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Sample records for cut family cdna

  1. Pattern analysis approach reveals restriction enzyme cutting abnormalities and other cDNA library construction artifacts using raw EST data

    Directory of Open Access Journals (Sweden)

    Zhou Sun

    2012-05-01

    Full Text Available Abstract Background Expressed Sequence Tag (EST sequences are widely used in applications such as genome annotation, gene discovery and gene expression studies. However, some of GenBank dbEST sequences have proven to be “unclean”. Identification of cDNA termini/ends and their structures in raw ESTs not only facilitates data quality control and accurate delineation of transcription ends, but also furthers our understanding of the potential sources of data abnormalities/errors present in the wet-lab procedures for cDNA library construction. Results After analyzing a total of 309,976 raw Pinus taeda ESTs, we uncovered many distinct variations of cDNA termini, some of which prove to be good indicators of wet-lab artifacts, and characterized each raw EST by its cDNA terminus structure patterns. In contrast to the expected patterns, many ESTs displayed complex and/or abnormal patterns that represent potential wet-lab errors such as: a failure of one or both of the restriction enzymes to cut the plasmid vector; a failure of the restriction enzymes to cut the vector at the correct positions; the insertion of two cDNA inserts into a single vector; the insertion of multiple and/or concatenated adapters/linkers; the presence of 3′-end terminal structures in designated 5′-end sequences or vice versa; and so on. With a close examination of these artifacts, many problematic ESTs that have been deposited into public databases by conventional bioinformatics pipelines or tools could be cleaned or filtered by our methodology. We developed a software tool for Abnormality Filtering and Sequence Trimming for ESTs (AFST, http://code.google.com/p/afst/ using a pattern analysis approach. To compare AFST with other pipelines that submitted ESTs into dbEST, we reprocessed 230,783 Pinus taeda and 38,709 Arachis hypogaea GenBank ESTs. We found 7.4% of Pinus taeda and 29.2% of Arachis hypogaea GenBank ESTs are “unclean” or abnormal, all of which could be cleaned

  2. Characterization of cDNA encoding human placental anticoagulant protein (PP4): Homology with the lipocortin family

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    Grundmann, U.; Abel, K.J.; Bohn, H.; Loebermann, H.; Lottspeich, F.; Kuepper, H. (Research Institutes, Postfach (West Germany))

    1988-06-01

    A cDNA library prepared from human placenta was screened for sequences encoding the placental protein 4 (PP4). PP4 is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade. Partial amino acid sequence information from PP4-derived cyanogen bromide fragments was used to design three oligonucleotide probes for screening the library. From 10{sup 6} independent recombinants, 18 clones were identified that hybridized to all three probes. These 18 recombinants contained cDNA inserts encoding a protein of 320 amino acid residues. In addition to the PP4 cDNA the authors identified 9 other recombinants encoding a protein with considerable similarity (74%) to PP4, which was termed PP4-X. PP4 and PP4-X belong to the lipocortin family, as judged by their homology to lipocortin I and calpactin I.

  3. Isolation, characterization and cDNA sequencing of a Kazal family proteinase inhibitor from seminal plasma of turkey (Meleagris gallopavo).

    Science.gov (United States)

    Słowińska, Mariola; Olczak, Mariusz; Wojtczak, Mariola; Glogowski, Jan; Jankowski, Jan; Watorek, Wiesław; Amarowicz, Ryszard; Ciereszko, Andrzej

    2008-06-01

    The turkey reproductive tract and seminal plasma contain a serine proteinase inhibitor that seems to be unique for the reproductive tract. Our experimental objective was to isolate, characterize and cDNA sequence the Kazal family proteinase inhibitor from turkey seminal plasma and testis. Seminal plasma contains two forms of a Kazal family inhibitor: virgin (Ia) represented by an inhibitor of moderate electrophoretic migration rate (present also in the testis) and modified (Ib, a split peptide bond) represented by an inhibitor with a fast migration rate. The inhibitor from the seminal plasma was purified by affinity, ion-exchange and reverse phase chromatography. The testis inhibitor was purified by affinity and ion-exchange chromatography. N-terminal Edman sequencing of the two seminal plasma inhibitors and testis inhibitor were identical. This sequence was used to construct primers and obtain a cDNA sequence from the testis. Analysis of a cDNA sequence indicated that turkey proteinase inhibitor belongs to Kazal family inhibitors (pancreatic secretory trypsin inhibitors, mammalian acrosin inhibitors) and caltrin. The turkey seminal plasma Kazal inhibitor belongs to low molecular mass inhibitors and is characterized by a high value of the equilibrium association constant for inhibitor/trypsin complexes.

  4. Characterization of cDNA from the miracidial antigen family of Schistosoma japonicum (Chinese strain)

    Institute of Scientific and Technical Information of China (English)

    余传信; 平山謙二; 朱荫昌; 菊池三惠子; 殷旭仁

    2003-01-01

    Objective To identify the egg antigens related to the formation of hepatic granulomas and fibrosis of Schistosomiasis japonica.Methods The egg cDNA library of Schistosoma japonicum (S.japonicum) was constructed and screened by immunological methods with the pooled sera of advanced schistosomiasis patients. The inserted foreign DNA fragments of positive clones were sequenced. The sequence data were analyzed using Wdnasis 2.5 and compared with Genebank data using blast software. Conclusion The cDNA sequence of the miracidial antigen of S.japonicum (Chinese strain) was obtained for the first time.

  5. Cutting

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    ... a traumatic experience, such as living through abuse , violence, or a disaster. Self-injury may feel like ... embarrassed." Sometimes self-injury affects a person's body image. Jen says, "I actually liked how the cuts ...

  6. cDNA cloning and functional characterization of ETHYLENE INSENSITIVE 3 orthologs from Oncidium Gower Ramsey involved in flower cutting and pollinia cap dislodgement.

    Science.gov (United States)

    Chen, Shin-Yu; Tsai, Hsing-Chun; Raghu, Rajasekaran; Do, Yi-Yin; Huang, Pung-Ling

    2011-10-01

    The cDNAs encoding ETHYLENE INSENSITIVE3 (EIN3) transcription factor, OgEIL1 and OgEIL2 of Oncidium were cloned, sequenced and characterized. The deduced amino acid sequences of OgEIL1 and OgEIL2 of identified cDNA clones contain all structural features found in the Arabidopsis EIN3, such as an amino terminal acidic domain, a proline-rich region, and five basic conserved domains. Complementation test for OgEIL1 in Arabidopsis ein3 mutant indicate that function of OgEIL1 is the same as Arabidopsis EIN3. RNA gel blot analysis indicated that OgEIL1 and OgEIL2 expressed differentially in the roots, stem, leaves and flower buds of Oncidium. OgEIL1 and OgEIL2 mRNA levels in fully opened flowers increased as time progressed after cutting and reached a maximum in the fifth day and decreased on seventh day, which is consistent with the hypothesis that flowers initiated to wilt when ethylene raised abruptly. In de-capped flowers, OgEIL2 mRNA showed a decrease, while OgEIL1 mRNA exhibited an increase. Exogenous application of ethylene increased the mRNA levels of OgEIL1 and OgEIL2 in flower buds and flowers after cutting compared prior to ethylene treatment, however, in pollinia de-capped flowers, both OgEIL1 and OgEIL2 mRNA levels responded to a decline to exogenous ethylene immediately after treatment. Collectively, it is suggested that the main functions of OgEIL1 and OgEIL2 are to modulate the senescence of Oncidium flowers.

  7. Molecular Cloning of a Novel cDNA From Mus Muscular BALB/c Mice Encoding Glycosyl Hydrolase Family 1: A Homolog of HumanLactase-Phlorizin Hydrolase

    Institute of Scientific and Technical Information of China (English)

    WEI HE; ZHEN-YU JI; CHENG-YU HUANG

    2006-01-01

    Objective To study the mechanism of lactose intolerance (LI) by cloning the mouse lactase cDNA and recombining a vector. Methods Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse (♂).Gene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase (LPH) in human, rat, and rabbit. A coding sequence (CDS) fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics. Results The cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinformatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosy1 hydrolase family 1 motif important for regulating lactase intolerance. Conclusion BALB/c mouse LPH cDNA (GenBank accession No: AY751548) provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.

  8. Structure and characterization of a cDNA clone for phenylalanine ammonia-lyase from cut-injured roots of sweet potato

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    Tanaka, Yoshiyuki; Matsuoka, Makoto; Yamanoto, Naoki; Ohashi, Yuko; Kano-Murakami, Yuriko; Ozeki, Yoshihiro (National Institute of Agro-Environmental Sciences, Ibaraki (Japan) Univ. of Tokyo (Japan))

    1989-08-01

    A cDNA clone for phenylalanine ammonia-lyase (PAL) induced in wounded sweet potato (Ipomoea batatas Lam.) root was obtained by immunoscreening a cDNA library. The protein produced in Escherichia coli cells containing the plasmid pPAL02 was indistinguishable from sweet potato PAL as judged by Ouchterlony double diffusion assays. The M{sub r} of its subunit was 77,000. The cells converted ({sup 14}C)-L-phenylalanine into ({sup 14}C)-t-cinnamic acid and PAL activity was detected in the homogenate of the cells. The activity was dependent on the presence of the pPAL02 plasmid DNA. The nucleotide sequence of the cDNA contained a 2,121-base pair (bp) open-reading frame capable of coding for a polypeptide with 707 amino acids (M{sub r} 77,137), a 22-bp 5{prime}-noncoding region and a 207-bp 3{prime}-noncoding region. The results suggest that the insert DNA fully encoded the amino acid sequence for sweet potato PAL that is induced by wounding. Comparison of the deduced amino acid sequence with that of a PAL cDNA fragment from Phaseolus vulgaris revealed 78.9% homology. The sequence from amino acid residues 258 to 494 was highly conserved, showing 90.7% homology.

  9. Cloning and expression of the cDNA encoding the FXPRL family of peptides and a functional analysis of their effect on breaking pupal diapause in Helicoverpa armigera.

    Science.gov (United States)

    Zhang, Tian-Yi; Sun, Jiu-Song; Zhang, Liu-Bin; Shen, Jin-Liang; Xu, Wei-Hua

    2004-01-01

    Diapause hormone (DH) and pheromone biosynthesis activating neuropeptide (PBAN) are encoded by a single mRNA in the suboesophegeal ganglion (SG) and are responsible for induction of embryonic diapause in Bombyx mori and sex pheromone biosynthesis in lepidopteran insects. PBAN cDNA analyses revealed that the DH-like peptide is present in several species that have a pupal diapause. However, the function of the DH-like peptide remains unknown. In the present study, we cloned the cDNA encoding DH-PBAN in Helicoverpa armigera utilizing the rapid amplification of the cDNA ends method. The nucleotide se quence analysis revealed that the longest open reading frame of this cDNA encodes a 194-amino acid precursor protein that con tains a 33-aa PBAN, a 24-aa DH-like peptide, and three other neuropeptides, all of which have a common C-terminal pentapeptide motif FXPR/KL ( X=G, T, S). A homology search showed that H. armigera DH-like and PBAN are highly homologous to those from other insects. Northern blot analysis demonstrated a single message RNA corresponding to the size of Har-DH-PBAN cDNA from pupal SG with significantly higher expression in the SG of nondiapause pupae than diapausing pupae. Western blot analysis showed DH-like peptide expression from SG of both males and females. When DH-like peptide was injected into nondiapause larvae and pupae, it did not induce diapause, but rather efficiently broke pupal diapause in H. armigera. The ED(50) of DH to terminate pupal diapause is 20 pmol/pupae. The other four FXPRLamide neuropeptides from the DH-PBAN polyprotein precursor have cross activity for diapause termination. These observations therefore suggest a potential role for these FXPRL family peptides in promoting continuous development in several noctuid species. The high expression of this gene in pharate adults and adults indicates that the FXPRL family peptides may have multiple physiological functions.

  10. A Müntz type theorem for a family of corner cutting schemes

    KAUST Repository

    Ait-Haddou, Rachid

    2013-02-01

    Dimension elevation process of Gelfond–Bézier curves generates a family of control polygons obtained through a sequence of corner cuttings. We give a Müntz type condition for the convergence of the generated control polygons to the underlying curve. The surprising emergence of the Müntz condition in the problem raises the question of a possible connection between the density questions of nested Chebyshev spaces and the convergence of the corresponding dimension elevation algorithms.

  11. Isolation and characterization of a cDNA clone encoding an auxin influx carrier in carnation cuttings. Expression in different organs and cultivars and its relationship with cold storage.

    Science.gov (United States)

    Oliveros-Valenzuela, María Del Rocío; Reyes, David; Sánchez-Bravo, José; Acosta, Manuel; Nicolás, Carlos

    2008-12-01

    Polar auxin transport (PAT) is necessary for the formation of adventitious roots in the base of leafy stem cuttings, as has been demonstrated in several studies in which the application of PAT inhibitors strongly inhibited the rooting of cuttings. However, unlike in the case of lateral roots, there is almost no information on the molecular mechanism that controls PAT in the formation of adventitious roots. A novel cDNA encoding an auxin influx carrier has been isolated and characterized from carnation (Dianthus caryophyllus) cuttings. The full length of DcAUX1 was obtained and the deduced aminoacid sequence revealed a high degree of identity with the corresponding auxin carrier proteins from several species. The expression of this gene depended on the organ, the carnation cultivar and the length of time cuttings had been stored in a cold chamber. As a rule, expression was higher in stem than in leaves, in the basal than in the first internode and in mature than in young leaves irrespective of the cultivar and the duration of the storage. This pattern of expression agrees with the results of a previous study showing that auxin from mature leaves was essential for rooting, while exogenous auxin applied to mature leaves was polarly transported in the stem and accumulated in the basal internode (the rooting zone). Variations in the expression observed during storage (depending of the cultivar) might be related to the variation in PAT and rooting reported in previous studies.

  12. Cloning of a cDNA encoding a novel human nuclear phosphoprotein belonging to the WD-40 family

    DEFF Research Database (Denmark)

    Honoré, B; Leffers, H; Madsen, Peder

    1994-01-01

    We have cloned and expressed in vaccinia virus a cDNA encoding an ubiquitous 501-amino-acid (aa) phosphoprotein that corresponds to protein IEF SSP 9502 (79,400 Da, pI 4.5) in the master 2-D-gel keratinocyte protein database [Celis et al., Electrophoresis 14 (1993) 1091-1198]. The deduced aa...

  13. A cDNA cloned from Physarum polycephalum encodes new type of family 3 beta-glucosidase that is a fusion protein containing a calx-beta motif.

    Science.gov (United States)

    Maekawa, Akinori; Hayase, Masato; Yubisui, Toshitsugu; Minami, Yoshiko

    2006-01-01

    The microplasmodia of Physarum polycephalum express three types of beta-glucosidases: secretory enzyme, a soluble cytoplasmic enzyme and a membrane-bound enzyme. We are interested in the physiological role of three enzymes. We report the sequence of cDNA for membrane beta-glucosidase 1, which consists of 3825 nucleotides that includes an open reading frame encoding 1248 amino acids. The molecular weight of membrane beta-glucosidase 1 was calculated to be 131,843 based on the predicted amino acid composition. Glycosyl hydrolase family 3 N-terminal and C-terminal domains were found within the N-terminal half of the membrane beta-glucosidase 1 sequence and were highly homologous with the primary structures of fungal beta-glucosidases. Notably, the C-terminal half of membrane beta-glucosidase 1 contains two calx-beta motifs, which are known to be Ca(2+) binding domains in the Drosophila Na(+)/Ca(2+) exchanger; an RGD sequence, which is known to be a cell attachment sequence; and a transmembrane region. In this way, Physarum membrane beta-glucosidase 1 differs from all previously identified family 3 beta-glucosidases. In addition to cDNA for membrane beta-glucosidase 1, two other distinctly different mRNAs were also isolated. Two sequences were largely identical to cDNA for membrane beta-glucosidase 1, but included a long insert sequence having a stop codon, leading to truncation of their products, which could account for other beta-glucosidase forms occurred in Physarum poycephalum. Thus, the membrane beta-glucosidase is a new type family 3 enzyme fused with the Calx-beta domain. We propose that Calx-beta domain may modulate the beta-glucosidase activity in response to changes in the Ca(2+) concentration.

  14. hSmad5 gene, a human hSmad family member: its full length cDNA, genomic structure, promoter region and mutation analysis in human tumors.

    Science.gov (United States)

    Gemma, A; Hagiwara, K; Vincent, F; Ke, Y; Hancock, A R; Nagashima, M; Bennett, W P; Harris, C C

    1998-02-19

    hSmad (mothers against decapentaplegic)-related proteins are important messengers within the Transforming Growth Factor-beta1 (TGF-beta1) superfamily signal transduction pathways. To further characterize a member of this family, we obtained a full length cDNA of the human hSmad5 (hSmad5) gene by rapid amplification of cDNA ends (RACE) and then determined the genomic structure of the gene. There are eight exons and two alternative transcripts; the shorter transcript lacks exon 2. We identified the hSmad5 promoter region from a human genomic YAC clone by obtaining the nucleotide sequence extending 1235 base pairs upstream of the 5' end of the cDNA. We found a CpG island consistent with a promoter region, and we demonstrated promoter activity in a 1232 bp fragment located upstream of the transcription initiation site. To investigate the frequency of somatic hSmad5 mutations in human cancers, we designed intron-based primers to examine coding regions by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. Neither homozygous deletions or point mutations were found in 40 primary gastric tumors and 51 cell lines derived from diverse types of human cancer including 20 cell lines resistant to the growth inhibitory effects of TGF-beta1. These results suggest that the hSmad5 gene is not commonly mutated and that other genetic alterations mediate the loss of TGF-beta1 responsiveness in human cancers.

  15. Avocado cellulase: nucleotide sequence of a putative full-length cDNA clone and evidence for a small gene family.

    Science.gov (United States)

    Tucker, M L; Durbin, M L; Clegg, M T; Lewis, L N

    1987-05-01

    A cDNA library was prepared from ripe avocado fruit (Persea americana Mill. cv. Hass) and screened for clones hybridizing to a 600 bp cDNA clone (pAV5) coding for avocado fruit cellulase. This screening led to the isolation of a clone (pAV363) containing a 2021 nucleotide transcribed sequence and an approximately 150 nucleotide poly(A) tail. Hybridization of pAV363 to a northern blot shows that the length of the homologous message is approximately 2.2 kb. The nucleotide sequence of this putative full-length mRNA clone contains an open reading frame of 1482 nucleotides which codes for a polypeptide of 54.1 kD. The deduced amino acid composition compares favorably with the amino acid composition of native avocado cellulase determined by amino acid analysis. Southern blot analysis of Hind III and Eco RI endonuclease digested genomic DNA indicates a small family of cellulase genes.

  16. Isolation of a cDNA encoding thymic shared antigen-1. A new member of the Ly6 family with a possible role in T cell development.

    Science.gov (United States)

    MacNeil, I; Kennedy, J; Godfrey, D I; Jenkins, N A; Masciantonio, M; Mineo, C; Gilbert, D J; Copeland, N G; Boyd, R L; Zlotnik, A

    1993-12-15

    We have previously characterized a novel mouse thymocyte marker, defined as thymic shared Ag-1 (TSA-1), present on both immature thymocytes and a subset of thymic medullary epithelial cells. MTS 35, a mAb specific for TSA-1, alters T cell differentiation when added to fetal thymic organ cultures, suggesting TSA-1 may be important for T cell development in the thymus. In this study, we describe the isolation of a cDNA encoding TSA-1 using transient expression of COS-7 cells and selection with MTS 35. The predicted amino acid sequence of this cDNA encodes a 15 to 17-kDa protein and the expressed protein is linked to the membrane via a phosphatidylinositol moiety. TSA-1 is transcriptionally active at various levels in all organs examined, suggesting that its role is not solely intrathymic. TSA-1 shares amino acid sequence homology to the mouse Ly6 multigene family, epidermal growth factor-like receptors, and to cobra venom neurotoxin. The Tsa-1 locus is located on chromosome 15 linked to Ly6 on the mouse genome. We also examined the effects of MTS 35 in fetal thymic organ cultures repopulated with two subsets of thymocytes representing defined stages of T cell development. Our results suggest that TSA-1 may play a role during positive selection and the transition from CD4+CD8+ thymocytes to the mature CD4+CD8- and CD4-CD8+ subsets.

  17. Expression of NGF family and their receptors in gastric carcinoma:A cDNA microarray study

    Institute of Scientific and Technical Information of China (English)

    Jian-Jun Du; Ying-Bin Liu; Ze-Guang Han; Ke-Feng Dou; Shu-You Peng; Bing-Zhi Qian; Hua-Sheng Xiao; Feng Liu; Wei-Zhong Wang; Wen-Xian Guan; Zhi-Qing Gao

    2003-01-01

    AIM: To investigate the expression of NGF family and their receptors in gastric carcinoma and normal gastric mucosa,and to elucidate their effects on gastric carcinoma.METHODS: RNA of gastric cancer tissues and normal gastric tissues was respectively isolated and mRNA was purified.Probes of both mRNA reverse transcription product cDNAs labled with α-33P dATP were respectively hybridized with Atlas Array membrane where NGF and their family genes were spotted on. Hybridized signal images were scanned on phosphor screen with ImageQuant 5.1 software after hybridization. Normalized values on spots were analyzed with ArrayVersion 5.0 software. Differential expression of NGF family and their receptors mRNA was confirmed between hybridized Atlas Array membranes of gastric cancer tissues and normal gastric mucosa, then their effects on gastric carcinoma were investigated.RESULTS: Hybridization signal images on Atlas Array membrane appeared in a lower level of nonspecific hybridization. Both of NGF family and their receptors Trk family mRNA were expressed in gastric cancer and normal gastric mucosa. But adversely up-regulated expression in other tissues and organs. NGF, BDGF, NT-3, NT-4/5, NT-6and TrkA, B and C were down-regulated simultaneously in gastric carcinoma in comparison with normal gastric mucosa. Degrees of down-regulation in NGF family were greater than those in their receptors Trk family. Downregulation of NT-3 and BDGF was the most significant,and TrkC down-regulation level was the lowest in receptors Trk family.CONCLUSION: Down-regulated expression of NGF family and their receptors Trk family mRNA in gastric cancer is confirmed. NGF family and their receptors Trk family probably play a unique role in gastric cancer cell apoptosis by a novel Ras or Raf signal transduction pathway. Their synchronous effects are closely associated with occurrence and development of gastric carcinoma induced by reduction of signal transduction of programmed cell death.

  18. Amino acid sequence of Coprinus macrorhizus peroxidase and cDNA sequence encoding Coprinus cinereus peroxidase. A new family of fungal peroxidases.

    Science.gov (United States)

    Baunsgaard, L; Dalbøge, H; Houen, G; Rasmussen, E M; Welinder, K G

    1993-04-01

    Sequence analysis and cDNA cloning of Coprinus peroxidase (CIP) were undertaken to expand the understanding of the relationships of structure, function and molecular genetics of the secretory heme peroxidases from fungi and plants. Amino acid sequencing of Coprinus macrorhizus peroxidase, and cDNA sequencing of Coprinus cinereus peroxidase showed that the mature proteins are identical in amino acid sequence, 343 residues in size and preceded by a 20-residue signal peptide. Their likely identity to peroxidase from Arthromyces ramosus is discussed. CIP has an 8-residue, glycine-rich N-terminal extension blocked with a pyroglutamate residue which is absent in other fungal peroxidases. The presence of pyroglutamate, formed by cyclization of glutamine, and the finding of a minor fraction of a variant form lacking the N-terminal residue, indicate that signal peptidase cleavage is followed by further enzymic processing. CIP is 40-45% identical in amino-acid sequence to 11 lignin peroxidases from four fungal species, and 42-43% identical to the two known Mn-peroxidases. Like these white-rot fungal peroxidases, CIP has an additional segment of approximately 40 residues at the C-terminus which is absent in plant peroxidases. Although CIP is much more similar to horseradish peroxidase (HRP C) in substrate specificity, specific activity and pH optimum than to white-rot fungal peroxidases, the sequences of CIP and HRP C showed only 18% identity. Hence, CIP qualifies as the first member of a new family of fungal peroxidases. The nine invariant residues present in all plant, fungal and bacterial heme peroxidases are also found in CIP. The present data support the hypothesis that only one chromosomal CIP gene exists. In contrast, a large number of secretory plant and fungal peroxidases are expressed from several peroxidase gene clusters. Analyses of three batches of CIP protein and of 49 CIP clones revealed the existence of only two highly similar alleles indicating less

  19. Cloning and molecular characterization of cDNA encoding a mouse male-enhanced antigen-2 (Mea-2): a putative family of the Golgi autoantigen.

    Science.gov (United States)

    Kondo, M; Sutou, S

    1997-01-01

    The male-enhanced antigen-2 (Mea-2) gene was originally identified with a monoclonal histocompatibility Y (H-Y) antibody (mAb4VII). There is no report of the full length cDNA encode for Mea-2 product until this report. In this study, we isolated the full length mouse Mea-2 cDNA by screening a testis cDNA library with a PCR-amplified Mea-2 product, and direct PCR amplification of its upstream sequences from the cDNA library. The primary structure of the Mea-2 peptide, deduced from this nucleotide sequence, shows that it encode a 150 kDa protein, of 1325 amino acid residues, which contained five putative N-glycosylation sites and four leucine zipper motifs. A data bank search indicated that it has high homology with a human Golgi autoantigen (golgin-160) both in its nucleotides (78%) and amino acids sequence (83%). This suggests that Mea-2 gene product may encode a golgi structural protein. In situ hybridization analysis suggested that the Mea-2 gene is expressed in spermatids during spermatogenesis as already shown by Mea-1, suggesting that Mea-2 gene product as well as Mea-1 have also some role for spermatogenesis.

  20. Molecular Descriptors Family on Vertex Cutting: Relationships between Acelazolamide Structures and their Inhibitory Activity

    Directory of Open Access Journals (Sweden)

    Sorana D. BOLBOACĂ

    2009-12-01

    Full Text Available Aim: To investigate the relationship between the structural information of acetazolamides and their inhibitory activity on carbonic anhydrase II. Material and Method: A sample of previously reported acetazolamides was studied. A pool of descriptors was calculated based on matrix representation and vertex cut in order to be included in the multiple linear regression analysis. The best performing model in terms of goodness-of-fit was analysed in order to assess its validity and reliability. The model was compared with previously reported models using a series of information and prediction criteria besides the Steiger’s Z test. Results: A model with a 99.77% determination coefficient proved to be the best performing model. The obtained model proved to have a less than 5% average of the absolute difference between the observed and the estimated inhibitory activity. The information and prediction criteria showed that the obtained model was better than the previously reported models. This conclusion is also sustained by the results of Steiger’s Z test (7.78; p = 3.66·10-15. Conclusion: The inhibitory activity on carbonic anhydrase II of acetazolamides proved to be of geometric and topologic nature and depended on the compounds’ melting point, relative atomic mass and atomic electronegativity.

  1. Sequencing analysis of 20,000 full-length cDNA clones from cassava reveals lineage specific expansions in gene families related to stress response

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    Sakaki Yoshiyuki

    2007-12-01

    Full Text Available Abstract Background Cassava, an allotetraploid known for its remarkable tolerance to abiotic stresses is an important source of energy for humans and animals and a raw material for many industrial processes. A full-length cDNA library of cassava plants under normal, heat, drought, aluminum and post harvest physiological deterioration conditions was built; 19968 clones were sequence-characterized using expressed sequence tags (ESTs. Results The ESTs were assembled into 6355 contigs and 9026 singletons that were further grouped into 10577 scaffolds; we found 4621 new cassava sequences and 1521 sequences with no significant similarity to plant protein databases. Transcripts of 7796 distinct genes were captured and we were able to assign a functional classification to 78% of them while finding more than half of the enzymes annotated in metabolic pathways in Arabidopsis. The annotation of sequences that were not paired to transcripts of other species included many stress-related functional categories showing that our library is enriched with stress-induced genes. Finally, we detected 230 putative gene duplications that include key enzymes in reactive oxygen species signaling pathways and could play a role in cassava stress response features. Conclusion The cassava full-length cDNA library here presented contains transcripts of genes involved in stress response as well as genes important for different areas of cassava research. This library will be an important resource for gene discovery, characterization and cloning; in the near future it will aid the annotation of the cassava genome.

  2. Infectious Maize rayado fino virus from cloned cDNA

    Science.gov (United States)

    Maize rayado fino virus (MRFV) is the type member of the marafiviruses within the family Tymoviridae. A cDNA clone from which infectious RNA can be transcribed was produced from a US isolate of MRFV (MRFV-US). Infectivity of transcripts derived from cDNA clones was demonstrated by infection of mai...

  3. Four rice seed cDNA clones belonging to the alpha-amylase/trypsin inhibitor gene family encode potential rice allergens.

    Science.gov (United States)

    Alvarez, A M; Fukuhara, E; Nakase, M; Adachi, T; Aoki, N; Nakamura, R; Matsuda, T

    1995-07-01

    Four rice seed proteins encoded by cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family were overexpressed as TrpE-fusion proteins in E. coli. The expressed rice proteins were detected by SDS-PAGE as major proteins in bacterial cell lysates. Western blot analyses showed that all the recombinant proteins were immunologically reactive to rabbit polyclonal antibodies and to a mouse monoclonal antibody (25B9) specific for a previously isolated rice allergen of 16 kDa. Some truncated proteins from deletion mutants of the cDNAs retained their reactivity to the specific antibodies. These results suggest that the cDNAs encode potential rice allergens and that some epitopes of the recombinant proteins are still immunoreactive when they are expressed as their fragments.

  4. Nucleotide sequence of a cDNA clone encoding a major allergenic protein in rice seeds. Homology of the deduced amino acid sequence with members of alpha-amylase/trypsin inhibitor family.

    Science.gov (United States)

    Izumi, H; Adachi, T; Fujii, N; Matsuda, T; Nakamura, R; Tanaka, K; Urisu, A; Kurosawa, Y

    1992-05-18

    A cDNA clone of rice major allergenic protein (RAP) was isolated from a cDNA library of maturing rice seeds. The cDNA had an open reading frame (486 nucleotides) which coded a 162 amino acid residue polypeptide comprising a 27-residue signal peptide and a 135-residue mature protein of M(r) 14,764. The deduced amino acid sequence of RAP showed a considerable similarity to barley trypsin inhibitor [1983, J. Biol. Chem. 258, 7998-8003] and wheat alpha-amylase inhibitor [1981, Phytochemistry 20, 1781-1784].

  5. Normalized cDNA libraries

    Science.gov (United States)

    Soares, Marcelo B.; Efstratiadis, Argiris

    1997-01-01

    This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to moderate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library.

  6. Construction of cDNA representational difference analysis based on two cDNA libraries and identification of garlic inducible expression genes in human gastric cancer cells

    Institute of Scientific and Technical Information of China (English)

    Yong Li; Lin Yang; Jian-Tao Cui; Wen-Mei Li; Rui-Fang Guo; You-Yong Lu

    2002-01-01

    AIM: To elucidate molecular mechanism of chemopreventiveefficacies of garlic against human gastric cancer (HGC):METHODS: HGC cell line BGC823 was treated with Allitridi (akind of garlic extract) and Allitridi-treated and parentalBGC823 cDNA librarles were constructed respectively byusing λZAP Ⅱ vector. cDNA Representatinal DifferenceAnalysis (cDNA RDA) was perfonmed using BamH Ⅰ cutting-site and abundant ~DNA messages provided by the Iibrarles.Northern blot analysls was applied to identifythe obtaineddifference prnducts.RESULTS: Two specific cDNA fragments were obtained andcharacterized to be derived from homo sapiens folatereceptorα (FRα) gene and calcyclin gene respectively.Northern blot results showed a 4-fold increase in FRα geneexpression level and 9-fold increase in calcyclin mRNA levelin BGC823 cells after Allilridi treatment for 72 h.CONCLUSION: The method of cDNA RDA based on cDNAlibraries combines the high specificity of cDNA RDA withabundant cDNA messages in cDNA library; this expands theapplication of cDNA library and increases the specificity ofcDNA RDA. Up-regulstion of FRα gene and calcyclin geneexpressions induced by Allitridi provide valuable molecularevidence for theefficacy of garlic in treating HGC as well asother diseases.

  7. Cutting assembly

    Science.gov (United States)

    Racki, Daniel J.; Swenson, Clark E.; Bencloski, William A.; Wineman, Arthur L.

    1984-01-01

    A cutting apparatus includes a support table mounted for movement toward and away from a workpiece and carrying a mirror which directs a cutting laser beam onto the workpiece. A carrier is rotatably and pivotally mounted on the support table between the mirror and workpiece and supports a conduit discharging gas toward the point of impingement of the laser beam on the workpiece. Means are provided for rotating the carrier relative to the support table to place the gas discharging conduit in the proper positions for cuts made in different directions on the workpiece.

  8. Bone cutting.

    Science.gov (United States)

    Giraud, J Y; Villemin, S; Darmana, R; Cahuzac, J P; Autefage, A; Morucci, J P

    1991-02-01

    Bone cutting has always been a problem for surgeons because bone is a hard living material, and many osteotomes are still very crude tools. Technical improvement of these surgical tools has first been their motorization. Studies of the bone cutting process have indicated better features for conventional tools. Several non-conventional osteotomes, particularly ultrasonic osteotomes are described. Some studies on the possible use of lasers for bone cutting are also reported. Use of a pressurised water jet is also briefly examined. Despite their advantages, non-conventional tools still require improvement if they are to be used by surgeons.

  9. Cutting Cosmos

    DEFF Research Database (Denmark)

    Mikkelsen, Henrik Hvenegaard

    The foundation for this book is an ethnographic study of masculinity in a Bugkalot village in northern Philippines. While offering new research on the Bugkalot, widely known as the Ilongot, more than 30 years after the last important works were written on this famous hill-people, Cutting Cosmos...... into egalitarian relations. Cutting Cosmos shows how these seemingly opposed characteristics of male life - the egalitarianism and the assertive ideals - are interwoven. Acts of dominance are presented as acts of transgression that are persistently ritualized, contained and isolated as spectacular events within...

  10. Cloning and expression of a novel human HCUTA cDNA

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Copper is one of the most important trace elements to life. Human HCUTA is a novel cDNA encoding a 156aa protein, which may participate in human copper tolerance system. The HCUTA protein is highly similar to protein CUT A1 of E. coli. The whole opening reading frame of HCUTA cDNA was amplified by PCR and cloned into pET28a + express vector, and the HCUTA protein was effectively expressed in E. coli BL21 (DE3).

  11. cDNA sequence quality data - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project cDNA sequence quality data Data detail Data name cDNA sequence quality... data Description of data contents Phred's quality score. PHD format, one file to a single cDNA data, and co...ription Download License Update History of This Database Site Policy | Contact Us cDNA sequence quality data - Budding yeast cDNA sequencing project | LSDB Archive ...

  12. Biological characterization of liver fatty acid binding gene from miniature pig liver cDNA library.

    Science.gov (United States)

    Gao, Y H; Wang, K F; Zhang, S; Fan, Y N; Guan, W J; Ma, Y H

    2015-01-01

    Liver fatty acid binding proteins (L-FABP) are a family of small, highly conserved, cytoplasmic proteins that bind to long-chain fatty acids and other hydrophobic ligands. In this study, a full-length enriched cDNA library was successfully constructed from Wuzhishan miniature pig, and then the L-FABP gene was cloned from this cDNA library and an expression vector (pEGFP-N3-L-FABP) was constructed in vitro. This vector was transfected into hepatocytes to test its function. The results of western blotting analysis demonstrated that the L-FABP gene from our full-length enriched cDNA library regulated downstream genes, including the peroxisome proliferator-activated receptor family in hepatocytes. This study provides a theoretical basis and experimental evidence for the application of L-FABP for the treatment of liver injury.

  13. Multibeam fiber laser cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove; Hansen, Klaus Schütt; Nielsen, Jakob Skov

    2009-01-01

    The appearance of the high power high brilliance fiber laser has opened for new possibilities in laser materials processing. In laser cutting this laser has demonstrated high cutting performance compared to the dominating Cutting laser, the CO2 laser. However, quality problems in fiber-laser...... cutting have until now limited its application to metal cutting. In this paper the first results of proof-of-principle Studies applying a new approach (patent pending) for laser cutting with high brightness and short wavelength lasers will be presented. In the approach, multibeam patterns are applied...... to control the melt flow out of the cut kerf resulting in improved cut quality in metal cutting. The beam patterns in this study are created by splitting up beams from two single mode fiber lasers and combining these beams into a pattern in the cut kerf. The results are obtained with a total of 550 W...

  14. Multibeam Fibre Laser Cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    The appearance of the high power high brilliance fibre laser has opened for new possibilities in laser materials processing. In laser cutting this laser has demonstrated high cutting performance compared to the dominating cutting laser, the CO2-laser. However, quality problems in fibre-laser...... cutting have until now limited its application in metal cutting. In this paper the first results of proof-of-principle studies applying a new approach (patent pending) for laser cutting with high brightness short wavelength lasers will be presented. In the approach, multi beam patterns are applied...... to control the melt flow out of the cut kerf resulting in improved cut quality in metal cutting. The beam patterns in this study are created by splitting up beams from 2 single mode fibre lasers and combining these beams into a pattern in the cut kerf. The results are obtained with a total of 550 W of single...

  15. Molecular Cloning and Sequencing of Channel Catfish, Ictalurus punctatus, Cathepsin H and L cDNA

    Science.gov (United States)

    Cathepsin H and L, a lysosomal cysteine endopeptidase of the papain family, are ubiquitously expressed and involve in antigen processing. In this communication, the channel catfish cathepsin H and L transcripts were sequenced and analyzed. Total RNA from tissues was extracted and cDNA libraries we...

  16. Characterization of the soybean early nodulin cDNA clone GmENOD55.

    NARCIS (Netherlands)

    Blank, de C.; Mylona, P.; Katinakis, P.; Bisseling, T.; Franssen, H.; Yang, W.C.

    1993-01-01

    Two cDNA clones of the soybean early nodulin GmENOD55 were characterized. These clones may represent two members of the soybean early nodulin gene family GmENOD55. GmENOD55 has an N-terminal signal peptide and it contains an internal domain consisting of proline and serine residues. Analyses of

  17. Flexible Laser Metal Cutting

    DEFF Research Database (Denmark)

    Villumsen, Sigurd; Jørgensen, Steffen Nordahl; Kristiansen, Morten

    2014-01-01

    This paper describes a new flexible and fast approach to laser cutting called ROBOCUT. Combined with CAD/CAM technology, laser cutting of metal provides the flexibility to perform one-of-a-kind cutting and hereby realises mass production of customised products. Today’s laser cutting techniques...... possess, despite their wide use in industry, limitations regarding speed and geometry. Research trends point towards remote laser cutting techniques which can improve speed and geometrical freedom and hereby the competitiveness of laser cutting compared to fixed-tool-based cutting technology...... such as punching. This paper presents the concepts and preliminary test results of the ROBOCUT laser cutting technology, a technology which potentially can revolutionise laser cutting....

  18. Cutting state identification

    Energy Technology Data Exchange (ETDEWEB)

    Berger, B.S.; Minis, I.; Rokni, M. [Univ. of Maryland, College Park, MD (United States)] [and others

    1997-12-31

    Cutting states associated with the orthogonal cutting of stiff cylinders are identified through an analysis of the singular values of a Toeplitz matrix of third order cumulants of acceleration measurements. The ratio of the two pairs of largest singular values is shown to differentiate between light cutting, medium cutting, pre-chatter and chatter states. Sequences of cutting experiments were performed in which either depth of cut or turning frequency was varied. Two sequences of experiments with variable turning frequency and five with variable depth of cut, 42 cutting experiments in all, provided a database for the calculation of third order cumulants. Ratios of singular values of cumulant matrices find application in the analysis of control of orthogonal cutting.

  19. Cloning and expression of a cDNA encoding ribosomal protein S4 from Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A cDNA clone, pS4, has been isolated from a cDNA library prepared from rice anthers of about 1.0 mm in length. DNA sequence analysis and database search show that the cDNA encodes a protein which is highly homologous to eukaryotic 80S ribosomal protein subunit 4 (S4). Northern hybridization indicates that this gene expresses in all tissues analyzed although the expression level varies and it cannot be induced by mechanical wounding in leaves. Southern blot analysis demonstrates that this rice S4 gene is from a multigene family.

  20. Simulation of Laser Cutting

    Science.gov (United States)

    Schulz, Wolfgang; Nießen, Markus; Eppelt, Urs; Kowalick, Kerstin

    Laser cutting is a thermal separation process widely used in shaping and contour cutting applications. There are, however, gaps in understanding the dynamics of the process, especially issues related to cut quality. This work describes the advances in fundamental physical modelling and process monitoring of laser cutting, as well as time varying processes such as contour cutting. Diagnosis of ripple and dross formation is advanced to observe the melt flow and its separation simultaneously as well as the spatial shape of the cut kerf.

  1. Simple and inexpensive three-step rapid amplification of cDNA 5′ ends using 5′ phosphorylated primers

    OpenAIRE

    Dallmeier, Kai; Neyts, Johan

    2013-01-01

    Rapid amplification of cDNA 5′ ends (5′-RACE) is routinely used for the sequence analysis of the upstream noncoding regions of cellular mRNAs; however, it represents a tedious and cost-intensive procedure. By employing 5′ phosphorylated gene-specific primers for first-strand cDNA synthesis, we cut short the previously established reverse ligation and amplification protocol of Mandl and coworkers (BioTechniques, 1991, vol. 10, pp. 484–486) to a streamlined three-step procedure that no longer d...

  2. Cutting edge: Ly9 (CD229), a SLAM family receptor, negatively regulates the development of thymic innate memory-like CD8+ T and invariant NKT cells.

    Science.gov (United States)

    Sintes, Jordi; Cuenca, Marta; Romero, Xavier; Bastos, Ricardo; Terhorst, Cox; Angulo, Ana; Engel, Pablo

    2013-01-01

    Signaling lymphocytic activation molecule family receptors and the specific adapter signaling lymphocytic activation molecule-associated protein modulate the development of innate-like lymphocytes. In this study, we show that the thymus of Ly9-deficient mice contains an expanded population of CD8 single-positive cells with the characteristic phenotype of innate memory-like CD8(+) T cells. Moreover, the proportion of these innate CD8(+) T cells increased dramatically postinfection with mouse CMV. Gene expression profiling of Ly9-deficient mice thymi showed a significant upregulation of IL-4 and promyelocytic leukemia zinc finger. Analyses of Ly9(-/-)IL4ra(-/-) double-deficient mice revealed that IL-4 was needed to generate the thymic innate CD8(+) T cell subset. Furthermore, increased numbers of invariant NKT cells were detected in Ly9-deficient thymi. In wild-type mice, IL-4 levels induced by α-galactosylceramide injection could be inhibited by a mAb against Ly9. Thus, Ly9 plays a unique role as an inhibitory cell surface receptor regulating the size of the thymic innate CD8(+) T cell pool and the development of invariant NKT cells.

  3. Changing cultural attitudes towards female genital cutting.

    Science.gov (United States)

    Vogt, Sonja; Mohmmed Zaid, Nadia Ahmed; El Fadil Ahmed, Hilal; Fehr, Ernst; Efferson, Charles

    2016-10-27

    As globalization brings people with incompatible attitudes into contact, cultural conflicts inevitably arise. Little is known about how to mitigate conflict and about how the conflicts that occur can shape the cultural evolution of the groups involved. Female genital cutting is a prominent example. Governments and international agencies have promoted the abandonment of cutting for decades, but the practice remains widespread with associated health risks for millions of girls and women. In their efforts to end cutting, international agents have often adopted the view that cutting is locally pervasive and entrenched. This implies the need to introduce values and expectations from outside the local culture. Members of the target society may view such interventions as unwelcome intrusions, and campaigns promoting abandonment have sometimes led to backlash as they struggle to reconcile cultural tolerance with the conviction that cutting violates universal human rights. Cutting, however, is not necessarily locally pervasive and entrenched. We designed experiments on cultural change that exploited the existence of conflicting attitudes within cutting societies. We produced four entertaining movies that served as experimental treatments in two experiments in Sudan, and we developed an implicit association test to unobtrusively measure attitudes about cutting. The movies depart from the view that cutting is locally pervasive by dramatizing members of an extended family as they confront each other with divergent views about whether the family should continue cutting. The movies significantly improved attitudes towards girls who remain uncut, with one in particular having a relatively persistent effect. These results show that using entertainment to dramatize locally discordant views can provide a basis for applied cultural evolution without accentuating intercultural divisions.

  4. Device for cutting protrusions

    Science.gov (United States)

    Bzorgi, Fariborz M.

    2011-07-05

    An apparatus for clipping a protrusion of material is provided. The protrusion may, for example, be a bolt head, a nut, a rivet, a weld bead, or a temporary assembly alignment tab protruding from a substrate surface of assembled components. The apparatus typically includes a cleaver having a cleaving edge and a cutting blade having a cutting edge. Generally, a mounting structure configured to confine the cleaver and the cutting blade and permit a range of relative movement between the cleaving edge and the cutting edge is provided. Also typically included is a power device coupled to the cutting blade. The power device is configured to move the cutting edge toward the cleaving edge. In some embodiments the power device is activated by a momentary switch. A retraction device is also generally provided, where the retraction device is configured to move the cutting edge away from the cleaving edge.

  5. Lectin cDNA and transgenic plants derived therefrom

    Science.gov (United States)

    Raikhel, Natasha V.

    2000-10-03

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

  6. Graphs of Plural Cuts

    CERN Document Server

    Dosen, K

    2011-01-01

    Plural (or multiple-conclusion) cuts are inferences made by applying a structural rule introduced by Gentzen for his sequent formulation of classical logic. As singular (single-conclusion) cuts yield trees, which underlie ordinary natural deduction derivations, so plural cuts yield graphs of a more complicated kind, related to trees, which this paper defines. Besides the inductive definition of these oriented graphs, which is based on sequent systems, a non-inductive, graph-theoretical, combinatorial, definition is given, and to reach that other definition is the main goal of the paper. As trees underlie multicategories, so the graphs of plural cuts underlie polycategories. The graphs of plural cuts are interesting in particular when the plural cuts are appropriate for sequent systems without the structural rule of permutation, and the main body of the paper deals with that matter. It gives a combinatorial characterization of the planarity of the graphs involved.

  7. Laser Cutting, Development Trends

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    1999-01-01

    In this paper a short review of the development trends in laser cutting will be given.The technology, which is the fastest expanding industrial production technology will develop in both its core market segment: Flat bed cutting of sheet metal, as it will expand in heavy industry and in cutting o...... of 3-dimensional shapes.The CO2-laser will also in the near future be the dominating laser source in the market, although the new developments in ND-YAG-lasers opens for new possibilities for this laser type.......In this paper a short review of the development trends in laser cutting will be given.The technology, which is the fastest expanding industrial production technology will develop in both its core market segment: Flat bed cutting of sheet metal, as it will expand in heavy industry and in cutting...

  8. Laser cutting plastic materials

    Energy Technology Data Exchange (ETDEWEB)

    Van Cleave, R.A.

    1980-08-01

    A 1000-watt CO/sub 2/ laser has been demonstrated as a reliable production machine tool for cutting of plastics, high strength reinforced composites, and other nonmetals. More than 40 different plastics have been laser cut, and the results are tabulated. Applications for laser cutting described include fiberglass-reinforced laminates, Kevlar/epoxy composites, fiberglass-reinforced phenolics, nylon/epoxy laminates, ceramics, and disposable tooling made from acrylic.

  9. Ultrasonic Cutting of Foods

    Science.gov (United States)

    Schneider, Yvonne; Zahn, Susann; Rohm, Harald

    In the field of food engineering, cutting is usually classified as a mechanical unit operation dealing with size reduction by applying external forces on a bulk product. Ultrasonic cutting is realized by superpositioning the macroscopic feed motion of the cutting device or of the product with a microscopic vibration of the cutting tool. The excited tool interacts with the product and generates a number of effects. Primary energy concentration in the separation zone and the modification of contact friction along the tool flanks arise from the cyclic loading and are responsible for benefits such as reduced cutting force, smooth cut surface, and reduced product deformation. Secondary effects such as absorption and cavitation originate from the propagation of the sound field in the product and are closely related to chemical and physical properties of the material to be cut. This chapter analyzes interactions between food products and ultrasonic cutting tools and relates these interactions with physical and chemical product properties as well as with processing parameters like cutting velocity, ultrasonic amplitude and frequency, and tool design.

  10. Cloning and expression of full-length cDNA encoding human vitamin D receptor

    Energy Technology Data Exchange (ETDEWEB)

    Baker, A.R.; McDonnell, D.P.; Hughes, M.; Crisp, T.M.; Mangelsdorf, D.J.; Haussler, M.R.; Pike, J.W.; Shine, J.; O' Malley, B.W. (California Biotechnology Inc., Mountain View (USA))

    1988-05-01

    Complementary DNA clones encoding the human vitamin D receptor have been isolated from human intestine and T47D cell cDNA libraries. The nucleotide sequence of the 4605-base pair (bp) cDNA includes a noncoding leader sequence of 115 bp, a 1281-bp open reading frame, and 3209 bp of 3{prime} noncoding sequence. Two polyadenylylation signals, AATAAA, are present 25 and 70 bp upstream of the poly(A) tail, respectively. RNA blot hybridization indicates a single mRNA species of {approx} 4600 bp. Transfection of the cloned sequences into COS-1 cells results in the production of a single receptor species indistinguishable from the native receptor. Sequence comparisons demonstrate that the vitamin D receptor belongs to the steroid-receptor gene family and is closest in size and sequence to another member of this family, the thyroid hormone receptor.

  11. Saving Seal Cutting

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    On April 20, the graduation ceremony of China’s seal-cutting art postgraduates and visiting experts from the Institute of Seal Cutting Art under the China Art Academy was held in Beijing. On the same day, the exhibition of the works of the teachers and graduates of the institute was also held.

  12. Cutting Class Harms Grades

    Science.gov (United States)

    Taylor, Lewis A., III

    2012-01-01

    An accessible business school population of undergraduate students was investigated in three independent, but related studies to determine effects on grades due to cutting class and failing to take advantage of optional reviews and study quizzes. It was hypothesized that cutting classes harms exam scores, attending preexam reviews helps exam…

  13. Four Families Characte rized by Culture

    Institute of Scientific and Technical Information of China (English)

    1994-01-01

    in recent years, such characteristic families as the family of Beijing opera, the family of calligraphy and the family of paper-cutting have emerged in the Beixinqiao Neighborhood of Dongcheng District, in Beijing.

  14. Fundamentals of cutting.

    Science.gov (United States)

    Williams, J G; Patel, Y

    2016-06-06

    The process of cutting is analysed in fracture mechanics terms with a view to quantifying the various parameters involved. The model used is that of orthogonal cutting with a wedge removing a layer of material or chip. The behaviour of the chip is governed by its thickness and for large radii of curvature the chip is elastic and smooth cutting occurs. For smaller thicknesses, there is a transition, first to plastic bending and then to plastic shear for small thicknesses and smooth chips are formed. The governing parameters are tool geometry, which is principally the wedge angle, and the material properties of elastic modulus, yield stress and fracture toughness. Friction can also be important. It is demonstrated that the cutting process may be quantified via these parameters, which could be useful in the study of cutting in biology.

  15. Construction of cDNA Library from Populus euphratica

    Institute of Scientific and Technical Information of China (English)

    Yu Guangjun; Wang Yiqin; Shen Xin

    2003-01-01

    In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on the beads. The double strand cDNA were released from magnetic beads by digestion with NotI, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA were directionally cloned intoλExcell vector. The results show that the primary titer of the cDNA library is 7.46×106 pfu per mL and the packaging efficiency reaches 1.47×107 recombinants per μg DNA. λDNA extracted from two clones of plaque were digested by EcoR I and NotI, both of the clones contained inserts larger than 900 bp. These results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed.

  16. Online Cake Cutting

    CERN Document Server

    Walsh, Toby

    2010-01-01

    We propose an online form of the cake cutting problem. This models situations where players arrive and depart during the process of dividing a resource. We show that well known fair division procedures like cut-and-choose and the Dubins-Spanier moving knife procedure can be adapted to apply to such online problems. We propose some desirable properties that online cake cutting procedures might possess like online forms of proportionality and envy-freeness, and identify which properties are in fact possessed by the different online cake procedures.

  17. Experimental testing of exchangeable cutting inserts cutting ability

    OpenAIRE

    Čep, Robert; Janásek, Adam; Čepová Lenka; Petrů, Jana; Ivo HLAVATÝ; Car, Zlatan; Hatala, Michal

    2013-01-01

    The article deals with experimental testing of the cutting ability of exchangeable cutting inserts. Eleven types of exchangeable cutting inserts from five different manufacturers were tested. The tested cutting inserts were of the same shape and were different especially in material and coating types. The main aim was both to select a suitable test for determination of the cutting ability of exchangeable cutting inserts and to design such testing procedure that could make it possible...

  18. Testing Of Choiced Ceramics Cutting Tools At Irregular Interrupted Cut

    Science.gov (United States)

    Kyncl, Ladislav; Malotová, Šárka; Nováček, Pavel; Nicielnik, Henryk; Šoková, Dagmar; Hemžský, Pavel; Pitela, David; Holubjak, Jozef

    2015-12-01

    This article discusses the test of removable ceramic cutting inserts during machining irregular interrupted cut. Tests were performed on a lathe, with the preparation which simulated us the interrupted cut. By changing the number of plates mounted in a preparation it simulate us a regular or irregular interrupted cut. When with four plates it was regular interrupted cut, the remaining three variants were already irregular cut. It was examined whether it will have the irregular interrupted cutting effect on the insert and possibly how it will change life of inserts during irregular interrupted cut (variable delay between shocks).

  19. Giardia canis: ultrastructural analysis of G. canis trophozoites transfected with full length G. canis virus cDNA transcripts

    Science.gov (United States)

    Giardia canis virus (GCV) is a double-stranded RNA (dsRNA) virus of the family Totiviridae. In this study, the full-length cDNA of the G. canis virus was constructed in pPoly2/sfinot vector and RNA was transcribed in vitro. Virus-free G. canis trophozoites were transfected with in vitro transcribed ...

