara; Fumitaka Oyama

    2015-01-01

    YKL-40 (YKL for the first three N-terminal residues of a 40 kDa protein) belongs to a group of human chitinase-like proteins (CLPs), which are similar to chitinases but lack chitinolytic activity. YKL-40 mRNA and its protein levels have been reported elevated in multiple disorders including asthma, cystic fibrosis, rheumatoid arthritis and malignant tumors. Here, we quantified the YKL-40 mRNA levels and compared them with chitinases and housekeeping genes in normal human tissues. To establish...

  13. Chitinase production by Bacillus subtilis ATCC 11774 and its effect on biocontrol of Rhizoctonia diseases of potato.

    Science.gov (United States)

    Saber, Wesam I A; Ghoneem, Khalid M; Al-Askar, Abdulaziz A; Rashad, Younes M; Ali, Abeer A; Rashad, Ehsan M

    2015-12-01

    Stem canker and black scurf of potato, caused by Rhizoctonia solani, can be serious diseases causing an economically significant damage. Biocontrol activity of Bacillus subtilis ATCC 11774 against the Rhizoctonia diseases of potato was investigated in this study. Chitinase enzyme was optimally produced by B. subtilis under batch fermentation conditions similar to those of the potato-growing soil. The maximum chitinase was obtained at initial pH 8 and 30 °C. In vitro, the lytic action of the B. subtilis chitinase was detected releasing 355 μg GlcNAc ml⁻¹ from the cell wall extract of R. solani and suggesting the presence of various chitinase enzymes in the bacterial filtrate. In dual culture test, the antagonistic behavior of B. subtilis resulted in the inhibition of the radial growth of R. solani by 48.1% after 4 days. Moreover, the extracted B. subtilis chitinase reduced the growth of R. solani by 42.3% when incorporated with the PDA plates. Under greenhouse conditions, application of a bacterial suspension of B. subtilis at 109 cell mL⁻¹ significantly reduced the disease incidence of stem canker and black scurf to 22.3 and 30%, respectively. In addition, it significantly improved some biochemical parameters, growth and tubers yield. Our findings indicate two points; firstly, B. subtilis possesses a good biocontrol activity against Rhizoctonia diseases of potato, secondly, the harmonization and suitability of the soil conditions to the growth and activity of B. subtilis guaranteed a high controlling capacity against the target pathogen. PMID:26616375

  14. Insertion compounds of uranium oxides

    International Nuclear Information System (INIS)

    Insertion compounds are formed by the intercalation of an electropositive species, such as hydrogen or an alkali metal, with minimal structural rearrangement of the host oxide. In this report a review of the measured structural, thermodynamic and transport properties of the insertion compounds of α-U3O8, α-UO3, γ-UO3, δ-UO3 and related systems is given. (author)

  15. Transformation Fava Beans by Agrobacterium using Chitinase, Glucanase and CryIA (b genes

    Directory of Open Access Journals (Sweden)

    A.H Gorji

    2014-01-01

    Full Text Available In this study two plasmid vectors that are appropriate for plant transformation were made by preparation of gene cassettes for β-1, 3-glucanase from barley, chitinase from bean and cryIA (b from Bacillus thuringiensis (BT. Each of these genes were cloned under the control of the CaMV35S  promoter and the Nos terminator in pBI121 binary vector. pBI-Chi  and pBI-Glu and recombinant plasmid vectors were constructed via cloning of chitinase , β-1,3- glucanase and cryIA (b genes, respectively, instead of the gus gene in T-DNA region of pBI121 vector. Construction of pBI-ChiGlu recombinant plasmid vector was performed by means of cloning both of the complete chitinase and glucanase gene cassettes in pBI121 vector, with the intention of production synergistic effects against fungal infection.pBI-ChiBt  recombinant plasmid vector containing both of the complete chitinase and Bt gene cassettes was also constructed in order to contemporaneous plants resistance to pest pathogens and fungal in a single transformation event. pBI-ChiGlu and pBI-ChiBt that have been  introduced into the A. tumefaciens strain LBA4404 that was subsequently used for  transformation. Results indicate that embryogenic calli are well appropriate as objective material for Agrobacterium tumefaciens-mediated transformation in Faba bean.Seventeen well established shoots  were transferred to new MLS medium including suitable  antibiotics.Finally six independent transgenic plants were successfully rooted on kanamycin-containing  selection media and then transferred to soil after 20 days .Four plants out of six putative transgenic plants displaied to contain the end part of the chit transgene and nos terminator.The corresponding  piece, 700 bp of the chit gene, was amplified using specific primer.These putative transgenic plants were also be measured for the presence of the bgn13.1 and cryIA (b genes by PCR using specific primers.Two  pieces with expected sizes (1221 bp and 640

  16. [The accumulation of proteins with chitinase activity in the culture media of the parent and mutant Serratia marcescens strain grown in the presence of mitomycin C].

    Science.gov (United States)

    Iusupova, D V; Petukhova, E V; Sokolova, R B; Gabdrakhmanova, L A

    2002-01-01

    The study of the accumulation pattern of extracellular proteins with chitinase activity in the parent Serratia marcescens strain Bú 211 (ATCC 9986) grown in the presence of mitomycin C and its mutant strain with the constitutive synthesis of chitinases grown in the absence of the inducer showed that chitinase activity appeared in the culture liquids of both strains at the end of the exponential phase (4 h of growth) and reached a maximum in the stationary phase (18-20 h of growth). The analysis of the culture liquids (12 h of growth) by denaturing electrophoresis in PAAG followed by the protein renaturation step revealed the presence of four extracellular proteins with chitinase activity and molecular masses of 21, 38, 52, and 58 kDa. PMID:12449629

  17. Oat (Avena sativa) Seed Extract as an Antifungal Food Preservative Through the Catalytic Activity of a Highly Abundant Class I Chitinase

    DEFF Research Database (Denmark)

    Sørensen, Hans; Madsen, Lone; Petersen, Jørgen;

    2009-01-01

    Extracts from different higher plants were screened for the ability to inhibit the growth of Penicillium roqueforti, a major contaminating species in industrial food processing. Oat (Avena sativa) seed extracts exhibited a high degree of antifungal activity and could be used directly on rye bread...... similarity to class I chitinases of wheat (Triticum aestivum), barley (Hordeum vulgare), and rye (Secale cereale). Oat, wheat, barley, and rye seed extracts were compared with respect to the abundance of the class I chitinase and decrease in antifungal activity when class I chitinase is removed. We found...... that the oat seed class I chitinase is at least ten times more abundant than the wheat, barley, and rye homologs and that oat seed extracts are highly active toward P. roqueforti as opposed to extracts of other cereal seeds....

  18. Cerebrospinal fluid levels of chitinase 3-like 1 and neurofilament light chain predict multiple sclerosis development and disability after optic neuritis

    DEFF Research Database (Denmark)

    Modvig, S; Degn, M; Roed, H; Sørensen, Torben Lykke; Larsson, Hbw; Langkilde, Ar; Frederiksen, Jl; Sellebjerg, F

    evaluation. 18.6% were interviewed by phone. Cox regression, multiple regression and Spearman correlation analyses were used. RESULTS: Forty-six (53.5%) developed clinically definite MS (CDMS) during follow-up. In a multivariate model MRI (p=0.0001), chitinase 3-like 1 (p=0.0033) and age (p=0.0194) combined...... predicted CDMS best. Neurofilament light-chain predicted long-term disability by the multiple sclerosis severity scale (p=0.0111) and nine-hole-peg-test (p=0.0202). Chitinase-3-like-1 predicted long-term cognitive impairment by the paced auditory serial addition test (p=0.0150). CONCLUSION: Neurofilament...... light-chain and chitinase-3-like-1 were significant predictors of long-term physical and cognitive disability. Furthermore, chitinase-3-like-1 predicted CDMS development. Thus, these molecules hold promise as clinically valuable biomarkers after ON as a first demyelinating event....

  19. Domains and domain loss

    DEFF Research Database (Denmark)

    Haberland, Hartmut

    2005-01-01

    politicians and in the media, especially in the discussion whether some languages undergo ‘domain loss’ vis-à-vis powerful international languages like English. An objection that has been raised here is that domains, as originally conceived, are parameters of language choice and not properties of languages...... theoretical constructs that can explain language choice which were supposed to be a more powerful explanatory tool than more obvious (and observable) parameters like topic, place (setting) and interlocutor. In the meantime, at least in Scandinavia, the term ‘domain’ has been taken up in the debate among...

  20. Postharvest application of a novel chitinase cloned from Metschnikowia fructicola and overexpressed in Pichia pastoris to control brown rot of peaches.

    Science.gov (United States)

    Banani, Houda; Spadaro, Davide; Zhang, Dianpeng; Matic, Slavica; Garibaldi, Angelo; Gullino, Maria Lodovica

    2015-04-16

    Metschnikowia fructicola strain AP47 is a yeast antagonist against postharvest pathogens of fruits. The yeast was able to produce chitinase enzymes in the presence of pathogen cell wall. A novel chitinase gene MfChi (GenBank accession number HQ113461) was amplified from the genomic DNA of Metschnikowia fructicola AP47. Sequence analysis showed lack of introns, an open reading frame (ORF) of 1098 bp encoding a 365 amino acid protein with a calculated molecular weight of 40.9 kDa and a predicted pI of 5.27. MfChi was highly induced in Metschnikowia fructicola after interaction with Monilinia fructicola cell wall, suggesting a primary role of MfChi chitinase in the antagonistic activity of the yeast. The MfChi gene overexpressed in the heterologous expression system of Pichia pastoris KM71 and the recombinant chitinase showed high endochitinase activity towards 4-Nitrophenyl β-d-N,N',N″-triacetylchitotriose substrate. The antifungal activity of the recombinant chitinase was investigated against Monilinia fructicola and Monilinia laxa in vitro and on peaches. The chitinase significantly controlled the spore germination and the germ tube length of the tested pathogens in PDB medium and the mycelium diameter in PDA. The enzyme, when applied on peaches cv. Redhaven, successfully reduced brown rot severity. This work shows that the chitinase MfChi could be developed as a postharvest treatment with antimicrobial activity for fruit undergoing a short shelf life, and confirms that P. pastoris KM71 is a suitable microorganism for cost-effective large-scale production of recombinant chitinases. PMID:25632799

  1. Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifolia L.) show induction of chitinase activity upon mimicking the presence of prey

    OpenAIRE

    Matusikova, I.; Salaj, J.; Moravcikova, J.; Mlynarova, L.; Nap, J.P.H.; Libantova, J.

    2005-01-01

    Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addition, mechanical stirring of tentacles was performed. Chitinase activity was markedly increased in leaf exudates upon application of notably chitin. Application of gelatine increased the proteolyti...

  2. Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Its Application as Biocontrol for Whitefly (Bemisia tabaci Genn.)

    OpenAIRE

    NISA R MUBARIK; Irni Mahagiani; Amaryllis Anindyaputri; Sugeng Santoso; Iman Rusmana

    2010-01-01

    Problem statement: Chitin, a common constituent of insect exoskeleton, could be hydrolyzed by chitinase. The research was conducted to screen chitinolytic rhizobacteria isolated from rhizosphere of chilli pepper and to determine their chitinase activity in degrading chitin of whitefly, Bemisia tabaci Genn. (Hemiptera: Aleyrodidae). Whitefly is recognized as an important pest on many crops including chilli pepper. Approach: Screening and molecular identification based on 16...

  3. Gene Insertion Patterns and Sites

    Science.gov (United States)

    Vain, Philippe; Thole, Vera

    During the past 25 years, the molecular analysis of transgene insertion patterns and sites in plants has greatly contributed to our understanding of the mechanisms underlying transgene integration, expression, and stability in the nuclear genome. Molecular characterization is also an essential step in the safety assessment of genetically modified crops. This chapter describes the standard experimental procedures used to analyze transgene insertion patterns and loci in cereals and grasses transformed using Agrobacterium tumefaciens or direct transfer of DNA. Methods and protocols enabling the determination of the number and configuration of transgenic loci via a combination of inheritance studies, polymerase chain reaction, and Southern analyses are presented. The complete characterization of transgenic inserts in plants is, however, a holistic process relying on a wide variety of experimental approaches. In this chapter, these additional approaches are not detailed but references to relevant bibliographic records are provided.

  4. Partial purification, characterization, and kinetic studies of a low-molecular-weight, alkali-tolerant chitinase enzyme from Bacillus subtilis JN032305, A potential biocontrol strain.

    Science.gov (United States)

    Shivakumar, Srividya; Karmali, Anika Nayak; Ruhimbana, Charles

    2014-01-01

    A new alkalophilic low-molecular-mass chitinase of 14 kD from the potent biocontrol agent Bacillus subtilis JN032305 was partially purified and enzymology of the chitinase was studied. The enzyme showed optimal pH of 9.0 and temperature of 50°C. The enzyme was found stable during the 60-min incubation at 50 °C. The chitinase was inhibited by group specific agents like IAA, DAN, TLCK, and SDS and metal ions Mg(2+), Ca(2+), Fe(2+), Mn(2+), Ba(2+), and Hg(2+), whereas Zn(2+) did not show significant inhibitory effect against the chitinase. PMSF partially inhibited the enzyme. Substrates specificity tests indicated that the enzyme showed 75% of relative activity on glycol chitin, 58% on carboxymethylcellulose (CMC), 33% on chitin flakes, and 166% laminarin compared to that on colloidal chitin. The enzyme also hydrolyzed 4-methylumbelliferyl-N-acetyl-D-glucosaminide, indicating its chitobiase activity. The chitinase of this study has broad specificity, which could hydrolyze not only the glycosidic bond in GlcNAc-GlcNAc but also that of related carbohydrates with glycosidic linkages. The partially purified chitinase not only showed antifungal activity against Rhizoctonia solani and Colletotrichum gloeosporioides, two potent phytopathogens of chilli, but also increased the germination of chilli seeds when infected with the two potent phytopathogenic fungi. PMID:24499366

  5. Complete genome sequence of the fish pathogen Aeromonas veronii TH0426 with potential application in biosynthesis of pullulanase and chitinase.

    Science.gov (United States)

    Kang, Yuanhuan; Pan, Xiaoyi; Xu, Yang; Siddiqui, Shahrood A; Wang, Chunfeng; Shan, Xiaofeng; Qian, Aidong

    2016-06-10

    Aeromonas veronii TH0426 is a pathogen of the farmed yellow catfish Pelteobagrus fulvidraco but shows high-level expression of pullulanase and chitinase. Here, we present its genome sequence, which is the first reported complete genome of fish pathogen in A. veronii to date. Strain TH0426 harbors a single circular 4,923,009bp chromosome with a GC content of 58.25%. There are 4525 genes identified on its genome, including 4244 protein-coding genes, 32 rRNA genes, 120 tRNA genes, a noncoding RNA and 128 pseudo genes. We believe that the genomic information of A. veronii TH0426 would facilitate to reveal its pathogenic mechanism associated with yellow catfish, develop vaccine to decrease economic losses for fish farming, meanwhile explore the potential application in producing pullulanase and chitinase. PMID:27080448

  6. Concepts for stereoselective acrylate insertion

    KAUST Repository

    Neuwald, Boris

    2013-01-23

    Various phosphinesulfonato ligands and the corresponding palladium complexes [{((PaO)PdMeCl)-μ-M}n] ([{( X1-Cl)-μ-M}n], (PaO) = κ2- P,O-Ar2PC6H4SO2O) with symmetric (Ar = 2-MeOC6H4, 2-CF3C6H4, 2,6-(MeO)2C6H3, 2,6-(iPrO)2C 6H3, 2-(2′,6′-(MeO)2C 6H3)C6H4) and asymmetric substituted phosphorus atoms (Ar1 = 2,6-(MeO)2C6H 3, Ar2 = 2′-(2,6-(MeO)2C 6H3)C6H4; Ar1 = 2,6-(MeO)2C6H3, Ar2 = 2-cHexOC 6H4) were synthesized. Analyses of molecular motions and dynamics by variable temperature NMR studies and line shape analysis were performed for the free ligands and the complexes. The highest barriers of ΔGa = 44-64 kJ/mol were assigned to an aryl rotation process, and the flexibility of the ligand framework was found to be a key obstacle to a more effective stereocontrol. An increase of steric bulk at the aryl substituents raises the motional barriers but diminishes insertion rates and regioselectivity. The stereoselectivity of the first and the second methyl acrylate (MA) insertion into the Pd-Me bond of in situ generated complexes X1 was investigated by NMR and DFT methods. The substitution pattern of the ligand clearly affects the first MA insertion, resulting in a stereoselectivity of up to 6:1 for complexes with an asymmetric substituted phosphorus. In the consecutive insertion, the stereoselectivity is diminished in all cases. DFT analysis of the corresponding insertion transition states revealed that a selectivity for the first insertion with asymmetric (P aO) complexes is diminished in the consecutive insertions due to uncooperatively working enantiomorphic and chain end stereocontrol. From these observations, further concepts are developed. © 2012 American Chemical Society.

  7. Effect of phytoplasma infection on the activity of peroxidase, β-1,3 glucanase and chitinase in corn plants

    OpenAIRE

    Ana Carolina Bruno Junqueira; Ivan Paulo Bedendo; Sérgio Florentino Pascholati

    2011-01-01

    In the present work we studied the effect of inoculating corn plants with the maize bushy stunt phytoplasma on the activity of the enzymes peroxidase, β-1,3 glucanase and chitinase. The experiments were carried out inside a greenhouse. Plants of a resistant and a susceptible corn hybrid were inoculated by using infective Dalbulus maidis leafhoppers 10 days after sowing. When symptoms started to appear, leaf samples were collected at different periods to quantify enzyme activity. The results s...

  8. Turnabout Is Fair Play: Herbivory-Induced Plant Chitinases Excreted in Fall Armyworm Frass Suppress Herbivore Defenses in Maize.

    Science.gov (United States)

    Ray, Swayamjit; Alves, Patrick C M S; Ahmad, Imtiaz; Gaffoor, Iffa; Acevedo, Flor E; Peiffer, Michelle; Jin, Shan; Han, Yang; Shakeel, Samina; Felton, Gary W; Luthe, Dawn S

    2016-05-01

    The perception of herbivory by plants is known to be triggered by the deposition of insect-derived factors such as saliva and oral secretions, oviposition materials, and even feces. Such insect-derived materials harbor chemical cues that may elicit herbivore and/or pathogen-induced defenses in plants. Several insect-derived molecules that trigger herbivore-induced defenses in plants are known; however, insect-derived molecules suppressing them are largely unknown. In this study, we identified two plant chitinases from fall armyworm (Spodoptera frugiperda) larval frass that suppress herbivore defenses while simultaneously inducing pathogen defenses in maize (Zea mays). Fall armyworm larvae feed in enclosed whorls of maize plants, where frass accumulates over extended periods of time in close proximity to damaged leaf tissue. Our study shows that maize chitinases, Pr4 and Endochitinase A, are induced during herbivory and subsequently deposited on the host with the feces. These plant chitinases mediate the suppression of herbivore-induced defenses, thereby increasing the performance of the insect on the host. Pr4 and Endochitinase A also trigger the antagonistic pathogen defense pathway in maize and suppress fungal pathogen growth on maize leaves. Frass-induced suppression of herbivore defenses by deposition of the plant-derived chitinases Pr4 and Endochitinase A is a unique way an insect can co-opt the plant's defense proteins for its own benefit. It is also a phenomenon unlike the induction of herbivore defenses by insect oral secretions in most host-herbivore systems. PMID:26979328

  9. ECDYSTEROID AND CHITINASE FLUCTUATIONS IN THE WESTERN TARNISHED PLANT BUG (Lygus hesperus) PRIOR TO MOLT INDICATE ROLES IN DEVELOPMENT.

    Science.gov (United States)

    Brent, Colin S; Wang, Meixian; Miao, Yun-Gen; Hull, J Joe

    2016-06-01

    Vital physiological processes that drive the insect molt represent areas of interest for the development of alternative control strategies. The western tarnished plant bug (Lygus hesperus Knight) is a pest of numerous agronomic and horticultural crops but the development of novel control approaches is impeded by limited knowledge of the mechanisms regulating its molt. To address this deficiency, we examined the fundamental relationship underlying the hormonal and molecular components of ecdysis. At 27°C L. hesperus exhibits a temporally controlled nymph-adult molt that occurs about 4 days after the final nymph-nymph molt with ecdysteroid levels peaking 2 days prior to the final molt. Application of exogenous ecdysteroids when endogenous levels had decreased disrupted the nymphal-adult molt, with treated animals exhibiting an inability to escape the old exoskeleton and resulting in mortality compared to controls. Using accessible transcriptomic data, we identified 10 chitinase-like sequences (LhCht), eight of which had protein motifs consistent with chitinases. Phylogenetic analyses revealed orthologous relationships to chitinases critical to molting in other insects. RT-PCR based transcript profiling revealed that expression changes to four of the LhChts was coordinated with the molt period and ecdysteroid levels. Collectively, our results support a role for ecdysteroid regulation of the L. hesperus molt and suggest that cuticle clearance is mediated by LhCht orthologs of chitinases that are essential to the molt process. These results provide the initial hormonal and molecular basis for future studies to investigate the specific roles of these components in molting. PMID:27192063

  10. Purification of a thermostable chitinase from Bacillus cereus by chitin affinity and its application in microbial community changes in soil.

    Science.gov (United States)

    Liang, Tzu-Wen; Hsieh, Tung-Yen; Wang, San-Lang

    2014-06-01

    A thermostable chitinase was purified by chitin affinity from the culture supernatant of Bacillus cereus TKU028 with shrimp head powder (SHP) as the sole carbon/nitrogen source. TKU028 chitinase was purified using a one-step affinity adsorbent system, and the molecular mass of TKU028 chitinase (approximately 40 kDa) was then determined using SDS-PAGE. The enzyme was stable for 60 min at temperatures below 60 °C and stable over a broad pH range of 4-9 for 60 min. In addition, the temporal changes of a bacterial community in mangrove river sediment of the Tamsui River with added SHP were also analysed by PCR-denaturing gradient gel electrophoresis to investigate the effects of B. cereus TKU028 on the degradation of SHP. The 6-week incubation sample of SHP and B. cereus TKU028-amended mangrove river sediment displayed the highest amount of biomass, reducing sugar and total sugar, and some variance of bacterial community composition existed in the soils. PMID:24342954

  11. Characterization of a newly identified rice chitinase-like protein (OsCLP homologous to xylanase inhibitor

    Directory of Open Access Journals (Sweden)

    Wu Jingni

    2013-01-01

    Full Text Available Abstract Background During rice blast fungal attack, plant xylanase inhibitor proteins (XIPs that inhibit fungal xylanase activity are believed to act as a defensive barrier against fungal pathogens. To understand the role of XIPs in rice, a xylanase inhibitor was cloned from rice. The expression of this gene was examined at the transcriptional/translational levels during compatible and incompatible interactions, and the biochemical activity of this protein was also examined. Results Sequence alignment revealed that the deduced amino acid sequence of OsCLP shares a high degree of similarity with that of other plant TAXI-type XIPs. However, recombinant OsCLP did not display inhibitory activity against endo-1,4-β-xylanase enzymes from Aureobasidium pullulans (A. pullulans or Trichoderma viride (T. viride. Instead, an in-gel activity assay revealed strong chitinase activity. The transcription and translation of OsCLP were highly induced when rice was exposed to pathogens in an incompatible interaction. In addition, exogenous treatment with OsCLP affected the growth of the basidiomycete fungus Rhizoctonia solani through degradation of the hyphal cell wall. These data suggest that OsCLP, which has chitinase activity, may play an important role in plant defenses against pathogens. Conclusions Taken together, our results demonstrate that OsCLP may have antifungal activity. This protein may directly inhibit pathogen growth by degrading fungal cell wall components through chitinase activity.

  12. Comparative Evolutionary Histories of the Fungal Chitinase Gene Family Reveal Non-Random Size Expansions and Contractions due to Adaptive Natural Selection

    Directory of Open Access Journals (Sweden)

    Jan Stenlid

    2008-01-01

    Full Text Available Gene duplication and loss play an important role in the evolution of novel functions and for shaping an organism’s gene content. Recently, it was suggested that stress-related genes frequently are exposed to duplications and losses, while growth-related genes show selection against change in copy number. The fungal chitinase gene family constitutes an interesting case study of gene duplication and loss, as their biological roles include growth and development as well as more stress-responsive functions. We used genome sequence data to analyze the size of the chitinase gene family in different fungal taxa, which range from 1 in Batrachochytrium dendrobatidis and Schizosaccharomyces pombe to 20 in Hypocrea jecorina and Emericella nidulans, and to infer their phylogenetic relationships. Novel chitinase subgroups are identified and their phylogenetic relationships with previously known chitinases are discussed. We also employ a stochastic birth and death model to show that the fungal chitinase gene family indeed evolves non-randomly, and we identify six fungal lineages where larger-than-expected expansions (Pezizomycotina, H. jecorina, Gibberella zeae, Uncinocarpus reesii, E. nidulans and Rhizopus oryzae, and two contractions (Coccidioides immitis and S. pombe potentially indicate the action of adaptive natural selection. The results indicate that antagonistic fungal-fungal interactions are an important process for soil borne ascomycetes, but not for fungal species that are pathogenic in humans. Unicellular growth is correlated with a reduction of chitinase gene copy numbers which emphasizes the requirement of the combined action of several chitinases for filamentous growth.

  13. A small RNA controls expression of the chitinase ChiA in Listeria monocytogenes.

    Directory of Open Access Journals (Sweden)

    Jesper S Nielsen

    Full Text Available In recent years, more than 60 small RNAs (sRNAs have been identified in the gram-positive human pathogen Listeria monocytogenes, but their putative roles and mechanisms of action remain largely unknown. The sRNA LhrA was recently shown to be a post-transcriptional regulator of a single gene, lmo0850, which encodes a small protein of unknown function. LhrA controls the translation and degradation of the lmo0850 mRNA by an antisense mechanism, and it depends on the RNA chaperone Hfq for efficient binding to its target. In the present study, we sought to gain more insight into the functional role of LhrA in L. monocytogenes. To this end, we determined the effects of LhrA on global-wide gene expression. We observed that nearly 300 genes in L. monocytogenes are either positively or negatively affected by LhrA. Among these genes, we identified lmo0302 and chiA as direct targets of LhrA, thus establishing LhrA as a multiple target regulator. Lmo0302 encodes a hypothetical protein with no known function, whereas chiA encodes one of two chitinases present in L. monocytogenes. We show here that LhrA acts as a post-transcriptional regulator of lmo0302 and chiA by interfering with ribosome recruitment, and we provide evidence that both LhrA and Hfq act to down-regulate the expression of lmo0302 and chiA. Furthermore, in vitro binding experiments show that Hfq stimulates the base pairing of LhrA to chiA mRNA. Finally, we demonstrate that LhrA has a negative effect on the chitinolytic activity of L. monocytogenes. In marked contrast to this, we found that Hfq has a stimulating effect on the chitinolytic activity, suggesting that Hfq plays multiple roles in the complex regulatory pathways controlling the chitinases of L. monocytogenes.

  14. The first LHC insertion quadrupole

    CERN Multimedia

    2004-01-01

    An important milestone was reached in December 2003 at the CERN Magnet Assembly Facility. The team from the Accelerator Technology - Magnet and Electrical Systems group, AT-MEL, completed the first special superconducting quadrupole for the LHC insertions which house the experiments and major collider systems. The magnet is 8 metres long and contains two matching quadrupole magnets and an orbit corrector, a dipole magnet, used to correct errors in quadrupole alignment. All were tested in liquid helium and reached the ultimate performance criteria required for the LHC. After insertion in the cryostat, the superconducting magnet will be installed as the Q9 quadrupole in sector 7-8, the first sector of the LHC to be put in place in 2004. Members of the quadrupole team, from the AT-MEL group, gathered around the Q9 quadrupole at its inauguration on 12 December 2003 in building 181.

  15. Insertion device calculations with mathematica

    Energy Technology Data Exchange (ETDEWEB)

    Carr, R. [Stanford Synchrotron Radiation Lab., CA (United States); Lidia, S. [Univ. of California, Davis, CA (United States)

    1995-02-01

    The design of accelerator insertion devices such as wigglers and undulators has usually been aided by numerical modeling on digital computers, using code in high level languages like Fortran. In the present era, there are higher level programming environments like IDL{reg_sign}, MatLab{reg_sign}, and Mathematica{reg_sign} in which these calculations may be performed by writing much less code, and in which standard mathematical techniques are very easily used. The authors present a suite of standard insertion device modeling routines in Mathematica to illustrate the new techniques. These routines include a simple way to generate magnetic fields using blocks of CSEM materials, trajectory solutions from the Lorentz force equations for given magnetic fields, Bessel function calculations of radiation for wigglers and undulators and general radiation calculations for undulators.

  16. ATLAS insertable B-layer

    Czech Academy of Sciences Publication Activity Database

    Marčišovský, Michal

    2011-01-01

    Roč. 633, č. 1 (2011), "S224"-"S225". ISSN 0168-9002. [International workshop on radiation imaging detector s /11./. Praha, 26.06.2009-02.07.2009] R&D Projects: GA MŠk LA08015; GA MŠk LA08032 Institutional research plan: CEZ:AV0Z10100502 Keywords : ATLAS * pixel detector * insertable B-layer Subject RIV: BF - Elementary Particles and High Energy Physics Impact factor: 1.207, year: 2011

  17. Vacuum guidelines for ISA insertions

    International Nuclear Information System (INIS)

    Vacuum requirements place design restrictions on the ISA insertions. The vacuum tube diameter, given a distance L between pumps, is determined by the desorption of molecules from the wall under the impact of ions created by the beam, whereas the thickness of the tube must be sufficient to prevent collapse. In addition, the entire vacuum chamber must be able to be baked out at approximately 2000C

  18. HTS Insert Magnet Design Study

    CERN Document Server

    Devaux, M; Fleiter, J; Fazilleau, P; Lécrevisse, T; Pes, C; Rey, J-M; Rifflet, J-M; Sorbi, M; Stenvall, A; Tixador, P; Volpini, G

    2011-01-01

    Future accelerator magnets will need to reach higher field in the range of 20 T. This field level is very difficult to reach using only Low Temperature Superconductor materials whereas High Temperature Superconductors (HTS) provide interesting opportunities. High current densities and stress levels are needed to design such magnets. YBCO superconductor indeed carries large current densities under high magnetic field and provides good mechanical properties especially when produced using the IBAD approach. The HFM EUCARD program studies the design and the realization of an HTS insert of 6 T inside a Nb3Sn dipole of 13T at 4.2 K. In the2HTS insert, engineering current densities higher than 250 MA/m under 19 T are required to fulfill the specifications. The stress level is also very severe. YBCO IBAD tapes theoretically meet these challenges from presented measurements. The insert protection is also a critical because HTS materials show low quench propagation velocities and the coupling with the Nb3Sn magnet make...

  19. The Correlation between Chitin and Acidic Mammalian Chitinase in Animal Models of Allergic Asthma

    Directory of Open Access Journals (Sweden)

    Chia-Rui Shen

    2015-11-01

    Full Text Available Asthma is the result of chronic inflammation of the airways which subsequently results in airway hyper-responsiveness and airflow obstruction. It has been shown that an elicited expression of acidic mammalian chitinase (AMCase may be involved in the pathogenesis of asthma. Our recent study has demonstrated that the specific suppression of elevated AMCase leads to reduced eosinophilia and Th2-mediated immune responses in an ovalbumin (OVA-sensitized mouse model of allergic asthma. In the current study, we show that the elicited expression of AMCase in the lung tissues of both ovalbumin- and Der P2-induced allergic asthma mouse models. The effects of allergic mediated molecules on AMCase expression were evaluated by utilizing promoter assay in the lung cells. In fact, the exposure of chitin, a polymerized sugar and the fundamental component of the major allergen mite and several of the inflammatory mediators, showed significant enhancement on AMCase expression. Such obtained results contribute to the basis of developing a promising therapeutic strategy for asthma by silencing AMCase expression.

  20. Activity, stability and folding analysis of the chitinase from Entamoeba histolytica.

    Science.gov (United States)

    Muñoz, Patricia L A; Minchaca, Alexis Z; Mares, Rosa E; Ramos, Marco A

    2016-02-01

    Human amebiasis, caused by the parasitic protozoan Entamoeba histolytica, remains as a significant public health issue in developing countries. The life cycle of the parasite compromises two main stages, trophozoite and cyst, linked by two major events: encystation and excystation. Interestingly, the cyst stage has a chitin wall that helps the parasite to withstand harsh environmental conditions. Since the amebic chitinase, EhCHT1, has been recognized as a key player in both encystation and excystation, it is plausible to consider that specific inhibition could arrest the life cycle of the parasite and, thus, stop the infection. However, to selectively target EhCHT1 it is important to recognize its unique biochemical features to have the ability to control its cellular function. Hence, to gain further insights into the structure-function relationship, we conducted an experimental approach to examine the effects of pH, temperature, and denaturant concentration on the enzymatic activity and protein stability. Additionally, dependence on in vivo oxidative folding was further studied using a bacterial model. Our results attest the potential of EhCHT1 as a target for the design and development of new or improved anti-amebic therapeutics. Likewise, the potential of the oxidoreductase EhPDI, involved in oxidative folding of amebic proteins, was also confirmed. PMID:26526675

  1. Purification and characterization of three chitinases and one beta-1,3-glucanase accumulating in the medium of cell suspension cultures of barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Kragh, K.M.; Jacobsen, S.; Dalgaard Mikkelsen, J.;

    1991-01-01

    Three basic chitinases and one basic beta-1,3-glucanase were secreted into the medium when embryogenic cell suspensions of barley (Hordeum vulgare L.) cv. 'Igri' were cultured as undifferentiated aggregates in the presence of 2,4-D. The enzymes were purified by affinity and ion exchange......I at 8.7. Further analysis showed that this enzyme is also expressed in barley grain. The amino acid composition and five partial amino acid sequences covering 93 residues of chitinase K were determined. A high similarity was found between chitinase K and barley chitinase T and C as well as basic...... chitinases from barley aleurone and barley, bean and potato leaves. The purified beta-1,3-glucanase with a molecular weight (MW) of 32 kDa and pI greater-than-or-equal-to 9.8 constituted 1% of the soluble protein in the medium. Based on similar MW, pI and amino acid composition as well as identical N...

  2. The Chitinase Activity in Banana Seedling that Induce by Trichoderma spp as Resistance Responce to Fusarium Oxyporum f.sp.cubense

    Directory of Open Access Journals (Sweden)

    - Nurbailis

    2016-06-01

    Full Text Available An experiment was conducted to study the chitinase activities of banana seedling that induced by Trichoderma spp. The experiment consisted of two parts: 1. Testing chitinase activity in banana seedlings induced by Trichodernia spp., Using a factorial in a completely randomized design with two factors: a. types of inducers (biomass, liquid culture and filtrate, b. Isolates of Trichoderma spp. (T. Koningii-S6sh, T. Viride-T1sk, T.harzianum-P4sh and control. with 4 replications. Parameters measured were, the specific activity of the chitinase enzyme were detected in stems, leaves and roots of banana seedlings. 2. Testing of several types of inducers of Trichoderma spp. in inducing resistance in banana seedlings to Foc, together with the study design 1. Parameters measured were: incubation period, the percentage of symptomatic leaves and percentage discoloration of vessels. Result showed that 1. Liquid culture of induser  T. Viride -T1sk and T. koningii-S6sh and biomass of T. koningii-s6sh and T. harzianum-P4sh were effective in increasing  the specific activity of chitinase enzyme on banana seedling. The increase the amount specific activity of chitinase enzyme on banana seedling tissues by liquid culture of T. viride- T1sk 346% and 131,32% by T. koningii could be  suppress fusarium wilt desease on banana seedling. 

  3. Field Errors in Hybrid Insertion Devices

    OpenAIRE

    Schlueter, R.D.

    1995-01-01

    Hybrid magnet theory as applied to the error analyses used in the design of Advanced Light Source (ALS) insertion devices is reviewed. Sources of field errors in hybrid insertion devices are discussed.

  4. Field errors in hybrid insertion devices

    Energy Technology Data Exchange (ETDEWEB)

    Schlueter, R.D. [Lawrence Berkeley Lab., CA (United States)

    1995-02-01

    Hybrid magnet theory as applied to the error analyses used in the design of Advanced Light Source (ALS) insertion devices is reviewed. Sources of field errors in hybrid insertion devices are discussed.

  5. Delayed bowel perforation following suprapubic catheter insertion

    OpenAIRE

    Mehta Ajay; Ahmed Shwan J; Rimington Peter

    2004-01-01

    Abstract Background Complications of suprapubic catheter insertion are rare but can be significant. We describe an unusual complication of a delayed bowel perforation following suprapubic catheter insertion. Case presentation A gentleman presented with features of peritonitis and feculent discharge along a suprapubic catheter two months after insertion of the catheter. Conclusion Bowel perforation is the most feared complication of suprapubic catheter insertion especially in patients with low...

  6. Does 'insertion' work? France's minimum income"

    OpenAIRE

    Whitton, Timothy

    1993-01-01

    International audience France's Revenu Minimum d Insertion (RMI) received the approval of the French Parliament on 1st December1988, and came into effect shortly afterwardsi. In addition to a means test, the RMI is conditional upon signature of a contract (le contrat d'insertion) by which its recipients pledge themselves to take whatever action the RMI authorities recommend in order to re-insert themselves in main-stream society. Insertion is a difficult concept to translate into English. ...

  7. Regional insertion: an emergent approach

    International Nuclear Information System (INIS)

    The Brazilian Electrical Sector incorporates new variables that expressing the extensive spectrum of environmental impacts in the take of decisions, referring to the viability of realizing a electrical undertaking, attends the several restrictions that are important by the sector and by the society in the environment area and promotes the adequate generation of liquid benefits, consequential of the electrical undertaking. Due to these factors, the Electrical Sector is improving the concept of regional insertion, with the sectorial expansion in long-dated and the created demand in the environmental and social area, focalizing the solution for these questions. (C.G.C.). 1 fig, 2 tabs

  8. Tetrahedral mesh for needle insertion

    OpenAIRE

    Syvertsen, Rolf Anders

    2007-01-01

    This is a Master’s thesis in how to make a tetrahedral mesh for use in a needle insertion simulator. It also describes how it is possible to make the simulator, and how to improve it to make it as realistic as possible. The medical simulator uses a haptic device, a haptic scene graph and a FEM for realistic soft tissue deformation and interaction. In this project a tetrahedral mesh is created from a polygon model, and then the mesh has been loaded into the HaptX haptic scene graph. The object...

