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Sample records for dos clones gt

  1. [Cloning and functional analysis of the cotton Trihelix transcription factor GhGT29].

    PubMed

    Yue, Li; Xiaodong, Liu; Yongmei, Dong; Zongming, Xie; Shouyi, Chen

    2015-12-01

    Trihelix transcription factors are important proteins involved in response to abiotic stresses in plants. Understanding the molecular mechanisms of Trihelix in cottons will lay the foundation to improve stress tolerance by gene engineering. In this study, a gene encoding Trihelix transcription factor was isolated in upland cottons using reverse transcription PCR according to bioinformatic analysis. The gene was named as GhGT29 (GenBank accession No. JQ013097), which was 1 092 bp, contained a 1 089 bp open reading frame and encoded a protein of 363 amino acids with a predicted molecular weight of 40.9 kDa and a isoelectric point of 5.45. SMART analysis showed GhGT29 contained one typical SANT motif. Phylogenetic analysis showed that GhGT29 belonged to the SH4 subfamily of the Trihelix family and was most closely related to AtSH4-like1 and AtSH4-like2. Quantitative real-time PCR (qRT-PCR) analysis revealed that GhGT29 was induced by high salt, drought, cold and abscisic acid. The expression profile also revealed that GhGT29 was constitutively expressed in all tested tissues, such as roots, stems, leaves, flowers, ovules (0 DPA) and fibers (12 DPA). The expression level of GhGT29 was the highest in flowers and the lowest in stems. Using the Arabidopsis protoplasts assay system, we found that the GhGT29 protein was located in cell nuclei and had trans-activation activity. These results revealed that GhGT29 might be involved in the regulation of stress resistance-related genes in stress signaling pathways in upland cottons.

  2. Cloning

    MedlinePlus

    Cloning describes the processes used to create an exact genetic replica of another cell, tissue or organism. ... named Dolly. There are three different types of cloning: Gene cloning, which creates copies of genes or ...

  3. Cloning

    MedlinePlus

    ... 2001, researchers produced the first clone of an endangered species: a type of Asian ox known as a ... few days after its birth. In 2003, another endangered type of ox, called the ... many species that would otherwise disappear, others argue that cloning ...

  4. Rice Glycosyltransferase (GT) Phylogenomic Database

    DOE Data Explorer

    Ronald, Pamela

    The Ronald Laboratory staff at the University of California-Davis has a primary research focus on the genes of the rice plant. They study the role that genetics plays in the way rice plants respond to their environment. They created the Rice GT Database in order to integrate functional genomic information for putative rice Glycosyltransferases (GTs). This database contains information on nearly 800 putative rice GTs (gene models) identified by sequence similarity searches based on the Carbohydrate Active enZymes (CAZy) database. The Rice GT Database provides a platform to display user-selected functional genomic data on a phylogenetic tree. This includes sequence information, mutant line information, expression data, etc. An interactive chromosomal map shows the position of all rice GTs, and links to rice annotation databases are included. The format is intended to "facilitate the comparison of closely related GTs within different families, as well as perform global comparisons between sets of related families." [From http://ricephylogenomics.ucdavis.edu/cellwalls/gt/genInfo.shtml] See also the primary paper discussing this work: Peijian Cao, Laura E. Bartley, Ki-Hong Jung and Pamela C. Ronalda. Construction of a Rice Glycosyltransferase Phylogenomic Database and Identification of Rice-Diverged Glycosyltransferases. Molecular Plant, 2008, 1(5): 858-877.

  5. Family 34 glycosyltransferase (GT34) genes and proteins in Pinus radiata (radiata pine) and Pinus taeda (loblolly pine).

    PubMed

    Ade, Carsten P; Bemm, Felix; Dickson, James M J; Walter, Christian; Harris, Philip J

    2014-04-01

    Using a functional genomics approach, four candidate genes (PtGT34A, PtGT34B, PtGT34C and PtGT34D) were identified in Pinus taeda. These genes encode CAZy family GT34 glycosyltransferases that are involved in the synthesis of cell-wall xyloglucans and heteromannans. The full-length coding sequences of three orthologs (PrGT34A, B and C) were isolated from a xylem-specific cDNA library from the closely related Pinus radiata. PrGT34B is the ortholog of XXT1 and XXT2, the two main xyloglucan (1→6)-α-xylosyltransferases in Arabidopsis thaliana. PrGT34C is the ortholog of XXT5 in A. thaliana, which is also involved in the xylosylation of xyloglucans. PrGT34A is an ortholog of a galactosyltransferase from fenugreek (Trigonella foenum-graecum) that is involved in galactomannan synthesis. Truncated coding sequences of the genes were cloned into plasmid vectors and expressed in a Sf9 insect cell-culture system. The heterologous proteins were purified, and in vitro assays showed that, when incubated with UDP-xylose and cellotetraose, cellopentaose or cellohexaose, PrGT34B showed xylosyltransferase activity, and, when incubated with UDP-galactose and the same cello-oligosaccharides, PrGT34B showed some galactosyltransferase activity. The ratio of xylosyltransferase to galactosyltransferase activity was 434:1. Hydrolysis of the galactosyltransferase reaction products using galactosidases showed the linkages formed were α-linkages. Analysis of the products of PrGT34B by MALDI-TOF MS showed that up to three xylosyl residues were transferred from UDP-xylose to cellohexaose. The heterologous proteins PrGT34A and PrGT34C showed no detectable enzymatic activity. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  6. Asymmetric GT of social networks

    NASA Astrophysics Data System (ADS)

    Szu, Harold

    2010-04-01

    Web citation indexes are computed according to a data vector X collected from the frequency of user accesses, citations weighted by other sites' popularities, and modified by the financial sponsorship in a proprietary manner. The indexing determining the information to be retrieved by the public should be made responsible transparently in at least two ways. One shall balance the inbound linkages pointed at the specific i-th site called the popularity (see paper for equation) with the outbound linkages (see paper for equation) called the risk factor before the release of new information as environmental impact analysis. The relationship between these two factors cannot be assumed equivalent (undirected) as in the case of many mainstream Graph Theory (GT) models.

  7. Registration of maize inbred line 'GT888'

    USDA-ARS?s Scientific Manuscript database

    Maize (Zea mays L.) inbred line GT888 (PI 670116) was developed and released by the USDA-ARS in cooperation with the University of Georgia, and in participation with the USDA Germplasm Enhancement of Maize (GEM) project. GT888 was derived from GEM population DK888:N11 (GEMN-0177), which has 50% tro...

  8. Registration of maize inbred line GT603

    USDA-ARS?s Scientific Manuscript database

    GT603 (Reg. No. xxxx, PI xxxxxx) is a yellow dent maize (Zea mays L.) inbred line developed and released by the USDA-ARS Crop Protection and Management Research Unit in cooperation with the University of Georgia Coastal Plain Experiment Station in 2010. GT603 was developed through seven generations ...

  9. GT Strength in Odd-A Nuclei^*

    NASA Astrophysics Data System (ADS)

    Watson, J. W.; Du, Q. Q.

    1998-04-01

    We measured the complete set of polarization-transfer observables (D_ij) for the ^13C(p,n)^13N and ^15N(p,n)^15O reactions at 135 MeV. From the D_ijs we constructed the spin-independent, spin-longitudinal, and spin-transverse responses [1] D_0, D_q, Dn and D_p. From these responses we extracted, in a model-independent way, the Δ J=0 and Δ J=1 (``F'' and ``GT'') fractions of the J^π=1/2^-arrow1/2^- g.s. to g.s. transitions for these reactions. The ``F'' fraction, f_F=D_0(0^circ); the ``GT'' fraction, f_GT=D_q(0^circ)+D_n(0^circ)+D_p(0^circ)= 1- D_0(0^circ). The extracted GT fractions, f_GT, are substantially larger than one would predict from β-decay matrix elements and the systematics of the (p,n) reaction on even-A nuclei. These results confirm earlier, model-dependent determinations of f_GT obtained from the (p,n) reaction on ^13C, ^15N and ^39K at other energies [2], [3], [4], indicating that considerable caution must be used when extracting B(GT) matrix elements from odd-A (p,n) data. * Research supported in part by the U.S. NSF. [1] M. Ichimura, K. Kawahigashi, Phys. Rev. C 45 1822 (1992). [2] T. N. Taddeucci, C. A. Goulding, T. A. Carey, R. C. Byrd, C. D. Goodman, C. Gaarde, J. Larsen, D. Horen, J. Rapaport, and E. Sugarbaker, Nucl. Phys. A469 125 (1987). [3] H. Sakai, H. Okamura, N. Matsuoka, A. Shimizu, T. Suda, M. Ieiri and H. M. Shimizu, Nuclear Physics A579 45-61 (1994). [4] W. Huang, Ph.D. dissertation, Indiana U., 1991, (unpublished).

  10. GT Merge Process: Version 2.0

    SciTech Connect

    Flanagan, M P; Dodge, D; Myers, S C

    2008-06-10

    This document summarizes the process used to merge GT25 and better data between LANL and LLNL. The merge also includes OUO arrivals provided by AFTAC for events in the merge. The merge process is mostly automated and includes extensive quality control operations at each step. Events in common between the labs are identified and resolved using GT level criteria. Arrivals in common between the labs are also resolved through the use of agreed upon arrival author rankings. Finally, baselined origin times are computed for all crustal events using either teleseismic P-arrivals and the iasp91 model or, in certain regions, regional P-arrivals and regional velocity models that are known to be consistent with teleseismic iasp91 P-wave predictions. We combine the core tables from each contributor and resolve unique and common GT events between contributors. Next, we merge at the pick level so that each distinct EVENT-STATION-PHASE tuple has a unique arrival. All BMEB (Bondar-Myers-Engdahl-Bergman) GT are recalculated and evaluated for adherence to their criteria. Finally, new origin times are computed (baselining) for the merged GT events. In addition to the reconciliation of events and picks between contributors, the merge process involves several quality control steps that are intended to remove outlier and irrelevant data from the final results. The process is described in the section entitled 'Merge Steps'.

  11. GT Merge Process: Version 1.0

    SciTech Connect

    Flanagan, M P; Dodge, D; Myers, S C

    2008-06-10

    This document summarizes the process used to merge GT25 and better data between LANL and LLNL for use in a tomographic inversion for Pn velocity of Eurasia. The merge process is automated and includes extensive quality control operations at each step. Events in common between the labs are identified and resolved using GT level criteria. Arrivals in common between the labs are also resolved through the use of agreed upon arrival author rankings. Finally, baselined origin times are computed for all crustal events using either teleseismic P-arrivals and the iasp91 model or, in certain regions, regional P-arrivals and regional velocity models that are known to be consistent with teleseismic iasp91 P-wave predictions. We combine the core tables from each lab and first resolve unique and common GT events between LANL and LLNL. Phase names are then checked and possibly adjusted for consistency. Next, we merge at the pick level so that each distinct EVENT-STATION-PHASE tuple has a unique arrival. All BMEB (Bondar-Myers-Engdahl-Bergman) GT are evaluated for adherence to their criteria, and possibly re-calculated. Finally, new origin times are computed (baselining) for the merged GT events. In addition to the reconciliation of events and picks between LANL and LLNL, the merge process involves several quality control steps that are intended to remove outlier and irrelevant data from the final results.

  12. Comparative analysis of GT14/GT14-like family genes in Arabidopsis, Oryza, Populus, Sorghum and Vitis

    SciTech Connect

    Ye, Chuyu; Li, Ting; Tuskan, Gerald A; Tschaplinski, Timothy J; Yang, Xiaohan

    2011-01-01

    Glycosyltransferase family14 (GT14) belongs to the glycosyltransferase (GT) superfamily that plays important roles in the biosynthesis of cell walls, the most abundant source of cellulosic biomass for bioethanol production. It has been hypothesized that DUF266 proteins are a new class of GTs related to GT14. In this study, we identified 62 GT14 and 106 DUF266 genes (named GT14-like herein) in Arabidopsis, Oryza, Populus, Sorghum and Vitis. Our phylogenetic analysis separated GT14 and GT14-like genes into two distinct clades, which were further divided into eight and five groups, respectively. Similarities in protein domain, 3D structure and gene expression were uncovered between the two phylogenetic clades, supporting the hypothesis that GT14 and GT14-like genes belong to one family. Therefore, we proposed a new family name, GT14/GT14-like family that combines both subfamilies. Variation in gene expression and protein subcellular localization within the GT14-like subfamily were greater than those within the GT14 subfamily. One-half of the Arabidopsis and Populus GT14/GT14-like genes were found to be preferentially expressed in stem/xylem, indicating that they are likely involved in cell wall biosynthesis. This study provided new insights into the evolution and functional diversification of the GT14/GT14-like family genes.