  20. Determination of cDNA and genomic DNA sequences of hevamine, a chitinase from the rubber tree Hevea brasiliensis

    NARCIS (Netherlands)

    Bokma, E; Spiering, M; Chow, KS; Mulder, PPMFA; Subroto, T; Beintema, JJ

    2001-01-01

    Hevamine is a chitinase from the rubber tree Hevea brasiliensis and belongs to the family 18 glycosyl hydrolases. This paper describes the cloning of hevamine DNA and cDNA sequences. Hevamine contains a signal peptide at the N-terminus and a putative vacuolar targeting sequence at the C-terminus whi

  1. Short-cut math

    CERN Document Server

    Kelly, Gerard W

    1984-01-01

    Clear, concise compendium of about 150 time-saving math short-cuts features faster, easier ways to add, subtract, multiply, and divide. Each problem includes an explanation of the method. No special math ability needed.

  2. Cutting Cakes Correctly

    CERN Document Server

    Hill, Theodore P

    2008-01-01

    Without additional hypotheses, Proposition 7.1 in Brams and Taylor's book "Fair Division" (Cambridge University Press, 1996) is false, as are several related Pareto-optimality theorems of Brams, Jones and Klamler in their 2006 cake-cutting paper.

  3. Laser cutting system

    Science.gov (United States)

    Dougherty, Thomas J

    2015-03-03

    A workpiece cutting apparatus includes a laser source, a first suction system, and a first finger configured to guide a workpiece as it moves past the laser source. The first finger includes a first end provided adjacent a point where a laser from the laser source cuts the workpiece, and the first end of the first finger includes an aperture in fluid communication with the first suction system.

  4. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mark Pickell; Len Volk; Mike Volk; Evren Ozbayoglu; Lei Zhou

    2002-07-30

    This is the fourth quarterly progress report for Year-3 of the ACTS Project. It includes a review of progress made in: (1) Flow Loop construction and development and (2) research tasks during the period of time between April 1, 2002 and June 30, 2002. This report presents a review of progress on the following specific tasks: (a) Design and development of an Advanced Cuttings Transport Facility (Task 3: Addition of a Cuttings Injection/Separation System), (b) Research project (Task 6): ''Study of Cuttings Transport with Foam Under LPAT Conditions (Joint Project with TUDRP)''; (c) Research project (Task 9b): ''Study of Foam Flow Behavior Under EPET Conditions''; (d) Research project (Task 10): ''Study of Cuttings Transport with Aerated Mud Under Elevated Pressure and Temperature Conditions''; (e) Research on three instrumentation tasks to measure: Cuttings concentration and distribution in a flowing slurry (Task 11), Foam texture while transporting cuttings. (Task 12), and Viscosity of Foam under EPET (Task 9b); (f) Development of a Safety program for the ACTS Flow Loop. Progress on a comprehensive safety review of all flow-loop components and operational procedures. (Task 1S); (g) Activities towards technology transfer and developing contacts with Petroleum and service company members, and increasing the number of JIP members.

  5. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mark Pickell; Len Volk; Mike Volk; Lei Zhou; Zhu Chen; Crystal Redden; Aimee Washington

    2003-01-30

    This is the second quarterly progress report for Year-4 of the ACTS Project. It includes a review of progress made in: (1) Flow Loop construction and development and (2) research tasks during the period of time between October 1, 2002 and December 30, 2002. This report presents a review of progress on the following specific tasks. (a) Design and development of an Advanced Cuttings Transport Facility Task 3: Addition of a Cuttings Injection/Separation System, Task 4: Addition of a Pipe Rotation System. (b) New research project (Task 9b): ''Development of a Foam Generator/Viscometer for Elevated Pressure and Elevated Temperature (EPET) Conditions''. (d) Research project (Task 10): ''Study of Cuttings Transport with Aerated Mud Under Elevated Pressure and Temperature Conditions''. (e) Research on three instrumentation tasks to measure: Cuttings concentration and distribution in a flowing slurry (Task 11), Foam texture while transporting cuttings. (Task 12), and Viscosity of Foam under EPET (Task 9b). (f) New Research project (Task 13): ''Study of Cuttings Transport with Foam under Elevated Pressure and Temperature Conditions''. (g) Development of a Safety program for the ACTS Flow Loop. Progress on a comprehensive safety review of all flow-loop components and operational procedures. (Task 1S). (h) Activities towards technology transfer and developing contacts with Petroleum and service company members, and increasing the number of JIP members.

  6. Cloning of a cDNA for steroid sulfatase: frequent occurrence of gene deletions in patients with recessive X chromosome-linked ichthyosis

    Energy Technology Data Exchange (ETDEWEB)

    Bonifas, J.M.; Morley, B.J.; Oakey, R.E.; Kan, Y.W.; Epstein, E.J. Jr.

    1987-12-01

    A human steroid sulfatase cDNA 2.4 kilobases long was isolated from a human placental lambda gt11 cDNA expression library. The library was screened with monospecific rabbit antibodies elicited by injection of steroid sulfatase protein purified from human placentas. Hybridization of the cDNA with EcoRI-digested genomic DNA indicated that patients from 14 of 15 apparently unrelated families have gross deletions of the gene for steroid sulfatase. One patient had genomic DNA fragments that were identical to those from normal individuals, indicating the absence of any major deletions as the cause of his lack of steroid sulfatase enzyme activity.

  7. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mengjiao Yu; Ramadan Ahmed; Mark Pickell; Len Volk; Lei Zhou; Zhu Chen; Aimee Washington; Crystal Redden

    2003-09-30

    The Quarter began with installing the new drill pipe, hooking up the new hydraulic power unit, completing the pipe rotation system (Task 4 has been completed), and making the SWACO choke operational. Detailed design and procurement work is proceeding on a system to elevate the drill-string section. The prototype Foam Generator Cell has been completed by Temco and delivered. Work is currently underway to calibrate the system. Literature review and preliminary model development for cuttings transportation with polymer foam under EPET conditions are in progress. Preparations for preliminary cuttings transport experiments with polymer foam have been completed. Two nuclear densitometers were re-calibrated. Drill pipe rotation system was tested up to 250 RPM. Water flow tests were conducted while rotating the drill pipe up to 100 RPM. The accuracy of weight measurements for cuttings in the annulus was evaluated. Additional modifications of the cuttings collection system are being considered in order to obtain the desired accurate measurement of cuttings weight in the annular test section. Cutting transport experiments with aerated fluids are being conducted at EPET, and analyses of the collected data are in progress. The printed circuit board is functioning with acceptable noise level to measure cuttings concentration at static condition using ultrasonic method. We were able to conduct several tests using a standard low pass filter to eliminate high frequency noise. We tested to verify that we can distinguish between different depths of sand in a static bed of sand. We tested with water, air and a mix of the two mediums. Major modifications to the DTF have almost been completed. A stop-flow cell is being designed for the DTF, the ACTF and Foam Generator/Viscometer which will allow us to capture bubble images without the need for ultra fast shutter speeds or microsecond flash system.

  8. Improved cutting performance in high power laser cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    2003-01-01

    Recent results in high power laser cutting especially with focus on cutting of mild grade steel types for shipbuilding are described.......Recent results in high power laser cutting especially with focus on cutting of mild grade steel types for shipbuilding are described....

  9. Improved cutting performance in high power laser cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    2003-01-01

    Recent results in high power laser cutting especially with focus on cutting of mild grade steel types for shipbuilding are described.......Recent results in high power laser cutting especially with focus on cutting of mild grade steel types for shipbuilding are described....

  10. Cloning of human tumor necrosis factor (TNF) receptor cDNA and expression of recombinant soluble TNF-binding protein.

    OpenAIRE

    Gray, P W; Barrett, K; Chantry, D; Turner, M.; Feldmann, M

    1990-01-01

    The cDNA for one of the receptors for human tumor necrosis factor (TNF) has been isolated. This cDNA encodes a protein of 455 amino acids that is divided into an extracellular domain of 171 residues and a cytoplasmic domain of 221 residues. The extracellular domain has been engineered for expression in mammalian cells, and this recombinant derivative binds TNF alpha with high affinity and inhibits its cytotoxic activity in vitro. The TNF receptor exhibits similarity with a family of cell surf...

  11. Determination of cut front position in laser cutting

    Science.gov (United States)

    Pereira, M.; Thombansen, U.

    2016-07-01

    Laser cutting has a huge importance to manufacturing industry. Laser cutting machines operate with fixed technological parameters and this does not guarantee the best productivity. The adjustment of the cutting parameters during operation can improve the machine performance. Based on a coaxial measuring device it is possible to identify the cut front position during the cutting process. This paper describes the data analysis approach used to determine the cut front position for different feed rates. The cut front position was determined with good resolution, but improvements are needed to make the whole process more stable.

  12. Characterization of the porcine carboxypeptidase E cDNA

    DEFF Research Database (Denmark)

    Hreidarsdôttir, G.E.; Cirera, Susanna; Fredholm, Merete

    2007-01-01

    the sequence of the cDNA for the porcine CPE gene including all the coding region and the 3'-UTR region was generated. Comparisons with bovine, human, mouse, and rat CPE cDNA sequences showed that the coding regions of the gene are highly conserved both at the nucleotide and at the amino acid level. A very low...

  13. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mark Pickell; Len Volk, Mike Volk; Lei Zhou; Zhu Chen; Crystal Redden; Aimee Washington

    2002-10-30

    This is the first quarterly progress report for Year-4 of the ACTS Project. It includes a review of progress made in: (1) Flow Loop construction and development and (2) research tasks during the period of time between July 1, 2002 and Sept. 30, 2002. This report presents a review of progress on the following specific tasks: (a) Design and development of an Advanced Cuttings Transport Facility Task 3: Addition of a Cuttings Injection/Separation System, Task 4: Addition of a Pipe Rotation System, (b) New Research project (Task 9b): ''Development of a Foam Generator/Viscometer for Elevated Pressure and Elevated Temperature (EPET) Conditions'', (d) Research project (Task 10): ''Study of Cuttings Transport with Aerated Mud Under Elevated Pressure and Temperature Conditions'', (e) Research on three instrumentation tasks to measure: Cuttings concentration and distribution in a flowing slurry (Task 11), Foam texture while transporting cuttings (Task 12), Viscosity of Foam under EPET (Task 9b). (f) Development of a Safety program for the ACTS Flow Loop. Progress on a comprehensive safety review of all flow-loop components and operational procedures. (Task 1S). (g) Activities towards technology transfer and developing contacts with Petroleum and service company members, and increasing the number of JIP members.

  14. PvuII RFLP detected by a human. beta. ADH cDNA probe

    Energy Technology Data Exchange (ETDEWEB)

    Parsian, A.; Burgess, A.K.; Khan, M.A.; Devor, E.J. (Washington Univ. School of Medicine, St. Louis, MO (USA))

    1989-12-11

    A 0.97 kb cDNA (ADH12) fragment encoding human exons 7, 8, 9 of the ADH{sub 2} gene was isolated from an adult human liver cDNA library. The insert can be excised by Pst I digestion. Pvu II identifies a two-allele polymorphism with bands at 4.4 kb (A{sub 1}) and 3.0 kb (A{sub 2}) and invariant bands at 5.1, 4.0, 2.8, and 2.3 kb. It was localized on Chromosome 4q21-q25 by in situ hybridization. Co-dominant segregation was observed in 18 informative families.

  15. Dealing with Cuts (For Parents)

    Science.gov (United States)

    ... For Kids For Parents MORE ON THIS TOPIC Cellulitis First Aid: Cuts Staph Infections Bites and Scratches First Aid: Falls First Aid: ... Out Cuts, Scratches, and Abrasions What's a Scab? Cellulitis Cuts, Scratches, and Scrapes Staph Infections Dealing With Cuts and Wounds Contact Us Print ...

  16. Theoretical Models for Orthogonal Cutting

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo

    This review of simple models for orthogonal cutting was extracted from: “L. De Chiffre: Metal Cutting Mechanics and Applications, D.Sc. Thesis, Technical University of Denmark, 1990.”......This review of simple models for orthogonal cutting was extracted from: “L. De Chiffre: Metal Cutting Mechanics and Applications, D.Sc. Thesis, Technical University of Denmark, 1990.”...

  17. Theoretical Models for Orthogonal Cutting

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo

    This review of simple models for orthogonal cutting was extracted from: “L. De Chiffre: Metal Cutting Mechanics and Applications, D.Sc. Thesis, Technical University of Denmark, 1990.”......This review of simple models for orthogonal cutting was extracted from: “L. De Chiffre: Metal Cutting Mechanics and Applications, D.Sc. Thesis, Technical University of Denmark, 1990.”...

  18. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Gerald Kane; Mark Pickell; Len Volk; Mike Volk; Barkim Demirdal; Affonso Lourenco; Evren Ozbayoglu; Paco Vieira; Lei Zhou

    2000-01-30

    This is the second quarterly progress report for Year 2 of the ACTS project. It includes a review of progress made in Flow Loop development and research during the period of time between Oct 1, 2000 and December 31, 2000. This report presents a review of progress on the following specific tasks: (a) Design and development of an Advanced Cuttings Transport Facility (Task 2: Addition of a foam generation and breaker system), (b) Research project (Task 6): ''Study of Cuttings Transport with Foam Under LPAT Conditions (Joint Project with TUDRP)'', (c) Research project (Task 7): ''Study of Cuttings Transport with Aerated Muds Under LPAT Conditions (Joint Project with TUDRP)'', (d) Research project (Task 8): ''Study of Flow of Synthetic Drilling Fluids Under Elevated Pressure and Temperature Conditions'', (e) Research project (Task 9): ''Study of Foam Flow Behavior Under EPET Conditions'', (f) Research project (Task 10): ''Study of Cuttings Transport with Aerated Mud Under Elevated Pressure and Temperature Conditions'', (g) Research on instrumentation tasks to measure: Cuttings concentration and distribution in a flowing slurry (Task 11), and Foam properties while transporting cuttings. (Task 12), (h) Development of a Safety program for the ACTS Flow Loop. Progress on a comprehensive safety review of all flow-loop components and operational procedures. (Task 1S). (i) Activities towards technology transfer and developing contacts with Petroleum and service company members, and increasing the number of JIP members. The tasks Completed During This Quarter are Task 7 and Task 8.

  19. The perfect cut

    DEFF Research Database (Denmark)

    Scozzafava, G.; Mueller Loose, Simone; Corsi, A.

    (organic, standard, GMO free). The cross-price elasticity provides insights to which degree different cuts compete against each other from a consumer perspective and how price premiums can be achieved by producers and marketers with certification and labeling strategies. The paper will also provide...... other from the consumer perspective dependent on price, intrinsic and extrinsic product characteristics as well as intended usage. So far, there is limited knowledge about optimal marketing and pricing of meat cuts simultaneously offered at the retail shelf. Results from an online choice experiment...

  20. Molecular cloning and sequence analysis of growth hormone cDNA of Neotropical freshwater fish Pacu (Piaractus mesopotamicus

    Directory of Open Access Journals (Sweden)

    Janeth Silva Pinheiro

    2008-01-01

    Full Text Available RT-PCR was used for amplifying Piaractus mesopotamicus growth hormone (GH cDNA obtained from mRNA extracted from pituitary cells. The amplified fragment was cloned and the complete cDNA sequence was determined. The cloned cDNA encompassed a sequence of 543 nucleotides that encoded a polypeptide of 178 amino acids corresponding to mature P. mesopotamicus GH. Comparison with other GH sequences showed a gap of 10 amino acids localized in the N terminus of the putative polypeptide of P. mesopotamicus. This same gap was also observed in other members of the family. Neighbor-joining tree analysis with GH sequences from fishes belonging to different taxonomic groups placed the P. mesopotamicus GH within the Otophysi group. To our knowledge, this is the first GH sequence of a Neotropical characiform fish deposited in GenBank.

  1. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Stefan Miska; Troy Reed; Ergun Kuru

    2004-09-30

    The Advanced Cuttings Transport Study (ACTS) was a 5-year JIP project undertaken at the University of Tulsa (TU). The project was sponsored by the U.S. Department of Energy (DOE) and JIP member companies. The objectives of the project were: (1) to develop and construct a new research facility that would allow three-phase (gas, liquid and cuttings) flow experiments under ambient and EPET (elevated pressure and temperature) conditions, and at different angle of inclinations and drill pipe rotation speeds; (2) to conduct experiments and develop a data base for the industry and academia; and (3) to develop mechanistic models for optimization of drilling hydraulics and cuttings transport. This project consisted of research studies, flow loop construction and instrumentation development. Following a one-year period for basic flow loop construction, a proposal was submitted by TU to the DOE for a five-year project that was organized in such a manner as to provide a logical progression of research experiments as well as additions to the basic flow loop. The flow loop additions and improvements included: (1) elevated temperature capability; (2) two-phase (gas and liquid, foam etc.) capability; (3) cuttings injection and removal system; (4) drill pipe rotation system; and (5) drilling section elevation system. In parallel with the flow loop construction, hydraulics and cuttings transport studies were preformed using drilling foams and aerated muds. In addition, hydraulics and rheology of synthetic drilling fluids were investigated. The studies were performed under ambient and EPET conditions. The effects of temperature and pressure on the hydraulics and cuttings transport were investigated. Mechanistic models were developed to predict frictional pressure loss and cuttings transport in horizontal and near-horizontal configurations. Model predictions were compared with the measured data. Predominantly, model predictions show satisfactory agreements with the measured data. As a

  2. "Kid Cuts" by Broderbund.

    Science.gov (United States)

    Martin, Ron

    1994-01-01

    Describes "Kid Cuts," an arts and crafts computer software program for students in prekindergarten through sixth grade that provides 22 activities in 6 curriculum areas. An example is given of an activity for kindergarten and first graders related to counting that includes library media skills objectives and mathematics objectives. (LRW)

  3. Cutting Cakes Carefully

    Science.gov (United States)

    Hill, Theodore P.; Morrison, Kent E.

    2010-01-01

    This paper surveys the fascinating mathematics of fair division, and provides a suite of examples using basic ideas from algebra, calculus, and probability which can be used to examine and test new and sometimes complex mathematical theories and claims involving fair division. Conversely, the classical cut-and-choose and moving-knife algorithms…

  4. Simultaneous Cake Cutting

    DEFF Research Database (Denmark)

    Balkanski, Eric; Branzei, Simina; Kurokawa, David;

    2014-01-01

    We introduce the simultaneous model for cake cutting (the fair allocation of a divisible good), in which agents simultaneously send messages containing a sketch of their preferences over the cake. We show that this model enables the computation of divisions that satisfy proportionality — a popular...

  5. Cutting Cakes Carefully

    Science.gov (United States)

    Hill, Theodore P.; Morrison, Kent E.

    2010-01-01

    This paper surveys the fascinating mathematics of fair division, and provides a suite of examples using basic ideas from algebra, calculus, and probability which can be used to examine and test new and sometimes complex mathematical theories and claims involving fair division. Conversely, the classical cut-and-choose and moving-knife algorithms…

  6. Kids Who Cut.

    Science.gov (United States)

    Coy, Doris Rhea; Simpson, Chris

    2002-01-01

    Regardless of whether it is cutting, burning or some other form of self-harm, self-injury is a serious problem requiring serious solutions. This article reviews the various types of self-harm, descriptions of self-mutilators, common myths about self-mutilation, and effective treatment methods. (GCP)

  7. cDNA cloning and sequencing of ostrich Growth hormone

    Directory of Open Access Journals (Sweden)

    Doosti Abbas

    2012-01-01

    Full Text Available In recent years, industrial breeding of ostrich (Struthio camelus has been widely developed in Iran. Growth hormone (GH is a peptide hormone that stimulates growth and cell reproduction in different animals. The aim of this study was to clone and sequence the ostrich growth hormone gene in E. coli, done for the first time in Iran. The cDNA that encodes ostrich growth hormone was isolated from total mRNA of the pituitary gland and amplified by RT-PCR using GH specific PCR primers. Then GH cDNA was cloned by T/A cloning technique and the construct was transformed into E. coli. Finally, GH cDNA sequence was submitted to the GenBank (Accession number: JN559394. The results of present study showed that GH cDNA was successfully cloned in E. coli. Sequencing confirmed that GH cDNA was cloned and that the length of ostrich GH cDNA was 672 bp; BLAST search showed that the sequence of growth hormone cDNA of the ostrich from Iran has 100% homology with other records existing in GenBank.

  8. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Gerald Kane; Mark Pickell; Len Volk; Mike Volk; Barkim Demirdal; Affonso Lourenco; Evren Ozbayoglu; Paco Vieira

    2000-10-30

    This is the first quarterly progress report for Year 2 of the ACTS project. It includes a review of progress made in Flow Loop development and research during the period of time between July 14, 2000 and September 30, 2000. This report presents information on the following specific tasks: (a) Progress in Advanced Cuttings Transport Facility design and development (Task 2), (b) Progress on research project (Task 8): ''Study of Flow of Synthetic Drilling Fluids Under Elevated Pressure and Temperature Conditions'', (c) Progress on research project (Task 6): ''Study of Cuttings Transport with Foam Under LPAT Conditions (Joint Project with TUDRP)'', (d) Progress on research project (Task 7): ''Study of Cuttings Transport with Aerated Muds Under LPAT Conditions (Joint Project with TUDRP)'', (e) Progress on research project (Task 9): ''Study of Foam Flow Behavior Under EPET Conditions'', (f) Initiate research on project (Task 10): ''Study of Cuttings Transport with Aerated Mud Under Elevated Pressure and Temperature Conditions'', (g) Progress on instrumentation tasks to measure: Cuttings concentration and distribution (Tasks 11), and Foam properties (Task 12), (h) Initiate a comprehensive safety review of all flow-loop components and operational procedures. Since the previous Task 1 has been completed, we will now designate this new task as: (Task 1S). (i) Activities towards technology transfer and developing contacts with Petroleum and service company members, and increasing the number of JIP members.

  9. Molecular cloning of lupin leghemoglobin cDNA

    DEFF Research Database (Denmark)

    Konieczny, A; Jensen, E O; Marcker, K A

    1987-01-01

    Poly(A)+ RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences...... its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+ RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules. Udgivelsesdato: 1987-null...

  10. Molecular cloning of lupin leghemoglobin cDNA

    DEFF Research Database (Denmark)

    Konieczny, A; Jensen, E O; Marcker, K A

    1987-01-01

    Poly(A)+ RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences...... its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+ RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules. Udgivelsesdato: 1987-null...

  11. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mark Pickell; Len Volk; Mike Volk; Lei Zhou; Zhu Chen; Crystal Redden; Aimee Washington

    2003-04-30

    Experiments on the flow loop are continuing. Improvements to the software for data acquisition are being made as additional experience with three-phase flow is gained. Modifications are being made to the Cuttings Injection System in order to improve control and the precision of cuttings injection. The design details for a drill-pipe Rotation System have been completed. A US Patent was filed on October 28, 2002 for a new design for an instrument that can generate a variety of foams under elevated pressures and temperatures and then transfer the test foam to a viscometer for measurements of viscosity. Theoretical analyses of cuttings transport phenomena based on a layered model is under development. Calibrations of two nuclear densitometers have been completed. Baseline tests have been run to determine wall roughness in the 4 different tests sections (i.e. 2-in, 3-in, 4-in pipes and 5.76-in by 3.5-in annulus) of the flow loop. Tests have also been conducted with aerated fluids at EPET conditions. Preliminary experiments on the two candidate aqueous foam formulations were conducted which included rheological tests of the base fluid and foam stability reports. These were conducted after acceptance of the proposal on the Study of Cuttings Transport with Foam Under Elevated Pressure and Elevated Temperature Conditions. Preparation of a test matrix for cuttings-transport experiments with foam in the ACTF is also under way. A controller for instrumentation to measure cuttings concentration and distribution has been designed that can control four transceivers at a time. A prototype of the control circuit board was built and tested. Tests showed that there was a problem with radiated noise. AN improved circuit board was designed and sent to an external expert to verify the new design. The new board is being fabricated and will first be tested with static water and gravel in an annulus at elevated temperatures. A series of viscometer tests to measure foam properties have

  12. cDNA cloning and characterization of a mannose-binding lectin from Zingiber officinale Roscoe (ginger) rhizomes

    Indian Academy of Sciences (India)

    Zhonghai Chen; Guoyin Kai; Xiaojun Liu; Juan Lin; Xiaofen Sun; Kexuan Tang

    2005-03-01

    Using RNA extracted from Zingiber officinale rhizomes and primers designed according to the conservative regions of monocot mannose-binding lectins, the full-length cDNA of Z. officinale agglutinin (ZOA) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of zoa was 746 bp and contained a 510 bp open reading frame (ORF) encoding a lectin precursor of 169 amino acids with a signal peptide. ZOA was a mannose-binding lectin with three typical mannose-binding sites (QDNY). Semi-quantitative RT-PCR analysis revealed that zoa expressed in all the tested tissues of Z. officinale including leaf, root and rhizome, suggesting it to be a constitutively expressing form. ZOA protein was successfully expressed in Escherichia coli with the molecular weight expected. To our knowledge, this is the first mannose-binding lectin cDNA cloned from the family Zingiberaceae. Our results demonstrate that monocot mannose-binding lectins also occur within the family Zingiberaceae.

  13. Method for construction of normalized cDNA libraries

    Science.gov (United States)

    Soares, Marcelo B.; Efstratiadis, Argiris

    1998-01-01

    This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to appropriate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library. This invention also provides normalized cDNA libraries generated by the above-described method and uses of the generated libraries.

  14. Cutting forces during turning with variable depth of cut

    Directory of Open Access Journals (Sweden)

    M. Sadílek

    2016-03-01

    The proposed research for the paper is an experimental work – measuring cutting forces and monitoring of the tool wear on the cutting edge. It compares the turning where standard roughing cycle is used and the turning where the proposed roughing cycle with variable depth of cut is applied.

  15. Application of restriction display PCR technique in the preparation of cDNA microarray probes

    Institute of Scientific and Technical Information of China (English)

    Zhao-Hui Sun; Wen-Li Ma; Bao Zhang; Yi-Fei Peng; Wen-Ling Zheng

    2005-01-01

    AIM: To develop a simplified and efficient method for the preparation of hepatitis C virus (HCV) cDNA microarray probes.METHODS: With the technique of restriction display PCR (RD-PCR), restriction enzyme Sau3A I was chosen to digest the full-length HCV cDNAs. The products were classified and re-amplified by RD-PCR. We separated the differential genes by polyacrylamide gel electrophoresis and silver staining. Single bands cut out from the polyacrylamide gel were isolated. The third-round PCR was performed using the single bands as PCR template.The RD-PCR fragments were purified and cloned into the pMD18-T vector. The recombinant plasmids were extracted from positive clones, and the target gene fragments were sequenced. The cDNA microarray was prepared by spotting RD-PCR products to the surface of amino-modified glass slides using a robot. We validated the detection of microarray by hybridization and sequence analysis.RESULTS: A total of 24 different cDNA fragments ranging from 200 to 800 bp were isolated and sequenced,which were the specific gene fragments of HCV. These fragments could be further used as probes in microarray preparation. The diagnostic capability of the microarray was evaluated after the washing and scanning steps. The results of hybridization and sequence analysis showed that the specificity, sensitivity, accuracy, reproducibility,and linearity in detecting HCV RNA were satisfactory.CONCLUSION: The RD-PCR technique is of great value in obtaining a large number of size-comparable gene probes, which provides a speedy protocol in generating probes for the preparation of microarrays. Microarray prepared as such could be further optimized and applied in the clinical diagnosis of HCV.

  16. Cutting Out Continuations

    DEFF Research Database (Denmark)

    Bahr, Patrick; Hutton, Graham

    2016-01-01

    In the field of program transformation, one often transforms programs into continuation-passing style to make their flow of control explicit, and then immediately removes the resulting continuations using defunctionalisation to make the programs first-order. In this article, we show how these two...... transformations can be fused together into a single transformation step that cuts out the need to first introduce and then eliminate continuations. Our approach is calculational, uses standard equational reasoning techniques, and is widely applicable....

  17. Manual bamboo cutting tool.

    Science.gov (United States)

    Bezerra, Mariana Pereira; Correia, Walter Franklin Marques; da Costa Campos, Fabio Ferreira

    2012-01-01

    The paper presents the development of a cutting tool guide, specifically for the harvest of bamboo. The development was made based on precepts of eco-design and ergonomics, for prioritizing the physical health of the operator and the maintenance of the environment, as well as meet specific requirements of bamboo. The main goal is to spread the use of bamboo as construction material, handicrafts, among others, from a handy, easy assembly and material available tool.

  18. Making the cut

    OpenAIRE

    Millard, Chris

    2013-01-01

    ‘Deliberate self-harm’, ‘self-mutilation’ and ‘self-injury’ are just some of the terms used to describe one of the most prominent issues in British mental health policy in recent years. This article demonstrates that contemporary literature on ‘self-harm’ produces this phenomenon (to varying extents) around two key characteristics. First, this behaviour is predominantly performed by those identified as female. Second, this behaviour primarily involves cutting the skin. These constitutive char...

  19. Soluble oil cutting fluid

    Energy Technology Data Exchange (ETDEWEB)

    Rawlinson, A.P.; White, J.

    1987-06-23

    A soluble oil, suitable when diluted with water, for use as a cutting fluid comprises an alkali or alkaline-earth metal alkyl benzene sulphonate, a fatty acid diethanolamide, a mixed alkanolamine borate, a polyisobutenesuccinimide and a major proportion of mineral oil. The soluble oil is relatively stable without the need for a conventional coupling agent and some soluble oil emulsions are bio-static even though conventional biocides are not included.

  20. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Ergun Kuru; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Gerald Kane; Mark Pickell; Len Volk; Mike Volk; Barkim Demirdal; Affonso Lourenco; Evren Ozbayoglu; Paco Vieira; Neelima Godugu

    2000-07-30

    ACTS flow loop is now operational under elevated pressure and temperature. Currently, experiments with synthetic based drilling fluids under pressure and temperature are being conducted. Based on the analysis of Fann 70 data, empirical correlations defining the shear stress as a function of temperature, pressure and the shear rate have been developed for Petrobras synthetic drilling fluids. PVT equipment has been modified for testing Synthetic oil base drilling fluids. PVT tests with Petrobras Synthetic base mud have been conducted and results are being analyzed Foam flow experiments have been conducted and the analysis of the data has been carried out to characterize the rheology of the foam. Comparison of pressure loss prediction from the available foam hydraulic models and the test results has been made. Cuttings transport experiments in horizontal annulus section have been conducted using air, water and cuttings. Currently, cuttings transport tests in inclined test section are being conducted. Foam PVT analysis tests have been conducted. Foam stability experiments have also been conducted. Effects of salt and oil concentration on the foam stability have been investigated. Design of ACTS flow loop modification for foam and aerated mud flow has been completed. A flow loop operation procedure for conducting foam flow experiments under EPET conditions has been prepared Design of the lab-scale flow loop for dynamic foam characterization and cuttings monitoring instrumentation tests has been completed. The construction of the test loop is underway. As part of the technology transport efforts, Advisory Board Meeting with ACTS-JIP industry members has been organized on May 13, 2000.

  1. Lectin cDNA and transgenic plants derived therefrom

    Science.gov (United States)

    Raikhel, Natasha V.

    1994-01-04

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties. GOVERNMENT RIGHTS This application was funded under Department of Energy Contract DE-AC02-76ER01338. The U.S. Government has certain rights under this application and any patent issuing thereon.

  2. Differential cDNA cloning by enzymatic degrading subtraction (EDS).

    OpenAIRE

    1994-01-01

    We describe a new method, called enzymatic degrading subtraction (EDS), for the construction of subtractive libraries from PCR amplified cDNA. The novel features of this method are that i) the tester DNA is blocked by thionucleotide incorporation; ii) the rate of hybridization is accelerated by phenol-emulsion reassociation; and iii) the driver cDNA and hybrid molecules are enzymatically removed by digestion with exonucleases III and VII rather than by physical partitioning. We demonstrate th...

  3. Constructing and detecting a cDNA library for mites.

    Science.gov (United States)

    Hu, Li; Zhao, YaE; Cheng, Juan; Yang, YuanJun; Li, Chen; Lu, ZhaoHui

    2015-10-01

    RNA extraction and construction of complementary DNA (cDNA) library for mites have been quite challenging due to difficulties in acquiring tiny living mites and breaking their hard chitin. The present study is to explore a better method to construct cDNA library for mites that will lay the foundation on transcriptome and molecular pathogenesis research. We selected Psoroptes cuniculi as an experimental subject and took the following steps to construct and verify cDNA library. First, we combined liquid nitrogen grinding with TRIzol for total RNA extraction. Then, switching mechanism at 5' end of the RNA transcript (SMART) technique was used to construct full-length cDNA library. To evaluate the quality of cDNA library, the library titer and recombination rate were calculated. The reliability of cDNA library was detected by sequencing and analyzing positive clones and genes amplified by specific primers. The results showed that the RNA concentration was 836 ng/μl and the absorbance ratio at 260/280 nm was 1.82. The library titer was 5.31 × 10(5) plaque-forming unit (PFU)/ml and the recombination rate was 98.21%, indicating that the library was of good quality. In the 33 expressed sequence tags (ESTs) of P. cuniculi, two clones of 1656 and 1658 bp were almost identical with only three variable sites detected, which had an identity of 99.63% with that of Psoroptes ovis, indicating that the cDNA library was reliable. Further detection by specific primers demonstrated that the 553-bp Pso c II gene sequences of P. cuniculi had an identity of 98.56% with those of P. ovis, confirming that the cDNA library was not only reliable but also feasible.

  4. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Ergun Kuru; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Gerald Kane; Len Volk; Mark Pickell; Evren Ozbayoglu; Barkim Demirdal; Paco Vieira; Affonso Lourenco

    1999-10-15

    This report includes a review of the progress made in ACTF Flow Loop development and research during 90 days pre-award period (May 15-July 14, 1999) and the following three months after the project approval date (July15-October 15, 1999) The report presents information on the following specific subjects; (a) Progress in Advanced Cuttings Transport Facility design and development, (b) Progress report on the research project ''Study of Flow of Synthetic Drilling Fluids Under Elevated Pressure and Temperature Conditions'', (c) Progress report on the research project ''Study of Cuttings Transport with Foam Under LPAT Conditions (Joint Project with TUDRP)'', (d) Progress report on the research project ''Study of Cuttings Transport with Aerated Muds Under LPAT Conditions (Joint Project with TUDRP)'', (e) Progress report on the research project ''Study of Foam Flow Behavior Under EPET Conditions'', (f) Progress report on the instrumentation tasks (Tasks 11 and 12) (g) Activities towards technology transfer and developing contacts with oil and service company members.

  5. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Troy Reed; Stefan Miska; Nicholas Takach; Kaveh Ashenayi; Mark Pickell; Len Volk; Mike Volk; Lei Zhou; Zhu Chen; Crystal Redden; Aimee Washington

    2003-07-30

    This Quarter has been divided between running experiments and the installation of the drill-pipe rotation system. In addition, valves and piping were relocated, and three viewports were installed. Detailed design work is proceeding on a system to elevate the drill-string section. Design of the first prototype version of a Foam Generator has been finalized, and fabrication is underway. This will be used to determine the relationship between surface roughness and ''slip'' of foams at solid boundaries. Additional cups and rotors are being machined with different surface roughness. Some experiments on cuttings transport with aerated fluids have been conducted at EPET. Theoretical modeling of cuttings transport with aerated fluids is proceeding. The development of theoretical models to predict frictional pressure losses of flowing foam is in progress. The new board design for instrumentation to measure cuttings concentration is now functioning with an acceptable noise level. The ultrasonic sensors are stable up to 190 F. Static tests with sand in an annulus indicate that the system is able to distinguish between different sand concentrations. Viscometer tests with foam, generated by the Dynamic Test Facility (DTF), are continuing.

  6. High-Throughput Plasmid cDNA Library Screening

    Energy Technology Data Exchange (ETDEWEB)

    Wan, Kenneth H.; Yu, Charles; George, Reed A.; Carlson, JosephW.; Hoskins, Roger A.; Svirskas, Robert; Stapleton, Mark; Celniker, SusanE.

    2006-05-24

    Libraries of cDNA clones are valuable resources foranalysing the expression, structure, and regulation of genes, as well asfor studying protein functions and interactions. Full-length cDNA clonesprovide information about intron and exon structures, splice junctionsand 5'- and 3'-untranslated regions (UTRs). Open reading frames (ORFs)derived from cDNA clones can be used to generate constructs allowingexpression of native proteins and N- or C-terminally tagged proteins.Thus, obtaining full-length cDNA clones and sequences for most or allgenes in an organism is critical for understanding genome functions.Expressed sequence tag (EST) sequencing samples cDNA libraries at random,which is most useful at the beginning of large-scale screening projects.However, as projects progress towards completion, the probability ofidentifying unique cDNAs via EST sequencing diminishes, resulting in poorrecovery of rare transcripts. We describe an adapted, high-throughputprotocol intended for recovery of specific, full-length clones fromplasmid cDNA libraries in five days.

  7. Performance Testing of Cutting Fluids

    DEFF Research Database (Denmark)

    Belluco, Walter

    within the whole range of operations, materials, cutting fluids, operating conditions, etc. Cutting fluid performance was evaluated in turning, drilling, reaming and tapping, and with respect to tool life, cutting forces, chip formation and product quality (dimensional accuracy and surface integrity......). A number of different work materials were considered, with emphasis on austenitic stainless steel. Cutting fluids from two main groups were investigated, water miscible (reviewed from previous work) and straight oils. Results show that correlation of cutting fluid performance in different operations exists...... within the same group of cutting fluids, for stainless steel. A possible rationalisation of cutting fluid performance tests is suggested. In order to select a set of basic tests and optimise them for use as general and standardised testing methods, an original approach to the evaluation of cutting force...

  8. Performance Testing of Cutting Fluids

    DEFF Research Database (Denmark)

    Belluco, Walter

    The importance of cutting fluid performance testing has increased with documentation requirements of new cutting fluid formulations based on more sustainable products, as well as cutting with minimum quantity of lubrication and dry cutting. Two sub-problems have to be solved: i) which machining...... tests feature repeatability, reproducibility and sensitivity to cutting fluids, and ii) to what extent results of one test ensure relevance to a wider set of machining situations. The present work is aimed at assessing the range of validity of the different testing methods, investigating correlation...... within the whole range of operations, materials, cutting fluids, operating conditions, etc. Cutting fluid performance was evaluated in turning, drilling, reaming and tapping, and with respect to tool life, cutting forces, chip formation and product quality (dimensional accuracy and surface integrity...

  9. Cutting the Cord-2

    Science.gov (United States)

    2004-01-01

    This animation shows the view from the rear hazard avoidance cameras on the Mars Exploration Rover Spirit as the rover turns 45 degrees clockwise. This maneuver is the first step in a 3-point turn that will rotate the rover 115 degrees to face west. The rover must make this turn before rolling off the lander because airbags are blocking it from exiting from the front lander petal. Before this crucial turn took place, engineers instructed the rover to cut the final cord linking it to the lander. The turn took around 30 minutes to complete.

  10. Cutting the Cord

    Science.gov (United States)

    2004-01-01

    This animation shows the view from the front hazard avoidance cameras on the Mars Exploration Rover Spirit as the rover turns 45 degrees clockwise. This maneuver is the first step in a 3-point turn that will rotate the rover 115 degrees to face west. The rover must make this turn before rolling off the lander because airbags are blocking it from exiting off the front lander petal. Before this crucial turn could take place, engineers instructed the rover to cut the final cord linking it to the lander. The turn took around 30 minutes to complete.

  11. Drilling cost-cutting

    Energy Technology Data Exchange (ETDEWEB)

    Capuano, L.E. Jr.

    1996-12-31

    This presentation by Louis E. Capuano, Jr., President, ThermaSource, Inc., discusses cost-cutting in the drilling phase of geothermal energy exploration and production. All aspects of a geothermal project including the drilling must be streamlined to make it viable and commercial. If production could be maximized from each well, there would be a reduction in drilling costs. This could be achieved in several ways, including big hole and multi-hole completion, directional drilling, better knowledge of the resource and where to penetrate, etc.

  12. Cloning and expression analysis of human reticulon 4c cDNA

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    RTNs (reticulons) is a gene family related to the growth and differentiation of neuroendocrine cell. This family is composed of several members such as RTN1, RTN2 and RTN3. RTN1 and RTN2 have been proved to have 3 transcripts with different length. Because the RTN1c cDNA was involved in the sologenesis of small cell lung carcinoma (SCLC), it was selected as a bioinformatic probe to clone novel members of RTN family with the electric hybridization assistant new-gene cloning method (EHAC). A 1677-bp cDNA was identified from human brain cDNA library. The cDNA contains an intact open reading frame (ORF) which encodes a protein of 199 amino acids. This deduced protein is highly homologous to RTN1c, RTN2c and RTN3 with identities of 64.4%, 45.8% and 50.0% respectively. This new gene was named RTN4c (GenBank accession number: AF087901). Northern hybridization showed that the full length of RTN4c transcript is about 1.8 kb. It is hardly expressed in heart, placenta, lung, spleen, thymus, testis, ovary, small intestine and peripheral blood white cells; but it is highly expressed in the tissues of skeletal muscle, brain, liver and kidney, and less expressed in the pancreas, prostate and colon. Furthermore, Northern results also showed that there is a 2.3 kb transcript expressed in 14 tissues except liver and skeletal muscle; while another 5.0 kb transcript in brain, skeletal muscle and testis. By the electric hybridization walking, we obtained two full-length contigs with a length of 4632 and 2235 bp respectively. The former encodes a protein with 1192 amino acids and was defined as RTN4a; the latter encodes another protein with 373 amino acids, and was named RTN4b. The RTN4 gene was mapped to human chromosome 2p14-p13 region by the radiation hybridization (RH).

  13. Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate

    Directory of Open Access Journals (Sweden)

    Murwantoko M

    2015-11-01

    Full Text Available Rhoptry protein belongs to an excretory and secretory antigens (ESAs that play an important role during active penetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targeted cell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasite successfully enter the cell target then Granule (GRA proteins are responsible for the formation of parasitophorus vacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently, this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone and sequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique. Total ribonucleic acid (RNA was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA was used as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor from Riboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinant plasmid was transformed into E. coli (XL1-Blue. The transformed E. coli XL-1 Blue were plated on LB agar containing X-Gal, IPTG and ampicillin. Recombinant clones (white colony were picked up and grown up in the LB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order to identify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolated using alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid was cut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward and M13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretory

  14. Improved performance of linear coal cutting compared with rotary cutting

    Energy Technology Data Exchange (ETDEWEB)

    Roepke, W.W.; Hanson, B.D.; Olson, R.C.; Wingquist, C.F.; Myren, T.A.

    1995-09-01

    The linear cutting system, developed by the US Bureau of Mines uses geometric principles developed by Cardan to produce a nearly constant cut depth. The new system has been extensively tested in a synthetic material under laboratory conditions to verify mechanical capability and to identify operational characteristics. Comparison between 15-rpm linear cutting and 50-rpm rotary cutting systems show significant improvement in respirable dust entrainment, product size distribution, and energy usage. Respirable dust is reduced by as much as 90%. Recovered product showed a 67% reduction in {minus}0.32-cm ({minus}1/8-in) material and a 200% increase in +5.08 cm (+ 2 in) materials. Average power was reduced by 66% for the linear cutting. Because the bit cutting paths differ between linear and rotary cutting, it was necessary to compare the two at the same cut depths and bit types. These comparisons show that low revolution per minute rotary cutting entrains about the same amount of respirable dust as the linear cutting system, but the average shaft torque may be 55 to 130% greater for the rotary system.

  15. CO2 laser cutting

    CERN Document Server

    Powell, John

    1998-01-01

    The laser has given manufacturing industry a new tool. When the laser beam is focused it can generate one of the world's most intense energy sources, more intense than flames and arcs, though similar to an electron beam. In fact the intensity is such that it can vaporise most known materials. The laser material processing industry has been growing swiftly as the quality, speed and new manufacturing possibilities become better understood. In the fore of these new technologies is the process of laser cutting. Laser cutting leads because it is a direct process substitu­ tion and the laser can usually do the job with greater flexibility, speed and quality than its competitors. However, to achieve these high speeds with high quality con­ siderable know how and experience is required. This information is usually carefully guarded by the businesses concerned and has to be gained by hard experience and technical understanding. Yet in this book John Powell explains in lucid and almost non­ technical language many o...

  16. Preparation of cDNA libraries from vascular cells.

    Science.gov (United States)

    Lieb, M E; Taubman, M B

    1999-01-01

    The vast majority of past and present efforts in the molecular cloning of expressed sequences involve isolation of clones from cDNA libraries constructed in bacteriophage lambda (1,2). As discussed in Chapter 6 , screening these cDNA libraries using labeled probes remains the most straightforward method to isolate full length cDNAs for which some partial sequence information is known. Although the availability of high quality reagents and kits over the past decade has made the process of library construction increasingly straightforward, generation of high-quality libraries is a task that still requires a fair amount of dedicated effort. Because alternative PCR-based cloning strategies have become increasingly popular alternatives to cDNA library screening, it is useful to consider the advantages and disadvantages of each strategy before embarking on a project to construct a cDNA library (Table 1). In our opinion, it is worthwhile to construct a cDNA library when the transcript of interest is not exceedingly rare (i.e., can readily be detected by Northern blot analysis of total RNA), when multiple cDNAs will need to be cloned over a period of time, and in situations where occasional mutations can not be tolerated (for example, if the cDNA is to be expressed in mammalian cells to examine function). In situations where the transcript of interest is expressed at exceedingly low levels, or when only a single cDNA needs to be cloned, a PCR-based strategy should be considered. When the tissue source is precious (such as a unique clinical specimen), successful construction of a phage library provides a resource that can be amplified and used for multiple cloning projects over many years, but runs the risk of consuming the available RNA if the library construction fails. Table 1 Comparison of Relative Advantages of cDNA Cloning from Lambda Phage Libraries by Plaque Hybridization Compared to Newer PCR- Based Strategies Lambda phage cDNA library PCR-based strategy Freedom

  17. Discovery of Phytophthora infestans genes expressed in planta through mining of cDNA libraries.

    Directory of Open Access Journals (Sweden)

    Roberto Sierra

    Full Text Available BACKGROUND: Phytophthora infestans (Mont. de Bary causes late blight of potato and tomato, and has a broad host range within the Solanaceae family. Most studies of the Phytophthora--Solanum pathosystem have focused on gene expression in the host and have not analyzed pathogen gene expression in planta. METHODOLOGY/PRINCIPAL FINDINGS: We describe in detail an in silico approach to mine ESTs from inoculated host plants deposited in a database in order to identify particular pathogen sequences associated with disease. We identified candidate effector genes through mining of 22,795 ESTs corresponding to P. infestans cDNA libraries in compatible and incompatible interactions with hosts from the Solanaceae family. CONCLUSIONS/SIGNIFICANCE: We annotated genes of P. infestans expressed in planta associated with late blight using different approaches and assigned putative functions to 373 out of the 501 sequences found in the P. infestans genome draft, including putative secreted proteins, domains associated with pathogenicity and poorly characterized proteins ideal for further experimental studies. Our study provides a methodology for analyzing cDNA libraries and provides an understanding of the plant--oomycete pathosystems that is independent of the host, condition, or type of sample by identifying genes of the pathogen expressed in planta.

  18. cDNA cloning, characterization, and expression analysis of the Rac1 gene from Scophthalmus maximus.

    Science.gov (United States)

    Jia, Airong; Zhang, Xiao-Hua

    2009-09-01

    Rac1 is a small GTP-binding protein that belongs to the Rho small GTPases, which are important signaling molecules that regulate the dynamics of the actin cytoskeleton and mediate changes in cell morphology and motility. The EST sequence of Rac1 from turbot (Scophthalmus maximus L.) was obtained from a subtractive cDNA library previously. In this study, the full-length cDNA sequence of turbot Rac1 was obtained, which was 2420 nucleotides (nt) encoding a protein of 192 amino acids, with a putative molecular weight of 21.3 kDa. At the amino-acid level, turbot Rac1 was highly conserved to previously characterized GTPases of Rac sub-family, and was nearly identical to human Rac1 (95.3% identity). Quantitative real-time PCR demonstrated that the Rac1 was constitutively expressed in all tissues examined, but at different levels. Upon challenge with Vibrio harveyi, the expression level of Rac1 fluctuated in the liver at different time points. In the head kidney, its expression level decreased to the lowest at 4 h, and then increased to the background level at 24 h. The remarkable degree of evolutionary conservation observed in turbot Rac1 primary structure together with its changing in expression level upon challenge suggested a functionally important role for this Rho family member in the immune response.

  19. cDNA cloning and expression of earthworm fibrinolytic enzyme component A

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Earthworm fibrinolytic enzyme component A (EFEa), a protein with dual fibrinolytic activity, is one of the major therapeutically important earthworm fibrinolytic enzyme components. The cDNA fragment encoded the mature protein was cloned from earthworm (Eisenia fetida) by the RT-PCR technique. The deduced amino acid sequence of the EFE component A shows high homology with some members of serine proteases trypsin family, and the amino acid residues constituting the active sites are conserved in the EFEa as compared with the other proteins of the trypsin family. The cDNA fragment was subcloned into the expression vector pQE31 and pMAL-c2X of E. coli. The resulting expression plasmids, pQE-efea and pMAL-efea, were used to transform the E. coli strain M15. Recombinant protein bands corresponding with calculated molecular weights were induced. The induced His6-EFEa fusion protein with pQE- efea was accumulated into inclusion body, while the induced MBP-EFEa fusion protein with pMAL-efea was soluble and showed fibrinolytic activities.

  20. Rapid amplification of cDNA ends (RACE).

    Science.gov (United States)

    Yeku, Oladapo; Frohman, Michael A

    2011-01-01

    Rapid Amplification of cDNA ends (RACE) provides an inexpensive and powerful tool to quickly obtain full-length cDNA when the sequence is only partially known. Starting with an mRNA mixture, gene-specific primers generated from the known regions of the gene and non-specific anchors, full-length sequences can be identified in as little as 3 days. RACE can also be used to identify alternative transcripts of a gene when the partial or complete sequence of only one transcript is known. In the following sections, we outline details for rapid amplification of 5(') and 3(') cDNA ends using the "new RACE" technique.

  1. Procedure for normalization of cDNA libraries

    Science.gov (United States)

    Bonaldo, Maria DeFatima; Soares, Marcelo Bento

    1997-01-01

    This invention provides a method to normalize a cDNA library constructed in a vector capable of being converted to single-stranded circles and capable of producing complementary nucleic acid molecules to the single-stranded circles comprising: (a) converting the cDNA library in single-stranded circles; (b) generating complementary nucleic acid molecules to the single-stranded circles; (c) hybridizing the single-stranded circles converted in step (a) with complementary nucleic acid molecules of step (b) to produce partial duplexes to an appropriate Cot; (e) separating the unhybridized single-stranded circles from the hybridized single-stranded circles, thereby generating a normalized cDNA library.

  2. Cloning and expression analysis of MBLL cDNA

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The mbl (muscleblind) gene of Drosophila encodes a nuclear protein which contains two Cys3His motifs. The mutation of mbl gene will disturb the differentiation of all the Drosophila's photoreceptors. Primers have been designed according to human EST086139, which is highly homologous to mbl gene. Human fetal brain cDNA library has been screened and a novel cDNA clone has been obtained. The 2595 bp cDNA, designated MBLL (muscleblind-like), contains an open reading frame which encodes 255 amino acids and has 4 Cys3His motifs (GenBank Acc. AF061261). The amino acids sequence shares high homology to Drosophila's mbl. The Northern blot and RNA dot blot hybridization of 43 human adult tissues and 7 fetal tissues show that MBLL is a widely expressed gene, but the expression amounts differ in these tissues.

  3. ADVANCED CUTTINGS TRANSPORT STUDY

    Energy Technology Data Exchange (ETDEWEB)

    Stefan Miska; Nicholas Takach; Kaveh Ashenayi

    2004-01-31

    Final design of the mast was completed (Task 5). The mast is consisting of two welded plate girders, set next to each other, and spaced 14-inches apart. Fabrication of the boom will be completed in two parts solely for ease of transportation. The end pivot connection will be made through a single 2-inch diameter x 4 feet-8 inch long 316 SS bar. During installation, hard piping make-ups using Chiksan joints will connect the annular section and 4-inch return line to allow full movement of the mast from horizontal to vertical. Additionally, flexible hoses and piping will be installed to isolate both towers from piping loads and allow recycling operations respectively. Calibration of the prototype Foam Generator Cell has been completed and experiments are now being conducted. We were able to generate up to 95% quality foam. Work is currently underway to attach the Thermo-Haake RS300 viscometer and install a view port with a microscope to measure foam bubble size and bubble size distribution. Foam rheology tests (Task 13) were carried out to evaluate the rheological properties of the proposed foam formulation. After successful completion of the first foam test, two sets of rheological tests were conducted at different foam flow rates while keeping other parameters constant (100 psig, 70F, 80% quality). The results from these tests are generally in agreement with the previous foam tests done previously during Task 9. However, an unanticipated observation during these tests was that in both cases, the frictional pressure drop in 2 inch pipe was lower than that in the 3 inch and 4 inch pipes. We also conducted the first foam cuttings transport test during this quarter. Experiments on aerated fluids without cuttings have been completed in ACTF (Task 10). Gas and liquid were injected at different flow rates. Two different sets of experiments were carried out, where the only difference was the temperature. Another set of tests was performed, which covered a wide range of

  4. Vortex cutting in superconductors

    Science.gov (United States)

    Glatz, A.; Vlasko-Vlasov, V. K.; Kwok, W. K.; Crabtree, G. W.

    2016-08-01

    Vortex cutting and reconnection is an intriguing and still-unsolved problem central to many areas of classical and quantum physics, including hydrodynamics, astrophysics, and superconductivity. Here, we describe a comprehensive investigation of the crossing of magnetic vortices in superconductors using time dependent Ginsburg-Landau modeling. Within a macroscopic volume, we simulate initial magnetization of an anisotropic high temperature superconductor followed by subsequent remagnetization with perpendicular magnetic fields, creating the crossing of the initial and newly generated vortices. The time resolved evolution of vortex lines as they approach each other, contort, locally conjoin, and detach, elucidates the fine details of the vortex-crossing scenario under practical situations with many interacting vortices in the presence of weak pinning. Our simulations also reveal left-handed helical vortex instabilities that accompany the remagnetization process and participate in the vortex crossing events.