  9. Domain Engineering

    Science.gov (United States)

    Bjørner, Dines

    Before software can be designed we must know its requirements. Before requirements can be expressed we must understand the domain. So it follows, from our dogma, that we must first establish precise descriptions of domains; then, from such descriptions, “derive” at least domain and interface requirements; and from those and machine requirements design the software, or, more generally, the computing systems.

  10. Chitinase-like proteins promote IL-17-mediated neutrophilia in a trade-off between nematode killing and host damage

    OpenAIRE

    Sutherland, Tara E.; Logan, Nicola; Rückerl, Dominik; Alison A Humbles; Allan, Stuart M.; Papayannopoulos, Venizelos; Stockinger, Brigitta; Maizels, Rick M.; Allen, Judith E.

    2014-01-01

    Enzymatically inactive chitinase-like proteins (CLPs) such as BRP-39, Ym1 and Ym2 are established markers of immune activation and pathology, yet their functions are essentially unknown. We show that Ym1 and Ym2 induce neutrophil accumulation through expansion of interleukin 17 (IL-17)-producing γδ T cells. While BRP-39 did not influence neutrophilia, it was required for IL-17 production in γδ T cells, suggesting IL-17 regulation is an inherent feature of murine CLPs. Using a model of lung mi...

  11. Characterization of a novel Salmonella typhimurium chitinase which hydrolyzes chitin, chitooligosaccharides and an N-acetyllactosamine conjugate

    DEFF Research Database (Denmark)

    Larsen, Tanja; Petersen, Bent O.; Storgaard, Birgit Groth;

    2011-01-01

    Salmonella contain genes annotated as chitinases; however, their chitinolytic activities have never been verified. We now demonstrate such an activity for a chitinase assigned to glycoside hydrolase family 18 encoded by the SL0018 (chiA) gene in Salmonella enterica Typhimurium SL1344. A C...... carboxymethyl chitin Remazol Brilliant Violet but does not act on 4-nitrophenyl N-acetyl-ß-D-glucosaminide, peptidoglycan or 4-nitrophenyl ß-D-cellobioside. Enzyme activity was also characterized by directly monitoring product formation using (1)H-nuclear magnetic resonance which showed that chitin is a...

  12. Effect of Botrytis cinerea infection and elicitation on ß-1,3-glucanase and chitinase activity in bean leaves and cell cultures

    Directory of Open Access Journals (Sweden)

    Elżbieta Kuźniak

    2013-12-01

    Full Text Available The activity of ß-1,3-glucanase and chitinase in bean plants treated with B. cinerea products or/and infected and in cell cultures after application of fungal products has been studied. Botrytis cinerea infection and culture filtrates, ethanol precipitates, glucan and conidial extract treatment markedly enhanced the activity of both hydrolases. Cell cultures treated with B.cinerea products reacted similarly to intact plants. In plants pretreated with 2-day culture filtrate and conidial extract and then infected, ß-1,3-glucanase and chitinase were induced stronger than after infection without pretreatment.

  13. Flexibility in targeting and insertion during bacterial membrane protein biogenesis

    International Nuclear Information System (INIS)

    The biogenesis of Escherichia coli inner membrane proteins (IMPs) is assisted by targeting and insertion factors such as the signal recognition particle (SRP), the Sec-translocon and YidC with translocation of (large) periplasmic domains energized by SecA and the proton motive force (pmf). The use of these factors and forces is probably primarily determined by specific structural features of an IMP. To analyze these features we have engineered a set of model IMPs based on endogenous E. coli IMPs known to follow distinct targeting and insertion pathways. The modified model IMPs were analyzed for altered routing using an in vivo protease mapping approach. The data suggest a facultative use of different combinations of factors

  14. 华山松疱锈病的重寄生真菌(深绿木霉)中几丁质酶基因cDNA片段的克隆%Cloning of cDNA Fragment of Chitinase Gene from the Mycoparasite Trichoderma atroviride on Armandii Pine Blister Rust

    Institute of Scientific and Technical Information of China (English)

    马长乐; 李靖; 陈玉惠; 刘小烛

    2008-01-01

    [Objective] The aim of this study was to isolate chitinase gene from Trichoderma atroviride strain SS003. [Method] With the aeciospore wall of armandii pine blister rust as inducer, chitinase gene was induced to express in Trichoderma atroviride cells. The cDNA fragment of chitinase gene was cloned by RT-PCR approach. [Result] The activity of chitinase induced reached 40.17 μg/10 min; and the specific fragment amplified was 834 bp in length and proved to be the fragment of chitinase gene by sequencing and sequence analysis. [Conclusion] The result showed the feasibility of isolating the full length of chitinase gene and its transformation, and further producing chitinase.

  15. Heterogonous expression and characterization of a plant class IV chitinase from the pitcher of the carnivorous plant Nepenthes alata.

    Science.gov (United States)

    Ishisaki, Kana; Honda, Yuji; Taniguchi, Hajime; Hatano, Naoya; Hamada, Tatsuro

    2012-03-01

    A class IV chitinase belonging to the glycoside hydrolase 19 family from Nepenthes alata (NaCHIT1) was expressed in Escherichia coli. The enzyme exhibited weak activity toward polymeric substrates and significant activity toward (GlcNAc)(n) [β-1,4-linked oligosaccharide of GlcNAc with a polymerization degree of n (n = 4-6)]. The enzyme hydrolyzed the third and fourth glycosidic linkages from the non-reducing end of (GlcNAc)(6). The pH optimum of the enzymatic reaction was 5.5 at 37°C. The optimal temperature for activity was 60°C in 50 mM sodium acetate buffer (pH 5.5). The anomeric form of the products indicated that it was an inverting enzyme. The k(cat)/K(m) of the (GlcNAc)(n) hydrolysis increased with an increase in the degree of polymerization. Amino acid sequence alignment analysis between NaCHIT1 and a class IV chitinase from a Picea abies (Norway spruce) suggested that the deletion of four loops likely led the enzyme to optimize the (GlcNAc)(n) hydrolytic reaction rather than the hydrolysis of polymeric substrates. PMID:21930651

  16. PanoInserts: mobile spatial teleconferencing

    OpenAIRE

    Pece, F.; Steptoe, W.; F. Wanner; Julier, S.; Weyrich, T.; Kautz, J.; Steed, A.

    2013-01-01

    We present PanoInserts: a novel teleconferencing system that uses smartphone cameras to create a surround representation of meeting places. We take a static panoramic image of a location into which we insert live videos from smartphones. We use a combination of marker- and image-based tracking to position the video inserts within the panorama, and transmit this representation to a remote viewer. We conduct a user study comparing our system with fully-panoramic video and conventional webcam vi...

  17. Delayed bowel perforation following suprapubic catheter insertion

    Directory of Open Access Journals (Sweden)

    Mehta Ajay

    2004-12-01

    Full Text Available Abstract Background Complications of suprapubic catheter insertion are rare but can be significant. We describe an unusual complication of a delayed bowel perforation following suprapubic catheter insertion. Case presentation A gentleman presented with features of peritonitis and feculent discharge along a suprapubic catheter two months after insertion of the catheter. Conclusion Bowel perforation is the most feared complication of suprapubic catheter insertion especially in patients with lower abdominal scar. The risk may be reduced with the use of ultrasound scan guidance.

  18. Nozzle insert for mixed mode fuel injector

    Science.gov (United States)

    Lawrence, Keith E.

    2006-11-21

    A fuel injector includes a homogenous charge nozzle outlet set and a conventional nozzle outlet set controlled respectively, by first and second needle valve members. The homogeneous charged nozzle outlet set is defined by a nozzle insert that is attached to an injector body, which defines the conventional nozzle outlet set. The nozzle insert is a one piece metallic component with a large diameter segment separated from a small diameter segment by an annular engagement surface. One of the needle valve members is guided on an outer surface of the nozzle insert, and the nozzle insert has an interference fit attachment to the injector body.

  19. A HTS dipole insert coil constructed

    CERN Document Server

    Ballarino, A; Rey, J M; Stenvall, A; Sorbi, M; Tixador, P

    2013-01-01

    This report is the deliverable report 7.4.1 “A HTS dipole insert coil constructed“. The report has three parts: “Design report for the HTS dipole insert”, “One insert pancake prototype coil constructed with the setup for a high field test”, and “All insert components ordered”. The three report parts show that, although the insert construction will be only completed by end 2013, all elements are present for a successful completion and that, given the important investments done by the participants, there is a full commitment of all of them to finish the project

  20. Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifolia L.) show induction of chitinase activity upon mimicking the presence of prey

    NARCIS (Netherlands)

    Matusikova, I.; Salaj, J.; Moravcikova, J.; Mlynarova, L.; Nap, J.P.H.; Libantova, J.

    2005-01-01

    Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addi

  1. Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifoliaL.) show induction of chitinase activity upon mimicking the presence of prey

    NARCIS (Netherlands)

    Matusikova, [No Value; Salaj, J; Moravcikova, J; Mlynarova, L; Nap, JP; Libantova, J

    2005-01-01

    Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addi

  2. Introduction of a tryptophan side chain into subsite +1 enhances transglycosylation activity of a GH-18 chitinase from Arabidopsis thaliana, AtChiC

    DEFF Research Database (Denmark)

    Umemoto, Naoyuki; Ohnuma, Takayuki; Mizuhara, Mamiko; Sato, Hirokazu; Skriver, Karen; Fukamizo, Tamo

    2013-01-01

    A tryptophan side chain was introduced into subsite +1 of family GH-18 (class V) chitinases from Nicotiana tabacum and Arabidopsis thaliana (NtChiV and AtChiC, respectively) by the mutation of a glycine residue to tryptophan (G74W-NtChiV and G75W-AtChiC). The specific activity toward glycol chitin...

  3. Multipurpose Transposon-Insertion Libraries in Yeast.

    Science.gov (United States)

    Kumar, Anuj

    2016-01-01

    Libraries of transposon-insertion alleles constitute powerful and versatile tools for large-scale analysis of yeast gene function. Transposon-insertion libraries are constructed most simply through mutagenesis of a plasmid-based genomic DNA library; modification of the mutagenizing transposon by incorporation of yeast selectable markers, recombination sites, and an epitope tag enables the application of insertion alleles for phenotypic screening and protein localization. In particular, yeast genomic DNA libraries have been mutagenized with modified bacterial transposons carrying the URA3 marker, lox recombination sites, and sequence encoding multiple copies of the hemagglutinin (HA) epitope. Mutagenesis with these transposons has yielded a large resource of insertion alleles affecting nearly 4000 yeast genes in total. Through well-established protocols, these insertion libraries can be introduced into the desired strain backgrounds and the resulting insertional mutants can be screened or systematically analyzed. Relative to alternative methods of UV irradiation or chemical mutagenesis, transposon-insertion alleles can be easily identified by PCR-based approaches or high-throughput sequencing. Transposon-insertion libraries also provide a cost-effective alternative to targeted deletion approaches, although, in contrast to start-codon to stop-codon deletions, insertion alleles might not represent true null-mutants. For protein-localization studies, transposon-insertion alleles can provide encoded epitope tags in-frame with internal codons; in many cases, these transposon-encoded epitope tags can provide a more accurate localization for proteins in which terminal sequences are crucial for intracellular targeting. Thus, overall, transposon-insertion libraries can be used quickly and economically and have a particular utility in screening for desired phenotypes and localization patterns in nonstandard genetic backgrounds. PMID:27250950

  4. Quantitative Real-Time PCR Analysis of YKL-40 and Its Comparison with Mammalian Chitinase mRNAs in Normal Human Tissues Using a Single Standard DNA

    Directory of Open Access Journals (Sweden)

    Misa Ohno

    2015-04-01

    Full Text Available YKL-40 (YKL for the first three N-terminal residues of a 40 kDa protein belongs to a group of human chitinase-like proteins (CLPs, which are similar to chitinases but lack chitinolytic activity. YKL-40 mRNA and its protein levels have been reported elevated in multiple disorders including asthma, cystic fibrosis, rheumatoid arthritis and malignant tumors. Here, we quantified the YKL-40 mRNA levels and compared them with chitinases and housekeeping genes in normal human tissues. To establish the quantitative real-time PCR (qPCR system for evaluation of relative YKL-40 mRNA levels, we constructed a human standard DNA molecule by ligating cDNAs of YKL-40, two mammalian chitinases and two housekeeping genes in a one-to-one ratio. We generated cDNAs from various normal human tissues and analyzed the YKL-40 mRNA expression levels using a qPCR system with the standard DNA. We found that YKL-40 mRNA is present widely in human tissues while its expression patterns exhibit clear tissue specificity. Highest YKL-40 mRNA levels were detected in the liver, followed by kidney, trachea and lung. The levels of YKL-40 mRNA in the kidney and liver were more than 100-times higher than those of chitotriosidase mRNA. Our study provides for the first time a comprehensive analysis of the relative expression levels of YKL-40 mRNA versus mammalian chitinases in normal human tissues.

  5. Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain.

    OpenAIRE

    Patel, N; Thierry-Mieg, D.; Mancillas, J R

    1993-01-01

    An additional genetic locus in Caenorhabditis elegans, unc-116, was identified in a screen for mutations resulting in defective locomotion. unc-116 was cloned by use of a transposon insertion mutant and the physical and genetic map of the genome. The cDNA sequence predicts an 815-amino acid protein. Based upon sequence comparison and secondary structure predictions, unc-116 encodes all three domains of the kinesin heavy chain: the motor, stalk, and tail. While the motor and tail domains have ...

  6. Central Solenoid Insert Technical Specification

    Energy Technology Data Exchange (ETDEWEB)

    Martovetsky, Nicolai N [ORNL; Smirnov, Alexandre [ORNL

    2011-09-01

    The US ITER Project Office (USIPO) is responsible for the ITER central solenoid (CS) contribution to the ITER project. The Central Solenoid Insert (CSI) project will allow ITER validation the appropriate lengths of the conductors to be used in the full-scale CS coils under relevant conditions. The ITER Program plans to build and test a CSI to verify the performance of the CS conductor. The CSI is a one-layer solenoid with an inner diameter of 1.48 m and a height of 4.45 m between electric terminal ends. The coil weight with the terminals is approximately 820 kg without insulation. The major goal of the CSI is to measure the temperature margin of the CS under the ITER direct current (DC) operating conditions, including determining sensitivity to load cycles. Performance of the joints, ramp rate sensitivity, and stability against thermal or electromagnetic disturbances, electrical insulation, losses, and instrumentation are addressed separately and therefore are not major goals in this project. However, losses and joint performance will be tested during the CSI testing campaign. The USIPO will build the CSI that will be tested at the Central Solenoid Model Coil (CSMC) Test Facility at the Japan Atomic Energy Agency (JAEA), Naka, Japan. The industrial vendors (the Suppliers) will report to the USIPO (the Company). All approvals to proceed will be issued by the Company, which in some cases, as specified in this document, will also require the approval of the ITER Organization. Responsibilities and obligations will be covered by respective contracts between the USIPO, called Company interchangeably, and the industrial Prime Contractors, called Suppliers. Different stages of work may be performed by more than one Prime Contractor, as described in this specification. Technical requirements of the contract between the Company and the Prime Contractor will be covered by the Fabrication Specifications developed by the Prime Contractor based on this document and approved by

  7. A fuel insert shuffle planner expert system

    International Nuclear Information System (INIS)

    During the refueling outage of a pressurized water reactor, the fuel assembly inserts must be moved from their current fuel assemblies to the proper fuel assemblies for the next cycle core design. The crane movement pattern, or shuffle plan, to perform this operation should be as efficient as possible because a less efficient plan could cause the power plant to be shut down for a longer period of time. The current method to develop this plan requires three to four man-weeks of a reactor engineer's time. This plan is developed by the reactor engineer who uses his expertise from previous insert shuffles. Further modifications to this plan are sometimes required due to either changes in the desired core load pattern or damaged fuel or inserts. These changes generally require two to four man-days during the plant outage time. A full-scale prototype of an insert shuffle advisor expert system has been developed to create the insert shuffle plan. This system substantially reduces the manpower requirements needed to develop this plan. It is also a vehicle for preserving the expertise needed to develop this plan. Since the plan can be developed more quickly with the insert shuffle advisor, several strategies can be tried. This gives the potential for a more efficient insert shuffle plan than could be found by the reactor engineer. A more efficient plan means that the power plant could be shut down for a shorter period of time. This would have considerable financial advantages

  8. Identification and Antagonism Study of a Novel Chitinase-producing Bacterium Burkholderia Sp.C3 against Phytopathogenic Fungi

    Institute of Scientific and Technical Information of China (English)

    金虹; TAO Yong

    2006-01-01

    Through a modified agar well diffusion assay, antagonism of a novel chitinase-producing strain C3 against the phytopathogenic fungi including Phoma wasabiae Yokogi,Heterostrophus, Exserohilum Turcicum, Curwularia (Walk) Boed, Thantephorus cucumris, Fusarium graminearum was tested. The data showed that the crude cxtracts of strain C3 had stable antifungal activity in the range of pH 5.0 to pH 8.0. The active components were heat labile and sensitive to proteinase K. A series of experiments supported that the compound responsible for inhibitory activity appeared to be ehitinase. The 16s rDNA analysis indicated that C3 was subject to genus Burkholderia. Pbenotypic characterization of C3 was also consisted with the result of molecular identification.

  9. The chitinase-like protein YKL-40 increases mucin5AC production in human bronchial epithelial cells

    International Nuclear Information System (INIS)

    Mucus overproduction is an important feature in patients with chronic inflammatory airway diseases. However, the regulatory mechanisms that mediate excessive mucin production remain elusive. Recently, the level of YKL-40, a chitinase-like protein, has been found to be significantly increased in chronic inflammatory airway diseases and has been shown to be associated with the severity of these diseases. In this study, we sought to explore the effect of YKL-40 on mucin5AC (MUC5AC) production in chronic inflammatory airway diseases and the potential signaling pathways involved in this process. We found that elevated YKL-40 levels increased the mRNA and protein expression of MUC5AC in a dose- and time-dependent manner, in association with the phosphorylation of extracellular signal-regulated kinase (ERK) and nuclear factor κB (NF-κB), reflecting their activation. These responses were significantly suppressed by the knockdown of protease-activating receptor 2 (PAR2) with specific small interfering RNA or the inhibitors of ERK and NF-κB. YKL-40-induced MUC5AC overproduction was also effectively attenuated by the inhibitor of focal adhesion kinase (FAK). Taken together, these results imply that YKL-40 can stimulate excessive MUC5AC production through PAR2- and FAK-mediated mechanisms. - Highlights: • MUC5AC is the major secreted mucin in chronic inflammatory airway diseases. • YKL-40 is a prototype of the chitinase-like protein in mammals. • YKL-40 is an active player in chronic inflammatory airway diseases. • YKL-40 can increase MUC5AC production via PAR2-mediated pathway. • FAK is another candidate to mediate YKL-40-induced MUC5AC overexpression

  10. Transgenic Indian Cotton (Gossypium hirsutum Harboring Rice Chitinase Gene (Chi II Confers Resistance to Two Fungal Pathogens

    Directory of Open Access Journals (Sweden)

    M. Ganesan

    2009-01-01

    Full Text Available Problem statement: The present investigation described a simple and reproducible protocol for transgenic cotton regeneration and characterization of chitinase (Chi II gene expression against two different fungal pathogens in cotton. Approach: Transgenic cotton (Gossypium hirsutum cv. SVPR2 plants were produced by pCambia-bar-Chi II (13.8 kb under the control of the CaMV 35S promoter, harbored in the strain LBA 4404 Agrobacterium tumefaciens by using shoot tip explants. Results: Finally, from the 10 experiments, 21.8% of transformation frequency was recorded. Segregation ratio of 3:1 was recorded in the T0 plant seeds. Polymerase chain reaction and southern blotting analysis were used to confirm the integration of Chi II transgene in the T0 plants genome of putative transgenics. Quantitiave and qualitative (SDS-PAGE analyses were also carried out to confirm the expression of chitinase enzyme in T0 plants. Further, randomly selected transgenic plants (T0 were analyzed for disease tolerance by evaluating them with spores of Fusarium oxysporum and Alternaria macrospora. All the selected PCR positive plants showed enhanced disease resistance against Fusarium wilt. The plants selected randomly showed an enhanced survival rate compared with the control when they were grown in earthen pots inoculated with 1×105 spores 100-1 g of soil mixture. Another four randomly selected plantlets were sprayed with spores of Alternaria macrospora in order to test their tolerance to Alternaria leaf spot disease. After 20 days of culture, the number of lesions per leaf and the lesion length per leaf spot in non-transferred leaves increased. In the case of transgenic plantlets, lesion formation was completely absent. Conclusion: The disease resistance against Fusarium wilt and Alternaria leaf spot in cotton strains would serve as good breeding materials for producing fungal disease resistant cotton varieties.

  11. The chitinase-like protein YKL-40 increases mucin5AC production in human bronchial epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chunyi; Li, Qi [Division of Respiratory Medicine, Second Affiliated Hospital, Chongqing Medical University, No. 74, Linjiang Road, Yuzhong District, Chongqing 400010 (China); Zhou, Xiangdong, E-mail: zxd999@263.net [Division of Respiratory Medicine, Second Affiliated Hospital, Chongqing Medical University, No. 74, Linjiang Road, Yuzhong District, Chongqing 400010 (China); Kolosov, Victor P.; Perelman, Juliy M. [Far Eastern Scientific Center of Physiology and Pathology of Respiration, Siberian Branch, Russian Academy of Medical Sciences, Blagoveshchensk (Russian Federation)

    2013-11-01

    Mucus overproduction is an important feature in patients with chronic inflammatory airway diseases. However, the regulatory mechanisms that mediate excessive mucin production remain elusive. Recently, the level of YKL-40, a chitinase-like protein, has been found to be significantly increased in chronic inflammatory airway diseases and has been shown to be associated with the severity of these diseases. In this study, we sought to explore the effect of YKL-40 on mucin5AC (MUC5AC) production in chronic inflammatory airway diseases and the potential signaling pathways involved in this process. We found that elevated YKL-40 levels increased the mRNA and protein expression of MUC5AC in a dose- and time-dependent manner, in association with the phosphorylation of extracellular signal-regulated kinase (ERK) and nuclear factor κB (NF-κB), reflecting their activation. These responses were significantly suppressed by the knockdown of protease-activating receptor 2 (PAR2) with specific small interfering RNA or the inhibitors of ERK and NF-κB. YKL-40-induced MUC5AC overproduction was also effectively attenuated by the inhibitor of focal adhesion kinase (FAK). Taken together, these results imply that YKL-40 can stimulate excessive MUC5AC production through PAR2- and FAK-mediated mechanisms. - Highlights: • MUC5AC is the major secreted mucin in chronic inflammatory airway diseases. • YKL-40 is a prototype of the chitinase-like protein in mammals. • YKL-40 is an active player in chronic inflammatory airway diseases. • YKL-40 can increase MUC5AC production via PAR2-mediated pathway. • FAK is another candidate to mediate YKL-40-induced MUC5AC overexpression.

  12. Evaluation of kinetic parameters of chitinases produced by Beauveria bassiana (Bals.) Vuill. /
    Avaliação de parâmetros cinéticos de quitinases produzidas por Beauveria bassiana (Bals.) Vuill.

    OpenAIRE

    Cristiane Mita; Vanessa Hitomi Sugahara; Jo I Wu; Pedro Manoel Oliveira Janeiro Neves; Dalva Tomoe Miyagui; Geni Varéa-Pereira; Danieli Cristina Sassá; Evelyn Kamogawa

    2008-01-01

    Entomopathogenic fungus Beauveria bassiana is currently used as a biocontrol agent for agricultural pests. The infection process involves extracellular enzymes such as proteases and chitinases that degrade the cuticle of the insects. The objective of this work was to evaluate kinetic parameters of pH, temperature, ionic concentration and time of reaction on chitinases activity. The fungus B. bassiana CG432 was cultivated on coffee berry borer Hypothenemus hampei (Ferrari) and the conidia grow...

  13. HB+ inserted into the CMS Solenoid

    CERN Multimedia

    Tejinder S. Virdee, CERN

    2006-01-01

    The first half of the barrel hadron calorimeter (HB+) has been inserted into the superconducting solenoid of CMS, in preparation for the magnet test and cosmic challenge. The operation went smoothly, lasting a couple of days.

  14. An Unusual Complication of Suprapubic Catheter Insertion

    Directory of Open Access Journals (Sweden)

    Krishnan Ananthakrishnan

    2006-01-01

    Full Text Available A patient who had a small bowel mesentery perforation following insertion of a suprapubic catheter (SPC is described. He had no bowel complaints immediately following the procedure, but presented 10 weeks later with insidious onset bowel obstruction due to the kink caused by the catheter. This complication occurred despite cystoscopy control and adequate bladder distension prior to the procedure. This isolated case illustrates the fact that regardless of the ease and frequency of SPC insertion, complications do occur.

  15. Strength of inserts in titanium alloy machining

    Science.gov (United States)

    Kozlov, V.; Huang, Z.; Zhang, J.

    2016-04-01

    In this paper, a stressed state of a non-worn cutting wedge in a machined titanium alloy (Ti6Al2Mo2Cr) is analyzed. The distribution of contact loads on the face of a cutting tool was obtained experimentally with the use of a ‘split cutting tool’. Calculation of internal stresses in the indexable insert made from cemented carbide (WC8Co) was carried out with the help of ANSYS 14.0 software. Investigations showed that a small thickness of the cutting insert leads to extremely high compressive stresses near the cutting edge, stresses that exceed the ultimate compressive strength of cemented carbide. The face and the base of the insert experience high tensile stresses, which approach the ultimate tensile strength of cemented carbide and increase a probability of cutting insert destruction. If the thickness of the cutting insert is bigger than 5 mm, compressive stresses near the cutting edge decrease, and tensile stresses on the face and base decrease to zero. The dependences of the greatest normal and tangential stresses on thickness of the cutting insert were found. Abbreviation and symbols: m/s - meter per second (cutting speed v); mm/r - millimeter per revolution (feed rate f); MPa - mega Pascal (dimension of specific contact loads and stresses); γ - rake angle of the cutting tool [°] α - clearance angle of the sharp cutting tool [°].

  16. Z-2 Threaded Insert Design and Testing

    Science.gov (United States)

    Ross, Amy; Rhodes, Richard; Jones, Robert J.; Graziosi, David; Ferl, Jinny; Sweeny, Mitch; Scarborough, Stephen

    2016-01-01

    NASA's Z-2 prototype space suit contains several components fabricated from an advanced hybrid composite laminate consisting of IM10 carbon fiber and fiber glass. One requirement was to have removable, replaceable helicoil inserts to which other suit components would be fastened. An approach utilizing bonded in inserts with helicoils inside of them was implemented. During initial assembly, cracking sounds were heard followed by the lifting of one of the blind inserts out of its hole when the screws were torqued. A failure investigation was initiated to understand the mechanism of the failure. Ultimately, it was determined that the pre-tension caused by torqueing the fasteners is a much larger force than induced from the pressure loads of the suit which was not considered in the insert design. Bolt tension is determined by dividing the torque on the screw by a k value multiplied by the thread diameter of the bolt. The k value is a factor that accounts for friction in the system. A common value used for k for a non-lubricated screw is 0.2. The k value can go down by as much as 0.1 if the screw is lubricated which means for the same torque, a much larger tension could be placed on the bolt and insert. This paper summarizes the failure investigation that was performed to identify the root cause of the suit failure and details how the insert design was modified to resist a higher pull out tension.

  17. Domain crossing

    DEFF Research Database (Denmark)

    Schraefel, M. C.; Rouncefield, Mark; Kellogg, Wendy; Ackerman, Mark; Marsden, Gary; Bødker, Susanne; Wyche, Susan; Reddy, Madhu

    In CSCW, how much do we need to know about another domain/culture before we observe, intersect and intervene with designs. What optimally would that other culture need to know about us? Is this a “how long is a piece of string” question, or an inquiry where we can consider a variety of contexts a...

  18. Recurrent insertion of 5′-terminal nucleotides and loss of the branchpoint motif in lineages of group II introns inserted in mitochondrial preribosomal RNAs

    OpenAIRE

    Li, Cheng-Fang; Costa, Maria; Bassi, Gurminder; Lai, Yiu-Kay; Michel, François

    2011-01-01

    A survey of sequence databases revealed 10 instances of subgroup IIB1 mitochondrial ribosomal introns with 1 to 33 additional nucleotides inserted between the 5′ exon and the consensus sequence at the intron 5′ end. These 10 introns depart further from the IIB1 consensus in their predicted domain VI structure: In contrast to its basal helix and distal GNRA terminal loop, the middle part of domain VI is highly variable and lacks the bulging A that serves as the branchpoint in lariat formation....

  19. Application of DNA Bar Codes for Screening of Industrially Important Fungi: the Haplotype of Trichoderma harzianum Sensu Stricto Indicates Superior Chitinase Formation▿

    OpenAIRE

    Nagy, Viviana; Seidl, Verena; Szakacs, George; Komoń-Zelazowska, Monika; Kubicek, Christian P; Irina S. Druzhinina

    2007-01-01

    Selection of suitable strains for biotechnological purposes is frequently a random process supported by high-throughput methods. Using chitinase production by Hypocrea lixii/Trichoderma harzianum as a model, we tested whether fungal strains with superior enzyme formation may be diagnosed by DNA bar codes. We analyzed sequences of two phylogenetic marker loci, internal transcribed spacer 1 (ITS1) and ITS2 of the rRNA-encoding gene cluster and the large intron of the elongation factor 1-alpha g...

  20. Extraction of Crude Chitinase from Higher Plants and their Chitin-Hydrolysis Activities; Kotosyokubutu yurai kichinaze no chusyutu to kichin bunkai kassei

    Energy Technology Data Exchange (ETDEWEB)

    Kondo, K.; Harada, K.; Shibata, M.; Maeda, R. [Doshisha Univ., Kyoto (Japan). Faculty of Engineering

    1997-07-10

    To prepare a purified chitinase from higher plants, firstly, crude enzymes were extracted from six higher plants, namely, radish seeds, sunflower seeds, watermelon seeds, bamboo leaves, orange skin, and persimmon skin. Using these crude enzymes, pH dependencies of hydrolysis reaction of colloidal chitin are investigated. For radish seeds and bamboo leaves, which have relatively high activities, the kinetics of enzymatic reaction are studies. It is clear that these reactions obey Michaelis-Menten kinetics. 7 refs., 3 figs., 2 tabs.

  1. Urinary chitinase 3-like protein 1 for early diagnosis of acute kidney injury: a prospective cohort study in adult critically ill patients

    OpenAIRE

    De Loor, Jorien; Decruyenaere, Johan; Demeyere, Kristel; Nuytinck, Lieve; Hoste, Eric AJ; Meyer, Evelyne

    2016-01-01

    Background Acute kidney injury (AKI) occurs frequently and adversely affects patient and kidney outcomes, especially when its severity increases from stage 1 to stages 2 or 3. Early interventions may counteract such deterioration, but this requires early detection. Our aim was to evaluate whether the novel renal damage biomarker urinary chitinase 3-like protein 1 (UCHI3L1) can detect AKI stage ≥2 more early than serum creatinine and urine output, using the respective Kidney Disease | Improvin...

  2. Effect of Botrytis cinerea infection and elicitation on ß-1,3-glucanase and chitinase activity in bean leaves and cell cultures

    OpenAIRE

    Elżbieta Kuźniak; Henryk Urbanek; Aneta Michalak; Katarzyna Herka

    2013-01-01

    The activity of ß-1,3-glucanase and chitinase in bean plants treated with B. cinerea products or/and infected and in cell cultures after application of fungal products has been studied. Botrytis cinerea infection and culture filtrates, ethanol precipitates, glucan and conidial extract treatment markedly enhanced the activity of both hydrolases. Cell cultures treated with B.cinerea products reacted similarly to intact plants. In plants pretreated with 2-day culture filtrate and conidial extrac...

  3. Turnabout Is Fair Play: Herbivory-Induced Plant Chitinases Excreted in Fall Armyworm Frass Suppress Herbivore Defenses in Maize1[OPEN

    Science.gov (United States)

    Alves, Patrick C.M.S.; Gaffoor, Iffa; Acevedo, Flor E.; Peiffer, Michelle; Jin, Shan; Han, Yang; Shakeel, Samina; Felton, Gary W.

    2016-01-01

    The perception of herbivory by plants is known to be triggered by the deposition of insect-derived factors such as saliva and oral secretions, oviposition materials, and even feces. Such insect-derived materials harbor chemical cues that may elicit herbivore and/or pathogen-induced defenses in plants. Several insect-derived molecules that trigger herbivore-induced defenses in plants are known; however, insect-derived molecules suppressing them are largely unknown. In this study, we identified two plant chitinases from fall armyworm (Spodoptera frugiperda) larval frass that suppress herbivore defenses while simultaneously inducing pathogen defenses in maize (Zea mays). Fall armyworm larvae feed in enclosed whorls of maize plants, where frass accumulates over extended periods of time in close proximity to damaged leaf tissue. Our study shows that maize chitinases, Pr4 and Endochitinase A, are induced during herbivory and subsequently deposited on the host with the feces. These plant chitinases mediate the suppression of herbivore-induced defenses, thereby increasing the performance of the insect on the host. Pr4 and Endochitinase A also trigger the antagonistic pathogen defense pathway in maize and suppress fungal pathogen growth on maize leaves. Frass-induced suppression of herbivore defenses by deposition of the plant-derived chitinases Pr4 and Endochitinase A is a unique way an insect can co-opt the plant’s defense proteins for its own benefit. It is also a phenomenon unlike the induction of herbivore defenses by insect oral secretions in most host-herbivore systems. PMID:26979328

  4. Recurrent insertion of 5'-terminal nucleotides and loss of the branchpoint motif in lineages of group II introns inserted in mitochondrial preribosomal RNAs.

    Science.gov (United States)

    Li, Cheng-Fang; Costa, Maria; Bassi, Gurminder; Lai, Yiu-Kay; Michel, François

    2011-07-01

    A survey of sequence databases revealed 10 instances of subgroup IIB1 mitochondrial ribosomal introns with 1 to 33 additional nucleotides inserted between the 5' exon and the consensus sequence at the intron 5' end. These 10 introns depart further from the IIB1 consensus in their predicted domain VI structure: In contrast to its basal helix and distal GNRA terminal loop, the middle part of domain VI is highly variable and lacks the bulging A that serves as the branchpoint in lariat formation. In vitro experiments using two closely related IIB1 members inserted at the same ribosomal RNA site in the basidiomycete fungi Grifola frondosa and Pycnoporellus fulgens revealed that both ribozymes are capable of efficient self-splicing. However, whereas the Grifola intron was excised predominantly as a lariat, the Pycnoporellus intron, which possesses six additional nucleotides at the 5' end, yielded only linear products, consistent with its predicted domain VI structure. Strikingly, all of the introns with 5' terminal insertions lack the EBS2 exon-binding site. Moreover, several of them are part of the small subset of group II introns that encode potentially functional homing endonucleases of the LAGLIDADG family rather than reverse transcriptases. Such coincidences suggest causal relationships between the shift to DNA-based mobility, the loss of one of the two ribozyme sites for binding the 5' exon, and the exclusive use of hydrolysis to initiate splicing. PMID:21613530

  5. Functional Characterization of Novel Chitinase Genes Present in the Sheath Blight Resistance QTL: qSBR11-1 in Rice Line Tetep.