  13. The Poplar GT8E and GT8F Glycosyltransferases are Functional Orthologs of Arabidopsis PARVUS Involved in Gulcuronoxylan Biosynthesis

    EPA Science Inventory

    The poplar GT8E and GT8F glycosyltransferases have previously been shown to be associated with wood formation, but their roles in the biosynthesis of wood components are not known. Here, we show that PoGT8E and PoGT8F are expressed in vessels and fibers during wood formation and ...

  14. The Poplar GT8E and GT8F Glycosyltransferases are Functional Orthologs of Arabidopsis PARVUS Involved in Gulcuronoxylan Biosynthesis

    EPA Science Inventory

    The poplar GT8E and GT8F glycosyltransferases have previously been shown to be associated with wood formation, but their roles in the biosynthesis of wood components are not known. Here, we show that PoGT8E and PoGT8F are expressed in vessels and fibers during wood formation and ...

  15. RECOVERY - GEMINI-TITAN (GT)-4 - ATLANTIC

    NASA Image and Video Library

    1965-06-07

    S65-33490 (7 June 1965) --- A United States Navy frogman team participates in the recovery of the Gemini-Titan 4 (GT-4) spacecraft. The USS Wasp was the prime recovery ship for the Gemini-4 mission. The crew of the Gemini-4 spaceflight was astronauts James A. McDivitt, command pilot, and Edward H. White II, pilot.

  16. Cloning and characterization of the UDP-glucose:anthocyanin 5-O-glucosyltransferase gene from blue-flowered gentian.

    PubMed

    Nakatsuka, Takashi; Sato, Kei; Takahashi, Hideyuki; Yamamura, Saburo; Nishihara, Masahiro

    2008-01-01

    Blue-flowered gentian (Gentiana triflora) is known to accumulate gentiodelphin, a unique polyacylated delphinidin-type anthocyanin, in the petals. Almost all of the structural genes involved in gentiodelphin biosynthesis have been isolated, but an important gene encoding UDP-glucose:anthocyanin 5-O-glucosyltransferase (5GT) remained to be identified. In this study, an attempt was made to isolate and characterize gentian 5GT, which is responsible for glucosylation of anthocyanidin 3-glucoside. A PCR-based cloning strategy identified seven 5GT candidates from gentian flowers. Among them, the deduced amino acid sequence of the 5GT gene from gentian petal cDNA, designated Gt5GT7, exhibited 36.0-41.7% identities with those of 5GTs from other plant species, and phylogenic analysis also suggested that Gt5GT7 belongs to the 5GT subfamily. The expression analysis showed that Gt5GT7 transcripts were detected predominantly in petals and weakly in filaments but not in leaves, stems, and other floral organs. In addition, increased levels of Gt5GT7 transcripts in petals coincided with flower development, a pattern identical to that of 5GT enzymatic activity as determined by in vitro assay using petal crude proteins. The substrate specificity of Gt5GT7 was analysed in vitro using the recombinant enzyme produced by Escherichia coli. Gt5GT7 could transfer a glucosyl moiety to anthocyanidin 3-glycosides but not to other flavonoid compounds. Delphinidin 3-glucoside, the precursor of gentiodelphin, was the best substrate among several anthocyanidin 3-glycosides tested. Heterologous expression of Gt5GT7 in tobacco plants led to additional accumulation of cyanidin 3-rutinoside-5-glucoside, confirming that Gt5GT7 has a valid enzymatic activity in planta.

  17. The Wheat GT Factor TaGT2L1D Negatively Regulates Drought Tolerance and Plant Development

    PubMed Central

    Zheng, Xin; Liu, Haipei; Ji, Hongtao; Wang, Youning; Dong, Baodi; Qiao, Yunzhou; Liu, Mengyu; Li, Xia

    2016-01-01

    GT factors are trihelix transcription factors that specifically regulate plant development and stress responses. Recently, several GT factors have been characterized in different plant species; however, little is known about the role of GT factors in wheat. Here, we show that TaGT2L1A, TaGT2L1B, and TaGT2L1D are highly homologous in hexaploid wheat, and are localized to wheat chromosomes 2A, 2B, and 2D, respectively. These TaGT2L1 genes encode proteins containing two SANT domains and one central helix. All three homologs were ubiquitously expressed during wheat development and were responsive to osmotic stress. Functional analyses demonstrated that TaGT2L1D acts as a transcriptional repressor; it was able to suppress the expression of AtSDD1 in Arabidopsis by binding directly to the GT3 box in its promoter that negatively regulates drought tolerance. TaGT2L1D overexpression markedly increased the number of stomata and reduced drought tolerance in gtl1-3 plants. Notably, ectopic expression of TaGT2L1D also affected floral organ development and overall plant growth. These results demonstrate that TaGT2L1 is an ortholog of AtGTL1, and that it plays an evolutionarily conserved role in drought resistance by fine tuning stomatal density in wheat. Our data also highlight the role of TaGT2L1 in plant growth and development. PMID:27245096

  18. High antagonist potency of GT-2227 and GT-2331, new histamine H3 receptor antagonists, in two functional models.

    PubMed

    Tedford, C E; Hoffmann, M; Seyedi, N; Maruyama, R; Levi, R; Yates, S L; Ali, S M; Phillips, J G

    1998-06-26

    GT-2227 (4-(6-cyclohexylhex-cis-3-enyl)imidazole) and GT-2331 ((1R,2R)-4-(2-(5,5-dimethylhex-1-ynyl)cyclopropyl)imidazole) were developed as new potent histamine H3 receptor antagonists. The functional activity of these ligands on the histamine H3 receptor-mediated inhibition of neurogenic contraction of the guinea-pig jejunum and histamine H3 receptor-mediated inhibition of norepinephrine release from guinea-pig heart synaptosomes were investigated. GT-2227 and GT-2331 both antagonized the inhibitory effects of (R)-alpha-methylhistamine on the contraction induced by electrical field stimulation in the guinea-pig jejunum with pA2 values of 7.9+/-0.1 and 8.5+/-0.03, respectively. In addition, GT-2227 and GT-2331 antagonized the inhibition of norepinephrine release in cardiac synaptosomes by GT-2203 ((1R,2R)-trans-2-(1H-imidazol-4-yl)cyclopropylamine), a histamine H3 receptor agonist. The current results demonstrate the antagonist activity for both GT-2227 and GT-2331 in two functional assays for histamine H3 receptors.

  19. Technical Reliability Assessment of the Actigraph GT1M Accelerometer

    ERIC Educational Resources Information Center

    Silva, Pedro; Mota, Jorge; Esliger, Dale; Welk, Gregory

    2010-01-01

    The purpose of this study was to determine the reliability of the Actigraph GT1M (Pensacola, FL, USA) accelerometer activity count and step functions. Fifty GT1M accelerometers were initialized to collect simultaneous acceleration counts and steps data using 15-sec epochs. All reliability testing was completed using a mechanical shaker plate to…

  20. Technical Reliability Assessment of the Actigraph GT1M Accelerometer

    ERIC Educational Resources Information Center

    Silva, Pedro; Mota, Jorge; Esliger, Dale; Welk, Gregory

    2010-01-01

    The purpose of this study was to determine the reliability of the Actigraph GT1M (Pensacola, FL, USA) accelerometer activity count and step functions. Fifty GT1M accelerometers were initialized to collect simultaneous acceleration counts and steps data using 15-sec epochs. All reliability testing was completed using a mechanical shaker plate to…

  1. Cloning cattle.

    PubMed

    Oback, B; Wells, D N

    2003-01-01

    Over the past six years, hundreds of apparently normal calves have been cloned worldwide from bovine somatic donor cells. However, these surviving animals represent less than 5% of all cloned embryos transferred into recipient cows. Most of the remaining 95% die at various stages of development from a predictable pattern of placental and fetal abnormalities, collectively referred to as the "cloning-syndrome." The low efficiency seriously limits commercial applicability and ethical acceptance of somatic cloning and enforces the development of improved cloning methods. In this paper, we describe our current standard operating procedure (SOP) for cattle cloning using zona-free nuclear transfer. Following this SOP, the output of viable and healthy calves at weaning is about 9% of embryos transferred. Better standardization of cloning protocols across and within research groups is needed to separate technical from biological factors underlying low cloning efficiency.

  2. 3. AIR TO GROUND RADAR TYPE GT2122 & GRRR 2324, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    3. AIR TO GROUND RADAR TYPE GT2122 & GRRR 2324, CIRCA 1978, INTERIOR OF BUILDING 408, LOOKING WEST. - Mill Valley Air Force Station, Operations Building & Annex, East Ridgecrest Boulevard, Mount Tamalpais, Mill Valley, Marin County, CA

  3. [Cloning - controversies].

    PubMed

    Twardowski, T; Michalska, A

    2001-01-01

    Cloning of the human being is not only highly controversial; in the opinion of the authors it is impossible - we are not able to reproduce human behaviour and character traits. Reproduction through cloning is limited to personal genome resources. The more important is protection of genomic characteristics as private property and taking advantage of cloning for production of the human organs directly or through xenotransplants. In this paper we present the legislation related to cloning in Poland, in the European Union and other countries. We also indicate who and why is interested in cloning.

  4. Two flavonoid glucosyltransferases from Petunia hybrida: molecular cloning, biochemical properties and developmentally regulated expression.

    PubMed

    Yamazaki, Mami; Yamagishi, Emiko; Gong, Zhizhong; Fukuchi-Mizutani, Masako; Fukui, Yuko; Tanaka, Yoshikazu; Kusumi, Takaaki; Yamaguchi, Masaatsu; Saito, Kazuki

    2002-03-01

    Two flavonoid glucosyltransferases, UDP-glucose:flavonoid 3-0-glucosyltransferase (3-GT) and UDP-glucose: anthocyanin 5-O-glucosyltransferase (5-GT), are responsible for the glucosylation of anthocyani(di)ns to produce stable molecules in the anthocyanin biosynthetic pathway. The cDNAs encoding 3-GT and 5-GT were isolated from Petunia hybrida by hybridization screening with heterologous probes. The cDNA clones of 3-GT, PGT8, and 5-GT, PH1, encode putative polypeptides of 448 and 468 amino acids, respectively. A phylogenetic tree based on amino acid sequences of the family of glycosyltransferases from various plants shows that PGT8 belongs to the 3-GT subfamily and PH1 belongs to the 5-GT subfamily. The function of isolated cDNAs was identified by the catalytic activities for 3-GT and 5-GT exhibited by the recombinant proteins produced in yeast. The recombinant PGT8 protein could convert not only anthocyanidins but also flavonols into the corresponding 3-O-glucosides. In contrast, the recombinant PH1 protein exhibited a strict substrate specificity towards anthocyanidin 3-acylrutinoside, comparing with other 5-GTs from Perilla frutescens and Verbena hybrida, which showed broad substrate specificities towards several anthocyanidin 3-glucosides. The mRNA expression of both 3-GT and 5-GT increased in the early developmental stages of P. hybrida flower, reaching the maximum at the stage before flower opening. Southern blotting analysis of genomic DNA indicates that both 3-GT and 5-GT genes exist in two copies in P. hybrida, respectively. The results are discussed in relation to the molecular evolution of flavonoid glycosyltransferases.

  5. Characterisation of Muta™Mouse λgt10-lacZ transgene: evidence for in vivo rearrangements

    PubMed Central

    Shwed, Philip S.; Crosthwait, Jennifer; Douglas, George R.; Seligy, Vern L.