  5. Laser Cutting of Different Materials

    Directory of Open Access Journals (Sweden)

    Kadir ÇAVDAR

    2013-08-01

    Full Text Available In this paper; in general potential developments and trends of a particular machining field by extensively evaluating present studies of laser beam machining have been discussed. As it is indicated below, technical literatures have been subsumed under five major headlines: Experimental studies, reviews, optimization researches of the cutting parameters, theoretical modelling studies of laser beam cutting and academic studies relating to laser cutting

  6. Cut and Column Generation

    DEFF Research Database (Denmark)

    Spoorendonk, Simon

    artikel, men det vises, at det er muligt at finde særdeles gode grænseværdier ved tilføjelsen af denne familie af snit. I den tredje artikel betragtes klike (eng.: clique) uligheder, som er et kombinatorisk snit til master problemet for ruteplanlægningsproblemet med tidsvinduer. I tråd med arbejdet i de...

  7. CALCULATION OF LASER CUTTING COSTS

    Directory of Open Access Journals (Sweden)

    Bogdan Nedic

    2016-09-01

    Full Text Available The paper presents description methods of metal cutting and calculation of treatment costs based on model that is developed on Faculty of mechanical engineering in Kragujevac. Based on systematization and analysis of large number of calculation models of cutting with unconventional methods, mathematical model is derived, which is used for creating a software for calculation costs of metal cutting. Software solution enables resolving the problem of calculating the cost of laser cutting, comparison' of costs made by other unconventional methods and provides documentation that consists of reports on estimated costs.

  8. A BIOINFORMATIC STRATEGY TO RAPIDLY CHARACTERIZE CDNA LIBRARIES

    Science.gov (United States)

    A Bioinformatic Strategy to Rapidly Characterize cDNA LibrariesG. Charles Ostermeier1, David J. Dix2 and Stephen A. Krawetz1.1Departments of Obstetrics and Gynecology, Center for Molecular Medicine and Genetics, & Institute for Scientific Computing, Wayne State Univer...

  9. NORMAL NASAL GENE EXPRESSION LEVELS USING CDNA ARRAY TECHNOLOGY

    Science.gov (United States)

    Normal Nasal Gene Expression Levels Using cDNA Array Technology. The nasal epithelium is a target site for chemically-induced toxicity and carcinogenicity. To detect and analyze genetic events which contribute to nasal tumor development, we first defined the gene expressi...

  10. Rescue of mumps virus from cDNA.

    Science.gov (United States)

    Clarke, D K; Sidhu, M S; Johnson, J E; Udem, S A

    2000-05-01

    A complete DNA copy of the genome of a Jeryl Lynn strain of mumps virus (15,384 nucleotides) was assembled from cDNA fragments such that an exact antigenome RNA could be generated following transcription by T7 RNA polymerase and cleavage by hepatitis delta virus ribozyme. The plasmid containing the genome sequence, together with support plasmids which express mumps virus NP, P, and L proteins under control of the T7 RNA polymerase promoter, were transfected into A549 cells previously infected with recombinant vaccinia virus (MVA-T7) that expressed T7 RNA polymerase. Rescue of infectious virus from the genome cDNA was demonstrated by amplification of mumps virus from transfected-cell cultures and by subsequent consensus sequencing of reverse transcription-PCR products generated from infected-cell RNA to verify the presence of specific nucleotide tags introduced into the genome cDNA clone. The only coding change (position 8502, A to G) in the cDNA clone relative to the consensus sequence of the Jeryl Lynn plaque isolate from which it was derived, resulting in a lysine-to-arginine substitution at amino acid 22 of the L protein, did not prevent rescue of mumps virus, even though an amino acid alignment for the L proteins of paramyxoviruses indicates that lysine is highly conserved at that position. This system may provide the basis of a safe and effective virus vector for the in vivo expression of immunologically and biologically active proteins, peptides, and RNAs.

  11. Dominus for cut flower production

    Science.gov (United States)

    Fumigation with methyl bromide was the principal method of soilborne pest control in cut flower production. Many cut flower growers in Florida have ceased production, but those that remain are restricted in the fumigants that they are able to utilize due to proximity to potable water sources and oc...

  12. Will Intel Grow Through Cuts?

    Institute of Scientific and Technical Information of China (English)

    DAN; STEINBOCK

    2006-01-01

    Short-term cuts, perhaps, but long-term growth in China may be inevitable T he world's largest chipmaker, Intel, is sending a ripple effect through its pool of labor globally, including in China, with the announcement of deep workforce cuts.

  13. Rimbaud Cut-Up Burroughs

    Directory of Open Access Journals (Sweden)

    Mauricio Vasconcelos

    1996-10-01

    Full Text Available O objetivo deste ensaio é o de explorar a relação entre a poesia de Rimbaud e as experiências de William Burroughs com os cut-ups. A influência de Rimbaud sobre o trabalho de Burroughs se estende, contudo, para além dos cut-ups, envolvendo sua escrita ficcional.

  14. Ritual genital cutting of female minors.

    Science.gov (United States)

    Davis, Dena S

    2010-05-01

    The traditional custom of ritual cutting and alteration of the genitalia of female infants, children, and adolescents, referred to as female genital mutilation or female genital cutting (FGC), persists primarily in Africa and among certain communities in the Middle East and Asia. Immigrants in the United States from areas in which FGC is common may have daughters who have undergone a ritual genital procedure or may request that such a procedure be performed by a physician. The American Academy of Pediatrics believes that pediatricians and pediatric surgical specialists should be aware that this practice has life-threatening health risks for children and women. The American Academy of Pediatrics opposes all types of female genital cutting that pose risks of physical or psychological harm, counsels its members not to perform such procedures, recommends that its members actively seek to dissuade families from carrying out harmful forms of FGC, and urges its members to provide patients and their parents with compassionate education about the harms of FGC while remaining sensitive to the cultural and religious reasons that motivate parents to seek this procedure for their daughters.

  15. OPTIMAL CONTROL OF CNC CUTTING PROCESS

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    The intelligent optimizing method of cutting parameters and the cutting stable districts searching method are set up. The cutting parameters of each cutting pass could be optimized automatically, the cutting chatter is predicted through setting up the dynamic cutting force AR(2) model on-line, the spindle rotation speed is adjusted according to the predicting results so as to ensure the cutting system work in stable district.

  16. Online Cake Cutting (published version)

    CERN Document Server

    Walsh, Toby

    2011-01-01

    We propose an online form of the cake cutting problem. This models situations where agents arrive and depart during the process of dividing a resource. We show that well known fair division procedures like cut-and-choose and the Dubins-Spanier moving knife procedure can be adapted to apply to such online problems. We propose some fairness properties that online cake cutting procedures can possess like online forms of proportionality and envy-freeness. We also consider the impact of collusion between agents. Finally, we study theoretically and empirically the competitive ratio of these online cake cutting procedures. Based on its resistance to collusion, and its good performance in practice, our results favour the online version of the cut-and-choose procedure over the online version of the moving knife procedure.

  17. Molecular Cloning of Myostatin Partial cDNA of Beijing Duck and Its Expression in Breast Muscle

    Institute of Scientific and Technical Information of China (English)

    WANG Yong-sheng; HOU Shui-sheng; HUANG Wei; KANG Jun-mei

    2006-01-01

    In this experiment, 500 bp cDNA of myostatin gene was cloned from a Beijing duck's breast. The duck myostatin gene was found to have 98, 96, 95, 88, and 87% sequence similarity at the cDNA level with domestic goose, chicken, domestic pigeon, human, and pig, respectively. The predicted amino acid sequence has an overall similarity with a comparable region of turkey 99%, domestic goose 98%, and chicken 99%. Conserved domains of deduced amino acids showed that it belonged to the TGF-beta family. Myostatin expression in breast muscle was higher at 28, 35, and 42 days than at 7, 14, and 21 days. The pattern of myostatin expression was closely parallel to the trend of breast muscle growth, suggesting that myostatin might play an important role in breast muscle development. It was possible to postulate that myostatin may be a major determinant of muscle mass in breast muscle, as shown in other species.

  18. Cutting forces in orthogonal cutting of unidirectional GFRP composites

    Energy Technology Data Exchange (ETDEWEB)

    Caprino, G.; Nele, L. [Univ. of Naples Federico II (Italy). Dept. of Materials and Production Engineering

    1996-07-01

    The results of orthogonal cutting tests carried out on unidirectional glass fiber reinforced plastic composites, using HSS tools, are presented and discussed. During the tests, performed on a milling machine at very low cutting speed to avoid thermal effects, the cutting speed was held constant and parallel to the fiber direction. Three parameters, namely the tool rake angle {alpha}, the tool relief angle {gamma}, and the depth of cut t, were varied. According to the experimental results, the horizontal force per unit width, F{sub hu}, undergoes a dramatic decrease, never verified for metals, with increasing {alpha}. Besides, F{sub hu} is only negligibly affected by the relief angle, and linearly increases with t. Similarly to metals, an effect of the depth of cut on the specific energy (size effect) is found also for composites. However, the presented results indicate that the size effect can be analytically modeled in a simple way in the case of composites. The vertical force per unit width, F{sub vu}, exhibits a marked reduction when the relief angle is increased. F{sub vu} is also very sensitive to the rake angle: the lower {alpha}, the higher is F{sub vu}. It is shown that this behavior probably reflects a strong influence of the rake angle on the forces developing at the flank. A linear dependence of the vertical force on the depth of cut is also demonstrated. Finally, the experimental data are utilized to obtain empirical formulae, allowing an approximate evaluation of cutting forces.

  19. Construction and biological activities of the first infectious cDNA clones of the genus Foveavirus

    Energy Technology Data Exchange (ETDEWEB)

    Meng, Baozhong, E-mail: bmeng@uoguelph.ca [Department of Molecular and Cellular Biology, University of Guelph, 50 Stone Road, Guelph, Ontario, Canada N1G2W1 (Canada); Venkataraman, Srividhya; Li, Caihong; Wang, Weizhou [Department of Molecular and Cellular Biology, University of Guelph, 50 Stone Road, Guelph, Ontario, Canada N1G2W1 (Canada); Dayan-Glick, Cathy; Mawassi, Munir [The Plant Pathology Department-The Virology Unit, Plant Protection Institute, Agricultural Research Organization, The Volcani Center, Bet-Dagan 50250 (Israel)

    2013-01-20

    Grapevine rupestris stem pitting-associated virus (GRSPaV, genus Foveavirus, family Betaflexiviridae) is one of the most prevalent viruses in grapevines and is associated with three distinct diseases: rupestris stem pitting, vein necrosis and Syrah decline. Little is known about the biology and pathological properties of GRSPaV. In this work, we engineered a full-length infectious cDNA clone for GRSPaV and a GFP-tagged variant, both under the transcriptional control of Cauliflower mosaic virus 35 S promoter. We demonstrated that these cDNA clones were infectious in grapevines and Nicotiana benthamiana through fluorescence microscopy, RT-PCR, Western blotting and immuno electron microscopy. Interestingly, GRSPaV does not cause systemic infection in four of the most commonly used herbaceous plants, even in the presence of the movement proteins of two other viruses which are known to complement numerous movement-defective viruses. These infectious clones are the first of members of Foveavirus which would allow further investigations into mechanisms governing different aspects of replication for GRSPaV and perhaps related viruses.

  20. Nucleotide sequence of cloned cDNA for human pancreatic kallikrein.

    Science.gov (United States)

    Fukushima, D; Kitamura, N; Nakanishi, S

    1985-12-31

    Cloned cDNA sequences for human pancreatic kallikrein have been isolated and determined by molecular cloning and sequence analysis. The identity between human pancreatic and urinary kallikreins is indicated by the complete coincidence between the amino acid sequence deduced from the cloned cDNA sequence and that reported partially for urinary kallikrein. The active enzyme form of the human pancreatic kallikrein consists of 238 amino acids and is preceded by a signal peptide and a profragment of 24 amino acids. A sequence comparison of this with other mammalian kallikreins indicates that key amino acid residues required for both serine protease activity and kallikrein-like cleavage specificity are retained in the human sequence, and residues corresponding to some external loops of the kallikrein diverge from other kallikreins. Analyses by RNA blot hybridization, primer extension, and S1 nuclease mapping indicate that the pancreatic kallikrein mRNA is also expressed in the kidney and sublingual gland, suggesting the active synthesis of urinary kallikrein in these tissues. Furthermore, the tissue-specific regulation of the expression of the members of the human kallikrein gene family has been discussed.

  1. RNA transcripts of full-length cDNA clones of rabbit hepatitis E virus are infectious in rabbits.

    Science.gov (United States)

    Cossaboom, Caitlin M; Huang, Yao-Wei; Yugo, Danielle M; Kenney, Scott P; Piñeyro, Pablo; Matzinger, Shannon R; Heffron, C Lynn; Pierson, F William; Meng, Xiang-Jin

    2014-11-07

    Hepatitis E virus (HEV), the causative agent of hepatitis E, is a single-stranded positive-sense RNA virus belonging to the family Hepeviridae. At least four genotypes of the family infect humans: genotypes 1 and 2 are transmitted to humans through contaminated water, while genotypes 3 and 4 are zoonotic and have animal reservoirs. A novel strain of HEV recently identified in rabbits is a distant member of genotype 3, and thus poses a potential risk of zoonotic transmission to humans. The objective of this study was to construct and characterize an infectious cDNA clone of the rabbit HEV. Two full-length cDNA clones of rabbit HEV, pT7g-rabHEV and pT7-rabHEV, were constructed and their infectivity was tested by in vitro transfection of Huh7 human liver cells and by direct intrahepatic inoculation of rabbits with capped RNA transcripts. Results showed that positive signal for rabbit HEV protein was detected by an immunofluorescence assay with a HEV-specific antibody in Huh7 human liver cells transfected with capped RNA transcripts from the two full-length cDNA clones. Rabbits intrahepatically inoculated with capped RNA transcripts from each of the two clones developed active HEV infection as evidenced by seroconversion to anti-HEV antibodies, and detection of rabbit HEV RNA in sera and feces of inoculated animals. The availability of a rabbit HEV infectious cDNA clone now affords us the ability to delineate the mechanism of HEV replication and cross-species infection in a small animal model.

  2. "Paper Cut Lady" Ku Shulan

    Institute of Scientific and Technical Information of China (English)

    1995-01-01

    WHEN the people in Xunyi County of Shaanxi Province talk about the traditional paper cut, they always speak of a "paper cut lady" Ku Shulan. Her works have been shown at the Xi’an Artists’ Gallery, Beijing China Art Gallery and Exhibition Hall of Central Academy of Fine Arts. Almost seventy years old, Ku Shulan is an ordinary country lady, and her artistic achievements were closely linked with her mother countryside. The traditional paper cut created and developed in Xunyi County, has a simple and bold style of Northern Shaanxi, and an exquisitely

  3. Detection of germline mutations of hMLH1 and hMSH2 based on cDNA sequencing in China

    Institute of Scientific and Technical Information of China (English)

    Chao-Fu Wang; Xiao-Yan Zhou; Tai-Ming Zhang; Meng-Hong Sun; Da-Ren Shi

    2005-01-01

    AIM: To detect the germline mutations of hMLH1 and hMSH2based on mRNA sequencing to identify hereditary nonpolyposis colorectal cancer (HNPCC) families.METHODS: Total RNA was extracted from peripheral blood of 14 members from 12 different families fulfilling Amsterdam criteria Ⅱ. mRNA of hMLH1 and hMSH2 was reversed with special primers and heat-resistant reverse transcriptase. cDNA was amplified with expand long template PCR and cDNA sequendng analysis was followed.RESULT: Seven germline mutations were found in 6families (6/12, 50%), in 4 hMLH1 and 3 hMSH2 mutations (4/12, 33.3%); (3/12, 25%). The mutation types involved 4 missense, 1 silent and 1 frame shift mutations as well as 1 mutation in the non-coding area. Four out of the seven mutations have not been reported previously. The 4 hMLH1mutations were distributed in exons 8, 12, 16, and 19. The 3hMSH2 mutations were distributed in exons 1 and 2. Six out of the 7 mutations were pathological, which were distributed in 5 HNPCC families.CONCLUSION: Germline mutations of hMLH1 and hMSH2 can be found based on cDNA sequencing so as to identify HNPCC family, which is highly sensitive and has the advantages of cost and time saving.

  4. cDNA cloning, chromosome mapping and expression characterization of human geranylgeranyl pyrophosphate synthase

    Institute of Scientific and Technical Information of China (English)

    赵勇[1; 余龙[2; 高洁[3; 付强[4; 华益民[5; 张宏来[6; 赵寿元[7

    2000-01-01

    Geranylgeranyl pyrophosphate (GGPP) mainly participates in post-translational modification for various proteins including Rho/Rac, Rap and Rab families, as well as in regulation for cell apoptosis. Geranylgeranyl pyrophosphate synthase (GGPPS), which catalyzes the condensation reaction between farnesyl diphosphate and isopentenyl diphosphate, is the key enzyme for synthesizing GGPP. We report the isolation of a gene transcript showing high homology with Drosophila GGPPS cDNA. The transcript is 1 466 bp in length and contains an intact open reading frame (ORF) ranging from nt 239 to 1 138. This ORF encodes a deduced protein of 300 residues with calculated molecular weight of 35 ku. The deduced protein shows 57.5% identity and 75% similarity with Drosophila GGPPS, and contains five characteristic domains of prenyltransferases. Northern hybridization revealed that human GGPPS was expressed highest in heart, and moderately in spleen, testis, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas

  5. AHM cutting expenses, adding technology.

    Science.gov (United States)

    Lutz, S

    1990-04-30

    American Healthcare Management, emerging from bankruptcy protection is cutting corporate costs and expanding into new technology to attract more private-paying patients and lessen dependence on Medicare business.

  6. Refrigerated cutting tools improve machining of superalloys

    Science.gov (United States)

    Dudley, G. M.

    1971-01-01

    Freon-12 applied to tool cutting edge evaporates quickly, leaves no residue, and permits higher cutting rate than with conventional coolants. This technique increases cutting rate on Rene-41 threefold and improves finish of machined surface.

  7. CDNA encoding a polypeptide including a hevein sequence

    Science.gov (United States)

    Raikhel, Natasha V.; Broekaert, Willem F.; Chua, Nam-Hai; Kush, Anil

    1995-03-21

    A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.

  8. cDNA table - RPD | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us RPD cDNA table Data detail Data name cDNA table DOI 10.18908/lsdba.nbdc00191-004 Description...age About This Database Database Description Download License Update History of This Database Site Policy | Contact Us cDNA table - RPD | LSDB Archive ...

  9. Optimization of Cutting Variables in Machining

    Institute of Scientific and Technical Information of China (English)

    Zhang Xueyan; Wu Zhenye; Zhou Guohua

    1996-01-01

    With the three criteria for cutting variables proposed by W. W. Gilbert and K.Hitomi, th is paper analyzes the reasonable selection of cutting variables, and further states the relations among maximum profit-oriented cutting speed,minimum cost-oriented cutting speed and maximum productivity-oriented cutting speed. It puts forward a mathematical model for the optimization of cutting variables in machining.

  10. Evaluation of the hemostatic and coagulation effects of AUTO CUT and DRY CUT using a computer-controlled cutting system.

    Science.gov (United States)

    Szyrach, Mara N I; Tolba, Rene H; Voigtländer, Matthias; Neugebauer, Alexander; Enderle, Markus D

    2010-05-01

    To evaluate a newly developed computer-controlled cutting system for the generation of standardized resections and to systematically compare the hemostatic properties and tissue effect of 2 cutting modes, namely, AUTO CUT and DRY CUT used in urologic procedures. An isolated perfused kidney model was used to assess blood loss and coagulation depth after resection of tissue specimens of standardized geometry, size, and cutting velocity with a resection loop. Three different effect settings (E1, E3, and E6; 200 W) of the electrosurgical modes AUTO CUT and DRY CUT were compared. Blood loss was determined semiquantitatively by weighing a swab before and after placing it onto the resection area. The coagulation depth was estimated microscopically on cross sections. The computer-controlled cutting system creates resections of standardized geometry and size with a high reproducibility. An effect level-dependent increase in hemostasis and coagulation depth could be demonstrated with the cutting modes DRY CUT and AUTO CUT using this computer-controlled cutting system. The hemostatic effect with DRY CUT is significantly more pronounced than with AUTO CUT (E1, E3: P computer-controlled cutting system creating reproducible resections in combination with the isolated perfused kidney model offers the possibility to systematically investigate bleeding rate and coagulation depth. The stronger hemostatic properties of the DRY CUT mode are more favorable for urologic interventions requiring a higher hemostatic effect than the AUTO CUT mode. Copyright 2010 Elsevier Inc. All rights reserved.

  11. Cloning of Human Tumor Necrosis Factor (TNF) Receptor cDNA and Expression of Recombinant Soluble TNF-Binding Protein

    Science.gov (United States)

    Gray, Patrick W.; Barrett, Kathy; Chantry, David; Turner, Martin; Feldmann, Marc

    1990-10-01

    The cDNA for one of the receptors for human tumor necrosis factor (TNF) has been isolated. This cDNA encodes a protein of 455 amino acids that is divided into an extracellular domain of 171 residues and a cytoplasmic domain of 221 residues. The extracellular domain has been engineered for expression in mammalian cells, and this recombinant derivative binds TNFα with high affinity and inhibits its cytotoxic activity in vitro. The TNF receptor exhibits similarity with a family of cell surface proteins that includes the nerve growth factor receptor, the human B-cell surface antigen CD40, and the rat T-cell surface antigen OX40. The TNF receptor contains four cysteine-rich subdomains in the extra-cellular portion. Mammalian cells transfected with the entire TNF receptor cDNA bind radiolabeled TNFα with an affinity of 2.5 x 10-9 M. This binding can be competitively inhibited with unlabeled TNFα or lymphotoxin (TNFβ).

  12. Molecular cloning of the cDNA and chromosomal localization of the gene for a putative seven-transmembrane segment (7-TMS) receptor isolated from human spleen

    Energy Technology Data Exchange (ETDEWEB)

    Federsppiel, B.; Melhado, I.G.; Delaney, A.; Clark-Lewis, I. (Univ. of British Columbia, Vancouver (Canada)); Duncan, A.M.V. (Queens Univ., Kinston, Ontario (Canada)); Jirik, F.R. (Hospital for Sick Children, Toronto, Ontario (Canada))

    1993-06-01

    A family of proinflammatory cytokines sharing several structural features has been described and includes, for example, interleukin-8, monocyte chemoattractant protein-1, and melanocyte growth stimulatory activity. Recently, the receptors for interleukin-8 have been isolated and found to belong to the seven-transmembrane domain class of G protein-coupled receptors. As other members of this cytokine family likely interact with similar receptors, the polymerase chain reaction was employed to isolate related receptors from human peripheral blood adherent cells. Degenerate oligonucleotide primers based on the rabbit interleukin-8 receptor sequence were used. The corresponding full-length cDNA was isolated from a human spleen cDNA library. The predicted protein sequence of this clone, designated pBE1.3, was 93% identical to that of a cDNA isolated from bovine locus coeruleus, which apparently encodes a neuropeptide Y receptor, and also shows similarity with the interleukin-8 receptor and the human cytomegalovirus US28 sequences. The gene, designated D2S201E, was localized to human chromosome 2q21. By Northern blotting, transcripts hybridizing to this cDNA were present in a variety of tissues and cells, including those of hemopoietic origin. 32 refs., 5 figs.

  13. Construction of a rice immature seeds cDNA library and molecular cloning of oryzacystatin cDNA

    Institute of Scientific and Technical Information of China (English)

    周兆斓; 朱祯; 刘春明; 张海涛; 肖桂芳; 李向辉

    1996-01-01

    Total RNA was extracted from rice immature seeds harvested 2 weeks after flowering; then mRNA was purified. cDNA with NotI and SaiI cohesive ends was synthesized and inserted into λgt22A. After packaged in vitno, the cDNA library was constructed with 1.5×106pfu. A 21-mer oligodeoxynucleotide was synthesized according to the 5’-end conserved coding sequence of oryzacystatin (a thiol proteinase inhibitor) and labeled as a probe. From 2.1 × 104 pfu, 9 positive dones have been isolated, 8 of which contain the entire coding region of oryzacystatin. λOC1 has the longest cDNA insert, which contains an open reading frame of 309 bp coding sequence, 84 bp 5’-end non-coding region and a poly(A) signal AATAAA at the 3’-end followed by 31 Nt of poly(A). The coding sequence is the same compared with oryzacystatin genomic DNA sequence, while there are some obvious differences such as insertion and variation in the non-coding region, especially lots of nonsucoessive insertion in the 3’ region after poly(A) signal.

  14. Analysis of expressed sequence tags generated from full-length enriched cDNA libraries of melon

    Directory of Open Access Journals (Sweden)

    Bendahmane Abdelhafid

    2011-05-01

    Full Text Available Abstract Background Melon (Cucumis melo, an economically important vegetable crop, belongs to the Cucurbitaceae family which includes several other important crops such as watermelon, cucumber, and pumpkin. It has served as a model system for sex determination and vascular biology studies. However, genomic resources currently available for melon are limited. Result We constructed eleven full-length enriched and four standard cDNA libraries from fruits, flowers, leaves, roots, cotyledons, and calluses of four different melon genotypes, and generated 71,577 and 22,179 ESTs from full-length enriched and standard cDNA libraries, respectively. These ESTs, together with ~35,000 ESTs available in public domains, were assembled into 24,444 unigenes, which were extensively annotated by comparing their sequences to different protein and functional domain databases, assigning them Gene Ontology (GO terms, and mapping them onto metabolic pathways. Comparative analysis of melon unigenes and other plant genomes revealed that 75% to 85% of melon unigenes had homologs in other dicot plants, while approximately 70% had homologs in monocot plants. The analysis also identified 6,972 gene families that were conserved across dicot and monocot plants, and 181, 1,192, and 220 gene families specific to fleshy fruit-bearing plants, the Cucurbitaceae family, and melon, respectively. Digital expression analysis identified a total of 175 tissue-specific genes, which provides a valuable gene sequence resource for future genomics and functional studies. Furthermore, we identified 4,068 simple sequence repeats (SSRs and 3,073 single nucleotide polymorphisms (SNPs in the melon EST collection. Finally, we obtained a total of 1,382 melon full-length transcripts through the analysis of full-length enriched cDNA clones that were sequenced from both ends. Analysis of these full-length transcripts indicated that sizes of melon 5' and 3' UTRs were similar to those of tomato, but

  15. Cut Front Geometry Characterization in Cutting Applications of Brass with Abrasive Water Jet

    Science.gov (United States)

    Akkurt, Adnan

    2010-06-01

    Abrasive water jet (AWJ) cutting is an advanced manufacturing process for machining hard to cut materials. In this study, brass-353 samples of different thicknesses were cut by AWJ using different feed rates to identify the relationships between depth of cut (material thickness), feed rate, and deflection of cutting edge geometry. The effects of material thickness on the AWJ cut surface roughness were investigated and discussed. Deflection of cutting edge geometry in AWJ cutting process was assessed. Cutting edge geometry was characterized by analyzing the surface properties of cut samples.

  16. [cDNA library construction from panicle meristem of finger millet].

    Science.gov (United States)

    Radchuk, V; Pirko, Ia V; Isaenkov, S V; Emets, A I; Blium, Ia B

    2014-01-01

    The protocol for production of full-size cDNA using SuperScript Full-Length cDNA Library Construction Kit II (Invitrogen) was tested and high quality cDNA library from meristematic tissue of finger millet panicle (Eleusine coracana (L.) Gaertn) was created. The titer of obtained cDNA library comprised 3.01 x 10(5) CFU/ml in avarage. In average the length of cDNA insertion consisted about 1070 base pairs, the effectivity of cDNA fragment insertions--99.5%. The selective sequencing of cDNA clones from created library was performed. The sequences of cDNA clones were identified with usage of BLAST-search. The results of cDNA library analysis and selective sequencing represents prove good functionality and full length character of inserted cDNA clones. Obtained cDNA library from meristematic tissue of finger millet panicle represents good and valuable source for isolation and identification of key genes regulating metabolism and meristematic development and for mining of new molecular markers to conduct out high quality genetic investigations and molecular breeding as well.

  17. Preparation of fluorescent-dye-labeled cDNA from RNA for microarray hybridization.

    Science.gov (United States)

    Ares, Manuel

    2014-01-01

    This protocol describes how to prepare fluorescently labeled cDNA for hybridization to microarrays. It consists of two steps: first, a mixture of anchored oligo(dT) and random hexamers is used to prime amine-modified cDNA synthesis by reverse transcriptase using a modified deoxynucleotide with a reactive amine group (aminoallyl-dUTP) and an RNA sample as a template. Second, the cDNA is purified and exchanged into bicarbonate buffer so that the amine groups in the cDNA react with the dye N-hydroxysuccinimide (NHS) esters, covalently joining the dye to the cDNA. The dye-coupled cDNA is purified again, and the amount of dye incorporated per microgram of cDNA is determined.

  18. Extensions of cutting problems: setups

    Directory of Open Access Journals (Sweden)

    Sebastian Henn

    2013-08-01

    Full Text Available Even though the body of literature in the area of cutting and packing is growing rapidly, research seems to focus on standard problems in the first place, while practical aspects are less frequently dealt with. This is particularly true for setup processes which arise in industrial cutting processes whenever a new cutting pattern is started (i.e. a pattern is different from its predecessor and the cutting equipment has to be prepared in order to meet the technological requirements of the new pattern. Setups involve the consumption of resources and the loss of production time capacity. Therefore, consequences of this kind must explicitly be taken into account for the planning and control of industrial cutting processes. This results in extensions to traditional models which will be reviewed here. We show how setups can be represented in such models, and we report on the algorithms which have been suggested for the determination of solutions of the respective models. We discuss the value of these approaches and finally point out potential directions of future research.

  19. Cutting inlays with a laser

    Science.gov (United States)

    Swaczyna, Irena; Grabczewski, Zbigniew

    1995-03-01

    To cut inlay from a stack of glued veneer a CO2 HEBAR-1A laser was used. For setting optimal working parameters of the set used in industrial production of inlay the following elements were defined: the shape and dimensions of the cutting fissure, the dependence between the width of the cutting fissure and the speed with which the laser beam moves and the total thickness of the stack of veneer sheets, the application of the laser for cutting various patterns. Computer aided designing and computer steering of the laser beam enables fast and precise production of large numbers of inlay elements not only from wood but also from other materials like glass, stone, metal, etc. Taking into consideration the high running cost of such a laser set and its very big production only few factories or even one factory in the given area could produce inlay ready for gluing. Further investigation should be carried out on this field particularly considering the lowering of costs not only in inlay production but generally where cutting of wood is concerned.

  20. Equilibrium Analysis in Cake Cutting

    DEFF Research Database (Denmark)

    Branzei, Simina; Miltersen, Peter Bro

    2013-01-01

    Cake cutting is a fundamental model in fair division; it represents the problem of fairly allocating a heterogeneous divisible good among agents with different preferences. The central criteria of fairness are proportionality and envy-freeness, and many of the existing protocols are designed...... to guarantee proportional or envy-free allocations, when the participating agents follow the protocol. However, typically, all agents following the protocol is not guaranteed to result in a Nash equilibrium. In this paper, we initiate the study of equilibria of classical cake cutting protocols. We consider one...... of the simplest and most elegant continuous algorithms -- the Dubins-Spanier procedure, which guarantees a proportional allocation of the cake -- and study its equilibria when the agents use simple threshold strategies. We show that given a cake cutting instance with strictly positive value density functions...

  1. Cloning and analysis of the mouse Fanconi anemia group a cDNA and an overlapping penta zinc finger cDNA

    NARCIS (Netherlands)

    Wong, JCY; Alon, N; Norga, K; Kruyt, FAE; Youssoufian, H; Buchwald, M

    2000-01-01

    Despite the cloning of four disease-associated genes for Fanconi anemia (FA), the molecular pathogenesis of FA remains largely unknown. To study FA complementation group A using the mouse as a mode I system, we cloned and characterized the mouse homolog of the human FANCA cDNA, The mouse cDNA

  2. Cloning and prokaryotic expression of HGLP cDNA

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A novel cDNA, HGLP, which encodes a G- protein coupled receptor (GPCR) like protein, has been isolated and cloned. The coding region of the human HGLP predicts a 7-transmembrane region protein with motifs of rhodopsin-like GPCR superfamily. Northern blot analysis reveals a 3-kb transcript in various human tissues examined. The N- and C-terminal coding regions of HGLP, which are deduced as non-transmembrane regions, have been amplified by PCR and cloned into pET30a+ vector. Then the recombinant proteins are highly expressed in E. coli.

  3. Application of water jetfor cutting polymer materials

    Directory of Open Access Journals (Sweden)

    A. Stoić

    2013-04-01

    Full Text Available Due to the nature of polymeric materials, during thermal cutting processes it leads to their melting, and therefore appear errors in the final product. This paper presents a “cold” process of cutting polyamide 6 ie. SIPAS 60, where there are given the characteristics of materials and guidelines for satisfactory quality of process. The authors made the cut experiment 32 were they changed the cutting parameters (cutting pressure, cutting feed and abrasive mass flow; the surface roughness was measured by the depth of material, because the roughness changes with the thickness of the material to be cut.

  4. Cutting temperature measurement and material machinability

    Directory of Open Access Journals (Sweden)

    Nedić Bogdan P.

    2014-01-01

    Full Text Available Cutting temperature is very important parameter of cutting process. Around 90% of heat generated during cutting process is then away by sawdust, and the rest is transferred to the tool and workpiece. In this research cutting temperature was measured with artificial thermocouples and question of investigation of metal machinability from aspect of cutting temperature was analyzed. For investigation of material machinability during turning artificial thermocouple was placed just below the cutting top of insert, and for drilling thermocouples were placed through screw holes on the face surface. In this way was obtained simple, reliable, economic and accurate method for investigation of cutting machinability.

  5. Cutting Characteristics of Force Controllable Milling Head

    Institute of Scientific and Technical Information of China (English)

    Shirakashi; Takahiro; Shibuya; Wataru

    2002-01-01

    In order to control cutting force and its direction i n milling operation, a new milling head was developed. The head has two milling cutters, which are connected by a pair of gears and rotate in opposite direction respectively. Both up-cut and down-cut can be carried out simultaneously by t hese milling cutters. The each depth of cut, the ratio of up/down cutting depth , by these cutters can be also selected. The cutting force characteristics were experimentally discussed by changing the ratio. The cut...

  6. The Cutting Process, Chips and Cutting Forces in Machining CFRP

    DEFF Research Database (Denmark)

    Koplev, A.; Lystrup, Aage; Vorm, T.

    1983-01-01

    The cutting of unidirectional CFRP, perpendicular as well as parallel to the fibre orientation, is examined. Shaping experiments, ‘quick-stop’ experiments, and a new chip preparation technique are used for the investigation. The formation of the chips, and the quality of the machined surface...

  7. Molecular cloning of a cysteine proteinase cDNA from adult Ancylostoma ceylanicum by the method of rapid amplification of cDNA ends using polymerase chain reaction.

    Science.gov (United States)

    Mieszczanek, J; Kofta, W; Wedrychowicz, H

    2000-12-01

    The hookworm Ancylostoma ceylanicum is a parasite of great importance in human and veterinary medicine. The most promising vaccination trials against hookworm infections are based on antigens belonging to the proteinase family. The aim of the present research was to isolate a cysteine proteinase gene from A. ceylanicum. This was achieved by rapid amplification of cDNA ends using polymerase chain reaction (RACE-PCR). A set of consensus oligonucleotide primers was designed to anneal to the conserved coding regions of cysteine proteinase. The PCR products were cloned and sequenced. The novel sequence displayed a high degree of homology with genes of cysteine proteinases known from other hookworm species. In the coding region the nucleotide identity with accp-1, the cysteine proteinase gene of A. caninum, reaches 84.3%. Analysis of the expression of acey-1. the cysteine proteinase gene of A. ceylanicum, suggests that it is produced exclusively in the gland cells of either adult worms or blood-feeding stages of A. ceylanicum.

  8. Correlation methods in cutting arcs

    Energy Technology Data Exchange (ETDEWEB)

    Prevosto, L; Kelly, H, E-mail: prevosto@waycom.com.ar [Grupo de Descargas Electricas, Departamento Ing. Electromecanica, Universidad Tecnologica Nacional, Regional Venado Tuerto, Laprida 651, Venado Tuerto (2600), Santa Fe (Argentina)

    2011-05-01

    The present work applies similarity theory to the plasma emanating from transferred arc, gas-vortex stabilized plasma cutting torches, to analyze the existing correlation between the arc temperature and the physical parameters of such torches. It has been found that the enthalpy number significantly influence the temperature of the electric arc. The obtained correlation shows an average deviation of 3% from the temperature data points. Such correlation can be used, for instance, to predict changes in the peak value of the arc temperature at the nozzle exit of a geometrically similar cutting torch due to changes in its operation parameters.

  9. Microbiopsy/precision cutting devices

    Science.gov (United States)

    Krulevitch, Peter A.; Lee, Abraham P.; Northrup, M. Allen; Benett, William J.

    1999-01-01

    Devices for performing tissue biopsy on a small scale (microbiopsy). By reducing the size of the biopsy tool and removing only a small amount of tissue or other material in a minimally invasive manner, the risks, costs, injury and patient discomfort associated with traditional biopsy procedures can be reduced. By using micromachining and precision machining capabilities, it is possible to fabricate small biopsy/cutting devices from silicon. These devices can be used in one of four ways 1) intravascularly, 2) extravascularly, 3) by vessel puncture, and 4) externally. Additionally, the devices may be used in precision surgical cutting.

  10. Revolutions in rapid amplification of cDNA ends: new strategies for polymerase chain reaction cloning of full-length cDNA ends.

    Science.gov (United States)

    Schaefer, B C

    1995-05-20

    Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction (PCR)-based technique which was developed to facilitate the cloning of full-length cDNA 5'- and 3'-ends after a partial cDNA sequence has been obtained by other methods. While RACE can yield complete sequences of cDNA ends in only a few days, the RACE procedure frequently results in the exclusive amplification of truncated cDNA ends, undermining efforts to generate full-length clones. Many investigators have suggested modifications to the RACE protocol to improve the effectiveness of the technique. Based on first-hand experience with RACE, a critical review of numerous published variations of the key steps in the RACE method is presented. Also included is a detailed, effective protocol based on RNA ligase-mediated RACE/reverse ligation-mediated PCR, as well as a demonstration of its utility.

  11. Polymerase reaction without primers throughout for the reconstruction of full-length cDNA from products of rapid amplification of cDNA ends (RACE).

    Science.gov (United States)

    Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Watanabe, Kousuke; Emoto, Noriko; Nagase, Takahide; Ohishi, Nobuya; Takai, Daiya

    2011-07-01

    Rapid amplification of cDNA ends (RACE) has widely been used to determine both ends of the cDNA from its partial sequence. Conventionally, 5'- and 3'-RACE products were ligated at a restriction site in the overlap region to reconstruct the full-length cDNA; however, reconstruction is difficult if no appropriate restriction enzymes are available. Here, we report a novel method to reconstruct full-length cDNA with DNA polymerase. Instead of usual PCR, chain reactions were avoided and the elongation time was shortened, which enables non-specific products or undesired point mutations to be minimized. We successfully reconstructed and TA-cloned a full-length cDNA of echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene variant 2 from RACE products obtained from a surgically resected lung adenocarcinoma sample. We also evaluated some parameters to provide recommendations for this new method.

  12. Advancing family psychology.

    Science.gov (United States)

    Fiese, Barbara H

    2016-02-01

    To realize the broad and complex nature of the field of family psychology, I have slightly revised the mission statement of the Journal of Family Psychology (JFP) to capture contemporary scholarship in family psychology and to advance systems perspectives in this top-tier scientific journal. Over the next 6 years, I hope that authors will consider JFP as an outlet for their best work in the following areas: (1) JFP addresses societal challenges faced by families today; (2) JFP publishes important studies on what makes couple and family relationships work; (3) JFP is a leader in publishing reports that use cutting-edge sophisticated approaches to research design and data analysis; and (4) JFP imparts knowledge about effective therapy and prevention programs relevant to couples and families. The journal is also expanding its publication rate to eight issues per year.

  13. Sequence characterization of a human embryonic craniofacial cDNA library

    Energy Technology Data Exchange (ETDEWEB)

    Padanilam, B.J.; Barsel, S.; Solursh, M. [and others

    1994-09-01

    Broad-based sequencing approaches for the characterization of human cDNA libraries have proven successful in identifying large numbers of novel genes of specific tissue or developmental stages. To pursue our interests in human craniofacial development, stages. To pursue our interests in human craniofacial development, we have made use of both subtracted and unsubtracted cDNA libraries constructed from embryonic craniofacial tissue obtained from pooled samples at 42-54 days gestation. Single-pass sequencing was carried out using an ABI automated sequencer and T3 or T7 primers. Sequences were characterized using BLAST and GRAIL, and the identified homologous sequences grouped according to gene class and family. Four genes have been mapped using repeat sequence elements identified in the clones. Using primers developed from sequence data, other genes are being mapped using a panel of somatic cell hybrids. To date, a total of 786 sequences have been returned with 35% identifying no homologies, and 35% with strong homologies to previously identified genes. A number of genes previously identified to play a role in human embryonic development have been returned from the sequence comparisons providing evidence that the library is representative of this tissue and stage of development. Previous characterization of the library has also identified a number of novel embryonically expressed human homeobox genes. Genes felt to be of special relevance based on their homology to characterized genes known to play a role in development or that are members of novel classes but with high scores on GRAIL searches are being characterized using whole mount in situ hybridization with mouse embryos. Characterization of the library with respect to chromosomal mapping, gene types and make-up, and embryonic expression patterns will be presented.

  14. Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L. Taub

    Directory of Open Access Journals (Sweden)

    Dixon Richard A

    2007-11-01

    Full Text Available Abstract Background Guar, Cyamopsis tetragonoloba (L. Taub, is a member of the Leguminosae (Fabaceae family and is economically the most important of the four species in the genus. The endosperm of guar seed is a rich source of mucilage or gum, which forms a viscous gel in cold water, and is used as an emulsifier, thickener and stabilizer in a wide range of foods and industrial applications. Guar gum is a galactomannan, consisting of a linear (1→4-β-linked D-mannan backbone with single-unit, (1→6-linked, α-D-galactopyranosyl side chains. To better understand regulation of guar seed development and galactomannan metabolism we created cDNA libraries and a resulting EST dataset from different developmental stages of guar seeds. Results A database of 16,476 guar seed ESTs was constructed, with 8,163 and 8,313 ESTs derived from cDNA libraries I and II, respectively. Library I was constructed from seeds at an early developmental stage (15–25 days after flowering, DAF, and library II from seeds at 30–40 DAF. Quite different sets of genes were represented in these two libraries. Approximately 27% of the clones were not similar to known sequences, suggesting that these ESTs represent novel genes or may represent non-coding RNA. The high flux of energy into carbohydrate and storage protein synthesis in guar seeds was reflected by a high representation of genes annotated as involved in signal transduction, carbohydrate metabolism, chaperone and proteolytic processes, and translation and ribosome structure. Guar unigenes involved in galactomannan metabolism were identified. Among the seed storage proteins, the most abundant contig represented a conglutin accounting for 3.7% of the total ESTs from both libraries. Conclusion The present EST collection and its annotation provide a resource for understanding guar seed biology and galactomannan metabolism.

  15. Wedge cutting of mild steel by CO 2 laser and cut-quality assessment in relation to normal cutting

    Science.gov (United States)

    Yilbas, B. S.; Karatas, C.; Uslan, I.; Keles, O.; Usta, Y.; Yilbas, Z.; Ahsan, M.

    2008-10-01

    In some applications, laser cutting of wedge surfaces cannot be avoided in sheet metal processing and the quality of the end product defines the applicability of the laser-cutting process in such situations. In the present study, CO 2 laser cutting of the wedge surfaces as well as normal surfaces (normal to laser beam axis) is considered and the end product quality is assessed using the international standards for thermal cutting. The cut surfaces are examined by the optical microscopy and geometric features of the cut edges such as out of flatness and dross height are measured from the micrographs. A neural network is introduced to classify the striation patterns of the cut surfaces. It is found that the dross height and out of flatness are influenced significantly by the laser output power, particularly for wedge-cutting situation. Moreover, the cut quality improves at certain value of the laser power intensity.

  16. A drosophila full-length cDNA resource

    Energy Technology Data Exchange (ETDEWEB)

    Stapleton, Mark; Carlson, Joseph; Brokstein, Peter; Yu, Charles; Champe, Mark; George, Reed; Guarin, Hannibal; Kronmiller, Brent; Pacleb, Joanne; Park, Soo; Rubin, Gerald M.; Celniker, Susan E.

    2003-05-09

    Background: A collection of sequenced full-length cDNAs is an important resource both for functional genomics studies and for the determination of the intron-exon structure of genes. Providing this resource to the Drosophila melanogaster research community has been a long-term goal of the Berkeley Drosophila Genome Project. We have previously described the Drosophila Gene Collection (DGC), a set of putative full-length cDNAs that was produced by generating and analyzing over 250,000 expressed sequence tags (ESTs) derived from a variety of tissues and developmental stages. Results: We have generated high-quality full-insert sequence for 8,921 clones in the DGC. We compared the sequence of these clones to the annotated Release 3 genomic sequence, and identified more than 5,300 cDNAs that contain a complete and accurate protein-coding sequence. This corresponds to at least one splice form for 40 percent of the predicted D. melanogaster genes. We also identified potential new cases of RNA editing. Conclusions: We show that comparison of cDNA sequences to a high-quality annotated genomic sequence is an effective approach to identifying and eliminating defective clones from a cDNA collection and ensure its utility for experimentation. Clones were eliminated either because they carry single nucleotide discrepancies, which most probably result from reverse transcriptase errors, or because they are truncated and contain only part of the protein-coding sequence.

  17. Analysis of changes in paper cutting forces during the cutting cycle in single-knife guillotine

    OpenAIRE

    Rusin, Agnieszka; Petriaszwili, Georgij

    2013-01-01

    Paper presents the results of changes in the three components of cutting forces of paper stacks cutting during the cutting cycle in single-knife guillotine. The changes of the three components of cutting force at different stages of cutting cycle were analyzed.

  18. How Can I Stop Cutting?

    Science.gov (United States)

    ... have razors near the tub) listen to soothing music that will shift your mood try a breathing exercise try some relaxing yoga exercises previous continue Things to Help You Express the Pain and Deep Emotion Some people cut because the emotions that they ...

  19. How Can I Stop Cutting?

    Science.gov (United States)

    ... previous continue Things to Help You Express the Pain and Deep Emotion Some people cut because the emotions that they ... may be hard for them to recognize these emotions for what they are — like anger, ... out your hurt, anger, or pain using a pen and paper draw the pain ...

  20. Why I like power cuts...

    CERN Multimedia

    Computer Security Team

    2012-01-01

    Accidental power cuts - a permanent nuisance when running accelerators or computing services, since it takes a lot of time to recover from them. While I feel very sorry for those who are under pressure to get their service running again and deeply regret the loss of down-time and availability, I must admit that I like power cuts: power cuts make computers reboot! And rebooting computers at CERN means all the pending software patches are automatically applied.   But don’t think I am egotistic enough to endorse power cuts. Not necessarily! I am already happy if you regularly patch your computer(s) yourself, where regularly means at least once a month: · If you run a centrally or locally managed Windows computer, give that small orange blinking “CMF” icon in the taskbar a chance in the evening to apply all the pending patches. Also, let it initiate a reboot at the end! · If you have a personal computer with your own Windows operating system, ...

  1. cDNA cloning and expression analysis of a mannose-binding lectin from Pinellia pedatisecta

    Indian Academy of Sciences (India)

    Juan Lin; Xuanwei Zhou; Shi Gao; Xiaojun Liu; Weisheng Wu; Xiaofen Sun; Kexuan Tang

    2007-03-01

    Pinellia pedatisecta agglutinin (PPA) is a very basic protein that accumulates in the tuber of P. pedatisecta. PPA is a hetero-tetramer protein of 40 kDa, composed of two polypeptide chains A (about 12 kDa) and two polypeptides chains B (about 12 kDa). The full-length cDNA of PPA was cloned from P. pedatisecta using SMART RACE-PCR technology; it was 1146 bp and contained a 771 bp open reading frame (ORF) encoding a lectin precursor of 256 amino acid residues with a 24 amino acid signal peptide. The PPA precursor contained 3 mannose-binding sites (QXDXNXVXY) and two conserved domains of 43% identity, PPA-DOM1 (polypeptides A) and PPA-DOM2 (polypeptides B). PPA shared varying identities, ranging from 40% to 85%, with mannose-binding lectins from other species of plant families such as Araceae, Alliaceae, Iridaceae, Liliaceae, Amaryllidaceae and Bromeliaceae. Southern blot analysis indicated that ppa belonged to a multi-copy gene family. Expression pattern analysis revealed that ppa expressed in most tested tissues, with high expression being found in spadix, spathe and tuber. Cloning of the ppa gene not only provides a basis for further investigation of its structure, expression and regulatory mechanism, but also enables us to test its potential role in controlling pests and fungal diseases by transferring the gene into plants in the future.

  2. Isolation and characterization of a cDNA from Cuphea lanceolata encoding a beta-ketoacyl-ACP reductase.

    Science.gov (United States)

    Klein, B; Pawlowski, K; Höricke-Grandpierre, C; Schell, J; Töpfer, R

    1992-05-01

    A cDNA encoding beta-ketoacyl-ACP reductase (EC 1.1.1.100), an integral part of the fatty acid synthase type II, was cloned from Cuphea lanceolata. This cDNA of 1276 bp codes for a polypeptide of 320 amino acids with 63 N-terminal residues presumably representing a transit peptide and 257 residues corresponding to the mature protein of 27 kDa. The encoded protein shows strong homology with the amino-terminal sequence and two tryptic peptides from avocado mesocarp beta-ketoacyl-ACP reductase, and its total amino acid composition is highly similar to those of the beta-ketoacyl-ACP reductases of avocado and spinach. Amino acid sequence homologies to polyketide synthase, beta-ketoreductases and short-chain alcohol dehydrogenases are discussed. An engineered fusion protein lacking most of the transit peptide, which was produced in Escherichia coli, was isolated and proved to possess beta-ketoacyl-ACP reductase activity. Hybridization studies revealed that in C. lanceolata beta-ketoacyl-ACP reductase is encoded by a small family of at least two genes and that members of this family are expressed in roots, leaves, flowers and seeds.

  3. Quality Analysis of Cutting Steel Using Laser

    Directory of Open Access Journals (Sweden)

    Vladislav Markovič

    2013-02-01

    Full Text Available The article explores the quality dependence of the edge surface of steel C45 LST EN 10083-1 obtained cutting the material using laser on different cutting regimes and variations in the thickness of trial steel. The paper presents the influence of the main modes of laser cutting equipment Trulaser 3030, including cutting speed, pressure, angle and the thickness of the surface on the quality characteristics of the sample. The quality of the edge after laser cutting is the most important indicator influencing such technological spread in industry worldwide. Laser cutting is the most popular method of material cutting. Therefore, the article focuses on cutting equipment, cutting defects and methods of analysis. Research on microstructure, roughness and micro-toughness has been performed with reference to edge samples. At the end of the publication, conclusions are drawn.Article in Lithuanian

  4. Quality Analysis of Cutting Steel Using Laser

    Directory of Open Access Journals (Sweden)

    Vladislav Markovič

    2012-12-01

    Full Text Available The article explores the quality dependence of the edge surface of steel C45 LST EN 10083-1 obtained cutting the material using laser on different cutting regimes and variations in the thickness of trial steel. The paper presents the influence of the main modes of laser cutting equipment Trulaser 3030, including cutting speed, pressure, angle and the thickness of the surface on the quality characteristics of the sample. The quality of the edge after laser cutting is the most important indicator influencing such technological spread in industry worldwide. Laser cutting is the most popular method of material cutting. Therefore, the article focuses on cutting equipment, cutting defects and methods of analysis. Research on microstructure, roughness and micro-toughness has been performed with reference to edge samples. At the end of the publication, conclusions are drawn.Article in Lithuanian

  5. Fibre laser cutting stainless steel: Fluid dynamics and cut front morphology

    Science.gov (United States)

    Pocorni, Jetro; Powell, John; Deichsel, Eckard; Frostevarg, Jan; Kaplan, Alexander F. H.

    2017-01-01

    In this paper the morphology of the laser cut front generated by fibre lasers was investigated by observation of the 'frozen' cut front, additionally high speed imaging (HSI) was employed to study the fluid dynamics on the cut front while cutting. During laser cutting the morphology and flow properties of the melt film on the cut front affect cut quality parameters such as cut edge roughness and dross (residual melt attached to the bottom of the cut edge). HSI observation of melt flow down a laser cutting front using standard cutting parameters is experimentally problematic because the cut front is narrow and surrounded by the kerf walls. To compensate for this, artificial parameters are usually chosen to obtain wide cut fronts which are unrepresentative of the actual industrial process. This paper presents a new experimental cutting geometry which permits HSI of the laser cut front using standard, commercial parameters. These results suggest that the cut front produced when cutting medium section (10 mm thick) stainless steel with a fibre laser and a nitrogen assist gas is covered in humps which themselves are covered by a thin layer of liquid. HSI observation and theoretical analysis reveal that under these conditions the humps move down the cut front at an average speed of approximately 0.4 m/s while the covering liquid flows at an average speed of approximately 1.1 m/s, with an average melt depth at the bottom of the cut zone of approximately 0.17 mm.