    Science.gov (United States)

    Richa, Kamboj; Tiwari, Ila M; Kumari, Mandeep; Devanna, B N; Sonah, Humira; Kumari, Archana; Nagar, Ramawatar; Sharma, Vinay; Botella, Jose R; Sharma, Tilak R

    2016-01-01

    Rice sheath blight disease caused by Rhizoctonia solani is one of the most devastating diseases in rice leading to heavy yield losses. Due to the polygenic nature of resistance, no major resistance gene with complete host resistance against R. solani has been reported. In this study, we have performed molecular and functional analysis of the genes associated with the major R. solani-resistance QTL qSBR11-1 in the indica rice line Tetep. Sequence analysis revealed the presence of a set of 11 tandem repeats containing genes with a high degree of homology to class III chitinase defense response genes. Real-time quantitative PCR analysis showed that all the genes are strongly induced 36 h after R. solani infection. Comparison between the resistant Tetep and the susceptible HP2216 lines shows that the induction of the chitinase genes is much higher in the Tetep line. Recombinant protein produced in vitro for six of the eleven genes showed chitinolytic activity in gel assays but we did not detect any xylanase inhibitory activity. All the six in vitro expressed proteins show antifungal activity with a clear inhibitory effect on the growth of the R. solani mycelium. The characterized chitinase genes can provide an important resource for the genetic improvement of R. solani susceptible rice lines for sheath blight resistance breeding. PMID:26973685

  6. Isolation and Molecular Characterization of Chitinase-Deficient Bacillus licheniformis Strains Capable of Deproteinization of Shrimp Shell Waste To Obtain Highly Viscous Chitin▿

    Science.gov (United States)

    Waldeck, Jens; Daum, Gabriele; Bisping, Bernward; Meinhardt, Friedhelm

    2006-01-01

    Proteolytic but chitinase-deficient microbial cultures were isolated from shrimp shell waste and characterized. The most efficient isolate was found to be a mixed culture consisting of two Bacillus licheniformis strains, which were first determined microscopically and physiologically. Molecular characterization was carried out by sequencing the 16S rRNA gene of both strains. According to the residual protein and ash content, the chitin obtained by fermentation of such a mixed culture was found to be comparable to a commercially available, chemically processed product. However, the strikingly high viscosity (80 versus 10 mPa of the commercially available sample) indicates its superior quality. The two strains differed in colony morphology and in their secretion capabilities for degradative extracellular enzymes. Sequencing of the loci encoding amylase, cellulase, chitinases, and proteases, as well as the degS/degU operon, which is instrumental in the regulation of degradative enzymes, and the pga operon, which is responsible for polyglutamic acid production, revealed no differences. However, a frameshift mutation in chiA, encoding a chitinase, was validated for both strains, providing an explanation for the ascertained absence of chitinolytic activities and the concomitant possibility of producing highly viscous chitin in a fermentational deproteinization process. PMID:17028230

  7. AC loss performance of CS insert coil

    International Nuclear Information System (INIS)

    The ITER Central Solenoid (CS) model coil and the CS insert coil were fabricated, and the test was carried out. The AC loss measurement of the coil is one of the most important tests to determine coil performance. The AC loss of a short sample conductor for the CS insert coil was measured by using the calorimetric method, and the coupling time constants of the conductor were estimated to be 30 ms and 20 ms for pulse and discharge tests, respectively. The AC loss of the CS insert coil was measured by using the calorimetric method for pulse and discharge tests. The coupling time constant estimated from the result of the pulse tests was 34 ms and almost equal to that of the short sample. The coupling time constant for the discharge test was estimated to be 140 ms and about 4 times that of the pulse test. (author)

  8. Involvement of the MAPK and PI3K pathways in chitinase 3-like 1-regulated hyperoxia-induced airway epithelial cell death

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Mi Na; Lee, Kyung Eun; Hong, Jung Yeon; Heo, Won Il; Kim, Kyung Won; Kim, Kyu Earn [Department of Pediatrics and Institute of Allergy, Severance Medical Research Institute, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul (Korea, Republic of); Sohn, Myung Hyun, E-mail: mhsohn@yuhs.ac [Department of Pediatrics and Institute of Allergy, Severance Medical Research Institute, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul (Korea, Republic of)

    2012-05-18

    Highlights: Black-Right-Pointing-Pointer Hyperoxia induces apoptosis and chitinase 3-like 1 expression in human airway epithelial cells. Black-Right-Pointing-Pointer Presence of chitinase 3-like 1 affects airway epithelial cell death after hyperoxic exposure. Black-Right-Pointing-Pointer Silencing chitinase 3-like 1 manipulate the phosphorylation of ERK, p38 and Akt. -- Abstract: Background: Exposure to 100% oxygen causes hyperoxic acute lung injury characterized by cell death and injury of alveolar epithelial cells. Recently, the role of chitinase 3-like 1 (CHI3L1), a member of the glycosyl hydrolase 18 family that lacks chitinase activity, in oxidative stress was demonstrated in murine models. High levels of serum CHI3L1 have been associated with various diseases of the lung, such as asthma, chronic obstructive pulmonary disease, and cancer. However, the role of CHI3L1 in human airway epithelial cells undergoing oxidative stress remains unknown. In addition, the signaling pathways associated with CHI3L1 in this process are poorly understood. Purpose: In this study, we demonstrate the role of CHI3L1, along with the MAPK and PI3K signaling pathways, in hyperoxia-exposed airway epithelial cells. Method: The human airway epithelial cell line, BEAS-2B, was exposed to >95% oxygen (hyperoxia) for up to 72 h. Hyperoxia-induced cell death was determined by assessing cell viability, Annexin-V FITC staining, caspase-3 and -7 expression, and electron microscopy. CHI3L1 knockdown and overexpression studies were conducted in BEAS-2B cells to examine the role of CHI3L1 in hyperoxia-induced apoptosis. Activation of the MAPK and PI3K pathways was also investigated to determine the role of these signaling cascades in this process. Results: Hyperoxia exposure increased CHI3L1 expression and apoptosis in a time-dependent manner. CHI3L1 knockdown protected cells from hyperoxia-induced apoptosis. In contrast, CHI3L1 overexpression promoted cell death after hyperoxia exposure. Finally

  9. RERTR-12 Insertion 2 Irradiation Summary Report

    Energy Technology Data Exchange (ETDEWEB)

    D. M. Perez; G. S. Chang; D. M. Wachs; G. A. Roth; N. E. Woolstenhulme

    2012-09-01

    The Reduced Enrichment for Research and Test Reactor (RERTR) experiment RERTR-12 was designed to provide comprehensive information on the performance of uranium-molybdenum (U-Mo) based monolithic fuels for research reactor applications.1 RERTR-12 insertion 2 includes the capsules irradiated during the last three irradiation cycles. These capsules include Z, Y1, Y2 and Y3 type capsules. The following report summarizes the life of the RERTR-12 insertion 2 experiment through end of irradiation, including as-run neutronic analysis results, thermal analysis results and hydraulic testing results.

  10. Active interlock for storage ring insertion devices

    International Nuclear Information System (INIS)

    The insertion devices in the NSLS x-ray ring produce such intense photon beams that passive measures alone are not adequate to protect the vacuum chamber. An active interlock is used to prevent thermal damage from improperly steered electron beams. The interlock system measures the position of the electron beam on both sides of the insertion devices using pick-up electrodes and interrupts the rf if the beam moves outside a safe window. The interlock features redundant circuitry as well as an automatic testing procedure. 1 ref., 4 figs

  11. Installation of the LHC experimental insertions

    CERN Document Server

    Bartolome-Jimenez, S

    2004-01-01

    The installation of the LHC experimental insertions, and particularly the installation of the low-beta quadrupoles, raises many technical challenges due to the stringent alignment specifications and to the difficulty of access in very confined areas. The compact layout with many lattice elements, vacuum components, beam control instrumentation and the presence of shielding does not allow for any improvisation in the installation procedure. This paper reviews all the constraints that need to be taken into account when installing the experimental insertions. It describes the chronological sequence of installation and discusses the technical solutions that have been adopted.

  12. INSTALLATION OF THE LHC EXPERIMENTAL INSERTIONS

    CERN Document Server

    Bartolome-Jimenez, S

    2004-01-01

    The installation of the LHC experimental insertions, and particularly the installation of the Low-Beta quadrupoles, raises many technical challenges due to the stringent alignment specifications and to the difficulty of access in very confined areas. The compact layout with many lattice elements, vacuum components, beam control instrumentation and the presence of shielding does not allow for any improvisation in the installation procedure. This paper reviews all the constraints that need to be taken into account when installing the experimental insertions. It describes the chronological sequence of installation and discusses the technical solutions that have been adopted.

  13. Allosteric Switching by Mutually Exclusive Folding of Protein Domains

    OpenAIRE

    Radley, Tracy L.; Markowska, Anna I.; Bettinger, Blaine T.; Ha, Jeung-Hoi; Loh, Stewart N.

    2003-01-01

    Many proteins are built from structurally and functionally distinct and domains. A major goal is to understand how conformational change transmits information between domains in order to achieve biological activity. A two-domain, bi-functional fusion protein has been designed so that the mechanical stress imposed by the folded structure of one subunit causes the other subunit to unfold, and vice versa. The construct consists of ubiquitin inserted into a surface loop of barnase. The distance b...

  14. Trusted Domain

    DEFF Research Database (Denmark)

    Hjorth, Theis Solberg; Torbensen, Rune

    2012-01-01

    remote access via IP-based devices such as smartphones. The Trusted Domain platform fits existing legacy technologies by managing their interoperability and access controls, and it seeks to avoid the security issues of relying on third-party servers outside the home. It is a distributed system that......In the digital age of home automation and with the proliferation of mobile Internet access, the intelligent home and its devices should be accessible at any time from anywhere. There are many challenges such as security, privacy, ease of configuration, incompatible legacy devices, a wealth of...

  15. Structural and Histone Binding Ability Characterizations of Human PWWP Domains

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Hong; Zeng, Hong; Lam, Robert; Tempel, Wolfram; Amaya, Maria F.; Xu, Chao; Dombrovski, Ludmila; Qiu, Wei; Wang, Yanming; Min, Jinrong (Toronto); (Penn)

    2013-09-25

    The PWWP domain was first identified as a structural motif of 100-130 amino acids in the WHSC1 protein and predicted to be a protein-protein interaction domain. It belongs to the Tudor domain 'Royal Family', which consists of Tudor, chromodomain, MBT and PWWP domains. While Tudor, chromodomain and MBT domains have long been known to bind methylated histones, PWWP was shown to exhibit histone binding ability only until recently. The PWWP domain has been shown to be a DNA binding domain, but sequence analysis and previous structural studies show that the PWWP domain exhibits significant similarity to other 'Royal Family' members, implying that the PWWP domain has the potential to bind histones. In order to further explore the function of the PWWP domain, we used the protein family approach to determine the crystal structures of the PWWP domains from seven different human proteins. Our fluorescence polarization binding studies show that PWWP domains have weak histone binding ability, which is also confirmed by our NMR titration experiments. Furthermore, we determined the crystal structures of the BRPF1 PWWP domain in complex with H3K36me3, and HDGF2 PWWP domain in complex with H3K79me3 and H4K20me3. PWWP proteins constitute a new family of methyl lysine histone binders. The PWWP domain consists of three motifs: a canonical {beta}-barrel core, an insertion motif between the second and third {beta}-strands and a C-terminal {alpha}-helix bundle. Both the canonical {beta}-barrel core and the insertion motif are directly involved in histone binding. The PWWP domain has been previously shown to be a DNA binding domain. Therefore, the PWWP domain exhibits dual functions: binding both DNA and methyllysine histones.

  16. Chicken models of retroviral insertional mutagenesis

    Czech Academy of Sciences Publication Activity Database

    Pečenka, Vladimír; Karafiát, Vít; Dvořák, Michal

    New York: Springer, 2011 - (Dupuy, A.; Largaespada, D.), s. 77-112 ISBN 978-1-4419-7655-0 R&D Projects: GA ČR GA301/09/1727 Institutional research plan: CEZ:AV0Z50520514 Keywords : insertional mutagenesis * chicken model * MAV retroviruses Subject RIV: EB - Genetics ; Molecular Biology

  17. Thermal Performance of the XRS Helium Insert

    Science.gov (United States)

    Breon, Susan R.; DiPirro, Michael J.; Tuttle, James G.; Shirron, Peter J.; Warner, Brent A.; Boyle, Robert F.; Canavan, Edgar R.

    1999-01-01

    The X-Ray Spectrometer (XRS) is an instrument on the Japanese Astro-E satellite, scheduled for launch early in the year 2000. The XRS Helium Insert comprises a superfluid helium cryostat, an Adiabatic Demagnetization Refrigerator (ADR), and the XRS calorimeters with their cold electronics. The calorimeters are capable of detecting X-rays over the energy range 0.1 to 10 keV with a resolution of 12 eV. The Helium Insert completed its performance and verification testing at Goddard in January 1999. It was shipped to Japan, where it has been integrated with the neon dewar built by Sumitomo Heavy Industries. The Helium Insert was given a challenging lifetime requirement of 2.0 years with a goal of 2.5 years. Based on the results of the thermal performance tests, the predicted on-orbit lifetime is 2.6 years with a margin of 30%. This is the result of both higher efficiency in the ADR cycle and the low temperature top-off, more than compensating for an increase in the parasitic heat load. This paper presents a summary of the key design features and the results of the thermal testing of the XRS Helium Insert.

  18. Chicken models of retroviral insertional mutagenesis

    Czech Academy of Sciences Publication Activity Database

    Pečenka, Vladimír; Karafiát, Vít; Dvořák, Michal

    New York : Springer, 2011 - (Dupuy, A.; Largaespada, D.), s. 77-112 ISBN 978-1-4419-7655-0 R&D Projects: GA ČR GA301/09/1727 Institutional research plan: CEZ:AV0Z50520514 Keywords : insertional mutagenesis * chicken model * MAV retroviruses Subject RIV: EB - Genetics ; Molecular Biology

  19. A design for vertical crossing insertions

    Energy Technology Data Exchange (ETDEWEB)

    Garren, A.

    1985-10-01

    A crossing insertion designed for an SSC with vertically separated 1-in-1 beam lines is presented in this note. The author supposes that the beam lines consist of separate magnets in separate cryostats separated by about 70 cm. He then describes the design, where vertical separation is done with four vertical dipoles producing a steplike beam line.

  20. Functional characterization of Mammary Gland Protein-40, a chitinase-like glycoprotein expressed during mammary gland apoptosis.

    Science.gov (United States)

    Anand, Vijay; Jaswal, Shalini; Singh, Surender; Kumar, Sudarshan; Jena, Manoj Kumar; Verma, Arvind Kumar; Yadav, Munna Lal; Janjanam, Jagadeesh; Lotfan, Masoud; Malakar, Dhruba; Dang, Ajay Kumar; Mohanty, Tushar Kumar; Kaushik, Jai Kumar; Mohanty, Ashok Kumar

    2016-02-01

    MGP-40 is a chitinase-like protein which is over expressed during mammary gland involution. However, its physiological function in the mammary gland is poorly understood. In the present investigation, we have reported the functional significance of buffalo specific MGP-40 in the mammary gland by using an in vitro model of the buffalo mammary epithelial cell (BuMEC) line. MGP-40 was highly up regulated in BuMECs in serum starved condition as well as after treatment with prolactin suggesting its role in the stress response. Subsequently, to study the effect of MGP-40 on BuMECs, the cells were transfected with a mammalian expression construct of pCI neo harboring MGP-40 gene. It was observed that over expression of MGP-40 enhanced proliferation of BuMECs and protected the cells from apoptosis under serum free condition. In contrast, MGP-40 attenuated the mitogenic effect of insulin in BuMECs. Besides, over expression of the MGP-40 reduced dome formation, acinar polarization and casein synthesis in BuMECs in the presence of lactogenic hormones, it also induced Stat3 phosphorylation and epithelial to mesenchymal transition (EMT) -like features. Together, our data suggest that MGP-40 is involved in protection of BuMECs under stress conditions, inhibits cellular differentiation and induces EMT-like features. A schematic diagram depicting possible association of MGP-40 in various molecular pathways has been presented. PMID:26659075

  1. A small RNA controls expression of the chitinase ChiA in Listeria monocytogenes

    DEFF Research Database (Denmark)

    Nielsen, Jesper S; Larsen, Marianne Halberg; Lillebæk, Eva Maria Sternkopf;

    2011-01-01

    In recent years, more than 60 small RNAs (sRNAs) have been identified in the gram-positive human pathogen Listeria monocytogenes, but their putative roles and mechanisms of action remain largely unknown. The sRNA LhrA was recently shown to be a post-transcriptional regulator of a single gene, lmo...... role of LhrA in L. monocytogenes. To this end, we determined the effects of LhrA on global-wide gene expression. We observed that nearly 300 genes in L. monocytogenes are either positively or negatively affected by LhrA. Among these genes, we identified lmo0302 and chiA as direct targets of LhrA, thus...... establishing LhrA as a multiple target regulator. Lmo0302 encodes a hypothetical protein with no known function, whereas chiA encodes one of two chitinases present in L. monocytogenes. We show here that LhrA acts as a post-transcriptional regulator of lmo0302 and chiA by interfering with ribosome recruitment...

  2. Biochemical characterization of a recombinant plant class III chitinase from the pitcher of the carnivorous plant Nepenthes alata.

    Science.gov (United States)

    Ishisaki, Kana; Arai, Sachiko; Hamada, Tatsuro; Honda, Yuji

    2012-11-01

    A class III chitinase belonging to the GH18 family from Nepenthes alata (NaCHIT3) was expressed in Escherichia coli. The enzyme exhibited hydrolytic activity toward colloidal chitin, ethylene glycol chitin, and (GlcNAc)(n) (n=5 and 6). The enzyme hydrolyzed the fourth glycosidic linkage from the non-reducing end of (GlcNAc)(6). The anomeric form of the products indicated it was a retaining enzyme. The colloidal chitin hydrolytic reaction displayed high activity between pH 3.9 and 6.9, but the pH optimum of the (GlcNAc)(6) hydrolytic reaction was 3.9 at 37 °C. The optimal temperature for activity was 65 °C in 50 mM sodium acetate buffer (pH 3.9). The pH optima of NaCHIT3 and NaCHIT1 might be related to their roles in chitin degradation in the pitcher fluid. PMID:23026711

  3. Effect of mouse antisera targeting the Phlebotomus papatasi midgut chitinase PpChit1 on sandfly physiology and fitness

    Directory of Open Access Journals (Sweden)

    Maricela Robles-Murguia

    2014-12-01

    Full Text Available In sandflies, the absence of the peritrophic matrix (PM affects the rate of blood digestion. Also, the kinetics of PM secretion varies according to species. We previously characterised PpChit1, a midgut-specific chitinase secreted in Phlebotomus papatasi (PPIS that is involved in the maturation of the PM and showed that antibodies against PpChit1 reduce the chitinolytic activity in the midgut of several sandfly species. Here, sandflies were fed on red blood cells reconstituted with naïve or anti-PpChit1 sera and assessed for fitness parameters that included blood digestion, oviposition onset, number of eggs laid, egg bouts, average number of eggs per bout and survival. In PPIS, anti-PpChit1 led to a one-day delay in the onset of egg laying, with flies surviving three days longer compared to the control group. Anti-PpChit1 also had a negative effect on overall ability of flies to lay eggs, as several gravid females from all three species were unable to lay any eggs despite having lived longer than control flies. Whereas the longer survival might be associated with improved haeme scavenging ability by the PM, the inability of females to lay eggs is possibly linked to changes in PM permeability affecting nutrient absorption.

  4. Mining of unexplored habitats for novel chitinases--chiA as a helper gene proxy in metagenomics.

    Science.gov (United States)

    Cretoiu, Mariana Silvia; Kielak, Anna Maria; Abu Al-Soud, Waleed; Sørensen, Søren J; van Elsas, Jan Dirk

    2012-06-01

    The main objective of this study was to assess the abundance and diversity of chitin-degrading microbial communities in ten terrestrial and aquatic habitats in order to provide guidance to the subsequent exploration of such environments for novel chitinolytic enzymes. A combined protocol which encompassed (1) classical overall enzymatic assays, (2) chiA gene abundance measurement by qPCR, (3) chiA gene pyrosequencing, and (4) chiA gene-based PCR-DGGE was used. The chiA gene pyrosequencing is unprecedented, as it is the first massive parallel sequencing of this gene. The data obtained showed the existence across habitats of core bacterial communities responsible for chitin assimilation irrespective of ecosystem origin. Conversely, there were habitat-specific differences. In addition, a suite of sequences were obtained that are as yet unregistered in the chitinase database. In terms of chiA gene abundance and diversity, typical low-abundance/diversity versus high-abundance/diversity habitats was distinguished. From the combined data, we selected chitin-amended agricultural soil, the rhizosphere of the Arctic plant Oxyria digyna and the freshwater sponge Ephydatia fluviatilis as the most promising habitats for subsequent bioexploration. Thus, the screening strategy used is proposed as a guide for further metagenomics-based exploration of the selected habitats. PMID:22526805

  5. Screw-Thread Inserts As Temporary Flow Restrictors

    Science.gov (United States)

    Trimarchi, Paul

    1992-01-01

    Coil-spring screw-thread inserts found useful as temporary flow restrictors. Inserts placed in holes through which flow restricted, effectively reducing cross sections available for flow. Friction alone holds inserts against moderate upstream pressures. Use of coil-spring thread inserts as flow restrictors conceived as inexpensive solution to problem of adjusting flow of oxygen through orifices in faceplate into hydrogen/oxygen combustion chamber. Installation and removal of threaded inserts gentle enough not to deform orifice tubes.

  6. Comparison of conventional Injection Mould Inserts to Additively Manufactured Inserts using Life Cycle Assessment

    DEFF Research Database (Denmark)

    Hofstätter, Thomas; Bey, Niki; Mischkot, Michael;

    2016-01-01

    Polymer Additive Manufacturing can be used to produce soft tooling inserts for injection moulding. Compared to conventional tooling, the energy and time consumption during production are significantly lower. As the life time of such inserts is significantly shorter than the life time of traditional...... potential environmental impact and yield throughout the development and pilot phase. Insert geometry is particularly advantageous for pilot production and small production sizes. In this research, Life Cycle Assessment is used to compare the environmental impact of soft tooling by Additive Manufacturing...

  7. Interaction of nuclease colicins with membranes: insertion depth correlates with bilayer perturbation.

    Directory of Open Access Journals (Sweden)

    Mireille Vankemmelbeke

    Full Text Available BACKGROUND: Protein transport across cellular membranes is an important aspect of toxin biology. Escherichia coli cell killing by nuclease colicins occurs through DNA (DNases or RNA (RNases hydrolysis and to this end their cytotoxic domains require transportation across two sets of membranes. In order to begin to unravel the molecular mechanisms underlying the membrane translocation of colicin nuclease domains, we have analysed the membrane association of four DNase domains (E9, a charge reduction E9 mutant, E8, and E7 and one ribosomal RNase domain (E3 using a biomembrane model system. PRINCIPAL RESULTS: We demonstrate, through the use of large unilamellar vesicles composed of synthetic and E. coli lipids and a membrane surface potential sensor, that the colicin nuclease domains bind anionic membranes only, with micromolar affinity and via a cooperative binding mechanism. The evaluation of the nuclease bilayer insertion depth, through a fluorescence quenching analysis using brominated lipids, indicates that the nucleases locate to differential regions in the bilayer. Colicin DNases target the interfacial region of the lipid bilayer, with the DNase E7 showing the deepest insertion, whereas the ribosomal RNase E3 penetrates into the hydrophobic core region of the bilayer. Furthermore, the membrane association of the DNase E7 and the ribosomal RNase E3 induces vesicle aggregation, lipid mixing and content leakage to a much larger extent than that of the other DNases analysed. CONCLUSIONS/SIGNIFICANCE: Our results show, for the first time, that after the initial electrostatically driven membrane association, the pleiotropic membrane effects induced by colicin nuclease domains relate to their bilayer insertion depth and may be linked to their in vivo membrane translocation.

  8. Membrane topology and insertion of membrane proteins: Search for topogenic signals

    OpenAIRE

    van Geest, Marleen; Lolkema, Juke S.

    2000-01-01

    Integral membrane proteins are found in all cellular membranes and carry out many of the functions that are essential to life. The membrane-embedded domains of integral membrane proteins are structurally quite simple, allowing the use of various prediction methods and biochemical methods to obtain structural information about membrane proteins. A critical step in the biosynthetic pathway leading to the folded protein in the membrane is ifs insertion into the lipid bilayer. Understanding of th...

  9. MORPH INSERTION AND ALLOMORPHY IN OPTIMALITY THEORY

    OpenAIRE

    Eulalia Bonet

    2004-01-01

    The goal of this paper is to compare two different hypotheses about the insertion of morphs and allomorphy in Optimality Theory. One of them, the Morphs through Constraifits Hypothesis (MCH) claims that the phonological realization of morphemes (morphs) is introduced through language-particular constraints. The other hypothesis, the Morphs in the Input Hypothesis (MIH) claims that the inputs to GEN contain al1 the relevant phonological information. It is shown that the MIH is clearly superior...

  10. Beamline Insertions Manager at Jefferson Lab

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Michael C. [Jefferson Lab., Newport News, VA (United States)

    2015-09-01

    The beam viewer system at Jefferson Lab provides operators and beam physicists with qualitative and quantitative information on the transverse electron beam properties. There are over 140 beam viewers installed on the 12 GeV CEBAF accelerator. This paper describes an upgrade consisting of replacing the EPICS-based system tasked with managing all viewers with a mixed system utilizing EPICS and high-level software. Most devices, particularly the beam viewers, cannot be safely inserted into the beam line during high-current beam operations. Software is partly responsible for protecting the machine from untimely insertions. The multiplicity of beam-blocking and beam-vulnerable devices motivates us to try a data-driven approach. The beamline insertions application components are centrally managed and configured through an object-oriented software framework created for this purpose. A rules-based engine tracks the configuration and status of every device, along with the beam status of the machine segment containing the device. The application uses this information to decide on which device actions are allowed at any given time.

  11. Roles for retrotransposon insertions in human disease.

    Science.gov (United States)

    Hancks, Dustin C; Kazazian, Haig H

    2016-01-01

    Over evolutionary time, the dynamic nature of a genome is driven, in part, by the activity of transposable elements (TE) such as retrotransposons. On a shorter time scale it has been established that new TE insertions can result in single-gene disease in an individual. In humans, the non-LTR retrotransposon Long INterspersed Element-1 (LINE-1 or L1) is the only active autonomous TE. In addition to mobilizing its own RNA to new genomic locations via a "copy-and-paste" mechanism, LINE-1 is able to retrotranspose other RNAs including Alu, SVA, and occasionally cellular RNAs. To date in humans, 124 LINE-1-mediated insertions which result in genetic diseases have been reported. Disease causing LINE-1 insertions have provided a wealth of insight and the foundation for valuable tools to study these genomic parasites. In this review, we provide an overview of LINE-1 biology followed by highlights from new reports of LINE-1-mediated genetic disease in humans. PMID:27158268

  12. Fungus- and wound-induced accumulation of mRNA containing a class II chitinase of the pathogenesis-related protein 4 (PR-4) family of maize.

    Science.gov (United States)

    Bravo, Juan Manuel; Campo, Sonia; Murillo, Isabel; Coca, Mária; San Segundo, Blanca

    2003-07-01

    Pathogenesis-related (PR) proteins are plant proteins that are induced in response to pathogen attack. PR proteins are grouped into independent families based on their sequences and properties. The PR-4 family comprises class I and class II chitinases. We have isolated a full-length cDNA encoding a chitinase from maize which shares a high degree of nucleotide and amino acid sequence homology with the class II chitinases of the PR-4 family of PR proteins. Our results indicate that fungal infection, and treatment either with fungal elicitors or with moniliformin, a mycotoxin produced by the fungus Fusarium moniliforme, increase the level of ZmPR4 mRNA. In situ mRNA hybridization analysis in sections obtained from fungus-infected germinating embryos revealed that ZmPR4 mRNA accumulation occurs in those cell types that first establish contact with the pathogen. ZmPR4 mRNA accumulation is also stimulated by treatment with silver nitrate whereas the application of the hormones gibberellic acid or acetylsalicylic acid has no effect. Wounding, or treatment with abscisic acid or methyl jasmonate, results in accumulation of ZmPR4 mRNA in maize leaves. Furthermore, the ZmPR4 protein was expressed in Escherichia coli, purified and used to obtain polyclonal antibodies that specifically recognized ZmPR4 in protein extracts from fungus-infected embryos. Accumulation of ZmPR4 mRNA in fungus-infected maize tissues was accompanied by a significant accumulation of the corresponding protein. The possible implications of these findings as part of the general defence response of maize plants against pathogens are discussed. PMID:13677464

  13. Glucanases and chitinases as causal agents in the protection of Acacia extrafloral nectar from infestation by phytopathogens.

    Science.gov (United States)

    González-Teuber, Marcia; Pozo, María J; Muck, Alexander; Svatos, Ales; Adame-Alvarez, Rosa M; Heil, Martin

    2010-03-01

    Nectars are rich in primary metabolites and attract mutualistic animals, which serve as pollinators or as an indirect defense against herbivores. Their chemical composition makes nectars prone to microbial infestation. As protective strategy, floral nectar of ornamental tobacco (Nicotiana langsdorffii x Nicotiana sanderae) contains "nectarins," proteins producing reactive oxygen species such as hydrogen peroxide. By contrast, pathogenesis-related (PR) proteins were detected in Acacia extrafloral nectar (EFN), which is secreted in the context of defensive ant-plant mutualisms. We investigated whether these PR proteins protect EFN from phytopathogens. Five sympatric species (Acacia cornigera, A. hindsii, A. collinsii, A. farnesiana, and Prosopis juliflora) were compared that differ in their ant-plant mutualism. EFN of myrmecophytes, which are obligate ant-plants that secrete EFN constitutively to nourish specialized ant inhabitants, significantly inhibited the growth of four out of six tested phytopathogenic microorganisms. By contrast, EFN of nonmyrmecophytes, which is secreted only transiently in response to herbivory, did not exhibit a detectable inhibitory activity. Combining two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with nanoflow liquid chromatography-tandem mass spectrometry analysis confirmed that PR proteins represented over 90% of all proteins in myrmecophyte EFN. The inhibition of microbial growth was exerted by the protein fraction, but not the small metabolites of this EFN, and disappeared when nectar was heated. In-gel assays demonstrated the activity of acidic and basic chitinases in all EFNs, whereas glucanases were detected only in EFN of myrmecophytes. Our results demonstrate that PR proteins causally underlie the protection of Acacia EFN from microorganisms and that acidic and basic glucanases likely represent the most important prerequisite in this defensive function. PMID:20023149

  14. Expression of the chitinase family glycoprotein YKL-40 in undifferentiated, differentiated and trans-differentiated mesenchymal stem cells.

    Directory of Open Access Journals (Sweden)

    Daniel J Hoover

    Full Text Available The glycoprotein YKL-40 (CHI3L1 is a secreted chitinase family protein that induces angiogenesis, cell survival, and cell proliferation, and plays roles in tissue remodeling and immune regulation. It is expressed primarily in cells of mesenchymal origin, is overexpressed in numerous aggressive carcinomas and sarcomas, but is rarely expressed in normal ectodermal tissues. Bone marrow-derived mesenchymal stem cells (MSCs can be induced to differentiate into various mesenchymal tissues and trans-differentiate into some non-mesenchymal cell types. Since YKL-40 has been used as a mesenchymal marker, we followed YKL-40 expression as undifferentiated MSCs were induced to differentiate into bone, cartilage, and neural phenotypes. Undifferentiated MSCs contain significant levels of YKL-40 mRNA but do not synthesize detectable levels of YKL-40 protein. MSCs induced to differentiate into chondrocytes and osteocytes soon began to express and secrete YKL-40 protein, as do ex vivo cultured chondrocytes and primary osteocytes. In contrast, MSCs induced to trans-differentiate into neurons did not synthesize YKL-40 protein, consistent with the general absence of YKL-40 protein in normal CNS parenchyma. However, these trans-differentiated neurons retained significant levels of YKL-40 mRNA, suggesting the mechanisms which prevented YKL-40 translation in undifferentiated MSCs remained in place, and that these trans-differentiated neurons differ in at least this way from neurons derived from neuronal stem cells. Utilization of a differentiation protocol containing β-mercaptoethanol resulted in cells that expressed significant amounts of intracellular YKL-40 protein that was not secreted, which is not seen in normal cells. Thus the synthesis of YKL-40 protein is a marker for MSC differentiation into mature mesenchymal phenotypes, and the presence of untranslated YKL-40 mRNA in non-mesenchymal cells derived from MSCs reflects differences between differentiated and

  15. Purification and characterization of two bifunctional chitinases/lysozymes extracellularly produced by Pseudomonas aeruginosa K-187 in a shrimp and crab shell powder medium.

    OpenAIRE

    Wang, S.L.; Chang, W T

    1997-01-01

    Two extracellular chitinases (FI and FII) were purified from the culture supernatant of Pseudomonas aeruginosa K-187. The molecular weights of FI and FII were 30,000 and 32,000, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 60,000 and 30,000, respectively, by gel filtration. The pIs for FI and FII were 5.2 and 4.8, respectively. The optimum pH, optimum temperature, pH stability, and thermal stability of FI were pH 8, 50 degrees C, pH 6 to 9, and 50 degrees C; ...

  16. Predicting detection performance with model observers: Fourier domain or spatial domain?

    Science.gov (United States)

    Chen, Baiyu; Yu, Lifeng; Leng, Shuai; Kofler, James; Favazza, Christopher; Vrieze, Thomas; McCollough, Cynthia

    2016-03-01

    The use of Fourier domain model observer is challenged by iterative reconstruction (IR), because IR algorithms are nonlinear and IR images have noise texture different from that of FBP. A modified Fourier domain model observer, which incorporates nonlinear noise and resolution properties, has been proposed for IR and needs to be validated with human detection performance. On the other hand, the spatial domain model observer is theoretically applicable to IR, but more computationally intensive than the Fourier domain method. The purpose of this study is to compare the modified Fourier domain model observer to the spatial domain model observer with both FBP and IR images, using human detection performance as the gold standard. A phantom with inserts of various low contrast levels and sizes was repeatedly scanned 100 times on a third-generation, dual-source CT scanner at 5 dose levels and reconstructed using FBP and IR algorithms. The human detection performance of the inserts was measured via a 2-alternative-forced-choice (2AFC) test. In addition, two model observer performances were calculated, including a Fourier domain non-prewhitening model observer and a spatial domain channelized Hotelling observer. The performance of these two mode observers was compared in terms of how well they correlated with human observer performance. Our results demonstrated that the spatial domain model observer correlated well with human observers across various dose levels, object contrast levels, and object sizes. The Fourier domain observer correlated well with human observers using FBP images, but overestimated the detection performance using IR images.

  17. Roles for retrotransposon insertions in human disease

    OpenAIRE

    Dustin C Hancks; Kazazian, Haig H.

    2016-01-01

    Over evolutionary time, the dynamic nature of a genome is driven, in part, by the activity of transposable elements (TE) such as retrotransposons. On a shorter time scale it has been established that new TE insertions can result in single-gene disease in an individual. In humans, the non-LTR retrotransposon Long INterspersed Element-1 (LINE-1 or L1) is the only active autonomous TE. In addition to mobilizing its own RNA to new genomic locations via a “copy-and-paste” mechanism, LINE-1 is able...

  18. Penetration of a Single Domain of Bacillus thuringiensis Cry1Ie-Domain I to a Lipid Membrane In vitro

    Institute of Scientific and Technical Information of China (English)

    GUO Shu-yuan; LI Jie; CHEN Zhen; HE Kang-lai

    2014-01-01

    Domain I of the activated Crystal protein from Bacillus thuringiensis has a sevenα-helix bundle structure, which is responsible for membrane channel formation in its insecticidal mechanism. Cry1Ie is toxic to Asian corn borer, Ostrinia furnacalis (Guenée), and plays important roles in insect biological control. The domain I from Cry1Ie has been expressed and puriifed in its normal conformation, as embedded in the full length homologous toxin structure. The membrane insertion ability of this single domain was compared with the full length homologous toxin using a monolayer insertion experiment. The results indicated that the Cry1Ie-domain I had the ability to insert into the lipid monolayer, and this ability is greater than that of the IE648 toxin. However, the state of insertion is not stable and remains for only a short period of time. The Cry1Ie-domain I plays no role in receptor binding as it had a nonspeciifc binding with the brush border membrane vesicles of the Asian corn borer.

  19. Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) Furnace Development

    Science.gov (United States)

    Crouch, Myscha R.; Carswell, William E.; Farmer, Jeff; Rose, Fred; Tidwell, Paul H., II

    2000-01-01

    The Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) are microgravity furnaces under development at Marshall Space Flight Center. The furnaces are being developed for the first Materials Science Research Rack (MSRR-1) of the Materials Science Research Facility (MSRF), one of the first International Space Station (ISS) scientific payloads. QMI is a Bridgman furnace with quench capability for studying interface behavior during directional solidification of metallic and alloy materials. DMI will be a Bridgman-Stockbarger furnace to study diffusion processes in semiconductors. The design for each insert, both QMI and DMI, is driven by specific science, operations and safety requirements, as well as by constraints arising from resource limitations, such as volume, mass and power. Preliminary QMI analysis and testing indicates that the design meets these requirements.

  20. Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) furnace development

    International Nuclear Information System (INIS)

    The Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) are microgravity furnaces under development at Marshall Space Flight Center. The furnaces are being developed for the first Materials Science Research Rack (MSRR-1) of the Materials Science Research Facility (MSRF), one of the first International Space Station (ISS) scientific payloads. QMI is a Bridgman furnace with quench capability for studying interface behavior during directional solidification of metallic and alloy materials. DMI will be a Bridgman-Stockbarger furnace to study diffusion processes in semiconductors. The design for each insert, both QMI and DMI, is driven by specific science, operations and safety requirements, as well as by constraints arising from resource limitations, such as volume, mass and power. Preliminary QMI analysis and testing indicates that the design meets these requirements

  1. Insertion and deletion processes in recent human history.

    Directory of Open Access Journals (Sweden)

    Per Sjödin

    Full Text Available BACKGROUND: Although insertions and deletions (indels account for a sizable portion of genetic changes within and among species, they have received little attention because they are difficult to type, are alignment dependent and their underlying mutational process is poorly understood. A fundamental question in this respect is whether insertions and deletions are governed by similar or different processes and, if so, what these differences are. METHODOLOGY/PRINCIPAL FINDINGS: We use published resequencing data from Seattle SNPs and NIEHS human polymorphism databases to construct a genomewide data set of short polymorphic insertions and deletions in the human genome (n = 6228. We contrast these patterns of polymorphism with insertions and deletions fixed in the same regions since the divergence of human and chimpanzee (n = 10,546. The macaque genome is used to resolve all indels into insertions and deletions. We find that the ratio of deletions to insertions is greater within humans than between human and chimpanzee. Deletions segregate at lower frequency in humans, providing evidence for deletions being under stronger purifying selection than insertions. The insertion and deletion rates correlate with several genomic features and we find evidence that both insertions and deletions are associated with point mutations. Finally, we find no evidence for a direct effect of the local recombination rate on the insertion and deletion rate. CONCLUSIONS/SIGNIFICANCE: Our data strongly suggest that deletions are more deleterious than insertions but that insertions and deletions are otherwise generally governed by the same genomic factors.

  2. Human YKL39 (chitinase 3-like protein 2), an osteoarthritis-associated gene, enhances proliferation and type II collagen expression in ATDC5 cells

    International Nuclear Information System (INIS)

    Highlights: ► hYKL-39 expression is increased in osteoarthritic articular chondrocytes. ► To examine the molecular functions of hYKL-39 in chondrocytes, we overexpressed hYKL-39 in chondrocytic ATDC5 cells. ► hYKL-39 enhanced proliferation and colony formation in ATDC5 cells. ► hYKL-39 increased type II collagen expression in ATDC5 cells treated with chondrogenic medium. -- Abstract: Human YKL39 (chitinase 3-like protein 2/CHI3L2) is a secreted 39 kDa protein produced by articular chondrocytes and synoviocytes. Recent studies showed that hYKL-39 expression is increased in osteoarthritic articular chondrocytes suggesting the involvement of hYKL-39 in the progression of osteoarthritis (OA). However little is known regarding the molecular function of hYKL-39 in joint homeostasis. Sequence analyses indicated that hYKL-39 has significant identity with the human chitotorisidase family molecules, although it is considered that hYKL-39 has no enzymatic activity since it lacks putative chitinase catalytic motif. In this study, to examine the molecular function of hYKL-39 in chondrocytes, we overexpressed hYKL-39 in ATDC5 cells. Here we report that hYKL-39 enhances colony forming activity, cell proliferation, and type II collagen expression in these cells. These data suggest that hYKL-39 is a novel growth and differentiation factor involved in cartilage homeostasis

  3. Co-expression of a modified maize ribosome-inactivating protein and a rice basic chitinase gene in transgenic rice plants confers enhanced resistance to sheath blight.