    2010-01-01

    The multicopy λgt10-lacZ transgene shuttle vector of Muta™Mouse serves as an important tool for genotoxicity studies. Here, we describe a model for λgt10-lacZ transgene molecular structure, based on characterisation of transgenes recovered from animals of our intramural breeding colony. Unique nucleotide sequences of the 47 513 bp monomer are reported with GenBank® assigned accession numbers. Besides defining ancestral mutations of the λgt10 used to construct the transgene and the Muta™Mouse precursor (strain 40.6), we validated the sequence integrity of key λ genes needed for the Escherichia coli host-based mutation reporting assay. Using three polymerase chain reaction (PCR)-based chromosome scanning and cloning strategies, we found five distinct in vivo transgene rearrangements, which were common to both sexes, and involved copy fusions generating ∼10 defective copies per haplotype. The transgene haplotype was estimated by Southern hybridisation and real-time–polymerase chain reaction, which yielded 29.0 ± 4.0 copies based on spleen DNA of Muta™Mouse, and a reconstructed CD2F1 genome with variable λgt10-lacZ copies. Similar analysis of commercially prepared spleen DNA from Big Blue® mouse yielded a haplotype of 23.5 ± 3.1 copies. The latter DNA is used in calibrating a commercial in vitro packaging kit for E.coli host-based mutation assays of both transgenic systems. The model for λgt10-lacZ transgene organisation, and the PCR-based methods for assessing copy number, integrity and rearrangements, potentially extends the use of Muta™Mouse construct for direct, genomic-type assays that detect the effects of clastogens and aneugens, without depending on an E.coli host, for reporting effects. PMID:20724577

  6. Human Cloning

    DTIC Science & Technology

    2006-07-20

    not believe that noncoital, asexual reproduction , such as cloning, would be considered a fundamental right by the Supreme Court. A ban on human...society by “crossing the boundary from sexual to asexual reproduction , thus approving in principle the genetic manipulation and control of nascent human... reproductive cloning and, by a vote of 10 to 7, a four-year moratorium on cloning for medical research purposes. The ethical issues surrounding reproductive

  7. HTGR-GT and electrical load integrated control

    SciTech Connect

    Chan, T.; Openshaw, F.; Pfremmer, D.

    1980-05-01

    A discussion of the control and operation of the HTGR-GT power plant is presented in terms of its closely coupled electrical load and core cooling functions. The system and its controls are briefly described and comparisons are made with more conventional plants. The results of analyses of selected transients are presented to illustrate the operation and control of the HTGR-GT. The events presented were specifically chosen to show the controllability of the plant and to highlight some of the unique characteristics inherent in this multiloop closed-cycle plant.

  8. PRESS CONFERENCE - GEMINI-TITAN (GT)-3 - FL

    NASA Image and Video Library

    1965-03-25

    S65-20864 (25 March 1965) --- News conference held at the Carriage House press site the day after the successful Gemini-Titan 3 three-orbit mission. Being interviewed at the press table by news media are (left to right) Dr. Kurt H. Debus, director of Kennedy Space Center; Christopher C. Kraft Jr., MSC assistant director for Flight Operations; astronaut John W. Young, pilot of the GT-3 flight; astronaut Virgil I. Grissom, command pilot of the GT-3 mission; Dr. Robert R. Gilruth, MSC director; Dr. Robert C. Seamans, NASA associate administrator; and Julian Scheer, assistant administrator, Office Of Public Affairs, NASA.

  9. Academic Cloning.

    ERIC Educational Resources Information Center

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

    The authors define "cloning" as an integral feature of all educational systems, citing teaching practices which reward students for closely reproducing the teacher's thoughts and/or behaviors and administrative systems which tend to promote like-minded subordinates. They insist, however, that "academic cloning" is not a totally…

  10. Academic Cloning.

    ERIC Educational Resources Information Center

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

    The authors define "cloning" as an integral feature of all educational systems, citing teaching practices which reward students for closely reproducing the teacher's thoughts and/or behaviors and administrative systems which tend to promote like-minded subordinates. They insist, however, that "academic cloning" is not a totally…

  11. Over-expression of a putative poplar glycosyltransferase gene, PtGT1, in tobacco increases lignin content and causes early flowering

    PubMed Central

    Wang, Yan-Wen; Wang, Wen-Chao; Jin, Shang-Hui; Wang, Jun; Wang, Bo; Hou, Bing-Kai

    2012-01-01

    Family 1 glycosyltransferases catalyse the glycosylation of small molecules and play an important role in maintaining cell homeostasis and regulating plant growth and development. In this study, a putative glycosyltransferase gene of family 1, PtGT1, was cloned from poplar (Populus tomentosa Carr.). Sequence analysis showed that this gene encodes a protein of 481 amino acid residues with a conserved PSPG box at its C-terminal, suggesting that it is active in the glycosylation of plant secondary products. The PtGT1 gene was expressed in poplar stems and leaves, with a particularly high expression level in elongating stems. Transgenic tobacco plants ectopically over-expressing PtGT1 were obtained and phenotypes were analysed. Wiesner and Mäule staining showed that stem xylem of transgenic tobacco plants stained more strongly than controls. Measurement of the Klason lignins showed much higher lignin content in the transgenic lines than in control plants. Furthermore, the ectopic over-expression of PtGT1 in tobacco resulted in an early flowering phenotype. These findings offer a possible starting point towards better understanding of the function of poplar PtGT1, and provide a novel strategy for lignin engineering and flowering control in plants through the genetic manipulation of a poplar glycosyltransferase gene. PMID:22268132

  12. RECOVERY - GEMINI-TITAN (GT)-IV - FROGMAN - ATLANTIC

    NASA Image and Video Library

    1965-06-07

    S65-33491 (7 June 1965) --- A United States Navy frogman team participates in the recovery of the Gemini-Titan 4 (GT-4) spacecraft. The USS Wasp was the prime recovery ship for the Gemini-4 mission. The crew of the Gemini-4 spaceflight was astronauts James A. McDivitt, command pilot, and Edward H. White II, pilot.

  13. The Development of the CryoTel™ LT and GT

    NASA Astrophysics Data System (ADS)

    Unger, Reuven

    2006-04-01

    This paper describes the considerations, design modifications and test results for the recently-developed Sunpower CryoTel™ LT and GT. After successful market introduction of the CryoTel™ CT, it became evident that some basic design modifications would render the unit available to a wider range of application. The CryoTel™ LT is a low-temperature variant of the original design. The LT's cooling capacity is 0.5W @ 23 K. The CryoTel™ GT is the enhanced performance variant. The GT's cooling capacity is 15 W @ 77 K. Both LT and GT largely retain the original structure and components and therefore benefit from the low-cost manufacturing profile of the original. Sunpower's main analysis and simulation tools were in-house codes and Gedeon Associates' SAGE Stirling cycle simulation. The CryoTel™ is a Linear Free Piston Integral Stirling cryocooler that makes use of Gas Bearing technology for non-contact operation and a microprocessor based driver/controller with a closed-loop temperature control.

  14. GEMINI-TITAN (GT)-4 - EARTH-SKY - OUTER SPACE

    NASA Image and Video Library

    1965-06-03

    S65-34776 (3-7 June 1965) --- This photograph shows the Nile Delta, Egypt, the Suez Canal, Israel, Jordan, Syria, Saudi Arabia, and Iraq as seen from the Gemini-Titan 4 (GT-4) spacecraft during its 12th revolution of Earth.

  15. [NRC/GT: Six Year One Research Studies.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean, Ed.

    1992-01-01

    This newsletter focuses on six Year 1 research projects associated with the National Research Center on the Gifted and Talented (NRC/GT). The updates address: "Regular Classroom Practices With Gifted Students: Findings from the Classroom Practices Survey" (Francis X. Archambault, Jr. and others); "The Classroom Practices Study:…

  16. PRELAUNCH - GEMINI-TITAN (GT)-3 - MISC. - CAPE

    NASA Image and Video Library

    1965-02-26

    S65-22666 (8 March 1965) --- Astronaut Walter M. Schirra Jr., the command pilot of the GT-3 backup crew, is shown suited up for prelaunch tests. He is shown with his helmet visor up and a thermometer in his mouth.

  17. Use of Conversion Adaptors to Clone Antigen Genes in Lambda gt11

    DTIC Science & Technology

    1987-01-01

    80*C for fu- estimated by comparison to molecular ture use. weight standards on ethidium bromide- DNA purification and manipulation. R. stained...restriction fragments suit- lated prior to ligation. High- molecular - ably sized (upper limit 8 kb) for insertion weight insert DNA was selectively precipi...epitopes identified in each library are listed in column 5 (antigen molecular mass in kDa). + Sau96I, Ddel, and Hinfl libraries are corn- fled sera were

  18. Helicobacter hepaticus Hh0072 gene encodes a novel alpha1-3-fucosyltransferase belonging to CAZy GT11 family.

    PubMed

    Zhang, Lei; Lau, Kam; Cheng, Jiansong; Yu, Hai; Li, Yanhong; Sugiarto, Go; Huang, Shengshu; Ding, Li; Thon, Vireak; Wang, Peng G; Chen, Xi

    2010-09-01

    Lewis x (Le(x)) and sialyl Lewis x (SLe(x))-containing glycans play important roles in numerous physiological and pathological processes. The key enzyme for the final step formation of these Lewis antigens is alpha1-3-fucosyltransferase. Here we report molecular cloning and functional expression of a novel Helicobacter hepaticus alpha1-3-fucosyltransferase (HhFT1) which shows activity towards both non-sialylated and sialylated Type II oligosaccharide acceptor substrates. It is a promising catalyst for enzymatic and chemoenzymatic synthesis of Le(x), sialyl Le(x) and their derivatives. Unlike all other alpha1-3/4-fucosyltransferases characterized so far which belong to Carbohydrate Active Enzyme (CAZy, http://www.cazy.org/) glycosyltransferase family GT10, the HhFT1 shares protein sequence homology with alpha1-2-fucosyltransferases and belongs to CAZy glycosyltransferase family GT11. The HhFT1 is thus the first alpha1-3-fucosyltransferase identified in the GT11 family.

  19. GT2RDF: Semantic Representation of Genetic Testing Data.

    PubMed

    Paul Rupa, Anamika; Singh, Sweta; Zhu, Qian

    2016-01-01

    Accelerated by the Human Genome Project, genetic testing has become an increasingly integral component in diagnosis, treatment, management, and prevention of numerous diseases and conditions. More than 480 laboratories perform genetic tests for more than 4,600 rare and common medical conditions. These tests can effectively help health professionals to determine or predict the genetic conditions of their patients. However, physicians have not actively incorporated such innovative genetic technology into their clinical practices according to two national wide surveys commissioned by UnitedHealth Group. To fill the gap of insufficient use of a large number of genetic tests, we generated a single Resource Description Framework (RDF) resource, called GT2RDF (Genetic Testing data to RDF) by integrating information about disease, gene, phenotype, genetic test, and drug from multiple sources including Genetic Testing Registry (GTR), Online Mendelian Inheritance in Man (OMIM), MedGen, Human Phenotype Ontology (HPO), ClinVar, National Drug File Reference Terminology (NDF-RT). Meanwhile, we manually annotated and extracted information from 200 randomly selected GeneReviews chapters, and integrated into the GT2RDF. We performed two case studies to demonstrate the usability of the GT2RDF. GT2RDF will serve as a data foundation to support the design of a genetic testing recommendation system, called iGenetics, which will ultimately facilitate the pace of precision medicine by means of actively and effectively incorporating innovative genetic technology in clinical settings. Abbreviations: GT2RDF: Genetic Testing data to RDF; SWT: Semantic web technology; OWL: Ontology Web Language; RDF: Resource Description Framework; SPARQL: SPARQL Protocol and RDF Query Language; GTR: Genetic Testing Registry; OMIM: Online Mendelian Inheritance in Man; HPO: Human Phenotype Ontology; NDF-RT: National Drug File Reference Terminology; UMLS: Unified Medical Language System.