  6. Characterization of cDNA for PMT: a Partial Nicotine Biosynthesis-Related Gene Isolated from Indonesian Local Tobacco (Nicotiana tabacum cv. Sindoro1

    Directory of Open Access Journals (Sweden)

    Sesanti Basuki

    2013-12-01

    Full Text Available Nicotine is the major alkaloid compound in cultivated tobacco (Nicotiana tabacum that could potentially be converted into carcinogenic compound (nor-nicotine. The PMT gene encoding putrescine N-methyltransferase (PMT is one of the two key genes that play a prominent role in nicotine biosynthesis. The aimed of this study was to isolate and characterize the cDNA sequence originated from Indonesian local tobacco cv. Sindoro1 (Ntpmt_Sindoro1. The results showed that the Ntpmt_Sindoro1 was 1124 bp in length. This cDNA fragment encodes for 374 amino acid residues. The predicted polypeptide from the cDNA is a hidrophilic protein, and has a predicted molecular weight of 40.95 kD. The predicted amino acids sequence also showed high similarity to the PMT gene product Nicotiana sp. available in the GenBank data base. The amino acid sequences also exert conserved residues specifically exhibited only by PMT gene originated from N. tabacum. Clustering analysis revealed that Ntpmt_Sindoro1 belongs to the same clade as the PMT3 gene, a member of the N. tabacum PMT gene family. The Ntpmt_Sindoro1 cDNA sequence covering exon1-exon8 of the PMT gene fragment has been registered in the GenBank data base, under the accession number JX978277.

  7. Rabbit muscle creatine phosphokinase. CDNA cloning, primary structure and detection of human homologues.

    Science.gov (United States)

    Putney, S; Herlihy, W; Royal, N; Pang, H; Aposhian, H V; Pickering, L; Belagaje, R; Biemann, K; Page, D; Kuby, S

    1984-12-10

    A cDNA library was constructed from rabbit muscle poly(A) RNA. Limited amino acid sequence information was obtained on rabbit muscle creatine phosphokinase and this was the basis for design and synthesis of two oligonucleotide probes complementary to a creatine kinase cDNA sequence which encodes a pentapeptide. Colony hybridizations with the probes and subsequent steps led to isolation of two clones, whose cDNA segments partially overlap and which together encode the entire protein. The primary structure was established from the sequence of two cDNA clones and from independently determined sequences of scattered portions of the polypeptide. The reactive cysteine has been located to position 282 within the 380 amino acid polypeptide. The rabbit cDNA hybridizes to digests of human chromosomal DNA. This reveals a restriction fragment length polymorphism associated with the human homologue(s) which hybridizes to the rabbit cDNA.

  8. Laser Cutting of Thin Nickel Bellows

    Science.gov (United States)

    Butler, C. L.

    1986-01-01

    Laser cutting technique produces narrow, precise, fast, and repeatable cuts in thin nickel-allow bellows material. Laser cutting operation uses intense focused beam to melt material and assisting gas to force melted material through part thickness, creating void. When part rotated or moved longitudinally, melting and material removal continuous and creates narrow, fast, precise, and repeatable cut. Technique used to produce cuts of specified depths less than material thickness. Avoids distortion, dents, and nicks produced in delicate materials during lathe trimming operations, which require high cutting-tool pressure and holding-fixture forces.

  9. Cutting

    Science.gov (United States)

    ... way some people try to cope with the pain of strong emotions, intense pressure , or upsetting relationship problems . They may ... with difficulties, even big problems and terrible emotional pain. The help of ... or overwhelming emotions. For other tough situations or strong emotions, it ...

  10. Molecular cloning of growth hormone encoding cDNA of Indian major carps by a modified rapid amplification of cDNA ends strategy

    Indian Academy of Sciences (India)

    T Venugopal; S Mathavan; T J Pandian

    2002-06-01

    A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in Escherichia coli. These sequences show 96–98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein.

  11. Subtractive cDNA cloning using oligo(dT)30-latex and PCR: isolation of cDNA clones specific to undifferentiated human embryonal carcinoma cells.

    OpenAIRE

    1991-01-01

    The human embryonal carcinoma cell line NEC14 can be induced to differentiate by the addition of 10(-2)M N,N'-hexamethylene-bis-acetamide (HMBA). A subtractive cDNA library specific to undifferentiated NEC14 cells was constructed using oligo(dT)30-Latex and polymerase chain reaction (PCR). The method was designed to improve the efficiency of subtraction and the enrichment of cDNA clones corresponding to low abundance mRNAs. The single strand of cDNA was made from mRNA prepared from the HMBA-t...

  12. Molecular cloning of growth hormone encoding cDNA of Indian major carps by a modified rapid amplification of cDNA ends strategy.

    Science.gov (United States)

    Venugopal, T; Mathavan, S; Pandian, T J

    2002-06-01

    A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in Escherichia coli. These sequences show 96-98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein.

  13. Effect of cutting parameters on chip formation in orthogonal cutting

    Directory of Open Access Journals (Sweden)

    S. Ben Salem

    2012-01-01

    Full Text Available Purpose: of this paper is to study the chip formation to obtain the optimal cutting conditions and to observe the different chip formation mechanisms. Analysis of machining of a hardened alloy, X160CrMoV12-1 (cold work steel: AISI D2 with a ferritic and cementite matrix and coarse primary carbides, showed that there are relationships between the chip geometry, cutting conditions and the different micrographs under different metallurgical states.Design/methodology/approach: Machining of hardened alloys has some metallurgical and mechanical difficulties even if many successful processes have been increasingly developed. A lot of study has been carried out on this subject, however only with modest progress showing specific results concerning the real efficiency of chip formation. Hence, some crucial questions remain unanswered: the evolution of white layers produced during progressive tool flank wear in dry hard turning and to correlate this with the surface integrity of the machined surface. For the experimental study here, various cutting speeds and feed rates have been applied on the work material.Findings: The “saw-tooth type chips” geometry has been examined and a specific attention was given to the chip samples that were metallographically processed and observed under scanning electronic microscope (SEM to determine if white layers are present.Research limitations/implications: This research will be followed by a detail modelling and need more experimental results for a given a good prediction of the results occurred on the damage related to the microstructure by using the cutting parameters.Practical implications: A special detail was given to the mechanism of chip formation resulting from hard machining process and behaviour of steel at different metallurgical states on the material during the case of annealing and or the case of quench operations.Originality/value: For the sake of simplicity, ANOVA (Analysis of Variance was used to

  14. Preparation of microbial community cDNA for metatranscriptomic analysis in marine plankton.

    Science.gov (United States)

    Stewart, Frank J

    2013-01-01

    High-throughput sequencing and analysis of microbial community cDNA (metatranscriptomics) are providing valuable insight into in situ microbial activity and metabolism in the oceans. A critical first step in metatranscriptomic studies is the preparation of high-quality cDNA. At the minimum, preparing cDNA for sequencing involves steps of biomass collection, RNA preservation, total RNA extraction, and cDNA synthesis. Each of these steps may present unique challenges for marine microbial samples, particularly for deep-sea samples whose transcriptional profiles may change between water collection and RNA preservation. Because bacterioplankton community RNA yields may be relatively low (microbiology research.

  15. A method for generating subtractive cDNA libraries retaining clones containing repetitive elements.

    OpenAIRE

    1997-01-01

    Here we describe a two-stepped photobiotin-based procedure to enrich a target (canine retinal) cDNA library for tissue specific clones without removing those containing repetitive ( SINE ) elements, despite the presence of these elements in the driver population. In a first hybridization excess SINE elements were hybridized to a driver (canine cerebellar) cDNA. In a second hybridization target cDNA was added to this reaction. The resulting cDNA library was enriched for retinal specific clones...

  16. Influence of the Magnetic High-speed Steel Cutting Tool on Cutting Capability

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The high-speed steel cutting tool has advantaged i n modern cutting tool for its preferable synthetical performance, especially, in a pplication of complicated cutting tools. Therefore, the study of the high-speed steel cutting tools that occupied half of cutting tools has become an importa nt way of studying on modern cutting technology. The cutting performance of hi gh speed-steel cutting tools will be improved by magnetization treating method. Microstructure of high-speed steel will be changed as a ...

  17. Soluble-oil cutting fluid

    Energy Technology Data Exchange (ETDEWEB)

    Rawlinson, A.P.; Whitby, R.D.; White, J.

    1989-04-11

    A composition for the preparation of a soluble oil for use in a cutting fluid comprises a mineral oil and, as an emuslifier, an effective amount of a sulfonate of a branched polymer of a C/sub 3/ to C/sub 5/ olefin. Preferably the polyolefin chain of the sulphonate has an average molecular weight in the range 275 to 560 and the polyolefin is polyisobutene. A soluble oil can be prepared from the above composition by the addition of a conventional corrosion inhibitor and diluted with water to make a cutting fluid. Advantages of the novel emulsifier are that it is resistant to breakdown by micro-organisms and does not require the addition of a coupling agent.

  18. Lysosomal {beta}-mannosidase: cDNA cloning and characterization

    Energy Technology Data Exchange (ETDEWEB)

    Chen, H.; Leipprandt, J.R.; Traviss, C.E. [Michigan State Univ., East Lansing, MI (United States)] [and others

    1994-09-01

    Lysosomal {beta}-mannosidase is an exoglycosidase that cleaves the single {beta}-linked mannose residue from the non-reducing end of all N-linked glycoprotein oligosaccharides. Deficiency of this enzyme results in {beta}-mannosidosis, a severe neurodegenerative disease in goats and cattle. The human cases described have a milder, highly variable presentation. Study of the molecular pathology of this disease in ruminants and humans and development of the animal model for gene therapy studies required cloning of the gene for {beta}-mannosidase has been cloned. {beta}-Mannosidase cDNA were obtained from a bovine thyroid cDNA library by screening with mixed oligonucleotides derived from peptide sequences resulting from microsequencing of bovine {beta}-mannosidase peptides. A total of six independent positive clones were identified from 5 x 10{sup 5} plaques, covering about 80% of the C-terminal region. The missing 5{prime} region was obtained using 5{prime} RACE. The full-length construct contains 3852-bp nucleotides, encoding 879 amino acids. The initiation codon is followed by 17 amino acids containing the characteristics of a typical signal peptide sequence. The deduced amino acid sequence is colinear with all peptide sequences determined by protein microsequencing. Northern blot analysis demonstrated a 4.2 kb single transcript in various tissues from both normal and affected goats and calves. The mRNA level was decreased in affected {beta}-mannosidosis animals. The gene encoding {beta}-mannosidase was localized on human chromosome 4 by Southern analysis of rodent/human somatic cell hybrids. The mutation in bovine {beta}-mannosidosis has been identified. This is the first report of cloning of the {beta}-mannosidase gene.

  19. Amibiasis cutánea

    OpenAIRE

    Saravia Gómez, José; Rocha Posada, Hernando; Cortés B., Carlos; Ríos, Aníbal

    2011-01-01

    Los autores hacen un análisis de la etiopatogenia, de la anatomía patológica y del cuadro clínico de la amibiasis cutánea. Valoran los diferentes métodos de diagnóstico, el diagnóstico diferencial y el tratamiento de la entidad. Ilustran el trabajo con la presentación de un caso.

  20. Drill cuttings mount formation study

    Science.gov (United States)

    Teh, Su Yean; Koh, Hock Lye

    2014-07-01

    Oil, Gas and Energy sector has been identified as an essential driving force in the Malaysian Economic Transformation Programs (ETP). Recently confirmed discovery of many offshore oil and gas deposits in Malaysian waters has ignited new confidence in this sector. However, this has also spurred intense interest on safeguarding the health and environment of coastal waters in Malaysia from adverse impact resulting from offshore oil and gas production operation. Offshore discharge of spent drilling mud and rock cuttings is the least expensive and simplest option to dispose of large volumes of drilling wastes. But this onsite offshore disposal may have adverse environmental impacts on the water column and the seabed. It may also pose occupational health hazards to the workers living in the offshore platforms. It is therefore important to model the transport and deposition of drilling mud and rock cuttings in the sea to enable proper assessment of their adverse impacts on the environment and the workers. Further, accumulation of drill particles on the seabed may impede proper operation of pipelines on the seabed. In this paper, we present an in-house application model TUNA-PT developed to cater to local oil and gas industry needs to simulate the dispersion and mount formation of drill cuttings by offshore oil and gas exploration and production platforms. Using available data on Malaysian coastal waters, simulation analyses project a pile formation on the seabed with a maximum height of about 1 m and pile radius of around 30 to 50 m. Simulated pile heights are not sensitive to the heights of release of the cuttings as the sensitivity has been mitigated by the depth of water.

  1. Reservoir management cost-cutting

    Energy Technology Data Exchange (ETDEWEB)

    Gulati, M.S.

    1996-12-31

    This article by Mohinder S. Gulati, Chief Engineer, Unocal Geothermal Operations, discusses cost cutting in geothermal reservoir management. The reservoir engineer or geoscientist can make a big difference in the economical outcome of a project by improving well performance and thus making geothermal energy more competitive in the energy marketplace. Bringing plants online in less time and proving resources to reduce the cycle time are some of the ways to reduce reservoir management costs discussed in this article.

  2. Evaluation of water-jet-assisted cutting

    National Research Council Canada - National Science Library

    1987-01-01

    Recent studies show that the addition of a water jet to the cutting bit can improve cutting performance in harder materials, significantly reduce respirable dust and bit wear, and increase particle size...

  3. European labs fight back against cuts

    CERN Multimedia

    König, R

    1997-01-01

    Germany's 1997 budget contains cuts amounting to 3.7% in funding of domestic research programs and in contributions to international labs. Contributions will be cut to the European Space Agency, the European Synchrotron facility and CERN.

  4. Eliminate Sweetened Drinks, Cut Kids' Sugar Intake

    Science.gov (United States)

    ... fullstory_161163.html Eliminate Sweetened Drinks, Cut Kids' Sugar Intake Average U.S. child consumes about 80 grams ... Looking for the quickest way to cut added sugar from your kid's diet? Eliminate sugar-sweetened drinks, ...

  5. Chip-ejection interference in cutting processes of modern cutting tools

    Institute of Scientific and Technical Information of China (English)

    师汉民

    1999-01-01

    Based on the “principle of minimum energy”, the basic characteristics of non-free cutting are studied; the phenomenon and the nature of chip-ejection interference commonly existing in the cutting process of modem cutting tools are explored. A "synthesis method of elementary cutting tools" is suggested for modeling the cutting process of modem complex cutting tools. The general equation governing the chip-ejection motion is deduced. Real examples of non-free cutting are analyzed and the theoretically predicted results are supported by the experimental data or facts. The sufficient and necessary conditions for eliminating chip-ejection interference and for realizing free cutting are given; the idea and the technical approach of "the principle of free cutting" are also discussed, and a feasible way for improving or optimizing the cutting performance of modem cutting tools is, therefore, found.

  6. THE CLONING OF HRNT-1 USING A COMBINATION OF cDNA LIBRARY SCREENING WITH BIOTIN-LABELED PROBE AND RAPID AMPLIFICATION OF cDNA ENDS

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective: To clone the human counterpart of rat ZA73, EST cloned from rat tracheal epithelial (RTE) neoplastic transformed cell model induced by (a-particles radiation by using mRNA differential display. Methods: According to the sequence of rat ZA73, a probe was biotin-labeled to screen human cDNA library, and then the gene sequence was extended by RACE (rapid amplification of cDNA ends). Result: Human gene HRNT-1 (GenBank Accession Number: AF223393) is 4.256 kb in length, with an ORF located in the region between 254 and 3013 bp. 5' UTS (untranslated sequences) is 253 bp, 3' UTS is 1243 bp. Conclusion: The combination of cDNA library screening with biotin-labeled probes and RACE is an effective method to clone full-length cDNA, especially for sequences longer than 2 kb.

  7. Study on Ceramic Cutting by Plasma Arc

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Engineering ceramics are typical difficult-to-machine materials because of high hardness and brittleness. PAC (Plasma Arc Cutting) is a very important thermal cutting process and has been successfully used in cutting stainless steel and other difficult-to-machine alloys. PAC's application in cutting ceramics, however, is still limited because the most ceramics are not good electronic conducts, and transferred plasma arc cannot be produced between cathode and work-piece. So we presented a method of plasma ...

  8. Application of an improved cDNA competition technique to identify prostate cancer-associated gene.

    Science.gov (United States)

    Rinaldy, A R; Steiner, M S

    1999-11-01

    A technique to improve cDNA library screening was developed by using mixed probes derived from two closely related cDNA populations of high-metastatic MAT-LyLu and low-metastatic AT-1 Dunning R3227 rat prostate cancer sublines. The technique required the generation of a cDNA library from each subline followed by polymerase chain reaction (PCR) amplification of the cDNA insert population. The PCR products derived from the first library were radiolabeled and mixed with an excess amount of PCR products from the second library. The mixture and an excess amount of both the lambda and pBluescript DNA were used as a probe to screen the first cDNA library. This mixed probe (designated the competition probe) differentially cross-hybridized with the plaque lift of the screened first cDNA library. Weak radioactive signals indicated the cross-hybridization of cDNA sequences common to the competition probe mixture and the first cDNA library, whereas strong signals implied unhybridized unique or abundant cDNA sequences in the first cDNA library. The reproducibility of this technique was confirmed by showing that the full-length cDNA clones were associated with the phenotype of the screened first cell line. The isolated clones were characterized as rat nucleolar protein, rat mitochondrial genes coding for 16S and 12S rRNAs, and rat tRNAs specific for valine and phenyl-alanine. This result is consistent with the fact that the first cell line, MAT-LyLu, is metabolically more active than are AT-1 cells because of higher gene dosage or amplification of nucleolar and mitochondrial RNA and its associated genes. Another clone which had a strong signal represented a novel gene associated with the MAT-LyLu cancer phenotype.

  9. Air in xylem vessels of cut flowers

    NARCIS (Netherlands)

    Nijsse, J.; Meeteren, van U.; Keijzer, C.J.

    2000-01-01

    Until now all studies on the role of air emboli in the water uptake of cut flowers describe indirect methods to demonstrate the presencFe of air in the plant tissues. Using cut chrysanthemum flowers, this report is the first one that directly visualises both air and water in xylem ducts of cut

  10. Spending and Cutting are Two Different Worlds

    DEFF Research Database (Denmark)

    Houlberg, Kurt; Olsen, Asmus Leth; Holm Pedersen, Lene

    2016-01-01

    deserving receive the highest cuts. Ideology matters as left-wing councillors prefer more vague categories when cutting and prioritise childcare and unemployment policies when increasing spending. In contrast, right-wing councillors prefer to cut administration and increase spending on roads....

  11. Optical Cutting Interruption Sensor for Fiber Lasers

    Directory of Open Access Journals (Sweden)

    Benedikt Adelmann

    2015-09-01

    Full Text Available We report on an optical sensor system attached to a 4 kW fiber laser cutting machine to detect cutting interruptions. The sensor records the thermal radiation from the process zone with a modified ring mirror and optical filter arrangement, which is placed between the cutting head and the collimator. The process radiation is sensed by a Si and InGaAs diode combination with the detected signals being digitalized with 20 kHz. To demonstrate the function of the sensor, signals arising during fusion cutting of 1 mm stainless steel and mild steel with and without cutting interruptions are evaluated and typical signatures derived. In the recorded signals the piercing process, the laser switch on and switch off point and waiting period are clearly resolved. To identify the cutting interruption, the signals of both Si and InGaAs diodes are high pass filtered and the signal fluctuation ranges being subsequently calculated. Introducing a correction factor, we identify that only in case of a cutting interruption the fluctuation range of the Si diode exceeds the InGaAs diode. This characteristic signature was successfully used to detect 80 cutting interruptions of 83 incomplete cuts (alpha error 3.6% and system recorded no cutting interruption from 110 faultless cuts (beta error of 0. This particularly high detection rate in combination with the easy integration of the sensor, highlight its potential for cutting interruption detection in industrial applications.

  12. 27 CFR 6.92 - Newspaper cuts.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Newspaper cuts. 6.92 Section 6.92 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS âTIED-HOUSEâ Exceptions § 6.92 Newspaper cuts. Newspaper cuts, mats, or...

  13. Improving rooting uniformity in rose cuttings

    NARCIS (Netherlands)

    Telgen, van H.J.; Eveleens-Clark, B.A.; Garcia Victoria, N.

    2007-01-01

    Studies to improve rooting uniformity of single node stem cuttings for rose are reported. We found that the variation in shoot growth in a young rose crop depended on the variation in root number of the cuttings, which, in turn, was related to the auxin concentration applied to the cutting before ro

  14. Global Identification of Significantly Expressed Genes in Developing Endosperm of Rice by Expression Sequence Tags and cDNA Array Approaches

    Institute of Scientific and Technical Information of China (English)

    Qichao Tu; Haitao Dong; Haigen Yao; Yongqi Fang; Cheng'en Dai; Hongmei Luo; Jian Yao; Dong Zhao; Debao Li

    2008-01-01

    Rice endosperm plays a very important role in seedling germination and determines the qualities of fice grain.Although studies on specific gene categories in endosperm have been carried out,global view of gene expression at a transcription level in rice endosperm is still limited.To gain a better understanding of the global and tissue-specific gene expression profiles in rice endosperm,a cDNA library from rice endosperm of immature seeds was sequenced.A cDNA array was constructed based on the tentative unique transcripts derived from expression sequence tag (EST) assembling results and then hybridized with cONAs from five different tissues or organs including endosperm,embryo,leaf,stem and root of rice.Significant redundancy was found for genes encoding prolamin,glutelin,allergen,and starch synthesis proteins,accounting for~34% of the total ESTs obtained.The cDNA array revealed 87 significantly expressed genes in endosperm compared with the other four organs or tissues.These genes included 13 prolamin family proteins,17 glutelin family proteins,12 binding proteins,nine catalytic proteins and four ribosomal proteins,indicating a complicated biological processing in rice endosperm.In addition,Northern verification of 1,4-alpha-glucan branching enzyme detected two isoforms in rice endosperm,the larger one of which only existed in endosperm.

  15. Cloning of a cDNA encoding the smallest neurofilament protein from the rat

    NARCIS (Netherlands)

    J-P. Julien (Jean-Pierre); K. Ramachadran; F.G. Grosveld (Frank)

    1985-01-01

    textabstractWe have cloned a cDNA coding for the smallest rat neurofilament protein. The cDNA is 861 nucleotides long coding for 287 amino acids from the internal alpha-helical region and the carboxy-terminal tail domain of the neurofilament protein. Comparison of the porcine, mouse and rat neurofil

  16. Cloning, sequencing and expression of cDNA encoding growth hormone from Indian catfish (Heteropneustes fossilis)

    Indian Academy of Sciences (India)

    Vikas Anathy; Thayanithy Venugopal; Ramanathan Koteeswaran; Thavamani J Pandian; Sinnakaruppan Mathavan

    2001-09-01

    A tissue-specific cDNA library was constructed using polyA+ RNA from pituitary glands of the Indian catfish Heteropneustes fossilis (Bloch) and a cDNA clone encoding growth hormone (GH) was isolated. Using polymerase chain reaction (PCR) primers representing the conserved regions of fish GH sequences the 3′ region of catfish GH cDNA (540 bp) was cloned by random amplification of cDNA ends and the clone was used as a probe to isolate recombinant phages carrying the full-length cDNA sequence. The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5′ and 3′ untranslated regions of the cDNA are 58 bp and 456 bp long, respectively. The predicted amino acid sequence of H. fossils GH shared 98% homology with other catfishes. Mature GH protein was efficiently expressed in bacterial and zebrafish systems using appropriate expression vectors. The successful expression of the cloned GH cDNA of catfish confirms the functional viability of the clone.

  17. Cloning, sequencing and expression of cDNA encoding growth hormone from Indian catfish (Heteropneustes fossilis)

    Indian Academy of Sciences (India)

    Vikas Anathy; Thayanithy Venugopal; Ramanathan Koteeswaran; Thavamani J Pandian; Sinnakaruppan Mathavan

    2013-03-01

    A tissue-specific cDNA library was constructed using polyA+ RNA from pituitary glands of the Indian catfish Heteropneustes fossilis (Bloch) and a cDNA clone encoding growth hormone (GH) was isolated. Using polymerase chain reaction (PCR) primers representing the conserved regions of fish GH sequences the 3′ region of catfish GH cDNA (540 bp) was cloned by random amplification of cDNA ends and the clone was used as a probe to isolate recombinant phages carrying the full-length cDNA sequence. The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5′ and 3′ untranslated regions of the cDNA are 58 bp and 456 bp long, respectively. The predicted amino acid sequence of H. fossils GH shared 98% homology with other catfishes. Mature GH protein was efficiently expressed in bacterial and zebrafish systems using appropriate expression vectors. The successful expression of the cloned GH cDNA of catfish confirms the functional viability of the clone.

  18. IDENTIFICATION OF DIFFERENTIAL GENES IN OVARIAN CANCER USING REPRESENTATIONAL DIFFERENCE ANALYSIS OF cDNA

    Institute of Scientific and Technical Information of China (English)

    Hong Chen; Min Wang; Xin-yan Wang; Shan Gao; Jun Wang; Xiao-ming Guan

    2005-01-01

    Objective To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA (cDNA-RDA). Methods cDNA-RDA was performed to identify the differentially expressed sequences between cDNAs from cancer tissue and cDNAs from normal ovarian tissue in the same patient who was in the early stage of ovarian serous cystadeno carcinoma. These differentially expressed fragments were cloned and analyzed, then sequenced and compared with known genes.Results Three differentially expressed cDNA fragments were isolated using cDNA from normal ovarian tissue as tester and cDNA from cancer tissue as driver amplicon by cDNA-RDA. DP Ⅲ-1 and DP Ⅲ-2 cDNA clone showed significant ho mology to the cDNA of alpha actin gene; DPⅢ-3 cDNA clone showed significant homology to the cDNA of transgelin gene. Conclusion cDNA-RDA can be used to sensitively identify the differentially expressed genes in ovarian serous cystadenocarcinoma. Ovarian serous cystadenocarcinoma involves alteration of multiple genes.

  19. Construction and characterization of a normalized whole-life-cycle cDNA library of rice

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A cDNA library with genomic complete coverage is a powerful tool for functional genomic studies. For studying the functions of rice genes on a large scale, a normalized whole-life-cycle cDNA library is constructed based on the strategy of saturation hybridization with genomic DNA using rice cultivar Minghui 63, an elite restorer line for a number of rice hybrids that are widely cultivated in China. This library consists of cDNA from 15 directionally cloned cDNA libraries constructed with different tissues from 9 developmental stages. For normalization, the denatured plasmids purified from the 15 directionally cloned libraries are mixed and hybridized with saturated genomic DNA labeled with magnetic beads in two complementary systems. Well-matched plasmids are captured from the hybridized genomic DNA and electroporated into competent DH10B E. coli for construction of the normalized whole-life-cycle cDNA library. This library consists of 62000 clones with an average insert length about 1.4 kb. Inverse Northern blotting shows that this cDNA library included many rarely expressed genes and tissue-specific genes. Sequencing of 10750 cDNA clones of this library reveals 6399 unique ESTs (expressed sequence tags), indicating that the non-redundancy of the library is about 59.5%. This library has been used to make cDNA microarrays for functional genomic studies.

  20. Peptidomics combined with cDNA library unravel the diversity of centipede venom

    DEFF Research Database (Denmark)

    Rong, Mingqiang; Yang, Shilong; Wen, Bo

    2015-01-01

    of centipede venom. In the present study, we use peptidomics combined with cDNA library to uncover the diversity of centipede Scolopendra subspinipes mutilans L. Koch. 192 peptides were identified by LC-MS/MS and 79 precursors were deduced by cDNA library. Surprisingly, the signal peptides of centipede toxins...

  1. Genomic and cDNA cloning of a novel mouse lipoxygenase gene

    NARCIS (Netherlands)

    Willems van Dijk, K.; Steketee, K.; Havekes, L.; Frants, R.; Hofker, M.

    1995-01-01

    A novel 12- and 15-lipoxygenase related gene was isolated from a mouse strain 129 genomic phage library in a screen with a human 15-lipoxygenase cDNA probe. The complete genomic sequence revealed 14 exons and 13 introns covering 7.3 kb of DNA. The splice junctions were verified from the cDNA

  2. cDNA library information - Dicty_cDB | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available (C) 5) sexually fusion-competent KAX3 cells (Gamete phase) (F) cDNA library construction method How to const...dir) 2) Full-length cDNA libraries (oligocapped method)(fl) 3) Gamete-specific subtraction library (sub) cDN

  3. Automated Laser Cutting In Three Dimensions

    Science.gov (United States)

    Bird, Lisa T.; Yvanovich, Mark A.; Angell, Terry R.; Bishop, Patricia J.; Dai, Weimin; Dobbs, Robert D.; He, Mingli; Minardi, Antonio; Shelton, Bret A.

    1995-01-01

    Computer-controlled machine-tool system uses laser beam assisted by directed flow of air to cut refractory materials into complex three-dimensional shapes. Velocity, position, and angle of cut varied. In original application, materials in question were thermally insulating thick blankets and tiles used on space shuttle. System shapes tile to concave or convex contours and cuts beveled edges on blanket, without cutting through outer layer of quartz fabric part of blanket. For safety, system entirely enclosed to prevent escape of laser energy. No dust generated during cutting operation - all material vaporized; larger solid chips dislodged from workpiece easily removed later.

  4. Isolation and characterization of a full-length cDNA coding for an adipose differentiation-related protein.

    OpenAIRE

    Jiang, H P; Serrero, G

    1992-01-01

    We have previously isolated from a 1246 adipocyte cDNA library a cDNA clone called 154, corresponding to a mRNA that increases abundantly at a very early time during the differentiation of 1246 adipocytes and in adipocyte precursors in primary culture. We show here that the mRNA encoded by this cDNA is expressed abundantly and preferentially in mouse fat pads. A full-length cDNA for clone 154 was isolated by the RACE (rapid amplification of cDNA ends) protocol. Sequence analysis of this cDNA ...

  5. Isolation and characterization of a full-length cDNA coding for an adipose differentiation-related protein

    OpenAIRE

    Jiang, Hui-Ping; Serrero, Ginette

    1992-01-01

    We have previously isolated from a 1246 adipocyte cDNA library a cDNA clone called 154, corresponding to a mRNA that increases abundantly at a very early time during the differentiation of 1246 adipocytes and in adipocyte precursors in primary culture. We show here that the mRNA encoded by this cDNA is expressed abundantly and preferentially in mouse fat pads. A full-length cDNA for clone 154 was isolated by the RACE (rapid amplification of cDNA ends) protocol. Sequence analysis of this cDNA ...

  6. Effect of Type of Cutting Tips on Cutting Forces in Turning

    Directory of Open Access Journals (Sweden)

    Polák Pavel

    2014-10-01

    Full Text Available The aim of this article is to demonstrate the efect of cutting materials and geometry of cutting tips on cutting forces in turning as well as the quality and precision of machined surface. The experiment focuses on measuring cutting forces when turning a sample of steel 11 523 at a constant feed rate and cut depth and at varying speeds of a spindle. Measurements were made using exchangeable cutting tips of diferent types. The results will be evaluated in terms of the impact of diferent characteristics of cutting tips and variable spindle speeds.

  7. cDNA cloning, chromosome mapping and expression characterization of human geranylgeranyl pyrophosphate synthase

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Geranylgeranyl pyrophosphate (GGPP) mainly participates in post-translational modification for various proteins including Rho/Rac, Rap and Rab families, as well as in regulation for cell apoptosis. Geranylgeranyl pyrophosphate synthase (GGPPS), which catalyzes the condensation reaction between farnesyl diphosphate and isopentenyl diphosphate, is the key enzyme for synthesizing GGPP. We report the isolation of a gene transcript showing high homology with Drosophila GGPPS cDNA. The transcript is 1 466 bp in length and contains an intact open reading frame (ORF) ranging from nt 239 to 1 138. This ORF encodes a deduced protein of 300 residues with calculated molecular weight of 35 ku. The deduced protein shows 57.5% identity and 75% similarity with Drosophila GGPPS, and contains five characteristic domains of prenyltransferases. Northern hybridization revealed that human GGPPS was expressed highest in heart, and moderately in spleen, testis, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. No obvious bands were detected in other examined tissues. The GGPPS gene was located on human chromosome 1q43 by Radiation Hybrid mapping method. It was proved that there was a putative predisposing gene for prostate cancer in this region, and that analogs of GGPP can inhibit the geranylgeranylation of p21rap protein in PC-3 prostate cancer cell lines. These facts suggest that GGPPS may be one of the candidate genes for prostate cancer.

  8. [Molecular cloning and analysis of cDNA sequences encoding serine proteinase and Kunitz type inhibitor in venom gland of Vipera nikolskii viper].

    Science.gov (United States)

    Ramazanova, A S; Fil'kin, S Iu; Starkov, V G; Utkin, Iu N

    2011-01-01

    Serine proteinases and Kunitz type inhibitors are widely represented in venoms of snakes from different genera. During the study of the venoms from snakes inhabiting Russia we have cloned cDNAs encoding new proteins belonging to these protein families. Thus, a new serine proteinase called nikobin was identified in the venom gland of Vipera nikolskii viper. By amino acid sequence deduced from the cDNA sequence, nikobin differs from serine proteinases identified in other snake species. Nikobin amino acid sequence contains 15 unique substitutions. This is the first serine proteinase of viper from Vipera genus for which a complete amino acid sequence established. The cDNA encoding Kunitz type inhibitor was also cloned. The deduced amino acid sequence of inhibitor is homologous to those of other proteins from that snakes of Vipera genus. However there are several unusual amino acid substitutions that might result in the change of biological activity of inhibitor.

  9. Molecular cloning of cDNA for the human tumor-associated antigen CO-029 and identification of related transmembrane antigens

    Energy Technology Data Exchange (ETDEWEB)

    Szala, S.; Kasai, Yasushi; Steplewski, Z.; Rodeck, U.; Koprowski, H.; Linnenbach, A.J. (Wistar Inst. of Anatomy and Biology, Philadelphia, PA (USA))

    1990-09-01

    The human tumor-associated antigen CO-029 is a monoclonal antibody-defined cell surface glycoprotein of 27-34 kDa. By using the high-efficiency COS cell expression system, a full-length cDNA clone for CO-029 was isolated. When transiently expressed in COS cells, the cDNA clone directed the synthesis of an antigen reactive to monoclonal antibody CO-029 in mixed hemadsorption and immunoblot assays. Sequence analysis revealed that CO-029 belongs to a family of cell surface antigens that includes the melanoma-associated antigen ME491, the leukocyte cell surface antigen CD37, and the Sm23 antigen of the parasitic helminth Schistosoma mansoni. CO-029 and ME491 antigen expression and the effect of their corresponding monoclonal antibodies on cell growth were compared in human tumor cell lines of various histologic origins.

  10. Clinically lean; "cutting the crap".

    Science.gov (United States)

    Caldwell, G

    2012-01-01

    Proponents of Lean Philosophy believe that successful businesses must reduce waste in working time and resources to a minimum, and maximise their use in productive work. The productive work of the Acute Medical Unit is to provide effective clinical management to a daily cohort of acutely ill patients. Many Clinicians are cynical about Lean. In this article, Dr Caldwell discusses how many clinicians complain of too much crap in the workplace, which gets in the way of swift, safe high quality clinical care. He argues that "Cutting the Crap" in the Acute Medical Unit is entirely consistent with Lean approaches to management of complex systems.

  11. Cloning and analysis of a cDNA encoding acetohydroxy acid isomeroreductase from G2 pea

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GA3 application. Screening a G2 pea cDNA library using this fragment as a probe, we obtained a 2036 bp full-length cDNA. It contains a 1746 bp open reading frame and encodes a protein of 581 amino acids with a theoretical molecular weight of 64 ku. It shares high-level sequence identity with AAIR genes from other plant species. This cDNA was cloned into expression vector and recombinant E. coli DH5α cells with remarkable AAIR enzyme activity were obtained.

  12. Cloning and bioinformatics analysis of cDNA encoding cattle Smad4 gene

    Institute of Scientific and Technical Information of China (English)

    Xiaohui ZHANG; Shangzhong XU; Xue GAO; Hongyan REN; Jinbao CHEN

    2008-01-01

    The cDNA of cattle Smad4 gene was cloned by RT-PCR, 3' RACE and 5' RACE and got a 3503-bp full-long cDNA sequence. The cloned cattle Smad4 cDNA sequence had been send to GenBank and got an accession number: DQ494856. Cattle Smad4 gene consists of 12 exons and codes 553 amino acids. Cattle Smad4 cDNA shares 99%, 96%, 95%, 91% and 91% similarity in nucleic acid sequences, and 99%, 98%, 98%, 99% and 98% sim-ilarity in amino acid sequences with sheep, pig, human, rat and mouse, respectively. Smad4 cDNA was found in the testes, pancreas, liver, small intestine, ovary, lymph, car-diac muscle, skeleton muscle and thymus gland, which indicated that Smad4 was broadly expressed in cattle.

  13. Molecular cloning and mammalian expression of human beta 2-glycoprotein I cDNA

    DEFF Research Database (Denmark)

    Kristensen, Torsten; Schousboe, Inger; Boel, Espen

    1991-01-01

    Human β2-glycoprotein (β2gpI) cDNA was isolated from a liver cDNA library and sequenced. The cDNA encoded a 19-residue hydrophobic signal peptide followed by the mature β2gpI of 326 amino acid residues. In liver and in the hepatoma cell line HepG2 there are two mRNA species of about 1.4 and 4.3 k......, respectively, hybridizing specifically with the β2gpI cDNA. Upon isoelectric focusing, recombinant β2gpI obtained from expression of β2gpI cDNA in baby hamster kidney cells showed the same pattern of bands as β2gpI isolated from plasma, and at least 5 polypeptides were visible...

  14. Rooting of stem cuttings of ixora

    Directory of Open Access Journals (Sweden)

    Aline De Souza Silva

    2015-08-01

    Full Text Available The ixora is ornamental plant widely used in landscaping. In order to maximize the propagation of cuts, we evaluated the concentrations of auxin (indolbutiric acid and the presence of leaves on the rooting in cuts of Ixora coccinea L. The experiment was conducted in randomized block design, in factorial design 3x4, with three types of cuts (without leaf, with two or four leaves, four concentrations of indolbutiric acid (0, 1000, 2000 and 4000 mg L-1, with four replications and 10 cuts in each experimental unit. After 53 days of implantation the experiment, evaluated the survival(%, rooting(%, sprouting(%, formation of callus(%, number, length and biomass of roots formed. The interaction of the type of cuts with concentrations of auxin was not significant for any of the variables analyzed. The survival of cuttings was not influenced by the treatments. Cuts with two or four leaves presented rooting and length of roots above the cuttings without leaves. The application of auxin does not substitute the presence of leaf in cuts of ixora in vegetative propagation. The vegetative propagation by cut of ixora can be made without application of auxin, and the leaves must be maintained in the cuttings.

  15. The ongoing violence against women: Female Genital Mutilation/Cutting.

    Science.gov (United States)

    Muteshi, Jacinta K; Miller, Suellen; Belizán, José M

    2016-04-18

    Female Genital Mutilation/Cutting (FGM/C) comprises different practices involving cutting, pricking, removing and sometimes sewing up external female genitalia for non-medical reasons. The practice of FGM/C is highly concentrated in a band of African countries from the Atlantic coast to the Horn of Africa, in areas of the Middle East such as Iraq and Yemen, and in some countries in Asia like Indonesia. Girls exposed to FGM/C are at risk of immediate physical consequences such as severe pain, bleeding, and shock, difficulty in passing urine and faeces, and sepsis. Long-term consequences can include chronic pain and infections. FGM/C is a deeply entrenched social norm, perpetrated by families for a variety of reasons, but the results are harmful. FGM/C is a human rights issue that affects girls and women worldwide. The practice is decreasing, due to intensive advocacy activities of international, national, and grassroots agencies. An adolescent girl today is about a third less likely to be cut than 30 years ago. However, the rates of abandonment are not high enough, and change is not happening as rapidly as necessary. Multiple interventions have been implemented, but the evidence base on what works is lacking. We in reproductive health must work harder to find strategies to help communities and families abandon these harmful practices.

  16. Completion sequence and cloning of the infectious cDNA of a chb isolate of cucumber green mottle mosaic virus.

    Science.gov (United States)

    Zhong, M; Zhao, X; Liu, Y; Wang, Y; Cao, K

    2015-03-01

    Cucumber green mottle mosaic virus (CGMMV) is an important and widespread seed-borne virus that infects Cucurbitaceous plants. It is a member of the genus Tobamovirus in the family Virgaviridae with a monopartite (+) ssRNA genome. Here we report the complete genome sequence, construction and testing of the infectious clones of a chb isolate of CGMMV. Full-length CGMMV cDNA was cloned into the vector pUC19. The linearized vector containing full-length cDNA was used as template for in vitro transcription, and the synthesized capped transcript was highly infectious in Chenopodium amaranticolor and cucumber (Cucumis sativus). Inoculated plants showed symptoms typical of CGMMV infection. The infectivity was confirmed by mechanical transmission to new plants, RT-PCR and western blot. Progeny virus derived from infectious transcripts had the same biological and biochemical properties as wild-type virus. To our knowledge, this is the first detailed report of a biologically active transcript from CGMMV.

  17. Purification, cDNA cloning, and characterization of LysM-containing plant chitinase from horsetail (Equisetum arvense).

    Science.gov (United States)

    Inamine, Saki; Onaga, Shoko; Ohnuma, Takayuki; Fukamizo, Tamo; Taira, Toki

    2015-01-01

    Chitinase-A (EaChiA), molecular mass 36 kDa, was purified from the vegetative stems of a horsetail (Equisetum arvense) using a series of column chromatography. The N-terminal amino acid sequence of EaChiA was similar to the lysin motif (LysM). A cDNA encoding EaChiA was cloned by rapid amplification of cDNA ends and polymerase chain reaction. It consisted of 1320 nucleotides and encoded an open reading frame of 361 amino acid residues. The deduced amino acid sequence indicated that EaChiA is composed of a N-terminal LysM domain and a C-terminal plant class IIIb chitinase catalytic domain, belonging to the glycoside hydrolase family 18, linked by proline-rich regions. EaChiA has strong chitin-binding activity, however, no antifungal activity. This is the first report of a chitinase from Equisetopsida, a class of fern plants, and the second report of a LysM-containing chitinase from a plant.

  18. Characterization and expression of a cDNA encoding a tubuliform silk protein of the golden web spider Nephila antipodiana.

    Science.gov (United States)

    Huang, W; Lin, Z; Sin, Y M; Li, D; Gong, Z; Yang, D

    2006-07-01

    Spider silks are renowned for their excellent mechanical properties. Although several spider fibroin genes, mainly from dragline and capture silks, have been identified, there are still many members in the spider fibroin gene family remain uncharacterized. In this study, a novel silk cDNA clone from the golden web spider Nephila antipodiana was isolated. It is serine rich and contains two almost identical fragments with one varied gap region and one conserved spider fibroin-like C-terminal domain. Both in situ hybridization and immunoblot analyses have shown that it is specifically expressed in the tubuliform gland. Thus, it likely encodes the silk fibroin from the tubuliform gland, which supplies the main component of the inner egg case. Unlike other silk proteins, the protein encoded by the novel cDNA in water solution exhibits the characteristic of an alpha-helical protein, which implies the distinct property of the egg case silk, though the fiber of tubuliform silk is mainly composed of beta-sheet structure. Its sequence information facilitates elucidation of the evolutionary history of the araneoid fibroin genes.

  19. Isolation and expression of the full-length cDNA encoding CD59 antigen of human lymphocytes.

    Science.gov (United States)

    Sawada, R; Ohashi, K; Anaguchi, H; Okazaki, H; Hattori, M; Minato, N; Naruto, M

    1990-04-01

    To identify the primary structure of CD59 antigen and to elucidate its function, a full-length cDNA clone of CD59 was isolated. The cDNA sequence contained an open reading frame that encodes an 128-amino-acid peptide. The amino-terminal 25 amino acids represented a typical signal peptide sequence and the carboxy-terminal hydrophobic amino acids were characteristic for phosphatidylinositol-anchored proteins. The predicted mature protein sequence showed 35% homology with murine Ly-6C.1 and 31% with Ly-6A.2. The number and the distribution of cysteine residues were conserved, implying that the CD59 represented a human homologue of murine Ly-6. RNA blot hybridization analysis revealed the expression of CD59 mRNA in placental, lung, and pancreatic tissues. The mRNA was not only expressed in T-cell lines but in some of monocytic, myeloid, and B-cell lines. In all of these tissues and cell lines, at least four mRNA species were detected. DNA blot hybridization analysis revealed a rather simple genomic structure, which suggested a single gene as compared with the complex multigene family of murine Ly-6.

  20. Meat speciation by restriction fragment length polymorphism analysis using an α-actin cDNA probe.

    Science.gov (United States)

    Fairbrother, K S; Hopwood, A J; Lockley, A K; Bardsley, R G

    1998-09-01

    Classical DNA fingerprinting is based on separation of DNA restriction fragments by electrophoresis and hybridisation to nucleic acid probes containing repetitive nucleotide sequences. The use of such mini- or micro-satellite probes tends to yield patterns specific to an individual rather than to a species, hence their value in forensic analysis but general unsuitability for meat speciation. In the present study, a cDNA probe based on conserved sequences contained in members of the actin multigene family has been evaluated for potential application in meat speciation. Genomic DNA was extracted from muscle and digested with BamHI before electrophoresis and hybridisation to a murine α-actin cDNA probe. Beef, pork, lamb, horse, chicken and fish DNA restriction fragments formed characteristic 'fingerprints' which were reproducible and varied sufficiently to allow discrimination even between closely-related species. However no major differences were seen between individuals of the same breed or between different breeds within a species. When DNA obtained from fresh tissue and also from meat heated at 120 °C was analysed, the gel patterns were essentially the same. An attractive feature of this approach is that it employs a single cross-reacting probe and set of conditions, and gives different patterns with all species so far studied. This simplicity suggests applications in meat speciation or related areas of biology.

  1. Cloning and characterization of a cDNA encoding type 1 diacylglycerol acyltransferase from sunflower (Helianthus annuus L.).

    Science.gov (United States)

    Sun, Li; Ouyang, Chao; Kou, Shanglong; Wang, Shenghua; Yao, Yunyi; Peng, Tong; Xu, Ying; Tang, Lin; Chen, Fang

    2011-01-01

    A full-length cDNA encoding a putative diacylglycerol acyltransferase (DGAT; EC 2.3.1.20) was obtained from sunflower (Helianthus annuus L.) seeds. The 1524-bp open reading frame of this cDNA, designated as HaDGAT1, encodes a protein of 507 amino acids with a molecular mass of 58.5 kDa showing high homology to DGAT1 enzymes of other plants. The protein characters, such as a predicted structure with a long N-terminal hydrophilic domain followed by 9 transmembrane domains, acyl-CoA-binding signature, diacylglycerol (DAG)-binding and putative endoplasmic reticulum retrieval motifs (ER-DIR), also indicated that HaDGAT belongs to the DGAT1 family. HaDGAT1 is expressed in all plant tissues especially in developing seeds. Expression of recombinant HaDGAT1 in yeast showed an 1.76-fold increase of total fatty acids, especially unsaturated fatty acids such as palmitoleic acid (enhanced by 86.6%) and oleic acid (enhanced by 81.6%).

  2. The center-cut solution.

    Science.gov (United States)

    Firnstahl, T W

    1993-01-01

    Timothy Firnstahl owns five successful restaurants in Seattle, but he recently came very close to owning none. In the early 1990s, he found himself, like so many restauranteurs, facing rising costs, inefficient management, and a recession. Confronting financial annihilation, Firnstahl had to act quickly: since he had no peripherals to trim, he cut off the head of his company. Remarkably, it worked. Firnstahl's problem was his new and innovative restaurant, Sharps Fresh Roasting. The heart of the Sharps concept was a unique long-roasting technique that made lean, inexpensive meats taste as juicy and delicious as fattier, expensive cuts. The process also lent itself to faster service and lower labor costs. But it wasn't working. Sharps wasn't breaking even, and his other restaurants couldn't make up the difference. He needed a solution fast. Firnstahl got his answer from Mikhail Gorbachev: slash the centralized command and liberate the company. In doing so, he would also transfer virtually all power and responsibility to his line managers. And after five months of intensive study and planning, he accomplished what he set out to do. He fired most of his corporate staff, empowered his restaurant managers with "100% Power and Responsibility," and, finally, undertook a massive promotion campaign. A year later, Sharps Fresh Roasting is the gold mine Firnstahl always believed it could be. He's done away with bureaucracy and turned business around in a down market. All this because his managers are managing themselves.

  3. CDNA library from the Latex of Hevea brasiliensis

    Directory of Open Access Journals (Sweden)

    Wilaiwan Chotigeat

    2010-12-01

    Full Text Available Latex from Hevea brasiliensis contains 30-50% (w/w of natural rubber (cis-1,4-polyisoprene, the important rawmaterial for many rubber industries. We have constructed a cDNA library from the latex of H. brasiliensis to investigate theexpressed genes and molecular events in the latex. We analyzed 412 expressed sequence tags (ESTs. More than 90% of theEST clones showed homology to previously described sequences in public databases. Functional classification of the ESTsshowed that the largest category were proteins of unknown function (30.1%, 11.4% of ESTs encoded for rubber synthesisrelatedproteins (RS and 8.5% for defense or stress related proteins (DS. Those with no significant homology to knownsequences (NSH accounted for 8.7%, primary metabolism (PM and gene expression and RNA metabolism were 7.8% and6.6%, respectively. Other categories included, protein synthesis-related proteins (6.6%, chromatin and DNA metabolism(CDM 3.9%, energy metabolism (EM 3.4%, cellular transport (CT 3.2%, cell structure (CS 3.2%, signal transduction (ST2.2%, secondary metabolism (SM 1.7%, protein fate (PF 2.2%, and reproductive proteins (RP 0.7%.

  4. Effects of heat on cut mark characteristics.

    Science.gov (United States)

    Waltenberger, Lukas; Schutkowski, Holger

    2017-02-01

    Cut marks on bones provide crucial information about tools used and their mode of application, both in archaeological and forensic contexts. Despite a substantial amount of research on cut mark analysis and the influence of fire on bones (shrinkage, fracture pattern, recrystallisation), there is still a lack of knowledge in cut mark analysis on burnt remains. This study provides information about heat alteration of cut marks and whether consistent features can be observed that allow direct interpretation of the implemented tools used. In a controlled experiment, cut marks (n=25) were inflicted on pig ribs (n=7) with a kitchen knife and examined using micro-CT and digital microscopy. The methods were compared in terms of their efficacy in recording cut marks on native and heat-treated bones. Statistical analysis demonstrates that floor angles and the maximum slope height of cuts undergo significant alteration, whereas width, depth, floor radius, slope, and opening angle remain stable. Micro-CT and digital microscopy are both suitable methods for cut mark analysis. However, significant differences in measurements were detected between both methods, as micro-CT is less accurate due to the lower resolution. Moreover, stabbing led to micro-fissures surrounding the cuts, which might also influence the alteration of cut marks.

  5. Screening of genes for proteins interacting with the PS1TP5 protein of hepatitis B virus: probing a human leukocyte cDNA library using the yeast two-hybrid system

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jian-kang; ZHAO Long-feng; CHENG Jun; GUO Jiang; LUN Yong-zhi; HONG Yuan

    2006-01-01

    Background The hepatitis B virus (HBV) genome includes S, C, P and X regions. The S region is divided into four subregions of pre-pre-S, pre-S1, pre-S2 and S. PS1TP5 (human gene 5 transactivated by pre-S1 protein of HBV) is a novel target gene transactivated by the pre-S1 protein that has been screened with a suppression subtractive hybridization technique in our laboratory (GenBank accession: AY427953). In order to investigate the biological function of the PS1TP5 protein, we performed a yeast two-hybrid system 3 to screen proteins from a human leukocyte cDNA library interacting with the PS 1TP5 protein.Methods The reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify the gene of PS1TP5 from the mRNA of HepG2 cells and the gene was then cloned into the pGEM-T vector. After being sequenced and analyzed with Vector NTI 9.1 and NCBI BLAST software, the target gene of PS1TP5 was cut from the pGEM-T vector and cloned into a yeast expression plasmid pGBKT7, then "bait" plasmid pGBKT7-PS 1TP5 was transformed into the yeast strain AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting hybridization.After expression of the pGBKT7-PS1TP5 fusion protein in the AH109 yeast strain was accomplished, a yeast two-hybrid screening was performed by mating AH 109 with Y 187 containing a leukocyte cDNA library plasmid.The mated yeast was plated on quadruple dropout medium and assayed for α-gal activity. The interaction between the PS1TP5 protein and the proteins obtained from positive colonies was further confirmed by repeating the yeast two-hybrid screen. After extracting and sequencing of plasmids from blue colonies we carried out a bioinformatic analysis.Results Forty true positive colonies were selected and sequenced, full length sequences were obtained and we searched for homologous DNA sequences from GenBank. Among the 40 positive colonies, 23 coding genes

  6. Construction and analysis of SSH cDNA library of human vascular endothelial cells related to gastrocarcinoma

    OpenAIRE

    2003-01-01

    AIM: To construct subtracted cDNA libraries of human vascular endothelial cells (VECs) related to gastrocarcinoma using suppression substractive hybridization (SSH) and to analyze cDNA libraries of gastrocarcinoma and VECs in Cancer Gene Anatomy Project (CGAP) database.

  7. Random rapid amplification of cDNA ends (RRACE) allows for cloning of multiple novel human cDNA fragments containing (CAG)n repeats.

    Science.gov (United States)

    Carney, J P; McKnight, C; VanEpps, S; Kelley, M R

    1995-04-03

    We describe a new technique for isolating cDNA fragments in which (i) either a partial sequence of the cDNA is known or (ii) a repeat sequence is utilized. We have used this technique, termed random rapid amplification of cDNA ends (random RACE), to isolate a number of trinucleotide repeat (CAG)n-containing genes. Using the random RACE (RRACE) technique, we have isolated over a hundred (CAG)n-containing genes. The results of our initial analysis of ten clones indicate that three are identical to previously cloned (CAG)n-containing genes. Three of our clones matched with expressed sequence tags, one of which contained a CA repeat. The remaining four clones did not match with any sequence in GenBank. These results indicate that this approach provides a rapid and efficient method for isolating trinucleotide repeat-containing cDNA fragments. Finally, this technique may be used for purposes other than cloning repeat-containing cDNA fragments. If only a partial sequence of a gene is known, our system, described here, provides a rapid and efficient method for isolating a fragment of the gene of interest.

  8. Isolation of cDNA clones coding for human tissue factor: primary structure of the protein and cDNA

    Energy Technology Data Exchange (ETDEWEB)

    Spicer, E.K.; Horton, R.; Bloem, L.; Bach, R.; Williams, K.R.; Guha, A.; Kraus, J.; Lin, T.C.; Nemerson, Y.; Konigsberg, W.H.