    Science.gov (United States)

    Kim, Ju-Kon; Jang, In-Cheol; Wu, Ray; Zuo, Wei-Neng; Boston, Rebecca S; Lee, Yong-Hwan; Ahn, Il-Pyung; Nahm, Baek Hie

    2003-08-01

    Chitinases, beta-1,3-glucanases, and ribosome-inactivating proteins are reported to have antifungal activity in plants. With the aim of producing fungus-resistant transgenic plants, we co-expressed a modified maize ribosome-inactivating protein gene, MOD1, and a rice basic chitinase gene, RCH10, in transgenic rice plants. A construct containing MOD1 and RCH10 under the control of the rice rbcS and Act1 promoters, respectively, was co-transformed with a plasmid containing the herbicide-resistance gene bar as a selection marker into rice by particle bombardment. Several transformants analyzed by genomic Southern-blot hybridization demonstrated integration of multiple copies of the foreign gene into rice chromosomes. Immunoblot experiments showed that MOD1 formed approximately 0.5% of the total soluble protein in transgenic leaves. RCH10 expression was examined using the native polyacrylamide-overlay gel method, and high RCH10 activity was observed in leaf tissues where endogenous RCH10 is not expressed. R1 plants were analyzed in a similar way, and the Southern-blot patterns and levels of transgene expression remained the same as in the parental line. Analysis of the response of R2 plants to three fungal pathogens of rice, Rhizoctonia solani, Bipolaris oryzae, and Magnaporthe grisea, indicated statistically significant symptom reduction only in the case of R. solani (sheath blight). The increased resistance co-segregated with herbicide tolerance, reflecting a correlation between the resistance phenotype and transgene expression. PMID:12885168

  4. Construction of small-insert and large-insert metagenomic libraries.

    Science.gov (United States)

    Simon, Carola; Daniel, Rolf

    2010-01-01

    The vast majority of the Earth's biological diversity is hidden in uncultured and yet uncharacterized microbial genomes. The construction of metagenomic libraries is a cultivation-independent molecular approach to assess this unexplored genetic reservoir. In the last few years, a high number of novel biocatalysts have been identified by function-based or sequence-based screening of metagenomic libraries. Here, we describe detailed protocols for the construction of metagenomic small-insert and large-insert libraries in plasmids and fosmids, respectively, from environmental DNA. PMID:20830554

  5. Polymeric Biodegradable Stent Insertion in the Esophagus

    Directory of Open Access Journals (Sweden)

    Kai Yang

    2016-04-01

    Full Text Available Esophageal stent insertion has been used as a well-accepted and effective alternative to manage and improve the quality of life for patients diagnosed with esophageal diseases and disorders. Current stents are either permanent or temporary and are fabricated from either metal or plastic. The partially covered self-expanding metal stent (SEMS has a firm anchoring effect and prevent stent migration, however, the hyperplastic tissue reaction cause stent restenosis and make it difficult to remove. A fully covered SEMS and self-expanding plastic stent (SEPS reduced reactive hyperplasia but has a high migration rate. The main advantage that polymeric biodegradable stents (BDSs have over metal or plastic stents is that removal is not require and reduce the need for repeated stent insertion. But the slightly lower radial force of BDS may be its main shortcoming and a post-implant problem. Thus, strengthening support of BDS is a content of the research in the future. BDSs are often temporarily effective in esophageal stricture to relieve dysphagia. In the future, it can be expect that biodegradable drug-eluting stents (DES will be available to treat benign esophageal stricture, perforations or leaks with additional use as palliative modalities for treating malignant esophageal stricture, as the bridge to surgery or to maintain luminal patency during neoadjuvant chemoradiation.

  6. Horizontally separated 1-in-1 crossing insertions

    Energy Technology Data Exchange (ETDEWEB)

    Syphers, M.J. [Fermi National Accelerator Lab., Batavia, IL (United States)

    1985-10-01

    Previous to this workshop, realistic lattices have been developed for vertically separated l-in-l (e.g., D.E. Johnson, A.A. Garren) and 2-in-1 (e.g., S. Heifets) magnets as well as for horizontally separated 2-in-l magnets (e.g., SSC RDS). Bringing together the widely separated ({approximately}60-70 cm) beams in a reasonable length of tunnel and keeping the dispersion zero at the interaction point has been difficult in the vertical l-in-l case. Most designs have required spacial 2-in-1 quadrupoles near the interaction point where the beams are separated by 15 cm or less. It is not clear that such magnets, as dictated by some of these lattice designs, can easily be built. The purpose of this exercise is to provide a crossing insertion for a realistic lattice which involves horizontally separated l-in-l magnets. The following horizontal crossing insertions, which incorporate the dispersion suppressors and phase trombones into the major arcs, need no special 2-in-1 magnets near the interaction point. The dispersion at the IP created by the horizontal crossing can be cancelled by the dispersion suppressor and one set of triplets.

  7. Membrane and Protein Interactions of the Pleckstrin Homology Domain Superfamily.

    Science.gov (United States)

    Lenoir, Marc; Kufareva, Irina; Abagyan, Ruben; Overduin, Michael

    2015-01-01

    The human genome encodes about 285 proteins that contain at least one annotated pleckstrin homology (PH) domain. As the first phosphoinositide binding module domain to be discovered, the PH domain recruits diverse protein architectures to cellular membranes. PH domains constitute one of the largest protein superfamilies, and have diverged to regulate many different signaling proteins and modules such as Dbl homology (DH) and Tec homology (TH) domains. The ligands of approximately 70 PH domains have been validated by binding assays and complexed structures, allowing meaningful extrapolation across the entire superfamily. Here the Membrane Optimal Docking Area (MODA) program is used at a genome-wide level to identify all membrane docking PH structures and map their lipid-binding determinants. In addition to the linear sequence motifs which are employed for phosphoinositide recognition, the three dimensional structural features that allow peripheral membrane domains to approach and insert into the bilayer are pinpointed and can be predicted ab initio. The analysis shows that conserved structural surfaces distinguish which PH domains associate with membrane from those that do not. Moreover, the results indicate that lipid-binding PH domains can be classified into different functional subgroups based on the type of membrane insertion elements they project towards the bilayer. PMID:26512702

  8. Membrane and Protein Interactions of the Pleckstrin Homology Domain Superfamily

    Directory of Open Access Journals (Sweden)

    Marc Lenoir

    2015-10-01

    Full Text Available The human genome encodes about 285 proteins that contain at least one annotated pleckstrin homology (PH domain. As the first phosphoinositide binding module domain to be discovered, the PH domain recruits diverse protein architectures to cellular membranes. PH domains constitute one of the largest protein superfamilies, and have diverged to regulate many different signaling proteins and modules such as Dbl homology (DH and Tec homology (TH domains. The ligands of approximately 70 PH domains have been validated by binding assays and complexed structures, allowing meaningful extrapolation across the entire superfamily. Here the Membrane Optimal Docking Area (MODA program is used at a genome-wide level to identify all membrane docking PH structures and map their lipid-binding determinants. In addition to the linear sequence motifs which are employed for phosphoinositide recognition, the three dimensional structural features that allow peripheral membrane domains to approach and insert into the bilayer are pinpointed and can be predicted ab initio. The analysis shows that conserved structural surfaces distinguish which PH domains associate with membrane from those that do not. Moreover, the results indicate that lipid-binding PH domains can be classified into different functional subgroups based on the type of membrane insertion elements they project towards the bilayer.

  9. Superconducting insertion devices with switchable period length

    International Nuclear Information System (INIS)

    Superconducting insertion devices (IDs) are very attractive for synchrotron light sources since they offer the possibility to enhance the tuning range and functionality significantly by period length switching. Period length switching can be realized by employing two or more individually powerable subsets of superconducting coils and reverse the current in a part of the winding. So far, the first demonstration mock-up coil allowing period length tripling was fabricated and tested successfully (A. Grau et al., accepted for publication in IEEE Transactions on Applied Superconductivity). Here, we report on the feasibility of a superconducting switch implemented as proposed by A. Madur et al., Proc. of the 10th International Conference on Synchrotron Radiation Instrumentation 2009, Melbourne, Australia.

  10. Superconducting insertion devices with switchable period length

    Energy Technology Data Exchange (ETDEWEB)

    Holubek, Tomas; Baumbach, Tilo; Casalbuoni, Sara; Gerstl, Stefan; Grau, Andreas; Hagelstein, Michael; Saez de Jauregui, David [Karlsruhe Institute of Technology (KIT), Karlsruhe (Germany); Boffo, Cristian; Walter, Wolfgang [Babcock Noell GmbH, Wuerzburg (Germany)

    2011-07-01

    Superconducting insertion devices (IDs) are very attractive for synchrotron light sources since they offer the possibility to enhance the tuning range and functionality significantly by period length switching. Period length switching can be realized by employing two or more individually powerable subsets of superconducting coils and reverse the current in a part of the winding. So far, the first demonstration mock-up coil allowing period length tripling was fabricated and tested successfully (A. Grau et al., accepted for publication in IEEE Transactions on Applied Superconductivity). Here, we report on the feasibility of a superconducting switch implemented as proposed by A. Madur et al., Proc. of the 10th International Conference on Synchrotron Radiation Instrumentation 2009, Melbourne, Australia.

  11. INSERTION DEVICE UPGRADE PLANS AT THE NSLS.

    Energy Technology Data Exchange (ETDEWEB)

    TANABE, T.; BLEDNYKH, A.; HARDER, D.; LEHECKA, M.; RAKOWSKY, G.; SKARITKA, J.

    2005-05-16

    This paper describes plans to upgrade insertion devices (IDs) at the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, U.S.A. The aging wiggler (W120) at X25 is being replaced by a 1 m long in-vacuum mini-gap undulator (MGU-X25) optimized for a dedicated macromolecular crystallography program. A new, 1/3 m long, undulator (MGU or SCU-X9), will be installed between a pair of RF cavities at X9, and will serve a new beamline dedicated for small angle x-ray scattering (SAXS). Both IDs will have provision for cryocooling the NdFeB hybrid arrays to 150K to raise the field and K-value and to obtain better spectral coverage. Design issues of the devices and other considerations, especially magnetic measurement at low temperature, will be discussed.

  12. Regulatory perspective on incomplete control rod insertions

    Energy Technology Data Exchange (ETDEWEB)

    Chatterton, M.

    1997-01-01

    The incomplete control rod insertions experienced at South Texas Unit 1 and Wolf Creek are of safety concern to the NRC staff because they represent potential precursors to loss of shutdown margin. Even before it was determined if these events were caused by the control rods or by the fuel there was an apparent correlation of the problem with high burnup fuel. It was determined that there was also a correlation between high burnup and high drag forces as well as with rod drop time histories and lack of rod recoil. The NRC staff initial actions were aimed at getting a perspective on the magnitude of the problem as far as the number of plants and the amount of fuel that could be involved, as well as the safety significance in terms of shutdown margin. As tests have been performed and data has been analyzed the focus has shifted more toward understanding the problem and the ways to eliminate it. At this time the staff`s understanding of the phenomena is that it was a combination of factors including burnup, power history and temperature. The problem appears to be very sensitive to these factors, the interaction of which is not clearly understood. The model developed by Westinghouse provides a possible explanation but there is not sufficient data to establish confidence levels and sensitivity studies involving the key parameters have not been done. While several fixes to the problem have been discussed, no definitive fixes have been proposed. Without complete understanding of the phenomena, or fixes that clearly eliminate the problem the safety concern remains. The safety significance depends on the amount of shutdown margin lost due to incomplete insertion of the control rods. Were the control rods to stick high in the core, the reactor could not be shutdown by the control rods and other means such as emergency boration would be required.

  13. Exploring Beta-Amyloid Protein Transmembrane Insertion Behavior and Residue-Specific Lipid Interactions in Lipid Bilayers Using Multiscale MD Simulations

    Science.gov (United States)

    Qiu, Liming; Vaughn, Mark; Cheng, Kelvin

    2013-03-01

    Beta-amyloid (Abeta) interactions with neurons are linked to Alzheimer's. Using a multiscale MD simulation strategy that combines the high efficiency of phase space sampling of coarse-grained MD (CGD) and the high spatial resolution of Atomistic MD (AMD) simulations, we studied the Abeta insertion dynamics in cholesterol-enriched and -depleted lipid bilayers that mimic the neuronal membranes domains. Forward (AMD-CGD) and reverse (CGD-AMD) mappings were used. At the atomistic level, cholesterol promoted insertion of Abeta with high (folded) or low (unfolded) helical contents of the lipid insertion domain (Lys28-Ala42), and the insertions were stabilized by the Lys28 snorkeling and Ala42-anchoring to the polar lipid groups of the bilayer up to 200ns. After the forward mapping, the folded inserted state switched to a new extended inserted state with the Lys28 descended to the middle of the bilayer while the unfolded inserted state migrated to the membrane surface up to 4000ns. The two new states remained stable for 200ns at the atomistic scale after the reverse mapping. Our results suggested that different Abeta membrane-orientation states separated by free energy barriers can be explored by the multiscale MD more effectively than by Atomistic MD simulations alone. NIH RC1-GM090897-02

  14. Protein domain prediction

    NARCIS (Netherlands)

    Ingolfsson, Helgi; Yona, Golan

    2008-01-01

    Domains are considered to be the building blocks of protein structures. A protein can contain a single domain or multiple domains, each one typically associated with a specific function. The combination of domains determines the function of the protein, its subcellular localization and the interacti

  15. Wear mechanisms of WC-Co drill bit inserts against alumina counterface under dry friction: Part 2 — Graded WC-Co inserts

    OpenAIRE

    Yahiaoui, Malik; Paris, Jean-Yves; Denape, Jean; Colin, Christophe; Ther, Olivier; Dourfaye, Alfazazi

    2015-01-01

    The tribological behaviour of innovative graded cemented carbide inserts were studied by using a rotary tribometer and abrasive alumina counterfaces. This work completes the study made on commercial inserts with homogeneous cobalt content. Inserts with three types of graduation processes were considered: inserts with borides WCoB phases, imbibed inserts and inserts combining both processes (i.e. inserts with reactive imbibition). Physicochemical and mechanical measurements show that the WCoB ...

  16. The Effect of Windings on ADSL Transformer Insertion Losses

    Institute of Scientific and Technical Information of China (English)

    JIANG Xiao-na; LAN Zhong-wen; CHEN Sheng-ming; ZHANG Huai-wu; SU Hua

    2007-01-01

    Insertion loss (IL) is one of the important parameters of asymmetrical digital subscriber loop (ADSL) transformers. In different frequency bands, the factors that affect insertion loss are different. Windings mainly affect insertion loss in mid and high frequency bands.The effects of winding ways, winding wire diameter and winding turns on insertion loss were discussed. The presented experiment shows that the insertion loss of an ADSL transformer could be under 0.4 dB in mid frequency band when the winding is 30 turns, in which the ADSL transformer satisfies the requirement of total harmonic distortion (THD). Our experiments also show that the sandwich winding structure is better than the side by side winding structure and the twisted-pair winding structure, and the increase of winding diameter is one means to reduce insertion losses of an ADSL transformer in mid frequency band.

  17. Ocular inserts - Advancement in therapy of eye diseases

    Directory of Open Access Journals (Sweden)

    Anita Kumari

    2010-01-01

    Full Text Available The ocular insert represents a significant advancement in the therapy of eye disease. Ocular inserts are defined as sterile, thin, multilayered, drug-impregnated, solid or semisolid consistency devices placed into the cul-de-sac or conjuctival sac, whose size and shape are especially designed for ophthalmic application. They are composed of a polymeric support that may or may not contain a drug. The drug can later be incorporated as dispersion or a solution in the polymeric support. They offer several advantages as increased ocular residence and sustained release of medication into the eye. The insert includes a body portion sized to position within a lachrymal canaliculus of the eyelid. The inserts are classified according to their solubility as insoluble, soluble, or bioerodible inserts. The release of drug from the insert depends upon the diffusion, osmosis, and bioerosion of the drug, and this article is an attempt to present a brief about this newer drug delivery system.

  18. Insertion torque versus mechanical resistance of mini- implants inserted in different cortical thickness

    Directory of Open Access Journals (Sweden)

    Renata de Faria Santos

    2014-06-01

    Full Text Available OBJECTIVE: This study aimed to measure insertion torque, tip mechanical resistance to fracture and transmucosal neck of mini-implants (MI (Conexão Sistemas de PróteseT, as well as to analyze surface morphology. METHODS: Mechanical tests were carried out to measure the insertion torque of MIs in different cortical thicknesses, and tip mechanical resistance to fracture as well as transmucosal neck of MIs. Surface morphology was assessed by scanning electron microscopy (SEM before and after the mechanical tests. RESULTS: Values of mechanical resistance to fracture (22.14 N.cm and 54.95 N.cm were higher and statistically different (P 0.05 to torsional fracture in the tip of MI (22.14 N.cm when 3 mm cortical thickness (16.11 N.cm and dense bone (23.95 N.cm were used. Torsional fracture of the transmucosal neck (54.95 N.cm was higher and statistically different (P < 0.05 from insertion torsional strength in all tested situations. SEM analysis showed that the MIs had the same smooth surface when received from the manufacturer and after the mechanical tests were performed. Additionally, no significant marks resulting from the manufacturing process were observed. CONCLUSION: All mini-implants tested presented adequate surface morphology. The resistance of mini-implants to fracture safely allows placement in 1 and 2-mm cortical thickness. However, in 3-mm cortical thickness and dense bones, pre-drilling with a bur is recommended before insertion.

  19. EXPLORATION ON SCALABILITY OF DATABASE BULK INSERTION WITH MULTITHREADING

    OpenAIRE

    Boon-Wee Low; Boon-Yaik Ooi; and Chee-Siang Wong

    2011-01-01

    The advancement of database engine and multi-core processors technologies have enable database insertion to be implemented concurrently via multithreading programming. The objective of this work is to evaluate the performance of using multithreading technique to perform database insertion of large data set with known size to enhance the performance of data access layer (DAL) particularly on the bulk-insertion operation. The performance evaluation includes techniques such as using single datab...

  20. Helix insertion into bilayers and the evolution of membrane proteins

    OpenAIRE

    Renthal, Robert

    2009-01-01

    Polytopic α-helical membrane proteins cannot spontaneously insert into lipid bilayers without assistance from polytopic α-helical membrane proteins that already reside in the membrane. This raises the question of how these proteins evolved. Our current knowledge of the insertion of α-helices into natural and model membranes is reviewed with the goal of gaining insight into the evolution of membrane proteins. Topics include: translocon-dependent membrane protein insertion, antibiotic peptides ...

  1. RNA-Editing with Combined Insertion and Deletion Preserves Regularity

    OpenAIRE

    Vink, E.P.; Zantema, H.; Bosnacki, D.

    2013-01-01

    We consider two elementary forms of string rewriting called guided insertion/deletion and guided rewriting. The original strings are modified depending on the match with a given set of auxiliary strings, called guides. Guided insertion/deletion considers matching of a string and a guide with respect to a specific correspondence of strings. Guided rewriting considers matching of a string and a guide with respect to an equivalence relation on the alphabet. Guided insertion/deletion is inspired ...

  2. Internal carotid artery rupture caused by carotid shunt insertion

    OpenAIRE

    Giulio Illuminati; Caliò, Francesco G.; Giulia Pizzardi; Francesco Vietri

    2015-01-01

    Introduction: Shunting is a well-accepted method of maintaining cerebral perfusion during carotid endarterectomy (CEA). Nonetheless, shunt insertion may lead to complications including arterial dissection, embolization, and thrombosis. We present a complication of shunt insertion consisting of arterial wall rupture, not reported previously. Presentation of case: A 78-year-old woman underwent CEA combined with coronary artery bypass grafting (CABG). At the time of shunt insertion an arteria...

  3. Technical note: Computer-manufactured inserts for prosthetic sockets.

    Science.gov (United States)

    Sanders, Joan E; McLean, Jake B; Cagle, John C; Gardner, David W; Allyn, Katheryn J

    2016-08-01

    The objective of this research was to use computer-aided design software and a tabletop 3-D additive manufacturing system to design and fabricate custom plastic inserts for trans-tibial prosthesis users. Shape quality of inserts was tested right after they were inserted into participant's test sockets and again after four weeks of wear. Inserts remained properly positioned and intact throughout testing. Right after insertion the inserts caused the socket to be slightly under-sized, by a mean of 0.11mm, approximately 55% of the thickness of a nylon sheath. After four weeks of wear the under-sizing was less, averaging 0.03mm, approximately 15% of the thickness of a nylon sheath. Thus the inserts settled into the sockets over time. If existing prosthetic design software packages were enhanced to conduct insert design and to automatically generate fabrication files for manufacturing, then computer manufactured inserts may offer advantages over traditional methods in terms of speed of fabrication, ease of design, modification, and record keeping. PMID:27212209

  4. Clinical experience with the percutaneously inserted Amplatz vena caval filter

    International Nuclear Information System (INIS)

    The Amplatz filter is a 14-F vena caval filter designed for percutaneous insertion and retrieval. Thirty filters were inserted. The indications for placement were anticoagulant failure (four cases) and contraindication to anticoagulation (26 cases). Radiologic, pathologic, or clinical follow-up (average, 10 months) was available in 27 patients. Results include one case of recurrent pulmonary embolism (4%), nine cases of caval thrombosis (33%), and two cases of deep venous thrombosis in the insertion leg (7%). Filter migration did not occur in any case. The Amplatz filter is easily inserted and well tolerated by the patient. Although effective in preventing recurrent pulmonary embolism, it has a relatively high incidence of caval thrombosis

  5. The ATLAS Insertable B-Layer

    CERN Document Server

    Alkire, S P; The ATLAS collaboration

    2014-01-01

    During the first shutdown of LHC, the ATLAS detector is undergoing an upgrade of the Pixel Detector in preparation for high luminosity LHC. This upgrade consists of a new pixel layer, called the Insertable B-Layer (IBL), which is positioned between the former inner layer of the Pixel Detector (B-Layer) and a new, smaller, beam pipe, at a radial distance of 3.3 cm from the beam axis. The realization of the IBL required the development of several new technologies and solutions in order to overcome the challenges introduced by the extreme environment and working conditions, such as the high radiation levels, the high pixel occupancy and the need of an exceptionally low material budget. Two silicon sensor technologies have been adopted for the IBL modules: planar n-in-n and 3D. Both of these are connected via bump bonding to the new generation 130 nm IBM CMOS FE-I4 front-end read-out chip. An overview of the IBL project, module design, the qualification tests, data acquisition, results and performance is presente...

  6. APS insertion devices: Recent developments and results

    International Nuclear Information System (INIS)

    The Advanced Photon Source (APS) now has a total of 23 insertion devices (IDs). Over two-thirds of them are installed on the storage ring. The installed devices include 18, 27 and 55 mm-period undulators; an 85 mm-period wiggler; a 16 cm-period elliptical multipole wiggler; and many 33 mm-period undulators. Most of the IDs occupy storage-ring straight sections equipped with 8 mm vertical-aperture vacuum chambers. All of the IDs were measured magnetically at the APS and, in most cases, underwent a final magnetic tuning in order to minimize variation in the various integrals of the field through the ID over the full gap range. Special shimming techniques to correct magnetic field parameters in appropriate gap-dependent ways were developed and applied. Measurements of the closed-orbit distortion as a function of the ID gap variation have been completed, and results are in a good agreement with magnetic measurements. Spectral diagnostics of the ID radiation, including measurements of the absolute spectral flux, brilliance and polarization, show excellent agreement between calculated and measured results. Studies of the sensitivity of IDs to radiation exposure and measurements of the dose rate received by the IDs are in progress

  7. Data insertion in volcanic ash cloud forecasting

    Directory of Open Access Journals (Sweden)

    Kate Louise Wilkins

    2015-03-01

    Full Text Available During the eruption of Eyjafjallajökull in April and May 2010, the London Volcanic Ash Advisory Centre demonstrated the importance of infrared (IR satellite imagery for monitoring volcanic ash and validating the Met Office operational model, NAME. This model is used to forecast ash dispersion and forms much of the basis of the advice given to civil aviation. NAME requires a source term describing the properties of the eruption plume at the volcanic source. Elements of the source term are often highly uncertain and significant effort has therefore been invested into the use of satellite observations of ash clouds to constrain them. This paper presents a data insertion method, where satellite observations of downwind ash clouds are used to create effective ‘virtual sources’ far from the vent. Uncertainty in the model output is known to increase over the duration of a model run, as inaccuracies in the source term, meteorological data and the parameterizations of the modelled processes accumulate. This new technique, where the dis- persion model (DM is ‘reinitialized’ part-way through a run, could go some way to addressing this. 

  8. The ATLAS Insertable B-Layer Project

    CERN Document Server

    Miucci, A; The ATLAS collaboration

    2014-01-01

    The ATLAS experiment will upgrade its Pixel Detector with the installation of a new pixel layer in 2013-14. The new sub-detector, named Insertable B-layer (IBL), will be installed between the existing Pixel Detector and a new smaller radius beam-pipe at a radius of 3.3 cm. To cope with the high radiation and pixel occupancy due to the proximity to the interaction point, a new read-out chip and two different silicon sensor technologies (planar and 3D) have been de- veloped. Furthermore, the physics performance should be improved through the reduction of pixel size while a low material budget should be imposed. A new mechanical support using lightweight staves and a CO2 based cooling system is used. An overview of the IBL project and the status of the production of staves and the qualification of the assembly procedure, the loaded module electrical integrity and the read-out chain will be presented.

  9. Adapting Mathematical Domain Reasoners

    CERN Document Server

    Heeren, Bastiaan

    2010-01-01

    Mathematical learning environments help students in mastering mathematical knowledge. Mature environments typically offer thousands of interactive exercises. Providing feedback to students solving interactive exercises requires domain reasoners for doing the exercise-specific calculations. Since a domain reasoner has to solve an exercise in the same way a student should solve it, the structure of domain reasoners should follow the layered structure of the mathematical domains. Furthermore, learners, teachers, and environment builders have different requirements for adapting domain reasoners, such as providing more details, disallowing or enforcing certain solutions, and combining multiple mathematical domains in a new domain. In previous work we have shown how domain reasoners for solving interactive exercises can be expressed in terms of rewrite strategies, rewrite rules, and views. This paper shows how users can adapt and configure such domain reasoners to their own needs. This is achieved by enabling users...

  10. Transplastomic Nicotiana benthamiana plants expressing multiple defence genes encoding protease inhibitors and chitinase display broad-spectrum resistance against insects, pathogens and abiotic stresses.

    Science.gov (United States)

    Chen, Peng-Jen; Senthilkumar, Rajendran; Jane, Wann-Neng; He, Yong; Tian, Zhihong; Yeh, Kai-Wun

    2014-05-01

    Plastid engineering provides several advantages for the next generation of transgenic technology, including the convenient use of transgene stacking and the generation of high expression levels of foreign proteins. With the goal of generating transplastomic plants with multiresistance against both phytopathogens and insects, a construct containing a monocistronic patterned gene stack was transformed into Nicotiana benthamiana plastids harbouring sweet potato sporamin, taro cystatin and chitinase from Paecilomyces javanicus. Transplastomic lines were screened and characterized by Southern/Northern/Western blot analysis for the confirmation of transgene integration and respective expression level. Immunogold localization analyses confirmed the high level of accumulation proteins that were specifically expressed in leaf and root plastids. Subsequent functional bioassays confirmed that the gene stacks conferred a high level of resistance against both insects and phytopathogens. Specifically, larva of Spodoptera litura and Spodoptera exigua either died or exhibited growth retardation after ingesting transplastomic plant leaves. In addition, the inhibitory effects on both leaf spot diseases caused by Alternaria alternata and soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum were markedly observed. Moreover, tolerance to abiotic stresses such as salt/osmotic stress was highly enhanced. The results confirmed that the simultaneous expression of sporamin, cystatin and chitinase conferred a broad spectrum of resistance. Conversely, the expression of single transgenes was not capable of conferring such resistance. To the best of our knowledge, this is the first study to demonstrate an efficacious stacked combination of plastid-expressed defence genes which resulted in an engineered tolerance to various abiotic and biotic stresses. PMID:24479648

  11. A comparative study of clinical outcomes of post placental insertion versus interval insertion of Copper T 380A intrauterine device

    OpenAIRE

    Suchi Gupta; Shubha Sagar Trivedi; Ratna Biswas

    2015-01-01

    Background: The objective of this study was to study the safety and efficacy of PPIUCD insertion and to compare it with interval insertion. Methods: A prospective study was conducted enrolling 100 women as study group (PPIUCD) and 100 as control group (interval CuT insertion). All women were followed up for 6 months and clinical outcomes were measured in terms of safety, efficacy, effect on menstrual cycles and continuation rates. Chi square test was used to detect differences in the rate...

  12. Clinical information in drug package inserts in India

    Directory of Open Access Journals (Sweden)

    Shivkar Y

    2009-01-01

    Full Text Available Background: It is widely recognized that accurate and reliable product information is essential for the safe and effective use of medications. Pharmaceutical companies are the primary source of most drug information, including package inserts. Package inserts are printed leaflets accompanying marketed drug products and contain information approved by the regulatory agencies. Studies on package inserts in India, in 1996, had shown that crucial information was often missing and they lacked uniformity. Aim: To assess the presentation and completeness of clinically important information provided in the currently available package inserts in India. Materials and Methods: Package inserts accompanying allopathic drug products marketed by pharmaceutical companies in India were collected. These package inserts were analyzed for the content of clinically important information in various sections. Statistical Analysis: The results were expressed as absolute numbers and percentages. Results: Preliminary analyses revealed that most package inserts did contain information under headings, such as, therapeutic indications, contraindications, undesirable effects, etc., listed in the Drugs and Cosmetics Rules 1945. The findings indicated considerable improvement in package inserts since 1996. However, on critical evaluation it was revealed that clinically important information was not well presented and was often incomplete. Information with regard to pediatric and geriatric use was present in only 44% and 13% of the package inserts, respectively. Only five of the inserts had information on the most frequent adverse drug reactions associated with the drug. Also, information on interactions and overdosage was often missing. Conclusion: Although the package inserts appear to have improved over the past decade there is still a definite need to further refine the clinical information contained, to minimize the risks to patients. This could be brought about by self

  13. The activation of cellular oncogenes by retroviral insertion

    International Nuclear Information System (INIS)

    Replication-competent retroviruses can induce a variety of tumors by insertional activation of cellular oncogenes. Transposon tagging techniques have uncovered many novel cellular genes implicated in tumorigenesis. Activation of these genes can occur by insertion of viral promoters, transcriptional enhancement over large distances, or the generation of novel chimeric proteins

  14. Load beam unit replaceable inserts for dry coal extrusion pumps

    Science.gov (United States)

    Saunders, Timothy; Brady, John D.

    2012-11-13

    A track assembly for a particulate material extrusion pump according to an exemplary aspect of the present disclosure includes a link assembly with a roller bearing. An insert mounted to a load beam located such that the roller bearing contacts the insert.

  15. 21 CFR 310.515 - Patient package inserts for estrogens.

    Science.gov (United States)

    2010-04-01

    ... recent revision of the patient package insert. (d) Guidance language. The Food and Drug Administration... 21 Food and Drugs 5 2010-04-01 2010-04-01 false Patient package inserts for estrogens. 310.515 Section 310.515 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN...

  16. Enhanced citrate production through gene insertion in Aspergillus niger

    DEFF Research Database (Denmark)

    Jongh, Wian de; Nielsen, Jens

    2007-01-01

    The effect of inserting genes involved in the reductive branch of the tricarboxylic acid (TCA) cycle on citrate production by Aspergillus niger was evaluated. Several different genes were inserted individually and in combination, i.e. malate dehydrogenase (mdh2) from Saccharomyces cerevisiae, two...

  17. Z-2 Threaded Insert Design and Testing Abstract

    Science.gov (United States)

    Rhodes, RIchard; Graziosi, Dave; Jones, Bobby; Ferl, Jinny; Scarborough, Steve; Sweeney, Mitch

    2016-01-01

    The Z-2 Prototype Planetary Extravehicular Space Suit Assembly is a continuation of NASA's Z series of spacesuits. The Z-2 is another step in the NASA's technology development roadmap leading to human exploration of the Martian surface. To meet a more challenging set of requirements than previous suit systems standard design features, such as threaded inserts, have been re-analyzed and improved. NASA's Z-2 prototype space suit contains several components fabricated from an advanced hybrid composite laminate consisting of IM10 carbon fiber and fiber glass. One requirement NASA levied on the suit composites was the ability to have removable, replaceable helicoil inserts to which other suit components would be fastened. An approach utilizing bonded in inserts with helicoils inside of them was implemented. The design of the interface flanges of the composites allowed some of the inserts to be a "T" style insert that was installed through the entire thickness of the laminate. The flange portion of the insert provides a mechanical lock as a redundancy to the adhesive aiding in the pullout load that the insert can withstand. In some locations it was not possible to utilize at "T" style insert and a blind insert was used instead. These inserts rely completely on the bond strength of the adhesive to resist pullout. It was determined during the design of the suit that the inserts did not need to withstand loads induced from pressure cycling but instead tension induced from torqueing the screws to bolt on hardware which creates a much higher stress on them. Bolt tension is determined by dividing the torque on the screw by a k value multiplied by the thread diameter of the bolt. The k value is a factor that accounts for friction in the system. A common value used for k for a non-lubricated screw is 0.2. The k value can go down by as much as 0.1 if the screw is lubricated which means for the same torque, a much larger tension could be placed on the bolt and insert. This paper

  18. Segregation of fluorescent membrane lipids into distinct micrometric domains: evidence for phase compartmentation of natural lipids ?

    OpenAIRE

    D′auria, Ludovic; Van Der Smissen, Patrick; Bruyneel, Frédéric; Courtoy, Pierre J.; Tyteca, Donatienne

    2011-01-01

    Background We recently reported that sphingomyelin (SM) analogs substituted on the alkyl chain by various fluorophores (e.g. BODIPY) readily inserted at trace levels into the plasma membrane of living erythrocytes or CHO cells and spontaneously concentrated into micrometric domains. Despite sharing the same fluorescent ceramide backbone, BODIPY-SM domains segregated from similar domains labelled by BODIPY-D-e-lactosylceramide (D-e-LacCer) and depended on endogenous SM. Methodology/Principal F...

  19. Improving protein crystal quality by selective removal of a Ca2+-dependent membrane-insertion loop

    International Nuclear Information System (INIS)

    Protein engineering dramatically enhances the quality of crystals of a Ca2+-dependent membrane-binding protein. Lipoxygenases (LOXs) catalyze the regiospecific and stereospecific dioxygenation of polyunsaturated membrane-embedded fatty acids. A Ca2+-dependent membrane-binding function was localized to the amino-terminal C2-like domain of 8R-lipoxygenase (8R-LOX) from the soft coral Plexaura homomalla. The 3.2 Å crystal structure of 8R-LOX and spectroscopic data suggested that Ca2+ stabilizes two membrane-insertion loops. Analysis of the protein packing contacts in the crystal lattice indicated that the conformation of one of the two loops complicated efforts to improve the resolution of the X-ray data. A deletion mutant of 8R-LOX in which the corresponding membrane-insertion loop is absent (Δ41–45:GSLOX) was engineered. Removal of the membrane-insertion loop dramatically increases the protein yield from bacterial cultures and the quality of the crystals obtained, resulting in a better than 1 Å improvement in the resolution of the diffraction data

  20. Gas turbine nozzle vane insert and methods of installation

    Science.gov (United States)

    Miller, William John; Predmore, Daniel Ross; Placko, James Michael

    2002-01-01

    A pair of hollow elongated insert bodies are disposed in one or more of the nozzle vane cavities of a nozzle stage of a gas turbine. Each insert body has an outer wall portion with apertures for impingement-cooling of nozzle wall portions in registration with the outer wall portion. The insert bodies are installed into the cavity separately and spreaders flex the bodies toward and to engage standoffs against wall portions of the nozzle whereby the designed impingement gap between the outer wall portions of the insert bodies and the nozzle wall portions is achieved. The spreaders are secured to the inner wall portions of the insert bodies and the bodies are secured to one another and to the nozzle vane by welding or brazing.

  1. Report on converging insert moulding with µ-IM

    DEFF Research Database (Denmark)

    Islam, Aminul

    Task 5.2.1 deals with the technical feasibility of converging the state-of-the-art µ IM process with insert moulding to offer a wide range of multi-material µ components. The main objective of this deliverable is to summarize state-of-the-art information and to make the guideline needed for the...... convergence. In particular the following aspects are summed up in the deliverable:  Need for converging insert moulding with µ-IM  Objectives and expected outcome from task 5.2.1  State-of-the-art micro insert moulding and different scenario of micro insert moulding  Challenges ahead of converging insert......-IM. Besides this fact current deliverable will provide state-of-the-art information for technology convergence and ways to overcome challenges to use convergent technologies in industrial productions....

  2. Definition of Metrics to Evaluate Cochlear Array Insertion Forces Performed with Forceps, Insertion Tool, or Motorized Tool in Temporal Bone Specimens

    OpenAIRE

    Yann Nguyen; Guillaume Kazmitcheff; Daniele De Seta; Mathieu Miroir; Evelyne Ferrary; Olivier Sterkers

    2014-01-01

    Introduction. In order to achieve a minimal trauma to the inner ear structures during array insertion, it would be suitable to control insertion forces. The aim of this work was to compare the insertion forces of an array insertion into anatomical specimens with three different insertion techniques: with forceps, with a commercial tool, and with a motorized tool. Materials and Methods. Temporal bones have been mounted on a 6-axis force sensor to record insertion forces. Each temporal bone has...