  20. GT2RDF: Semantic Representation of Genetic Testing Data

    PubMed Central

    Paul Rupa, Anamika; Singh, Sweta; Zhu, Qian

    2016-01-01

    Accelerated by the Human Genome Project, genetic testing has become an increasingly integral component in diagnosis, treatment, management, and prevention of numerous diseases and conditions. More than 480 laboratories perform genetic tests for more than 4,600 rare and common medical conditions. These tests can effectively help health professionals to determine or predict the genetic conditions of their patients. However, physicians have not actively incorporated such innovative genetic technology into their clinical practices according to two national wide surveys commissioned by UnitedHealth Group. To fill the gap of insufficient use of a large number of genetic tests, we generated a single Resource Description Framework (RDF) resource, called GT2RDF (Genetic Testing data to RDF) by integrating information about disease, gene, phenotype, genetic test, and drug from multiple sources including Genetic Testing Registry (GTR), Online Mendelian Inheritance in Man (OMIM), MedGen, Human Phenotype Ontology (HPO), ClinVar, National Drug File Reference Terminology (NDF-RT). Meanwhile, we manually annotated and extracted information from 200 randomly selected GeneReviews chapters, and integrated into the GT2RDF. We performed two case studies to demonstrate the usability of the GT2RDF. GT2RDF will serve as a data foundation to support the design of a genetic testing recommendation system, called iGenetics, which will ultimately facilitate the pace of precision medicine by means of actively and effectively incorporating innovative genetic technology in clinical settings. Abbreviations: GT2RDF: Genetic Testing data to RDF; SWT: Semantic web technology; OWL: Ontology Web Language; RDF: Resource Description Framework; SPARQL: SPARQL Protocol and RDF Query Language; GTR: Genetic Testing Registry; OMIM: Online Mendelian Inheritance in Man; HPO: Human Phenotype Ontology; NDF-RT: National Drug File Reference Terminology; UMLS: Unified Medical Language System. PMID:28269903

  1. Why Clone?

    MedlinePlus

    ... disease. Find out more about Stem Cells . Reviving Endangered or Extinct Species You might have seen the Jurassic Park movies. ... related goat species to make a male. Cloning endangered species is much easier, mainly because the surviving animals ...

  2. GT-9 TEST - ASTRONAUT EDWARD H. WHITE -- MISCILANIES

    NASA Image and Video Library

    1965-06-03

    S65-19600 (3 June 1965) --- The prime crew for the Gemini-Titan 4 mission have an early morning breakfast prior to their historic flight which was launched at 10:16 a.m. (EST) on June 3, 1965. Shown here seated around the table (clockwise starting front center) are Dr. D. Owens Coons, chief, MSC Center Medical Office; astronaut James A. McDivitt, GT-4 command pilot; Dr. Eugene F. Tubbs, Kennedy Space Center; Rt. Rev. James Heiliky, McDivitt's priest at Cocoa Beach, Florida; Msgr. Irvine J. Nugent and astronaut Edward H. White II, GT-4 pilot. The group had a breakfast of tomato juice, broiled sirloin steak, poached eggs, toast, strawberry gelatin and coffee.

  3. Map-based cloning of the ALK gene, which controls the gelatinization temperature of rice.

    PubMed

    Gao, Zhenyu; Zeng, Dali; Cui, Xia; Zhou, Yihua; Yan, Meixian; Huang, Danian; Li, Jiayang; Qian, Qian

    2003-12-01

    Gelatinization temperature (GT) is an important parameter for evaluating the cooking and eating quality of rice besides amylose content (AC). The inheritance of the genes affecting GT has been widely studied and is considered to be controlled by a major gene. Here, we report the map-based cloning of rice ALK that encodes the soluble starch synthase II (SSSII). Comparison between the DNA sequences from different rice varieties, together with the results obtained with digestion of the rice seeds in alkali solution, indicates that the base substitutions in coding sequence of ALK may cause the alteration in GT.

  4. GT-CATS: Tracking Operator Activities in Complex Systems

    NASA Technical Reports Server (NTRS)

    Callantine, Todd J.; Mitchell, Christine M.; Palmer, Everett A.

    1999-01-01

    Human operators of complex dynamic systems can experience difficulties supervising advanced control automation. One remedy is to develop intelligent aiding systems that can provide operators with context-sensitive advice and reminders. The research reported herein proposes, implements, and evaluates a methodology for activity tracking, a form of intent inferencing that can supply the knowledge required for an intelligent aid by constructing and maintaining a representation of operator activities in real time. The methodology was implemented in the Georgia Tech Crew Activity Tracking System (GT-CATS), which predicts and interprets the actions performed by Boeing 757/767 pilots navigating using autopilot flight modes. This report first describes research on intent inferencing and complex modes of automation. It then provides a detailed description of the GT-CATS methodology, knowledge structures, and processing scheme. The results of an experimental evaluation using airline pilots are given. The results show that GT-CATS was effective in predicting and interpreting pilot actions in real time.

  5. EXTRAVEHICULAR ACTIVITY (EVA) - GEMINI-TITAN (GT)-4

    NASA Image and Video Library

    1965-06-03

    S65-29766 (3 June 1965) --- Astronaut Edward H. White II, pilot for the Gemini-Titan 4 (GT-4) spaceflight, floats in the zero-gravity of space during the third revolution of the GT-4 spacecraft. White wears a specially designed spacesuit. His face is shaded by a gold-plated visor to protect him from unfiltered rays of the sun. In his right hand he carries a Hand-Held Self-Maneuvering Unit (HHSMU) that gives him control over his movements in space. White also wears an emergency oxygen chest pack; and he carries a camera mounted on the HHSMU for taking pictures of the sky, Earth and the GT-4 spacecraft. He is secured to the spacecraft by a 25-feet umbilical line and a 23-feet tether line. Both lines are wrapped together in gold tape to form one cord. Astronaut James A. McDivitt, command pilot, remained inside the spacecraft during the extravehicular activity (EVA). Photo credit: NASA EDITOR'S NOTE: Astronaut Edward H. White II died in the Apollo/Saturn 204 fire at Cape Kennedy on Jan. 27, 1967.

  6. Molecular cloning.

    PubMed

    Lessard, Juliane C

    2013-01-01

    This protocol describes the basic steps involved in conventional plasmid-based cloning. The goals are to insert a DNA fragment of interest into a receiving vector plasmid, transform the plasmid into E. coli, recover the plasmid DNA, and check for correct insertion events.

  7. Gemini-Titan (GT)-3 - Prelaunch Activities - Cape

    NASA Image and Video Library

    1965-03-23

    S65-21093 (23 March 1965) --- Astronaut Virgil I. Grissom (facing camera at right), command pilot of the Gemini-Titan 3 flight, is shown during a steak breakfast which he was served about two hours prior to the 9:24 a.m. (EST) GT-3 launch on March 23, 1965. Pictured in the foreground are Donald K. Slayton (right), assistant director for Flight Crew Operations; and Walter Burke, general manager of McDonnell Aircraft Corporation Spacecraft and Missiles. Pictured in the background are astronaut Alan B. Shepard Jr. (left) and Walter C. Williams, former deputy director of the Manned Spacecraft Center, now with a private aerospace firm.

  8. GEMINI-TITAN (GT)-10 - EARTH SKY - RENDEZVOUS - OUTER SPACE

    NASA Image and Video Library

    1966-07-18

    S66-46122 (18 July 1966) --- Agena Target Docking Vehicle 5005 is photographed from the Gemini-Titan 10 (GT-10) spacecraft during rendezvous in space. The two spacecraft are about 38 feet apart. After docking with the Agena, astronauts John W. Young, command pilot, and Michael Collins, pilot, fired the 16,000 pound thrust engine of Agena X's primary propulsion system to boost the combined vehicles into an orbit with an apogee of 413 nautical miles to set a new altitude record for manned spaceflight. Photo credit: NASA

  9. Little phenotypic variability in three CF sibs compound heterozygous for the 621 + 1G-->T and the 711 + 1G-->T mutations.

    PubMed

    De Braekeleer, M; Simard, F; Aubin, G

    1997-03-01

    We describe a family in which three sibs are compound heterozygotes for two rather rare CFTR splice-site mutations, the 621 + 1G-->T and the 711 + 1G-->T mutations. Little phenotypic variation was observed between sibs, of whom two are deceased. Their disease is characterized by pancreatic insufficiency, a severe pulmonary involvement and major growth retardation.

  10. Turbulent transport measurements in a model of GT-combustor

    NASA Astrophysics Data System (ADS)

    Chikishev, L. M.; Gobyzov, O. A.; Sharaborin, D. K.; Lobasov, A. S.; Dulin, V. M.; Markovich, D. M.; Tsatiashvili, V. V.

    2016-10-01

    To reduce NOx formation modern industrial power gas-turbines utilizes lean premixed combustion of natural gas. The uniform distribution of local fuel/air ratio in the combustion chamber plays one of the key roles in the field of lean combustion to prevent thermo-acoustic pulsations. Present paper reports on simultaneous Particle Image Velocimetry and acetone Planar Laser Induced Fluorescence measurements in a cold model of GT-combustor to investigate mixing processes which are relevant to the organization of lean premixed combustion. Velocity and passive admixture pulsations correlations were measured to verify gradient closer model, which is often used in Reynolds-Averaged Navier-Stokes (RANS) simulation of turbulent mixing.

  11. ROUNDUP - EMBLEM - GEMINI-TITAN (GT)-11 - MSC

    NASA Image and Video Library

    1966-09-01

    S66-44308 (September 1966) --- Insignia of the Gemini-Titan XI (GT-11) spaceflight. Roman numeral indicates eleventh flight in the Gemini series. Two spacecraft symbolize rendezvous and docking of Gemini with an Agena. Astronaut and umbilical (tether) line denotes planned extravehicular activity. Astronauts Charles Conrad Jr., command pilot, and Richard F. Gordon Jr., pilot, are members of the Gemini-11 prime crew. The NASA insignia design for Gemini flights is reserved for use by the astronauts and for other official use as the NASA Administrator may authorize. Public availability has been approved only in the form of illustrations by the various news media. When and if there is any change in this policy, which we do not anticipate, it will be publicly announced. Photo credit: NASA

  12. WBP: The wood Brazilian BIG-GT demonstration project

    SciTech Connect

    Carpentieri, E.

    1993-12-31

    Brazil is one of the leading countries in the use of renewable energy. Most of its electricity comes from hydro power, about 200,000 barrels a day of ethanol from sugar cane is used as fuel, around 38% of the pig iron, and 20% of the steel production, uses charcoal as a reducing medium. Located in the tropics, with the sun shining all year round, and with its vast territory, the Country may be regarded as having all the basic conditions to develop a modern Biomass for Electricity industry. The conjunction of those characteristics with, the necessity of developing new energy resources for electricity production in the Northeast of the Country, the results of the studies made by Princeton University, Shell and Chesf, the progress achieved by the BIG-GT (Biomass Integrated Gasification Gas Turbine) technology in Europe, and the organization of the Global Environment Facility (GEF), provided the unique opportunity for the implementation of a commercial demonstration in Brazil. This paper describes the idea, the scope, the technical challenges, and actual status of development of the WBP, a project which aims to demonstrate the commercial viability of the BIG-GT technology. It also highlights, the project management structure, the role of the GEF, World Bank and of the United Nations Development Program (UNDP), and the participation of the Brazilian Federal Government, through the Ministry of Science and Technology (MCT). Finally it describes the Participants (ELETROBRAS, CVRD, CIENTEC, SHELL, and CHESF), their role in the project, and how the group was formed and operates.

  13. [Eugenics and human cloning].

    PubMed

    Boloz, W

    2001-01-01

    Because of legislative bans there are still no reports of human cloning. However eager public debate is currently running, concerning medical, legal, social and ethical aspects of human cloning. Arguments for and against human cloning are presented. An important argument against cloning is the danger of eugenic tendencies connected with cloning, which could lead to genetic discrimination.

  14. 46 CFR 125.125 - Carriage of noxious liquid substances in bulk by OSVs of at least 6,000 GT ITC (500 GRT if GT ITC...

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... reference, see § 125.180), except that cargoes with an “S” designation in the hazard column (column d) in... (500 GRT if GT ITC is not assigned) that meets the stability and cargo tank location requirements— (1...