    1987-08-01

    Tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor VII and calcium. lambda Phage containing the tissue factor gene were isolated from a human placental cDNA library. The amino acid sequence deduced from the nucleotide sequence of the cDNAs indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptide chain composed of 263 residues. The derived primary structure of tissue factor has been confirmed by comparison to protein and peptide sequence data. The sequence of the mature protein suggests that there are three distinct domains: extracellular, residues 1-219; hydrophobic, residues 220-242; and cytoplasmic, residues 243-263. Three potential N-linked carbohydrate attachment sites occur in the extracellular domain. The amino acid sequence of tissue factor shows no significant homology with the vitamin K-dependent serine proteases, coagulation cofactors, or any other protein in the National Biomedical Research Foundation sequence data bank (Washington, DC).

  9. cDNA amplification by SMART-PCR and suppression subtractive hybridization (SSH)-PCR.

    Science.gov (United States)

    Hillmann, Andrew; Dunne, Eimear; Kenny, Dermot

    2009-01-01

    The comparison of two RNA populations that differ from the effects of a single-independent variable, such as a drug treatment or a specific genetic defect, can identify differences in the abundance of specific transcripts that vary in a population-dependent manner. There are a variety of methods for identifying differentially expressed genes, including microarray, SAGE, qRT-PCR, and DDGE. This protocol describes a potentially less sensitive yet relatively easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under investigation and is particularly applicable when minimal levels of starting material, RNA, are available. RNA input can often be a limiting factor when analyzing RNA from, for example, rigorously purified blood cells. This protocol describes the use of SMART-PCR to amplify cDNA from sub-microgram levels of RNA. The amplified cDNA populations under comparison are then subjected to suppression subtractive hybridization (SSH-PCR), a technique that couples subtractive hybridization with suppression PCR to selectively amplify fragments of differentially expressed genes. The final products are cDNA populations enriched for significantly over-represented transcripts in either of the two input RNA preparations. These cDNA populations may then be cloned to make subtracted cDNA libraries and/or used as probes to screen subtracted cDNA, global cDNA, or genomic DNA libraries.

  10. RICD: A rice indica cDNA database resource for rice functional genomics

    Directory of Open Access Journals (Sweden)

    Zhang Qifa

    2008-11-01

    Full Text Available Abstract Background The Oryza sativa L. indica subspecies is the most widely cultivated rice. During the last few years, we have collected over 20,000 putative full-length cDNAs and over 40,000 ESTs isolated from various cDNA libraries of two indica varieties Guangluai 4 and Minghui 63. A database of the rice indica cDNAs was therefore built to provide a comprehensive web data source for searching and retrieving the indica cDNA clones. Results Rice Indica cDNA Database (RICD is an online MySQL-PHP driven database with a user-friendly web interface. It allows investigators to query the cDNA clones by keyword, genome position, nucleotide or protein sequence, and putative function. It also provides a series of information, including sequences, protein domain annotations, similarity search results, SNPs and InDels information, and hyperlinks to gene annotation in both The Rice Annotation Project Database (RAP-DB and The TIGR Rice Genome Annotation Resource, expression atlas in RiceGE and variation report in Gramene of each cDNA. Conclusion The online rice indica cDNA database provides cDNA resource with comprehensive information to researchers for functional analysis of indica subspecies and for comparative genomics. The RICD database is available through our website http://www.ncgr.ac.cn/ricd.

  11. [Construction and analysis of subtractive cDNA library associated with multidrug resistance of acute leukemia].

    Science.gov (United States)

    Ji, Lei; Zhang, Wang-Gang; Liu, Jie; Liu, Xin-Ping; Yao, Li-Bo

    2004-08-01

    The study was aimed to construct subtractive cDNA library associated with multidrug resistance (MDR) of acute leukemia for screening genes related to MDR in leukemia. The improved PCR-based subtractive hybridization was performed to clone differential genes between HL-60/VCR and HL-60 cell line. The mRNA of HL-60/VCR and HL-60 cell line were isolated. Then the mRNA of HL-60/VCR group was reversely transcribed into cDNA by Cap-Finder method, and the mRNA of HL-60 was reversely transcribed into cDNA by ordinary method to be marked by biotin for the hybridization next with HL-60/VCR cDNA. After hybridizing, filtrating through the sephacryl S-400 column, absorbing by the magnetic beads, and amplifying by PCR method, the fragments were cloned by T-A method and the cDNA library was constructed. Then the quality of cDNA library was identified by dot-blotting hybridization method. The results showed that after constriction, the library demonstrated its good quality. There was a high proportion of large fragments in this library. From small amount of samples a large amount of candidate fragments could be screened rapidly at once by dot-blotting hybridization. It is concluded that a differentially-expressed subtractive cDNA library in MDR of leukemia with high quality and larger fragments can be efficiently constructed by improving subtractive hybridization and selective PCR method.

  12. Construction of full length cDNA expression library of hepatopancreas of Penaeus monodon

    Institute of Scientific and Technical Information of China (English)

    罗田; 徐洵

    2002-01-01

    --mRNA was isolated from the hepatopancrease of shrimp Penaeus monodon with a PolyATtract System 1000 Kit. By using mRNA as template, double- strand cDNA with EcoR I/Xho I ends was synthesized by using a ZAP Express cDNA Synthesis Kit. The cDNA was inserted into the lambda ZAP Express vector predigested with EcoR I/Xho I, and the recombinant DNA was in vitro packaged into larnbda phage with GigapackⅢ Gold packaging extracts. These recombinant phages were then used to transfect E. coli XLl - Blue MRF', and finally a cDNA expression library was constructed. The library is 7.2 × 105pfu in capacity and its recombination ratio is higher than 99%. The size of the inserted cDNAs was determined by EcoR I/Xho I digestion of 9 phagemids prepared by in vivo excision of plaques selected randomly from amplified cDNA library . The longest inserted cDNA is about 1.6 kb in length. The complete sequence (about 1.2 kb) of actin cDNA was amplified from the library by PCR reveals that this library contains full-length cDNAs of Penaeus mod on hepatopancreas and is available for screening and expression of shrimp genes.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1991-02-26

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

  14. Cloning of lea cDNA fragment of carrot (Daucus carota L.) and analysis of its expression features

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Addition of concentrated sucrose to MS culture arrests the development of carrot somatic embryo at the stage of cotyledon embryo and, with the sucrose concentration restored to normal level, the embryo thus arrested is reactivated into post-embryonic development. Using the method of RT-PCR, the cDNA fragment of a new member of the Dc3 family of lea has been obtained from carrot somatic embryo under regulated state. As revealed by Northern blotting, strong expression has been observed in carrot somatic embryo under regulated state but the expression was much reduced 12 h after deregulation, and nearly disappeared 24 h after. Based on this finding as well as results of related studies, it is surmised that changing the sucrose concentration in culture enabled carrot somatic embryo under suspension culture to undergo a specific course of development which is comparable to the dormancy-germination process of seeds.

  15. Circular rapid amplification of cDNA ends for high-throughput extension cloning of partial genes.

    Science.gov (United States)

    Fu, Glenn K; Wang, Jonathan T; Yang, Junming; Au-Young, Janice; Stuve, Laura L

    2004-07-01

    The rapid amplification of cDNA ends (RACE) procedure is a widely used PCR-based method to clone the cDNA ends of mRNA transcripts. Current RACE methods often produce a high background of nonspecific PCR products, which can exclude the identification of the target cDNA of interest. We describe here an improved RACE procedure using circular cDNA templates and demonstrate the successful extension cloning of 4406 cDNAs.

  16. Testing of Commercial Cutting Heads for Abrasive Water Jet Technology

    OpenAIRE

    Klich, J. (Jiří); Hlaváček, P.; M. Zeleňák; Sitek, L. (Libor); Foldyna, J. (Josef)

    2013-01-01

    Five different cutting heads designed for cutting by high-speed abrasive water jet technology were tested from cutting ability point of view. Straight kerfs were cut in several metal materials by abrasive water jet. Material removal volume was determined as a measure of performance of specific cutting head. Quality of cutting surface was observed, too. Results are compared and discussed.

  17. Construction of cDNA subtractive library from pearl oyster ( Pinctada fucata Gould) with red color shell by SSH

    Science.gov (United States)

    Guan, Yunyan; Huang, Liangmin; He, Maoxian

    2011-05-01

    The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively. Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6, shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COI. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COI so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster.

  18. Construction of cDNA subtractive library from pearl oyster (Pinctada fucata Gould) with red color shell by SSH

    Institute of Scientific and Technical Information of China (English)

    GUAN Yunyan; HUANG Liangmin; HE Maoxian

    2011-01-01

    The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively.Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6,shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COI. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COI so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster.

  19. Cloning and sequencing of dolphinfish (Coryphaena hippurus, Coryphaenidae) growth hormone-encoding cDNA.

    Science.gov (United States)

    Peduel, A D; Elizur, A; Knibb, W

    1994-01-01

    The cDNA encoding the preprotein growth hormone from the dolphinfish (Coryphaena hippurus) has been cloned and sequenced. The cDNA was derived by reverse transcription of RNA from the pituitary of a young fish using the method known as Rapid Amplification of cDNA Ends (RACE). An oligonucleotide primer corresponding to the 5' region of Pagrus major and the universal RACE primer enabled amplification using the Polymerase Chain Reaction (PCR). The dolphinfish and yellow-tail, Seriola quineqeradiata, are both members of the sub-order Percoidei (Perciforme) and their GH sequences show a high level of homology.

  20. Characterization of the human HOX 7 cDNA and identification of polymorphic markers.

    Science.gov (United States)

    Padanilam, B J; Stadler, H S; Mills, K A; McLeod, L B; Solursh, M; Lee, B; Ramirez, F; Buetow, K H; Murray, J C

    1992-09-01

    cDNA clones for a human HOX 7 gene obtained with homologous clones of Drosophila were used in human gene mapping studies. The human cDNA clone was isolated from a library constructed from human embryonic craniofacial material. The sequence of the cDNA demonstrates significant homology with mouse HOX 7. A search for RFLPs identified MboII and BstEII variants. A CA dinucleotide repeat with 5 alleles was also identified and allowed placement of HOX 7 into a defined linkage map. Evidence for linkage disequilibrium was found with markers tested. These results place the human HOX 7 gene in a defined position on 4p.

  1. Characterization and simulation of cDNA microarray spots using a novel mathematical model

    OpenAIRE

    2007-01-01

    Abstract Background The quality of cDNA microarray data is crucial for expanding its application to other research areas, such as the study of gene regulatory networks. Despite the fact that a number of algorithms have been suggested to increase the accuracy of microarray gene expression data, it is necessary to obtain reliable microarray images by improving wet-lab experiments. As the first step of a cDNA microarray experiment, spotting cDNA probes is critical to determining the quality of s...

  2. A CLINICAL STUDY ON PROSTATE CANCER DIAGNOSIS WITH cDNA MACROARRAY

    Institute of Scientific and Technical Information of China (English)

    ZHONG Wei-de; XIE Ke-ji; WEI Hong-ai; LIU Liang-shi; LIU Wen-hua; JIANG Fu-neng; ZENG Guang-qiao; DAI Qi-shan; HE Hui-chan; BI Xue-cheng; PENG Zhi-qiang

    2005-01-01

    Objective: To diagnose prostatic cancer (CaP) with cDNA macroarray. Methods: Total RNA was isolated from patients with prostate cancer and from normal people, and poly(A) RNA was further purified. Then differentially expressed genes were analysed in CaP and normal prostate by cDNA macroarray system. Results: There were differential expressions of nine prostate-associated specific genes in CaP as compared with normal prostate, among which, 7 were significantly up-regulated and 2 were down-regulated. Conclusion: As a diagnostic approach at molecular level, the cDNA macroarray is supposed to elevate the detection rate of CaP.

  3. Ceramic cutting tools materials, development and performance

    CERN Document Server

    Whitney, E Dow

    1994-01-01

    Interest in ceramics as a high speed cutting tool material is based primarily on favorable material properties. As a class of materials, ceramics possess high melting points, excellent hardness and good wear resistance. Unlike most metals, hardness levels in ceramics generally remain high at elevated temperatures which means that cutting tip integrity is relatively unaffected at high cutting speeds. Ceramics are also chemically inert against most workmetals.

  4. A cactus theorem for end cuts

    CERN Document Server

    Evangelidou, Anastasia

    2011-01-01

    Dinits-Karzanov-Lomonosov showed that it is possible to encode all minimal edge cuts of a graph by a tree-like structure called a cactus. We show here that minimal edge cuts separating ends of the graph rather than vertices can be `encoded' also by a cactus. We apply our methods to finite graphs as well and we show that several types of cuts can be encoded by cacti.

  5. Machining Challenges: Macro to Micro Cutting

    Science.gov (United States)

    Shunmugam, M. S.

    2016-04-01

    Metal cutting is an important machining operation in the manufacture of almost all engineering components. Cutting technology has undergone several changes with the development of machine tools and cutting tools to meet challenges posed by newer materials, complex shapes, product miniaturization and competitive environments. In this paper, challenges in macro and micro cutting are brought out. Conventional and micro end-milling are included as illustrative examples and details are presented along with discussion. Lengthy equations are avoided to the extent possible, as the emphasis is on the basic concepts.

  6. A cutting force model for micromilling applications

    DEFF Research Database (Denmark)

    Bissacco, Giuliano; Hansen, Hans Nørgaard; De Chiffre, Leonardo

    2006-01-01

    In micro milling the maximum uncut chip thickness is often smaller than the cutting edge radius. This paper introduces a new cutting force model for ball nose micro milling that is capable of taking into account the effect of the edge radius.......In micro milling the maximum uncut chip thickness is often smaller than the cutting edge radius. This paper introduces a new cutting force model for ball nose micro milling that is capable of taking into account the effect of the edge radius....

  7. Cutting food in terrestrial carnivores and herbivores.

    Science.gov (United States)

    Sanson, Gordon

    2016-06-06

    Insects and mammals cut their food up into small pieces to facilitate ingestion and chemical digestion. Teeth and jaws act as cutting tools, but, unlike engineering tools designed for a specific purpose, must generally cope with substantial variation in food properties and work at many scales. Knowing how teeth and jaws work effectively requires an understanding of the cutting on the edges and the mechanisms that remove cut material. Variability and heterogeneity of diet properties are not well known, and, for example, may be higher and overlap more in the browsing and grazing categories of plant diets. A reinterpretation of tooth function in large mammal browsers and grazers is proposed.

  8. Underwater laser cutting of metal structures

    Energy Technology Data Exchange (ETDEWEB)

    Alfille, J.P.; Prunele, D. de [CEA Centre d`Etudes de Saclay, 91 - Gif-sur-Yvette (France). Direction des Technologies Avancees; Pilot, G. [CEA Centre d`Etudes de Fontenay-aux-Roses, 92 (France). Dept. de Protection de l`Environnement et des Installations; Fredrick, P.; Ramaswami, V.S.; Muys, P. [Radius Engineering, Gent (Belgium)

    1994-12-31

    Cutting tests were carried out on stainless steel (304L) in air and under 7 meters of water (application to reactor pools), using CO{sub 2} and YAG lasers; results concerned cutting speed, quality of cut, cutting thickness. By-products of sectioning operations using a CO{sub 2} laser were studied: dross, aerosols, suspended particles in water, gas analysis, chemical analysis of the aerosols. Same measurements are currently being taken in the case of the YAG laser with beam transported via optical fiber. (from author). 16 figs., 2 tabs., 3 refs.

  9. Cutting Properties of Austenitic Stainless Steel by Using Laser Cutting Process without Assist Gas

    Directory of Open Access Journals (Sweden)

    Hitoshi Ozaki

    2012-01-01

    Full Text Available Recently, laser cutting is used in many industries. Generally, in laser cutting of metallic materials, suitable assist gas and its nozzle are needed to remove the molten metal. However, because of the gas nozzle should be set closer to the surface of a workpiece, existence of the nozzle seems to prevent laser cutting from being used flexible. Therefore, the new cutting process, Assist Gas Free laser cutting or AGF laser cutting, has been developed. In this process, the pressure at the bottom side of a workpiece is reduced by a vacuum pump, and the molten metal can be removed by the air flow caused by the pressure difference between both sides of the specimen. In this study, cutting properties of austenitic stainless steel by using AGF laser cutting with 2 kW CO2 laser were investigated. Laser power and cutting speed were varied in order to study the effect of these parameters on cutting properties. As a result, austenitic stainless steel could be cut with dross-free by AGF laser cutting. When laser power was 2.0 kW, cutting speed could be increased up to 100 mm/s, and kerf width at specimen surface was 0.28 mm.

  10. 5'-end sequences of budding yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us Budding yeast cDNA sequencing project 5'-end sequences of budding yeast full-length cDNA clones and quality score...s Data detail Data name 5'-end sequences of budding yeast full-length cDNA clones and quality score...or-capping method, the sequence quality score generated by the Phred software, and links to SGD, dbEST and U...es. FASTA format. Quality Phred's quality score About This Database Database Desc...g yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive ...

  11. cDNA cloning of the murine PEX gene implicated in X-linked hypophosphatemia and evidence for expression in bone

    Energy Technology Data Exchange (ETDEWEB)

    Du, L.; Desbarats, M.; Viel, J. [McGill Univ., Montreal, Quebec (Canada)] [and others

    1996-08-15

    The recently identified human PEX g ene apparently encodes for a neutral endopeptidase that is mutated in patients with X-linked hypophosphatemia. The 3{prime} and 5{prime} ends of the coding region of PEX have not been cloned, nor has the tissue expression of the gene been identified. Here we report the isolation and characterization of the complete open reading frame of the mouse Pex gene and the demonstration of its expression in bone. Mouse Pex cDNA is predicted to encode a protein of 749 amino acids with 95% identity to the available human PEX sequence and significant homology to members of the membrane-bound metalloendopeptidase family. Northern blot analysis revealed a 6.6-kb transcript in bone and in cultured osteoblasts from normal mice that was not detectable in samples from the Hyp mouse, the murine homolog of human X-linked hypophosphatemia. Pex transcripts were, however, detectable in Hyp bone by RT-PCR amplification. Of particular interest, a cDNA clone from rat incisor shows 93% sequence identity to the 5{prime} end of Pex cDNA, suggesting that Pex may be expressed in another calcified tissue, the tooth. The association of impaired mineralization of bone and teeth and disturbed renal phosphate reabsorption with altered expression of Pex suggests that the Pex gene product may play a critical role in these processes. 47 refs., 2 figs., 1 tab.

  12. Characterization of Leukemia-Inducing Genes Using a Proto-Oncogene/Homeobox Gene Retroviral Human cDNA Library in a Mouse In Vivo Model.

    Directory of Open Access Journals (Sweden)

    Su Hwa Jang

    Full Text Available The purpose of this research is to develop a method to screen a large number of potential driver mutations of acute myeloid leukemia (AML using a retroviral cDNA library and murine bone marrow transduction-transplantation system. As a proof-of-concept, murine bone marrow (BM cells were transduced with a retroviral cDNA library encoding well-characterized oncogenes and homeobox genes, and the virus-transduced cells were transplanted into lethally irradiated mice. The proto-oncogenes responsible for leukemia initiation were identified by PCR amplification of cDNA inserts from genomic DNA isolated from leukemic cells. In an initial screen of ten leukemic mice, the MYC proto-oncogene was detected in all the leukemic mice. Of ten leukemic mice, 3 (30% had MYC as the only transgene, and seven mice (70% had additional proto-oncogene inserts. We repeated the same experiment after removing MYC-related genes from the library to characterize additional leukemia-inducing gene combinations. Our second screen using the MYC-deleted proto-oncogene library confirmed MEIS1and the HOX family as cooperating oncogenes in leukemia pathogenesis. The model system we introduced in this study will be valuable in functionally screening novel combinations of genes for leukemogenic potential in vivo, and the system will help in the discovery of new targets for leukemia therapy.

  13. Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 (HSC70) in Fenneropenaeus chinensis

    Institute of Scientific and Technical Information of China (English)

    JIAO Chuanzhen; WANG Zaizhao; LI Fuhua; ZHANG Chengsong; XIANG Jianhai

    2004-01-01

    The cDNA encoding hsc70 of Chinese shrimp Fenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5′- and 3′-untranslated region(5′- and 3′-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1-547 bp, but they did not exist in the region of 548-2090 bp of hsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 from Oncorhynchus mykiss and HSC70 from Homo sapiens. It also exhibited 85.8% similarity with HSP70 from Mus musculu and 85.4% with HSC70 from Manduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills in Fenneropenaeus chinensis. No difference could be detected on hsc70 mRNA level in muscle between heat-shocked and control animals.

  14. Characterization of Leukemia-Inducing Genes Using a Proto-Oncogene/Homeobox Gene Retroviral Human cDNA Library in a Mouse In Vivo Model.

    Science.gov (United States)

    Jang, Su Hwa; Lee, Sohyun; Chung, Hee Yong

    2015-01-01

    The purpose of this research is to develop a method to screen a large number of potential driver mutations of acute myeloid leukemia (AML) using a retroviral cDNA library and murine bone marrow transduction-transplantation system. As a proof-of-concept, murine bone marrow (BM) cells were transduced with a retroviral cDNA library encoding well-characterized oncogenes and homeobox genes, and the virus-transduced cells were transplanted into lethally irradiated mice. The proto-oncogenes responsible for leukemia initiation were identified by PCR amplification of cDNA inserts from genomic DNA isolated from leukemic cells. In an initial screen of ten leukemic mice, the MYC proto-oncogene was detected in all the leukemic mice. Of ten leukemic mice, 3 (30%) had MYC as the only transgene, and seven mice (70%) had additional proto-oncogene inserts. We repeated the same experiment after removing MYC-related genes from the library to characterize additional leukemia-inducing gene combinations. Our second screen using the MYC-deleted proto-oncogene library confirmed MEIS1and the HOX family as cooperating oncogenes in leukemia pathogenesis. The model system we introduced in this study will be valuable in functionally screening novel combinations of genes for leukemogenic potential in vivo, and the system will help in the discovery of new targets for leukemia therapy.

  15. cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

    Directory of Open Access Journals (Sweden)

    F.H.R. Fagundes

    2010-03-01

    Full Text Available To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR, we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49, from the venom of Crotalus durissus collilineatus (Cdc PLA2. The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC. The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.

  16. Radioactive cDNA microarray (II): Gene expression profiling of antidepressant treatment by human cDNA microarray

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ji Hye; Kang, Rhee Hun; Ham, Byung Joo; Lee, Min Su; Shin, Kyung Ho; Choe, Jae Gol; Kim, Meyoung Kon [College of Medicine, Univ. of Korea, Seoul (Korea, Republic of)

    2003-07-01

    Major depressive disorder is a prevalent psychiatric disorder in primary care, associated with impaired patient functioning and well-being. Fluoxetine is a selective serotonin-reuptake inhibitors (SSRIs) and is a commonly prescribed antidepressant compound. Its action is primarily attributed to selective inhibition of the reuptake of serotonin (5-hydroxytryptamine) in the central nervous system. Objectives ; the aims of this study were two-fold: (1) to determine the usefulness for investigation of the transcription profiles in depression patients, and (2) to assess the differences in gene expression profiles between positive response group and negative response groups by fluoxetine treatment. This study included 53 patients with major depression (26 in positive response group with antidepressant treatment, 27 in negative response group with antidepressant treatment), and 53 healthy controls. To examine the difference of gene expression profile in depression patients, radioactive complementary DNA microarrays were used to evaluate changes in the expression of 1,152 genes in total. Using 33p-labeled probes, this method provided highly sensitive gene expression profiles including brain receptors, drug metabolism, and cellular signaling. Gene transcription profiles were classified into several categories in accordance with the antidepressant gene-regulation. The gene profiles were significantly up-(22 genes) and down-(16 genes) regulated in the positive response group when compared to the control group. Also, in the negative response group, 35 genes were up-regulated and 8 genes were down-regulated when compared to the control group. Consequently, we demonstrated that radioactive human cDNA microarray is highly likely to be an efficient technology for evaluating the gene regulation of antidepressants, such as selective serotonin-reuptake inhibitors (SSRIs), by using high-throughput biotechnology.

  17. Dissolved families

    DEFF Research Database (Denmark)

    Christoffersen, Mogens

    The situation in the family preceding a family separation is studied here, to identify risk factors for family dissolution. Information registers covering prospective statistics about health aspects, demographic variables, family violence, self-destructive behaviour, unemployment, and the spousal...

  18. Dissolved families

    DEFF Research Database (Denmark)

    Christoffersen, Mogens

    The situation in the family preceding a family separation is studied here, to identify risk factors for family dissolution. Information registers covering prospective statistics about health aspects, demographic variables, family violence, self-destructive behaviour, unemployment, and the spousal...

  19. Family Life

    Science.gov (United States)

    ... With Family and Friends > Family Life Request Permissions Family Life Approved by the Cancer.Net Editorial Board , ... your outlook on the future. Friends and adult family members The effects of cancer on your relationships ...

  20. Molecular cloning of the cDNA encoding follicle-stimulating hormone beta subunit of the Chinese soft-shell turtle Pelodiscus sinensis, and its gene expression.

    Science.gov (United States)

    Chien, Jung-Tsun; Shen, San-Tai; Lin, Yao-Sung; Yu, John Yuh-Lin

    2005-04-01

    Follicle-stimulating hormone (FSH) is a member of the pituitary glycoprotein hormone family. These hormones are composed of two dissimilar subunits, alpha and beta. Very little information is available regarding the nucleotide and amino acid sequence of FSHbeta in reptilian species. For better understanding of the phylogenetic diversity and evolution of FSH molecule, we have isolated and sequenced the complementary DNA (cDNA) encoding the Chinese soft-shell turtle (Pelodiscus sinensis, Family of Trionychidae) FSHbeta precursor molecule by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) methods. The cloned Chinese soft-shell turtle FSHbeta cDNA consists of 602-bp nucleotides, including 34-bp nucleotides of the 5'-untranslated region (UTR), 396-bp of the open reading frame, and 3'-UTR of 206-bp nucleotides. It encodes a 131-amino acid precursor molecule of FSHbeta subunit with a signal peptide of 20 amino acids followed by a mature protein of 111 amino acids. Twelve cysteine residues, forming six disulfide bonds within beta-subunit and two putative asparagine-linked glycosylation sites, are also conserved in the Chinese soft-shell turtle FSHbeta subunit. The deduced amino acid sequence of the Chinese soft-shell turtle FSHbeta shares identities of 97% with Reeves's turtle (Family of Bataguridae), 83-89% with birds, 61-70% with mammals, 63-66% with amphibians and 40-58% with fish. By contrast, when comparing the FSHbeta with the beta-subunits of the Chinese soft-shell turtle luteinizing hormone and thyroid stimulating hormone, the homologies are as low as 38 and 39%, respectively. A phylogenetic tree including reptilian species of FSHbeta subunits, is presented for the first time. Out of various tissues examined, FSHbeta mRNA was only expressed in the pituitary gland and can be up-regulated by gonadotropin-releasing hormone in pituitary tissue culture as estimated by fluorescence real-time PCR analysis.

  1. Laser cutting of triangular geometries in aluminum foam: Effect of cut size on thermal stress levels

    Science.gov (United States)

    Yilbas, B. S.; Akhtar, S. S.; Keles, O.

    2013-06-01

    Laser cutting of triangle shaped geometries in aluminum foam is carried out and the influence of triangle size on temperature and stress fields is examined. ABAQUS finite element code is used to simulate temperature and stress fields in the cut sections in line with the experimental conditions. Laser cut sections are examined incorporating optical and scanning electron microscopes, and energy dispersive spectroscopy. It is found that small size triangle cut results in relatively higher temperatures around the cut edges as compared to large size triangle cut; however, opposite is true for von Mises stress. This is attributed to the slow cooling rate in the case of the small size triangle cuts. Laser cut surfaces are free from large defects and locally scattered small dross attachment are found at the kerf exit.

  2. Selection of Near Optimal Laser Cutting Parameters in CO2 Laser Cutting by the Taguchi Method

    Directory of Open Access Journals (Sweden)

    Miloš MADIĆ

    2013-12-01

    Full Text Available Identification of laser cutting conditions that are insensitive to parameter variations and noise is of great importance. This paper demonstrates the application of Taguchi method for optimization of surface roughness in CO2 laser cutting of stainless steel. The laser cutting experiment was planned and conducted according to the Taguchi’s experimental design using the L27 orthogonal array. Four laser cutting parameters such as laser power, cutting speed, assist gas pressure, and focus position were considered in the experiment. Using the analysis of means and analysis of variance, the significant laser cutting parameters were identified, and subsequently the optimal combination of laser cutting parameter levels was determined. The results showed that the cutting speed is the most significant parameter affecting the surface roughness whereas the influence of the assist gas pressure can be neglected. It was observed, however, that interaction effects have predominant influence over the main effects on the surface roughness.

  3. Systematic Study of the Content of Phytochemicals in Fresh and Fresh-Cut Vegetables

    Directory of Open Access Journals (Sweden)

    María Isabel Alarcón-Flores

    2015-05-01

    Full Text Available Vegetables and fruits have beneficial properties for human health, because of the presence of phytochemicals, but their concentration can fluctuate throughout the year. A systematic study of the phytochemical content in tomato, eggplant, carrot, broccoli and grape (fresh and fresh-cut has been performed at different seasons, using liquid chromatography coupled to triple quadrupole mass spectrometry. It was observed that phenolic acids (the predominant group in carrot, eggplant and tomato were found at higher concentrations in fresh carrot than in fresh-cut carrot. However, in the case of eggplant, they were detected at a higher content in fresh-cut than in fresh samples. Regarding tomato, the differences in the content of phenolic acids between fresh and fresh-cut were lower than in other matrices, except in winter sampling, where this family was detected at the highest concentration in fresh tomato. In grape, the flavonols content (predominant group was higher in fresh grape than in fresh-cut during all samplings. The content of glucosinolates was lower in fresh-cut broccoli than in fresh samples in winter and spring sampling, although this trend changes in summer and autumn. In summary, phytochemical concentration did show significant differences during one-year monitoring, and the families of phytochemicals presented different behaviors depending on the matrix studied.

  4. Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence

    Institute of Scientific and Technical Information of China (English)

    WEIJi-cheng; YANGChuan-ping; WANGChao; JIANGJing

    2005-01-01

    Female inflorescence of Betula platyphylla was sampled at an interval of each two days to analyze the background of gene expression in floral phase. On the basis of SMART strategy, the driver cDNA was obtained from total RNA of the last sample and the tester cDNA was from that of the others by RT-PCR which were subsequently used to construct a subtracted cDNA library. The result of the ESTs (expression sequence tags) blastX showed that the genes in the subtracted cDNA library could be mainly clustered into 5 groups related to metabolism, transportation and signal transduction, cell cycle, stress response, and regulation. The relationship between gene expression and development was also discussed.

  5. Quantitative Transcript Analysis in Plants: Improved First-strand cDNA Synthesis

    Institute of Scientific and Technical Information of China (English)

    Nai-Zhong XIAO; Lei BA; Preben Bach HOLM; Xing-Zhi WANG; Steve BOWRA

    2005-01-01

    The quantity and quality of first-strand cDNA directly influence the accuracy of transcriptional analysis and quantification. Using a plant-derived α-tubulin as a model system, the effect of oligo sequence and DTT on the quality and quantity of first-strand cDNA synthesis was assessed via a combination of semi-quantitative PCR and real-time PCR. The results indicated that anchored oligo dT significantly improved the quantity and quality of α-tubulin cDNA compared to the conventional oligo dT. Similarly, omitting DTT from the first-strand cDNA synthesis also enhanced the levels of transcript. This is the first time that a comparative analysis has been undertaken for a plant system and it shows conclusively that small changes to current protocols can have very significant impact on transcript analysis.

  6. Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Dufva, I.H.; Dufva, Hans Martin

    2006-01-01

    oligonucleotides (pentadecamers) consistently, yielded at least 2 fold as much cDNA as did random hexamers using either-poly(A) RNA or an amplified version of messenger RNA (aRNA) as a template. The cDNA generated using pentadecamers did not differ in size distribution or the amount of incorporated label compared...... with cDNA generated with random hexamers. The increased efficiency of priming using random pentadecamers resulted in reverse transcription of > 80% of the template aRNA, while random hexamers induced reverse transcription of only 40% of the template aRNA. This suggests a better coverage...... that random pentadecamers can replace random hexamers in reverse transcription reactions on both poly(A) RNA and amplified RNA, resulting in higher cDNA yields and quality....

  7. Experimental research on the durability cutting tools for cutting-off steel profiles

    Directory of Open Access Journals (Sweden)

    Cristea Alexandru

    2017-01-01

    Full Text Available The production lines used for manufacturing U-shaped profiles are very complex and they must have high productivity. One of the most important stages of the fabrication process is the cutting-off. This paper presents the experimental research and analysis of the durability of the cutting tools used for cutting-off U-shaped metal steel profiles. The results of this work can be used to predict the durability of the cutting tools.

  8. Isolation of cosmid and cDNA clones in the region surrounding the BTK gene at Xq21.3-q22

    Energy Technology Data Exchange (ETDEWEB)

    Vorechovsky, I.; Zhou, J.N.; Hammarstroem, L. [Karolinska Institute, Huddinge (Sweden)] [and others

    1994-06-01

    A regional physical and transcription map involving yeast artificial chromosomes (YACs), cosmids, and cDNAs has been constructed for Xq21.3-q22 around the gene BTK (formerly atk or BPK) defective in X-linked agammaglobulinemia (XLA). With a positional cloning strategy employing direct cDNA selection, novel cDNAs were found to cluster in the region of approximately 100 kb flanking the XLA and {alpha}-galactosidase A loci. While these widely expressed transcripts are in the area known to contain CpG islands, a less evolutionarily conserved gene, located more than 130 kb distal of DXS178, maps to cosmid clones that could not be digested with rare-cutting restriction enzymes. The presence of transcribed sequences flanking the BTK allowed investigation of their involvement in complex XLA phenotypes. Southern blot analysis using cDNA clones isolated from this region permitted exclusion of a contiguous deletion syndrome as an underlying defect in three patients with XLA and associated growth hormone deficiency. A single XLA patient with torsion dystonia and cosegregating X-linked deafness has been found with a deletion in the 3{prime} part of BTK extending centromerically into the flanking expressed sequence DXS1274E. This suggests a possible involvement of the DXS1274E in this phenotype. The GenBank accession numbers for novel cDNA sequences are as follows: DXS1269E (L20773), DXS1271E (UO1923), DXS1273E (UO1925), and DXS1274E (UO1922). 51 refs., 4 figs., 1 tab.

  9. Conversion of cDNA differential display results (DDRT-PCR into quantitative transcription profiles

    Directory of Open Access Journals (Sweden)

    Koopmann Birger

    2005-04-01

    Full Text Available Abstract Background Gene expression studies on non-model organisms require open-end strategies for transcription profiling. Gel-based analysis of cDNA fragments allows to detect alterations in gene expression for genes which have neither been sequenced yet nor are available in cDNA libraries. Commonly used protocols for gel-based transcript profiling are cDNA differential display (DDRT-PCR and cDNA-AFLP. Both methods have been used merely as qualitative gene discovery tools so far. Results We developed procedures for the conversion of cDNA Differential Display data into quantitative transcription profiles. Amplified cDNA fragments are separated on a DNA sequencer and detector signals are converted into virtual gel images suitable for semi-automatic analysis. Data processing consists of four steps: (i cDNA bands in lanes corresponding to samples treated with the same primer combination are matched in order to identify fragments originating from the same transcript, (ii intensity of bands is determined by densitometry, (iii densitometric values are normalized, and (iv intensity ratio is calculated for each pair of corresponding bands. Transcription profiles are represented by sets of intensity ratios (control vs. treatment for cDNA fragments defined by primer combination and DNA mobility. We demonstrated the procedure by analyzing DDRT-PCR data on the effect of secondary metabolites of oilseed rape Brassica napus on the transcriptome of the pathogenic fungus Leptosphaeria maculans. Conclusion We developed a data processing procedure for the quantitative analysis of amplified cDNA fragments separated by electrophoresis. The system utilizes common software and provides an open-end alternative to DNA microarray analysis of the transcriptome. It is expected to work equally well with DDRT-PCR and cDNA-AFLP data and be useful particularly in reseach on organisms for which microarray analysis is not available or economical.

  10. Cloning of cDNA encoding steroid 11. beta. -hydroxylase (P450c11)

    Energy Technology Data Exchange (ETDEWEB)

    Chua, S.C.; Szabo, P.; Vitek, A.; Grzeschik, K.H.; John, M.; White, P.C.

    1987-10-01

    The authors have isolated bovine and human adrenal cDNA clones encoding the adrenal cytochrome P-450 specific for 11..beta..-hydroxylation (P450c11). A bovine adrenal cDNA library constructed in the bacteriophage lambda vector gt10 was probed with a previously isolated cDNA clone corresponding to part of the 3' untranslated region of the 4.2-kilobase (kb) mRNA encoding P450c11. Several clones with 3.2-kb cDNA inserts were isolated. Sequence analysis showed that they overlapped the original probe by 300 base pairs (bp). Combined cDNA and RNA sequence data demonstrated a continuous open reading frame of 1509 bases. P450c11 is predicted to contain 479 amino acid residues in the mature protein in addition to a 24-residue amino-terminal mitochondrial signal sequence. A bovine clone was used to isolate a homologous clone with a 3.5-kb insert from a human adrenal cDNA library. A region of 1100 bp was 81% homologous to 769 bp of the coding sequence of the bovine cDNA except for a 400-bp segment presumed to be an unprocessed intron. Hybridization of the human cDNA to DNA from a panel of human-rodent somatic cell hybrid lines and in situ hybridization to metaphase spreads of human chromosomes localized the gene to the middle of the long arm of chromosome 8. These data should be useful in developing reagents for heterozygote detection and prenatal diagnosis of 11..beta..-hydroxylase deficiency, the second most frequent cause of congenital adrenal hyperplasia.

  11. Cloning and Sequence Analysis of Interleukin 10 (IL-10) Full-length cDNA from Cyprinus carpio L.

    Institute of Scientific and Technical Information of China (English)

    Xiangru FENG; Yilong CHEN; Xiao ZHAO; Wendong WANG; Junhui ZHANG; Zhenguo YANG SUN; Shengmei JIA; Qiang LU

    2012-01-01

    Abstract [Objective] This study aimed to obtain IL-IO (interleukin 10) full-length cD- NA of common carp (Cyprinus carpio L.) and conduct the sequence analysis. []~lethod] The differentially expressed cDNA fragment was obtained by DD-RTPCR (differential display RT-PCR). The cDNA library of peripheral blood leukocytes which were separated from common carp and stimulated by mitogen was screened with a probe labeled with DIG (digoxigenin). The IL-IO full-length cDNA was cloned from 0.8x104 pfu of recombinant phages, and the sequence analysis and homology com- parison were carried out. [Result] Sequence analysis indicated that the IL-IO full- length cDNA of common carp was 1 117 bp long, containing a.55 bp 5'-UTR, a 522 bp 3"-UTR, and a 540 bp open reading frame(ORF) encoding 179 amino acids. In addition, there were three mRNA instability motifs (ATTTA) in the 3"-untranslated region. The deduced protein sequence shared typical sequence features of the IL-IO family. Homology comparison indicated that the obtained sequence shared 89.1% homology with the carp IL-IO gene from GenBank. [Conclusion] This study laid foun- dation for further study of the expression manner, functional characteristic and regu- lation mechanism of IL-IO in vivo and the interaction mechanism in the inflammatory reaction and immune response.

  12. Postharvest: Cut flowers and potted plants

    Science.gov (United States)

    In the past fifty years, the cut flower market has changed dramatically, from a local market with growers located on city outskirts, to a global one; flowers and cut foliage sourced from throughout the world are sold as bunches or combined into arrangements and bouquets in the major target markets. ...

  13. Performance Enhancement of Abrasive Waterjet Cutting

    NARCIS (Netherlands)

    Vu Ngoc Pi

    2008-01-01

    Abrasive Waterjet (AWJ) Machining is a recent non-traditional machining process. This technology is widely used in industry for cutting difficult-to-machine-materials, milling slots, polishing hard materials etc. AWJ machining has many advantages, e.g. it can cut net-shape parts, no heat is generate

  14. Experimental analysis of cut welding in aluminium

    DEFF Research Database (Denmark)

    Dorph, Pernille; De Chiffre, Leonardo; Bay, Niels

    1993-01-01

    Cut welding is a newly developed cold pressure welding process. In the present work, an experimental investigation was carried out analyzing the mechanisms involved in cut welding of a block to a strip. Experiments were carried out in technically pure aluminium. The investigation has involved...

  15. Designing for hot-blade cutting

    DEFF Research Database (Denmark)

    Brander, David; Bærentzen, Jakob Andreas; Clausen, Kenn

    2016-01-01

    -trivial constraints of blade-cutting in a bottom-up fashion, enabling an exploration of the unique architectural potential of this fabrication approach. The method is implemented as prototype design tools in MatLAB, C++, GhPython, and Python and demonstrated through cutting of expanded polystyrene foam design...

  16. Drilling subsurface wellbores with cutting structures

    Science.gov (United States)

    Mansure, Arthur James; Guimerans, Rosalvina Ramona

    2010-11-30

    A system for forming a wellbore includes a drill tubular. A drill bit is coupled to the drill tubular. One or more cutting structures are coupled to the drill tubular above the drill bit. The cutting structures remove at least a portion of formation that extends into the wellbore formed by the drill bit.

  17. Performance Enhancement of Abrasive Waterjet Cutting

    NARCIS (Netherlands)

    Vu Ngoc Pi

    2008-01-01

    Abrasive Waterjet (AWJ) Machining is a recent non-traditional machining process. This technology is widely used in industry for cutting difficult-to-machine-materials, milling slots, polishing hard materials etc. AWJ machining has many advantages, e.g. it can cut net-shape parts, no heat is

  18. Performance Enhancement of Abrasive Waterjet Cutting

    NARCIS (Netherlands)

    Vu Ngoc Pi

    2008-01-01

    Abrasive Waterjet (AWJ) Machining is a recent non-traditional machining process. This technology is widely used in industry for cutting difficult-to-machine-materials, milling slots, polishing hard materials etc. AWJ machining has many advantages, e.g. it can cut net-shape parts, no heat is generate

  19. Laser cutting - trends in the development,

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    2002-01-01

    Since the laser was invented in 1960, the industrial applications of this tool has grown and grown. And - since the beginning of the 1980'ies, the major industrial application of lasers in production has been laser cutting. In this paper a short review of the development of the laser cutting...

  20. Gluebond strength of laser cut wood

    Science.gov (United States)

    Charles W. McMillin; Henry A. Huber

    1985-01-01

    The degree of strength loss when gluing laser cut wood as compared to conventionally sawn wood and the amount of additional surface treatment needed to improve bond quality were assessed under normal furniture plant operating conditions. The strength of laser cut oak glued with polyvinyl acetate adhesive was reduced to 75 percent of sawn joints and gum was reduced 43...

  1. Laser cutting - trends in the development,

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove

    2002-01-01

    Since the laser was invented in 1960, the industrial applications of this tool has grown and grown. And - since the beginning of the 1980'ies, the major industrial application of lasers in production has been laser cutting. In this paper a short review of the development of the laser cutting...

  2. 33 CFR 117.143 - Bishop Cut.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Bishop Cut. 117.143 Section 117... OPERATION REGULATIONS Specific Requirements California § 117.143 Bishop Cut. The draw of the San Joaquin County highway bridge, mile 1.0 between King Island and Bishop Tract, shall open on signal if at least...

  3. Cutting Watermelon: Lessons in Instructional Coaching

    Science.gov (United States)

    Sandstead, Martha

    2016-01-01

    Literacy coordinator Martha Sandstead finds inspiration for her coaching work in a quote from civil rights organizer Lawrence Guyot: "Let's say you're riding past a picnic, and people are cuttin' watermelons. You don't immediately go and say, "stop the watermelon cutting" and let's talk. … You cut some watermelons, or you help…

  4. Cutting tool form compensation system and method

    Science.gov (United States)

    Barkman, W.E.; Babelay, E.F. Jr.; Klages, E.J.

    1993-10-19

    A compensation system for a computer-controlled machining apparatus having a controller and including a cutting tool and a workpiece holder which are movable relative to one another along a preprogrammed path during a machining operation utilizes a camera and a vision computer for gathering information at a preselected stage of a machining operation relating to the actual shape and size of the cutting edge of the cutting tool and for altering the preprogrammed path in accordance with detected variations between the actual size and shape of the cutting edge and an assumed size and shape of the cutting edge. The camera obtains an image of the cutting tool against a background so that the cutting tool and background possess contrasting light intensities, and the vision computer utilizes the contrasting light intensities of the image to locate points therein which correspond to points along the actual cutting edge. Following a series of computations involving the determining of a tool center from the points identified along the tool edge, the results of the computations are fed to the controller where the preprogrammed path is altered as aforedescribed. 9 figures.

  5. Cutting tool form compensaton system and method

    Science.gov (United States)

    Barkman, William E.; Babelay, Jr., Edwin F.; Klages, Edward J.

    1993-01-01

    A compensation system for a computer-controlled machining apparatus having a controller and including a cutting tool and a workpiece holder which are movable relative to one another along a preprogrammed path during a machining operation utilizes a camera and a vision computer for gathering information at a preselected stage of a machining operation relating to the actual shape and size of the cutting edge of the cutting tool and for altering the preprogrammed path in accordance with detected variations between the actual size and shape of the cutting edge and an assumed size and shape of the cutting edge. The camera obtains an image of the cutting tool against a background so that the cutting tool and background possess contrasting light intensities, and the vision computer utilizes the contrasting light intensities of the image to locate points therein which correspond to points along the actual cutting edge. Following a series of computations involving the determining of a tool center from the points identified along the tool edge, the results of the computations are fed to the controller where the preprogrammed path is altered as aforedescribed.

  6. Bun splitting: a practical cutting stock problem

    NARCIS (Netherlands)

    C.A. Glass (Catherine); J.M. van Oostrum (Jeroen)

    2010-01-01

    textabstractWe describe a new hierarchical 2D-guillotine Cutting Stock Problem. In contrast to the classic cutting stock problem, waste is not an issue. The problem relates to the removal of a defective part and assembly of the remaining parts into homogeneous size blocks. The context is the packing

  7. Cutting Watermelon: Lessons in Instructional Coaching

    Science.gov (United States)

    Sandstead, Martha

    2016-01-01

    Literacy coordinator Martha Sandstead finds inspiration for her coaching work in a quote from civil rights organizer Lawrence Guyot: "Let's say you're riding past a picnic, and people are cuttin' watermelons. You don't immediately go and say, "stop the watermelon cutting" and let's talk. … You cut some watermelons, or you help…

  8. Gene expression profile of esophageal cancer in North East India by cDNA microarray analysis

    Institute of Scientific and Technical Information of China (English)

    Indranil Chattopadhyay; Sujala Kapur; Joydeep Purkayastha; Rupkumar Phukan; Amal Kataki; Jagadish Mahanta; Sunita Saxena

    2007-01-01

    AIM: To identify alterations in genes and molecular functional pathways in esophageal cancer in a high incidence region of India where there is a widespread use of tobacco and betel quid with fermented areca nuts.METHODS: Total RNA was isolated from tumor and matched normal tissue of 16 patients with esophageal squamous cell carcinoma. Pooled tumor tissue RNA was labeled with Cy3-dUTP and pooled normal tissue RNA was labeled with Cy5-dUTP by direct labeling method.The labeled probes were hybridized with human 10K cDNA chip and expression profiles were analyzed by Genespring GX V 7.3 (Silicon Genetics).RESULTS: Nine hundred twenty three genes were differentially expressed. Of these, 611 genes were upregulated and 312 genes were downregulated. Using stringent criteria (P ≤ 0.05 and ≥ 1.5 fold change),127 differentially expressed genes (87 upregulated and 40 downregulated) were identified in tumor tissue. On the basis of Gene Ontology, four different molecular functional pathways (MAPK pathway,G-protein coupled receptor family, ion transport activity,and serine or threonine kinase activity) were most significantly upregulated and six different molecular functional pathways (structural constituent of ribosome,endopeptidase inhibitor activity, structural constituent of cytoskeleton, antioxidant activity, acyl group transferase activity, eukaryotic translation elongation factor activity)were most significantly downregulated.CONCLUSION: Several genes that showed alterations in our study have also been reported from a high incidence area of esophageal cancer in China. This indicates that molecular profiles of esophageal cancer in these two different geographic locations are highly consistent.

  9. A peroxiredoxin cDNA from Taiwanofungus camphorata: role of Cys31 in dimerization.

    Science.gov (United States)

    Huang, Chih-Yu; Chen, Yu-Ting; Wen, Lisa; Sheu, Dey-Chyi; Lin, Chi-Tsai

    2014-01-01

    Peroxiredoxins (Prxs) play important roles in antioxidant defense and redox signaling pathways. A Prx isozyme cDNA (TcPrx2, 745 bp, EF552425) was cloned from Taiwanofungus camphorata and its recombinant protein was overexpressed. The purified protein was shown to exist predominantly as a dimer by sodium dodecyl sulfate-polyacrylamide gel electrolysis in the absence of a reducing agent. The protein in its dimeric form showed no detectable Prx activity. However, the protein showed increased Prx activity with increasing dithiothreitol concentration which correlates with dissociation of the dimer into monomer. The TcPrx2 contains two Cys residues. The Cys(60) located in the conserved active site is the putative active peroxidatic Cys. The role of Cys(31) was investigated by site-directed mutagenesis. The C31S mutant (C(31) → S(31)) exists predominantly as a monomer with noticeable Prx activity. The Prx activity of the mutant was higher than that of the corresponding wild-type protein by nearly twofold at 12 μg/mL. The substrate preference of the mutant was H2O2 > cumene peroxide > t-butyl peroxide. The Michaelis constant (K M) value for H2O2 of the mutant was 0.11 mM. The mutant enzyme was active under a broad pH range from 6 to 10. The results suggest a role of Cys(31) in dimerization of the TcPrx2, a role which, at least in part, may be involved in determining the activity of Prx. The C(31) residue does not function as a resolving Cys and therefore the TcPrx2 must follow the reaction mechanism of 1-Cys Prx. This TcPrx2 represents a new isoform of Prx family.

  10. ASSOCIATION OF DIFFERENTIALLY EXPRESSED cDNA FRAGMENT OF FGG WITH HEPATOCELLULAR CARCINOMA

    Institute of Scientific and Technical Information of China (English)

    范秉琳; 朱武凌; 邹国林; 段芳龄

    2002-01-01

    Objective: To identify a cDNA clone from the subtracted library of human hepatocellular carcinoma (HCC). Methods: Suppression subtractive hybridization was used to isolated a panel of genes that are differentially expressed in hepatocellular carcinoma as compared with cirrhotic liver. T/A cloning method was used to construct a subtracted cDNA library. DNA sequencing analysis and Northern blot analysis were also utilized. Results: The cloned cDNA is 787 nucleotides in length and contains an open reading frame of 230 amino acids, which is a cDNA fragment of reported human fibrinogen, gamma polypeptide (FGG). Northern analysis revealed that this gene was overexpressed in two hepatocellular carcinoma cell lines, SMMC-7721 and HepG2. Conclusion: Sequence identity proved the cDNA clone fragment of as FGG gene. Differential expression of the cDNA fragment in HCC suggested that FGG is related to HCC, indicating a new clue for developing a novel diagnostic and prognostic marker.

  11. Purification of Single-Stranded cDNA Based on RNA Degradation Treatment and Adsorption Chromatography.

    Science.gov (United States)

    Trujillo-Esquivel, Elías; Franco, Bernardo; Flores-Martínez, Alberto; Ponce-Noyola, Patricia; Mora-Montes, Héctor M

    2016-08-02

    Analysis of gene expression is a common research tool to study networks controlling gene expression, the role of genes with unknown function, and environmentally induced responses of organisms. Most of the analytical tools used to analyze gene expression rely on accurate cDNA synthesis and quantification to obtain reproducible and quantifiable results. Thus far, most commercial kits for isolation and purification of cDNA target double-stranded molecules, which do not accurately represent the abundance of transcripts. In the present report, we provide a simple and fast method to purify single-stranded cDNA, exhibiting high purity and yield. This method is based on the treatment with RNase H and RNase A after cDNA synthesis, followed by separation in silica spin-columns and ethanol precipitation. In addition, our method avoids the use of DNase I to eliminate genomic DNA from RNA preparations, which improves cDNA yield. As a case report, our method proved to be useful in the purification of single-stranded cDNA from the pathogenic fungus Sporothrix schenckii.

  12. [Rapid construction of full-length MnSOD cDNA of chickens by one-step 3'RACE].

    Science.gov (United States)

    Bu, You-Quan; Luo, Xu-Gang; Liu, Bin; Li, Su-Fen

    2004-07-01

    RACE (rapid amplification of cDNA ends) is a popular technique to rapidly obtain the full-length cDNA. After obtaining the 3' cDNA and 5' cDNA fragments with a overlapped region by 3' RACE and 5' RACE, the full-length cDNA could be generated by end-to-end PCR or subcloning. In this study, 3' RACE combined with touch-down PCR was successfully used for the rapid construction of full-length MnSOD cDNA of chickens. Compared with the conventional end-to-end PCR or subcloning, this method, called one-step 3' RACE, is fast, economical and highly specific. It especially fits the rapid construction of full-length cDNA by RACE method.

  13. Thermophysical problems of laser cutting of metals

    Directory of Open Access Journals (Sweden)

    Orishich Anatoliy

    2017-01-01

    Full Text Available Variety and complex interaction of physical processes during laser cutting is a critical characteristic of the laser cutting of metals. Small spatial and temporal scales complicate significantly the experimental investigations of the multi-phase fluid flow in the conditions of laser cutting of metals. In these conditions, the surface formed during the cutting is an indicator determining the melt flow character. The quantitative parameter reflecting the peculiarities of the multi-phase fluid flow, is normally the roughness of the forming surface, and the minimal roughness is the criterion of the qualitative flow [1 – 2]. The purpose of this work is to perform the experimental comparative investigation of the thermophysical pattern of the multi-phase melt flow in the conditions of the laser cutting of metals with the laser wavelength of 10.6 μm and 1.07 μm.