  3. Studies on the Screening of Chitinase Producing Strain from Ocean and Its Optimization on Fermentation%海洋产几丁质酶菌株的筛选及发酵条件优化

    Institute of Scientific and Technical Information of China (English)

    张灿; 黄德智; 李丰硕; 王硕; 薛永常

    2012-01-01

    A strain with high productivity of chitinase was screened through transparent circle method from sea mud of Dalian seaside and was identified as species Brevibacterium linens. After medium optimization and orthogonal experiments, which include carbon sources, nitrogen sources, temperature and pH, the chitinase activity was improved. The results showed that the optimized conditions for chitinase production were 1 g/L peptone, 10 g/L Colloidal chitin, the NaCl of 20 g/L, culture at 28 ?and pH 7.0. Under these conditions the chitinase activity can reach 0.508 U/mL,9 times higher than that of par ent strain.%通过透明圈法初筛、DNS复筛,从大连近海海域的海泥中分离出一株高产几丁质酶的菌株,经初步鉴定为扩展短杆菌(Brevibacterium linens).采用不同碳源、氮源、温度、pH值等对培养条件进行了优化及正交试验.结果表明:该菌产酶最适条件是在28℃、pH7.0、蛋白胨1g/L、胶体几丁质10g/L、NaCl 20g/L条件下,几丁质酶活力可达0.508 U/mL,比优化前其产酶活力提高了9倍.

  4. Reactivity insertion assumptions used in safety analysis calculations

    International Nuclear Information System (INIS)

    The report discusses the basis for selection of the trip reactivity insertion curve used by B and W in the safety analysis of its nuclear steam supply systems. The implementation of new three-dimensional calculational techniques and test results from operating plants now allow derivation of reactivity insertion curves that vary about the previously symmetric insertion curve used in the safety analysis. The report is generically applicable to all operating or nearly operating 177-FA (fuel assembly) plants in its evaluation of the impact of these new trip reactivity insertion curves. In conjunction with the TMI-2 FSAR review, the NRC expressed concern over the scram times defined by the Technical Specifications and how they were related to the safety analysis presented in Chapter 15. This concern is discussed in the report, and a proposed Technical Specification change is presented that is responsive to the staff's concern. The control rod drive mechanism (CRDM) is discussed, and the results of on-site tests are presented. Calculated integral rod worth data are compared to measured data. Also discussed are factors that influence the shape of this curve, such as flux redistribution and initial off-set. The trip reactivity insertion curve used in the safety analysis is discussed. This curve is a composite of the rod velocity curve and the integral rod worth curve. The synthesis of two curves to produce the reactivity insertion curve is illustrated. The impact of the resultant trip reactivity insertion curve on the safety analysis is also described

  5. Immediate post-placental IUD insertion: the expulsion problem.

    Science.gov (United States)

    Thiery, M; Van Kets, H; Van der Pas, H

    1985-04-01

    This paper reports an evaluation of immediate post-placental insertion of a non-copper (Lippes Loop D) and several copper-bearing IUD models (TCu200, TCu220C, MLCu375, MLCu250, Nova T-PP, DimélysR). Based on the analysis of a total of 2,646 insertions and 55,794 woman-months of experience, we conclude that placement of an IUD within ten minutes of delivery of the placenta is a valuable alternative to interval insertion, because this method is safe and effective. Effectiveness was significantly lower for the Lippes Loop D than for the T- and ML-IUD models tested, the latter showing roughly comparable pertinent event rates. Pertinent event rates for copper IUDs were influenced by the skill of the operator; age of the recipient only had a significant effect on effectiveness, whereas parity had no significant effect on pertinent event rates. The single and still unsolved problem associated with immediate postpartum insertion is the greater likelihood of expulsion compared with interval insertion, and this hazard is significantly much greater for the Loop than for the copper-bearing devices assessed. The evolution of the expulsion rates shows a constant time-relationship. This pattern makes it obvious why follow-up of recipients, at least during the first trimester following insertion, is mandatory if immediate post-placental IUD insertion is to be optimally effective. PMID:4006467

  6. Knot Insertion Algorithms for ECT B-spline Curves

    Institute of Scientific and Technical Information of China (English)

    SONG Huan-huan; TANG Yue-hong; LI Yu-juan

    2013-01-01

    Knot insertion algorithm is one of the most important technologies of B-spline method. By inserting a knot the local prop-erties of B-spline curve and the control flexibility of its shape can be further improved, also the segmentation of the curve can be re-alized. ECT spline curve is drew by the multi-knots spline curve with associated matrix in ECT spline space;Muehlbach G and Tang Y and many others have deduced the existence and uniqueness of the ECT spline function and developed many of its important properties .This paper mainly focuses on the knot insertion algorithm of ECT B-spline curve.It is the widest popularization of B-spline Behm algorithm and theory. Inspired by the Behm algorithm, in the ECT spline space, structure of generalized Pólya poly-nomials and generalized de Boor Fix dual functional, expressing new control points which are inserted after the knot by linear com-bination of original control vertex the single knot, and there are two cases, one is the single knot, the other is the double knot. Then finally comes the insertion algorithm of ECT spline curve knot. By application of the knot insertion algorithm, this paper also gives out the knot insertion algorithm of four order geometric continuous piecewise polynomial B-spline and algebraic trigonometric spline B-spline, which is consistent with previous results.

  7. 枯草芽孢杆菌SL-13的生防性能及其抗菌几丁质酶特性研究%Biocontrol Efficiency of Bacillus subtilis SL-13 and Characterization of an Antifungal Chitinase

    Institute of Scientific and Technical Information of China (English)

    刘燕; 陶晶; 阎豫君; 李彬; 李晖; 李春

    2011-01-01

    The seed germination and tomato seedling tests showed that Bacillus subtilis SL-13 could promote the sprouting and seedling growth of tomato. The fresh and dry weight of tomato seedlings increased 42.86% and 18.75%, respectively. The control efficacies of the SL-13 to tomato Rhizoctonia rot were 20.65% and 35.23% in the greenhouse and field, respectively. The growth of the plant-pathogenic fungus Rhizoctonia solani was considerably inhibited in the presence of the strain SL-13 culture supernatant. The main antifungal protein was detected to be chitinase through vitro assay. The chitinase was purified with DEAE-Sepharose fast flow ion exchange column chromatography and Sephadex G-75 gel filtration for further characterization. The optimal pH and temperature for the chitinase activity were 7.0 and 50 ℃, respectively. It was demonstrated that the enzyme was stable at pH 5-9 and 40-60 ℃. 70% of the enzyme activity was retained when incubated at 121 ℃ and 0.11 MPa for 20 min, and the enzyme was not sensitive to protease K and ultraviolet radiation. Thus it is suitable for effective biological control in relatively unstable environment.

  8. Development of ITER toroidal field insert. International collaboration with Russia

    International Nuclear Information System (INIS)

    The Central Solenoid (CS) model coil programme was performed since 1992 as one of the projects in the Engineering Design Activity (EDA) of the International Thermonuclear Experimental Reactor(ITER). The CS model coil programme involves a plan to develop the Toroidal Field (TF) insert to demonstrate the conductor performance of ITER TF coils under a magnetic flux density of 13T. The TF insert was fabricated by Russia and tested by Japan under the framework of the ITER-EDA. The TF insert developed a single-layer solenoid with nine turns. It is wound with a cable-in-conduit (CIC) conductor which consists of 1,152 Nb3Sn strands, a thin titanium jacket and a central channel. The outer diameter, height and weight of the TF insert are 1.56 m, 3.2 m and 3.1 ton, respectively. Fabrication of the TF insert was completed in May 2001 at the D.V.Efremov Scientific Research Institute for Electrophysical Apparatus (Efremov institute) in St. Petersburg, Russia. The TF insert was then transported to the Japan Atomic Energy Research Institute (JAERI). Installation of the TF insert to CS model coil test facility was completed in August, 2001. Experiments including the cooldown and warmup processes, were completed in November 2001. The TF insert was charged to 13T with 46 kA without any instability under a back up magnetic field from the CS model coil. This report introduces an overview of the fabrication, installation and experiments for the TF insert conducted under collaboration between Japan and Russia. (author)

  9. Islet amyloid polypeptide inserts into phospholipid monolayers as monomer.

    Science.gov (United States)

    Engel, Maarten F M; Yigittop, HaciAli; Elgersma, Ronald C; Rijkers, Dirk T S; Liskamp, Rob M J; de Kruijff, Ben; Höppener, Jo W M; Antoinette Killian, J

    2006-02-24

    Amyloid deposits in the pancreatic islets of Langerhans are thought to be a main factor responsible for death of the insulin-producing islet beta-cells in type 2 diabetes. It is hypothesized that beta-cell death is related to interaction of the 37 amino acid residue human islet amyloid polypeptide (hIAPP), the major constituent of islet amyloid, with cellular membranes. However, the mechanism of hIAPP-membrane interactions is largely unknown. Here, we study the nature and the molecular details of the initial step of hIAPP-membrane interactions by using the monolayer technique. It is shown that both freshly dissolved hIAPP and the non-amyloidogenic mouse IAPP (mIAPP) have a pronounced ability to insert into phospholipid monolayers, even at lipid packing conditions that exceed the conditions that occur in biological membranes. In contrast, the fibrillar form of hIAPP has lost the ability to insert. These results, combined with the observations that both the insertion kinetics and the dependence of insertion on the initial surface pressure are similar for freshly dissolved hIAPP and mIAPP, indicate that hIAPP inserts into phospholipid monolayers most likely as a monomer. In addition, our results suggest that the N-terminal part of hIAPP, which is nearly identical with that of mIAPP, is largely responsible for insertion. This is supported by experiments with hIAPP fragments, which show that a peptide consisting of the 19 N-terminal residues of hIAPP efficiently inserts into phospholipid monolayers, whereas an amyloidogenic decapeptide, consisting of residues 20-29 of hIAPP, inserts much less efficiently. The results obtained here suggest that hIAPP monomers might insert with high efficiency in biological membranes in vivo. This process could play an important role as a first step in hIAPP-induced membrane damage in type 2 diabetes. PMID:16403520

  10. Study on Visually Design of Indexable Insert Tip

    Institute of Scientific and Technical Information of China (English)

    YAO Ji-quan; ZHAO Shu-qiang; ZHANG Shen-hou; CHEN Xin; GUO Xian-yang

    2013-01-01

    In order to improve the development efficiency of the indexable insert tip, three-dimensional parametric and visualization applications program of indexable insert tip was developed in AutoCAD VBA (Visual Basic for Applications). The software could significantly improve the modeling efficiency of indexable insert tip and the model could also be used to static analysis, dynamic analysis and thermodynamic analysis. All aspects of performance could be fully understood by the finite element analysis. The ref-erence for the blade parameters preferred is also provided.

  11. Cutting inserts effect on heat generation in turning process

    Directory of Open Access Journals (Sweden)

    Ján Žitňanský

    2014-03-01

    Full Text Available The paper is focused on confirmation of an effect of cutting materials and cutting inserts geometry on cutting forces and temperature, quality and accuracy of machined surface in turning. The main part is devoted to description of advanced turning tools and cutting materials. The experimental part is focused on cutting forces and temperature measurement and machined surface quality in turning of steel grade 11 523 samples with one feed and depth of cut value and varying spindle speed. Measurements were performed with various types of indexable cutting inserts used. Measured values were evaluated in terms of the effect of different properties of cutting inserts and various spindle speed values.

  12. Diamagnetic (Condon) domains

    International Nuclear Information System (INIS)

    This paper is the first systematic review of experimental research on diamagnetic (aka Condon) domains that form in nonmagnetic metals at low temperatures due to the development of Landau levels. A variety of methods were used to study the domains. Muon spectroscopy studies showed such domains to be present in all metals studied, pointing to the universal nature of the phenomenon. For silver, the domain structure size as measured by Hall microsensors turned out to be an order of magnitude larger than expected. In beryllium, it was found that domains do not come to the surface but rather remain in the bulk of the crystal. The magnetostriction of beryllium during domain formation is measured. It is shown that magnetization current in a domain wall is entirely caused by the charge density gradient in the wall, due to the lattice being deformed oppositely in neighboring domains. It is observed for the first time that the de Haas-van Alphen effect exhibits hysteresis at the transition to the domain state, and this fact was used for the experimental determination of the phase diagrams for the domain states of silver and beryllium. (reviews of topical problems)

  13. The effect of ventilation tube insertion or trans-tympanic silicone plug insertion on a patulous Eustachian tube.

    Science.gov (United States)

    Endo, Shiori; Mizuta, Kunihiro; Takahashi, Goro; Nakanishi, Hiroshi; Yamatodani, Takashi; Misawa, Kiyoshi; Hosokawa, Seiji; Mineta, Hiroyuki

    2016-06-01

    Conclusions This study suggests that long-term ventilation tube insertion is the first-choice surgical treatment for a 'sniff-type' patulous Eustachian tube (PET). When treating a refractory PET, it is important to determine whether the patient had a habitual sniff. Objectives PET patients were divided into two groups: patients with a habitual sniff (sniff-type PET) and those without a habitual sniff (non-sniff-type PET). This study examined the effects of ventilation tube insertion or silicone plug insertion in each group. Methods Surgical procedures such as ventilation tube insertion or trans-tympanic silicone plug insertion were performed for these patients. Tubotympanoaero-dynamic graphy (TTAG) was also performed to determine the mechanisms underlying these treatments. Results There were 11 cases (17 ears) of sniff-type PET and 20 cases (27 ears) of non-sniff-type PET. An improvement in symptoms was found in 72.7% of the patients who underwent silicone plug insertion (66.7% for sniff-type PET and 74.1% for non-sniff-type PET) and in 90.9% of the patients who underwent ventilation tube insertion for sniff-type PET. In TTAG assessments, many sniff-type PET patients showed significant synchronous changes at high levels of pressure (over 40 daPa) in the external auditory meatus and nasopharynx when performing a slight Valsalva manoeuvre (below 200 daPa). PMID:26903085

  14. Frustratingly Easy Domain Adaptation

    CERN Document Server

    Daumé, Hal

    2009-01-01

    We describe an approach to domain adaptation that is appropriate exactly in the case when one has enough ``target'' data to do slightly better than just using only ``source'' data. Our approach is incredibly simple, easy to implement as a preprocessing step (10 lines of Perl!) and outperforms state-of-the-art approaches on a range of datasets. Moreover, it is trivially extended to a multi-domain adaptation problem, where one has data from a variety of different domains.

  15. CULTURAL DOMAINS: TRANSLATION PROBLEMS

    OpenAIRE

    Isabel Negro Alousque

    2009-01-01

    Much of the lexis of a language has a cultural referent and is thus specific to a speech community. The meaning of culturally marked words is often difficult to grasp without some cultural knowledge and poses translation problems, particularly when the words are associated with cultural domains (Nida, 2001). In the present paper we focus on the French cultural domain of cooking. After outlining the elements of the domain, we analyse the difficulties in translating the lexical units from the d...

  16. Visualizing latent domain knowledge

    OpenAIRE

    Chen, C.; Kuljis, J; Paul, RJ

    2001-01-01

    Knowledge discovery and data mining commonly rely on finding salient patterns of association from a vast amount of data. Traditional citation analysis of scientific literature draws insights from strong citation patterns. Latent domain knowledge, in contrast to the mainstream domain knowledge, often consists of highly relevant but relatively infrequently cited scientific works. Visualizing latent domain knowledge presents a significant challenge to knowledge discovery and quantitative studies...

  17. Single-copy insertion of transgenes in Caenorhabditis elegans

    DEFF Research Database (Denmark)

    Frøkjaer-Jensen, Christian; Davis, M Wayne; Hopkins, Christopher E; Newman, Blake J; Thummel, Jason M; Olesen, Søren-Peter; Grunnet, Morten; Jorgensen, Erik M

    2008-01-01

    At present, transgenes in Caenorhabditis elegans are generated by injecting DNA into the germline. The DNA assembles into a semistable extrachromosomal array composed of many copies of injected DNA. These transgenes are typically overexpressed in somatic cells and silenced in the germline. We have...... developed a method that inserts a single copy of a transgene into a defined site. Mobilization of a Mos1 transposon generates a double-strand break in noncoding DNA. The break is repaired by copying DNA from an extrachromosomal template into the chromosomal site. Homozygous single-copy insertions can be...... obtained in less than 2 weeks by injecting approximately 20 worms. We have successfully inserted transgenes as long as 9 kb and verified that single copies are inserted at the targeted site. Single-copy transgenes are expressed at endogenous levels and can be expressed in the female and male germlines....

  18. A new insertion sequence for incremental Delaunay triangulation

    Institute of Scientific and Technical Information of China (English)

    Jian-Fei Liu; Jin-Hui Yan; S.H.Lo

    2013-01-01

    Incremental algorithm is one of the most popular procedures for constructing Delaunay triangulations (DTs).However,the point insertion sequence has a great impact on the amount of work needed for the construction of DTs.It affects the time for both point location and structure update,and hence the overall computational time of the triangulation algorithm.In this paper,a simple deterministic insertion sequence is proposed based on the breadth-first-search on a Kd-tree with some minor modifications for better performance.Using parent nodes as search-hints,the proposed insertion sequence proves to be faster and more stable than the Hilbert curve order and biased randomized insertion order (BRIO),especially for non-uniform point distributions over a wide range of benchmark examples.

  19. Insertion device magnet measurements for the Advanced Light Source

    International Nuclear Information System (INIS)

    Allowable magnetic field errors for the 4.6 m long insertion devices for the Advanced Light Source (ALS) are extremely small and are driven by electron beam and radiation requirements. Detailed measurements and adjustments of each insertion device are performed to qualify them for installation in the ALS. To accomplish this, a high speed, precision magnetic measurement facility has been designed and built. Hall probe mapping equipment, capable of completing a 2500 sample, 6 m scan with precision axial position monitoring using a laser interferometer in under one minute, is used to obtain both local and integrated field information. A 5.5 m long, 1 cm wide coil is used to measure the field integral through an entire insertion device. This paper describes magnetic measurement equipment, and results of measurements on IDA, the first of the ALS insertion devices

  20. Posterior cruciate ligament's tibial insertions: topographic anatomy and morphometric study

    OpenAIRE

    Julio Cesar Gali; Hector Campora de Sousa Oliveira; Bruno Cesar Bracher Lisboa; Bruno DiSerio Dias; Fabiana de Godoy Casimiro; Edie Benedito Caetano

    2013-01-01

    OBJECTIVE: To provide anatomical and morphometric basis of the posterior cruciate ligament's tibial insertions in order to assist the creation of anatomical tibial tunnels, in the ligament surgical reconstruction. MATERIAL AND METHODS: The topographic anatomy and morphometry of the posterior cruciate ligament's anterolateral and posteromedial bundles' tibial insertions were analyzed in 24 anatomical knee pieces. The pieces were photographed by a digital camera and the images obtained wer...

  1. Generating Novel Allelic Variation Through Activator Insertional Mutagenesis in Maize

    OpenAIRE

    Bai, Ling; Singh, Manjit; Pitt, Lauren; Sweeney, Meredith; Brutnell, Thomas P.

    2007-01-01

    The maize transposable element Activator (Ac) has been exploited as an insertional mutagen to disrupt, clone, and characterize genes in a number of plant species. To develop an Ac-based mutagenesis platform for maize, a large-scale mutagenesis was conducted targeting the pink scutellum1 locus. We selected 1092 Ac transposition events from a closely linked donor Ac, resulting in the recovery of 17 novel ps1 alleles. Multiple phenotypic classes were identified corresponding to Ac insertions in ...

  2. Impediments to rapid insertion of innovative displays and peripherals

    Science.gov (United States)

    Nicholson, Gail

    2012-06-01

    In order to optimize system performance and minimize cost for a system to fill capability gaps, an improvement to rapid insertion of innovative display and peripheral technology is required to take advantage of human-machine intersections. Current approaches to testing and integration impedes successful rapid insertion of innovative technology for new systems and incremental upgrades. Considerations to innovative displays and peripherals must occur further to the left of the lifecycle to be successful and key integration areas must be address for success.

  3. Blind bedside insertion of small bowel feeding tubes.

    LENUS (Irish Health Repository)

    Duggan, SN

    2009-12-01

    The use of Naso-Jejunal (NJ) feeding is limited by difficulty in feeding tube placement. Patients have traditionally required transfer to Endoscopy or Radiology for insertion of small bowel feeding tubes, with clear resource implications. We hypothesised that the adoption of a simple bedside procedure would be effective and reduce cost. Clinical nutrition and nurse specialist personnel were trained in the 10\\/10\\/10 method of blind bedside NJ insertion.

  4. Heat Transfer Enhancement by Using Different Types of Inserts

    OpenAIRE

    Tabatabaeikia, S.; Mohammed, H.A.; Nik-Ghazali, N.; Shahizare, B.

    2014-01-01

    Heat transfer enhancement has been always a significantly interesting topic in order to develop high efficient, low cost, light weight, and small heat exchangers. The energy cost and environmental issue are also encouraging researchers to achieve better performance than the existing designs. Two of the most effective ways to achieve higher heat transfer rate in heat exchangers are using different kinds of inserts and modifying the heat exchanger tubes. There are different kinds of inserts emp...

  5. Insertion and Deletion Processes in Recent Human History

    OpenAIRE

    Per Sjödin; Thomas Bataillon; Schierup, Mikkel H.

    2010-01-01

    BACKGROUND: Although insertions and deletions (indels) account for a sizable portion of genetic changes within and among species, they have received little attention because they are difficult to type, are alignment dependent and their underlying mutational process is poorly understood. A fundamental question in this respect is whether insertions and deletions are governed by similar or different processes and, if so, what these differences are. METHODOLOGY/PRINCIPAL FINDINGS: We use publishe...

  6. Effects of Rotational Motion in Robotic Needle Insertion

    OpenAIRE

    Ramezanpour, H.; H. Yousefi; M. Rezaei; M Rostami

    2015-01-01

    Background Robotic needle insertion in biological tissues has been known as one the most applicable procedures in sampling, robotic injection and different medical therapies and operations. Objective In this paper, we would like to investigate the effects of angular velocity in soft tissue insertion procedure by considering force-displacement diagram. Non-homogenous camel liver can be exploited as a tissue sample under standard compression test with Zwick/Roell device employing 1-D axial load...

  7. Needle Insertion with Duty-Cycled Rotation into Multiple Media

    OpenAIRE

    Lehocky, Craig A.; Riviere, Cameron N.

    2012-01-01

    Thin, flexible needles can be steered along nonlinear paths to reach deep anatomical structures within the human body. This study builds upon previous work involving steering of bevel-tipped needles by inserting while rotating in a duty-cycled fashion. Here we investigate how needle material and radius, duty cycle, and tissue stiffness affect needle curvature. Needles were inserted into media while rotated at a specified duty cycle and the curvature was measured. A linear relationship between...

  8. Lithium Insertion In Silicon Nanowires: An ab Initio Study

    KAUST Repository

    Zhang, Qianfan

    2010-09-08

    The ultrahigh specific lithium ion storage capacity of Si nanowires (SiNWs) has been demonstrated recently and has opened up exciting opportunities for energy storage. However, a systematic theoretical study on lithium insertion in SiNWs remains a challenge, and as a result, understanding of the fundamental interaction and microscopic dynamics during lithium insertion is still lacking. This paper focuses on the study of single Li atom insertion into SiNWs with different sizes and axis orientations by using full ab initio calculations. We show that the binding energy of interstitial Li increases as the SiNW diameter grows. The binding energies at different insertion sites, which can be classified as surface, intermediate, and core sites, are quite different. We find that surface sites are energetically the most favorable insertion positions and that intermediate sites are the most unfavorable insertion positions. Compared with the other growth directions, the [110] SiNWs with different diameters always present the highest binding energies on various insertion locations, which indicates that [110] SiNWs are more favorable by Li doping. Furthermore, we study Li diffusion inside SiNWs. The results show that the Li surface diffusion has a much higher chance to occur than the surface to core diffusion, which is consistent with the experimental observation that the Li insertion in SiNWs is layer by layer from surface to inner region. After overcoming a large barrier crossing surface-to-intermediate region, the diffusion toward center has a higher possibility to occur than the inverse process. © 2010 American Chemical Society.

  9. Hardening device, by inserts, of electronic component against radiation

    International Nuclear Information System (INIS)

    The hardening device includes at least two materials, one with high atomic number with respect to the other. One of these materials is set as inserts in a layer of the other material. The hardening device is then made by stacking of such layers, the insert density varying from one layer to the other, making thus vary the atomic number resulting from the hardening device along its thickness, following a predefined law

  10. Insertion Loss Analysis of the Acoustic Panels with Composite Construction

    Directory of Open Access Journals (Sweden)

    Vasile Ovidiu

    2013-09-01

    Full Text Available In order to reduce noise pollution, under European legislation in our country need to align, a special attention has the tests carried out on acoustic panels. The most eloquent attempts, defining traffic noise attenuation are tests that can determine the insertion loss of an acoustic screen. In this paper, it briefly shows the method of determining the insertion loss in octave bands and some real results in the case of acoustic panel in a composite construction.

  11. Modeling Amyloid Beta Peptide Insertion into Lipid Bilayers

    CERN Document Server

    Mobley, D L; Singh, R R P; Maddox, M W; Longo, M J; Mobley, David L.; Cox, Daniel L.; Singh, Rajiv R. P.; Maddox, Michael W.; Longo, Marjorie L.

    2003-01-01

    Inspired by recent suggestions that the Alzheimer's amyloid beta peptide (A-beta), can insert into cell membranes and form harmful ion channels, we model insertion of the peptide into cell membranes using a Monte Carlo code which is specific at the amino acid level. We examine insertion of the regular A-beta peptide as well as mutants causing familial Alzheimer's disease. We present our results and develop the hypothesis that partial insertion into the membrane, leaving the peptide in one leaflet, increases the probability of harmful channel formation. This hypothesis can partly explain why these mutations are neurotoxic simply due to peptide insertion behavior, and also explains why, normally, A-beta 42 is more toxic to some cultured cells than A-beta 40, but the E22Q mutation reverses this effect. We further apply this model to various artificial A-beta mutants which have been examined experimentally, and offer testable experimental predictions contrasting the roles of aggregation and insertion with regard ...

  12. Evaluation of a high resolution silicon PET insert module

    Energy Technology Data Exchange (ETDEWEB)

    Grkovski, Milan, E-mail: milan.grkovski@ijs.si [Jožef Stefan Institute, Ljubljana (Slovenia); Memorial Sloan Kettering Cancer Center, New York, NY (United States); Brzezinski, Karol [IFIC/CSIC, Valencia (Spain); Cindro, Vladimir [Jožef Stefan Institute, Ljubljana (Slovenia); Clinthorne, Neal H. [University of Michigan, Ann Arbor, MI (United States); Kagan, Harris [Ohio State University, Columbus, OH (United States); Lacasta, Carlos [IFIC/CSIC, Valencia (Spain); Mikuž, Marko [Jožef Stefan Institute, Ljubljana (Slovenia); Solaz, Carles [IFIC/CSIC, Valencia (Spain); Studen, Andrej [Jožef Stefan Institute, Ljubljana (Slovenia); Weilhammer, Peter [Ohio State University, Columbus, OH (United States); Žontar, Dejan [Jožef Stefan Institute, Ljubljana (Slovenia)

    2015-07-11

    Conventional PET systems can be augmented with additional detectors placed in close proximity of the region of interest. We developed a high resolution PET insert module to evaluate the added benefit of such a combination. The insert module consists of two back-to-back 1 mm thick silicon sensors, each segmented into 1040 1 mm{sup 2} pads arranged in a 40 by 26 array. A set of 16 VATAGP7.1 ASICs and a custom assembled data acquisition board were used to read out the signal from the insert module. Data were acquired in slice (2D) geometry with a Jaszczak phantom (rod diameters of 1.2–4.8 mm) filled with {sup 18}F-FDG and the images were reconstructed with ML-EM method. Both data with full and limited angular coverage from the insert module were considered and three types of coincidence events were combined. The ratio of high-resolution data that substantially improves quality of the reconstructed image for the region near the surface of the insert module was estimated to be about 4%. Results from our previous studies suggest that such ratio could be achieved at a moderate technological expense by using an equivalent of two insert modules (an effective sensor thickness of 4 mm)

  13. RNA-Editing with Combined Insertion and Deletion Preserves Regularity

    Directory of Open Access Journals (Sweden)

    E.P. de Vink

    2013-01-01

    Full Text Available We consider two elementary forms of string rewriting called guided insertion/deletion and guided rewriting. The original strings are modified depending on the match with a given set of auxiliary strings, called guides. Guided insertion/deletion considers matching of a string and a guide with respect to a specific correspondence of strings. Guided rewriting considers matching of a string and a guide with respect to an equivalence relation on the alphabet. Guided insertion/deletion is inspired by {RNA}-editing, a biological process by which the original genetic information stored in DNA is modified before its final expression. The formalism here allows for simultaneous insertion and deletion of string elements. Guided rewriting, based on a letter-to-letter relation, is technically more appealing than guided insertion/deletion. We prove that guided rewriting preserves regularity: for every regular language its closure under guided rewriting is regular too. In the proof we will rely on the auxiliary notion of a slice sequence. We establish a correspondence of slice sequences and guide rewrite sequences. Because of their left-to-right nature, slice sequences are more convenient to deal with than guided rewrite sequences in the construction of the finite automata that we encounter in the proofs of regularity. Based on the result for guided rewriting we establish that guided insertion/deletion preserves regularity as well.

  14. Evaluation of a high resolution silicon PET insert module

    International Nuclear Information System (INIS)

    Conventional PET systems can be augmented with additional detectors placed in close proximity of the region of interest. We developed a high resolution PET insert module to evaluate the added benefit of such a combination. The insert module consists of two back-to-back 1 mm thick silicon sensors, each segmented into 1040 1 mm2 pads arranged in a 40 by 26 array. A set of 16 VATAGP7.1 ASICs and a custom assembled data acquisition board were used to read out the signal from the insert module. Data were acquired in slice (2D) geometry with a Jaszczak phantom (rod diameters of 1.2–4.8 mm) filled with 18F-FDG and the images were reconstructed with ML-EM method. Both data with full and limited angular coverage from the insert module were considered and three types of coincidence events were combined. The ratio of high-resolution data that substantially improves quality of the reconstructed image for the region near the surface of the insert module was estimated to be about 4%. Results from our previous studies suggest that such ratio could be achieved at a moderate technological expense by using an equivalent of two insert modules (an effective sensor thickness of 4 mm)

  15. A new specifically designed forceps for chest drain insertion.

    LENUS (Irish Health Repository)

    Andrews, Emmet

    2012-02-03

    Insertion of a chest drain can be associated with serious complications. It is recommended that the drain is inserted with blunt dissection through the chest wall but there is no specific instrument to aid this task. We describe a new reusable forceps that has been designed specifically to facilitate the insertion of chest drains.A feasibility study of its use in patients who required a chest drain as part of elective cardiothoracic operations was undertaken. The primary end-point was successful and accurate placement of the drain. The operators also completed a questionnaire rating defined aspects of the procedure. The new instrument was used to insert the chest drain in 30 patients (19 male, 11 female; median age 61.5 years (range 16-81 years)). The drain was inserted successfully without the trocar in all cases and there were no complications. Use of the instrument rated as significantly easier relative to experience of previous techniques in all specified aspects. The new device can be used to insert intercostal chest drains safely and efficiently without using the trocar or any other instrument.

  16. Unassisted translocation of large polypeptide domains across phospholipid bilayers

    OpenAIRE

    Brambillasca, Silvia; Yabal, Monica; Makarow, Marja; Borgese, Nica

    2006-01-01

    Although transmembrane proteins generally require membrane-embedded machinery for integration, a few can insert spontaneously into liposomes. Previously, we established that the tail-anchored (TA) protein cytochrome b(5) (b5) can posttranslationally translocate 28 residues downstream to its transmembrane domain (TMD) across protein-free bilayers (Brambillasca, S., M. Yabal, P. Soffientini, S. Stefanovic, M. Makarow, R.S. Hegde, and N. Borgese. 2005. EMBO J. 24:2533–2542). In the present study...

  17. Topological Domain Theory

    OpenAIRE

    Battenfeld, Ingo

    2008-01-01

    This thesis presents Topological Domain Theory as a powerful and flexible framework for denotational semantics. Topological Domain Theory models a wide range of type constructions and can interpret many computational features. Furthermore, it has close connections to established frameworks for denotational semantics, as well as to well-studied mathematical theories, such as topology and computable analysis.

  18. Modeling Protein Domain Function

    Science.gov (United States)

    Baker, William P.; Jones, Carleton "Buck"; Hull, Elizabeth

    2007-01-01

    This simple but effective laboratory exercise helps students understand the concept of protein domain function. They use foam beads, Styrofoam craft balls, and pipe cleaners to explore how domains within protein active sites interact to form a functional protein. The activity allows students to gain content mastery and an understanding of the…

  19. Domain Walls on Singularities

    CERN Document Server

    Halyo, Edi

    2009-01-01

    We describe domain walls that live on $A_2$ and $A_3$ singularities. The walls are BPS if the singularity is resolved and non--BPS if it is deformed and fibered. We show that these domain walls may interpolate between vacua that support monopoles and/or vortices.

  20. Chitinase-resistant hydrophilic symbiotic factors secreted by Frankia activate both Ca(2+) spiking and NIN gene expression in the actinorhizal plant Casuarina glauca.

    Science.gov (United States)

    Chabaud, Mireille; Gherbi, Hassen; Pirolles, Elodie; Vaissayre, Virginie; Fournier, Joëlle; Moukouanga, Daniel; Franche, Claudine; Bogusz, Didier; Tisa, Louis S; Barker, David G; Svistoonoff, Sergio

    2016-01-01

    Although it is now well-established that decorated lipo-chitooligosaccharide Nod factors are the key rhizobial signals which initiate infection/nodulation in host legume species, the identity of the equivalent microbial signaling molecules in the Frankia/actinorhizal association remains elusive. With the objective of identifying Frankia symbiotic factors we present a novel approach based on both molecular and cellular pre-infection reporters expressed in the model actinorhizal species Casuarina glauca. By introducing the nuclear-localized cameleon Nup-YC2.1 into Casuarina glauca we show that cell-free culture supernatants of the compatible Frankia CcI3 strain are able to elicit sustained high frequency Ca(2+) spiking in host root hairs. Furthermore, an excellent correlation exists between the triggering of nuclear Ca(2+) spiking and the transcriptional activation of the ProCgNIN:GFP reporter as a function of the Frankia strain tested. These two pre-infection symbiotic responses have been used in combination to show that the signal molecules present in the Frankia CcI3 supernatant are hydrophilic, of low molecular weight and resistant to chitinase degradation. In conclusion, the biologically active symbiotic signals secreted by Frankia appear to be chemically distinct from the currently known chitin-based rhizobial/arbuscular mycorrhizal signaling molecules. Convenient bioassays in Casuarina glauca are now available for their full characterization. PMID:26484850

  1. Intein-mediated purification of cytotoxic endonuclease I-TevI by insertional inactivation and pH-controllable splicing.

    Science.gov (United States)

    Wu, Wei; Wood, David W; Belfort, Georges; Derbyshire, Victoria; Belfort, Marlene

    2002-11-15

    An intein-mediated approach was developed for expression and affinity purification of a protein that is lethal to Escherichia coli. The protein, I-TevI, is an intron-encoded endonuclease. The approach involved the insertional inactivation of I-TevI with a controllable mini-intein placed in front of a cysteine required for splicing (an I-TevI::intein fusion). The purification was facilitated by a chitin-binding domain inserted into the mini-intein. Affinity purification of the I-TevI::intein fusion precursor on a chitin column was followed by pH-controllable splicing to restore the structure and function of I-TevI. To study the impact of the insertion context on I-TevI inactivation, the chimeric intein was inserted independently in front of seven cysteines of I-TevI. One of the seven intein integrants yielded I-TevI of high activity. This technique is, in principle, generalizable to the expression and purification of other cytotoxic proteins and is amenable to scale-up. PMID:12433989

  2. Urethral catheter insertion forces: a comparison of experience and training

    Directory of Open Access Journals (Sweden)

    Benjamin K. Canales

    2009-02-01

    Full Text Available Purpose: This study was undertaken to evaluate the insertion forces utilized during simulated placement of a urethral catheter by healthcare individuals with a variety of catheter experience. Materials and Methods: A 21F urethral catheter was mounted to a metal spring. Participants were asked to press the tubing spring against a force gauge and stop when they met a level of resistance that would typically make them terminate a catheter placement. Simulated catheter insertion was repeated fives times, and peak compression forces were recorded. Healthcare professionals were divided into six groups according to their title: urology staff, non-urology staff, urology resident/ fellow, non-urology resident/ fellow, medical student, and registered nurse. Results: A total of fifty-seven healthcare professionals participated in the study. Urology staff (n = 6 had the lowest average insertion force for any group at 6.8 ± 2.0 Newtons (N. Medical students (n = 10 had the least amount of experience (1 ± 0 years and the highest average insertion force range of 10.1 ± 3.7 N. Health care workers with greater than 25 years experience used significantly less force during catheter insertions (4.9 ± 1.8 N compared to all groups (p < 0.01. Conclusions: We propose the maximum force that should be utilized during urethral catheter insertion is 5 Newtons. This force deserves validation in a larger population and should be considered when designing urethral catheters or creating catheter simulators. Understanding urethral catheter insertion forces may also aid in establishing competency parameters for health care professionals in training.

  3. Feature-Level Domain Adaptation

    OpenAIRE

    Kouw, Wouter M.; Krijthe, Jesse H.; Loog, Marco; van der Maaten, Laurens J. P.

    2015-01-01

    Domain adaptation is the supervised learning setting in which the training and test data originate from different domains: the so-called source and target domains. In this paper, we propose and study a domain adaption approach, called feature-level domain adaptation (flda), that models the dependence between two domains by means of a feature-level transfer distribution. The domain adapted classifier is trained by minimizing the expected loss under this transfer distribution. Our empirical eva...

  4. The SAH domain extends the functional length of the myosin lever

    OpenAIRE

    Baboolal, TG; Sakamoto, T.; Forgacs, E; White, HD; Jackson, SM; Takagi, Y.; Farrow, RE; Molloy, JE; Knight, PJ; Sellers, JS; Peckham, M.

    2009-01-01

    Stable, single alpha-helix (SAH) domains are widely distributed in the proteome, including in myosins, but their functions are unknown. To test whether SAH domains can act as levers, we replaced four of the six calmodulin-binding IQ motifs in the levers of mouse myosin 5a (Myo5) with the putative SAH domain of Dictyostelium myosin MyoM of similar length. The SAH domain was inserted between the IQ motifs and the coiled coil in a Myo5 HMM construct in which the levers were truncated from six to...