  15. Malignant pericytes expressing GT198 give rise to tumor cells through angiogenesis

    PubMed Central

    Zhang, Liyong; Wang, Yan; Rashid, Mohammad H.; Liu, Min; Angara, Kartik; Mivechi, Nahid F.; Maihle, Nita J.; Arbab, Ali S.; Ko, Lan

    2017-01-01

    Angiogenesis promotes tumor development. Understanding the crucial factors regulating tumor angiogenesis may reveal new therapeutic targets. Human GT198 (PSMC3IP or Hop2) is an oncoprotein encoded by a DNA repair gene that is overexpressed in tumor stromal vasculature to stimulate the expression of angiogenic factors. Here we show that pericytes expressing GT198 give rise to tumor cells through angiogenesis. GT198+ pericytes and perivascular cells are commonly present in the stromal compartment of various human solid tumors and rodent xenograft tumor models. In human oral cancer, GT198+ pericytes proliferate into GT198+ tumor cells, which migrate into lymph nodes. Increased GT198 expression is associated with increased lymph node metastasis and decreased progression-free survival in oral cancer patients. In rat brain U-251 glioblastoma xenografts, GT198+ pericytes of human tumor origin encase endothelial cells of rat origin to form mosaic angiogenic blood vessels, and differentiate into pericyte-derived tumor cells. The net effect is continued production of glioblastoma tumor cells from malignant pericytes via angiogenesis. In addition, activation of GT198 induces the expression of VEGF and promotes tube formation in cultured U251 cells. Furthermore, vaccination using GT198 protein as an antigen in mouse xenograft of GL261 glioma delayed tumor growth and prolonged mouse survival. Together, these findings suggest that GT198-expressing malignant pericytes can give rise to tumor cells through angiogenesis, and serve as a potential source of cells for distant metastasis. Hence, the oncoprotein GT198 has the potential to be a new target in anti-angiogenic therapies in human cancer. PMID:28881671

  16. The National Research Center on the Gifted and Talented (NRC/GT) Newsletter, 1999.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean, Ed.; Siegle, Del, Ed.

    1999-01-01

    These two newsletters of The National Research Center on the Gifted and Talented (NRC/GT) present articles concerned with research on the education of gifted and talented students. The articles include: "NRC/GT: Making Decisions and Determining Next Steps" (E. Jean Gubbins); "Free Summer Programs for Talented Teens" (D. Betsy McCoach); "High End…

  17. Developing the Gifts and Talents of All America's Students: NRC/GT--1990-1995.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean; And Others

    This monograph discusses the organization and mission of the National Research Center on the Gifted and Talented (NRC/GT) and describes the research studies and commissioned papers that the Center has sponsored. Part 1, "Dream and Design for the NRC/GT," provides an overview of the Center as an organization and describes how the research…

  18. The National Research Center on the Gifted and Talented (NRC/GT) Newsletter, 1998.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean, Ed.; Siegle, Del, Ed.

    1998-01-01

    These two newsletters of The National Research Center on the Gifted and Talented (NRC/GT) present articles concerned with research on the education of gifted and talented students. The articles are: "NRC/GT's Suggestions: Evaluating Your Programs and Services" (E. Jean Gubbins); "Professional Development Practices in Gifted Education: Results of a…

  19. DuoliteTM GT-73 Resin Testing in Support of the Salt Disposition Alternatives

    SciTech Connect

    Wilmarth, W.R.

    1998-12-07

    This study evaluated DuoliteTM GT-73 performance for removing mercury ions from several high level waste streams under consideration by the Salt Disposition Systems Engineering Team as a technical risk. Experiments conducted over an eight week period address the technical uncertainties for GT-73 performance

  20. Complete Genome Sequence of Bifidobacterium longum GT15: Unique Genes for Russian Strains

    PubMed Central

    Zakharevich, Natalia V.; Averina, Olga V.; Klimina, Ksenia M.; Kudryavtseva, Anna V.; Kasianov, Artem S.; Makeev, Vsevolod J.

    2014-01-01

    In this study, we report the first completely annotated genome sequence of the Russian-origin Bifidobacterium longum subsp. longum strain GT15. We discovered 35 unique genes (UGs) which were detected from only the B. longum GT15 genome and were absent from other B. longum strain genomes (not of Russian origin). PMID:25523785

  1. Helium turbomachine design for GT-MHR power plant

    SciTech Connect

    McDonald, C.F.; Orlando, R.J.; Cotzas, G.M.

    1994-07-01

    The power conversion system in the gas turbine modular helium reactor (GT-MHR) power plant is based on a highly recuperated closed Brayton cycle. The major component in the direct cycle system is a helium closed-cycle gas turbine rated at 286 MW(e). The rotating group consists of an intercooled helium turbocompressor coupled to a synchronous generator. The vertical rotating assembly is installed in a steel vessel, together with the other major components (i.e., recuperator, precooler, intercooler, and connecting ducts and support structures). The rotor is supported on an active magnetic bearing system. The turbine operates directly on the reactor helium coolant, and with a temperature of 850{degree}C (1562{degree}F) the plant efficiency is over 47%. This paper addresses the design and development planning of the helium turbomachine, and emphasizes that with the utilization of proven technology, this second generation nuclear power plant could be in service in the first decade of the 21st century.

  2. A photometric and orbital analysis of GT MUSCAE

    NASA Astrophysics Data System (ADS)

    Murdoch, K. A.; Hearnshaw, J. B.; Kilmartin, P. M.; Gilmore, A. C.

    1995-10-01

    GT Mus is a quadruple system comprising a long-period RS CVn-type binary (HD 101379) and a pair of eclipsing A dwarfs (HD 101380). Six and a half years of UBV (RI)_C photometry obtained at the Mt John University Observatory has enabled identification of four distinct types of photometric variability in this system. These are (1) a slowly changing mean magnitude, which probably arises from an activity-cycle-like effect in the active component of HD 101379, (2) a periodic variation (P_rot~64d), which is attributed to rotational modulation due to spots on the active star, (3) a periodic variation (P_eclipse=2.7546d) due to the eclipses of HD 101380, and (4) an excess in the I band, which occurs on a short time-scale (<1d) and is probably associated with HD 101379 activity. The evolution of the light curve of HD 101379 is fast with respect to the rotational period, suggesting rapid spot evolution for which we anticipate a possible model. The colours of HD 101379, even at maximum brightness, are excessively red for its spectral type, unless there is significant reddening by dust. Radial velocity measurements of HD 101379 are also presented, along with an improved determination of the orbit of this somewhat long-period (P_orb=61.448d) system.

  3. The GT-MHR for destruction of weapons plutonium

    SciTech Connect

    Baxter, A.M.; Neylan, A.J.

    1995-12-31

    The disposal of nearly 100 tonnes of weapons-grade plutonium (WG-Pu) made surplus by the disarmament treaties is receiving urgent attention, highlighted by the recent seizure in Germany of small quantities of weapons-useful plutonium. Unlike highly enriched uranium, simple denaturing cannot make this plutonium worthless for use in future weapons. The use of physical security and institutional barriers, including long-term storage in high-level waste repositories, to provide secure storage for centuries to come is questionable when considering government instability and the possibility of national recidivism. The Russian Ministry for Atomic Energy (MINATOM) and General Atomics have signed an agreement for the cooperative design of a gas turbine-modular helium reactor (GT-MHR) to burn the WG-Pu stockpile. A formal proposal for a joint U.S./Russian program for the development of this reactor has been submitted by MINATOM to Vice President Gore. The major benefit of this program is that the reactor would deplete the Russian surplus plutonium stockpile, provide jobs for technical specialists in the former weapons complex, and produce valuable electric power. It would also provide a mutually assured means of destroying the U.S. and Russian stockpiles.

  4. On classical cloning and no-cloning

    NASA Astrophysics Data System (ADS)

    Teh, Nicholas J.

    2012-02-01

    It is part of information theory folklore that, while quantum theory prohibits the generic (or universal) cloning of states, such cloning is allowed by classical information theory. Indeed, many take the phenomenon of no-cloning to be one of the features that distinguishes quantum mechanics from classical mechanics. In this paper, we argue that pace conventional wisdom, in the case where one does not include a machine system, there is an analog of the no-cloning theorem for classical systems. However, upon adjoining a non-trivial machine system (or ancilla) one finds that, pace the quantum case, the obstruction to cloning disappears for pure states. We begin by discussing some conceptual points and category-theoretic generalities having to do with cloning, and proceed to discuss no-cloning in both the case of (non-statistical) classical mechanics and classical statistical mechanics.

  5. Molecular cloning and amino acid sequence of human 5-lipoxygenase

    SciTech Connect

    Matsumoto, T.; Funk, C.D.; Radmark, O.; Hoeoeg, J.O.; Joernvall, H.; Samuelsson, B.

    1988-01-01

    5-Lipoxygenase (EC 1.13.11.34), a Ca/sup 2 +/- and ATP-requiring enzyme, catalyzes the first two steps in the biosynthesis of the peptidoleukotrienes and the chemotactic factor leukotriene B/sub 4/. A cDNA clone corresponding to 5-lipoxygenase was isolated from a human lung lambda gt11 expression library by immunoscreening with a polyclonal antibody. Additional clones from a human placenta lambda gt11 cDNA library were obtained by plaque hybridization with the /sup 32/P-labeled lung cDNA clone. Sequence data obtained from several overlapping clones indicate that the composite DNAs contain the complete coding region for the enzyme. From the deduced primary structure, 5-lipoxygenase encodes a 673 amino acid protein with a calculated molecular weight of 77,839. Direct analysis of the native protein and its proteolytic fragments confirmed the deduced composition, the amino-terminal amino acid sequence, and the structure of many internal segments. 5-Lipoxygenase has no apparent sequence homology with leukotriene A/sub 4/ hydrolase or Ca/sup 2 +/-binding proteins. RNA blot analysis indicated substantial amounts of an mRNA species of approx. = 2700 nucleotides in leukocytes, lung, and placenta.

  6. The Clone Factory

    ERIC Educational Resources Information Center

    Stoddard, Beryl

    2005-01-01

    Have humans been cloned? Is it possible? Immediate interest is sparked when students are asked these questions. In response to their curiosity, the clone factory activity was developed to help them understand the process of cloning. In this activity, students reenact the cloning process, in a very simplified simulation. After completing the…

  7. The Clone Factory

    ERIC Educational Resources Information Center

    Stoddard, Beryl

    2005-01-01

    Have humans been cloned? Is it possible? Immediate interest is sparked when students are asked these questions. In response to their curiosity, the clone factory activity was developed to help them understand the process of cloning. In this activity, students reenact the cloning process, in a very simplified simulation. After completing the…

  8. Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola.

    PubMed

    Taverniers, Isabel; Windels, Pieter; Vaïtilingom, Marc; Milcamps, Anne; Van Bockstaele, Erik; Van den Eede, Guy; De Loose, Marc

    2005-04-20

    Since the 18th of April 2004, two new regulations, EC/1829/2003 on genetically modified food and feed products and EC/1830/2003 on traceability and labeling of GMOs, are in force in the EU. This new, comprehensive regulatory framework emphasizes the need of an adequate tracing system. Unique identifiers, such as the transgene genome junction region or a specific rearrangement within the transgene DNA, should form the basis of such a tracing system. In this study, we describe the development of event-specific tracing systems for transgenic maize lines Bt11, Bt176, and GA21 and for canola event GT73. Molecular characterization of the transgene loci enabled us to clone an event-specific sequence into a plasmid vector, to be used as a marker, and to develop line-specific primers. Primer specificity was tested through qualitative PCRs and dissociation curve analysis in SYBR Green I real-time PCRs. The primers were then combined with event-specific TaqMan probes in quantitative real-time PCRs. Calibration curves were set up both with genomic DNA samples and the newly synthesized plasmid DNA markers. It is shown that cloned plasmid GMO target sequences are perfectly suitable as unique identifiers and quantitative calibrators. Together with an event-specific primer pair and a highly specific TaqMan probe, the plasmid markers form crucial components of a unique and straighforward tracing system for Bt11, Bt176, and GA21 maize and GT73 canola events.

  9. GT-2: in vivo transcriptional activation activity and definition of novel twin DNA binding domains with reciprocal target sequence selectivity.