  14. Cutting as a continuous business process

    Directory of Open Access Journals (Sweden)

    Miro Gradišar

    2009-11-01

    Full Text Available A review of state-of-the-art methods for cutting stock problem optimisation shows that the current methods lead to near-optimum results for the instantaneous optimisation of trim loss. Further optimisation of this activity would not bring a considerable improvement. Therefore, the paper treats cutting stock as a continuous business process and not as an isolated activity. An exact method for a general one-dimensional cutting stock problem is presented and tested. The method is mainly suitable for smaller orders. It is then applied to continuous cutting and used to develop a method for assessing cutting costs in consecutive time periods. The modified method finds a good solution over the whole time-span, rather than just local optima.

  15. Corn stalk orientation effect on mechanical cutting

    Energy Technology Data Exchange (ETDEWEB)

    Igathinathane, C. [Mississippi State University (MSU); Womac, A.R. [University of Tennessee; Sokhansanj, Shahabaddine [ORNL

    2010-07-01

    Research efforts that increase the efficiency of size reduction of biomass can lead to a significant energy saving. This paper deals with the determination of the effect of sample orientation with respect to cutting element and quantify the possible cutting energy reduction, utilising dry corn stalks as the test material (15%e20% wet basis). To evaluate the mechanical cutting characteristics of corn stalks, a Warnere Bratzler device was modified by replacing its blunt edged cutting element with one having a 30_ single bevel sharp knife edge. Cutting force-deformation characteristics obtained with a universal testing machine were analysed to evaluate the orientation effects at perpendicular (90o), inclined (45o), and parallel (0o) orientations on internodes and nodes for cutting force, energy, ultimate stress, and specific energy of corn stalks. The corn stalks cutting force-displacement characteristics were found to differ with orientation, and internode and node material difference. Overall, the peak failure force, and the total cutting energy of internodes and nodes varied significantly (P < 0.05) with stalk cross-sectional area. The specific energy values (total energy per unit cut area) of dry corn stalk internodes ranged from 11.3 to 23.5 kN m_1, and nodes from 8.6 to 14.0 kN m_1. The parallel orientation (along grain) compared to perpendicular (across grain) produced a significant reduction of the cutting stress and the specific energy to one tenth or better for internodes, and to about one-fifth for nodes.

  16. Laser cutting of Kevlar laminates and thermal stress formed at cutting sections

    Science.gov (United States)

    Yilbas, B. S.; Akhtar, S. S.

    2012-02-01

    Laser cutting of Kevlar laminates is carried out and thermal stress field developed in the cutting region is predicted using the finite element code. Temperature predictions are validated through the thermocouple data. The morphological changes in the cutting section are examined by incorporating optical and scanning electron microscopes. It is found that temperature predictions agree well with the thermocouple data. High values of von Mises stress are observed at the cutting edges and at the mid-thickness of the Kevlar laminate due to thermal compression formed in this region. The laser cut edges are free from whiskers; however, striation formation and some small sideways burning is observed at the kerf edges.

  17. USA aborts international family planning.

    Science.gov (United States)

    Potts, M

    1996-03-02

    The US Agency for International Development (USAID) has been a leader in international family planning for almost 30 years, accounting for 46% of all funds in international family planning provided by OECD countries during 1991. Moreover, relative to other donor countries, the US supplies worldwide a disproportionate amount of contraceptives. While international family planning activities received $546 million in 1995, the budget was slashed in 1996 to $72 million. This unprecedented cut will have a profound effect upon the reproductive health and family planning choices of tens of millions of people in developing countries. Millions of additional unintended pregnancies and maternal and child deaths may result. 1996 began with the White House and Congress in political gridlock, with negotiations on foreign aid stalled on the issue of abortion. The Republican-led House of Representatives wanted to bar support of any nongovernmental organization (NGO) which also provided information on abortion, while Democratic President Bill Clinton affirmed that he would veto such legislation. At the end of January, the House passed the Balanced Budget and Down Payment Act (HR 2880) containing clauses which cut the aid budget by 35% and barring new money in the area of family planning until July 1. Spending was limited to the allocation of 6.5% of the total budget each month. Some social marketing programmers who distribute condoms and oral contraceptives are already feeling the pinch, and some programs will simply run out of contraceptives. This cut in funding also bodes ill for achieving the goals of the 1994 International Conference on Population and Development. There is, however, hope that the cuts will be reversed for the next fiscal year. The author notes survey findings which indicate that US citizens support higher budgets for family planning.

  18. Estimating cutting front temperature difference in disk and CO2 laser beam fusion cutting

    Science.gov (United States)

    Scintilla, L. D.; Tricarico, L.

    2012-07-01

    A three-dimensional, semi-stationary, simplified thermal numerical model was developed. The average cutting front temperature difference in disk and CO2 laser beam fusion cutting of 90MnCrV8 was estimated by computing the conductive power loss. Basing on heat affected zone extension experimentally measured and using an inverse methodology approach, the unknown thermal load on the cutting front during laser cutting was calculated. The accuracy of the numerical power loss estimation was evaluated comparing the results from simulation with the ones from analytical models. A good agreement was found for all the test cases considered in this study. The conduction losses estimation was used for justifying the lower quality of disk laser cuts due to the lower average cut front temperature. This results in the increase of viscosity of molten material and in the subsequent more difficult ejection of the melted material from the cut kerf.

  19. Results of investigations into coal cutting by asymmetric disks

    Energy Technology Data Exchange (ETDEWEB)

    Krauze, K.

    1985-02-01

    Effects are analyzed of design and specifications of asymmetric disk cutters on coal cutting by a shearer loader with disk cutters on helical cutting drums. Effects of disk diameter, wedge angle, cutting depth and chip thickness on cutting resistance were analyzed under operational conditions (a coal seam was cut by asymmetric disk cutters). On the basis of analysis of coal resistance to cutting by asymmetric cutting disks, regression equations were derived which describe coal cutting. Effects of disk parameters and cutting conditions on cutting resistance were determined. Analyses show that replacing radial cutting tools with asymmetric disk cutters would cause an increase in energy consumption of cutting and increase coal resistance to cutting. 1 reference.

  20. cDNA cloning and bacterial expression of a PL-14 alginate lyase from a herbivorous marine snail Littorina brevicula.

    Science.gov (United States)

    Rahman, Mohammad Matiur; Wang, Ling; Inoue, Akira; Ojima, Takao

    2012-10-01

    Herbivorous marine snails like Littorina species are known to possess alginate lyases in their digestive tracts. The Littorina enzymes have been identified as endolytic polymannuronate (poly(M)) lyases (EC 4.2.2.3); however, it is still unclear which polysaccharide-lyase family (PL) the Littorina enzymes belong to, since no complete primary structure of Littorina enzymes has been determined. Thus, in the present study, we analyzed the primary structure of LbAly28, a 28kDa alginate lyase isozyme of Littorina brevicula, by the cDNA method. LbAly28 cDNAs were amplified by PCR followed by 5'- and 3'-RACE PCRs from the L. brevicula hepatopancreas cDNA. A cDNA covering entire coding region of LbAly28 consisted of 1129bp and encoded an amino-acid sequence of 291 residues. The deduced amino-acid sequence comprised an initiation methionine, a putative signal peptide of 14 residues, a propeptide-like region of 16 residues, and a mature LbAly28 domain of 260 residues. The mature LbAly28 domain showed 43-53% amino-acid identities with other molluscan PL-14 enzymes. The catalytically important residues in PL-14 enzymes, which were identified in the Chlorella virus glucuronate-specific lyase vAL-1 and Aplysia poly(M) lyase AkAly30, were also conserved in LbAly28. Site-directed mutagenesis regarding these residues, that is, replacements of Lys94, Lys97, Thr121, Arg 123, Tyr135, and Tyr137 to Ala, decreased the activity of recombinant LbAly28 to various degrees. From these results we concluded that LbAly28 is a member of PL-14 alginate lyases. Besides the effects of above mutations, we noticed that the replacement of T121 by Ala changed the substrate preference of LbAly28. Namely, the activities toward sodium alginate and poly(MG)-block substrate increased and became comparable with the activity toward poly(M)-block substrate. This suggests that the region including T121 of LbAly28 closely relates to the recognition of poly(MG) region of alginate.

  1. Rejuvenated sycamore cuttings for energy plantations

    Energy Technology Data Exchange (ETDEWEB)

    Land, S.B. Jr.; Elam, W.W. [Mississippi State Univ., MS (United States). Dept. of Forestry; Khan, Mohammad [Pakistan Forest Inst., Peshawar (Pakistan). Forestry Research Div.

    1995-12-31

    Rejuvination of mature, progeny tested American sycamore trees is needed to increase rooting success of cuttings for energy plantations. Studies conducted in Mississippi (U.S.A.) indicated that hedging of stock plants and spraying the stock plants with benzyl adenine may rejuvinate the cuttings. Percentages of cuttings with early bud break in one study and with sprouts and roots after nine months in another study were increased. The rooted cuttings grew rapidly like juvenile material and showed no plagiotrophic effects during the first year after transplanting. At the end of that year the dry weight was 2.13 Mg ha{sup -1} (1.2 m x 3.6 m spacing) and was distributed in stems (31%), branches (14%), leaves (24%) and roots (31%). An IBA rooting powder decreased rooting success. Planting cuttings directly in the nursery, rather than first in the greenhouse, increased success when frost was absent. Use of cuttings of greater than 14 m diameter with a node within 2 cm of the base increased success. Clonal differences in frost susceptibility of sprouting cuttings were observed. (author)

  2. New Tool Coatings for Light Metal Cutting

    Institute of Scientific and Technical Information of China (English)

    DaveDoerwald; ThomasKrug; RoelTietema; Wei-MingSim; QuanshunLuo; PapkenHovsepian

    2004-01-01

    New carbon based coatings have been developed for cutting of light metals such as aluminum. These coatings can be used either directly on the tool or as topcoating combined with another hard base coating. Cutting tests show that the coating gives good performance for materials that tend to stick to the cutting tools, such as aluminium alloys, because of the substantial reduction of appearance of the so-called built-up edge (BUE). This results in a longer life time of the tool and smoother surface finish of the cut made in the work-piece material. Especially in dry machining and deep hole drilling this coating performed very well. An overview of the properties and initial cutting results of this coating will be presented. This paper will also present results from recent industrial field tests carried out by a leading European aerospace manufacturer. Cutting forces, BUE formation and surface roughness data will be presented to explain the cutting process during dry machining.

  3. New Tool Coatings for Light Metal Cutting

    Institute of Scientific and Technical Information of China (English)

    Dave Doerwald; Thomas Krug; Roel Tietema; Wei-Ming Sim; Quanshun Luo; Papken Hovsepian

    2004-01-01

    New carbon based coatings have been developed for cutting of light metals such as aluminum. These coatings can be used either directly on the tool or as topcoating combined with another hard base coating. Cutting tests show that the coating gives good performance for materials that tend to stick to the cutting tools, such as aluminium alloys, because of the substantial reduction of appearance of the so-called built-up edge (BUE). This results in a longer life time of the tool and smoother surface finish of the cut made in the work-piece material. Especially in dry machining and deep hole drilling this coating performed very well. An overview of the properties and initial cutting results of this coating will be presented. This paper will also present results from recent industrial field tests carried out by a leading European aerospace manufacturer.Cutting forces, BUE formation and surface roughness data will be presented to explain the cutting process during dry machining.

  4. Laser cutting of sheets for Tailored Blanks

    DEFF Research Database (Denmark)

    Bagger, Claus; Olsen, Flemming Ove

    1999-01-01

    Over the past few years there has been an enormous increase in the use of tailored blanks, especially in the automotive industry. Often the sheets for tailored blanks are shear cut, but results have been reported that the allowable sheet gap distance should not exceed 0.1 mm in order to obtain...... sound welds. Laser cutting the sheets may therefore be an alternative to shear cutting, if the cut kerf squareness can be kept below 0.05 mm.In a number of systematic laboratory experiments the effects of the major process parameters in laser cutting have been investigated. Each cut was quantified...... by the squareness, the surface roughness and the burr height. Mild steel as well as high strength steel with and with out galvanisation with thickness' of 0.7(5) and 1.25 were used.In the tests the difference in cut quality between a 5" and a 7.5" focusing lens were tested and the effect of using pulsed mode laser...

  5. Reachability cuts for the vehicle routing problem with time windows

    DEFF Research Database (Denmark)

    Lysgaard, Jens

    2004-01-01

    This paper introduces a class of cuts, called reachability cuts, for the Vehicle Routing Problem with Time Windows (VRPTW). Reachability cuts are closely related to cuts derived from precedence constraints in the Asymmetric Traveling Salesman Problem with Time Windows and to k-path cuts...

  6. Reachability cuts for the vehicle routing problem with time windows

    DEFF Research Database (Denmark)

    Lysgaard, Jens

    2004-01-01

    This paper introduces a class of cuts, called reachability cuts, for the Vehicle Routing Problem with Time Windows (VRPTW). Reachability cuts are closely related to cuts derived from precedence constraints in the Asymmetric Traveling Salesman Problem with Time Windows and to k-path cuts...

  7. Cloning and screening of cDNA of Psilgramma menephorn allergen

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To construct a cDNA expression library of Psilgramma menephorn to screen its major allergen so as to provide the basis for producing recombinant allergen vaccine of Psilgramma menephorn. Methods Total RNA was extracted from the whole body of Psilgramma menephorn with Trizol and mRNA was purified with Oligo (dT) Spin-Column. And dscDNA was synthesized through reverse transcription. After blunting, the cDNA fragments were ligated with EcoRⅠ adapters. Then the cDNAs were digested by XhoⅠ, and the fragments less than 400 bp were removed by using GHROMA SPIN-400 column. The remaining fragments longer than 400 bp were ligated with Uni-ZAP XR vector. The recombinants were packaged in vitro and a small portion of the packaged phage was used to infect E.coli XL1-Blue MRF′ for titration. The recombinants were examined by color selection. The size of cDNA inserts and the diversity of library were analyzed by PCR. The library was screened using SPT positive sera from patients with Psilgramma menephorn allergy repeatedly. Results The cDNA expression library consisting of a 5×105 recombinant bacteriophages was constructed with the recombinant ratio of 67%. The average length of recombinant exogenous inserts was about 1.49 kb. Five positive cDNA clones were obtained. Conclusion The constructed cDNA expression library shows appropriate contents and size of cDNA fragments and the related genes of Psilgramma menephorn major allergens were harbored successfully, which lays the foundation for the positive clone identification and further analysis.

  8. The effect of column purification on cDNA indirect labelling for microarrays

    Directory of Open Access Journals (Sweden)

    Kiss John Z

    2007-06-01

    Full Text Available Abstract Background The success of the microarray reproducibility is dependent upon the performance of standardized procedures. Since the introduction of microarray technology for the analysis of global gene expression, reproducibility of results among different laboratories has been a major problem. Two of the main contributors to this variability are the use of different microarray platforms and different laboratory practices. In this paper, we address the latter question in terms of how variation in one of the steps of a labelling procedure affects the cDNA product prior to microarray hybridization. Results We used a standard procedure to label cDNA for microarray hybridization and employed different types of column chromatography for cDNA purification. After purifying labelled cDNA, we used the Agilent 2100 Bioanalyzer and agarose gel electrophoresis to assess the quality of the labelled cDNA before its hybridization onto a microarray platform. There were major differences in the cDNA profile (i.e. cDNA fragment lengths and abundance as a result of using four different columns for purification. In addition, different columns have different efficiencies to remove rRNA contamination. This study indicates that the appropriate column to use in this type of protocol has to be experimentally determined. Finally, we present new evidence establishing the importance of testing the method of purification used during an indirect labelling procedure. Our results confirm the importance of assessing the quality of the sample in the labelling procedure prior to hybridization onto a microarray platform. Conclusion Standardization of column purification systems to be used in labelling procedures will improve the reproducibility of microarray results among different laboratories. In addition, implementation of a quality control check point of the labelled samples prior to microarray hybridization will prevent hybridizing a poor quality sample to expensive

  9. cDNA2Genome: A tool for mapping and annotating cDNAs

    Directory of Open Access Journals (Sweden)

    Suhai Sandor

    2003-09-01

    Full Text Available Abstract Background In the last years several high-throughput cDNA sequencing projects have been funded worldwide with the aim of identifying and characterizing the structure of complete novel human transcripts. However some of these cDNAs are error prone due to frameshifts and stop codon errors caused by low sequence quality, or to cloning of truncated inserts, among other reasons. Therefore, accurate CDS prediction from these sequences first require the identification of potentially problematic cDNAs in order to speed up the posterior annotation process. Results cDNA2Genome is an application for the automatic high-throughput mapping and characterization of cDNAs. It utilizes current annotation data and the most up to date databases, especially in the case of ESTs and mRNAs in conjunction with a vast number of approaches to gene prediction in order to perform a comprehensive assessment of the cDNA exon-intron structure. The final result of cDNA2Genome is an XML file containing all relevant information obtained in the process. This XML output can easily be used for further analysis such us program pipelines, or the integration of results into databases. The web interface to cDNA2Genome also presents this data in HTML, where the annotation is additionally shown in a graphical form. cDNA2Genome has been implemented under the W3H task framework which allows the combination of bioinformatics tools in tailor-made analysis task flows as well as the sequential or parallel computation of many sequences for large-scale analysis. Conclusions cDNA2Genome represents a new versatile and easily extensible approach to the automated mapping and annotation of human cDNAs. The underlying approach allows sequential or parallel computation of sequences for high-throughput analysis of cDNAs.

  10. cDNA sequencing improves the detection of P53 missense mutations in colorectal cancer

    Directory of Open Access Journals (Sweden)

    Jesionek-Kupnicka Dorota

    2009-08-01

    Full Text Available Abstract Background Recently published data showed discrepancies beteween P53 cDNA and DNA sequencing in glioblastomas. We hypothesised that similar discrepancies may be observed in other human cancers. Methods To this end, we analyzed 23 colorectal cancers for P53 mutations and gene expression using both DNA and cDNA sequencing, real-time PCR and immunohistochemistry. Results We found P53 gene mutations in 16 cases (15 missense and 1 nonsense. Two of the 15 cases with missense mutations showed alterations based only on cDNA, and not DNA sequencing. Moreover, in 6 of the 15 cases with a cDNA mutation those mutations were difficult to detect in the DNA sequencing, so the results of DNA analysis alone could be misinterpreted if the cDNA sequencing results had not also been available. In all those 15 cases, we observed a higher ratio of the mutated to the wild type template by cDNA analysis, but not by the DNA analysis. Interestingly, a similar overexpression of P53 mRNA was present in samples with and without P53 mutations. Conclusion In terms of colorectal cancer, those discrepancies might be explained under three conditions: 1, overexpression of mutated P53 mRNA in cancer cells as compared with normal cells; 2, a higher content of cells without P53 mutation (normal cells and cells showing K-RAS and/or APC but not P53 mutation in samples presenting P53 mutation; 3, heterozygous or hemizygous mutations of P53 gene. Additionally, for heterozygous mutations unknown mechanism(s causing selective overproduction of mutated allele should also be considered. Our data offer new clues for studying discrepancy in P53 cDNA and DNA sequencing analysis.

  11. Cut elimination in multifocused linear logic

    DEFF Research Database (Denmark)

    Guenot, Nicolas; Brock-Nannestad, Taus

    2015-01-01

    We study cut elimination for a multifocused variant of full linear logic in the sequent calculus. The multifocused normal form of proofs yields problems that do not appear in a standard focused system, related to the constraints in grouping rule instances in focusing phases. We show that cut...... elimination can be performed in a sensible way even though the proof requires some specific lemmas to deal with multifocusing phases, and discuss the difficulties arising with cut elimination when considering normal forms of proofs in linear logic....

  12. Cut elimination in multifocused linear logic

    DEFF Research Database (Denmark)

    Guenot, Nicolas; Brock-Nannestad, Taus

    2015-01-01

    We study cut elimination for a multifocused variant of full linear logic in the sequent calculus. The multifocused normal form of proofs yields problems that do not appear in a standard focused system, related to the constraints in grouping rule instances in focusing phases. We show that cut elim...... elimination can be performed in a sensible way even though the proof requires some specific lemmas to deal with multifocusing phases, and discuss the difficulties arising with cut elimination when considering normal forms of proofs in linear logic....

  13. Reconfigurable manufacturing execution system for pipe cutting

    Science.gov (United States)

    Yin, Y. H.; Xie, J. Y.

    2011-08-01

    This article presents a reconfigurable manufacturing execution system (RMES) filling the gap between enterprise resource planning and resource layer for pipe-cutting production with mass customisation and rapid adaptation to dynamic market, which consists of planning and scheduling layer and executive control layer. Starting from customer's task and process requirements, the cutting trajectories are planned under generalised mathematical model able to reconfigure in accordance with various intersecting types' joint, and all tasks are scheduled by nesting algorithm to maximise the utilisation rate of rough material. This RMES for pipe cutting has been effectively implemented in more than 100 companies.

  14. The venom gland transcriptome of Latrodectus tredecimguttatus revealed by deep sequencing and cDNA library analysis.

    Directory of Open Access Journals (Sweden)

    Quanze He

    Full Text Available Latrodectus tredecimguttatus, commonly known as black widow spider, is well known for its dangerous bite. Although its venom has been characterized extensively, some fundamental questions about its molecular composition remain unanswered. The limited transcriptome and genome data available prevent further understanding of spider venom at the molecular level. In the present study, we combined next-generation sequencing and conventional DNA sequencing to construct a venom gland transcriptome of the spider L. tredecimguttatus, which resulted in the identification of 9,666 and 480 high-confidence proteins among 34,334 de novo sequences and 1,024 cDNA sequences, respectively, by assembly, translation, filtering, quantification and annotation. Extensive functional analyses of these proteins indicated that mRNAs involved in RNA transport and spliceosome, protein translation, processing and transport were highly enriched in the venom gland, which is consistent with the specific function of venom glands, namely the production of toxins. Furthermore, we identified 146 toxin-like proteins forming 12 families, including 6 new families in this spider in which α-LTX-Lt1a family2 is firstly identified as a subfamily of α-LTX-Lt1a family. The toxins were classified according to their bioactivities into five categories that functioned in a coordinate way. Few ion channels were expressed in venom gland cells, suggesting a possible mechanism of protection from the attack of their own toxins. The present study provides a gland transcriptome profile and extends our understanding of the toxinome of spiders and coordination mechanism for toxin production in protein expression quantity.

  15. Extreme Family Poverty Rises in the Midst of a Strong Economy. Family Review.

    Science.gov (United States)

    Lindjord, Denise

    2000-01-01

    Discusses the sudden increase of families with incomes below 50 percent of the poverty line, noting particularly the impact on children. Notes a direct link between increased family poverty and the decreasing protective role of cash welfare and food stamp benefits. Describes federal proposals to increase minimum wage and cut taxes; makes…

  16. Family Therapy

    Science.gov (United States)

    ... may be credentialed by the American Association for Marriage and Family Therapy (AAMFT). Family therapy is often short term. ... challenging situations in a more effective way. References Marriage and family therapists: The friendly mental health professionals. American Association ...

  17. Familial hypertriglyceridemia

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/000397.htm Familial hypertriglyceridemia To use the sharing features on this page, please enable JavaScript. Familial hypertriglyceridemia is a common disorder passed down through families. ...

  18. Family Meals

    Science.gov (United States)

    ... Teaching Kids to Be Smart About Social Media Family Meals KidsHealth > For Parents > Family Meals Print A ... even more important as kids get older. Making Family Meals Happen It can be a big challenge ...

  19. Family Arguments

    Science.gov (United States)

    ... Spread the Word Shop AAP Find a Pediatrician Family Life Medical Home Family Dynamics Adoption & Foster Care ... Life Listen Español Text Size Email Print Share Family Arguments Page Content Article Body We seem to ...

  20. Family History

    Science.gov (United States)

    Your family history includes health information about you and your close relatives. Families have many factors in common, including their genes, ... as heart disease, stroke, and cancer. Having a family member with a disease raises your risk, but ...

  1. Improved performance of linear coal cutting compared with rotary cutting. Report of investigations/1995

    Energy Technology Data Exchange (ETDEWEB)

    Roepke, W.W.; Hanson, B.D.; Olson, R.C.; Wingquist, C.F.; Myren, T.A.

    1995-07-01

    The linear cutting system, developed by the U.S. Bureau of Mines, uses geometric principles developed by Cardan to produce a nearly constant cut depth. The new system has been extensively tested in a synthetic material under laboratory conditions to verify mechanical capability and to identify operational characteristics. Comparisons between 15-rpm linear cutting and 50-rpm rotary cutting systems show significant improvement in respirable dust entrainment, product size distribution, and energy usage. Respirable dust is reduced by as much as 90 pct. Recovered product showed a 67-pct reduction in -0.32-cm (-1/8-in) material and a 200-pct increase in +5.08 cm (+2 in) material. Average power was reduced by 66 pct for the linear cutting. Because the bit cutting paths differ between linear and rotary cutting, it was necessary to compare the two at the same cut depths and bit types. These comparisons show that low revolution per minute rotary cutting entrains about the same amount of respirable dust as the linear cutting system, but the average shaft torque may be 55 to 130 pct greater for the rotary system.

  2. An FMS Dynamic Production Scheduling Algorithm Considering Cutting Tool Failure and Cutting Tool Life

    Science.gov (United States)

    Setiawan, A.; Wangsaputra, R.; Martawirya, Y. Y.; Halim, A. H.

    2016-02-01

    This paper deals with Flexible Manufacturing System (FMS) production rescheduling due to unavailability of cutting tools caused either of cutting tool failure or life time limit. The FMS consists of parallel identical machines integrated with an automatic material handling system and it runs fully automatically. Each machine has a same cutting tool configuration that consists of different geometrical cutting tool types on each tool magazine. The job usually takes two stages. Each stage has sequential operations allocated to machines considering the cutting tool life. In the real situation, the cutting tool can fail before the cutting tool life is reached. The objective in this paper is to develop a dynamic scheduling algorithm when a cutting tool is broken during unmanned and a rescheduling needed. The algorithm consists of four steps. The first step is generating initial schedule, the second step is determination the cutting tool failure time, the third step is determination of system status at cutting tool failure time and the fourth step is the rescheduling for unfinished jobs. The approaches to solve the problem are complete-reactive scheduling and robust-proactive scheduling. The new schedules result differences starting time and completion time of each operations from the initial schedule.

  3. Definition of fine cutting features for laser fusion cutting of stainless steel

    Science.gov (United States)

    Seebach, J.; Norman, S.; Harrison, P.

    2015-07-01

    Laser fusion cutting of stainless steel is often considered in a material range from 0,3mm up to 4mm and laser powers up to 2kW. For a given material thickness, different optimum beam and process parameters can be determined empirically, leading to a dross-free cut for high tool travel speeds. Realising sharp 90-degree corners, dross formation is observed and leads to a deteriorated cutting quality. With reorientation at small radii, the speed-dependent change in the cutting process is superimposed by the existing beam to nozzle misalignment and contributes to the stability of a cut. The feature radius R on the stability of the cutting process is being determined by reducing feature radius R. In this paper, cutting of different radii for different sized conventional nozzles is considered and analysed. Based on cutting quality evaluation, fine feature cutting is defined by discussing thickness-dependent finest cutting feature for a given gas dynamic input.

  4. Rational cutting height for large cutting height fully mechanized top-coal caving

    Institute of Scientific and Technical Information of China (English)

    Huang Bingxiang; Li Hongtao; Liu Changyou; Xing Shijun; Xue Weichao

    2011-01-01

    Large cutting height fully mechanized top-coal caving is a new mining method that improves recovery ratio and single-pass production.It also allows safe and efficient mining.A rational cutting height is one key parameter of this technique.Numerical simulation and a granular-media model experiment were used to analyze the effect of cutting height on the rock pressure of a fully mechanized top-coal caving work face.The recovery ratio was also studied.As the cutting height increases the top-coal thickness is reduced.Changing the ratio of cutting to drawing height intensifies the face pressure and the top-coal shattering.A maximum cutting height exists under a given set of conditions due to issues with surrounding rock-mass control.An increase in cutting height makes the top-coal cave better and the recovery ratio when drawing top-coal is then improved.A method of adjusting the face rock pressure is presented.Changing the cutting to drawing height ratio is the technique used to control face rock pressure.The recovery ratio when cutting coal exceeds that when caving top-coal so the face recovery ratio may be improved by over sizing the cutting height and increasing the top-coal drawing ratio.An optimum ratio of cutting to drawing height exists that maximizes the face recovery ratio.A rational cutting height is determined by comprehensively considering the surrounding rock-mass control and the recovery ratio.At the same time increasing the cutting height can improve single pass mining during fully mechanized top-coal caving.

  5. Laser Cutting of Leather: Tool for Industry or Designers?

    Science.gov (United States)

    Stepanov, Alexander; Manninen, Matti; Pärnänen, Inni; Hirvimäki, Marika; Salminen, Antti

    Currently technologies which are applied for leather cutting include slitting knifes, die press techniques and manual cutting. Use of laser technology has grown significantly during recent years due to number of advantages over conventional cutting methods; flexibility, high production speed, possibility to cut complex geometries, easier cutting of customized parts, and less leftovers of leather makes laser cutting more and more economically attractive to apply for leather cutting. Laser technology provides advantages in cutting complex geometries, stable cutting quality and possibility to utilize leather material in economically best way. Constant quality is important in industrial processes and laser technology fulfills this requirement: properly chosen laser cutting parameters provides identical cuts. Additionally, laser technology is very flexible in terms of geometries: complex geometries, individual designs, prototypes and small scale products can be manufactured by laser cutting. Variety of products, which needed to be cut in small volumes, is also the application where laser cutting can be more beneficial due to possibility to change production from one product to another only by changing geometry without a need to change cutting tool. Disadvantages of laser processing include high initial investment costs and some running costs due to maintenance and required gas supply for the laser. Higher level of operator's expertise is required due to more complicated machinery in case of laser cutting. This study investigates advantages and disadvantages of laser cutting in different areas of application and provides comparison between laser cutting and mechanical cutting of leather.

  6. Combining SSH and cDNA microarrays for rapid identification of differentially expressed genes.

    Science.gov (United States)

    Yang, G P; Ross, D T; Kuang, W W; Brown, P O; Weigel, R J

    1999-03-15

    Comparing patterns of gene expression in cell lines and tissues has important applications in a variety of biological systems. In this study we have examined whether the emerging technology of cDNA microarrays will allow a high throughput analysis of expression of cDNA clones generated by suppression subtractive hybridization (SSH). A set of cDNA clones including 332 SSH inserts amplified by PCR was arrayed using robotic printing. The cDNA arrays were hybridized with fluorescent labeled probes prepared from RNA from ER-positive (MCF7 and T47D) and ER-negative (MDA-MB-231 and HBL-100) breast cancer cell lines. Ten clones were identified that were over-expressed by at least a factor of five in the ER-positive cell lines. Northern blot analysis confirmed over-expression of these 10 cDNAs. Sequence analysis identified four of these clones as cytokeratin 19, GATA-3, CD24 and glutathione-S-transferase mu-3. Of the remaining six cDNA clones, four clones matched EST sequences from two different genes and two clones were novel sequences. Flow cytometry and immunofluorescence confirmed that CD24 protein was over-expressed in the ER-positive cell lines. We conclude that SSH and microarray technology can be successfully applied to identify differentially expressed genes. This approach allowed the identification of differentially expressed genes without the need to obtain previously cloned cDNAs.

  7. Cloning and expression of a cDNA for mouse prostaglandin E receptor EP2 subtype.

    Science.gov (United States)

    Honda, A; Sugimoto, Y; Namba, T; Watabe, A; Irie, A; Negishi, M; Narumiya, S; Ichikawa, A

    1993-04-15

    A functional cDNA clone encoding mouse EP2 subtype of prostaglandin (PG) E receptor was isolated from a mouse cDNA library by cross-hybridization with the mouse EP3 subtype PGE receptor cDNA. The mouse EP2 receptor consists of 513 amino acid residues with putative seven-transmembrane domains. In contrast to EP3 receptor, this receptor possesses long third intracellular loop and carboxyl-terminal tail. [3H] PGE2 specifically bound to the membrane of mammalian COS cells transfected with the cDNA. The binding to the membrane was displaced with unlabeled PG in the order of PGE2 = PGE1 > iloprost > or = PGF2 alpha > or = PGD2. The binding was also inhibited by misoprostol, an EP2 and EP3 agonist, but not by sulprostone, an EP1 and EP3 agonist, and SC-19220, an EP1 antagonist. PGE2 markedly increased cAMP level in COS cells transfected with the cDNA. These results suggest that this receptor is EP2 subtype. Northern blot analysis demonstrated that the EP2 mRNA is widely expressed in various tissues, the abundant expression being observed in ileum, thymus, and mastocytoma P-815 cells.

  8. Molecular cloning and nucleotide sequence of cDNA for human liver arginase

    Energy Technology Data Exchange (ETDEWEB)

    Haraguchi, Y.; Takiguchi, M.; Amaya, Y.; Kawamoto, S.; Matsuda, I.; Mori, M.

    1987-01-01

    Arginase (EC3.5.3.1) catalyzes the last step of the urea cycle in the liver of ureotelic animals. Inherited deficiency of the enzyme results in argininemia, an autosomal recessive disorder characterized by hyperammonemia. To facilitate investigation of the enzyme and gene structures and to elucidate the nature of the mutation in argininemia, the authors isolated cDNA clones for human liver arginase. Oligo(dT)-primed and random primer human liver cDNA libraries in lambda gt11 were screened using isolated rat arginase cDNA as a probe. Two of the positive clones, designated lambda hARG6 and lambda hARG109, contained an overlapping cDNA sequence with an open reading frame encoding a polypeptide of 322 amino acid residues (predicted M/sub r/, 34,732), a 5'-untranslated sequence of 56 base pairs, a 3'-untranslated sequence of 423 base pairs, and a poly(A) segment. Arginase activity was detected in Escherichia coli cells transformed with the plasmid carrying lambda hARG6 cDNA insert. RNA gel blot analysis of human liver RNA showed a single mRNA of 1.6 kilobases. The predicted amino acid sequence of human liver arginase is 87% and 41% identical with those of the rat liver and yeast enzymes, respectively. There are several highly conserved segments among the human, rat, and yeast enzymes.

  9. Cloning and sequencing of complete -crystallin cDNA from embryonic lens of Crocodylus palustris

    Indian Academy of Sciences (India)

    Raman Agrawal; Reena Chandrashekhar; Anurag Kumar Mishra; Jetty Ramadevi; Yogendra Sharma; Ramesh K Aggarwal

    2002-06-01

    -Crystallin is a taxon-specific structural protein found in eye lenses. We present here the cloning and sequencing of complete -crystallin cDNA from the embryonic lens of Crocodylus palustris and establish it to be identical to the -enolase gene from non-lenticular tissues. Quantitatively, the -crystallin was found to be the least abundant crystallin of the crocodilian embryonic lenses. Crocodile -crystallin cDNA was isolated by RT-PCR using primers designed from the only other reported sequence from duck and completed by 5′- and 3′-rapid amplification of cDNA ends (RACE) using crocodile gene specific primers designed in the study. The complete -crystallin cDNA of crocodile comprises 1305 bp long ORF and 92 and 409 bp long untranslated 5′-and 3′-ends respectively. Further, it was found to be identical to its putative counterpart enzyme -enolase, from brain, heart and gonad, suggesting both to be the product of the same gene. The study thus provides the first report on cDNA sequence of -crystallin from a reptilian species and also re-confirms it to be an example of the phenomenon of gene sharing as was demonstrated earlier in the case of peking duck. Moreover, the gene lineage reconstruction analysis helps our understanding of the evolution of crocodilians and avian species.

  10. Cloning and characterization of a cDNA encoding human differentiation antigen 5D4

    Institute of Scientific and Technical Information of China (English)

    马凤蓉; 朱立平; 汪燚; 赵方萄; 史耕先; 李波; 李国燕; 张淑珍; 王讯

    2000-01-01

    A 1 846 bp cDNA is isolated from a human tonsil cell λgt 11 cDNA library (ATCC No. 37546) with mAb 5D4 reactive strongly with human B cell line 3D5, but weakly with human B cell line Daudi and human T cell line Jurkat as a probe. RT-PCR also shows a strong reaction in 3D5 cell and a weak reaction in Daudi and Jurkat cell for 5D4 mRNA. There is an open reading frame from 88 to 1 209 bp in 5D4 cDNA encoding a 374 AA protein. Both the Northern blot analysis and the two consecutive stop codens before start coden demonstrate that the cDNA is a full-length cDNA. Secondary structure prediction suggests that there are a region from 295 to 334 AA in the protein with strong hydrophobicity and a transmembrane helix region with high score from 313 to 334 AA with an orientation from the inside to the outside of the cell.

  11. cDNA cloning, sequence analysis, and chromosomal localization of the gene for human carnitine palmitoyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Finocchiaro, G.; Taroni, F.; Martin, A.L.; Colombo, I.; Tarelli, G.T.; DiDonato, S. (Istituto Nazionale Neurologico C. Besta, Milan (Italy)); Rocchi, M. (Istituto G. Gaslini, Genoa (Italy))

    1991-01-15

    The authors have cloned and sequenced a cDNA encoding human liver carnitine palmitoyltransferase an inner mitochondrial membrane enzyme that plays a major role in the fatty acid oxidation pathway. Mixed oligonucleotide primers whose sequences were deduced from one tryptic peptide obtained from purified CPTase were used in a polymerase chain reaction, allowing the amplification of a 0.12-kilobase fragment of human genomic DNA encoding such a peptide. A 60-base-pair (bp) oligonucleotide synthesized on the basis of the sequence from this fragment was used for the screening of a cDNA library from human liver and hybridized to a cDNA insert of 2255 bp. This cDNA contains an open reading frame of 1974 bp that encodes a protein of 658 amino acid residues including 25 residues of an NH{sub 2}-terminal leader peptide. The assignment of this open reading frame to human liver CPTase is confirmed by matches to seven different amino acid sequences of tryptic peptides derived from pure human CPTase and by the 82.2% homology with the amino acid sequence of rat CPTase. The NH{sub 2}-terminal region of CPTase contains a leucine-proline motif that is shared by carnitine acetyl- and octanoyltransferases and by choline acetyltransferase. The gene encoding CPTase was assigned to human chromosome 1, region 1q12-1pter, by hybridization of CPTase cDNA with a DNA panel of 19 human-hanster somatic cell hybrids.

  12. Study on Wusan Granule Anti-tumor Related Target Gene Screened by Cdna Microarray

    Institute of Scientific and Technical Information of China (English)

    YOU Zi-li; SHI Jin-ping; CHEN Hai-hong

    2006-01-01

    To screen Wusan Granule anti-tumor related target gene using cDNA microarray technique, both mRNA from Lewis lung carcinoma tissues treated by Wusan Granule and untreated control are reversibly transcribed to prepare cDNA probes which are labeled by Cy5 and Cy3. Then, the probes are hybridized to the mice cDNA microarray type MGEC-20S. After hybridization, the cDNA microarray is scanned by ScanArray 3 000 scanner and the data is analyzed by ImaGene 3 software to screen the differentially expressed genes. There are 45 differentially expressed genes including 18 known genes and 27 unknown genes between the two groups, and among them, 20 elevated genes and 25 reduced genes are identified. Additionally, the genes related to invasion and metastasis of malignant carcinomas are down-regulated and the genes related to apoptosis are up-regulated. The cDNA microarray technique is a high-throughput approach to screen the Wusan Granule anti-tumor related target genes, which allow us to explore the molecular biological mechanism on a genomic scale.

  13. Construction of cDNA Library from NPC Tissue and Screening of Antigenic Genes

    Institute of Scientific and Technical Information of China (English)

    Jun Shu; Xiaojuan He; Guancheng Li

    2006-01-01

    To construct cDNA library of nasopharyngeal carcinoma (NPC) and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX (serological identification of antigen by recombinant cDNA expression library). Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64×106 recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31,S100 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.

  14. Thermoelectrical numerical model of electrosurgical rf cutting

    Science.gov (United States)

    Protsenko, Dmitry E.; Pearce, John A.

    2001-06-01

    We developed a 3D thermo-electrical model of RF tissue cutting that takes into account variations in electrical and thermal properties with temperature and water content, dynamics of water evaporation and thermal and electrical processes at the tissue-scalpel interface. The model predicts measurable parameters of the electric circuit (tissue impedance, ESU output RMS voltage and current) and tissue cutting rate. Results of numerical simulations suggest that high circuit impedance during electrosurgical cutting can result not only from tissue dehydration but from the configuration of the electric field as well. It appears that the area of tissue-scalpel electric contact is significantly smaller than the area of the scalpel itself but is large enough to rule out electric sparks as a major mechanism of electrosurgical cutting.

  15. New opportunities for cutting with lasers.

    Science.gov (United States)

    Renner, Thomas; Mairhörmann, Dieter

    2004-01-01

    Laser microcutting is a leading technology for the manufacture of surgical instruments and medical implants. This article assesses its capabilities for stents and surgical-needle manufacture and for other tube-cutting applications that require microscaled designs.

  16. Cuts and coproducts of massive triangle diagrams

    Energy Technology Data Exchange (ETDEWEB)

    Abreu, Samuel [Higgs Centre for Theoretical Physics, School of Physics and Astronomy,The University of Edinburgh,Mayfield Road, Edinburgh EH9 3JZ, Scotland (United Kingdom); Institut de Physique Théorique, Université Paris Saclay, CEA, CNRS,Orme des Merisiers, F-91191 Gif-sur-Yvette (France); Britto, Ruth [Institut de Physique Théorique, Université Paris Saclay, CEA, CNRS,Orme des Merisiers, F-91191 Gif-sur-Yvette (France); School of Mathematics, Trinity College,College Green, Dublin 2 (Ireland); Hamilton Mathematical Institute, Trinity College,College Green, Dublin 2 (Ireland); Grönqvist, Hanna [Institut de Physique Théorique, Université Paris Saclay, CEA, CNRS,Orme des Merisiers, F-91191 Gif-sur-Yvette (France)

    2015-07-21

    Relations between multiple unitarity cuts and coproducts of Feynman integrals are extended to allow for internal masses. These masses introduce new branch cuts, whose discontinuities can be derived by placing single propagators on shell and identified as particular entries of the coproduct. First entries of the coproduct are then seen to include mass invariants alone, as well as threshold corrections for external momentum channels. As in the massless case, the original integral can possibly be recovered from its cuts by starting with the known part of the coproduct and imposing integrability contraints. We formulate precise rules for cuts of diagrams, and we gather evidence for the relations to coproducts through a detailed study of one-loop triangle integrals with various combinations of external and internal masses.

  17. Oil Refining Giants to Cut Output

    Institute of Scientific and Technical Information of China (English)

    Zhang Dingmin

    2002-01-01

    @@ China's refined oil product output rose slightly in the first five months of this year on strong demand,but the two biggest domestic producers plan to cut production rates in an attempt to bolster the sluggish price.

  18. On Triple-Cut of Scattering Amplitudes

    CERN Document Server

    Mastrolia, Pierpaolo

    2007-01-01

    It is analysed the triple-cut of one-loop amplitudes in dimensional regularisation within spinor-helicity representation. The triple-cut is defined as a difference of two double-cuts with the same particle content, and a same propagator carrying, respectively, causal and anti-causal prescription in each of the two cuts. That turns out into an effective tool for extracting the coefficients of the three-point functions (and higher-point ones) from one-loop-amplitudes. The phase-space integration is oversimplified by using residues theorem to perform the integration over the spinor variables, via the holomorphic anomaly, and a trivial integration on the Feynman parameter. The results are valid for arbitrary values of dimensions.

  19. Correlation of cutting fluid performance in different machining operations

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo; Belluco, Walter

    2001-01-01

    An analysis of cutting fluid performance in different metal cutting operations is presented, based on experimental investigations in which type of operation, performance criteria, work material, and fluid type are considered. Cutting fluid performance was evaluated in turning, drilling, reaming...

  20. Molecular cloning and characterization of a cDNA encoding the N-acetyl-beta-D-glucosaminidase homologue of Paracoccidioides brasiliensis.

    Science.gov (United States)

    Santos, Mônica O; Pereira, Maristela; Felipe, Maria Sueli S; Jesuino, Rosalia Santos A; Ulhoa, Cirano J; Soares, Renata de Bastos A; Soares, Celia Maria de A

    2004-06-01

    A cDNA encoding the N-acetyl-beta-D-glucosaminidase (NAG) protein of Paracoccidioides brasiliensis, Pb NAG1, was cloned and characterized. The 2663-nucleotide sequence of the cDNA consisted of a single open reading frame encoding a protein with a predicted molecular mass of 64.73 kDa and an isoeletric point of 6.35. The predicted protein includes a putative 30-amino-acid signal peptide. The protein as a whole shares considerable sequence similarity with 'classic' NAG. The primary sequence of Pb NAG1 was used to infer phylogenetic relationships. The amino acid sequence of Pb NAG1 has 45, 31 and 30% identity, respectively, with homologous sequences from Trichoderma harzianum, Aspergillus nidulans and Candida albicans. In particular, striking homology was observed with the active site regions of the glycosyl hydrolase group of proteins (family 20). The expected active site consensus motif G X D E and catalytic Asp and Glu residues at positions 373 and 374 were found, reinforcing that Pb NAG1 belongs to glycosyl hydrolase family 20. The nucleotide sequence of Pb nag1 and its flanking regions have been deposited, along with the amino acid sequence of the deduced protein, in GenBank under accession number AF419158.

  1. A new set of ESTs and cDNA clones from full-length and normalized libraries for gene discovery and functional characterization in citrus

    Directory of Open Access Journals (Sweden)

    Alamar Santiago

    2009-09-01

    EST collection denotes an important step towards the identification of all genes in the citrus genome. Furthermore, public availability of the cDNA clones generated in this study, and not only their sequence, enables testing of the biological function of the genes represented in the collection. Expression of the citrus SEP3 homologue, CitrSEP, in Arabidopsis results in early flowering, along with other phenotypes resembling the over-expression of the Arabidopsis SEPALLATA genes. Our findings suggest that the members of the SEP gene family play similar roles in these quite distant plant species.

  2. Link between chips and cutting moments evolution

    CERN Document Server

    Cahuc, Olivier; Gérard, Alain; 10.4028/WWW.scientific.net/AMR.423.89

    2012-01-01

    The better understanding of the material cutting process has been shown with the benefit of the forces and moments measurement since some years ago. In paper, simultaneous six mechanical components and chip orientation measurements were realized during turning tests. During these tests, the influence of the depth of cut or feed rate has been observed and a link between the chip orientation and the moment vector orientation or the central axis characteristics has been shown.

  3. From Dimensional to Cut-Off Regularization

    CERN Document Server

    Dillig, M

    2006-01-01

    We extent the standard approach of dimensional regularization of Feynman diagrams: we replace the transition to lower dimensions by a 'natural' cut-off regulator. Introducing an external regulator of mass Lambda^(2e), we regain in the limit e -> 0 and e > 0 the results of dimensional and cut-off regularization, respectively. We demonstrate the versatility and adequacy of the different regularization schemes for practical examples (such as non covariant regularization, the axial anomaly or regularization in effective field theories).

  4. Rooting of carnation cuttings: The auxin signal

    OpenAIRE

    Acosta, Manuel; Oliveros-Valenzuela, M Rocío; Nicolás, Carlos; Sánchez-Bravo, José

    2009-01-01

    The rooting of stem cuttings is a common vegetative propagation practice in many ornamental species. Among other signals, auxin polarly transported through the stem plays a key role in the formation and growth of adventitious roots. Unlike in other plant species, auxin from mature leaves plays a decisive role in the rooting of carnation (Dianthus caryophyllus. L) cuttings. The gene DcAUX1, which codifies an auxin influx carrier involved in polar auxin transport, has now been cloned and charac...

  5. Cut Size Statistics of Graph Bisection Heuristics

    OpenAIRE

    Schreiber, G. R.; Martin, O. C.

    1998-01-01

    We investigate the statistical properties of cut sizes generated by heuristic algorithms which solve approximately the graph bisection problem. On an ensemble of sparse random graphs, we find empirically that the distribution of the cut sizes found by ``local'' algorithms becomes peaked as the number of vertices in the graphs becomes large. Evidence is given that this distribution tends towards a Gaussian whose mean and variance scales linearly with the number of vertices of the graphs. Given...

  6. Geometric Modelling by Recursively Cutting Vertices

    Institute of Scientific and Technical Information of China (English)

    吕伟; 梁友栋; 等

    1989-01-01

    In this paper,a new method for curve and surface modelling is introduced which generates curves and surfaces by recursively cutting and grinding polygons and polyhedra.It is a generalization of the existing corner-cutting methods.A lot of properties,such as geometric continuity,representation,shape-preserving,and the algorithm are studied which show that such curves and surfaces are suitable for geometric designs in CAD,computer graphics and their application fields.

  7. Cutting Symmetrical Recesses In Soft Ceramic Tiles

    Science.gov (United States)

    Nesotas, Tony C.; Tyler, Brent

    1989-01-01

    Simple tool cuts hemispherical recesses in soft ceramic tiles. Designed to expose wires of thermocouples embedded in tiles without damaging leads. Creates neat, precise holes around wires. End mill includes axial hole to accommodate thermocouple wires embedded in material to be cut. Wires pass into hole without being bent or broken. Dimensions in inches. Used in place of such tools as dental picks, tweezers, spatulas, and putty knives.

  8. Alternative Metal Hot Cutting Operations for Opacity

    Science.gov (United States)

    2014-11-01

    testing was conducted on 9/32” ordinary strength steel bulkhead section from the ex-CGN-9 and, if possible, 2” high yield strength steel ( HY80 ...recorded for both the alternative fuel and propane. Test cuts were on 9/32” steel surface ship sections with additional demonstration cuts performed on...thicker surface ship sections and 2” high yield strength steel submarine pressure hull as the pieces became available. The results from the

  9. LASER CUTTING MACHINES FOR 3-D THIN SHEET PARTS

    Directory of Open Access Journals (Sweden)

    Miroslav RADOVANOVIC

    2012-11-01

    Full Text Available Laser cutting machines are used for precise contour cutting thin sheet. In industrial application nowadays various types and construction of laser cutting machines can be met. For contour cutting 3-D thin sheet parts laser cutting machines with rotation movements and laser robots are used. Laser generates the light beam, that presents a tool in working process. Application of laser cutting machines made possible good quality of products, flexibility of production and enlargement of economy

  10. New law to cut 1986 budgets

    Science.gov (United States)

    Katzoff, Judith A.

    The National Aeronautics and Space Administration (NASA), the National Oceanic and Atmospheric Administration (NOAA), the National Science Foundation (NSF), and the U.S. Geological Survey (USGS) are among the agencies whose budgets are likely to be cut 4.3% on March 1, 1986, when the Balanced Budget and Emergency Deficit Control Act of 1985 makes its first impact.Under the law, the cuts will be implemented across the board by an order from the president. Unless Congress passes legislation to make its own cuts sometime between now and March 1, these cuts will automatically take an equal percentage from every nondefense program, project, and activity not protected from “sequestration” of funds and will automatically take a slightly higher percentage from the unprotected portions of the defense budget. Individual agencies could circumvent the across-the-board cuts by formulating their own plans to distribute their share of the reductions; however, these plans would need congressional approval. These cuts will affect budgets for the federal government in fiscal year (FY) 1986, which began on October 1, 1985. Unless revenues increase substantially, additional and mone severe belt-tightening is in store for FY 1987 and beyond.

  11. Demonstrating diamond wire cutting of the TFTR

    Energy Technology Data Exchange (ETDEWEB)

    Rule, K.; Perry, E.; Larson, S.; Viola, M. [and others

    2000-02-24

    The Tokamak Fusion Test Reactor (TFTR) ceased operation in April 1997 and decommissioning commenced in October 1999. The deuterium-tritium fusion experiments resulted in contaminating the vacuum vessel with tritium and activating the materials with 14 Mev neutrons. The total tritium content within the vessel is in excess of 7,000 Curies while dose rates approach 50 mRem/hr. These radiological hazards along with the size of the Tokamak (100 cubic meters) present a unique and challenging task for dismantling. Plasma arc cutting is the current baseline technology for the dismantlement of fission reactors. This technology is typically used because of its faster cutting times. Alternatively, an innovative approach for dismantlement of the TFTR is the use of diamond wire cutting technology. Recent improvements in diamond wire technology have allowed the cutting of carbon steel components such as pipe, plate, and tube bundles in heat exchangers. Some expected benefits of this technology include: significantly reduction in airborne contaminates, reduced personnel exposure, a reduced risk of spread of tritium contamination, and reduced overall costs as compared to using plasma arc cutting. This paper will provide detailed results of the diamond wire cutting demonstration that was completed in September of 1999, on a mock-up of this complex reactor. The results will identify cost, safety, industrial and engineering parameters, and the related performance of each situation.

  12. Plasma arc cutting technology: simulation and experiments

    Energy Technology Data Exchange (ETDEWEB)

    Cantoro, G; Colombo, V; Concetti, A; Ghedini, E; Sanibondi, P; Zinzani, F; Rotundo, F [Department of Mechanical Engineering (D.I.E.M.) and Research Center for Applied Mathematics (C.I.R.A.M.), Alma Mater Studiorum-Universita di Bologna, Via Saragozza 8, 40123 Bologna (Italy); Dallavalle, S; Vancini, M, E-mail: emanuele.ghedini@unibo.it [Cebora S.p.A., Via Andrea Costa 24, 40057 Cadriano di Granarolo (Italy)

    2011-01-01

    Transferred arc plasma torches are widely used in industrial processes for cutting of metallic materials because of their ability to cut a wide range of metals with very high productivity. The process is characterized by a transferred electric arc established between an electrode inside the torch (the cathode) and another electrode, the metallic workpiece to be cut (the anode). In order to obtain a high quality cut and a high productivity, the plasma jet must be as collimated as possible and must have the higher achievable power density. Plasma modelling and numerical simulation can be very useful tools for the designing and optimizing these devices, but research is still in the making for finding a link between simulation of the plasma arc and a consistent prevision of cut quality. Numerical modelling of the behaviour of different types of transferred arc dual gas plasma torches can give an insight on the physical reasons for the industrial success of various design and process solutions that have appeared over the last years. Diagnostics based on high speed imaging and Schlieren photography can play an important role for investigating piercing, dross generation, pilot arcing and anode attachment location. Also, the behaviour of hafnium cathodes at high current levels at the beginning of their service life can been experimentally investigated, with the final aim of understanding the phenomena that take place during those initial piercing and cutting phases and optimizing the initial shape of the surface of the emissive insert exposed to plasma atmosphere.