  5. Some physical and chemical indices of clique-inserted lattices

    Science.gov (United States)

    Zhang, Zuhe

    2013-10-01

    The operation of replacing every vertex of an r-regular lattice H by a complete graph of order r is called clique-insertion, and the resulting lattice is called the clique-inserted lattice of H. For any given r-regular lattice, applying this operation iteratively, an infinite family of r-regular lattices is generated. Some interesting lattices including the 3-12-12 lattice can be constructed this way. In this paper, we recall the relationship between the spectra of an r-regular lattice and that of its clique-inserted lattice, and investigate the graph energy and resistance distance statistics. As an application, the asymptotic energy per vertex and average resistance distance of the 3-12-12 and 3-6-24 lattices are computed. We also give formulae expressing the numbers of spanning trees and dimer coverings of the kth iterated clique-inserted lattices in terms of those of the original one. Moreover, we show that new families of expander graphs can be constructed from the known ones by clique-insertion.

  6. Characterization and performance of alumina inserts reinforced with zirconia

    International Nuclear Information System (INIS)

    Alumina-based ceramics reinforced with zirconia were produced as inserts to be used as cutting tools. The objective of this work was to characterize and to evaluate the inserts as well as their performance in service. Additions of zirconia varying from 0.05 up to 0.20 wt% were done in alumina in order to select that composition presenting the best mechanical behavior. To do this, four-point bending mechanical strength, hardness and fracture toughness measurements were carried out. The best performance was found for alumina processing 0.010 wt.% of zirconia addition. The addition of zirconia was made with the objective of giving higher toughness to the insert throughout the microcracking reinforcing mechanism. The alumina reinforced inserts were compared with inserts of alumina without zirconia. It was accomplished by performing machining tests onto a SAE 1045 steel. The obtained results indicated that the zirconia reinforcement was able to improve the performance of the alumina cutting tools considerably. (author)

  7. Clinical outcome of narrow diameter implants inserted into allografts

    Directory of Open Access Journals (Sweden)

    Maurizio Franco

    2009-08-01

    Full Text Available OBJECTIVE: Narrow diameter implants (NDI (i.e. diameter <3.75 mm are a potential solution for specific clinical situations, such as reduced interradicular bone, thin alveolar crest and replacement of teeth with small cervical diameter. NDI have been available in clinical practice since the 1990s, but only few studies have analyzed their clinical outcome and no study have investigated NDI inserted in fresh-frozen bone (FFB grafts. Thus, a retrospective study on a series of NDI placed in homologue FFB was designed to evaluate their clinical outcome. MATERIAL AND METHODS: In the period between December 2003 and December 2006, 36 patients (22 females and 14 males, mean age 53 years with FFB grafts were selected and 94 different NDI were inserted. The mean follow-up was 25 months. To evaluate the effect of several host-, surgery-, and implant-related factors, marginal bone loss (MBL was considered an indicator of success rate (SCR. The Kaplan Meier algorithm and Cox regression were used. RESULTS: Only 5 out of 94 implants were lost (i.e. survival rate - SVR 95.7% and no differences were detected among the studied variables. On the contrary, the Cox regression showed that the graft site (i.e. maxilla reduced MBL. CONCLUSIONS: NDI inserted in FFB have a high SVR and SCR similar to those reported in previous studies on regular and NDI inserted in non-grafted jaws. Homologue FFB is a valuable material in the insertion of NDI.

  8. Insertion of nanoparticles into polymer brush under variable solvent conditions

    Science.gov (United States)

    Egorov, S. A.

    2012-10-01

    In this work, two-dimensional lattice-based self-consistent field theory is used to study the free energy cost associated with the insertion of a nanoparticle into a polymer brush. The nanoparticle is modeled as a cylinder and the self-consistent field equations are formulated on a cylindrical lattice. The use of two-dimensional formalism makes it possible to take into account the distortion of the brush density profile due to the embedded nanoinclusion. The insertion free energy penalty is analyzed as a function of the particle size, the brush grafting density, and the solvent quality. In agreement with the earlier simulation work, we find that the insertion free energy cost increases both with the particle size and the brush grafting density and decreases with deteriorating solvent quality. For nanoparticles located deep inside the brush, the insertion free energy is shown to scale with either squared (good solvent) or cubed (poor solvent) monomer volume fraction profile, in agreement with the earlier theoretical results. For shallow nanoinclusions located close to the brush surface, the insertion free energy is shown to scale linearly with the monomer volume fraction profile under both good and theta solvent conditions, again in agreement with the earlier work.

  9. Customizable engineered blood vessels using 3D printed inserts.

    Science.gov (United States)

    Pinnock, Cameron B; Meier, Elizabeth M; Joshi, Neeraj N; Wu, Bin; Lam, Mai T

    2016-04-15

    Current techniques for tissue engineering blood vessels are not customizable for vascular size variation and vessel wall thickness. These critical parameters vary widely between the different arteries in the human body, and the ability to engineer vessels of varying sizes could increase capabilities for disease modeling and treatment options. We present an innovative method for producing customizable, tissue engineered, self-organizing vascular constructs by replicating a major structural component of blood vessels - the smooth muscle layer, or tunica media. We utilize a unique system combining 3D printed plate inserts to control construct size and shape, and cell sheets supported by a temporary fibrin hydrogel to encourage cellular self-organization into a tubular form resembling a natural artery. To form the vascular construct, 3D printed inserts are adhered to tissue culture plates, fibrin hydrogel is deposited around the inserts, and human aortic smooth muscle cells are then seeded atop the fibrin hydrogel. The gel, aided by the innate contractile properties of the smooth muscle cells, aggregates towards the center post insert, creating a tissue ring of smooth muscle cells. These rings are then stacked into the final tubular construct. Our methodology is robust, easily repeatable and allows for customization of cellular composition, vessel wall thickness, and length of the vessel construct merely by varying the size of the 3D printed inserts. This platform has potential for facilitating more accurate modeling of vascular pathology, serving as a drug discovery tool, or for vessel repair in disease treatment. PMID:26732049

  10. Overview of insertion device controls at the Advanced Photon Source

    Science.gov (United States)

    Ramanathan, Mohan; Smith, Martin; Grimmer, John; Merritt, Michael

    2002-03-01

    The Advanced Photon Source (APS) is a third-generation synchrotron with major emphasis on insertion device (ID) sources. Currently, there are 25 sectors instrumented out of a possible 35 ID sources. Most of the insertion devices are undulators. Beamlines have been using the ID radiation at the APS for more than five years. The control system of choice at the APS is the experimental physics and industrial control system (EPICS) (http://www.aps.anl.gov/epics). Based on operational experience, the ID control system has been completely revamped. During user operations, the beamline user has complete control of the insertion device. Various interfaces, from RS-232 to EPICS channel access, have been provided for the users to control the IDs. The control system software has been designed to accommodate scanning of the insertion device synchronized to each user's beamline monochromator. The users have the option of operating the device in a tapered mode. The control software allows the users to control the undulators in energy space from the fundamental to the seventh harmonic. The design philosophy of the insertion device control system will be discussed. The implementation and operational experience will be presented in detail.

  11. Some physical and chemical indices of clique-inserted lattices

    International Nuclear Information System (INIS)

    The operation of replacing every vertex of an r-regular lattice H by a complete graph of order r is called clique-insertion, and the resulting lattice is called the clique-inserted lattice of H. For any given r-regular lattice, applying this operation iteratively, an infinite family of r-regular lattices is generated. Some interesting lattices including the 3-12-12 lattice can be constructed this way. In this paper, we recall the relationship between the spectra of an r-regular lattice and that of its clique-inserted lattice, and investigate the graph energy and resistance distance statistics. As an application, the asymptotic energy per vertex and average resistance distance of the 3-12-12 and 3-6-24 lattices are computed. We also give formulae expressing the numbers of spanning trees and dimer coverings of the kth iterated clique-inserted lattices in terms of those of the original one. Moreover, we show that new families of expander graphs can be constructed from the known ones by clique-insertion. (paper)

  12. Recombination of an intrachromosomal paracentric insertion of chromosome 3

    Energy Technology Data Exchange (ETDEWEB)

    Best, R.G.; Burnett, W.J.; Brock, J.K. [Univ. of South Carolina School of Medicine, Columbia, SC (United States)] [and others

    1994-09-01

    Cytogenetic studies were initiated on a newborn female due to multiple congenital anomalies including microcephaly, clinodactyly, abnormal positioning of hands, left facial palsy, heart defect, sacral dimple, and facial dysmorphic features. Facial features were described as low set rotated ears, nystagmus, and a small, flattened nose. A structural rearrangement of the long arm of chromosome 3 was observed with a complex banding pattern. Study of parental chromosomes revealed a normal male pattern for the father, and an intrachromosomal insertion on the long arm of chromosome 3 for the mother described as 46,XX,dir ins(3)(q21q23q26.2). Further characterization of the proband`s structurally abnormal chromosome 3 revealed a karyotype best described as: 46,XX,rec(3),dupq23{r_arrow}q26.2::q21{r_arrow}q23,dir ins(3)(q21q23q26.2), which is a partial duplication of both the inserted segment as well as the intervening segment between the inserted segment and the insertion site. This would appear to be the result of a three-strand double cross-over within the insertion loop. Molecular cytogenetic studies are presently underway to further elucidate chromosome structure of the proband and her mother.

  13. Domain-Specific Multimodeling

    DEFF Research Database (Denmark)

    Hessellund, Anders

    Enterprise systems are complex artifacts. They are hard to build, manage, understand, and evolve. Existing software development paradigms fail to properly address challenges such as system size, domain complexity, and software evolution when development is scaled to enterprise systems. We propose...... domain-specific multimodeling as a development paradigm to tackle these challenges in a language-oriented manner. The different concerns of a system are conceptually separated and made explicit as independent domain-specific languages. This approach increases productivity and quality by raising the...

  14. Silver clusters insert into polymer solar cell for enhancing light absorption

    Institute of Scientific and Technical Information of China (English)

    Guolong Li; Hongyu Zhen; Zhuoyin Huang; Kan Li; Weidong Shen; Xu Liu

    2012-01-01

    As an employment of surface plasmonic effect,the consequence of insertion of a layer of Ag clusters into polymer solar cell on the enhancement of light absorption and power conversion efficiency is investigated.Optical analysis based on the finite-difference time-domain (FDTD) is performed with experiments to evaluate the effect of the interaction between the Ag clusters and incident light on light absorption in polymer solar cell Ag clusters modify the light wave vector and the electromagnetic field inside the device is redistributed and enhanced.As a result,polymer solar cells achieve an overall increase in absorption,short-circuit current density,and power conversion efficiency.

  15. Genetic Analysis of MEFV Gene Pyrin Domain in Patients With Behçet's Disease

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available Objectives. Behçet's disease (BD is a systemic vasculitis with recurrent oral and genital ulcers and uveitis. MEFV gene, which is the main factor in familial Mediterranean fever (FMF, is also reported to be a susceptibility gene for BD. The pyrin domain of MEFV gene is a member of death-domain superfamily and has been proposed to regulate inflammatory signaling in myeloid cells. This study was designed to determine if mutations in pyrin domain of MEFV gene are involved in BD. Methods. We analyzed the pyrin domain of MEFV gene in 54 Turkish patients with BD by PCR-analysis and direct sequencing. Results. Neither deletion or insertion mutations nor point mutations in pyrin domain were found in any patient. Conclusion. Although pyrin gene mutations have been reported in patients with BD, pyrin domain is not mutated. However, alterations in other regions of MEFV gene and interaction between pyrin domains are needed to be further investigated.

  16. Quantum chemical study on insertion and abstraction reaction of dichlorocarbene with methyl alcohol and methyl mercaptan

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The insertion and abstraction reaction mechanisms of singlet and triplet CCl2 with CH3MH (M=O, S) have been studied by using the DFT, NBO and AIM methods. The geometries of reactions, the transition state and products were completely optimized by B3LYP/6-311G(d, p). All the energy of the species was obtained at the CCSD(T)/6-311G(d, p) level. The calculated results indicated that the major pathways of the reaction were obtained on the singlet potential energy surface. The singlet CCl2 can not only trigger the insertion reaction with C-H and M-H in four pathways, by which the products P1 [CH3OCHCl2, reactionⅠ(1)], P3[Cl2HCCH2OH, reactionⅠ(2)], P5[CH3SCHCl2, reactionⅡ(1)] and P7[Cl2HCCH2SH, reac-tion Ⅱ(2)] are produced respectively, but also abstract M-H, resulting P4 [CH2O+CH2Cl2, reactionⅠ(3)] and P8[CH2S+CH2Cl2, reactionⅡ(3)]. In addition, the important geometries in domain pathways have been studied by AIM and NBO theories.

  17. Quantum chemical study on insertion and abstraction reaction of dichlorocarbene with methyl alcohol and methyl mercaptan

    Institute of Scientific and Technical Information of China (English)

    LI Zhi-Feng; ZHU Yuan-Cheng; LI Hui-Xue; LIU Xin-Wen; LU Xiao-Quan

    2009-01-01

    The Insertion and abstraction reaction mechanisms of singlet and triplet CCl2 with CH3MH (M=O, S) have been studied by using the DFT, NBO and AIM methods. The geometries of reactions, the transition state and products were completely optimized by B3LYP/6-311G(d, p). All the energy of the species was obtained at the CCSD(T)/6-311G(d, p) level. The calculated results indicated that the major pathways of the reaction were obtained on the singlet potential energy surface. The singlet CCl2 can not only trigger the insertion reaction with C-H and M-H in four pathways, by which the products P1 [CH3OCHCl2, reaction I (1)], P3[Cl2HCCH2OH, reaction Ⅰ(2)], PS[CH3SCHCl2, reaction Ⅱ(1)] and P7[Cl2HCCH2SH, reac-tion Ⅱ(2)] are produced respectively, but also abstract M-H, resulting P4 [CH2O+CH2Cl2, reaction Ⅰ(3)] and PS[CH2S+CH2Cl2, reaction Ⅱ (3)]. In addition, the important geometries in domain pathways have been studied by AIM and NBO theories.

  18. How to insert a nasogastric tube and check gastric position at the bedside.

    Science.gov (United States)

    Best, Carolyn

    2016-05-18

    Rationale and key points A nasogastric tube (NG tube) can be used to aspirate stomach contents or to administer feed, medication or fluid into the stomach. ▶ A blind technique is used to insert the NG tube through the nostril, along the nasopharynx, through the oesophagus and into the stomach. ▶ It is important for nurses to be able to recognise problems that may arise when inserting a NG tube blindly, and to know what actions to take if it is suspected that the distal tip of the NG tube is not sitting in the stomach, or they are unable to identify its location. ▶ Misplacement and subsequent use of a NG tube to administer feed, medication or fluid is a 'never event' ( NHS England Patient Safety Domain 2015 ). Reflective activity Clinical skills articles can help update your practice and ensure it remains evidence based. Apply this article to your practice. Reflect on and write a short account of: 1. How you think this article will change your practice. 2. How you could use this resource to educate your colleagues. Subscribers can update their reflective accounts at rcni.com/portfolio. PMID:27191450

  19. The strategic use of inserts in the Brazilian presidential elections

    Directory of Open Access Journals (Sweden)

    Felipe Borba

    2012-01-01

    Full Text Available The aim of this article is to analyze the communication strategies of presidential candidates during the elections held in 2006 and 2010. The focus is on the strategic component of electoral inserts and the methodology consists of investigating how candidates choose to distribute these inserts in the programming of television networks. The results indicate that the candidates pursue different strategies influenced basically by three variables: electoral legislation, their standing in polls and the difference of resources available. In parallel, the article debates the role of the regulation of electoral advertising and how this set of rules influences the level of information of campaigns, the occurrence of attacks, and party strategies. Overall, 2,993 electoral inserts were examined.

  20. Assessment of plutonium and minor actinides insertions in HTR

    International Nuclear Information System (INIS)

    Based in the specifications of the high temperature reactor - HTR developed by H. J. Ruetten and K. Haas (Nucl. Eng. and Design: 195, 353-360, 2000), it was studied the possibility of insertion of minor actinides in this type of reactor. In this first study, carried out with the WIMSD5 code, the effective multiplication factor and the temperature reactivity coefficient had been evaluated. The behavior of the multiplication factor with fuel burnup for the standard fuel composition (with PuO2) as well as for the case with the insertion of minor actinides originated from a PWR spent fuel, was as expected. The results suggest the possibility of insertion of joint plutonium and minor actinides in the fuel composition. (author)

  1. Visualizing Knowledge Domains.

    Science.gov (United States)

    Borner, Katy; Chen, Chaomei; Boyack, Kevin W.

    2003-01-01

    Reviews visualization techniques for scientific disciplines and information retrieval and classification. Highlights include historical background of scientometrics, bibliometrics, and citation analysis; map generation; process flow of visualizing knowledge domains; measures and similarity calculations; vector space model; factor analysis;…

  2. Conserved Domain Database (CDD)

    Data.gov (United States)

    U.S. Department of Health & Human Services — CDD is a protein annotation resource that consists of a collection of well-annotated multiple sequence alignment models for ancient domains and full-length proteins.

  3. Metallocene Catalytic Insertion Polymerization of 1-Silene to Polycarbosilanes

    Science.gov (United States)

    Tian, Yuelong; Ge, Min; Zhang, Weigang; Lv, Xiaoxu; Yu, Shouquan

    2015-11-01

    Metallocene of zirconium were used as a catalyst for an insertion polymerization of 1-methylsilene directly into pre-ceramic precursor polyzirconocenecarbosilane (PZCS) during dechlorination of dichlorodimethylesilane by sodium, which exhibits high catalytic effectiveness with the maximum conversion ratio of polycarbosilane up to 91%. The average molecular weights of polymers synthesized are less than 1400, all with very narrow polymolecularities. The mechanism of catalytic polymerization was assumed to be similar to a coordination insertion polymerization of 1-olefins by metallocenes. The obtained PZCS show high ceramic yields with formation of composite ceramics of ZrC-SiC, which are novel polymeric precursors of ultra-high temperature ceramic (UHTC) fiber and composite.

  4. Magnetic ripple correction in tandem mirrors by ferromagnetic inserts

    International Nuclear Information System (INIS)

    Magnetic ripple of 1% or more caused by discrete solenoid coils in the central cells of tandem mirrors may severely affect the MHD stability. The ripple amplitude can be reduced by an order of magnitude by ferromagnetic annuli inserted within the coils at the regions of ripple maxima. The inserts need not affect the accessibility, coil diameter, or capital cost, since large quantities of steel are required within the coils for the neutron blanket and shield. Design of the ripple correction is simplified and linearized by the cylindrical geometry and by the saturation of the ferromagnetic steel

  5. Tension Pneumothorax and Subcutaneous Emphysema Complicating Insertion of Nasogastric Tube

    Directory of Open Access Journals (Sweden)

    Narjis AL Saif

    2015-01-01

    Full Text Available Nasogastric tube has a key role in the management of substantial number of hospitalized patients particularly the critically ill. In spite of the apparent simple insertion technique, nasogastric tube placement has its serious perhaps fatal complications which need to be carefully assessed. Pulmonary misplacement and associated complications are commonplace during nasogastric tube procedure. We present a case of tension pneumothorax and massive surgical emphysema in critically ill ventilated patient due to inadvertent nasogastric tube insertion and also discussed the risk factors, complication list, and arrays of techniques for safer tube placement.

  6. Rectal fist insertion. An unusual form of sexual behavior.

    Science.gov (United States)

    Shook, L L; Whittle, R; Rose, E F

    1985-12-01

    Rectal fist insertion (fist fucking) is an uncommon and potentially dangerous sexual practice. This is usually a homosexual activity, but can also be a heterosexual or an autoerotic practice. One known death has been reported associated with rectal fist insertion, in which the complications of anal and colonic tears and bleeding had occurred (see Editor's note). The possibility of drug overdose is also probable, as drugs and alcohol are commonly introduced into the rectum to promote sphincter relaxation and to ease the discomfort of anal dilatation. PMID:4072987

  7. Tension Pneumothorax and Subcutaneous Emphysema Complicating Insertion of Nasogastric Tube

    Science.gov (United States)

    AL Saif, Narjis; Hammodi, Adel; Al-Azem, M. Ali; Al-Hubail, Rasheed

    2015-01-01

    Nasogastric tube has a key role in the management of substantial number of hospitalized patients particularly the critically ill. In spite of the apparent simple insertion technique, nasogastric tube placement has its serious perhaps fatal complications which need to be carefully assessed. Pulmonary misplacement and associated complications are commonplace during nasogastric tube procedure. We present a case of tension pneumothorax and massive surgical emphysema in critically ill ventilated patient due to inadvertent nasogastric tube insertion and also discussed the risk factors, complication list, and arrays of techniques for safer tube placement. PMID:26448883

  8. Improving the chromatic effects of storage rings with antisymmetric insertions

    International Nuclear Information System (INIS)

    High luminosity storage rings require good chromatic behavior for beams with large momentum spreads. This requires that the effects of half-integer structure resonances for off-momentum particles be minimized. We show that a lattice with antisymmetric insertions can be so designed that the driving term for the half-integer structure resonance is suppressed by cancellation of successive pairs of high-beta multiplets. Hence, even though the periodicity is half that of a lattice with symmetric insertions, the chromatic properties are similar

  9. Insertion of lithium into electrochromic devices after completion

    Science.gov (United States)

    Berland, Brian Spencer; Lanning, Bruce Roy; Frey, Jonathan Mack; Barrett, Kathryn Suzanne; DuPont, Paul Damon; Schaller, Ronald William

    2015-12-22

    The present disclosure describes methods of inserting lithium into an electrochromic device after completion. In the disclosed methods, an ideal amount of lithium can be added post-fabrication to maximize or tailor the free lithium ion density of a layer or the coloration range of a device. Embodiments are directed towards a method to insert lithium into the main device layers of an electrochromic device as a post-processing step after the device has been manufactured. In an embodiment, the methods described are designed to maximize the coloration range while compensating for blind charge loss.

  10. Ultrasonic airborne insertion loss measurements at normal incidence (L).

    Science.gov (United States)

    Farley, Jayrin; Anderson, Brian E

    2010-12-01

    Transmission loss and insertion loss measurements of building materials at audible frequencies are commonly made using plane wave tubes or as a panel between reverberant rooms. These measurements provide information for noise isolation control in architectural acoustics and in product development. Airborne ultrasonic sound transmission through common building materials has not been fully explored. Technologies and products that utilize ultrasonic frequencies are becoming increasingly more common, hence the need to conduct such measurements. This letter presents preliminary measurements of the ultrasonic insertion loss levels for common building materials over a frequency range of 28-90 kHz using continuous-wave excitation. PMID:21218864

  11. Shield Insertion to Minimize Noise Amplitude in Global Interconnects

    Directory of Open Access Journals (Sweden)

    Kalpana.A.B

    2012-09-01

    Full Text Available Shield insertion is an effective technique for minimise crosstalk noise and signal delay uncertainty .To reduce the effects of coupling uniform or simultaneous shielding may be used on either or both sides of a signal line. Shields are ground or power lines placed between two signal wires to prevent direct coupling between them as the shield width increases, the noise amplitude decreases, in this paper inserting a shield line between two coupled interconnects is shown to be more effective in reducing crosstalk noise for different technology nodes .

  12. Advanced photon source experience with vacuum chambers for insertion devices

    International Nuclear Information System (INIS)

    During the last five years, a new approach to the design and fabrication of extruded aluminum vacuum chambers for insertion devices was developed at the Advanced Photon Source (APS). With this approach, three different versions of the vacuum chamber, with vertical apertures of 12 mm, 8 mm, and 5 mm, were manufactured and tested. Twenty chambers were installed into the APS vacuum system. All have operated with beam, and 16 have been coupled with insertion devices. Two different vacuum chambers with vertical apertures of 16 mm and 11 mm were developed for the BESSY-II storage ring and 3 of 16 mm chambers were manufactured

  13. The supermodule insertion tool of the CMS electromagnetic calorimeter and the first trial insertion of a supermodule.

    CERN Multimedia

    Maximilien Brice

    2006-01-01

    The first trial insertion of a complete Electromagnetic Calorimeter (ECAL) "supermodule" (1700 lead-tungstate crystals, with support structures, light detectors (avalanche photodiodes), readout electronics and cooling system) was performed on 1st March. This delicate operation - sliding a 2-tonne 3m-long object onto support rails (in real life these are attached to the barrel hadron calorimeter (HCAL)) - made use of a custom designed "squirrel cage". The rotatable squirrel cage allows the insertion of any supermodule into any of the 18 positions, including very fine (sub-mm) adjustments. The first supermodule will be inserted into the real HCAL later this month in preparation for the "magnet test and cosmic-ray challenge" (MTCC). In the first image the supermodule is in the centre and the alignment disks are highlighted by the flash.

  14. Chitinase 3-Like 1 (Chil1) Regulates Survival and Macrophage-Mediated Interleukin-1β and Tumor Necrosis Factor Alpha during Pseudomonas aeruginosa Pneumonia.

    Science.gov (United States)

    Marion, Chad R; Wang, Jianmiao; Sharma, Lokesh; Losier, Ashley; Lui, Wei; Andrews, Nathaniel; Elias, Jack A; Kazmierczak, Barbara I; Roy, Craig R; Dela Cruz, Charles S

    2016-07-01

    Pseudomonas aeruginosa causes hospital-acquired pneumonia and is associated with high mortality. An effective response to such an infection includes efficient clearance of pathogenic organisms while limiting collateral damage from the host inflammatory response, known as host resistance and host tolerance, respectively. P. aeruginosa expresses a type III secretion system (T3SS) needle complex that induces NLRC4 (NOD-like receptor C4) activation, interleukin-1β (IL-1β) production, and host tissue damage. Chitinase 3-like-1 (Chil1) is expressed during infection and binds to its receptor, IL-13 receptor α2 (IL-13Rα2), to regulate the pathogen-host response during Streptococcus pneumoniae infection, but the role Chil1 plays in balancing the host resistance and host tolerance during P. aeruginosa pneumonia is not known. We conducted experiments using C57BL/6 mice with or without a genetic deficiency of Chil1 and demonstrated that Chil1-deficient mice succumb to P. aeruginosa infection more rapidly than the wild type (WT). The decreased survival time in infected Chil1-deficient mice is associated with more neutrophils recruited to the airways, more lung parenchymal damage, and increased pulmonary consolidation while maintaining equivalent bacterial killing compared to WT mice. Infected Chil1-deficient mice and bone marrow-derived macrophages (BMDMs) from Chil1-deficient mice have increased production of tumor necrosis factor alpha (TNF-α) and IL-1β compared to infected WT mice and macrophages. Infection of Chil1-deficient BMDMs with non-NLRC4-triggering P. aeruginosa, which is deficient in the T3SS needle complex, did not alter the excessive IL-1β production compared to BMDMs from WT mice. The addition of recombinant Chil1 decreases the excessive IL-1β production but only partially rescues stimulated BMDMs from IL-13Rα2-deficient mice. Our data provide mechanistic insights into how Chil1 regulates P. aeruginosa-induced host responses. PMID:27141083

  15. BjMYB1, a transcription factor implicated in plant defence through activating BjCHI1 chitinase expression by binding to a W-box-like element

    Science.gov (United States)

    Gao, Ying; Jia, Shuangwei; Wang, Chunlian; Wang, Fujun; Wang, Fajun; Zhao, Kaijun

    2016-01-01

    We previously identified the W-box-like-4 (Wbl-4) element (GTAGTGACTCAT), one of six Wbl elements in the BjC-P promoter of the unusual chitinase gene BjCHI1 from Brassica juncea, as the core element responsive to fungal infection. Here, we report the isolation and characterization of the cognate transcription factor interacting with the Wbl-4 element. Using Wbl-4 as a target, we performed yeast one-hybrid screening of a B. juncea cDNA library and isolated an R2R3-MYB transcription factor designated as BjMYB1. BjMYB1 was localized in the nucleus of plant cells. EMSA assays confirmed that BjMYB1 binds to the Wbl-4 element. Transiently expressed BjMYB1 up-regulated the activity of the BjC-P promoter through its binding to the Wbl-4 element in tobacco (Nicotiana benthamiana) leaves. In B. juncea, BjMYB1 displayed a similar induced expression pattern as that of BjCHI1 upon infection by the fungus Botrytis cinerea. Moreover, heterogeneous overexpression of BjMYB1 significantly elevated the resistance of transgenic Arabidopsis thaliana to the fungus B. cinerea. These results suggest that BjMYB1 is potentially involved in host defence against fungal attack through activating the expression of BjCHI1 by binding to the Wbl-4 element in the BjC-P promoter. This finding demonstrates a novel DNA target of plant MYB transcription factors. PMID:27353280

  16. Glucanases and Chitinases as Causal Agents in the Protection of Acacia Extrafloral Nectar from Infestation by Phytopathogens1[W][OA

    Science.gov (United States)

    González-Teuber, Marcia; Pozo, María J.; Muck, Alexander; Svatos, Ales; Adame-Álvarez, Rosa M.; Heil, Martin

    2010-01-01

    Nectars are rich in primary metabolites and attract mutualistic animals, which serve as pollinators or as an indirect defense against herbivores. Their chemical composition makes nectars prone to microbial infestation. As protective strategy, floral nectar of ornamental tobacco (Nicotiana langsdorffii × Nicotiana sanderae) contains “nectarins,” proteins producing reactive oxygen species such as hydrogen peroxide. By contrast, pathogenesis-related (PR) proteins were detected in Acacia extrafloral nectar (EFN), which is secreted in the context of defensive ant-plant mutualisms. We investigated whether these PR proteins protect EFN from phytopathogens. Five sympatric species (Acacia cornigera, A. hindsii, A. collinsii, A. farnesiana, and Prosopis juliflora) were compared that differ in their ant-plant mutualism. EFN of myrmecophytes, which are obligate ant-plants that secrete EFN constitutively to nourish specialized ant inhabitants, significantly inhibited the growth of four out of six tested phytopathogenic microorganisms. By contrast, EFN of nonmyrmecophytes, which is secreted only transiently in response to herbivory, did not exhibit a detectable inhibitory activity. Combining two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with nanoflow liquid chromatography-tandem mass spectrometry analysis confirmed that PR proteins represented over 90% of all proteins in myrmecophyte EFN. The inhibition of microbial growth was exerted by the protein fraction, but not the small metabolites of this EFN, and disappeared when nectar was heated. In-gel assays demonstrated the activity of acidic and basic chitinases in all EFNs, whereas glucanases were detected only in EFN of myrmecophytes. Our results demonstrate that PR proteins causally underlie the protection of Acacia EFN from microorganisms and that acidic and basic glucanases likely represent the most important prerequisite in this defensive function. PMID:20023149

  17. How does the Shift-insertion sort behave when the sorting elements follow a Normal distribution?

    CERN Document Server

    Pal, Mita; Mahanti, N C

    2012-01-01

    The present paper examines the behavior of Shift-insertion sort (insertion sort with shifting) for normal distribution inputs and is in continuation of our earlier work on this new algorithm for discrete distribution inputs, namely, negative binomial. Shift insertion sort is found more sensitive for main effects but not for all interaction effects compared to conventional insertion sort.

  18. Quench Module Insert Capabilities and Development Test Results

    Science.gov (United States)

    Carswell, B.; Crouch, M.; Farmer, J.; Breeding, S.; Rose, F.; Whitaker, Ann F. (Technical Monitor)

    2001-01-01

    Quench Module Insert is a directional solidification furnace, which will fly in the Materials Science Research Facility. The QMI provides high thermal gradient and quench capabilities for processing metals and alloys in microgravity. This paper will describe the capabilities and present of on-going analysis and development testing.

  19. Otological Findings Ten Years after Myringotomy with Tympanostomy Tube Insertion

    Directory of Open Access Journals (Sweden)

    Ali Goljanian Tabrizi

    2011-01-01

    Full Text Available Introduction: To study the long-term complications of tympanostomy tube insertion in young children 10 years after surgery.   Materials and Methods: In September 2011, the medical records of all patients who had undergone myringotomy with tympanostomy tube insertion between February 2000 and March 2001 at the two general hospitals of Isfahan University of Medical Sciences were studied. Of the 98 patients who fulfilled the inclusion criteria, 82 patients agreed to participate and were enrolled in the study. The complications of the operation were evaluated in these patients.   Results: Of the 164 ears that were operated on, myringosclerosis was found in 17.1%, atrophy of the tympanic membrane in 1.2%, permanent perforation of the tympanic membrane in 0.6% and tympanic membrane atelectasis in 0.6%. None of the patients developed cholesteatoma as a complication of tympanostomy tube insertion.   Conclusion:  Considering the low risk of serious complications after 10 years, tympanostomy tube insertion is a safe and effective treatment option in the treatment of otitis media with effusion.

  20. Prototype ISR Superconducting Quadrupole for the low beta insertion.

    CERN Multimedia

    1976-01-01

    The four coils are provisionally kept together by aluminium clamps while epoxy-glass bands are wrapped around them to form a number of spacer rings.Stainless steel spacers were then inserted between these rings and the yoke quadrants. The persons are Michel Bouvier(left) and Pierre Pugin. See also7702690X.

  1. Discovery of unfixed endogenous retrovirus insertions in diverse human populations.

    Science.gov (United States)

    Wildschutte, Julia Halo; Williams, Zachary H; Montesion, Meagan; Subramanian, Ravi P; Kidd, Jeffrey M; Coffin, John M

    2016-04-19

    Endogenous retroviruses (ERVs) have contributed to more than 8% of the human genome. The majority of these elements lack function due to accumulated mutations or internal recombination resulting in a solitary (solo) LTR, although members of one group of human ERVs (HERVs), HERV-K, were recently active with members that remain nearly intact, a subset of which is present as insertionally polymorphic loci that include approximately full-length (2-LTR) and solo-LTR alleles in addition to the unoccupied site. Several 2-LTR insertions have intact reading frames in some or all genes that are expressed as functional proteins. These properties reflect the activity of HERV-K and suggest the existence of additional unique loci within humans. We sought to determine the extent to which other polymorphic insertions are present in humans, using sequenced genomes from the 1000 Genomes Project and a subset of the Human Genome Diversity Project panel. We report analysis of a total of 36 nonreference polymorphic HERV-K proviruses, including 19 newly reported loci, with insertion frequencies ranging from 0.75 that varied by population. Targeted screening of individual loci identified three new unfixed 2-LTR proviruses within our set, including an intact provirus present at Xq21.33 in some individuals, with the potential for retained infectivity. PMID:27001843

  2. Engine valve and seat insert wear study with a simulator

    Institute of Scientific and Technical Information of China (English)

    Y.S.Wang; S.Narasimhan

    2001-01-01

    The demands on higher performance and the increasing use of alternative fuels chal-lenge engine valves now with greater wear problems than before. A seat wear simulator was builtto evaluate the compatibility and wear of valve and seat insert. The rig test results have been suc-cessfully correlated with engine test results. In this study, intake valves made from Sil 1 materialwere treated with salt bath nitride processes and tested against six different insert materials. Wearresistance of these combinations was ranked and compared to the Sil 1 valve without nitriding.The test results demonstrate that nitriding improved valve seat wear resistance. In the total valveseat recession ranking, the combination of nitrided Sil 1 valve against T 400 insert exhibited theleast total recession among the nineteen combinations of valve and insert tested. The results indi-cate that the valve seat wear mechanisms are a complex combination of adhesion and shearstrain. The nitrides in the compound layer of nitrided valves gave strong atomic bonding, higherhardness, compressive residual stresses, and possible low friction, thus resulted in the superiorwear performance.

  3. Chemo-port insertion: A cause of vocal cord palsy.

    Science.gov (United States)

    Alazzawi, Sarmad; Hindi, Khalid; Malik, Ausama; Wee, Chong Aun; Prepageran, Narayanan

    2015-11-01

    We describe extremely rare cases of vocal cord palsy following surgical insertion of a chemo port. Our cohort consisted of patients with cancer who developed hoarseness immediately after central venous line placement for the administration of chemotherapy, with vocal cord palsy confirmed with flexible laryngoscopy. Given the timing, central venous line placement appears to be the most likely cause. PMID:26108861

  4. Insertion of lithium in Nb2O5

    International Nuclear Information System (INIS)

    The electrode process during the reduction and oxidation of Nb sub(2) O sub(5) by cyclic voltametry was characterized. The surface analysis by X-ray photo electronic spectroscopy allowed characterize the surface processes, that happening during the insertion reaction. (author)

  5. Effects of Rotational Motion in Robotic Needle Insertion

    Science.gov (United States)

    Ramezanpour, H.; Yousefi, H.; Rezaei, M.; Rostami, M.

    2015-01-01

    Background Robotic needle insertion in biological tissues has been known as one the most applicable procedures in sampling, robotic injection and different medical therapies and operations. Objective In this paper, we would like to investigate the effects of angular velocity in soft tissue insertion procedure by considering force-displacement diagram. Non-homogenous camel liver can be exploited as a tissue sample under standard compression test with Zwick/Roell device employing 1-D axial load-cell. Methods Effects of rotational motion were studied by running needle insertion experiments in 5, 50 and 200 mm/min in two types of with or without rotational velocity of 50, 150 and 300 rpm. On further steps with deeper penetrations, friction force of the insertion procedure in needle shaft was acquired by a definite thickness of the tissue. Results Designed mechanism of fixture for providing different frequencies of rotational motion is available in this work. Results for comparison of different force graphs were also provided. Conclusion Derived force-displacement graphs showed a significant difference between two procedures; however, tissue bleeding and disorganized micro-structure would be among unavoidable results. PMID:26688800

  6. Effects of lithium insertion on thermal conductivity of silicon nanowires

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Wen [Department of Physics, Centre for Advanced 2D Material and Centre for Computational Science and Engineering, National University of Singapore, Singapore, Singapore 117546 (Singapore); Institute of High Performance Computing, A*STAR, Singapore, Singapore 138632 (Singapore); Zhang, Gang, E-mail: zhangg@ihpc.a-star.edu.sg [Institute of High Performance Computing, A*STAR, Singapore, Singapore 138632 (Singapore); Li, Baowen [Department of Physics, Centre for Advanced 2D Material and Centre for Computational Science and Engineering, National University of Singapore, Singapore, Singapore 117546 (Singapore); NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Kent Ridge, Singapore 119620 (Singapore); Center for Phononics and Thermal Energy Science, School of Physics Science and Engineering, Tongji University, 200092 Shanghai (China)

    2015-04-27

    Recently, silicon nanowires (SiNWs) have been applied as high-performance Li battery anodes, since they can overcome the pulverization and mechanical fracture during lithiation. Although thermal stability is one of the most important parameters that determine safety of Li batteries, thermal conductivity of SiNWs with Li insertion remains unclear. In this letter, using molecular dynamics simulations, we study room temperature thermal conductivity of SiNWs with Li insertion. It is found that compared with the pristine SiNW, there is as much as 60% reduction in thermal conductivity with 10% concentration of inserted Li atoms, while under the same impurity concentration the reduction in thermal conductivity of the mass-disordered SiNW is only 30%. With lattice dynamics calculations and normal mode decomposition, it is revealed that the phonon lifetimes in SiNWs decrease greatly due to strong scattering of phonons by vibrational modes of Li atoms, especially for those high frequency phonons. The observed strong phonon scattering phenomenon in Li-inserted SiNWs is similar to the phonon rattling effect. Our study serves as an exploration of thermal properties of SiNWs as Li battery anodes or weakly coupled with impurity atoms.