    PubMed

    Ni, M; Dehesh, K; Tepperman, J M; Quail, P H

    1996-06-01

    GT-2 is a novel DNA binding protein that interacts with a triplet functionally defined, positively acting GT-box motifs (GT1-bx, GT2-bx, and GT3-bx) in the rice phytochrome A gene (PHYA) promoter. Data from a transient transfection assay used here show that recombinant GT-2 enhanced transcription from both homologous and heterologous GT-box-containing promoters, thereby indicating that this protein can function as a transcriptional activator in vivo. Previously, we have shown that GT-2 contains separate DNA binding determinants in its N- and C-terminal halves, with binding site preferences for the GT3-bx and GT2-bx promoter motifs, respectively. Here, we demonstrate that the minimal DNA binding domains reside within dual 90-amino acid polypeptide segments encompassing duplicated sequences, termed trihelix regions, in each half of the molecule, plus 15 additional immediately adjacent amino acids downstream. These minimal binding domains retained considerable target sequence selectivity for the different GT-box motifs, but this selectivity was enhanced by a separate polypeptide segment farther downstream on the C-terminal side of each trihelix region. Therefore, the data indicate that the twin DNA binding domains of GT-2 each consist of a general GT-box recognition core with intrinsic differential binding activity toward closely related target motifs and a modified sequence conferring higher resolution reciprocal selectivity between these motifs.

  10. Analysis of CFTR transcripts in nasal epithelial cells and lymphoblasts of a cystic fibrosis patient with 621 + 1G-->T and 711 + 1G-->T mutations.

    PubMed

    Zielenski, J; Bozon, D; Markiewicz, D; Aubin, G; Simard, F; Rommens, J M; Tsui, L C

    1993-06-01

    We have analyzed the CFTR mRNA populations in a cystic fibrosis patient heterozygous for the 621 + 1G-->T and 711 + 1G-->T mutations. Total RNA isolated from the nasal epithelial cells and Epstein-Barr virus-transformed lymphoblasts derived from this patient was reversely transcribed and a region extending from exon 3 to exon 7 of the gene was amplified by the polymerase chain reaction and analyzed. Three abnormal products were identified, suggesting the presence of three aberrant transcripts, and their profiles were identical in both cell types. Two of the products were found to be missing either exon 4 or exon 5 as anticipated from the transcripts from the 621 + 1G-->T or 711 + 1G-->T alleles, respectively. The third product was apparently derived from an alternatively spliced mRNA species in the absence of the nominal splice site (in 621 + 1G-->T) through the use of a cryptic splice donor sequence (TT528/GTGAGG) within exon 4. Although reading frames appeared to be preserved in all three putative transcripts, significant portions of the presumed first and second transmembrane spans as well as the immediately following cytoplasmic domain would be deleted from the mutant CFTR polypeptides, if made. These observations are consistent with a loss of CFTR function in this cystic fibrosis patient.

  11. GT160-246, a toxin binding polymer for treatment of Clostridium difficile colitis.

    PubMed

    Kurtz, C B; Cannon, E P; Brezzani, A; Pitruzzello, M; Dinardo, C; Rinard, E; Acheson, D W; Fitzpatrick, R; Kelly, P; Shackett, K; Papoulis, A T; Goddard, P J; Barker, R H; Palace, G P; Klinger, J D

    2001-08-01

    GT160-246, a high-molecular-weight soluble anionic polymer, was tested in vitro and in vivo for neutralization of Clostridium difficile toxin A and B activities. Five milligrams of GT160-246 per ml neutralized toxin-mediated inhibition of protein synthesis in Vero cells induced by 5 ng of toxin A per ml or 1.25 ng of toxin B per ml. In ligated rat ileal loops, 1 mg of GT160-246 neutralized fluid accumulation caused by 5 microg of toxin A. At doses as high as 80 mg/loop, cholestyramine provided incomplete neutralization of fluid accumulation caused by 5 microg of toxin A. GT160-246 protected 80% of the hamsters from mortality caused by infection with C. difficile, whereas cholestyramine protected only 10% of animals. Treatment of C. difficile-infected hamsters with metronidazole initially protected 100% of the hamsters from mortality, but upon removal of treatment, 80% of the hamsters had relapses and died. In contrast, removal of GT160-246 treatment did not result in disease relapse in the hamsters. GT160-246 showed no antimicrobial activity in tests with a panel of 16 aerobic bacteria and yeast and 22 anaerobic bacteria and did not interfere with the in vitro activities of most antibiotics. GT160-246 offers a novel, nonantimicrobial treatment of C. difficile disease in humans.

  12. Multi-color lightcurve observation of the asteroid (163249) 2002 GT

    NASA Astrophysics Data System (ADS)

    Oshima, M.; Abe, S.

    2014-07-01

    NASA's Deep Impact/EPOXI spacecraft plans to encounter the asteroid (163249) 2002 GT, classified as a PHA (Potentially Hazardous Asteroid), on January 4, 2020. However, the taxonomic type and spin state of 2002 GT remain to be determined. We have carried out ground-based multi-color (B-V-R-I) lightcurve observations taking advantage of the 2002 GT Characterization Campaign by NASA. Multi-color lightcurve measurements allow us to estimate the rotation period and obtain strong constraints on the shape and pole orientation. Here we found that the rotation period of 2002 GT is estimated to be 3.7248 ± 0.1664 h. In mid-2013, 2002 GT passed at 0.015 au from the Earth, resulting an exceptional opportunity for ground-based characterization. Using the 0.81-m telescope of the Tenagra Observatory (110°52'44.8''W, +31°27'44.4''N, 1312 m) in Arizona, USA, and the Johnson-Cousins BVRI filters, we have found lightcurves of 2002 GT (Figure). The Tenagra II 0.81-m telescope is used for research of the Hayabusa2 target Asteroid (162173) 1999 JU_3. The lightcurves (relative magnitude) show that the rotation period of 2002 GT, the target of NASA's Deep Impact/EPOXI spacecraft, is estimated to be 3.7248 ± 0.1664 hr. On June 9, 2013, we had 7 hours of ground-based observations on 2002 GT from 4:00 to 11:00 UTC. The number of comparison stars for differential photometry was 34. Because of tracking the fast-moving asteroid, it was necessary to have the same comparison star among the fields of vision. We have also obtained absolute photometry of 2002 GT on June 13, 2013.

  13. Development of Orally Administered γ-Tocotrienol (GT3) Nanoemulsion for Radioprotection

    PubMed Central

    Ledet, Grace A.; Biswas, Shukla; Kumar, Vidya P.; Graves, Richard A.; Mitchner, Demaurian M.; Parker, Taylor M.; Bostanian, Levon A.; Ghosh, Sanchita P.; Mandal, Tarun K.

    2016-01-01

    The purpose of this study was two-fold: (1) to formulate γ-tocotrienol (GT3) in a nanoemulsion formulation as a prophylactic orally administered radioprotective agent; and (2) to optimize the storage conditions to preserve the structural integrity of both the formulation and the compound. γ-tocotrienol was incorporated into a nanoemulsion and lyophilized with lactose. Ultra performance liquid chromatography–mass spectroscopy (UPLC–MS) was used to monitor the chemical stability of GT3 over time, the particle size and ζ potential, and scanning electron microscopy (SEM) were used to study the physical stability of the nanoemulsion. Radioprotective and toxicity studies were performed in mice. The liquid formulation exhibited GT3 degradation at all storage temperatures. Lyophilization, in the presence of lactose, significantly reduced GT3 degradation. Both the liquid and lyophilized nanoemulsions had stable particle size and ζ potential when stored at 4 °C. Toxicity studies of the nanoemulsion resulted in no observable toxicity in mice at an oral dose of 600 mg/kg GT3. The nano-formulated GT3 (300 mg/kg) demonstrated enhanced survival efficacy compared to GT3 alone (200 and 400 mg/kg) in CD2F1 mice exposed to total body gamma radiation. The optimal long-term storage of formulated GT3 is as a powder at −20 °C to preserve drug and formulation integrity. Formulation of GT3 as a nanoemulsion for oral delivery as a prophylactic radioprotectant shows promise and warrants further investigation. PMID:28029115

  14. Development of Orally Administered γ-Tocotrienol (GT3) Nanoemulsion for Radioprotection.

    PubMed

    Ledet, Grace A; Biswas, Shukla; Kumar, Vidya P; Graves, Richard A; Mitchner, Demaurian M; Parker, Taylor M; Bostanian, Levon A; Ghosh, Sanchita P; Mandal, Tarun K

    2016-12-24

    The purpose of this study was two-fold: (1) to formulate γ-tocotrienol (GT3) in a nanoemulsion formulation as a prophylactic orally administered radioprotective agent; and (2) to optimize the storage conditions to preserve the structural integrity of both the formulation and the compound. γ-tocotrienol was incorporated into a nanoemulsion and lyophilized with lactose. Ultra performance liquid chromatography-mass spectroscopy (UPLC-MS) was used to monitor the chemical stability of GT3 over time, the particle size and ζ potential, and scanning electron microscopy (SEM) were used to study the physical stability of the nanoemulsion. Radioprotective and toxicity studies were performed in mice. The liquid formulation exhibited GT3 degradation at all storage temperatures. Lyophilization, in the presence of lactose, significantly reduced GT3 degradation. Both the liquid and lyophilized nanoemulsions had stable particle size and ζ potential when stored at 4 °C. Toxicity studies of the nanoemulsion resulted in no observable toxicity in mice at an oral dose of 600 mg/kg GT3. The nano-formulated GT3 (300 mg/kg) demonstrated enhanced survival efficacy compared to GT3 alone (200 and 400 mg/kg) in CD2F1 mice exposed to total body gamma radiation. The optimal long-term storage of formulated GT3 is as a powder at -20 °C to preserve drug and formulation integrity. Formulation of GT3 as a nanoemulsion for oral delivery as a prophylactic radioprotectant shows promise and warrants further investigation.

  15. Dynamics and control modeling of the closed-cycle gas turbine (GT-HTGR) power plant

    SciTech Connect

    Bardia, A.

    1980-02-01

    The simulation if presented for the 800-MW(e) two-loop GT-HTGR plant design with the REALY2 transient analysis computer code, and the modeling of control strategies called for by the inherently unique operational requirements of a multiple loop GT-HTGR is described. Plant control of the GT-HTGR is constrained by the nature of its power conversion loops (PCLs) in which the core cooling flow and the turbine flow are directly related and thus changes in flow affect core cooling as well as turbine power. Additionally, the high thermal inertia of the reactor core precludes rapid changes in the temperature of the turbine inlet flow.

  16. Molecular cloning and biochemical characterization of the UDP-glucose: flavonoid 3-O-glucosyltransferase from Concord grape (Vitis labrusca).

    PubMed

    Hall, Dawn; Yuan, Xiao Xin; Murata, Jun; De Luca, Vincenzo

    2012-02-01

    Glucosylation of anthocyanidin substrates at the 3-O-position is crucial for the red pigmentation of grape berries and wine. The gene that encodes the enzyme involved in this reaction has been cloned from Vitis labrusca cv. Concord, heterologously expressed, and the recombinant enzyme (rVL3GT) was characterized. VL3GT has 96% amino acid sequence identity with Vitis vinifera VV3GT and groups phylogenetically with several other flavonoid 3-O-glycosyltransferases. In vitro substrate specificity studies and kinetic analyses of rVL3GT indicate that this enzyme preferentially glucosylates cyanidin as compared with quercetin. Crude protein extracts from several Concord grape tissues were assayed for glucosyltransferase activity with cyanidin and quercetin as acceptor substrates. A comparison of the VL3GT activities toward with these substrates showed that the 3GT enzyme activity is consistent with the expression of VL3GT in these tissues and is coincident with the biosynthesis of anthocyanins in both location and developmental stages. Enzyme activities in grape mesocarp, pre-veraison exocarp, leaf, flower bud, and flower tissues glucosylated quercetin but not cyanidin at high rates, suggesting the presence of additional enzymes which are able to glucosylate the 3-O-position of flavonols with higher specificity than anthocyanidins. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. The National Research Center on the Gifted and Talented (NRC/GT) Newsletter. Fall 1994-Spring 1995.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean, Ed.; Siegle, Del, Ed.

    1995-01-01

    This document consists of three consecutive but unnumbered issues of a newsletter from the National Research Center on the Gifted and Talended (NRC/GT) containing articles on the education of gifted and talented students: "NRC/GT Destination: Around the Corner" (E. Jean Gubbins); "New NRC/GT Studies for Year 5" (on implementing enrichment…

  18. Identification of cDNA clones expressing immunodiagnostic antigens from Trichinella spiralis

    SciTech Connect

    Zarlenga, D.; Gamble, H.R.