  13. Ion implantation of superhard ceramic cutting tools

    Science.gov (United States)

    Chou, Y. Kevin; Liu, Jie

    2004-08-01

    Despite numerous reports of tool life increase by ion implantation in machining operations, ion implantation applications of cutting tools remain limited, especially for ceramic tools. Mechanisms of tool-life improvement by implantation are not clearly established due to complexity of both implantation and tool-wear processes. In an attempt to improve performance of cubic boron nitride (CBN) tools for hard machining by ion implantation, a literature survey of ion-implanted cutting tools was carried out with a focus on mechanisms of tool-wear reduction by ion implantation. Implantation and machining experiments were then conducted to investigate implantation effects on CBN tools in hard machining. A batch of CBN tools was implanted with nitrogen ions at 150 keV and 2.5×1017 ions/cm2 and further used to cut 61 HRc AISI 52100 steel at different conditions. Results show that ion implantation has strong effects on partsurface finish, moderate effect on cutting forces, but an insignificant impact on tool wear. Friction coefficients, estimated from measured cutting forces, are possibly reduced by ion implantation, which may improve surface finish. However, surprisingly, 2-D orthogonal cutting to evaluate tribological loading in hard machining showed no difference on contact stresses and friction coefficients between implanted and nonimplanted CBN tools.

  14. Quantum Max-flow/Min-cut

    Science.gov (United States)

    Cui, Shawn X.; Freedman, Michael H.; Sattath, Or; Stong, Richard; Minton, Greg

    2016-06-01

    The classical max-flow min-cut theorem describes transport through certain idealized classical networks. We consider the quantum analog for tensor networks. By associating an integral capacity to each edge and a tensor to each vertex in a flow network, we can also interpret it as a tensor network and, more specifically, as a linear map from the input space to the output space. The quantum max-flow is defined to be the maximal rank of this linear map over all choices of tensors. The quantum min-cut is defined to be the minimum product of the capacities of edges over all cuts of the tensor network. We show that unlike the classical case, the quantum max-flow=min-cut conjecture is not true in general. Under certain conditions, e.g., when the capacity on each edge is some power of a fixed integer, the quantum max-flow is proved to equal the quantum min-cut. However, concrete examples are also provided where the equality does not hold. We also found connections of quantum max-flow/min-cut with entropy of entanglement and the quantum satisfiability problem. We speculate that the phenomena revealed may be of interest both in spin systems in condensed matter and in quantum gravity.

  15. Molecular cloning and sequencing of a cDNA encoding partial putative molt-inhibiting hormone from Penaeus chinensis

    Science.gov (United States)

    Wang, Zai-Zhao; Xiang, Jian-Hai

    2002-09-01

    Total RNA was extracted from eyestalks of shrimp Penaeus chinensis. Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase-polymerase chain reaction (RT-PCR) was initiated using eyestalk cDNA and degenerate primers designed from the amino acid sequence of molt-inhibiting hormone from shrimp Penaeus japonicus. A specific cDNA was obtained and cloned into a T vector for sequencing. The cDNA consisted of 201 base pairs and encoding for a peptide of 67 amino acid residues. The peptide of P. chinensis had the highest identity with molt-inhibiting hormones of P. japonicus. The cDNA could be a partial gene of molt-inhibiting hormones from P. chinensis. This paper reports for the first time cDNA encoding for neuropeptide of P. chinensis.

  16. MOLECULAR CLONING AND SEQUENCING OF A cDNA ENCODING PARTIAL PUTATIVE MOLT-INHIBITING HORMONE FROM PENAEUS CHINENSIS

    Institute of Scientific and Technical Information of China (English)

    王在照; 相建海

    2002-01-01

    Total RNA was extracted from eyestalks of shrimp Penaeus chinensis. Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase-polymer ase chain reaction (RT-PCR) was initiated using eyestalk cDNA and degenerate primers designed from the amino acid sequence of molt-inhibiting hormone from shrimp Penaeus japonicus. A s pecific cDNA was obtained and cloned into a T vector for sequencing. The cDNA consisted of 201 ba se pairs and encoding for a peptide of 67 amino acid residues. The peptide of P. chinensis had the highest identity with molt-inhibiting hormones of P. japonicus. The cDNA could be a partial gene of molt-inhibiting hormones from P. chinensis. This paper reports for the first time cDNA encoding for neuropeptide of P. chinensis.

  17. Construction of a Plant Transformation-ready Expression cDNA Library for Thellungiella halophila Using Recombination Cloning

    Institute of Scientific and Technical Information of China (English)

    Wan-Song Ni; Zhi-Yong Lei; Xi Chen; David J. Oliver; Cheng-Bin Xiang

    2007-01-01

    Salt cress (Thellungiella halophila), a close relative of the model plant Arabidopsis thaliana L., is an extremophile that is adapted to harsh saline environments. To mine salt-tolerance genes from this species, we constructed an entry cDNA library from the salt cress plant treated with salt-stress by using a modified cDNA synthesis and an improved recombinationassisted cDNA library construction method that is completely free of manipulations involving restriction enzymes and DNA ligase. This cDNA library construction procedure is significantly simplified and the quality of the cDNA library is improved. This entry cDNA library was subsequently shuttled into the destination binary vector pCB406 designed for plant transformation and expression via recombination-assisted cloning. The library is plant transformation ready and is used to transform Arabidopsis on a large scale in order to create a large collection of transgenic lines for functional gene mining.

  18. MOLECULAR CLONING AND SEQUENCING OF A cDNA ENCODING PARTIAL PUTATIVE MOLT-INHIBITING HORMONE FROM PENAEUS CHINENSIS

    Institute of Scientific and Technical Information of China (English)

    王在照; 相建海

    2002-01-01

    Total RNA was extracted from eyestalks of shrimp Penaeue chinensis. Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase-polymerase chain reaction (RT-PCR) was initiated using eyestalk cDNA and degenerate primers designed from the amino acid sequence of molt-inhibiting hormone from shrimp Penaeus japonicus. A specific cDNA was obtained and cloned into a T vector for sequencing. The cDNA consisted of 201 base pairs and encoding for a peptide of 67 amino acid residues. The peptide of P. chinensis had the highest identity with molt-inhibiting hormones of P. japonicus. The cDNA could be a partial gene of molt-inhibiting hormones from P. chinensis. This paper reports for the first time cDNA encoding for neuropeptide of P. chinensis.

  19. EXPERIMENTAL ESTIMATION OF TOOL WEAR AND CUTTING TEMPERATURES IN MQL USING CUTTING FLUIDS WITH CNT INCLUSION

    Directory of Open Access Journals (Sweden)

    S.NARAYANA RAO,

    2011-04-01

    Full Text Available Machining often witnesses frequent interruption caused by friction and heat. Cutting fluids are being used in machining for centuries to counter the effects of friction and temperatures. However, because of many disadvantages, MQL has emerged. MQL demands fluids with high performance. This work tries to estimate tool wear and cutting temperatures while using cutting fluids, prepared with carbon nano tube (CNT inclusion.

  20. Identification of a Herbicide Safener AD-67 Inducible cDNA in Rice

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A herbicide safener AD-67 inducible cDNA was identified in an indica rice variety 9311 by mRNA differential display. The transcript was increased 6 h after sprayed with the safener solution, and 4 days later, the expression still could be detected. The fragment was recycled from the poly-gel and sequenced, and homologous analysis revealed the cDNA was 100% identical to some ESTs and cDNAs in rice database, and the amino acid sequence was 60-84% homologous to those of the Yippee genes in several eukaryotes. The fragment was extended to the whole long cDNA, and thus a primer pair was designed. RT-PCR analysis for the designed primer supported the induction result.

  1. Design and Screening of M13 Phage Display cDNA Libraries

    Directory of Open Access Journals (Sweden)

    Yuliya Georgieva

    2011-02-01

    Full Text Available The last decade has seen a steady increase in screening of cDNA expression product libraries displayed on the surface of filamentous bacteriophage. At the same time, the range of applications extended from the identification of novel allergens over disease markers to protein-protein interaction studies. However, the generation and selection of cDNA phage display libraries is subjected to intrinsic biological limitations due to their complex nature and heterogeneity, as well as technical difficulties regarding protein presentation on the phage surface. Here, we review the latest developments in this field, discuss a number of strategies and improvements anticipated to overcome these challenges making cDNA and open reading frame (ORF libraries more readily accessible for phage display. Furthermore, future trends combining phage display with next generation sequencing (NGS will be presented.

  2. Isolation and Characterization of Phytoene Desaturase cDNA from Stigma of Crocus sativus

    Institute of Scientific and Technical Information of China (English)

    Bai Jie(白洁); Xu Ying; Tang Lin; Zeng Yu; Feng Yun; Wang Shenghua; Chen Fang

    2004-01-01

    Phytoene desaturase (PDS) has recently been identified as an important enzyme in carotenoid biosynthesis pathway. A cDNA clone encoding phytoene desaturase gene is isolated from stigma of saffron (Crocus sativus L.) using RT-PCR technique. Sequence analysis shows 83% similarity to Narcissus pseudonarcissus, 79% to Zea mays, 78% to Arabidopsis thaliana, 77% to Lycopersicon esculentum. A new full-length cDNA is obtained by 5'-RACE and 3' -RACE techniques. The cDNA is 2149bp long with an open reading frame of 1697bp, which encodes a polypeptide of 565 amino acids. Southern analysis shows that the PDS gene is a single copy in saffron. Northern blot analysis shows higher expression level of PDS gene in stigma and anther than in leaves and stem.

  3. cDNA macroarray for analysis of gene expression profiles in prostate cancer

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Background Early diagnosis and timely treatment are important for improving therapeutic efficiency of prostate cancer. DNA array is a new bio-technology for disease diagnosis. This study was conducted to diagnose prostate cancer with cDNA macroarray and analysis gene expression profiles of some selective genes in prostate cancer.Methods Total RNA was isolated from patients with prostate cancer and from normal people, and poly(A) RNA was further purified. Then it was analyzed for differentially expressed genes in prostate cancer and normal prostate by cDNA macroarray system.Results There were different expressions in the nine prostate-associated specific genes in prostate cancer as compared with normal prostate, in which, 7 were significantly upregulated and 2 were down-regulated.Conclusion As a diagnostic approach at molecular level, the cDNA macroarray is an effectively diagnostic method for prostate cancer.

  4. LD-RTPCR:\tA NEW METHOD FOR LABELLING TRACE cDNA MICROARRAY PROBE

    Institute of Scientific and Technical Information of China (English)

    范保星; 孙敬芬; 梁好; 王升启; 周平坤; 吴德昌

    2002-01-01

    Objective: To explore the usefulness of long distance reverse transcript combining linear amplification (LD-RTPCR) in labeling slight trace probe used for cDNA microarray. Methods: Total RNA from BEP2D cells was extracted and labeled by two different methods, LD-RTPCR with Cy3-dCTP as fluorescent dye and traditionally used RNA reverse transcript (RT) with Cy5-dCTP as fluorescent dye. Then, the probes labeled by two methods were mixed equally and hybridized with the cDNA microarray. Results: Scan and analysis of the microarray showed that the two methods labeled probes had consistent results. Conclusion: LD-RTPCR was proved useful for labeling cDNA microarray probe, especially for limited RNA material.

  5. Optimization of yeast surface-displayed cDNA library screening for low abundance targets.

    Science.gov (United States)

    Kim, Juh-Yung; Kim, Hyung Kyu; Jang, Hye Jeong; Kim, Eun-Kyung; Kim, Moon Kyu

    2015-04-01

    The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.

  6. First characterization of infectious cDNA clones of Olive mild mosaic virus

    Directory of Open Access Journals (Sweden)

    Joana M.S. CARDOSO

    2012-09-01

    Full Text Available Full-length cDNA clones of an Olive mild mosaic virus (OMMV isolate were constructed in order to find infectious cDNA clones. The sequencing of three individual full-length clones revealed some differences between them. In vitro transcription of these clones was performed and the effect of spontaneous mutations in the biological behaviour of the in vitro transcripts was evaluated by symptomatology, RNA accumulation and virus replication in inoculated plants. In vitro synthesized RNA from one of these clones was found to mimic the wild-type OMMV, making it useful in future studies on protein structure and function by site directed mutagenesis of individual genes. This is the first report on constructing full-length cDNA clones of OMMV from which infectious RNAs can be transcribed in vitro.

  7. Profiling gene expression patterns of nasopharyngeal carcinoma and normal nasopharynx tissues with cDNA microarray

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    5 μg of total RNAs from normal nasopharynx and nasopharyngeal carcinoma tissue have been labeled with α-32P-dCTP during reverse transcription. The synthesized cDNA probes have been hybridized to high-density cDNA microarray containing 5184 genes or expression sequence tags (ESTs). Then image analysis software has been applied to comparing their expression profiles. Results show that 187 ESTs were of density value above 200 in nasopharyngeal carcinoma tissue while there were 307 such ESTs in normal nasopharynx tissue; 38 ESTs were strongly expressed in nasopharynx, but weakly expressed in nasopharyngeal carcinoma; 48 ESTs were strongly expressed in nasopharyngeal carcinoma, but weakly expressed in normal nasopharynx. These results suggest that there may exist some new differentially expressed genes involved in nasopharyngeal carcinoma development. Furthermore, the results strongly indicate that high-density cDNA microarray is a powerful and efficient tool for large-scale screening differentially expressed genes.

  8. Regulation of human clotting factor IX cDNA expression in transgenic mice

    Institute of Scientific and Technical Information of China (English)

    胡以平; 邱信芳; 薛京伦; 刘祖洞

    1995-01-01

    To study the expression of human dotting factor IX cDNA in transgenic mice,Which is an es-sential work on gene therapy for hemophilia B,3 recombinant constructions containing different lengths ofhuman dotting factor IX cDNA have been introduced into the cultured cells.All of the recombinant constructionswere found to he expressed well in vitro.They were then microinjected into the male pronudei of the fertilizedmouse eggs respectively for generating trahsgenic mice.Unfortunately,none of them was expressed in any transgenicmice.These results show that the expression of the human clotting factor IX cDNA in the transgenic mice canbe determined by cis regulatory element(s).As compared With the results from other related works,it is sug-gested that the cis regulatory element(s)is resided in the 5’-end non-coding region.

  9. Laser cutting of various materials: Kerf width size analysis and life cycle assessment of cutting process

    Science.gov (United States)

    Yilbas, Bekir Sami; Shaukat, Mian Mobeen; Ashraf, Farhan

    2017-08-01

    Laser cutting of various materials including Ti-6Al-4V alloy, steel 304, Inconel 625, and alumina is carried out to assess the kerf width size variation along the cut section. The life cycle assessment is carried out to determine the environmental impact of the laser cutting in terms of the material waste during the cutting process. The kerf width size is formulated and predicted using the lump parameter analysis and it is measured from the experiments. The influence of laser output power and laser cutting speed on the kerf width size variation is analyzed using the analytical tools including scanning electron and optical microscopes. In the experiments, high pressure nitrogen assisting gas is used to prevent oxidation reactions in the cutting section. It is found that the kerf width size predicted from the lump parameter analysis agrees well with the experimental data. The kerf width size variation increases with increasing laser output power. However, this behavior reverses with increasing laser cutting speed. The life cycle assessment reveals that material selection for laser cutting is critical for the environmental protection point of view. Inconel 625 contributes the most to the environmental damages; however, recycling of the waste of the laser cutting reduces this contribution.

  10. Analysis of gene expression profile of aspermia using cDNA microarray

    Institute of Scientific and Technical Information of China (English)

    杨波; 高晓康; 王禾; 刘贺亮; 陈宝琦; 秦荣良; 康福霞; 邵国兴; 邵晨

    2003-01-01

    Objective: To identify the differential gene expression profiles between the normal and aspermia human testes utilizing cDNA microarray. Methods: cDNA probes were prepared by labeling mRNA of aspermia testes tissues with Cy5-dUTP and mRNA of normal testes tissues with Cy3-dUTP respectively through reverse transcription. The mixed cDNA probes were then hybridized with 4096 cDNA arrays (4096 unique human cDNA sequences), and the fluorescent signals were scanned by ScanArray 3000 scanner (General Scanning, Inc.). The values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated by ImaGene 3.0 software (BioDiscovery, Inc.). Differentially expressed genes were screened according to the criterion that the absolute value of natural logarithm of the ratio of Cy5-dUTP to Cy3-dUTP was greater-than 2.0 or less-than 0.5. A randomly chosen gene RAP1A was studied by in situ hybridization to evaluate the accuracy of the results. Results: 623 differential expressed genes related to aspermia were found. There were 303 up-expressed genes and 320 down-expressed genes. A distinct up-expressed gene RAP1A was confirmed by in situ hybridization. Conclusions: Screening the differential gene expression profiles between the normal and aspermia human testis by cDNA microarray can be used in the study of aspermia-related genes and the further research due to its properties, RAP1A may play some roles in the development and progression of aspermia.

  11. Establishment of Self-incompatibility Gene cDNA Microarray to Identify S-genotypes of Pyrus pyrifolia

    Directory of Open Access Journals (Sweden)

    Jiang Nan

    2015-11-01

    Full Text Available Based on the cDNA sequences from hyper variable (HV regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase cDNA probes were designed, and a cDNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 cDNA probes, including all specific cDNA sequences from the HV regions of the S-alleles. Using the cDNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the cDNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli’, ‘Xiuyu’, ‘Midu Yuli’, ‘Baimianli’, and ‘Deshengxiang’ were similar to the known genotypes of all tested cultivars. The S-RNase cDNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of cDNA microarrays has further improved the pear S-RNase detection platform.

  12. Family Privilege

    Science.gov (United States)

    Seita, John R.

    2014-01-01

    Family privilege is defined as "strengths and supports gained through primary caring relationships." A generation ago, the typical family included two parents and a bevy of kids living under one roof. Now, every variation of blended caregiving qualifies as family. But over the long arc of human history, a real family was a…

  13. Family Privilege

    Science.gov (United States)

    Seita, John R.

    2014-01-01

    Family privilege is defined as "strengths and supports gained through primary caring relationships." A generation ago, the typical family included two parents and a bevy of kids living under one roof. Now, every variation of blended caregiving qualifies as family. But over the long arc of human history, a real family was a…

  14. Cloning and screening of cDNA of Psilgramma menephorn allergen

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To construct a cDNA expression library of Psilgramma menephorn to screen its major allergen so as to provide the basis for producing recombinant allergen vaccine of Psilgramma menephorn. Methods Total RNA was extracted from the whole body of Psilgramma menephorn with Trizol and mRNA was purified with Oligo (dT) Spin-Column. And dscDNA was synthesized through reverse transcription. After blunting, the cDNA fragments were ligated with EcoRⅠ adapters. Then the cDNAs were digested by XhoⅠ, and the fra...

  15. Avoiding cross hybridization by choosing nonredundant targets on cDNA arrays

    DEFF Research Database (Denmark)

    Nielsen, Henrik Bjørn; Knudsen, Steen

    2002-01-01

    PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end.......PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end....

  16. Characterization of Expressed Sequence Tags From a Gallus gallus Pineal Gland cDNA Library

    OpenAIRE

    2005-01-01

    The pineal gland is the circadian oscillator in the chicken, regulating diverse functions ranging from egg laying to feeding. Here, we describe the isolation and characterization of expressed sequence tags (ESTs) isolated from a chicken pineal gland cDNA library. A total of 192 unique sequences were analysed and submitted to GenBank; 6% of the ESTs matched neither GenBank cDNA sequences nor the newly assembled chicken genomic DNA sequence, three ESTs aligned with sequences designated to be on...

  17. cDNA cloning of the beta subunit of teleost thyrotropin.

    OpenAIRE

    Ito, M.; Koide, Y.; Takamatsu, N; Kawauchi, H.; Shiba, T.

    1993-01-01

    cDNA clones encoding the beta subunit of thyrotropin (thyroid-stimulating hormone; TSH) were isolated from a cDNA library made from the pituitaries of immature rainbow trout and sequenced. The precursor of rainbow trout TSH beta consists of 147 aa, which can be cleaved into a signal peptide (20 aa) and a mature protein (127 aa) containing one potential N-glycosylation site and 12 cysteine residues. The protein showed highest homology with human TSH beta (51%) and lesser homology with human fo...

  18. Index of /data/medaka-full-length-cdna-db/20110331 [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Index of /data/medaka-full-length-cdna-db/20110331 Name Last modified Size Descript...ion Parent Directory - README.html 06-Sep-2012 16:47 13K medaka_full_length_c..> 26-Aug-2011 15:54 99M medaka_full_leng...th_c..> 26-Aug-2011 15:52 814K medaka_full_length_c..> 26-Aug-2011 15:52 8.7M Index of /data/medaka-full-length-cdna-db/20110331 ...

  19. Download - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project Download First of all, please read the license of this database. Data ...names and data descriptions are about the downloadable data in this page. They might not correspond to the c...f the data. # Data name File Simple search and download 1 README README_e.html - 2 5'-end sequences of buddi...ng yeast full-length cDNA clones and quality scores yeast_seq_qual.zip (59.9MB) Simple search and download 3...Downlaod via FTP Joomla SEF URLs by Artio About This Database Database Description Download License Update H

  20. Folate biosynthesis in higher plants. cDNA cloning, heterologous expression, and characterization of dihydroneopterin aldolases.

    Science.gov (United States)

    Goyer, Aymeric; Illarionova, Victoria; Roje, Sanja; Fischer, Markus; Bacher, Adelbert; Hanson, Andrew D

    2004-05-01

    Dihydroneopterin aldolase (EC 4.1.2.25) is one of the enzymes of folate synthesis that remains to be cloned and characterized from plants. This enzyme catalyzes conversion of 7,8-dihydroneopterin (DHN) to 6-hydroxymethyl-7,8-dihydropterin, and is encoded by the folB gene in Escherichia coli. The E. coli FolB protein also mediates epimerization of DHN to 7,8-dihydromonapterin. Searches of the Arabidopsis genome detected three genes encoding substantially diverged FolB homologs (AtFolB1-3, sharing 57%-73% identity), for which cDNAs were isolated. A fourth cDNA specifying a FolB-like protein (LeFolB1) was obtained from tomato (Lycopersicon esculentum) by reverse transcription-PCR. When overproduced in E. coli, recombinant AtFolB1, AtFolB2, and LeFolB1 proteins all had both dihydroneopterin aldolase and epimerase activities, and carried out the aldol cleavage reaction on the epimerization product, 7,8-dihydromonapterin, as well as on DHN. AtFolB3, however, could not be expressed in active form. Size exclusion chromatography indicated that the plant enzyme is an octamer, like the bacterial enzyme. Quantifying expression of the Arabidopsis genes by real-time reverse transcription-PCR showed that AtFolB1 and AtFolB2 messages occur at low levels throughout the plant, whereas the AtFolB3 mRNA was detected only in siliques and only with an extremely low abundance. Sequence comparisons and phylogenetic analysis of FolB homologs from 16 plants indicated that their N-terminal regions are highly variable, and that most species have a small number of FolB genes that diverged after separation of the lineages leading to families. The substantial divergence of FolB homologs in Arabidopsis and other plants suggests that some of them may act on substrates other than DHN.

  1. The size effect in metal cutting

    Indian Academy of Sciences (India)

    Milton C Shaw

    2003-10-01

    When metal is removed by machining there is substantial increase in the specific energy required with decrease in chip size. It is generally believed this is due to the fact that all metals contain defects (grain boundaries, missing and impurity atoms, etc.), and when the size of the material removed decreases, the probability of encountering a stress-reducing defect decreases. Since the shear stress and strain in metal cutting is unusually high, discontinuous microcracks usually form on the metal-cutting shear plane. If the material being cut is very brittle, or the compressive stress on the shear plane is relatively low, microcracks grow into gross cracks giving rise to discontinuous chip formation. When discontinuous microcracks form on the shear plane they weld and reform as strain proceeds, thus joining the transport of dislocations in accounting for the total slip of the shear plane. In the presence of a contaminant, such as CCl4 vapour at a low cutting speed, the rewelding of microcracks decreases, resulting in decrease in the cutting force required for chip formation. A number of special experiments are described in the paper that support the transport of microcracks across the shear plane, and the important role compressive stress plays on the shear plane. Relatively recently, an alternative explanation for the size effect in cutting was provided based on the premise that shear stress increases with increase in strain rate. When an attempt is made to apply this to metal cutting by Dinesh et al (2001) it is assumed in the analysis that the von Mises criterion pertains to the shear plane. This is inconsistent with the experimental findings of Merchant. Until this difficulty is taken care of, together with the promised experimental verification of the strain rate approach, it should be assumed that the strain rate effect may be responsible for some notion of the size effect in metal cutting. However, based on the many experiments discussed here, it is

  2. Effects of cutting and maturity on antioxidant activity of fresh-cut tomatoes

    NARCIS (Netherlands)

    Moreira Lana, M.; Tijskens, L.M.M.; Kooten, van O.

    2005-01-01

    To investigate the changes in total antioxidant activity of fresh-cut tomato during storage, tomato fruits harvested at three different stages of maturity were cut into 7-mm thick slices and stored at 5°C. Intact (control) fruits were stored in the same conditions. The antioxidant activity was evalu

  3. Effects of cutting and maturity on antioxidant activity of fresh-cut tomatoes

    NARCIS (Netherlands)

    Moreira Lana, M.; Tijskens, L.M.M.

    2006-01-01

    To investigate the changes in total antioxidant activity of fresh-cut tomato during storage, tomato fruits harvested at three different stages of maturity were cut into 7-mm thick slices and stored at 5 °C. Intact fruits were stored in the same conditions as a control. The antioxidant activity was e

  4. Effects of cutting and maturity on antioxidant activity of fresh-cut tomatoes

    NARCIS (Netherlands)

    Moreira Lana, M.; Tijskens, L.M.M.

    2006-01-01

    To investigate the changes in total antioxidant activity of fresh-cut tomato during storage, tomato fruits harvested at three different stages of maturity were cut into 7-mm thick slices and stored at 5 °C. Intact fruits were stored in the same conditions as a control. The antioxidant activity was e

  5. Laser cutting of rectangular geometry into aluminum alloy: Effect of cut sizes on thermal stress field

    Science.gov (United States)

    Akhtar, Sohail; Kardas, Omer Ozgur; Keles, Omer; Yilbas, Bekir Sami

    2014-10-01

    Laser cutting of a rectangular geometry into aluminum alloy 2024 is carried out. Temperature and stress fields are predicted in the cutting section using the ABAQUS finite element code in line with the experimental conditions. Effect of the size of the rectangular geometry on the thermal stress fields is examined in the cutting section. Temperature predictions are validated through the thermocouple data. To identify the morphological changes in the cutting section, an experiment is carried out and the resulting cutting sections are examined under optical and scanning electron microscopes. It is found that temperature and stress fields are affected by the size of the rectangular cut geometry. Temperature and von Mises stress attains higher values for small size rectangular geometry as compared to its counterpart corresponding to the large size geometry. Laser cut sections are free from large size asperities including sideways burning and out-off flatness at the cut edges. Locally scattered some small dross attachments are observed at the kerf exit.

  6. A major barley allergen associated with baker's asthma disease is a glycosylated monomeric inhibitor of insect alpha-amylase: cDNA cloning and chromosomal location of the gene.

    Science.gov (United States)

    Mena, M; Sanchez-Monge, R; Gomez, L; Salcedo, G; Carbonero, P

    1992-11-01

    A 14.5 kDa barley endosperm protein that is a major allergen in baker's asthma disease, as previously shown by both in vitro (IgE binding) and in vivo tests, has been identified as a glycosylated monomeric member of the multigene family of inhibitors of alpha-amylase/trypsin from cereals. A cDNA encoding this allergen (renamed BMAI-1) has been isolated and characterized. The deduced sequence for the mature protein, which is 132 residues long, is identical in its N-terminal end to the 20 amino acid partial sequence previously determined from the purified allergen, and fully confirms that it is a member of the multigene family of cereal inhibitors. Southern-blot analysis of wheat/barley addition lines using the insert in the BMAI-1 cDNA clone as a probe, has led to the location of the allergen gene (Iam1) in barley chromosome 2, while another related member of this protein family, the barley dimeric alpha-amylase inhibitor BDAI-1 gene (Iad1) has been located in chromosome 6. Iam1 is the first member of this inhibitor family in cereals to be assigned to chromosome group 2, thus extending the dispersion of genes in the family to five out of the seven homology groups of chromosomes in wheat and barley (chromosomes 2, 3, 4, 6 and 7).

  7. Two metallothionein gene family members in buckwheat: Expression analysis in flooding stress using Real Time RT-PCR technology

    Directory of Open Access Journals (Sweden)

    Majić Dragana B.

    2008-01-01

    Full Text Available Metallothioneins (MTs are an extensive and diverse family of small cysteine-rich proteins with metal-binding ability that are involved in metal homeostasis and detoxification. Two cDNA clones of the MT3 type, differing in 3’ UTRs, were isolated from the developing buckwheat seed cDNA library. Following sequence analyses, expression profiles during flooding stress were monitored by Real Time RT PCR technology.

  8. The universal cut function and type II metrics

    Energy Technology Data Exchange (ETDEWEB)

    Kozameh, Carlos [FaMaF, University of Cordoba, Cordoba (Argentina); Newman, E T [Department of Physics and Astronomy, University of Pittsburgh, Pittsburgh, PA 15260 (United States); Santiago-Santiago, J G [Facultad de Ciencias Fisico Matematicas de la Universidad Autonoma de Puebla, Apartado Postal 1152, 72001, Puebla, Pue. (Mexico); Silva-Ortigoza, Gilberto [Facultad de Ciencias Fisico Matematicas de la Universidad Autonoma de Puebla, Apartado Postal 1152, 72001, Puebla, Pue. (Mexico)

    2007-04-21

    In analogy with classical electromagnetic theory, where one determines the total charge and both electric and magnetic multipole moments of a source from certain surface integrals of the asymptotic (or far) fields, it has been known for many years-from the work of Hermann Bondi-that the energy and momentum of gravitational sources could be determined by similar integrals of the asymptotic Weyl tensor. Recently, we observed that there were certain overlooked structures, defined at future null infinity, that allowed one to determine (or define) further properties of both electromagnetic and gravitating sources. These structures, families of complex 'slices' or 'cuts' of Penrose's I{sup +}, are referred to as universal cut functions. In particular, one can define from these structures a (complex) centre of mass (and centre of charge) and its equations of motion-with rather surprising consequences. It appears as if these asymptotic structures contain, in their imaginary part, a well-defined total spin-angular momentum of the source. We apply these ideas to the type II algebraically special metrics, both twisting and twist free.

  9. Influence of cutting conditions on chip side curl

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo

    2004-01-01

    The paper describes the influence of local variations of contact length, cutting speed and material constraint, showing the effect of lubrication, on the side curl of the chip. The following examples are illustrated by experiments: cutting of a tube vs. cutting of a bar; cutting using a tool...

  10. Influence of cutting conditions on chip side curl

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo

    2004-01-01

    The paper describes the influence of local variations of contact length, cutting speed and material constraint, showing the effect of lubrication, on the side curl of the chip. The following examples are illustrated by experiments: cutting of a tube vs. cutting of a bar; cutting using a tool with...

  11. Investigations of Cutting Fluid Performance Using Different Machining Operations

    DEFF Research Database (Denmark)

    De Chiffre, Leonardo; Belluco, Walter

    2002-01-01

    An analysis of cutting fluid performance in dif-ferent metal cutting operations is presented based on performance criteria, work material and fluid type. Cutting fluid performance was evaluated in turning, drilling, reaming and tapping operations, with respect to tool life, cutting forces and prod...

  12. Evaluation of Cutting Fluids in Multiple Reaming of Stainless Steel

    DEFF Research Database (Denmark)

    Belluco, Walter; Zeng, Z.; De Chiffre, Leonardo

    2001-01-01

    An investigation on the effect of different cutting fluids in reaming is presented. The performance of three water based cutting fluids and one cutting oil was compared to that of a reference water based commercial product by measurement of cutting forces, surface roughness and part accuracy. Three...

  13. 7 CFR 58.413 - Cutting and packaging rooms.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Cutting and packaging rooms. 58.413 Section 58.413....413 Cutting and packaging rooms. When small packages of cheese are cut and wrapped, separate rooms shall be provided for the cleaning and preparation of the bulk cheese and for the cutting and...

  14. 21 CFR 882.4275 - Dowel cutting instrument.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dowel cutting instrument. 882.4275 Section 882...) MEDICAL DEVICES NEUROLOGICAL DEVICES Neurological Surgical Devices § 882.4275 Dowel cutting instrument. (a) Identification. A dowel cutting instrument is a device used to cut dowels of bone for bone grafting....

  15. STUDY ON SUB-DRY CUTTING GCr12

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Through the comparison study on cutting force, cutting temperature and machined surface quality with the sub-dry cutting traditional cooling method, it is shown that sub-dry cutting can retard the wear of the tooled parts. It is beneficial to realize the production without pollution and meet the demand of clean environment.

  16. Material testing of copper by extrusion-cutting

    DEFF Research Database (Denmark)

    Segalina, F.; De Chiffre, Leonardo

    2017-01-01

    was developed and implemented on a CNC lathe. An investigation was carried out extrusion-cutting copper discs using high-speed-steel cutting tools at 100 m/min cutting speed. Flow stress values for copper under machining-relevant conditions were obtained from measurement of the extrusion-cutting force...

  17. 49 CFR 236.52 - Relayed cut-section.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Relayed cut-section. 236.52 Section 236.52...: All Systems Track Circuits § 236.52 Relayed cut-section. Where relayed cut-section is used in... shall be open and the track circuit shunted when the track relay at such cut-section is in deenergized...

  18. Recent results in mirror based high power laser cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove; Nielsen, Jakob Skov; Elvang, Mads

    2004-01-01

    In this paper, recent results in high power laser cutting, obtained in reseach and development projects are presented. Two types of mirror based focussing systems for laser cutting have been developed and applied in laser cutting studies on CO2-lasers up to 12 kW. In shipyard environment cutting...

  19. Trim cut machining and surface integrity analysis of Nimonic 80A alloy using wire cut EDM

    Directory of Open Access Journals (Sweden)

    Amitesh Goswami

    2017-02-01

    Full Text Available This present work deals with the features of trim cut wire EDM machining of Nimonic 80A in terms of machining parameters, to predict material removal rate (MRR, surface roughness (Ra, wire wear ratio (WWR and microstructure analysis. Trim cut is performed after rough cut to remove the rough layer deposited after machining due to melting and re-solidification of the eroded metal from workpiece as well as from wire electrode. Taguchi’s design of experiments methodology has been used for planning and designing the experiments. The relative significance of various factors has also been evaluated and analyzed using ANOVA. The results clearly indicate trim cut potential for high surface finish compared to rough cut machining.

  20. The $\\mathcal{Q}$-cut Representation of One-loop Integrands and Unitarity Cut Method

    CERN Document Server

    Huang, Rijun; Rao, Junjie; Zhou, Kang; Feng, Bo

    2015-01-01

    Recently, a new construction for complete loop integrands of massless field theories has been proposed, with on-shell tree-level amplitudes delicately incorporated into its algorithm. This new approach reinterprets integrands in a novel form, namely the $\\mathcal{Q}$-cut representation. In this paper, by deriving one-loop integrands as examples, we elaborate in details the technique of this new representation, e.g., the summation over all possible $\\mathcal{Q}$-cuts as well as helicity states for the non-scalar internal particle in the loop. Moreover, we show that the integrand in the $\\mathcal{Q}$-cut representation naturally reduces to the integrand in the traditional unitarity cut method for each given cut channel, providing a cross-check for the new approach.

  1. Photonic mesophases from cut rod rotators

    Energy Technology Data Exchange (ETDEWEB)

    Stelson, Angela C.; Liddell Watson, Chekesha M., E-mail: cml66@cornell.edu [Materials Science and Engineering, Cornell University, Ithaca, New York 14853 (United States); Avendano, Carlos [Chemical Engineering and Analytical Science, The University of Manchester, Manchester M13 9PL (United Kingdom)

    2016-01-14

    The photonic band properties of random rotator mesophases are calculated using supercell methods applied to cut rods on a hexagonal lattice. Inspired by the thermodynamic mesophase for anisotropic building blocks, we vary the shape factor of cut fraction for the randomly oriented basis. We find large, stable bandgaps with high gap isotropy in the inverted and direct structures as a function of cut fraction, dielectric contrast, and filling fraction. Bandgap sizes up to 34.5% are maximized at high dielectric contrast for rods separated in a matrix. The bandgaps open at dielectric contrasts as low as 2.0 for the transverse magnetic polarization and 2.25 for the transverse electric polarization. Additionally, the type of scattering that promotes the bandgap is correlated with the effect of disorder on bandgap size. Slow light properties are investigated in waveguide geometry and slowdown factors up to 5 × 10{sup 4} are found.

  2. Manipulation and cutting of carbon nanotubes

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Nanomanipulation plays an important role in nanofabrication, it is also a technology necessary in exploring the secrets of nanoworld, and it thus beco mesa start point to research future nanomachine. In this study, manipulation and cutting of carbon nanotubes have been conducted in order to examine whether we can move a nanocomponent from one site to another by using the tip of atomic fo rce microscope (AFM). The technique may also be valuable for providing the const ructive materials of nanofabrication. While exploring the method for manipulatin g and cutting of nanotubes, some new phenomena have been observed during the process. Results show that carbon nanotubes present a feature of deformation combin ing bending and distortion when subjected to large mechanical forces exerted by the tip of AFM. In special cases, long carbon nanotubes can be cut into two part s, by which we can remove the part where crystal lattice is flawed, and therefor e a perfect nanocomponent can be obtained.

  3. ENVIRONMENTALLY REDUCING OF COOLANTS IN METAL CUTTING

    Directory of Open Access Journals (Sweden)

    Veijo KAUPPINEN

    2012-11-01

    Full Text Available Strained environment is a global problem. In metal industries the use of coolant has become more problematic in terms of both employee health and environmental pollution. It is said that the use of coolant forms approximately 8 - 16 % of the total production costs.The traditional methods that use coolants are now obviously becoming obsolete. Hence, it is clear that using a dry cutting system has great implications for resource preservation and waste reduction. For this purpose, a new cooling system is designed for dry cutting. This paper presents the new eco-friendly cooling innovation and the benefits gained by using this method. The new cooling system relies on a unit for ionising ejected air. In order to compare the performance of using this system, cutting experiments were carried out. A series of tests were performed on a horizontal turning machine and on a horizontal machining centre.

  4. COMPUTER AIDED DESIGN OF CUTTING TOOLS

    Directory of Open Access Journals (Sweden)

    Jakub Matuszak

    2015-11-01

    Full Text Available Correct and stable machining process requires an appropriate cutting tool. In most cases the tool can be selected by using special tool catalogs often available in online version. But in some cases there is a need to design unusual tools, for special treatment, which are not available in tool manufacturers’ catalogs. Proper tool design requires strength and geometric calculations. Moreover, in many cases specific technical documentation is required. By using Computer Aided Design of cutting tools this task can be carried out quickly and with high accuracy. Cutting tool visualization in CAD programs gives a clear overview of the design process. Besides, these programs provide the ability to simulate real machining process. Nowadays, 3D modeling in CAD programs is a fundamental tool for engineers. Therefore, it is important to use them in the education process.

  5. Basics of cutting and abrasive processes

    CERN Document Server

    Toenshoff, Hans Kurt

    2013-01-01

    Manufacturing is the basic industrial activity generating real value. Cutting and abrasive technologies are the backbone of precision production in machine, automotive and aircraft building as well as of production of consumer goods. We present the knowledge of modern manufacturing in these technologies on the basis of scientific research. The theory of cutting and abrasive processes and the knowledge about their application in industrial practice are a prerequisite for the studies of manufacturing science and an important part of the curriculum of the master study in German mechanical engineering. The basis of this book is our lecture “Basics of cutting and abrasive processes” (4 semester hours/3 credit hours) at the Leibniz University Hannover, which we offer to the diploma and master students specializing in manufacturing science.

  6. Laser Cutting of Materials of Various Thicknesses

    Directory of Open Access Journals (Sweden)

    Martin Grepl

    2012-01-01

    Full Text Available Thise paper deals with the application of laser technology and optimizing the parameters for cutting nickel alloy. The theoretical part of the paper describes various types of lasers, their principles and usage. The experimental part focuses on optimizing the section parameteres of Haynes 718 alloy using a CO2 gas laser. This alloy is employed in the production of components for the aircraft industry. The experiment was performed on the Wibro Delta laser system designed for sizable parts. The actual section is measured with respect to its quality and any accompanying side effects that occur during the process. In this case, laser output and cutting speed were the parameters with most influence on the final cut. The summary explains the results achieved in a metallographic laboratory.

  7. Cut Locus Construction using Deformable Simplicial Complexes

    DEFF Research Database (Denmark)

    Misztal, Marek Krzysztof; Bærentzen, Jakob Andreas; Anton, François;

    2011-01-01

    In this paper we present a method for appproximating cut loci for a given point p on Riemannian 2D manifolds, closely related to the notion of Voronoi diagrams. Our method finds the cut locus by advecting a front of points equally distant from p along the geodesics originating at p and finding...... the domain to have disk topology. We test our method for tori of revolution and compare our results to the benchmark ones from [2]. The method, however, is generic and can be easily adapted to construct cut loci for other manifolds of genera other than 1....... the lines of self-intersections of the front in the parametric space. This becomes possible by using the deformable simplicial complexes (DSC, [1]) method for deformable interface tracking. DSC provide a simple collision detection mechanism, allows for interface topology control, and does not require...

  8. Revised sequence and expression of cyclin B cDNA from the starfish Asterina pectinifera.

    Science.gov (United States)

    Miyake, Y; Deshimaru, S; Toraya, T

    2001-05-01

    Cyclin B cDNA was cloned from the ovary of the starfish Asterina pectinifera and analyzed by RT-PCR and 3'- and 5'-RACE techniques. The cDNA consists of a 0.13-kb upstream untranslated region, a 1.22-kb coding region, and a 0.86-kb downstream untranslated region. The open reading frame encoded a polypeptide of 404 amino acid residues with a calculated molecular weight of 45,692. All the characteristic sequences, such as destruction and cyclin boxes, cyclin B motif, and cytoplasmic retention and nuclear export signals, were found in the newly cloned cyclin B cDNA. The deduced amino acid sequence of the cyclin B cDNA was highly homologous in the middle and carboxy terminal regions to that from mature eggs of the same organism, but quite different in the amino terminal region. Evidence was obtained which suggested that this cyclin B is expressed in immature and maturing oocytes and is the same as that cloned from mature eggs.

  9. Strategies to enhance immunogenicity of cDNA vaccine encoded antigens by modulation of antigen processing

    NARCIS (Netherlands)

    Platteel, Anouk C M; Marit de Groot, A; Andersen, Peter; Ovaa, Huib; Kloetzel, Peter M; Mishto, Michele; Sijts, Alice J A M

    2016-01-01

    Most vaccines are based on protective humoral responses while for intracellular pathogens CD8(+) T cells are regularly needed to provide protection. However, poor processing efficiency of antigens is often a limiting factor in CD8(+) T cell priming, hampering vaccine efficacy. The multistage cDNA va

  10. Isolation of 24 novel cDNA fragments from microdis—sected human chromosome band

    Institute of Scientific and Technical Information of China (English)

    ZHANGMIN; LONGYU; 等

    1998-01-01

    The strategy of isolating the band0specific expression fragments from a probe pool generated by human chromosome microdissection was reported.A chromosome 14q 24.3 band-specific single copy DNA pool was constructed based on this probe pool.Using total DNA of the pool as probe to hybridize the human marrow cDNA library,68 primary positive clones were selected from 5×105 cDNA clones.Among these primary clones,32 secondary clones were obtained after second-round screening and designed as cFD14-1-32.Finally,24 band-specific expression fragments were identified from these 32 positive clones by DNA hybridization.Those band-specific clones can hybridize to both 14q24.3 DNA and human genomic DNA but cann't hybridize to 17q11-12 DNA,Partial sequences of 13 fragments of them were sequenced and idenfified as novel cDNA sequences,and these sequences were proved to have some homology with known genes in NCBI database.Analysis of expression spectrum of cFD 14-1 suggested that the cDNA fragments thus obtained should be used to isolate the genes can not been cloned in 14q24.3 region.

  11. Molecular cloning and expression of a novel human cDNA containing CAG repeats.

    Science.gov (United States)

    Takeuchi, T; Chen, B K; Qiu, Y; Sonobe, H; Ohtsuki, Y

    1997-12-19

    A novel human cDNA containing CAG repeats, designated B120, was cloned by PCR amplification. An approximately 300-bp 3' untranslated region in this cDNA was followed by a 3426-bp coding region containing the CAG repeats. A computer search failed to find any significant homology between this cDNA and previously reported genes. The number of CAG trinucleotide repeats appeared to vary from seven to 12 in analyses of genomic DNA from healthy volunteers. An approximately 8-kb band was detected in brain, skeletal muscle and thymus by Northern blot analysis. The deduced amino-acid sequence had a polyglutamine chain encoded by CAG repeats as well as glutamine- and tyrosine-rich repeats, which has also been reported for several RNA binding proteins. We immunized mice with recombinant gene product and established a monoclonal antibody to it. On Western immunoblotting, this antibody detected an approximately 120-kDa protein in human brain tissue. In addition, immunohistochemical staining showed that the cytoplasm of neural cells was stained with this antibody. These findings indicated that B120 is a novel cDNA with a CAG repeat length polymorphism and that its gene product is a cytoplasmic protein with a molecular mass of 120 kDa.

  12. cDNA, genomic cloning and sequence analysis of ribosomal protein ...

    African Journals Online (AJOL)

    enoh

    2012-03-13

    Mar 13, 2012 ... melanoleuca) using reverse transcriptase-polymerase chain reaction (RT-PCR) and touchdown-PCR ... RNA species and approximately 80 structurally distinct ... a reverse transcription kit with Oligo dT as the primers according to ... 2). On the basis of the high identity, we concluded that the cDNA isolated is ...

  13. cDNA - ASTRA | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available English ]; } else { document.getElementById(lang).innerHTML= '[ Japanese | English ]'; } } window.onload = ...ies Species name Chr. No. Chromosome No. Strand Strand Gene name Gene name UniGene ID UniGene ID cDNA UniGen

  14. GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN CDNA MICROARRAY ANALYSES

    Science.gov (United States)

    GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN cDNA MICROARRAY ANALYSESB.S. Pukazhenthi1, J. C. Rockett2, M. Ouyang3, D.J. Dix2, J.G. Howard1, P. Georgopoulos4, W.J. J. Welsh3 and D. E. Wildt11Department of Reproductiv...

  15. Observation of intermittency in gene expression on cDNA microarrays

    CERN Document Server

    Peterson, L E

    2002-01-01

    We used scaled factorial moments to search for intermittency in the log expression ratios (LERs) for thousands of genes spotted on cDNA microarrays (gene chips). Results indicate varying levels of intermittency in gene expression. The observation of intermittency in the data analyzed provides a complimentary handle on moderately expressed genes, generally not tackled by conventional techniques.

  16. CLONING AND CHARACTERIZATION OF CDNA ENCODING GIARDIA LAMBLIA d-GIARDIN

    Science.gov (United States)

    A cDNA coding for d-giardin was cloned from Giardia lamblia trophozoites in order to localize the protein and study its function in mediating surface attachment. Recombinant d-giardin antigen was produced in Escherichia coli as a poly-histidine fusion protein and was purified by affinity chromatogr...

  17. Isolation of carrot Argonaute1 from subtractive somatic embryogenesis cDNA library.

    Science.gov (United States)

    Takahata, Kiminori

    2008-03-01

    Carrot Argonaute1 (C-Ago1) was isolated from a subtractive cDNA library to obtain somatic embryogenesis related genes. C-Ago1 has three conserved domains, which are found in all other Argonautes. C-Ago1 has specific expression during somatic embryogenesis, which indicates that microRNA gene expression controlling system is required for somatic embryogenesis.

  18. Isolation of RNA and DNA from leukocytes and cDNA synthesis.

    NARCIS (Netherlands)

    Jansen, J.H.; Reijden, B.A. van der

    2006-01-01

    In this chapter, methods to isolate RNA and DNA from human leukocytes for the subsequent use in molecular diagnostic tests are described. In addition, protocols for cDNA synthesis are given, both for the use in conventional reverse transcription (RT)-polymerase chain reaction (PCR), and for the use

  19. Cloning and Sequence Analysis of cDNA Encoding MRJP3 of Apis cerana cerana

    Institute of Scientific and Technical Information of China (English)

    SU Song-kun; ZHNEG Huo-qing; CHEN Sheng-lu; ZHONG Bo-xiong; Stefan Albert

    2005-01-01

    By screening the worker (Apis cerana cerana) heads cDNA library using a fragment of the mrjp3 gene ofApis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 (AccMRJP3) cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly (A) tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame (ORF) with 1 779 bp encoding 593 amino acids. The un-translated regions (UTR) of the 5' end and 3' end are 46 bp and 160 bp in length,respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them.

  20. Significance of RNA reference in tumour-related gene expression analyses by cDNA array.

    Science.gov (United States)

    Laytragoon-Lewin, Nongnit; Lagerlund, Magnus; Lundgren, Jan; Nordlander, Britt; Elmberger, Göran; Södergren, Towe; Lagerros, Christofer; Rutqvist, Lars Erik; Lewin, Freddi

    2005-01-01

    The cDNA array technique is an efficient approach for studying the expression of a large number of genes in a single experiment. The cDNA array analysis indicates the relative level of corresponding gene expression from a specimen and a reference. Our investigation was performed to address the significance of reference RNA on the outcome of the cancer-related gene expression profile obtained from cDNA array analysis. Human head and neck squamous cell carcinoma (HNSCC) biopsies and 5 sources of RNA reference were used for this purpose. In these biopsies, each individual patient expressed a unique set of genes both in normal and tumour tissue. It is important to note that 5 striking patterns of tumour-related gene expression were obtained according to the 5 references used. Significant differences in 60%, 16%, 15% and 15% of the genes expressed were shown when autologous normal matched tissue biopsy references were compared to pooled cell lines, allogenic normal mixed cell types, tumours or allogenic normal matched cell type references, respectively. Thus, theoretically and our study suggested that patient autologous normal cells matching with the tumour type should be the most suitable reference in cDNA array for the identification of individual tumour gene profiles with clinical purpose.

  1. Cloning chromosome specific genes by reciprocal probing of arrayed cDNA and cosmid libraries

    Energy Technology Data Exchange (ETDEWEB)

    Yazdani, A.; Lee, C.C.; Wehnert, M. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1994-09-01

    A human gene map will greatly facilitate the association of genes to single locus diseases and provide candidates for genes involved in complex genetic traits. Given the estimated 100,000 human genes an integrated strategy with a high throughput approach for isolation and mapping of expressed sequences is needed to create such a gene map. We have developed an approach that allows high throughput gene isolation and mapping using arrayed genomic and cDNA lambda libraries. Reciprocal probing of the arrayed genomic and cDNA cosmic libraries can rapidly establish cDNA-cosmid associations. Fluorescence in situ hybridization (FISH) chromosomal mapping and expressed sequence tag/sequence tag site (EST/STS) primers generated from DNA sequence of PCR-based mapping using somatic hybrid cell line mapping panels were utilized to characterize further the hybridization-based cDNA cosmid association. We have applied this approach to chromosome 17 using a placental cDNA library and have identified a total of 30 genes out of which 11 are novel. Furthermore seven cDNAs were mapped to 17q21 in this study, providing novel candidate genes for BRCA-1 gene for early onset breast cancer. The results of our study clearly show that an integration of an expression map into physical and genetic maps can provide candidate genes for human diseases that have been mapped to specific regions. This approach combined with the current physical mapping efforts could efficiently provide a detailed human gene map.

  2. Construction of full-length cDNA library of white flower Salvia ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... base for further study on the structure and function of these cDNAs. Key words: white flower ..... precipitated by ethylene glycol monobutyl ether is high purity. The A260/A230 ... gives high feasibility for cloning full length cDNA.

  3. Complete amino acid sequence of human intestinal aminopeptidase N as deduced from cloned cDNA

    DEFF Research Database (Denmark)

    Cowell, G M; Kønigshøfer, E; Danielsen, E M

    1988-01-01

    The complete primary structure (967 amino acids) of an intestinal human aminopeptidase N (EC 3.4.11.2) was deduced from the sequence of a cDNA clone. Aminopeptidase N is anchored to the microvillar membrane via an uncleaved signal for membrane insertion. A domain constituting amino acid 250...

  4. Discovery and analysis of pancreatic adenocarcinoma genes using cDNA microarrays

    Institute of Scientific and Technical Information of China (English)

    Gang Jin; Xian-Gui Hu; Kang Ying; Yan Tang; Rui Liu; Yi-Jie Zhang; Zai-Ping Jing; Yi Xie; Yu-Min Mao

    2005-01-01

    AIM: To study the pathogenetic processes and the role of gene expression by microarray analyses in expediting our understanding of the molecular pathophysiology of pancreatic adenocarcinoma, and to identify the novel cancer-associated genes.METHODS: Nine histologically defined pancreatic head adenocarcinoma specimens associated with clinical data were studied. Total RNA and mRNA were isolated and labeled by reverse transcription reaction with Cy5 and Cy3 for cDNA probe. The cDNA microarrays that represent a set of 4 096 human genes were hybridized with labeled cDNA probe and screened for molecular profiling analyses.RESULTS: Using this methodology, 184 genes were screened out for differences in gene expression level after nine couples of hybridizations. Of the 184 genes,87 were upregulated and 97 downregulated, including 11 novel human genes. In pancreatic adenocarcinoma tissue, several invasion and metastasis related genes showed their high expression levels, suggesting that poor prognosis of pancreatic adenocarcinoma might have a solid molecular biological basis.CONCLUSION: The application of cDNA microarray technique for analysis of gene expression patterns is a powerful strategy to identify novel cancer-associated genes, and to rapidly explore their role in clinical pancreatic adenocarcinoma. Microarray profiles provide us new insights into the carcinogenesis and invasive process of pancreatic adenocarcinoma. Our results suggest that a highly organized and structured process of tumor invasion exists in the pancreas.