  7. Magnesium oxide inserts for the LECO Carbon Analyzer

    Energy Technology Data Exchange (ETDEWEB)

    Bagaasen, L.M.; Jensen, C.M.

    1991-01-16

    LECO carbon analysis of plutonium metal and plutonium oxide at the Rocky Flats Plant generates several hundred kilograms of high silica residues each year. The plutonium in these residues is difficult and expensive to recover using production dissolution processes. A magnesium oxide (MgO) insert has been developed that significantly lowers the plutonium recovery costs without adversely affecting accuracy of the carbon analysis.

  8. Effects of lithium insertion on thermal conductivity of silicon nanowires

    International Nuclear Information System (INIS)

    Recently, silicon nanowires (SiNWs) have been applied as high-performance Li battery anodes, since they can overcome the pulverization and mechanical fracture during lithiation. Although thermal stability is one of the most important parameters that determine safety of Li batteries, thermal conductivity of SiNWs with Li insertion remains unclear. In this letter, using molecular dynamics simulations, we study room temperature thermal conductivity of SiNWs with Li insertion. It is found that compared with the pristine SiNW, there is as much as 60% reduction in thermal conductivity with 10% concentration of inserted Li atoms, while under the same impurity concentration the reduction in thermal conductivity of the mass-disordered SiNW is only 30%. With lattice dynamics calculations and normal mode decomposition, it is revealed that the phonon lifetimes in SiNWs decrease greatly due to strong scattering of phonons by vibrational modes of Li atoms, especially for those high frequency phonons. The observed strong phonon scattering phenomenon in Li-inserted SiNWs is similar to the phonon rattling effect. Our study serves as an exploration of thermal properties of SiNWs as Li battery anodes or weakly coupled with impurity atoms

  9. Modeling of Porous Insertion Electrodes with Liquid Electrolyte

    DEFF Research Database (Denmark)

    West, Keld; Jacobsen, Torben; Atlung, Sven

    1982-01-01

    The dynamics of porous insertion electrodes during charge or discharge is described by a simplified mathematicalmodel, accounting for the coupled transport in electrode and electrolyte phases. A numerical method to evaluate theresponse of this model to either controlled potential or controlled cu...

  10. Root cause of incomplete control rod insertions at Westinghouse reactors

    Energy Technology Data Exchange (ETDEWEB)

    Ray, S. [Westinghouse, Monroeville, PA (United States)

    1997-01-01

    Within the past year, incomplete RCCA insertions have been observed on high burnup fuel assemblies at two Westinghouse PWRs. Initial tests at the Wolf Creek site indicated that the direct cause of the incomplete insertions observed at Wolf Creek was excessive fuel assembly thimble tube distortion. Westinghouse committed to the NRC to perform a root cause analysis by the end of August, 1996. The root cause analysis process used by Westinghouse included testing at ten sites to obtain drag, growth and other characteristics of high burnup fuel assemblies. It also included testing at the Westinghouse hot cell of two of the Wolf Creek incomplete insertion assemblies. A mechanical model was developed to calculate the response of fuel assemblies when subjected to compressive loads. Detailed manufacturing reviews were conducted to determine if this was a manufacturing related issue. In addition, a review of available worldwide experience was performed. Based on the above, it was concluded that the thimble tube distortion observed on the Wolf Creek incomplete insertion assemblies was caused by unusual fuel assembly growth over and above what would typically be expected as a result of irradiation exposure. It was determined that the unusual growth component is a combination of growth due to oxide accumulation and accelerated growth, and would only be expected in high temperature plants on fuel assemblies that see long residence times and high power duties.

  11. Folding and insertion thermodynamics of the transmembrane WALP peptide

    International Nuclear Information System (INIS)

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA)n (L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide’s insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum—in contrast to both atomistic simulations and the alternative PLUM force field. Even though the two coarse-grained models reproduce the free energy of insertion of individual amino acids side chains, they both underestimate its corresponding value for the full peptide (as compared with atomistic simulations), hinting at cooperative physics beyond the residue level. Folding of WALP in the two environments indicates the helix as the most stable structure, though with different relative stabilities and chain-length dependence

  12. Root cause of incomplete control rod insertions at Westinghouse reactors

    International Nuclear Information System (INIS)

    Within the past year, incomplete RCCA insertions have been observed on high burnup fuel assemblies at two Westinghouse PWRs. Initial tests at the Wolf Creek site indicated that the direct cause of the incomplete insertions observed at Wolf Creek was excessive fuel assembly thimble tube distortion. Westinghouse committed to the NRC to perform a root cause analysis by the end of August, 1996. The root cause analysis process used by Westinghouse included testing at ten sites to obtain drag, growth and other characteristics of high burnup fuel assemblies. It also included testing at the Westinghouse hot cell of two of the Wolf Creek incomplete insertion assemblies. A mechanical model was developed to calculate the response of fuel assemblies when subjected to compressive loads. Detailed manufacturing reviews were conducted to determine if this was a manufacturing related issue. In addition, a review of available worldwide experience was performed. Based on the above, it was concluded that the thimble tube distortion observed on the Wolf Creek incomplete insertion assemblies was caused by unusual fuel assembly growth over and above what would typically be expected as a result of irradiation exposure. It was determined that the unusual growth component is a combination of growth due to oxide accumulation and accelerated growth, and would only be expected in high temperature plants on fuel assemblies that see long residence times and high power duties

  13. Computer Modeling of Protocellular Functions: Peptide Insertion in Membranes

    Science.gov (United States)

    Rodriquez-Gomez, D.; Darve, E.; Pohorille, A.

    2006-01-01

    Lipid vesicles became the precursors to protocells by acquiring the capabilities needed to survive and reproduce. These include transport of ions, nutrients and waste products across cell walls and capture of energy and its conversion into a chemically usable form. In modem organisms these functions are carried out by membrane-bound proteins (about 30% of the genome codes for this kind of proteins). A number of properties of alpha-helical peptides suggest that their associations are excellent candidates for protobiological precursors of proteins. In particular, some simple a-helical peptides can aggregate spontaneously and form functional channels. This process can be described conceptually by a three-step thermodynamic cycle: 1 - folding of helices at the water-membrane interface, 2 - helix insertion into the lipid bilayer and 3 - specific interactions of these helices that result in functional tertiary structures. Although a crucial step, helix insertion has not been adequately studied because of the insolubility and aggregation of hydrophobic peptides. In this work, we use computer simulation methods (Molecular Dynamics) to characterize the energetics of helix insertion and we discuss its importance in an evolutionary context. Specifically, helices could self-assemble only if their interactions were sufficiently strong to compensate the unfavorable Free Energy of insertion of individual helices into membranes, providing a selection mechanism for protobiological evolution.

  14. Folding and insertion thermodynamics of the transmembrane WALP peptide

    Energy Technology Data Exchange (ETDEWEB)

    Bereau, Tristan, E-mail: bereau@mpip-mainz.mpg.de [Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz (Germany); Bennett, W. F. Drew [Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1 (Canada); Pfaendtner, Jim [Department of Chemical Engineering, University of Washington, Seattle, Washington 98195 (United States); Deserno, Markus [Department of Physics, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213 (United States); Karttunen, Mikko [Department of Mathematics and Computer Science & Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, MetaForum, 5600 MB Eindhoven (Netherlands)

    2015-12-28

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA){sub n} (L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide’s insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum—in contrast to both atomistic simulations and the alternative PLUM force field. Even though the two coarse-grained models reproduce the free energy of insertion of individual amino acids side chains, they both underestimate its corresponding value for the full peptide (as compared with atomistic simulations), hinting at cooperative physics beyond the residue level. Folding of WALP in the two environments indicates the helix as the most stable structure, though with different relative stabilities and chain-length dependence.

  15. Preliminary Analysis on Reactivity Insertion Transient of Natural Circulation Reactor

    International Nuclear Information System (INIS)

    When a malfunction of the reactor control system occurs, there's a chance that the positive reactivity is inserted into the core, resulting in the increase of the core power. With the combination of the failure of the related safety features, this may raise the temperature of the core material beyond the design limit to break its integrity. For the fast nuclear reactors like FFTF and CRBRP, the overpower trip is initiated when the power reaches 115% of rated value to keep the fuel from melting. In this study, the system response to the reactivity insertion transient on a liquid metal cooled natural circulation reactor is analyzed utilizing an in-house code based on a momentum integral model. Utilizing an in-house system analysis code, a set of numerical simulation is carried out on the reactivity ramp insertion transients which showed that the role of reactivity feedback is significant in mitigating the time to failure, and the evolution of the natural circulation mass flow is rather slow to generate meaningful feedback effect. It is also observed that in terms of the peak cladding temperature, smaller reactivity insertion transient generated more severe outcome owing to increased accumulation of the thermal energy within fuel pins. Thus, it may require an extra attention to carefully monitor and capture the mild transients to avoid potential drastic results

  16. ArcGIS Tool: Inserts file name into attribute table

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This ArcGIS model inserts a file name into a feature class attribute table. The tool allows an user to identify features by a field that reference the name of the...

  17. Insertional mutagenesis using Tnt1 retrotransposon in potato

    Science.gov (United States)

    Potato is the third most important food crop in the world. However, genetics and genomics research of potato has lagged behind many major crop species due to its autotetraploidy and a highly heterogeneous genome. Insertional mutagenesis using T-DNA or transposable elements, which is available in sev...

  18. Choice of Tevatron tune with matched low-β insertions

    International Nuclear Information System (INIS)

    The Tevatron lattice, following closely the Main Ring lattice, is made up of approximately 90 FODO cells with six straight sections equally spaced. The straight sections are identical (neglecting the special high-β straight section used for resonant extraction) providing six-fold symmetry. Each straight section is comprised mainly of a pair of doublets, one at each end, in an antisymmetric configuration. The β-functions are matched to the building block cells but the dispersion function is not. The β-function matching implies that the structure is not sensitive to the tune of the sextant (i.e., structure resonances); however, the dispersion mismatch excites off-momentum dipole resonances and the dispersion structure of the ring is therefore sensitive to sextant tunes near an integer. The introduction of low-β insertions can be accomplished by replacing a standard insertion (expanded somewhat from the doublet pair straight section) with a low-β insertion. By matching the ends of the replacement units with respect to β-functions and dispersion function, the full lattice is made insensitive to linear structure resonances. Procedures are described here for introducing into the idealized Tevatron lattice considered here a single matched low-β insertion at BO, and two matched low-β insertions at BO and DO. The main consequence is the need for a significant number of independently powered quadrupoles. Another conclusion is related to the choice of tune, where it is found that the arbitrary rule of maintaining a fixed tune for all configurations should be modified

  19. Multiple Binary Images Watermarking in Spatial and Frequency Domains

    Directory of Open Access Journals (Sweden)

    K.Ganesan

    2012-12-01

    Full Text Available Editing, reproduction and distribution of the digital multimedia are becoming extremely easier and faster with the existence of the internet and the availability of pervasive and powerful multimedia tools. Digital watermarking has emerged as a possible method to tackle these issues. This paper proposes a scheme using which more data can be inserted into an image in different domains using different techniques. This increases the embedding capacity. Using the proposed scheme 24 binary images can be embedded in the DCT domain and 12 binary images can be embedded in the spatial domain using LSB substitution technique in a single RGB image. The proposed scheme also provides an extra level of security to the watermark image by scrambling the image before embedding it into the host image. Experimental results show that the proposed watermarking method results in almost invisible difference between the watermarked image and the original image and is also robust against various image processing attacks.

  20. Multiple Binary Images Watermarking in Spatial and Frequency Domains

    Directory of Open Access Journals (Sweden)

    K.Ganesan

    2011-02-01

    Full Text Available Editing, reproduction and distribution of the digital multimedia are becoming extremely easier and fasterwith the existence of the internet and the availability of pervasive and powerful multimedia tools. Digitalwatermarking has emerged as a possible method to tackle these issues. This paper proposes a schemeusing which more data can be inserted into an image in different domains using different techniques. Thisincreases the embedding capacity. Using the proposed scheme 24 binary images can be embedded in theDCT domain and 12 binary images can be embedded in the spatial domain using LSB substitutiontechnique in a single RGB image. The proposed scheme also provides an extra level of security to thewatermark image by scrambling the image before embedding it into the host image. Experimental resultsshow that the proposed watermarking method results in almost invisible difference between thewatermarked image and the original image and is also robust against various image processing attacks.

  1. Axion domain wall baryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Daido, Ryuji; Kitajima, Naoya [Department of Physics, Tohoku University,Sendai 980-8578 (Japan); Takahashi, Fuminobu [Department of Physics, Tohoku University,Sendai 980-8578 (Japan); Kavli IPMU, TODIAS, University of Tokyo,Kashiwa 277-8583 (Japan)

    2015-07-28

    We propose a new scenario of baryogenesis, in which annihilation of axion domain walls generates a sizable baryon asymmetry. Successful baryogenesis is possible for a wide range of the axion mass and decay constant, m≃10{sup 8}–10{sup 13} GeV and f≃10{sup 13}–10{sup 16} GeV. Baryonic isocurvature perturbations are significantly suppressed in our model, in contrast to various spontaneous baryogenesis scenarios in the slow-roll regime. In particular, the axion domain wall baryogenesis is consistent with high-scale inflation which generates a large tensor-to-scalar ratio within the reach of future CMB B-mode experiments. We also discuss the gravitational waves produced by the domain wall annihilation and its implications for the future gravitational wave experiments.

  2. Domain Theory for Concurrency

    DEFF Research Database (Denmark)

    Nygaard, Mikkel

    Concurrent computation can be given an abstract mathematical treatment very similar to that provided for sequential computation by domain theory and denotational semantics of Scott and Strachey. A simple domain theory for concurrency is presented. Based on a categorical model of linear logic and...... nondeterministic dataflow. The domain theory can be generalised to presheaf models, providing a more refined treatment of nondeterministic branching and supporting notions of bisimulation. The operational semantics for HOPLA is guided by the idea that derivations of transitions in the operational semantics should...... correspond to elements of the presheaf denotations. Similar guidelines lead to an operational semantics for the first-order fragment of Affine HOPLA. An extension of the operational semantics to the full language is based on a stable denotational semantics which associates to each computation the minimal...

  3. Large domain graphene

    OpenAIRE

    Li, Xuesong; Magnuson, Carl W.; Venugopal, Archana; Vogel, Eric M.; Rodney S. Ruoff; Colombo, Luigi

    2010-01-01

    Graphene growth by chemical vapor deposition has received a lot of attention recently owing to the ease with which large area films can be grown, but growth of large domain or equivalently large grain size has not been reported yet. In this brevia, we report on a CVD process that yields graphene with domains of hundreds of micrometers, by very low pressure CVD, less than 50 mTorr, and very low precursor flow rates using methane as the source of carbon on the inside of copper foil enclosures a...

  4. The Insertion of Fluorescent Proteins in a Variable Region of Respiratory Syncytial Virus L Polymerase Results in Fluorescent and Functional Enzymes But with Reduced Activities

    OpenAIRE

    Fix, Jenna; Galloux, Marie; Blondot, Marie-Lise; Eléouët, Jean-François

    2011-01-01

    The respiratory syncytial virus (RSV) Large protein L is the catalytic subunit of the RNA-dependent RNA polymerase complex. Currently, no structural information is available for RSV L. Sequence alignments of L protein from human and bovine strains of RSV revealed the existence of two variable regions, VR1 and VR2. Following comparison with morbillivirus and rhabdovirus L genes, VR2, which is located between domains V and VI, was chosen as an insertion site for sequences encoding the epitope t...

  5. Atomic Pseudo-Valuation Domains

    CERN Document Server

    Stines, Elijah

    2012-01-01

    Pseudo-valuation domains have been studied since their introduction in 1978 by Hedstrom and Houston. Related objects, boundary valuation domains, were introduced by Maney in 2004. Here, it is shown that the class of atomic pseudo-valuation domains coincides with the class of boundary valuation domains. It is also shown that power series rings and generalized power series rings give examples of pseudo-valuation domains whose congruence lattices can be characterized. The paper also introduces, and makes use of, a sufficient condition on the group of divisibility of a domain to guarantee that it is a pseudo-valuation domain.

  6. Domain: Labour market

    NARCIS (Netherlands)

    Oude Mulders, J.; Wadensjö, E.; Hasselhorn, H.M.; Apt, W.

    2015-01-01

    This domain chapter is dedicated to summarize research on the effects of labour market contextual factors on labour market participation of older workers (aged 50+) and identify research gaps. While employment participation and the timing of (early) retirement is often modelled as an individual deci

  7. Skyrmions and domain walls

    OpenAIRE

    Piette, B.; Zakrzewski, W. J.

    1997-01-01

    We study the 3+1 dimensional Skyrme model with a mass term different from the usual one. We show that this new model possesses domain walls solutions. We describe how, in the equivalent 2+1 dimensional model, the Skyrmion is absorbed by the wall.

  8. On-line air-tightness and insertion loss simultaneous detection method of high air-tightness fiber optic penetration connector

    Science.gov (United States)

    Zhang, Jingchuan; Yang, Xiaoning; Wang, Jing; Jiang, Junfeng

    2015-08-01

    The high air-tightness multicore fiber optic penetration connector is a core component for the optical fiber sensing and communication technologies applied in the space environment simulator under the vacuum thermal environment. High air-tightness and insertion loss are the two key indexes of the fiber optic penetration connector. The air-tightness and insertion loss on-line synchronous detection method was proposed. First, established hardware-in-the-loop testing platform by using the vacuum pumping system, the vacuum vessel, the helium mass spectrometer leak detector and optical time-domain reflectmeter, then, described the air tightness and insertion loss on-line detection principle, finally, designed a detection test scheme and air-tightness and insertion loss were tested. Experimental results indicate that the leakage rate is lower than 1.0×10-7Pa•L/S, the minimum of which is1.0×10-10Pa•L/S and the insertion loss at wave length window 1550 nm is +/-0.07db, which is less than +/-0.1db. It can lay the data basis for the design of opto-mechanical combination and later period fine processing.

  9. Real-time RT-PCR expression analysis of chitinase and endoglucanase genes in the three-way interaction between the biocontrol strain Clonostachys rosea IK726, Botrytis cinera and strawberry

    DEFF Research Database (Denmark)

    Mamarabadi, Mojtaba; Jensen, Birgit; Jensen, Søren Dan Funck;

    2008-01-01

    Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two endoglucan......Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two...... endoglucanase genes from C. rosea strain IK726 was analyzed using real-time RT-PCR in vitro and in strawberry leaves during interaction with B. cinerea. Specific primers were designed for ß-tubulin genes from C. rosea and B. cinerea, respectively, and a gene encoding a DNA-binding protein (DBP) from strawberry......, allowing in situ activity assessment of each fungus in vitro and during their interaction on strawberry leaves. Growth of B. cinerea was inhibited in all pathogen-antagonist interactions while the activity of IK726 was slightly increased. In all in vitro interactions, four of the six genes were upregulated...

  10. A search for RNA insertions and NS3 gene duplication in the genome of cytopathic isolates of bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    V.L. Quadros

    2006-07-01

    Full Text Available Calves born persistently infected with non-cytopathic bovine viral diarrhea virus (ncpBVDV frequently develop a fatal gastroenteric illness called mucosal disease. Both the original virus (ncpBVDV and an antigenically identical but cytopathic virus (cpBVDV can be isolated from animals affected by mucosal disease. Cytopathic BVDVs originate from their ncp counterparts by diverse genetic mechanisms, all leading to the expression of the non-structural polypeptide NS3 as a discrete protein. In contrast, ncpBVDVs express only the large precursor polypeptide, NS2-3, which contains the NS3 sequence within its carboxy-terminal half. We report here the investigation of the mechanism leading to NS3 expression in 41 cpBVDV isolates. An RT-PCR strategy was employed to detect RNA insertions within the NS2-3 gene and/or duplication of the NS3 gene, two common mechanisms of NS3 expression. RT-PCR amplification revealed insertions in the NS2-3 gene of three cp isolates, with the inserts being similar in size to that present in the cpBVDV NADL strain. Sequencing of one such insert revealed a 296-nucleotide sequence with a central core of 270 nucleotides coding for an amino acid sequence highly homologous (98% to the NADL insert, a sequence corresponding to part of the cellular J-Domain gene. One cpBVDV isolate contained a duplication of the NS3 gene downstream from the original locus. In contrast, no detectable NS2-3 insertions or NS3 gene duplications were observed in the genome of 37 cp isolates. These results demonstrate that processing of NS2-3 without bulk mRNA insertions or NS3 gene duplications seems to be a frequent mechanism leading to NS3 expression and BVDV cytopathology.

  11. Fuzzy Bases of Fuzzy Domains

    OpenAIRE

    Sanping Rao; Qingguo Li

    2013-01-01

    This paper is an attempt to develop quantitative domain theory over frames. Firstly, we propose the notion of a fuzzy basis, and several equivalent characterizations of fuzzy bases are obtained. Furthermore, the concept of a fuzzy algebraic domain is introduced, and a relationship between fuzzy algebraic domains and fuzzy domains is discussed from the viewpoint of fuzzy basis. We finally give an application of fuzzy bases, where the image of a fuzzy domain can be preserved under some special ...

  12. Atomic Pseudo-Valuation Domains

    OpenAIRE

    Stines, Elijah

    2012-01-01

    Pseudo-valuation domains have been studied since their introduction in 1978 by Hedstrom and Houston. Related objects, boundary valuation domains, were introduced by Maney in 2004. Here, it is shown that the class of atomic pseudo-valuation domains coincides with the class of boundary valuation domains. It is also shown that power series rings and generalized power series rings give examples of pseudo-valuation domains whose congruence lattices can be characterized. The paper also introduces, ...

  13. Cross-Domain Sparse Coding

    OpenAIRE

    Wang, Jim Jing-Yan

    2013-01-01

    Sparse coding has shown its power as an effective data representation method. However, up to now, all the sparse coding approaches are limited within the single domain learning problem. In this paper, we extend the sparse coding to cross domain learning problem, which tries to learn from a source domain to a target domain with significant different distribution. We impose the Maximum Mean Discrepancy (MMD) criterion to reduce the cross-domain distribution difference of sparse codes, and also ...

  14. Superconformal Ward Identities for Green Functions with Multiple Supercurrent Insertions

    CERN Document Server

    Erdmenger, J; Erdmenger, Johanna; Rupp, Christian

    1999-01-01

    Superconformal Ward identities for N=1 supersymmetric quantum field theories in four dimensions are convenienty obtained in the superfield formalism by combining diffeomorphisms and Weyl transformations on curved superspace. Using this approach we study the superconformal transformation properties of Green functions with one or more insertions of the supercurrent to all orders in perturbation theory. For the case of two insertions we pay particular attention to fixing the additional counterterms present, as well as to the purely geometrical anomalies which contribute to the transformation behaviour. Moreover we show in a scheme-independent way how the quasi-local terms in the Ward identities are related to similar terms which contribute to the supercurrent two and three point functions. Furthermore we relate our superfield approach to similar studies which use the component formalism by discussing the implications of our approach for the components of the supercurrent and of the supergravity prepotentials.

  15. Calculation of magnetic error fields in hybrid insertion devices

    International Nuclear Information System (INIS)

    The Advanced Light Source (ALS) at the Lawrence Berkeley Laboratory requires insertion devices with fields sufficiently accurate to take advantage of the small emittance of the ALS electron beam. To maintain the spectral performance of the synchrotron radiation and to limit steering effects on the electron beam these errors must be smaller than 0.25%. This paper develops a procedure for calculating the steering error due to misalignment of the easy axis of the permanent magnet material. The procedure is based on a three dimensional theory of the design of hybrid insertion devices developed by one of us. The acceptable tolerance for easy axis misalignment is found for a 5 cm period undulator proposed for the ALS. 11 refs., 5 figs

  16. Transients analysis by reactivity insertion in research reactors

    International Nuclear Information System (INIS)

    PARET code was used to simulate accidental situations arising from positive reactivity insertions, in order to analyze the behavior of RP-10 reactor. The simulations considered three different cases: First is for the reactor operating at 10 Mw nominal power with 3 pumps in use, the second, at 6.6 Mw with only one pump working. In all cases the reactor trip was assumed when a 12 Mw power level is reached. An additional simulation for the reactor operating at 50w before the reactivity insertion, showed to be the worst accidental situation of all cases because of the higher temperature and power rise. Hot channel thermohydraulic and kinetic parameters have been evaluated at each axial mesh point and transient time step. None of the cases showed melting of fuel plates

  17. Construction of targeted insertion mutations in Francisella tularensis subsp. novicida.

    Science.gov (United States)

    Liu, Jirong; Zogaj, Xhavit; Barker, Jeffrey R; Klose, Karl E

    2007-10-01

    Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance cassettes and splicing by overlap extension (SOE). This process enables fast and efficient construction of targeted insertion mutations in F. tularensis subsp. novicida that have characteristics of nonpolar mutations; optimized targeted mutagenesis strategies will promote the study of this mysterious bacterium and facilitate vaccine development against tularemia. Moreover the general strategy of gene disruption by PCR-based antibiotic resistance cassette insertion is broadly applicable to many bacterial species. PMID:18019340

  18. Evaluation of coated fuel particle temperature during reactivity insertion events

    International Nuclear Information System (INIS)

    In the High Temperature Gas-cooled Reactors (HTGRs), no adiabatic increase of fuel temperature occurs unless extremely large reactivity is added within a short time even at abnormal condition. Then, in the acceptance criteria of the High Temperature Engineering Test Reactor (HTTR), the maximum fuel temperature is restricted instead of the fuel enthalpy. As one of the studies of fuel behavior under abnormal conditions, temperature transient in coated fuel particle is calculated during reactivity insertion events, and the following results are obtained. (1) Since there is thermal diffusion from fuel kernel to coating layers, the fuel kernel temperature does not increase excessively even under the event of large reactivity insertion rate such as rapid control rod withdrawal accident of HTTR. (2) The maximum fuel kernel temperature is affected by the gap width between the buffer and the inner PyC layers, the composition of gas contained in the gap and the surface area of gap. (author)

  19. Challenges for Insertion of Structural Nanomaterials in Aerospace Applications

    Science.gov (United States)

    Sochi, Emilie J.

    2012-01-01

    In the two decades since Iijima's report on carbon nanotubes (CNT), there has been great interest in realizing the benefits of mechanical properties observed at the nanoscale in large-scale structures. The weight savings possible due to dramatic improvements in mechanical properties relative to state-of-the-art material systems can be game changing for applications like aerospace vehicles. While there has been significant progress in commercial production of CNTs, major aerospace applications that take advantage of properties offered by this material have yet to be realized. This paper provides a perspective on the technical challenges and barriers for insertion of CNTs as an emerging material technology in aerospace applications and proposes approaches that may reduce the typical timeframe for technology maturation and insertion into aerospace structures.

  20. Research of Insertion Mechanism of Flexible Search and Rescue Robot

    Directory of Open Access Journals (Sweden)

    Shen Linyong

    2015-01-01

    Full Text Available Due to various factors, the global scope disaster events occur inevitably every year. Terrorism, natural disasters or destructive earthquakes tend to cause a large number of people buried in the ruins of buildings. The research of search and rescue instruments research is mainly focused on snake-like robot and life detector both at home and abroad. The endoscopic search and rescue robot in this paper combines the characteristics of these two kinds of search and rescue equipment. This thesis mainly studies the insertion mechanism of endoscopic flexible search and rescue robot. Based on the mechanics characteristics of the flexible robot body, this paper analyzes several common failure modes during the insertion into the ruins. This article puts forward the scheme of segmented gradual pulling-pushing, which plays an important role in promoting search and rescue work.

  1. Operation of synchrotron light sources with multiple insertion devices

    International Nuclear Information System (INIS)

    The stability requirements of the next generation of synchrotron radiation facilities have been achieved on insertion device beamlines of existing rings. However, one insertion device (ID) affects the stored beam and hence the performance of all the other beamlines. Since the effects of the undulators are cumulative, higher levels of performance are required of the accelerator in order to meet and exceed present day standards in rings with many undulators. This paper will report experience to date in the areas mentioned above at several multi-undulator facilities and efforts to address these problems at facilities in the planning and construction phase. Section II will treat orbit control and feedback. Section III will describe work on linear and nonlinear effects of ideal undulators in accelerators. Section IV will mention undulator imperfections and the demands they make on the accelerator control system

  2. CUDA accelerated simulation of needle insertions in deformable tissue

    Science.gov (United States)

    Patriciu, Alexandru

    2012-10-01

    This paper presents a stiff needle-deformable tissue interaction model. The model uses a mesh-less discretization of continuum; avoiding thus the expensive remeshing required by the finite element models. The proposed model can accommodate both linear and nonlinear material characteristics. The needle-deformable tissue interaction is modeled through fundamental boundaries. The forces applied by the needle on the tissue are divided in tangent forces and constraint forces. The constraint forces are adaptively computed such that the material is properly constrained by the needle. The implementation is accelerated using NVidia CUDA. We present detailed analysis of the execution timing in both serial and parallel case. The proposed needle insertion model was integrated in a custom software that loads DICOM images, generate the deformable model, and can simulate different insertion strategies.

  3. First two barrel ECAL supermodules inserted in CMS HCAL

    CERN Multimedia

    K.Bell

    2006-01-01

    The first two barrel "supermodules" for the CMS Electromagnetic Calorimeter (ECAL) have been inserted into the barrel hadron calorimeter (HCAL) in the experimental hall (called SX5) in Cessy in preparation for the forthcoming magnet test and cosmic challenge (MTCC). Each of the two supermodules contains 1700 lead tungstate crystals in glass-fibre alveolar support structures, with associated avalanche photodiodes (APDs, for scintillation light detection), electronics and cooling system. The barrel ECAL will consist of 36 supermodules, many of which have already been produced (see CERN Bulletin 17-18, 2006). Team from CMS ECAL, CMS Integration and CEA-DAPNIA were involved in the insertion, with the production/integration of the supermodules themselves involving many technicians, engineers and physicists from many institutes. From left to right: Olivier Teller, Maf Alidra and Lucien Veillet.

  4. Description of popular Science texts inserted in biology textbooks

    Directory of Open Access Journals (Sweden)

    Pedro Henrique Ribeiro de Souza

    2015-08-01

    Full Text Available Popularisation of Science texts are used in Science Education for contextualizing the curriculum content and relating it to the daily lives of students. Textbooks insert popular science texts, mostly adapted, which can interfere in its original content. This work identified the popular science texts in nine teaching collections of high school Biology, approved by PNLD/2015. 398 texts were found, highlighting the collection of Favaretto. Most of them were adapted and were in Ecology, Zoology and Evolution chapters. The books used texts from various sources, but stand out newspapers and popular science magazines, such as Ciência Hoje, which indicates the choice of more accessible texts to the students language. It is important to analyze these texts and understand what changes were made in inserting them in textbooks.

  5. Insertion devices for the Advanced Light Source at LBL

    International Nuclear Information System (INIS)

    The Advanced Light Source (ALS) at the Lawrence Berkeley Laboratory will be the first of the new generation of dedicated synchrotron light sources to be put into operation. Specially designed insertion devices will be required to realize the high brightness photon beams made possible by the low emittance of the electron beam. The complement of insertion devices on the ALS will include undulators with periods as short as 3.9 cm and one or more high field wigglers. The first device to be designed is a 5 m long, 5 cm period, hybrid undulator. The goal of very high brightness and high harmonic output imposes unusually tight tolerances on the magnetic field quality and thus on the mechanical structure. The design process, using a generic structure for all undulators, is described. 5 refs., 4 figs., 1 tab

  6. Graph Grammars, Insertion Lie Algebras, and Quantum Field Theory

    OpenAIRE

    Marcolli, Matilde; Port, Alexander

    2015-01-01

    Graph grammars extend the theory of formal languages in order to model distributed parallelism in theoretical computer science. We show here that to certain classes of context-free and context-sensitive graph grammars one can associate a Lie algebra, whose structure is reminiscent of the insertion Lie algebras of quantum field theory. We also show that the Feynman graphs of quantum field theories are graph languages generated by a theory dependent graph grammar.

  7. Tumor track seeding: A new complication of fiducial marker insertion

    OpenAIRE

    Zeal Patel, MD; Michele Retrouvey, MD; Harlan Vingan, MD; Scott Williams, MD

    2015-01-01

    In the United States, lung cancer is the leading cause of cancer-related death. Candidates for tumor ablation using CyberKnife® require fiducial placement in or near the target tumor to achieve precision. Placing these reference points may lead to complications including pneumothorax and/or hemorrhage. We report a new complication: the appearance of metastatic foci along the track of the fiducial marker. Since the marker was inserted by traversing the original primary tumor, we hypothesize th...

  8. Insertion DNA Accelerates Meiotic Interchromosomal Recombination in Arabidopsis thaliana.

    Science.gov (United States)

    Sun, Xiao-Qin; Li, Ding-Hong; Xue, Jia-Yu; Yang, Si-Hai; Zhang, Yan-Mei; Li, Mi-Mi; Hang, Yue-Yu

    2016-08-01

    Nucleotide insertions/deletions are ubiquitous in eukaryotic genomes, and the resulting hemizygous (unpaired) DNA has significant, heritable effects on adjacent DNA. However, little is known about the genetic behavior of insertion DNA. Here, we describe a binary transgenic system to study the behavior of insertion DNA during meiosis. Transgenic Arabidopsis lines were generated to carry two different defective reporter genes on nonhomologous chromosomes, designated as "recipient" and "donor" lines. Double hemizygous plants (harboring unpaired DNA) were produced by crossing between the recipient and the donor, and double homozygous lines (harboring paired DNA) via self-pollination. The transfer of the donor's unmutated sequence to the recipient generated a functional β-glucuronidase gene, which could be visualized by histochemical staining and corroborated by polymerase chain reaction amplification and sequencing. More than 673 million seedlings were screened, and the results showed that meiotic ectopic recombination in the hemizygous lines occurred at a frequency  >6.49-fold higher than that in the homozygous lines. Gene conversion might have been exclusively or predominantly responsible for the gene correction events. The direct measurement of ectopic recombination events provided evidence that an insertion, in the absence of an allelic counterpart, could scan the entire genome for homologous counterparts with which to pair. Furthermore, the unpaired (hemizygous) architectures could accelerate ectopic recombination between itself and interchromosomal counterparts. We suggest that the ectopic recombination accelerated by hemizygous architectures may be a general mechanism for interchromosomal recombination through ubiquitously dispersed repeat sequences in plants, ultimately contributing to genetic renovation and eukaryotic evolution. PMID:27189569

  9. Non-harmful insertion of data mimicking computer network attacks

    Science.gov (United States)

    Neil, Joshua Charles; Kent, Alexander; Hash, Jr, Curtis Lee

    2016-06-21

    Non-harmful data mimicking computer network attacks may be inserted in a computer network. Anomalous real network connections may be generated between a plurality of computing systems in the network. Data mimicking an attack may also be generated. The generated data may be transmitted between the plurality of computing systems using the real network connections and measured to determine whether an attack is detected.

  10. Pneumothorax as a complication of central venous catheter insertion

    OpenAIRE

    Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios

    2015-01-01

    The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the “central venous oxygen saturation”), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There...

  11. Accurate Insertion Loss Measurements of the Juno Patch Array Antennas

    Science.gov (United States)

    Chamberlain, Neil; Chen, Jacqueline; Hodges, Richard; Demas, John

    2010-01-01

    This paper describes two independent methods for estimating the insertion loss of patch array antennas that were developed for the Juno Microwave Radiometer instrument. One method is based principally on pattern measurements while the other method is based solely on network analyzer measurements. The methods are accurate to within 0.1 dB for the measured antennas and show good agreement (to within 0.1dB) of separate radiometric measurements.

  12. ISR Superconducting High-Luminosity (low beta ) insertion

    CERN Multimedia

    1981-01-01

    The photograph shows two of the 8 Superconducting Quadrupoles installed in ISR intersection I8 with their helium supply flexible lines,vacuum equipment,power and signal cables. The increase of luminosity produced by this insertion was above a factor 7. On the right one can see part of Open-Axial-Field Magnet. The person on the left side is Stephan Pichler. See also photo 7702690 and its abstract.

  13. On the use of plastic inserts in prestressed railway components.

    OpenAIRE

    Coombs, W.M.

    2016-01-01

    The use of such plastic inserts (such as Vossloh dowels and other soft fastening solutions) in pre-stressed concrete sleepers and crossing bearers has become widespread within the United Kingdom, Europe and the rest of the world. This paper uses a bespoke finite-element analysis tool to, for the first time, investigate the stress state around these plastic inclusions specifically focusing on the likelihood of discontinuity development. Most continuum stress analysis methods are over simplisti...

  14. Strain measurement at the knee ligament insertion sites

    OpenAIRE

    Hinterwimmer, S; Baumgart, R.; Plitz, W.

    2003-01-01

    We describe the modification of an existing method of ligament strain measurement at the knee joint in detail. At ten fresh joint specimens we used that technique where strain gauges are attached to the ligamentous insertions and origins. We both improved the preparation of the attachment site and the application of the strain gauges. In a special apparatus the specimens were moved from 0degrees extension to 100degrees flexion while simulating muscle strength and axial force. Testing was perf...