    1987-05-01

    A cDNA expression library was built in lambda gt11 phage using poly A mRNA isolated from Trichinella spiralis muscle stage larvae. This library was screened with rabbit antibodies to parasite excretory-secretory (ES) products and greater than 180 clones were isolated. Thirteen clones producing highly immunogenic protein antigens were plaque purified and rescreened with pig antisera to T.spiralis, Trichuris suis or Ascaris suum to identify clones producing epitopes specific to T.spiralis ES products, only. Two clones, TsAc-2 and TsAc-8, which displayed strong interactions with pig antisera to T. spiralis were lysogenized in E. coli Y1089 and the protein extracted. Western blots of the crude fusion proteins revealed molecular weights of 133 kD and 129 kD, respectively. Northern blot analysis of total RNA with TSP labelled cDNA:lambda gt11 probes indicated single RNA transcripts for each clone with molecular sizes corresponding to 800-850 nucleotides. dscDNA inserts were estimated by southern blot analysis to be 500 bp and 340 bp, respectively, with no cross-hybridization observed between the cloned sequences. Dot blots using pig sera to screen crude fusion protein preparations, total bacterial protein (negative controls) and crude worm extract or ES products from T.spiralis, T.suis and A.suum (positive controls) corroborated the specificity and sensitivity of these clones as potential diagnostic antigens for swine trichinellosis.

  19. Energy recuperation in solid oxide fuel cell (SOFC) and gas turbine (GT) combined system

    NASA Astrophysics Data System (ADS)

    Kuchonthara, Prapan; Bhattacharya, Sankar; Tsutsumi, Atsushi

    A combined power generation system consisting of a solid oxide fuel cell (SOFC) and a gas turbine (GT) with steam and heat recuperation (HR) was evaluated using a commercial process simulation tool, ASPEN Plus. The effect of steam recuperation (SR) on the overall efficiency of the combined system was investigated by comparing the SOFC-GT during heat and steam recuperation (HSR) against the system during only heat recuperation. At low turbine inlet temperatures (TITs), the overall efficiency of the SOFC-GT combined system with heat and steam recuperation improved by showing an increase in TIT and a reduction in pressure ratio (PR). On the other hand, at high TITs, the opposite trend was observed. The integration of steam recuperation was found to improve the overall efficiency and specific power of SOFC-GT combined systems with a relatively compact SOFC component.

  20. v-Src activates the expression of 92-kDa type IV collagenase gene through the AP-1 site and the GT box homologous to retinoblastoma control elements. A mechanism regulating gene expression independent of that by inflammatory cytokines.

    PubMed

    Sato, H; Kita, M; Seiki, M

    1993-11-05

    The 92-kDa type IV collagenase (matrix metalloproteinase-9; MMP-9) is frequently expressed in cells showing an invasive nature during physiological and pathological processes, and the expression is strictly controlled by a variety of trans-membrane signals. Binding sites for NF-kB, Sp-1, and AP-1 are reportedly required for induction of MMP-9 gene expression by tumor necrosis factor-alpha or 12-O-tetradecanoylphorbol-13-acetate. Comparison of the sequence of the newly cloned mouse MMP-9 promoter region with our previous human isolate revealed that, in addition to the above mentioned elements, four units of GGGG(T/A)GGGG sequence (GT box) were conserved between the two species. In this study, we have demonstrated that one of the GT boxes located downstream of the AP-1 site is essential along with the AP-1 site for the activation of the promoter by v-Src but not by tumor necrosis factor-alpha or 12-O-tetradecanoylphorbol-13-acetate. Gel mobility-shift assays revealed that binding proteins for retinoblastoma control element, including Sp-1 family protein, can bind specifically to GT boxes. Thus, the v-Src signals to the AP-1 site and to the GT box homologous to retinoblastoma control element acted synergistically in transcriptional activation. These results suggest that certain v-Src-mediated signals are propagated along pathways that are independent of inflammatory cytokines.

  1. Multipartite asymmetric quantum cloning

    SciTech Connect

    Iblisdir, S.; Gisin, N.; Acin, A.; Cerf, N.J.; Filip, R.; Fiurasek, J.

    2005-10-15

    We investigate the optimal distribution of quantum information over multipartite systems in asymmetric settings. We introduce cloning transformations that take N identical replicas of a pure state in any dimension as input and yield a collection of clones with nonidentical fidelities. As an example, if the clones are partitioned into a set of M{sub A} clones with fidelity F{sup A} and another set of M{sub B} clones with fidelity F{sup B}, the trade-off between these fidelities is analyzed, and particular cases of optimal N{yields}M{sub A}+M{sub B} cloning machines are exhibited. We also present an optimal 1{yields}1+1+1 cloning machine, which is an example of a tripartite fully asymmetric cloner. Finally, it is shown how these cloning machines can be optically realized.

  2. Aristotle and headless clones.

    PubMed

    Mosteller, Timothy

    2005-01-01

    Cloned organisms can be genetically altered so that they do not exhibit higher brain functioning. This form of therapeutic cloning allows for genetically identical organs and tissues to be harvested from the clone for the use of the organism that is cloned. "Spare parts" cloning promises many opportunities for future medical advances. What is the ontological and ethical status of spare parts, headless clones? This paper attempts to answer this question from the perspective of Aristotle's view of the soul. Aristotle's metaphysics as applied to his view of biological essences generates an ethic that can contribute to moral reasoning regarding the use of headless spare parts clones. The task of this paper is to show the implications that Aristotle's view of the soul, if it is true, would have on the ethics of headless, spare parts cloning.

  3. Ethical issues in cloning.

    PubMed

    Satris, S

    2000-01-01

    There is great public concern with the ethics of human cloning. This paper briefly examines some of what I identify as pseudo-problems or myths associated with cloning, and some of the more substantial ethical concerns.

  4. Genetically Modified Flax Expressing NAP-SsGT1 Transgene: Examination of Anti-Inflammatory Action

    PubMed Central

    Matusiewicz, Magdalena; Kosieradzka, Iwona; Zuk, Magdalena; Szopa, Jan

    2014-01-01

    The aim of the work was to define the influence of dietary supplementation with GM (genetically modified) GT#4 flaxseed cake enriched in polyphenols on inflammation development in mice liver. Mice were given ad libitum isoprotein diets: (1) standard diet; (2) high-fat diet rich in lard, high-fat diet enriched with 30% of (3) isogenic flax Linola seed cake; and (4) GM GT#4 flaxseed cake; for 96 days. Administration of transgenic and isogenic seed cake lowered body weight gain, of transgenic to the standard diet level. Serum total antioxidant status was statistically significantly improved in GT#4 flaxseed cake group and did not differ from Linola. Serum thiobarbituric acid reactive substances, lipid profile and the liver concentration of pro-inflammatory cytokine tumor necrosis factor-α were ameliorated by GM and isogenic flaxseed cake consumption. The level of pro-inflammatory cytokine interferon-γ did not differ between mice obtaining GM GT#4 and non-GM flaxseed cakes. The C-reactive protein concentration was reduced in animals fed GT#4 flaxseed cake and did not differ from those fed non-GM flaxseed cake-based diet. Similarly, the liver structure of mice consuming diets enriched in flaxseed cake was improved. Dietetic enrichment with GM GT#4 and non-GM flaxseed cakes may be a promising solution for health problems resulting from improper diet. PMID:25247574

  5. Genetically modified flax expressing NAP-SsGT1 transgene: examination of anti-inflammatory action.

    PubMed

    Matusiewicz, Magdalena; Kosieradzka, Iwona; Zuk, Magdalena; Szopa, Jan

    2014-09-22

    The aim of the work was to define the influence of dietary supplementation with GM (genetically modified) GT#4 flaxseed cake enriched in polyphenols on inflammation development in mice liver. Mice were given ad libitum isoprotein diets: (1) standard diet; (2) high-fat diet rich in lard, high-fat diet enriched with 30% of (3) isogenic flax Linola seed cake; and (4) GM GT#4 flaxseed cake; for 96 days. Administration of transgenic and isogenic seed cake lowered body weight gain, of transgenic to the standard diet level. Serum total antioxidant status was statistically significantly improved in GT#4 flaxseed cake group and did not differ from Linola. Serum thiobarbituric acid reactive substances, lipid profile and the liver concentration of pro-inflammatory cytokine tumor necrosis factor-α were ameliorated by GM and isogenic flaxseed cake consumption. The level of pro-inflammatory cytokine interferon-γ did not differ between mice obtaining GM GT#4 and non-GM flaxseed cakes. The C-reactive protein concentration was reduced in animals fed GT#4 flaxseed cake and did not differ from those fed non-GM flaxseed cake-based diet. Similarly, the liver structure of mice consuming diets enriched in flaxseed cake was improved. Dietetic enrichment with GM GT#4 and non-GM flaxseed cakes may be a promising solution for health problems resulting from improper diet.

  6. Sleep quality in efavirenz-treated Chinese HIV patients - comparing between GT and GG genotype of CYP2B6-516 G/T polymorphisms.

    PubMed

    Lee, Shui Shan; To, Kin Wang; Lee, Man Po; Wong, Ngai Sze; Chan, Denise P C; Li, Patrick C K; Cheung, Siu Wai; Chan, Raphael C Y

    2014-03-01

    Seventy-two adult Chinese HIV-positive treatment-naïve patients were recruited in a study to evaluate prospectively the associations between CYP2B6 516 G/T polymorphisms and sleep quality following treatment with an efavirenz-based regimen. Overall, the patients gave an allelic frequency of 0.3 for CYP2B6 516 T, and a genotype frequency of 9.4% for TT. Compared to GG, GT gave a higher median value of plasma efavirenz level at four weeks (3.77 mg/L vs 2.59 mg/L, p < 0.001) and 12 months (3.57 mg/L vs 2.97 mg/L, p = 0.026). Using generalised estimating equations analysis to track the variance over time, there was poorer Pittsburgh Sleep Quality Index in GT compared to GG, while GT was associated with a higher efavirenz level of >4 mg/L. There was however no difference in the component sleep scores nor was there direct association between sleep quality and plasma efavirenz levels. The results suggested that CYP2B6 genotype was associated with different patterns of sleep problems, further investigation of which is warranted with the objective of optimizing therapy with efavirenz-based regimens.

  7. Cloning in reproductive medicine.

    PubMed

    Illmensee, K

    2001-08-01

    This review article summarizes the historical development of mammalian cloning, presents current advances and presumed risk factors in the field of reproductive cloning, discusses possible clinical applications of therapeutic and diagnostic cloning and outlines prospective commercial trends in pharmaceutical cloning. Predictable progress in biotechnology and stem cell engineering should prove to be advantageous for patients' health and for novel benefits in reproductive and regenerative medicine.

  8. Acetic-acid-mediated miscibility toward electrospinning homogeneous composite nanofibers of GT/PCL.

    PubMed

    Feng, Bei; Tu, Hongbin; Yuan, Huihua; Peng, Hongju; Zhang, Yanzhong

    2012-12-10

    In tissue engineering research, there has recently been considerable interest in using electrospun biomimetic nanofibers of hybrids, in particular, from natural and synthetic polymers for engineering different tissues. However, phase separation between a pair of much dissimilar polymers might give rise to detrimental influences on both the electrospinning process and the resultant fiber performance. A representative natural-synthetic hybrid of gelatin (GT) and polycaprolactone (PCL) (50:50) was employed to study the phase separation behavior in electrospinning of the GT/PCL composite fibers. Using trifluoroethanol (TFE) as the cosolvent of the two polymers, observation of visible sedimentation and flocculation from dynamic light scattering analysis of the GT/PCL/TFE mixture both showed that phase separation does occur in just a few hours. This consequently led to gradually deteriorated fiber morphologies (e.g., splash, fiber bonding, and varied fiber size) over time during electrospinning GT/PCL. Quantitative analysis also indicated that the ratio of GT to PCL in the resultant GT/PCL fibers was altered over time. To address the phase separation related issues, a tiny amount (<0.3%) of acetic acid was introduced to improve the miscibility, which enabled the originally turbid solution to become clear immediately and to be single-phase stable for more than 1 week. Nanofibers thus obtained also appeared to be thinner, smooth, and homogeneous with enhanced performance in wettability and mechanical properties. Given the versatility and widely uses of the electrospun GT/PCL and other similar natural-synthetic hybrid systems in constructing tissue-engineered scaffolds, this work may offer a facile and effective approach to achieve finer and compositionally homogeneous hybrid nanofibers for effective applications.