  5. Transcription analysis of apple fruit development using cDNA microarrays

    NARCIS (Netherlands)

    Soglio, V.; Costa, F.; Molthoff, J.W.; Weemen-Hendriks, M.; Schouten, H.J.; Gianfranceschi, L.

    2009-01-01

    The knowledge of the molecular mechanisms underlying fruit quality traits is fundamental to devise efficient marker-assisted selection strategies and to improve apple breeding. In this study, cDNA microarray technology was used to identify genes whose expression changes during fruit development and

  6. Isolation and cDNA cloning of somatolactin in rabbitfish (Siganus guttatus).

    Science.gov (United States)

    Ayson, F G; de Jesus, E G; Amemiya, Y; Moriyama, S; Hirano, T; Kawauchi, H

    1999-08-01

    We report the isolation and cDNA cloning of somatolactin (SL) from rabbitfish, Siganus guttatus. Rabbitfish SL was isolated from an alkaline extract of the pituitary glands by gel filtration chromatography on Sephadex G-100 and reversed-phase high-performance liquid chromatography. SL was monitored by immunoblotting with flounder SL antiserum. The preparation (yield: 0.86 mg/g wet tissues) contained two immunoreactive bands of 24 and 28 kDa on SDS-PAGE. Overlapping partial cDNA clones corresponding to teleost SLs were amplified by PCR from single-strand cDNA from pituitary glands. Excluding the poly(A) tail, rabbitfish SL cDNA is 1605 bp long. It contains a 693-bp open reading frame encoding a signal peptide of 24 amino acids (aa) and a mature protein of 207 aa. Rabbitfish SL has two possible N-glycosylation sites at positions 11 and 121 and seven half Cys residues. The deduced amino acid sequence shows over 80% identity with those of advanced teleosts like sea bream, red drum, and flounder, 76% with the salmonids, 57% with the eel, and 46% with the goldfish SL.

  7. Data of evolutionary structure change: 1CDNA-1CLMA [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1CDNA-1CLMA 1CDN 1CLM A A KSPEELKGIFEKYAAKE--GDPNQLSKEELKLLLQTEFP...--- LTE-EQIAEFKEAFALFDKDGDGTITTKELGTVMRSL-----GQNPTEAELQDMINEVDADGNGTIDFPEFLSLMARKMKEQDSEEELIEA...line> 1CLM A 1CLMA 1CLM A 1CLMA...fEVID> 2 1CLM A 1CLMA

  8. Rapid Amplification of cDNA Ends for RNA Transcript Sequencing in Staphylococcus.

    Science.gov (United States)

    Miller, Eric

    2016-01-01

    Rapid amplification of cDNA ends (RACE) is a technique that was developed to swiftly and efficiently amplify full-length RNA molecules in which the terminal ends have not been characterized. Current usage of this procedure has been more focused on sequencing and characterizing RNA 5' and 3' untranslated regions. Herein is described an adapted RACE protocol to amplify bacterial RNA transcripts.

  9. Vector sequences - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available [ Credits ] BLAST Search Image Search Home About Archive Update History Contact us ...od - Number of data entries 7 entries - Joomla SEF URLs by Artio About This Database Database Description Download License Update His...tory of This Database Site Policy | Contact Us Vector sequences - Budding yeast cDNA sequencing project | LSDB Archive ...

  10. GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN CDNA MICROARRAY ANALYSES

    Science.gov (United States)

    GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN cDNA MICROARRAY ANALYSESB.S. Pukazhenthi1, J. C. Rockett2, M. Ouyang3, D.J. Dix2, J.G. Howard1, P. Georgopoulos4, W.J. J. Welsh3 and D. E. Wildt11Department of Reproductiv...

  11. FIELD VALIDATION OF A SHEEPSHEAD MINNOW ESTROGEN-RESPONSIVE CDNA MACROARRAY

    Science.gov (United States)

    Hemmer, Michael J., Iris Knoebl, Becky L. Hemmer, Patrick Larkin, Peggy S. Harris and Nancy D. Denslow. In press. Field Validation of a Sheepshead Minnow Estrogen-Responsive cDNA Macroarray (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portl...

  12. Consistent errors in first strand cDNA due to random hexamer mispriming

    NARCIS (Netherlands)

    Van Gurp, T.P.; McIntyre, L.M.; Verhoeven, K.J.F.

    2013-01-01

    Priming of random hexamers in cDNA synthesis is known to show sequence bias, but in addition it has been suggested recently that mismatches in random hexamer priming could be a cause of mismatches between the original RNA fragment and observed sequence reads. To explore random hexamer mispriming as

  13. Characterization of mouse Clpp protease cDNA, gene, and protein

    DEFF Research Database (Denmark)

    Andresen, B S; Corydon, T J; Wilsbech, M

    2000-01-01

    characterized the mouse Clpp cDNA sequence, the organization of the mouse gene, the chromosomal localization, and the tissue-specific expression pattern. Moreover. the cellular localization and processing of mouse Clpp was studied by overexpression in transfected eukaryotic cells. Our results indicate...

  14. EXPRESSION PROFILING OF ESTROGENIC COMPOUNDS USING A SHEEPSHEAD MINNOW CDNA MACROARRAY

    Science.gov (United States)

    Larkin, Patrick, Leroy C. Folmar, Michael J. Hemmer, Arianna J. Poston and Nancy D. Denslow. 2003. Expression Profiling of Estrogenic Compounds Using a Sheepshead Minnow cDNA Macroarray. Environ. Health Perspect. 111(6):839-846. (ERL,GB 1171). A variety of anthropogenic c...

  15. Construction of full-length cDNA library of white flower Salvia ...

    African Journals Online (AJOL)

    African Journal of Biotechnology. Journal Home · ABOUT ... Double strand cDNA was cloned into pDNR-LIB vector. The number of clones, ... After BLAST analysis of some cDNAs, their possible functions were predicted. It is found that most of ...

  16. Detection of reverse transcriptase termination sites using cDNA ligation and massive parallel sequencing

    DEFF Research Database (Denmark)

    Kielpinski, Lukasz J; Boyd, Mette; Sandelin, Albin

    2013-01-01

    of these methods can be increased by applying massive parallel sequencing technologies.Here, we describe a versatile method for detection of reverse transcriptase termination sites based on ligation of an adapter to the 3' end of cDNA with bacteriophage TS2126 RNA ligase (CircLigase™). In the following PCR...

  17. Avoiding cross hybridization by choosing nonredundant targets on cDNA arrays

    DEFF Research Database (Denmark)

    Nielsen, Henrik Bjørn; Knudsen, Steen

    2002-01-01

    PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end....

  18. Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution.

    Science.gov (United States)

    Modahl, Cassandra M; Mackessy, Stephen P

    2016-06-01

    Envenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many venoms. Venomous snakes and their venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare venom complexity and protein sequences, venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in venoms, offering the possibility of obtaining cDNA sequences directly from venom. This study provides evidence that unknown full-length venom protein transcripts can be obtained from the venoms of multiple species from all major venomous snake families. These unknown venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each venom protein superfamily. This technique was used to assemble a partial venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan) by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within venom. Phospholipase A2 sequences were also recovered from the venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus), and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae) have stable mRNA present in their venoms. These cDNA sequences from venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing venoms that lack proteomic profiles and identify sequence characteristics indicating specific envenomation profiles. This approach, requiring only venom, provides

  19. Cloning and sequence analysis of a full-length cDNA of SmPP1cb encoding turbot protein phosphatase 1 beta catalytic subunit

    Science.gov (United States)

    Qi, Fei; Guo, Huarong; Wang, Jian

    2008-02-01

    Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase 1(PP1cb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPP1cb, by the rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPP1cb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5' untranslated region and 462 bp 3' untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B). And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPP1cb is extremely conserved in both amino acid and nucleotide acid levels compared with the PP1cb of other vertebrates and invertebrates, and its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXX ATGG, which is different from mammalian in two positions A-6 and G-3, indicating the possibility of different initiation of translation in turbot, and also the 3'UTR of SmPP1cb is highly diverse in the sequence similarity and length compared with other animals, especially zebrafish. The cloning and sequencing of SmPP1cb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.

  20. Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution.

    Directory of Open Access Journals (Sweden)

    Cassandra M Modahl

    2016-06-01

    Full Text Available Envenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many venoms. Venomous snakes and their venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare venom complexity and protein sequences, venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in venoms, offering the possibility of obtaining cDNA sequences directly from venom. This study provides evidence that unknown full-length venom protein transcripts can be obtained from the venoms of multiple species from all major venomous snake families. These unknown venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each venom protein superfamily. This technique was used to assemble a partial venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within venom. Phospholipase A2 sequences were also recovered from the venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus, and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae have stable mRNA present in their venoms. These cDNA sequences from venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing venoms that lack proteomic profiles and identify sequence characteristics indicating specific envenomation profiles. This approach, requiring only

  1. Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution

    Science.gov (United States)

    Modahl, Cassandra M.; Mackessy, Stephen P.

    2016-01-01

    Envenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many venoms. Venomous snakes and their venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare venom complexity and protein sequences, venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in venoms, offering the possibility of obtaining cDNA sequences directly from venom. This study provides evidence that unknown full-length venom protein transcripts can be obtained from the venoms of multiple species from all major venomous snake families. These unknown venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each venom protein superfamily. This technique was used to assemble a partial venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan) by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within venom. Phospholipase A2 sequences were also recovered from the venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus), and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae) have stable mRNA present in their venoms. These cDNA sequences from venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing venoms that lack proteomic profiles and identify sequence characteristics indicating specific envenomation profiles. This approach, requiring only venom, provides

  2. A framework for cut-over management

    Directory of Open Access Journals (Sweden)

    Guido Nageldinger

    2015-11-01

    Full Text Available The purpose of this paper is to provide a governance structure for IT-related projects in order to assure a safeguarded and timely transition to a productive environment. This transitioning, which rarely exceeds a weekend, is colloquially called ‘cut-over’, ‘rollout’ or ‘deployment’. The governance structure is defined in accordance with a set of project-specific deliverables for a cascade-type procedural project-management model, which is integrated within an Information Technology Infrastructure Library (ITIL-orientated service organization. This integration is illustrated by the use of a semi-agile release model. Due to the release model selected, which is particularly characterized by its bundling of projects for a release-specific rollout (as it is referred to in the project documentation, a new definition and interpretation of deployment from a generic ITIL perspective is required. The facilitated release model requires a distinction between a project-specific cut-over and a release-specific rollout. This separation gives rise to two types of go-live scenarios: one for each participating project and one for each release. Additionally, an interplay between cut-over planning for a project and rollout planning for a release becomes apparent. Projects should already incorporate cut-over related deliverables in the initial planning phase. Even though consulting methodologies such as ASAP (Accelerated SAP, recommend scattered, project-specific deliverables useful for cut-over planning, this publication offers an integrated approach on how to prepare systematically for a project-specific cut-over with all required deliverables. The framework provided maps out ITIL’s release and deployment process by means of IT projects; furthermore it allows IT projects to interface easily with the ITIL change-management process.

  3. ITER lip seal welding and cutting developments

    Energy Technology Data Exchange (ETDEWEB)

    Levesy, B.; Cordier, J.J.; Jokinen, T. [ITER Organization, Route de Vinon sur Verdon, 13115 Saint Paul Lez Durance (France); Kujanpää, V.; Karhu, M. [VTT Technical Research Centre of Finland (Finland); Le Barbier, R. [ITER Organization, Route de Vinon sur Verdon, 13115 Saint Paul Lez Durance (France); Määttä, T. [VTT Technical Research Centre of Finland (Finland); Martins, J.P.; Utin, Y. [ITER Organization, Route de Vinon sur Verdon, 13115 Saint Paul Lez Durance (France)

    2015-10-15

    Highlights: • Different TIG and Laser welding techniques are tested. • Twin spot laser welding techniques is the best. • Limited heat input gives a stable weld pool in all positions. • Penetrations is achieved. • Lip seal welding and cutting with a robotic arm is successfully performed on a representative mock-up. - Abstract: The welded lip seals form part of the torus primary vacuum boundary in between the port plugs and the vacuum vessel, and are classified as Protection Important Component. In order to refurbish the port plugs or the in-vessel components, port plugs have to be removed from the machine. The lip seal design must enable up to ten opening of the vacuum vessel during the life time operation of the ITER machine. Therefore proven, remote reliable cutting and re-welding are essential, as these operations need to be performed in the port cells in a nuclear environment, where human presence will be restricted. Moreover, the combination of size of the components to be welded (∼10 m long vacuum compatible thin welds) and the congested environment close to the core of the machine constraint the type and size of tools to be used. This paper describes the lip seal cutting and welding development programme performed at the VTT Technical Research Centre, Finland. Potential cutting and welding techniques are analyzed and compared. The development of the cutting, TIG and laser welding techniques on samples are presented. Effects of lip seal misalignments and optimization of the 2 welding processes are discussed. Finally, the manufacturing and test of the two 1.2 m × 1 m representative mock-ups are presented. The set-up and use of a robotic arm for the mock-up cutting and welding operations are also described.

  4. The perception of 3D shape from planar cut contours.

    Science.gov (United States)

    Egan, Eric J L; Todd, James T; Phillips, Flip

    2011-10-01

    A new computational analysis is described for estimating 3D shapes from orthographic images of surfaces that are textured with planar cut contours. For any given contour pattern, this model provides a family of possible interpretations that are all related by affine scaling and shearing transformations in depth, depending on the specific values of its free parameters that are used to compute the shape estimate. Two psychophysical experiments were performed in an effort to compare the model predictions with observers' judgments of 3D shape for developable and non-developable surfaces. The results reveal that observers' perceptions can be systematically distorted by affine scaling and shearing transformations in depth and that the magnitude and direction of these distortions vary systematically with the 3D orientations of the contour planes.

  5. Laser cutting of thick sheet metals: Residual stress analysis

    Science.gov (United States)

    Arif, A. F. M.; Yilbas, B. S.; Aleem, B. J. Abdul

    2009-04-01

    Laser cutting of tailored blanks from a thick mild steel sheet is considered. Temperature and stress field in the cutting sections are modeled using the finite element method. The residual stress developed in the cutting section is determined using the X-ray diffraction (XRD) technique and is compared with the predictions. The structural and morphological changes in the cut section are examined using the optical microscopy and scanning electron microscopy (SEM). It is found that temperature and von Mises stress increase sharply in the cutting section, particularly in the direction normal to the cutting direction. The residual stress remains high in the region close to the cutting section.

  6. Nonabelian cut diagrams and their applications

    CERN Document Server

    Lam, C S

    1996-01-01

    A new kind of cut diagram is introduced to sum Feynman diagrams with nonabelian vertices. Unlike the Cutkosky diagrams which compute the discontinuity of single Feynman diagrams, the nonabelian cut diagrams represent a resummation of both the real and the imaginary parts of Feynman diagrams related by permutations. Several applications of the technique are reported, including a resolution of the apparent inconsistency of the baryon problem in large-N_c QCD, a simplified calculation of high-energy low-order QCD diagrams, and progress made with this technique on the unitarization of the BFKL equation.

  7. On Non-Abelian Symplectic Cutting

    DEFF Research Database (Denmark)

    Martens, Johan; Thaddeus, Michael

    2012-01-01

    We discuss symplectic cutting for Hamiltonian actions of non-Abelian compact groups. By using a degeneration based on the Vinberg monoid we give, in good cases, a global quotient description of a surgery construction introduced by Woodward and Meinrenken, and show it can be interpreted in algebro......-geometric terms. A key ingredient is the `universal cut' of the cotangent bundle of the group itself, which is identified with a moduli space of framed bundles on chains of projective lines recently introduced by the authors....

  8. Cut Locus Construction using Deformable Simplicial Complexes

    DEFF Research Database (Denmark)

    Misztal, Marek Krzysztof; Bærentzen, Jakob Andreas; Anton, François

    2011-01-01

    In this paper we present a method for appproximating cut loci for a given point p on Riemannian 2D manifolds, closely related to the notion of Voronoi diagrams. Our method finds the cut locus by advecting a front of points equally distant from p along the geodesics originating at p and finding...... the lines of self-intersections of the front in the parametric space. This becomes possible by using the deformable simplicial complexes (DSC, [1]) method for deformable interface tracking. DSC provide a simple collision detection mechanism, allows for interface topology control, and does not require...

  9. Concrete Cutting Refinement for Crater Repair

    Science.gov (United States)

    2015-08-01

    ER D C/ G SL T R- 15 -2 9 Concrete Cutting Refinement for Crater Repair G eo te ch ni ca l a nd S tr uc tu re s La bo ra to ry...by ERDC, visit the ERDC online library at http://acwc.sdp.sirsi.net/client/default. ERDC/GSL TR-15-29 August 2015 Concrete Cutting Refinement...equipment in both soft and hard aggregate-mixed concrete . The investigated equipment included the Vermeer RW1236W and CC1531, Caterpillar SW45 and

  10. Cutting balloon angioplasty for intrastent restenosis treatment

    Directory of Open Access Journals (Sweden)

    João Orávio de Freitas Jr

    1999-05-01

    Full Text Available We describe here two patients with angiographic diagnosis of intrastent restenosis and regional myocardial ischemia. One stent restenosis was located in a native coronary artery and the other in a vein graft. Both were treated with cutting balloon angioplasty (CBA, inflated at low pressures. Angiographic success was obtained and both patients were discharged in the day after the procedure. Cutting balloon angioplasty using low inflation pressures achieved important luminal gains, in these two cases of intrastent restenosis. Further studies are necessary before the effectiveness of this procedure can be precisely defined.

  11. Expression and characterization of a cutinase (AnCUT2 from Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Ahmed Al-Tammar Khadijah

    2016-01-01

    Full Text Available Cutin hydrolase (EC 3.1.1.74, an extracellular polyesterase found in pollens, bacteria and fungi, is an efficient catalyst that exhibits hydrolytic activity on a variety of water-soluble esters, synthetic fibers, plastics and triglycerides. Thus, cutinase can be used in various applications such as ester synthesis, bio-scouring, food and detergent industries. Ancut2 is one of five genes encoding cutinases present in the Aspergillus niger ATCC 10574 genome. The cDNA of Ancut2 comprising of an open reading frame of 816 bp encoding a protein of 271 amino acid residues, was isolated and expressed in Pichia pastoris. The partially purified recombinant cutinase exhibited a molecular mass of approximately 40 kDa. The enzyme showed highest activity at 40°C with a preference for acidic pH (5.0-6.0. AnCUT2 showed hydrolytic activity towards various p-nitrophenyl esters with preference towards shorter chain esters such as p-nitrophenyl butyrate (C4. Scanning Electron Microscopy demonstrated that AnCUT2 was capable of modifying surfaces of synthetic polycaprolactone and polyethylene terephthalate plastics. The properties of this enzyme suggest that it may be applied in synthetic fiber modification and fruit processing industries.

  12. Laser capture microdissection: Arcturus(XT) infrared capture and UV cutting methods.

    Science.gov (United States)

    Gallagher, Rosa I; Blakely, Steven R; Liotta, Lance A; Espina, Virginia

    2012-01-01

    Laser capture microdissection (LCM) is a technique that allows the precise procurement of enriched cell populations from a heterogeneous tissue under direct microscopic visualization. LCM can be used to harvest the cells of interest directly or can be used to isolate specific cells by ablating the unwanted cells, resulting in histologically enriched cell populations. The fundamental components of laser microdissection technology are (a) visualization of the cells of interest via microscopy, (b) transfer of laser energy to a thermolabile polymer with either the formation of a polymer-cell composite (capture method) or transfer of laser energy via an ultraviolet laser to photovolatize a region of tissue (cutting method), and (c) removal of cells of interest from the heterogeneous tissue section. Laser energy supplied by LCM instruments can be infrared (810 nm) or ultraviolet (355 nm). Infrared lasers melt thermolabile polymers for cell capture, whereas ultraviolet lasers ablate cells for either removal of unwanted cells or excision of a defined area of cells. LCM technology is applicable to an array of applications including mass spectrometry, DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, proteomics discovery, and signal kinase pathway profiling. This chapter describes the unique features of the Arcturus(XT) laser capture microdissection instrument, which incorporates both infrared capture and ultraviolet cutting technology in one instrument, using a proteomic downstream assay as a model.

  13. A NEW METHOD TO CONSTRUCT A FULL-LENGTH cDNA LIBRARY OF HUMAN NORMAL BLADDER TISSUE

    Institute of Scientific and Technical Information of China (English)

    成瑜; 李旭; 陈葳; 杨玉琮; 赵乐

    2003-01-01

    Objective Using template-switch mechanism at the 5'-end of mRNA technique (SMART) to construct a full-length cDNA library of human normal bladder tissue. Methods The novel procedures used the template-switching activity of powerscript reverse transcriptase to synthesize and anchor first-strand cDNA in one step. Following reverse transcription, 5 cycles of PCR were performed using a modified oligo(dT) primer and an anchor primer to enrich the full-length cDNA population with 1.0 g human normal bladder poly(A)+RNA, then double-strand cDNA was synthesized. After digestion with sfiI and size-fractionation by CHROMA SPIN-400 columns, double-strand cDNA was ligated into λTripIEx2 vector and was packaged. We determined the titer of the primary library and the percentage of recombinant clones and finally amplified the library. Results The titer of the cDNA library constructed was 2.1×106 pfu*mL-1, and the amplified cDNA library was 6×1011 pfu*mL-1, the percentage of recombination clones was 99%. Conclusion Using SMART technique helps us to construct full-length cDNA library with high efficiency and high capacity which lays solid foundation for screening target genes of bladder diseases with probes and antibodies.

  14. Molecular cloning and expression analysis of cDNA ends of chicken neuropathy target esterase.

    Science.gov (United States)

    Chang, Ping-An; Sun, Quan; Ni, Xiao-Min; Qv, Feng-Qiong; Wu, Yi-Jun; Song, Fang-Zhou

    2008-03-10

    Neuropathy target esterase (NTE) was proposed as the initial target during the process of organophosphate-induced delayed neuropathy (OPIDN) in human and some sensitive animals. Adult hens are usually the animal model for experimental studies of OPIDN. However, little is known about the sequence and characteristics of chicken NTE. We report here the cloning of the 5' and 3' cDNA ends of chicken NTE through rapid amplification of cDNA ends (RACE) and their expression profiles in different tissues with northern blotting. The cloned 3' cDNA end of chicken NTE is 801 base pair (bp) in length with an open reading frame (ORF) of 379 bp. It contains a termination codon (TAG) and a 422-nucleotide noncoding sequence with the polyA sequence (GenBank accession no. DQ126678). The chicken NTE 5' cDNA end is 665 bp in length with an ORF of 552 bp. It contains an initiation codon (ATG) and a 113-bp untranslated region (GenBank accession no. DQ126677). The deduced proteins from 5' and 3' cDNA ends have a high degree of homology to humans and mouse NTE at the amino acid level. Chicken NTE is suggested to be a transmembrane protein by the transmembrane helix prediction of the deduced N-terminal sequence. The chicken NTE gene is expressed as a 4.5k b transcript in different tissues, including brain, kidney, liver and testis. Moreover, the mRNA expression of chicken NTE is highest in brain, and the mRNA levels of chicken NTE in testis, kidney and liver are about 75%, 47% and 24% of that in brain, respectively. These results should be helpful in cloning chicken full-length NTE gene.

  15. KLONING cDNA HORMON PERTUMBUHAN DARI IKAN GURAME (Osphronemus gouramy

    Directory of Open Access Journals (Sweden)

    Estu Nugroho

    2016-11-01

    Full Text Available Penelitian mengenai kloning cDNA pengkode hormon pertumbuhan ikan gurame telah dilakukan. Tujuan dari penelitian ini adalah untuk memperoleh sekuens DNA komplemen hormon pertumbuhan sebagai langkah awal dalam rangka pengembangan teknologi rekayasa genetik ikan gurame. Empat buah kelenjar hifopisa ikan gurame digunakan sebagai bahan bakunya dan dilakukan proses ekstraksi RNA total dari kelenjar hipofisa, dilanjutkan dengan sintesis cDNA, amplifikasi PCR, purifikasi fragmen DNA dari gel, ligasi produk PCR dengan vektor kloning, transformasi dan inkubasi bakteri, seleksi koloni bakteri putih, isolasi plasmid, dan sekuensing. Hasil sekuensing menunjukkan bahwa panjang produk amplifikasi PCR adalah 843 bp yang menyandikan 204 asam amino residu dan mengandung sekuens-sekuens yang konserf untuk gen hormon pertumbuhan (GH. Analisis homologi menunjukkan kesamaan sekuens hasil isolasi antara 52,4%--97,6% dengan gen GH ikan lainnya, dengan persentase homologi tertinggi adalah dengan ikan sepat. Dengan demikian dapat disimpulkan bahwa sekuens hasil isolasi merupakan sekuens gen GH. Dari hasil analisis sekuens terlihat bahwa gen GH ikan gurame secara evolusi adalah konserf. Research on cDNA cloning encoded the gouramy growth hormone was conducted. The aim of the research was to get complementary DNA, cDNA, sequences of growth hormone as an initial step to develop genetic engineering of gouramy fish. Four pituitary glands of the gouramy were taken and then processed with total RNA extraction, and continued with cDNA synthesis, PCR amplification, DNA fragment purification from the gel, PCR product legation with cloning vector, transformation and incubation of bacteria, white colony bacteria selection, plasmid isolation and sequencing analysis. Sequencing result showed that the amplified PCR product length had 834 bp, encoding 204 amino acid residue and contained conserve sequence for GH (growth hormone gen. Homolog analysis showed sequence similarity of

  16. Cloning and expression of a cDNA encoding human sterol carrier protein 2

    Energy Technology Data Exchange (ETDEWEB)

    Yamamoto, Ritsu; Kallen, C.B.; Babalola, G.O.; Rennert, H.; Strauss, J.F. III (Univ. of Pennsylvania School of Medicine, Philadelphia (United States)); Billheimer, J.T. (E.I. DuPont de Nemours, Inc., Wilmington, DE (United States))

    1991-01-15

    The authors report the cloning and expression of a cDNA encoding human sterol carrier protein 2 (SCP{sub 2}). The 1.3-kilobase (kb) cDNA contains an open reading frame which encompasses a 143-amino acid sequence which is 89% identical to the rat SCP{sub 2} amino acid sequence. The deduced amino acid sequence of the polypeptide reveals a 20-residue amino-terminal leader sequence in front of the mature polypeptide, which contains a carboxyl-terminal tripeptide (Ala-Lys-Leu) related to the peroxisome targeting sequence. The expressed cDNA in COS-7 cells yields a 15.3-kDa polypeptide and increased amounts of a 13.2-kDa polypeptide, both reacting with a specific rabbit antiserum to rat liver SCP{sub 2}. The cDNA insert hybridizes with 3.2- and 1.8-kb mRNA species in human liver poly(A){sup +} RNA. In human fibroblasts and placenta the 1.8-kb mRNA was most abundant. Southern blot analysis suggests either that there are multiple copies of the SCP{sub 2} gene in the human genome or that the SCP{sub 2} gene is very large. Coexpression of the SCP{sub 2} cDNA with expression vectors for cholesterol side-chain cleavage enzyme and adrenodoxin resulted in a 2.5-fold enhancement of progestin synthesis over that obtained with expression of the steroidogenic enzyme system alone. These findings are concordant with the notion that SCP{sub 2} plays a role in regulating steroidogenesis, among other possible functions.

  17. Cloning, expression and mapping of the full-length cDNA of human CCTβ subunit

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Chaperonins assist the proper folding of target proteins without being a part of the substrates. The eukaryotic cytosolic chaperonin, CCT-Chaperonin Containing TCP-1 (tailless complex polypeptide-1), is mainly involved in the formation of cytoskeletal proteins and is essential for cell viability. Mammalian CCT is commonly a protein complex composed of 7-9 subunit species. We have isolated a novel full-length cDNA from human testis cDNA library. This cDNA of 1935 bp contains a 1605 bp open reading frame (ORF) encoding 535 amino acids (aa). The deduced protein of the cDNA is highly homologous to the CCTβ subunit of saccharomyces cerevisiae, schizosaccharomyces pombe, caenorhabditis elegans and mouse, etc. Especially high homology (97%) is found between the deduced protein and mouse CCTb. On the basis of such high homology, the protein encoded by the new gene was proposed to be a human CCTβ subunit. Northern hybridization showed that human CCTβ gene is expressed as a transcript of about 2.0 kb in various tissues. Overexpression was seen in testis with the expression level 3-24 times of those in other tissues. The CCTβ gene was mapped to human chromosome 12q14 by Radiation Hybrid Mapping. Through homologous search, the 5′-end of the cDNA sequence was found to share intermittent regional homology with the 3′-end of human genomic sequence (U91327). The genomic structure of the 5′-end of CCTβ was also described in detail through comparative analysis.

  18. Recent results in mirror based high power laser cutting

    DEFF Research Database (Denmark)

    Olsen, Flemming Ove; Nielsen, Jakob Skov; Elvang, Mads

    2004-01-01

    In this paper, recent results in high power laser cutting, obtained in reseach and development projects are presented. Two types of mirror based focussing systems for laser cutting have been developed and applied in laser cutting studies on CO2-lasers up to 12 kW. In shipyard environment cutting...... speed increase relative to state-of-the-art cutting of over 100 % has been achieved....

  19. Analysis of beta-carotene hydroxylase gene cDNA isolated from the American oil-palm (Elaeis oleifera) mesocarp tissue cDNA library.

    Science.gov (United States)

    Bhore, Subhash J; Kassim, Amelia; Loh, Chye Ying; Shah, Farida H

    2010-09-20

    It is well known that the nutritional quality of the American oil-palm (Elaeis oleifera) mesocarp oil is superior to that of African oil-palm (Elaeis guineensis Jacq. Tenera) mesocarp oil. Therefore, it is of important to identify the genetic features for its superior value. This could be achieved through the genome sequencing of the oil-palm. However, the genome sequence is not available in the public domain due to commercial secrecy. Hence, we constructed a cDNA library and generated expressed sequence tags (3,205) from the mesocarp tissue of the American oil-palm. We continued to annotate each of these cDNAs after submitting to GenBank/DDBJ/EMBL. A rough analysis turned our attention to the beta-carotene hydroxylase (Chyb) enzyme encoding cDNA. Then, we completed the full sequencing of cDNA clone for its both strands using M13 forward and reverse primers. The full nucleotide and protein sequence was further analyzed and annotated using various Bioinformatics tools. The analysis results showed the presence of fatty acid hydroxylase superfamily domain in the protein sequence. The multiple sequence alignment of selected Chyb amino acid sequences from other plant species and algal members with E. oleifera Chyb using ClustalW and its phylogenetic analysis suggest that Chyb from monocotyledonous plant species, Lilium hubrid, Crocus sativus and Zea mays are the most evolutionary related with E. oleifera Chyb. This study reports the annotation of E. oleifera Chyb. ESTs - expressed sequence tags, EoChyb - Elaeis oleifera beta-carotene hydroxylase, MC - main cluster.

  20. Familial gigantism

    NARCIS (Netherlands)

    W.W. de Herder (Wouter)

    2012-01-01

    textabstractFamilial GH-secreting tumors are seen in association with three separate hereditary clinical syndromes: multiple endocrine neoplasia type 1, Carney complex, and familial isolated pituitary adenomas.

  1. Familial gigantism

    Directory of Open Access Journals (Sweden)

    Wouter W. de Herder

    2012-01-01

    Full Text Available Familial GH-secreting tumors are seen in association with three separate hereditary clinical syndromes: multiple endocrine neoplasia type 1, Carney complex, and familial isolated pituitary adenomas.

  2. Construction and Screening of an Expression cDNA Library from the Triactinomyxon Spores of Myxobolus cerebralis, the causative agent of Salmonid Whirling Diseases

    OpenAIRE

    Soliman, Hatem Mohamed Touhan

    2005-01-01

    The ZAP Express cDNA library was constructed using mRNA extracted from the triactinomyxon spores. First-strand cDNA was synthesized using Moloney Murine leukaemia virus reverse transcriptase. Following second-strand cDNA synthesis, the double-stranded cDNA was digested with Xho I restriction enzyme, cDNA fragments less than 400bp were removed and the remaining cDNA was ligated with the lambda ZAP Express vector. The recombinants were packaged in vitro using Gigapack III gold packaging extract...

  3. Purification, amino acid sequence, and cDNA cloning of trypsin inhibitors from onion (Allium cepa L.) bulbs.

    Science.gov (United States)

    Deshimaru, Masanobu; Watanabe, Akira; Suematsu, Keiko; Hatano, Maki; Terada, Shigeyuki

    2003-08-01

    Three protease inhibitors (OTI-1-3) have been purified from onion (Allium cepa L.) bulbs. Molecular masses of these inhibitors were found to be 7,370.2, 7,472.2, and 7,642.6 Da by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. Based on amino acid composition and N-terminal sequence, OTI-1 and -2 are the N-terminal truncated proteins of OTI-3. All the inhibitors are stable to heat and extreme pH. OTI-3 inhibited trypsin, chymotrypsin, and plasmin with dissociation constants of 1.3 x 10(-9) M, 2.3 x 10(-7) M, and 3.1 x 10(-7) M, respectively. The complete amino acid sequence of OTI-3 showed a significant homology to Bowman-Birk family inhibitors, and the first reactive site (P1) was found to be Arg17 by limited proteolysis by trypsin. The second reactive site (P1) was estimated to be Leu46, that may inhibit chymotrypsin. OTI-3 lacks an S-S bond near the second reactive site, resulting in a low affinity for the enzyme. The sequence of OTI-3 was also ascertained by the nucleotide sequence of a cDNA clone encoding a 101-residue precursor of the onion inhibitor.

  4. Cathepsin B from the white shrimp Litopenaeus vannamei: cDNA sequence analysis, tissues-specific expression and biological activity.

    Science.gov (United States)

    Stephens, A; Rojo, L; Araujo-Bernal, S; Garcia-Carreño, F; Muhlia-Almazan, A

    2012-01-01

    Cathepsin B is a cystein proteinase scarcely studied in crustaceans. Its function has not been clearly described in shrimp species belonging to the sub-order Dendrobranchiata, which includes the white shrimp Litopenaeus vannamei and other species from the Penaeidae family. Studies on vertebrates suggest that these lysosomal enzymes intracellularly hydrolize protein, as other cystein proteinases. However, the expression of the gene encoding the shrimp cathepsin B in the midgut gland was affected by starvation in a similar way as other digestive proteinases which extracellularly hydrolyze food protein. In this study the white shrimp L. vannamei cathepsin B (LvCathB) cDNA was sequenced, and characterized. Its gene expression was evaluated in various shrimp tissues, and changes in the mRNA amounts were compared with those observed on other digestive proteinases from the midgut gland during starvation. By using qRT-PCR it was found that LvCathB is expressed in most shrimp tissues except in pleopods and eye stalk. Changes on LvCathB mRNA during starvation suggest that the enzyme participates during intracellular protein hydrolysis but also, after food ingestion, it participates in hydrolyzing food proteins extracellularly as confirmed by the high activity levels we found in the gastric juice and midgut gland of the white shrimp.

  5. cDNA microarray analysis of disk abalone genes in gills and hemocytes after viral hemorrhagic septicemia virus (VHSV) challenge.

    Science.gov (United States)

    De Zoysa, Mahanama; Nikapitiya, Chamilani; Oh, Chulhong; Whang, Ilson; Shin, Hyun-Jin; Lee, Jehee

    2012-06-01

    A disk abalone Haliotis discus discus 4.2 K cDNA microarray was designed by selecting abalone expressed sequence tags (ESTs). Transcriptional profiles in gills and hemocytes were analyzed upon abalone challenged with viral hemorrhagic septicemia virus (VHSV) in order to select candidates for screening of immune response genes. Among the 4188 genes analyzed, 280 (6.6%) transcripts were changed their expression level in gills and hemocytes against VHSV challenge compared to control animals. Total of 88 and 65 genes were up-regulated in gills and hemocytes, respectively. These genes can be grouped under various immune-functional categories such as transcription factors (Krüppell-like factor; ETS-family transcription factor), inflammatory and apoptosis related genes (TNF superfamily members, Fas ligand), IFN regulatory proteins (IFN-44 like, interferon gamma-inducible lysosomal thiol reductase) and detoxification proteins (glutathione peroxidase). In contrast, 25 and 102 genes were shown down-regulation in gills and hemocytes, respectively. Among the differentially expressed transcripts, considerably higher numbers of ESTs were represented as either hypothetical (unknown) proteins or no GenBank match suggesting those may be novel genes associated with internal defense of abalone.

  6. Molecular Cloning and Characterization of Full-Length cDNA of Calmodulin Gene from Pacific Oyster Crassostrea gigas

    Directory of Open Access Journals (Sweden)

    Xing-Xia Li

    2016-01-01

    Full Text Available The shell of the pearl oyster (Pinctada fucata mainly comprises aragonite whereas that of the Pacific oyster (Crassostrea gigas is mainly calcite, thereby suggesting the different mechanisms of shell formation between above two mollusks. Calmodulin (CaM is an important gene for regulating the uptake, transport, and secretion of calcium during the process of shell formation in pearl oyster. It is interesting to characterize the CaM in oysters, which could facilitate the understanding of the different shell formation mechanisms among mollusks. We cloned the full-length cDNA of Pacific oyster CaM (cgCaM and found that the cgCaM ORF encoded a peptide of 113 amino acids containing three EF-hand calcium-binding domains, its expression level was highest in the mantle, hinting that the cgCaM gene is probably involved in shell formation of Pacific oyster, and the common ancestor of Gastropoda and Bivalvia may possess at least three CaM genes. We also found that the numbers of some EF hand family members in highly calcified species were higher than those in lowly calcified species and the numbers of these motifs in oyster genome were the highest among the mollusk species with whole genome sequence, further hinting the correlation between CaM and biomineralization.

  7. Molecular Cloning and Characterization of Full-Length cDNA of Calmodulin Gene from Pacific Oyster Crassostrea gigas

    Science.gov (United States)

    Li, Xing-Xia; Yu, Wen-Chao; Cai, Zhong-Qiang; He, Cheng; Wei, Na

    2016-01-01

    The shell of the pearl oyster (Pinctada fucata) mainly comprises aragonite whereas that of the Pacific oyster (Crassostrea gigas) is mainly calcite, thereby suggesting the different mechanisms of shell formation between above two mollusks. Calmodulin (CaM) is an important gene for regulating the uptake, transport, and secretion of calcium during the process of shell formation in pearl oyster. It is interesting to characterize the CaM in oysters, which could facilitate the understanding of the different shell formation mechanisms among mollusks. We cloned the full-length cDNA of Pacific oyster CaM (cgCaM) and found that the cgCaM ORF encoded a peptide of 113 amino acids containing three EF-hand calcium-binding domains, its expression level was highest in the mantle, hinting that the cgCaM gene is probably involved in shell formation of Pacific oyster, and the common ancestor of Gastropoda and Bivalvia may possess at least three CaM genes. We also found that the numbers of some EF hand family members in highly calcified species were higher than those in lowly calcified species and the numbers of these motifs in oyster genome were the highest among the mollusk species with whole genome sequence, further hinting the correlation between CaM and biomineralization. PMID:27703977

  8. Molecular cloning and expression of a cDNA encoding a hybrid histidine kinase receptor in tropical periwinkle Catharanthus roseus.

    Science.gov (United States)

    Papon, N; Bremer, J; Vansiri, A; Glévarec, G; Rideau, M; Creche, J

    2006-09-01

    Signalling pathways involving histidine kinase receptors (HKRs) are widely used by prokaryotes and fungi to regulate a large palette of biological processes. In plants, HKRs are known to be implicated in cytokinin, ethylene, and osmosensing transduction pathways. In this work, a full length cDNA named CRCIK was isolated from the tropical species CATHARANTHUS ROSEUS (L.) G. Don. It encodes a 1205 amino acid protein that belongs to the hybrid HKR family. The deduced amino acid sequence shows the highest homology with AtHK1, an osmosensing HKR in ARABIDOPSIS THALIANA. In return, CrCIK protein shares very low identity with the other 10 ARABIDOPSIS HKRs. Southern blot analysis indicates that the CRCIK corresponding gene is either present in multiple copies or has very close homologues in the genome of the tropical periwinkle. The gene is widely expressed in the plant. In C. ROSEUS C20D cell suspension, it is slightly induced after exposure to low temperature, pointing to a putative role in cold-shock signal transduction.

  9. Cloning and Characterization of Full Length cDNA of a CC-NBS-LRR Resistance Gene in Sweetpotato

    Institute of Scientific and Technical Information of China (English)

    CHEN Guan-shui; ZHOU Yi-fei; HOU Li-li; PAN Da-ren

    2009-01-01

    Conserved domain such as nucleotide binding site (NBS) was found in several cloned plant disease resistance genes.Based on the NBS domain,resistance gene analogues (RGAs) have been isolated.A full-length cDNA,SPRI was obtained by rapid amplification of cDNA ends (RACE) method.Sequence analysis indicated that the length of SPR1 was 3 066 bp,including a complete open reading frame of 2 667 bp encoding SPRI protein of 888 amino acids.Compared with known NBS-LRR genes,it presented relatively high amino acid sequence identity.The polypeptide has a typical structure of non TIR-NBS-LRR genes,with NB-ARC,CC,and LRR domains.The SPR1-related sequences belonged to multicopy gene family in sweetpotato genome according to the result of Southern blotting.Semi-quantitative RT-PCR analysis showed SPR1 expressed in all tested tissues.The cloning of putative resistance gene from sweetpotato provides a basis for studying the structure and function of sweetpotato disease-resistance relating genes and disease resistant genetic breeding in sweetpotato.The gene has been submitted to the GenBank database,and the accession number is EF428453.

  10. Construction of a cDNA library for sea cucumber Acaudina leucoprocta and differential expression of ferritin peptide

    Science.gov (United States)

    Zhou, Jun; Hou, Fujing; Li, Ye; Su, Xiurong; Li, Taiwu; Jin, Chunhua

    2016-07-01

    Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would contribute to a better understanding of the optimal conditions for its aquaculture and its mechanisms of defense against disease. Therefore, we constructed a cDNA library and, based on bioinformatics analysis of the sequences, the functions of 75% of the cDNAs were identified, including those involved in cell structure, energy metabolism, mitochondrial function, and signal transduction pathways. Approximately 25% of genes in the library were unmatched. The gene for A. leucoprocta ferritin was also cloned. The predicted amino-acid sequence of ferritin displayed significant homology with other sea-cucumber counterparts but indicated that it was a new member of the ferritin family. Semiquantitative real-time RT-PCR indicated the highest levels of ferritin mRNA expression in the intestine. A polyclonal antibody of ferritin was also produced. These data provide a set of molecular tools essential for further studies of the functions of ferritin protein in A. leucoprocta.

  11. Analysis of immune system gene expression in small rheumatoid arthritis biopsies using a combination of subtractive hybridization and high-density cDNA arrays.

    Science.gov (United States)

    Zanders, E D; Goulden, M G; Kennedy, T C; Kempsell, K E

    2000-01-13

    Subtractive hybridization of cDNAs generated from synovial RNA which had been isolated from patients with rheumatoid arthritis (RA) or normal controls was used in conjunction with high-density array hybridization to identify genes of immunological interest. The method was designed to detect gene expression in small needle biopsy specimens by means of a prior amplification of nanogram amounts of total RNA to full-length cDNA using PCR. The latter was cut with Rsa I, ligated with adapters, hybridized with unmodified driver cDNA, and subjected to suppression subtraction PCR. Differentially expressed products were cloned into E. coli and picked into 384 well plates. Inserts were obtained by PCR across the multiple cloning site, and the products arrayed at high density on nylon filters. The subtracted cDNAs were also labelled by random priming for use as probes for library screening. The libraries chosen were the subtracted one described above and a set of 45,000 ESTs from the I.M. A.G.E consortium. Clones showing positive hybridization were identified by sequence analysis and homology searching. The results showed that the subtracted hybridization approach could identify many gene fragments expressed at different levels, the most abundant being immunoglobulins and HLA-DR. The expression profile was characteristic of macrophage, B cell and plasma cell infiltration with evidence of interferon induction. In addition, a significant number of sequences without matches in the nucleotide databases were obtained, this demonstrates the utility of the method in finding novel gene fragments for further characterisation as potential members of the immune system. Although RA was studied here, the technology is applicable to any disease process even in cases where amounts of tissue may be limited.

  12. Familial dermographism.

    Science.gov (United States)

    Jedele, K B; Michels, V V

    1991-05-01

    Urticaria in response to various physical stimuli has been reported in sporadic and familial patterns. The most common of these physical urticarias, dermographism, is a localized urticarial response to stroking or scratching of the skin and has not been reported previously to be familial. A four-generation family with dermographism, probably inherited as an autosomal dominant trait, is presented along with a discussion of sporadic dermographism and other types of familial physical urticarias.

  13. Molecular cloning and characterization of a tomato cDNA encoding a systemically wound-inducible bZIP DNA-binding protein

    Science.gov (United States)

    Stankovic, B.; Vian, A.; Henry-Vian, C.; Davies, E.

    2000-01-01

    Localized wounding of one leaf in intact tomato (Lycopersicon esculentum Mill.) plants triggers rapid systemic transcriptional responses that might be involved in defense. To better understand the mechanism(s) of intercellular signal transmission in wounded tomatoes, and to identify the array of genes systemically up-regulated by wounding, a subtractive cDNA library for wounded tomato leaves was constructed. A novel cDNA clone (designated LebZIP1) encoding a DNA-binding protein was isolated and identified. This clone appears to be encoded by a single gene, and belongs to the family of basic leucine zipper domain (bZIP) transcription factors shown to be up-regulated by cold and dark treatments. Analysis of the mRNA levels suggests that the transcript for LebZIP1 is both organ-specific and up-regulated by wounding. In wounded wild-type tomatoes, the LebZIP1 mRNA levels in distant tissue were maximally up-regulated within only 5 min following localized wounding. Exogenous abscisic acid (ABA) prevented the rapid wound-induced increase in LebZIP1 mRNA levels, while the basal levels of LebZIP1 transcripts were higher in the ABA mutants notabilis (not), sitiens (sit), and flacca (flc), and wound-induced increases were greater in the ABA-deficient mutants. Together, these results suggest that ABA acts to curtail the wound-induced synthesis of LebZIP1 mRNA.

  14. Cloning of the cDNA for a human homologue of the Drosophila white gene and mapping to chromosome 21q22.3

    Energy Technology Data Exchange (ETDEWEB)

    Haiming Chen; Lalioti, M.D.; Perrin, G.; Antonarakis, S.E. [Univ. of Geneva Medical School (Switzerland)] [and others

    1996-07-01

    In an effort to contribute to the transcript map of human chromosome 21 and the understanding of the pathophysiology of trisomy 21, we have used exon trapping to identify fragments of chromosome 21 genes. Two trapped exons, from pools of chromosome 21-specific cosmids, showed homology to the Drosophila white (w) gene. We subsequently cloned the corresponding cDNA for a human homologue of the Drosophila w gene (hW) from human retina and fetal brain cDNA libraries. The gene belongs to the ATP-binding cassette transporter gene family and is homologous to Drosophila w (and to 2 genes from other species) and to a lesser extent to Drosophila brown (bw) and scarlet (st) genes that are all involved in the transport of eye pigment precursor molecules. A DNA polymorphism with 62% heterozygosity due to variation of a poly (T) region in the 3{prime} UTR of the hW has been identified and used for the incorporation of this gene to the genetic map of chromosome 21. The hW is located at 21q22.3 between DNA markers D21S212 and D21S49 in a P1 clone that also contains marker BCEI. The gene is expressed at various levels in many human tissues. The contributions of this gene to the Down syndrome phenotypes, to human eye color, and to the resulting phenotypes of null or missense mutations are presently unknown. 56 refs., 8 figs., 1 tab.

  15. cDNA sequence, gene structure, and cholinesterase-like domains of an esterase from Caenorhabditis elegans mapped to chromosome V.

    Science.gov (United States)

    Fedon, Y; Cousin, X; Toutant, J P; Thierry-Mieg, D; Arpagaus, M

    1993-01-01

    The structure of an esterase gene from Caenorhabditis elegans has been determined by comparison of the sequences in genomic and cDNA clones. The gene was mapped close to the center of chromosome V (1.7 centimorgans to the left of dpy-11) and is therefore distinct from the gut esterase gene ges-1. It possessed 7 short introns. The 5' splice site of intron 3 presented the sequence GC instead of the usual GT that was found in the other six introns. The cDNA was trans-spliced with the short leader SL1. The open reading frame indicated that a protein of 557 aminoacids was encoded. The deduced aminoacid sequence did not present a signal peptide at the N-terminal but a potential N-myristoylation site (GXXXS) provided that the initiator methionine was removed. This protein should therefore remain intracellular. Comparison of this C. elegans sequence to other protein sequences in databases, as well as the analysis of the secondary structure in the protein showed that it belongs to the subgroup of esterases in the alpha/beta hydrolase fold family.

  16. cDNA sequence, gene structure, and in vitro expression of ace-1, the gene encoding acetylcholinesterase of class A in the nematode Caenorhabditis elegans.

    Science.gov (United States)

    Arpagaus, M; Fedon, Y; Cousin, X; Chatonnet, A; Bergé, J B; Fournier, D; Toutant, J P

    1994-04-01

    Three genes, ace-1, ace-2, and ace-3, encode three acetylcholinesterase classes (A, B, and C) in the nematode Caenorhabditis elegans. A fragment of genomic DNA was amplified by a polymerase chain reaction (PCR) using degenerate oligonucleotides based on sequences conserved in the cholinesterase family. This fragment mapped to chromosome X at a position that perfectly matched the location of ace-1 previously determined by genetic methods. Comparison of genomic and cDNA sequences showed that the open reading frame was interrupted by eight introns. The product of ace-1 (ACE-1, 620 amino acids) presented 42% identity with Torpedo and human acetylcholinesterases, 41% with human butyrylcholinesterase, and 35% with Drosophila acetylcholinesterase. The overall structure of cholinesterases was conserved in ACE-1 as indicated by the conserved sequence positions of Ser-216, His-468, and Glu-346 (S200, H440, E327 in Torpedo (AChE) as components of the catalytic triad, of the six cysteines which form three intrachain disulfide bonds, and of Trp-99(84), a critical side chain in the choline binding site. Spodoptera Sf9 cells were infected by a recombinant baculovirus containing ace-1 cDNA. The secreted enzyme was active and existed as hydrophilic 5 and 11.5 S molecular forms. It hydrolyzed both acetylthiocholine and butyrylthiocholine and was inhibited by acetylthiocholine above 10 mM.

  17. Cloning of the 5' mRNA for the 230-kD bullous pemphigoid antigen by rapid amplification of cDNA ends.

    Science.gov (United States)

    Elgart, G W; Stanley, J R

    1993-08-01

    The 230-kD bullous pemphigoid antigen (BPAG1), defined by autoantibodies in patient sera, is a hemidesmosomal plaque protein in the same gene family as the intracellular proteins desmoplakin I/II and plectin. We had previously isolated, from a lambda gt11 library, overlapping cDNA clones with 6921 bp of mRNA sequence for BPAG1. The coding sequence encoded by these clones included the 3' stop codon but not the 5' coding and non-coding region of the mRNA. To obtain these sequences we used the polymerase chain reaction (PCR) method called rapid amplification of cDNA ends (RACE). The PCR products were cloned into plasmids and sequenced. With five PCR primers we were able to obtain overlapping clones containing the 5' region of the mRNA. An upstream stop codon in frame with the rest of the coding sequence demonstrates that the full 5' coding sequence is obtained. Four different PCR products from two separate reactions had the same 5' end, suggesting that this 5' end is near, or at, the transcription start site. No alternatively spliced clones were found and no transmembrane site was predicted, confirming that BPAG1 is an intracellular hemidesmosomal plaque protein.

  18. Molecular cloning and characterization of a tomato cDNA encoding a systemically wound-inducible bZIP DNA-binding protein

    Science.gov (United States)

    Stankovic, B.; Vian, A.; Henry-Vian, C.; Davies, E.

    2000-01-01

    Localized wounding of one leaf in intact tomato (Lycopersicon esculentum Mill.) plants triggers rapid systemic transcriptional responses that might be involved in defense. To better understand the mechanism(s) of intercellular signal transmission in wounded tomatoes, and to identify the array of genes systemically up-regulated by wounding, a subtractive cDNA library for wounded tomato leaves was constructed. A novel cDNA clone (designated LebZIP1) encoding a DNA-binding protein was isolated and identified. This clone appears to be encoded by a single gene, and belongs to the family of basic leucine zipper domain (bZIP) transcription factors shown to be up-regulated by cold and dark treatments. Analysis of the mRNA levels suggests that the transcript for LebZIP1 is both organ-specific and up-regulated by wounding. In wounded wild-type tomatoes, the LebZIP1 mRNA levels in distant tissue were maximally up-regulated within only 5 min following localized wounding. Exogenous abscisic acid (ABA) prevented the rapid wound-induced increase in LebZIP1 mRNA levels, while the basal levels of LebZIP1 transcripts were higher in the ABA mutants notabilis (not), sitiens (sit), and flacca (flc), and wound-induced increases were greater in the ABA-deficient mutants. Together, these results suggest that ABA acts to curtail the wound-induced synthesis of LebZIP1 mRNA.

  19. A Dictatorship Theorem for Cake Cutting

    DEFF Research Database (Denmark)

    Branzei, Simina; Miltersen, Peter Bro

    2015-01-01

    We consider discrete protocols for the classical Steinhaus cake cutting problem. Under mild technical conditions, we show that any deterministic strategy-proof protocol in the standard Robertson-Webb query model is dictatorial, that is, there is a fixed agent to which the protocol allocates...

  20. Indirect Revelation Mechanisms for Cake Cutting

    DEFF Research Database (Denmark)

    Branzei, Simina; Miltersen, Peter Bro

    We consider discrete protocols for the classical Steinhaus cake cutting problem. Under mild technical conditions, we show that any deterministic strategy-proof protocol in the standard Robertson-Webb query model is dictatorial, that is, there is a fixed agent to which the protocol allocates...