  15. Ferromagnetic inserts for the ripple correction in NET

    International Nuclear Information System (INIS)

    The possibility of generating large plasmas in NET might be limited by the high values of the toroidal field ripple. A possible means of ripple reduction is to include pieces of ferromagnetic material in the shielding structure. This paper presents the results of 3-D calculations that show how inserts of reasonable size can reduce the ripple to acceptable levels. The influence of the different dimensions of the ferromagnetic sectors is also shown. (author). 2 refs.; 7 figs

  16. Power distribution from insertion device x-ray sources

    International Nuclear Information System (INIS)

    Insertion device (ID) synchrotron x-ray sources on present day and next-generation synchrotron facilities have very attractive spectral properties. In addition however, they are capable of producing x-ray beams with large powers and in some cases, unprecedented power densities. An overview of the spatial and frequency distributions of these sources including the effects of synchrotron particle beam emittance is presented

  17. Insertion of cations into WO3 investigated by QCM techniques

    Czech Academy of Sciences Publication Activity Database

    Vondrák, Jiří; Sedlaříková, M.; Velická, Jana; Špičák, P.; Svoboda, V.; Kazelle, J.

    2007-01-01

    Roč. 11, č. 10 (2007), s. 1459-1462. ISSN 1432-8488 R&D Projects: GA MŽP SN/3/171/05; GA AV ČR(CZ) KJB208130604; GA ČR(CZ) GA104/06/1471 Institutional research plan: CEZ:AV0Z40320502 Keywords : tungsten trioxide * insertion of cations * quartz crystal microbalance Subject RIV: CG - Electrochemistry Impact factor: 1.535, year: 2007

  18. Cytoplasmic Organelle DNA Preferentially Inserts into Open Chromatin

    OpenAIRE

    Wang, Dong; Timmis, Jeremy N.

    2013-01-01

    DNA transfer from chloroplasts and mitochondria to the nucleus is ongoing in eukaryotes but the mechanisms involved are poorly understood. Mitochondrial DNA was observed to integrate into the nuclear genome through DNA double-strand break repair in Nicotiana tabacum. Here, 14 nuclear insertions of chloroplast DNA (nupts) that are unique to Oryza sativa subsp. indica were identified. Comparisons with the preinsertion nuclear loci identified in the related subspecies, O. sativa subsp. japonica,...

  19. The first insertion devices at SSRL - some personal recollections

    Energy Technology Data Exchange (ETDEWEB)

    Winick, H. [Stanford Linear Accelerator Center, CA (United States)

    1995-02-01

    The author recounts his experiences with insertion devices at the Stanford Synchrotron Radiation Laboratory. His first experiences with wigglers occured at the Cambridge Electron Accelerator, and was carried over to SSRL with the proposal for a six pole electromagnetic wiggler. Most modern undulators, and many wigglers are now designed around permanent magnets, and the origin of this transition at SSRL was rather fortuitous and humorous. It reflects some of the personality characteristics of Klaus Halbach.

  20. A training simulator for ultrasound-guided percutaneous nephrostomy insertion

    OpenAIRE

    Rock, B G; Leonard, A P; Freeman, S. J.

    2010-01-01

    Increasing trainee numbers and changes to working patterns have resulted in a scarcity of training opportunities for training-grade doctors wishing to learn nephrostomy tube insertion techniques. A method of introducing trainees to the skills required to perform percutaneous nephrostomy in a safe, non-threatening environment, without risk to patients, is desirable. Commercial and biological nephrostomy phantoms are available, but they are expensive and not widely available, and a cheap, safe,...

  1. Magnetic resonance urography in duplex kidney with ectopic ureteral insertion.

    Science.gov (United States)

    Thambidorai, Conjeevaram Rajendrarao; Anuar, Zulfiqar

    2011-07-01

    This is a report on the use of magnetic resonance urography (MRU) in a 6-year-old girl who presented with urinary incontinence. She had a left duplex kidney with poorly functioning upper moiety and ectopic insertion of the dilated upper pole ureter. MRU has been shown to be superior to conventional imaging techniques in delineating poorly functioning moieties of duplex kidneys and ectopic ureters. PMID:21897576

  2. Magnetic resonance urography in duplex kidney with ectopic ureteral insertion

    OpenAIRE

    Thambidorai, Conjeevaram Rajendrarao; Anuar, Zulfiqar

    2011-01-01

    This is a report on the use of magnetic resonance urography (MRU) in a 6-year-old girl who presented with urinary incontinence. She had a left duplex kidney with poorly functioning upper moiety and ectopic insertion of the dilated upper pole ureter. MRU has been shown to be superior to conventional imaging techniques in delineating poorly functioning moieties of duplex kidneys and ectopic ureters.

  3. Magnetic resonance urography in duplex kidney with ectopic ureteral insertion

    Directory of Open Access Journals (Sweden)

    Conjeevaram Rajendrarao Thambidorai

    2011-01-01

    Full Text Available This is a report on the use of magnetic resonance urography (MRU in a 6-year-old girl who presented with urinary incontinence. She had a left duplex kidney with poorly functioning upper moiety and ectopic insertion of the dilated upper pole ureter. MRU has been shown to be superior to conventional imaging techniques in delineating poorly functioning moieties of duplex kidneys and ectopic ureters.

  4. Description of popular Science texts inserted in biology textbooks

    OpenAIRE

    Pedro Henrique Ribeiro de Souza; Marcelo Borges Rocha

    2015-01-01

    Popularisation of Science texts are used in Science Education for contextualizing the curriculum content and relating it to the daily lives of students. Textbooks insert popular science texts, mostly adapted, which can interfere in its original content. This work identified the popular science texts in nine teaching collections of high school Biology, approved by PNLD/2015. 398 texts were found, highlighting the collection of Favaretto. Most of them were adapted and were in Ecolog...

  5. Soft Tissue Phantoms for Realistic Needle Insertion: A Comparative Study.

    Science.gov (United States)

    Leibinger, Alexander; Forte, Antonio E; Tan, Zhengchu; Oldfield, Matthew J; Beyrau, Frank; Dini, Daniele; Rodriguez Y Baena, Ferdinando

    2016-08-01

    Phantoms are common substitutes for soft tissues in biomechanical research and are usually tuned to match tissue properties using standard testing protocols at small strains. However, the response due to complex tool-tissue interactions can differ depending on the phantom and no comprehensive comparative study has been published to date, which could aid researchers to select suitable materials. In this work, gelatin, a common phantom in literature, and a composite hydrogel developed at Imperial College, were matched for mechanical stiffness to porcine brain, and the interactions during needle insertions within them were analyzed. Specifically, we examined insertion forces for brain and the phantoms; we also measured displacements and strains within the phantoms via a laser-based image correlation technique in combination with fluorescent beads. It is shown that the insertion forces for gelatin and brain agree closely, but that the composite hydrogel better mimics the viscous nature of soft tissue. Both materials match different characteristics of brain, but neither of them is a perfect substitute. Thus, when selecting a phantom material, both the soft tissue properties and the complex tool-tissue interactions arising during tissue manipulation should be taken into consideration. These conclusions are presented in tabular form to aid future selection. PMID:26666228

  6. Folding and insertion thermodynamics of the transmembrane WALP peptide

    CERN Document Server

    Bereau, Tristan; Pfaendtner, Jim; Deserno, Markus; Karttunen, Mikko

    2015-01-01

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA)$_n$(L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide's insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum---in contrast to both atomistic simulations and the alternative PLUM force field. Even tho...

  7. Modeling and characterization of partially inserted electrical connector faults

    Science.gov (United States)

    Tokgöz, ćaǧatay; Dardona, Sameh; Soldner, Nicholas C.; Wheeler, Kevin R.

    2016-03-01

    Faults within electrical connectors are prominent in avionics systems due to improper installation, corrosion, aging, and strained harnesses. These faults usually start off as undetectable with existing inspection techniques and increase in magnitude during the component lifetime. Detection and modeling of these faults are significantly more challenging than hard failures such as open and short circuits. Hence, enabling the capability to locate and characterize the precursors of these faults is critical for timely preventive maintenance and mitigation well before hard failures occur. In this paper, an electrical connector model based on a two-level nonlinear least squares approach is proposed. The connector is first characterized as a transmission line, broken into key components such as the pin, socket, and connector halves. Then, the fact that the resonance frequencies of the connector shift as insertion depth changes from a fully inserted to a barely touching contact is exploited. The model precisely captures these shifts by varying only two length parameters. It is demonstrated that the model accurately characterizes a partially inserted connector.

  8. NURSING CARE IN PATIENTS NEONATES WITH PERIPHERALLY INSERTED CENTRAL CATHETER

    Directory of Open Access Journals (Sweden)

    Anacilda Oliveira Vieira

    2013-12-01

    Full Text Available Introduction: The PICC (peripherally inserted central catheter is a long flexible catheter which is inserted through a peripheral vein, progresses through a needle introducer until the final portion of the vena cava, acquiring characteristics of a central catheter. Objective: To point out the main theoretical and scientific ideas that demonstrate the reliability, competence and ability of nurses to perform the PICC. Methodology: Systematic review of articles, which were found by searching the database scientific journals and bibliographies area. Results: The success of integration depends on the patient assessment and choice of venous access where the catheter will be positioned, and its tip should be in the middle third of the superior vena cava, or the middle third of the inferior vena cava. In neonates, which are used more frequently, proper positioning of the catheter is through nursing care in making the dressing, and the first 24 hours it should be compressive. Ideally, the PICC remains in the vein for periods longer than seven days or until the end of treatment, thus decreasing invasive procedures. Conclusion: According to the Federal Board of Nursing (COFEN, it is lawful for the insertion of PICC nurses, provided it has undergone professional training.

  9. Pneumothorax as a complication of central venous catheter insertion.

    Science.gov (United States)

    Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios; Kuhajda, Ivan; Andjelkovic, Dejan; Zarogoulidis, Konstantinos; Zarogoulidis, Paul

    2015-03-01

    The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the "central venous oxygen saturation"), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There are situations according to the drug administration or length of stay of the catheter that specific systems are indicated such as; a Hickman line, a peripherally inserted central catheter (PICC) line or a Port-a-Cath may be considered because of their smaller infection risk. Sterile technique is highly important here, as a line may serve as a port of entry for pathogenic organisms, and the line itself may become infected with organisms such as Staphylococcus aureus and coagulase-negative Staphylococci. In the current review we will present the complication of pneumothorax after CVC insertion. PMID:25815301

  10. Coaxial needle insertion assistant with enhanced force feedback.

    Science.gov (United States)

    De Lorenzo, Danilo; Koseki, Yoshihiko; De Momi, Elena; Chinzei, Kiyoyuki; Okamura, Allison M

    2013-02-01

    Many medical procedures involving needle insertion into soft tissues, such as anesthesia, biopsy, brachytherapy, and placement of electrodes, are performed without image guidance. In such procedures, haptic detection of changing tissue properties at different depths during needle insertion is important for needle localization and detection of subsurface structures. However, changes in tissue mechanical properties deep inside the tissue are difficult for human operators to sense, because the relatively large friction force between the needle shaft and the surrounding tissue masks the smaller tip forces. A novel robotic coaxial needle insertion assistant, which enhances operator force perception, is presented. This one-degree-of-freedom cable-driven robot provides to the operator a scaled version of the force applied by the needle tip to the tissue, using a novel design and sensors that separate the needle tip force from the shaft friction force. The ability of human operators to use the robot to detect membranes embedded in artificial soft tissue was tested under the conditions of 1) tip force and shaft force feedback, and 2) tip force only feedback. The ratio of successful to unsuccessful membrane detections was significantly higher (up to 50%) when only the needle tip force was provided to the user. PMID:23193302

  11. ANATOMICAL VARIATION OF PALMARIS LONGUS: TENDINOUS ORIGIN AND FLESHY INSERTION

    Directory of Open Access Journals (Sweden)

    Buddhadeb Ghosh

    2015-03-01

    Full Text Available A tendinous origin and fleshy insertion of palmaris longus muscle was observed in the left forearm during routine dissection which was performed on adult male cadaver in the department of Anatomy, Dr. Rajendra Prasad Government Medical College. It was having long tendinous origin from the medial epicondyle of the humerus and the surrounding deep fascia. It was fusiform at the lower middle of the forearm. The fleshy muscular insertion was noted to the flexor retinaculum and few muscular fibers interdigitate with flexor carpi ulnaris muscle and palmar aponeurosis. The length of tendon was 19 inches and fleshy muscular length was 11inches. The median nerve and ulnar nerve was covered by this fleshy insertion. This palmaris longus variation is helpful for the surgeon and the radiologist, orthopaedic, plastic surgeon during any diagnosis of the forearm because this fleshy part of muscle can compress the median nerve and ulnar nerve or it can be mistaken as a tumor or ganglion during radiological or clinical examination.

  12. Non-rotating cementing plug with molded inserts

    International Nuclear Information System (INIS)

    This patent describes an anti-rotation plug set for use with cementing equipment having an insert seat therein, the anti-rotation plug set and the cementing equipment for use in cementing a string of casing into a well bore. It comprises an upper plug including: a non-metallic body member having a plurality of teeth integrally formed on the lower end thereof and an elastomeric covering thereon having, in turn, wipers which engage the interior of the string of casing; and a lower plug including: a non-metallic body member having a bore therethrough, having teeth integrally formed on the upper end thereof which mate with the teeth integrally formed on the lower end of the nonmetallic body member of the upper plug when the upper plug engages the lower plugs, having teeth integrally formed on the lower end thereof, having an insert member in a portion of the bore through the non-metallic body member, and the cementing equipment comprising: an insert seat having teeth thereon which mate with the integrally formed teeth on the lower end of the non-metallic body member of the lower plug of the anti-rotation plug set when the lower plug of the anti-rotation plug set engages the cementing equipment during the cementing of the string of casing into a well bore

  13. Enhancement of Selection, Bubble and Insertion Sorting Algorithm

    Directory of Open Access Journals (Sweden)

    Muhammad Farooq Umar

    2014-07-01

    Full Text Available In everyday life there is a large amount of data to arrange because sorting removes any ambiguities and make the data analysis and data processing very easy, efficient and provides with cost less effort. In this study a set of improved sorting algorithms are proposed which gives better performance and design idea. In this study five new sorting algorithms (Bi-directional Selection Sort, Bi-directional bubble sort, MIDBiDirectional Selection Sort, MIDBidirectional bubble sort and linear insertion sort are presented. Bi-directional Selection Sort and MIDBiDirectional Selection Sort are the enhancement on basic selection sort while Bidirectional bubble sort and MIDBidirectional bubble sort are the enhancement on basic bubble sort by changing the selection and swapping mechanism of data for sorting. Enhanced sorting algorithms reduced the iteration by half and quarter times respectively. Asymptotically complexities of these algorithms are reduced to O (n2/2 and O (n2/4 from O (n2. Linear insertion sort is the enhancement of insertion sort by changing the design of algorithm (convert two loops to one loop. So asymptotically this algorithm is converted to linear time complexity from quadratic complexity. These sorting algorithms are described using C. The proposed algorithms are analyzed using asymptotic analysis and also using machine-running time and compared with their basic sorting algorithms. In this study we also discuss how the performance and complexity can be improved by optimizing the code and design.

  14. Examining Prebiotic Chemistry Using O(^1D) Insertion Reactions

    Science.gov (United States)

    Hays, Brian M.; Laas, Jacob C.; Weaver, Susanna L. Widicus

    2013-06-01

    Aminomethanol, methanediol, and methoxymethanol are all prebiotic molecules expected to form via photo-driven grain surface chemistry in the interstellar medium (ISM). These molecules are expected to be precursors for larger, biologically-relevant molecules in the ISM such as sugars and amino acids. These three molecules have not yet been detected in the ISM because of the lack of available rotational spectra. A high resolution (sub)millimeter spectrometer coupled to a molecular source is being used to study these molecules using O(^1D) insertion reactions. The O(^1D) chemistry is initiated using an excimer laser, and the products of the insertion reactions are adiabatically cooled using a supersonic expansion. Experimental parameters are being optimized by examination of methanol formed from O(^1D) insertion into methane. Theoretical studies of the structure and reaction energies for aminomethanol, methanediol, and methoxymethanol have been conducted to guide the laboratory studies once the methanol experiment has been optimized. The results of the calculations and initial experimental results will be presented.

  15. A Diverse Range of Bacterial and Eukaryotic Chitinases Hydrolyzes the LacNAc (Galβ1-4GlcNAc) and LacdiNAc (GalNAcβ1-4GlcNAc) Motifs Found on Vertebrate and Insect Cells

    DEFF Research Database (Denmark)

    Frederiksen, Rikki F.; Yoshimura, Yayoi; Storgaard, Birgit G.; Paspaliari, Dafni K.; Petersen, Bent O.; Chen, Kowa; Larsen, Tanja; Duus, Jens O.; Ingmer, Hanne; Bovin, Nicolai V.; Westerlind, Ulrika; Blixt, Ola; Palcic, Monica M.; Leisner, Jørgen

    2015-01-01

    There is emerging evidence that chitinases have additional functions beyond degrading environmental chitin, such as involvement in innate and acquired immune responses, tissue remodeling, fibrosis and serving as virulence factors of bacterial pathogens. We have recently shown that both the human...

  16. Reconstitution technology of Charpy surveillance specimens with short insert length

    International Nuclear Information System (INIS)

    As for the shortage of the surveillance specimens to monitor the effect of the irradiation embrittlement of reactor pressure vessels (RPV) materials in case of longer-term operation than present surveillance program of nuclear power plants, the reconstitution of them is considered to be the promising measures. Although the length of the specimen insert is required not less than 18 mm in ASTM E1253-99 which is the technical standard to reconstitute Charpy specimens, the minimum length of the specimen insert required should be 10 mm when L-T direction Charpy specimens that have been applied to the early domestic nuclear power plants are reconstituted into T-L direction specimens in order to test the upper shelf absorbed energy of T-L direction specimens. This paper presents the current status of the research consigned by Ministry of Economy, Trade and Industry (METI) in Japan on the applicability of the reconstituted Charpy specimens with short length of the specimen insert. The length of the specimen insert to preserve the absorbed energy of the Charpy specimen is correlated to the absorbed energy of its material. The significant part of upper shelf energy is attributed to the energy for the plastic deformation zone near V-notch in the Charpy specimen. To preserve the absorbed energy, the anticipated plastically deformed zone shall not be affected by the reconstitution procedure. In order to clarify the condition for preserving the absorbed energy in the case of reconstitution, the preliminary data has been obtained using un-irradiated and irradiated Charpy specimens, and the following results have been obtained by the tests carried out in this research. 1) The plastic deformation widths have been estimated by measuring the hardness distribution near the V-notch of the un-irradiated Charpy impact tested specimens, correlated to the absorbed energy. 2) The absorbed energy shifts of reconstituted, un-irradiated Charpy specimens with various length of the specimen

  17. Test manufacturing of copper canisters with cast inserts. Assessment report

    Energy Technology Data Exchange (ETDEWEB)

    Andersson, C.G

    1998-08-01

    The current design of canisters for the deep repository for spent nuclear fuel consists of an outer corrosion-protective copper casing in the form of a tubular section with lid and bottom and an inner pressure-resistant insert. The insert is designed to be manufactured by casting and inside are channels in which the fuel assemblies are to be placed. Over the last years, a number of full-scale manufacturing tests of all canister components have been carried out. The purpose has been to determine and develop the best manufacturing technique and to establish long-term contacts with the best suppliers of material and technology. Part of the work has involved the developing and implementing of a quality assurance system in accordance with ISO 9001, covering the whole chain from suppliers of material up to and including the delivery of assembled canisters. This report consists of a description of the design of the canister together with current drawings and complementary technical specifications stipulating, among other things, requirements placed on different materials. The different manufacturing methods that have been used are also described and commented on in both text and illustrations. For the manufacturing of copper tubes, the roll-forming of rolled plate to tube halves and longitudinal welding is a method that has been tested on a relatively large number of tubes by now, and that probably can be developed into a functioning production method. However, the very promising outcome of performed tests on seamless tube manufacturing, has resulted in a change in direction in tube manufacturing, focusing on continued testing of extrusion as well as pierce and draw processing in the immediate future. In connection with ongoing operations, new manufacturing tests of tubes with less material thickness will be carried out. Test manufacturing of cast inserts has resulted in the choice of nodular iron as material in the continued work. This improvement in design has resulted

  18. Test manufacturing of copper canisters with cast inserts. Assessment report

    International Nuclear Information System (INIS)

    The current design of canisters for the deep repository for spent nuclear fuel consists of an outer corrosion-protective copper casing in the form of a tubular section with lid and bottom and an inner pressure-resistant insert. The insert is designed to be manufactured by casting and inside are channels in which the fuel assemblies are to be placed. Over the last years, a number of full-scale manufacturing tests of all canister components have been carried out. The purpose has been to determine and develop the best manufacturing technique and to establish long-term contacts with the best suppliers of material and technology. Part of the work has involved the developing and implementing of a quality assurance system in accordance with ISO 9001, covering the whole chain from suppliers of material up to and including the delivery of assembled canisters. This report consists of a description of the design of the canister together with current drawings and complementary technical specifications stipulating, among other things, requirements placed on different materials. The different manufacturing methods that have been used are also described and commented on in both text and illustrations. For the manufacturing of copper tubes, the roll-forming of rolled plate to tube halves and longitudinal welding is a method that has been tested on a relatively large number of tubes by now, and that probably can be developed into a functioning production method. However, the very promising outcome of performed tests on seamless tube manufacturing, has resulted in a change in direction in tube manufacturing, focusing on continued testing of extrusion as well as pierce and draw processing in the immediate future. In connection with ongoing operations, new manufacturing tests of tubes with less material thickness will be carried out. Test manufacturing of cast inserts has resulted in the choice of nodular iron as material in the continued work. This improvement in design has resulted

  19. Time Domain Induced Polarization

    DEFF Research Database (Denmark)

    Fiandaca, Gianluca; Auken, Esben; Christiansen, Anders Vest;

    2012-01-01

    Time-domain-induced polarization has significantly broadened its field of reference during the last decade, from mineral exploration to environmental geophysics, e.g., for clay and peat identification and landfill characterization. Though, insufficient modeling tools have hitherto limited the use...... of time-domaininduced polarization for wider purposes. For these reasons, a new forward code and inversion algorithm have been developed using the full-time decay of the induced polarization response, together with an accurate description of the transmitter waveform and of the receiver transfer...... is above 100%. Furthermore, the presence of low-pass filters in time-domain-induced polarization instruments affects the early times of the acquired decays (typically up to 100 ms) and has to be modeled in the forward response to avoid significant loss of resolution. The developed forward code has...

  20. Domain-specific languages

    OpenAIRE

    Jasný, Vojtěch

    2009-01-01

    The topic of the thesis are domain-specific languages (DSL) and their use in software development. The target audience are developers interested in learning more about this progressive area of software development. It starts with a necessary theoretical introduction to programming languages. Then, a classification of DSLs is given and software development methodologies based on DSLs are described, notably Language Oriented Programming and Intentional Programming. Another important piece in co...

  1. Implementing maritime domain awareness

    OpenAIRE

    Watts, Robert B.

    2006-01-01

    CHDS State/Local As an attempt to gain understanding of everything in the global maritime environment that can impact the security of the United States, the Maritime Domain Awareness initiative is one of the most ambitious projects ever undertaken by the U.S. government. Information that falls under the prevue of MDA is tremendously diverse and complex, having application in the regulatory, law enforcement, and military arenas. As such, MDA is a multi-agency effort that encompasses 16 resp...

  2. Ultrasound-guided central line insertion and standard peripherally inserted catheter placement in preterm infants: Comparing results from prospective study in a single-center

    Directory of Open Access Journals (Sweden)

    Dany Antanios Al Hamod

    2016-01-01

    Full Text Available Background: Among preterm infants, the peripherally inserted central catheter (PICC is the standard line for central venous access; however, its placement exposes them to hypothermia and pain. Ultrasound (US-guided central line insertion may be less morbid than standard PICC line. Aims: To determine the ease, success rate, and morbidity associated with US-guided central line insertion in the internal jugular vein (IJV by comparing it to the standard PICC line placement. Materials and Methods: This is a single-center nonrandomized prospective study evaluating preterm infants between October 2013 and June 2014. Patients were allocated into two groups: The standard group (control group who underwent blind PICC line insertion and the intervention group who underwent a percutaneous US-guided central line insertion in the IJV. The epicutaneo-cava-catheter was used in both groups. Results: Fifty neonates were enrolled on study. A statistically difference in favor of US-IJV insertion was noted concerning the rate of successful first attempt (P < 0.001, insertion (P = 0.001, and procedure duration (P < 0.001 and number of trials (P < 0.001 compared to PICC. No difference in complications (P = 1.000 was noted. Conclusion: US guided catheterization of the IJV technique is faster than PICC line insertion with higher rates of successful first attempt and insertion, less procedure duration and fewer number of trials compared to PICC line insertion. There were no differences in complications.

  3. Sampling and TDR probe insertion in the determination of the volumetric soil water content

    Directory of Open Access Journals (Sweden)

    Teixeira W. G.

    2003-01-01

    Full Text Available Volumetric soil water content (theta can be evaluated in the field by direct or indirect methods. Among the direct, the gravimetric method is regarded as highly reliable and thus often preferred. Its main disadvantages are that sampling and laboratory procedures are labor intensive, and that the method is destructive, which makes resampling of a same point impossible. Recently, the time domain reflectometry (TDR technique has become a widely used indirect, non-destructive method to evaluate theta. In this study, evaluations of the apparent dielectric number of soils (epsilon and samplings for the gravimetrical determination of the volumetric soil water content (thetaGrav were carried out at four sites of a Xanthic Ferralsol in Manaus - Brazil. With the obtained epsilon values, theta was estimated using empirical equations (thetaTDR, and compared with thetaGrav derived from disturbed and undisturbed samples. The main objective of this study was the comparison of thetaTDR estimates of horizontally as well as vertically inserted probes with the thetaGrav values determined by disturbed and undisturbed samples. Results showed that thetaTDR estimates of vertically inserted probes and the average of horizontally measured layers were only slightly and insignificantly different. However, significant differences were found between the thetaTDR estimates of different equations and between disturbed and undisturbed samples in the thetaGrav determinations. The use of the theoretical Knight et al. model, which permits an evaluation of the soil volume assessed by TDR probes, is also discussed. It was concluded that the TDR technique, when properly calibrated, permits in situ, nondestructive measurements of q in Xanthic Ferralsols of similar accuracy as the gravimetric method.

  4. Domain decomposition method for solving elliptic problems in unbounded domains

    International Nuclear Information System (INIS)

    Computational aspects of the box domain decomposition (DD) method for solving boundary value problems in an unbounded domain are discussed. A new variant of the DD-method for elliptic problems in unbounded domains is suggested. It is based on the partitioning of an unbounded domain adapted to the given asymptotic decay of an unknown function at infinity. The comparison of computational expenditures is given for boundary integral method and the suggested DD-algorithm. 29 refs.; 2 figs.; 2 tabs

  5. Comparison of the domain and frequency domain state feedbacks

    International Nuclear Information System (INIS)

    In this paper, we present explicitly the equivalence of the time domain and frequency domain state feedbacks, as well as the dynamic state feedback and a modified frequency domain state feedback, from the closed-loop transfer function point of view. The difference of the two approaches is also shown

  6. On Free Z-domains%自由Z-domain

    Institute of Scientific and Technical Information of China (English)

    姚丽娟; 徐晓泉

    2011-01-01

    给出由偏序集生成的自由Z-domain,讨论自由Z-domain和偏序集的Dedekind-MacNeille完备化的一些性质.%The free Z-domains generated by posets are given.Some properties of tte free Z-domains and the Dedekind-MacNeille completions of posets are investigated.

  7. Insertion of the CMS coil into the barrel yoke on 14 September 2005

    CERN Multimedia

    Maximilien Brice

    2005-01-01

    Insertion of the CMS coil into the barrel yoke on 14 September 2005. The pictures have been taken in the CMS experimental hall SX5 in Cessy, neighbouring France. The second picture shows the insertion of the Inner Vacuum Tank.

  8. Ultrasound-guided Central Line Insertion and Standard Peripherally Inserted Catheter Placement in Preterm Infants: Comparing Results from Prospective Study in a Single-center

    OpenAIRE

    Dany Antanios Al Hamod; Smart Zeidan; Ayah Al Bizri; Georges Baaklini; Yolla Nassif

    2016-01-01

    Background: Among preterm infants, the peripherally inserted central catheter (PICC) is the standard line for central venous access; however, its placement exposes them to hypothermia and pain. Ultrasound (US)-guided central line insertion may be less morbid than standard PICC line. Aims: To determine the ease, success rate, and morbidity associated with US-guided central line insertion in the internal jugular vein (IJV) by comparing it to the standard PICC line placement. Materials and Metho...

  9. Summarization by domain ontology navigation

    DEFF Research Database (Denmark)

    Andreasen, Troels; Bulskov, Henrik

    2013-01-01

    of the subject. In between these two extremes, conceptual summaries encompass selected concepts derived using background knowledge. We address in this paper an approach where conceptual summaries are provided through a conceptualization as given by an ontology. The ontology guiding the summarization can...... be a simple taxonomy or a generative domain ontology. A domain ontology can be provided by a preanalysis of a domain corpus and can be used to condense improved summaries that better reflects the conceptualization of a given domain....

  10. Domain Engineering: An Empirical Study

    OpenAIRE

    Harris, Charles; Frakes, William B.

    2006-01-01

    This paper presents a summary and analysis of data gathered from thirteen domain engineering projects, participant surveys, and demographic information. Taking a failure modes approach, project data is compared to an ideal model of the DARE methodology, revealing valuable insights into points of failure in the domain engineering process. This study suggests that success is a function of the domain analyst’s command of a specific set of domain engineering concepts and skills, the time invested...

  11. A Convenient Category of Domains

    OpenAIRE

    Battenfeld, Ingo; Schröder, Matthias; Simpson, Alexander

    2007-01-01

    We motivate and define a category of topological domains, whose objects are certain topological spaces, generalising the usual ω-continuous dcppos of domain theory. Our category supports all the standard constructions of domain theory, including the solution of recursive domain equations. It also supports the construction of free algebras for (in)equational theories, can be used as the basis for a theory of computability, and provides a model of parametric polymorphism.

  12. A convenient category of domains

    OpenAIRE

    Battenfeld, Ingo; Schröder, Matthias; Simpson, Alex

    2007-01-01

    We motivate and define a category of "topological domains", whose objects are certain topological spaces, generalising the usual $omega$-continuous dcppos of domain theory. Our category supports all the standard constructions of domain theory, including the solution of recursive domain equations. It also supports the construction of free algebras for (in)equational theories, provides a model of parametric polymorphism, and can be used as the basis for a theory of computab...

  13. External ear canal cholesteatoma after ventilation tube insertion and mastoidectomy

    Directory of Open Access Journals (Sweden)

    Đerić Dragoslava

    2012-01-01

    Full Text Available Introduction. Etiopathogenetically, there are two types of chollesteatomas: congenital, and acquired. Numerous theories in the literature try to explain the nature of the disease, however, the question about cholesteatomas remain still unanswered. The aim of the study was to present a case of external ear canal cholesteatoma (EEC developed following microsurgery (ventilation tube insertion and mastoidectomy, as well as to point ant possible mechanisms if its development. Case report. A 16-yearold boy presented a 4-month sense of fullness in the ear and otalgia on the left side. A year before, mastoidectomy and posterior atticotomy were performed with ventilation tube placement due to acute purulent mastoiditis. Diagnosis was based on otoscopy examination, audiology and computed tomography (CT findings. CT showed an obliterative soft-tissue mass completely filled the external ear canal with associated erosion of subjacent the bone. There were squamous epithelial links between the canal cholesteatoma and lateral tympanic membrane surface. They originated from the margins of tympanic membrane incision made for a ventilation tube (VT insertion. The position of VT was good as well as the aeration of the middle ear cavity. The tympanic membrane was intact and of normal appearance without middle ear extension or mastoid involvement of cholesteatoma. Cholesteatoma and ventilation tube were both removed. The patient recovered without complications and shortly audiology revealed hearing improving. Follow-up 2 years later, however, showed no signs of the disease. Conclusion. There could be more than one potential delicate mechanism of developing EEC in the ear with VT insertion and mastoidectomy. It is necessary to perform routine otologic surveillance in all patients with tubes. Affected ear CT scan is very helpful in showing the extent of cholesteatoma and bony defects, which could not be assessed by otoscopic examination alone.

  14. Estudio de los genes allinasa y quitinasa en el ajo costarricense (Allium sativum L. Study of the genes alliinase and chitinase in materials of costarican garlic (Allium sativum L

    Directory of Open Access Journals (Sweden)

    Karina Barboza Rojas

    2013-03-01

    Full Text Available El cultivo del ajo en Costa Rica se ha visto afectado por la calidad y cantidad de semillas almacenadas. La producción de los bulbos también se ve deteriorada por las enfermedades. Sin embargo, este cultivo es apetecido por su sabor, considerado superior al del ajo importado de China. La pungencia del ajo está dada en parte por la acción de la enzima allinasa. Además, la resistencia a ciertos hongos patógenos está influenciada por la actividad de la enzima quitinasa. En el presente estudio se analizaron los genes que codifican para ambas enzimas, utilizando plántulas in vitro obtenidas a partir de materiales de las zonas de Llano Grande, Santa Ana, Miramar, San Ramón y de ajo importado de China. Se compararon y estudiaron las secuencias de ADN utilizando estos genes, con el fin de encontrar diferencias que permitieran la caracterización de distintos materiales. Los resultados obtenidos indicaron la presencia de distintas copias del gen allinasa. El gen de la quitinasa presentó una secuencia muy conservada en todos los materiales analizados. Se encontraron dos intrones altamente conservados en el germoplasma costarricense y el material de referencia asiático. Se concluyó que el ajo costarricense es muy similar al asiático. Y se presenta el primer informe de la existencia de intrones en la quitinasa del ajo.Garlic production in Costa Rica has been affected by the quality and quantity of the harvested seeds. Bulb production has also been deteriorated by diseases. However, this crop is preferred for its flavor, considered superior to the one imported from China. Pungency of garlic is partially due to the action of the alliinase enzyme. Furthermore, the resistance to certain pathogenic fungi is influenced by the chitinase enzyme activity. The encoding genes for both enzymes were analyzed in this study, by using in vitro plantlets obtained from local materials from Llano Grande, Santa Ana, Miramar and San Ramon zones and garlic imported

  15. Effects of Laser Peripheral Iridotomy in Subgroups of Primary Angle Closure Based on Iris Insertion

    OpenAIRE

    Sung-Cheol Yun; Ji Wook Hong; Kyung Rim Sung; Jin Young Lee

    2015-01-01

    Purpose. To investigate the effect of laser peripheral iridotomy (LPI) in subgroups of primary angle closure based on iris insertion configuration. Methods. Anterior segment optical coherence tomography (AS-OCT) images were obtained before and two weeks after LPI. Qualitative classification of angle closure eyes according to iris insertion (basal insertion group (BG) and nonbasal insertion group (NBG)) was performed. Anterior chamber depth (ACD), lens vault (LV), iris curvature, iris area, ir...

  16. Shotgun Cloning of Transposon Insertions in the Genome of Caenorhabditis elegans

    OpenAIRE

    Plasterk, Ronald H.A.; van der Linden, Alexander M.

    2006-01-01

    We present a strategy to identify and map large numbers of transposon insertions in the genome of Caenorhabditis elegans. Our approach makes use of the mutator strain mut-7, which has germline-transposition activity of the Tc1/mariner family of transposons, a display protocol to detect new transposon insertions, and the availability of the genomic sequence of C. elegans. From a pilot insertional mutagenesis screen, we have obtained 351 new Tc1 transposons inserted in or near 219 predicted C. ...

  17. Fc receptor endocytosis is controlled by a cytoplasmic domain determinant that actively prevents coated pit localization

    OpenAIRE

    1992-01-01

    Macrophages and B-lymphocytes express two major isoforms of Fc receptor (FcRII-B2 and FcRII-B1) that exhibit distinct capacities for endocytosis. This difference in function reflects the presence of an in- frame insertion of 47 amino acids in the cytoplasmic domain of the lymphocyte isoform (FcRII-B1) due to alternative mRNA splicing. By expressing wild type and mutant FcRII cDNAs in fibroblasts, we have now examined the mechanism by which the insertion acts to prevent coated pit localization...

  18. RHIC INSERTION REGION, SHUNT POWER SUPPLY CURRENT ERRORS

    International Nuclear Information System (INIS)

    The Relativistic Heavy Ion Collider (RHIC) was commissioned in 1999 and 2000. RHIC requires power supplies to supply currents to highly inductive superconducting magnets. The RHIC Insertion Region contain's many shunt power supplies to trim the current of different magnet elements in a large superconducting magnet circuit. Power Supply current error measurements were performed during the commissioning of RHIC. Models of these power supply systems were produced to predict and improve these power supply current errors using the circuit analysis program MicroCap V by Spectrum Software (TM). Results of the power supply current errors are presented from the models and from the measurements performed during the commissioning of RHIC

  19. Online robust model estimation during in vivo needle insertions.

    Science.gov (United States)

    Barbé, Laurent; Bayle, Bernard; de Mathelin, Michel

    2006-01-01

    Soft tissue modeling is of key importance in medical robotics and simulation. In the case of percutaneous operations, a fine model of layers transitions and target tissues is required. However, the nature and the variety of these tissues is such that this problem is extremely complex. In this article, we propose a method to estimate the interaction between in vivo tissues and a surgical needle. The online robust estimation of a varying parameters model is achieved during an insertion in standard operating conditions. PMID:16404010

  20. Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants

    Directory of Open Access Journals (Sweden)

    de Montaigu Amaury

    2011-07-01

    Full Text Available Abstract A method was developed to identify insertional mutants of Chlamydomonas reinhardtii disrupted for selected target genes. The approach relies on the generation of thousands of transformants followed by PCR-based screenings that allow for identification of strains harboring the introduced marker gene within specific genes of interest. Our results highlight the strengths and limitations of two independent screens that differed in the nature of the marker DNA used (PCR-amplified fragment containing the plasmid-free marker versus entire linearized plasmid with the marker and in the strategies used to maintain and store transformants.