  9. Quick and clean cloning.

    PubMed

    Thieme, Frank; Marillonnet, Sylvestre

    2014-01-01

    Identification of unknown sequences that flank known sequences of interest requires PCR amplification of DNA fragments that contain the junction between the known and unknown flanking sequences. Since amplified products often contain a mixture of specific and nonspecific products, the quick and clean (QC) cloning procedure was developed to clone specific products only. QC cloning is a ligation-independent cloning procedure that relies on the exonuclease activity of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. A specific feature of QC cloning is the use of vectors that contain a sequence called catching sequence that allows cloning specific products only. QC cloning is performed by a one-pot incubation of insert and vector in the presence of T4 DNA polymerase at room temperature for 10 min followed by direct transformation of the incubation mix in chemo-competent Escherichia coli cells.

  10. The activity of interleukin-4 receptor alpha-chain promoter is regulated by a GT box element.

    PubMed

    Dorado, Beatriz; Martín-Saavedra, Francisco M; Jerez, María J; Ballester, Sara

    2006-04-01

    Interleukin-4 receptor (IL-4R) is the cell surface complex through which interleukin-4 (IL-4) signals exert its critical biological effects. The alpha-chain of IL-4R is responsible for the high affinity binding of IL-4. In this report, is characterized, the 5' untranslated flanking region of murine IL-4Ralpha gene in the Th2 clone D10.G4.1. We have analyzed a DNA fragment spanning from -995 to +84 relative to the transcription start point. Mutagenesis analysis shows that, neither the previously described Stat6 (-395) nor the NFAT (-266) and NFkappaB (+25) sequences localized here, are involved in the IL-4Ralpha promoter activity. Reporter assays demonstrate that maximum transcriptional activity is achieved by the -89 to +84 sequence and this activity is independent of a TATA-like box located at -25. We have identified a GT box located at -45 as the critical element for the IL-4Ralpha promoter activity. Experiments in SL2 cells, which lack endogenous Sp proteins, show that IL-4Ralpha minimal promoter is transactivated by proteins of Sp family.

  11. Molecular cloning of a Brassica napus thiohydroximate S-glucosyltransferase gene and its expression in Escherichia coli.

    PubMed

    Marillia, Elizabeth-France; MacPherson, Jim M.; Tsang, Edward W. T.; Van Audenhove, Katrien; Keller, Wilf A.; GrootWassink, Jan W. D.

    2001-10-01

    A genomic clone encoding a thiohydroximate S-glucosyltransferase (S-GT) was isolated from Brassica napus by library screening with probes generated by PCR using degenerated primers. Its corresponding cDNA was amplified by rapid amplification of cDNA ends (RACE) PCR and also cloned by cDNA library screening. The genomic clone was 5 896 bp long and contained a 173-bp intron. At least two copies of the S-GT gene were present in B. napus. The full-length cDNA clone was 1.5 kb long and contained an open reading frame encoding a 51-kDa polypeptide. The deduced amino acid sequence shared a significant degree of homology with other glucosyltransferases characterized in other species, including a highly conserved motif within this family of enzymes corresponding to the glucose-binding domain. The recombinant protein was expressed in Escherichia coli, and the enzyme activity was tested by a biochemical assay based on the measure of glucose incorporation. The high thiohydroximate S-GT activity detected from the recombinant protein confirmed that this clone was indeed a S-glucosyltransferase.

  12. Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)

    PubMed Central

    Ovchinnikova, Olga G.; Mallette, Evan; Koizumi, Akihiko; Lowary, Todd L.; Kimber, Matthew S.

    2016-01-01

    Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. It is often involved in attaching surface polysaccharides to their lipid anchors. α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links “group 2” capsular polysaccharides to (lyso)phosphatidylglycerol. β-Kdo is also found in a small number of other bacterial polysaccharides. The structure and function of the prototypical cytidine monophosphate-Kdo–dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena. The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. PMID:27199480

  13. A thermodynamic study of waste heat recovery from GT-MHR using organic Rankine cycles

    NASA Astrophysics Data System (ADS)

    Yari, Mortaza; Mahmoudi, S. M. S.

    2011-02-01

    This paper presents an investigation on the utilization of waste heat from a gas turbine-modular helium reactor (GT-MHR) using different arrangements of organic Rankine cycles (ORCs) for power production. The considered organic Rankine cycles were: simple organic Rankine cycle (SORC), ORC with internal heat exchanger (HORC) and regenerative organic Rankine cycle (RORC). The performances of the combined cycles were studied from the point of view of first and second-laws of thermodynamics. Individual models were developed for each component and the effects of some important parameters such as compressor pressure ratio, turbine inlet temperature, and evaporator and environment temperatures on the efficiencies and on the exergy destruction rate were studied. Finally the combined cycles were optimized thermodynamically using the EES (Engineering Equation Solver) software. Based on the identical operating conditions for the GT-MHR cycle, a comparison between the three combined cycles and a simple GT-MHR cycle is also were made. This comparison was also carried out from the point of view of economics. The GT-MHR/SORC combined cycle proved to be the best among all the cycles from the point of view of both thermodynamics and economics. The efficiency of this cycle was about 10% higher than that of GT-MHR alone.

  14. Ganglioside GT1b protects human spermatozoa from hydrogen peroxide-induced DNA and membrane damage.

    PubMed

    Gavella, Mirjana; Garaj-Vrhovac, Verica; Lipovac, Vaskresenija; Antica, Mariastefania; Gajski, Goran; Car, Nikica

    2010-06-01

    We have reported previously that various gangliosides, the sialic acid containing glycosphingolipids, provide protection against sperm injury caused by reactive oxygen species (ROS). In this study, we investigated the effect of treatment of human spermatozoa with ganglioside GT1b on hydrogen peroxide (H(2)O(2))-induced DNA fragmentation and plasma membrane damage. Single-cell gel electrophoresis (Comet assay) used in the assessment of sperm DNA integrity showed that in vitro supplemented GT1b (100 microm) significantly reduced DNA damage induced by H(2)O(2) (200 microm) (p < 0.05). Measurements of Annexin V binding in combination with the propidium iodide vital dye labelling demonstrated that the spermatozoa pre-treated with GT1b exhibited a significant increase (p < 0.05) in the percentage of live cells with intact membrane and decreased phosphatidylserine translocation after exposure to H(2)O(2). Flow cytometry using the intracellular ROS-sensitive fluorescence dichlorodihydrofluorescein diacetate dye employed to investigate the transport of the extracellularly supplied H(2)O(2) into the cell interior revealed that ganglioside GT1b completely inhibited the passage of H(2)O(2) through the sperm membrane. These results suggest that ganglioside GT1b may protect human spermatozoa from H(2)O(2)-induced damage by rendering sperm membrane more hydrophobic, thus inhibiting the diffusion of H(2)O(2) across the membrane.

  15. [Construction of genomic library of L. interrogans serovar lai using lambda gt11 as the vector and a study of recombiant plasmid pDL121].

    PubMed

    Liu, H; Dai, B; Jing, B; Wu, W; Li, S; Fang, Z; Zhao, H; Ye, D; Yan, R; Liu, J; Song, S; Yang, Y; Zhang, Y; Liu, F; Tu, Y; Yang, H; Huang, Z; Liang, L; Hu, L; Zhao, M

    1997-03-01

    A genomic library of L. interrogans serovar lai strain 017 has been constructed using lambda gt11 as the vector. DNA was partially digested by two blunt-end restriction enzymes, then methylated with EcoR I methylase; after EcoR I linker was added to the DNA, the linker-ended DNA was ligated to the dephosphorylated EcoR I digested lambda gt11 arms. The recombined DNA was packaged in vitro, and used to transduct E. coli Y1090 for amplification. There were 2.1 x 10(6) recombinant bacteriophages as recognized by their ability to form white plaques plated on Lac host in the presence of both IPTG and X-Ga1. A positive clone, designated lambda DL12, was screened with a rabbit anti-serum against L. interrogans serovar lai from the genomic library. The DNA from lambda DL12 was subcloned into plasmid pUC18. A recombinant (designated as pDL121) was obtained. SDS-PAGE analysis indicated that a 23 kd was expressed in E. coli JM 103 harboring pDL121. Western blotting analysis showed that a specific protein band molecular weight of 23 kd could be recognized by the rabbit antiserum against L. interrogans serovar lai strain 017.

  16. [MxA gene-88 G/T polymorphism influences the outcomes of HBV infection].

    PubMed

    Yin, Si-chun; Peng, Xiao-mou; Gu, Lin; Huang, Yang-su; Gao, Zhi-liang

    2006-06-01

    To study the relationship between a G/T substitution at position -88 of myxovirus resistance-1 gene (MxA) and the self-limiting or chronic infection of HBV. Blood samples from 100 patients with self-limiting HBV infection (positive anti-HBs and anti-HBc) and from 340 patients with chronic HBV infection were collected. MxA-88 G/T polymorphism was typed using a protocol based on competitively differentiated-polymerase chain reaction. For statistical analysis, odds ratio and chi-square test were used. The detective rate of G/G genotype (low expression genotype) of MxA-88 G/T was 50.2% (221/440), those of T/T genotype (high expression genotype) and G/T heterozygous genotype were 5.5% (24/440) and 44.3% (195/440). Compared to patients with chronic infection, patients with self-limiting infection had lower frequency of G/G genotype (41.0% vs 52.9%, P < 0.05) or G allele (62.5% vs 75.9%, P < 0.01) and had higher frequency of T/T genotype (16.0% vs 2.4%, P < 0.01) or T allele (37.5% vs 24.1%, P < 0.01), but there was no significant difference in the G/T heterozygous genotype. MxA gene -88 G/T polymorphism influences the natural outcomes of HBV infection to some extent. This SNP of MxA gene may be used as a clinical prognostic marker of HBV infection.

  17. Nuclear transfer and cloning.

    PubMed

    Wolf, D P

    2001-10-01

    The use of nuclear transfer in human reproductive and therapeutic cloning is reviewed with attention on the origins of this technology from its evolution to the present. The successes and limitations of mammalian reproductive cloning are itemized. A case is made against the use of human reproductive cloning to reproduce an existing person, based on the unacceptable risks to the embryo, fetus, or newborn. However, support is extended for human therapeutic cloning involving the derivation and use of embryonic stem cells to treat human disease.

  18. [Cloning--ethical aspects].

    PubMed

    Munzarová, M

    2004-01-01

    Ethical problems related to cloning are discussed on three model situations: cloning of human beings (for example by utilizing the techniques of embryo splitting or nuclear transfer), use of embryonic cells in cloning techniques and cloning of nonembryonic cells. The first situation is strictly condemned, the second has been examined up present (it should be condemned as well) and the third is--under certain conditions--fully acceptable. The issue is discussed from the point of view of relevant Council of Europe documents as well.

  19. Optimizing the G/T ratio of the DSS-13 34-meter beam-waveguide antenna

    NASA Technical Reports Server (NTRS)

    Esquivel, M. S.

    1992-01-01

    Calculations using Physical Optics computer software were done to optimize the gain-to-noise temperature (G/T) ratio of DSS-13, the DSN's 34-m beam-waveguide antenna, at X-band for operation with the ultra-low-noise amplifier maser system. A better G/T value was obtained by using a 24.2-dB far-field-gain smooth-wall dual-mode horn than by using the standard X-band 22.5-dB-gain corrugated horn.

  20. Optimizing the G/T ratio of the DSS-13 34-meter beam-waveguide antenna

    NASA Technical Reports Server (NTRS)

    Esquivel, M. S.

    1992-01-01

    Calculations using Physical Optics computer software were done to optimize the gain-to-noise-temperature (G/T) ratio of Deep Space Station (DSS)-13, the Deep Space Network's (DSN's) 34-m beam-waveguide antenna, at X-band for operation with the ultra-low-noise amplifier maser system. A better G/T value was obtained by using a 24.2-dB far-field-gain smooth-wall dual-mode horn than by using the standard X-band 22.5-dB-gain corrugated horn.

  1. LU60645GT and MA132843GT Catalogues of Lunar and Martian Impact Craters Developed Using a Crater Shape-based Interpolation Crater Detection Algorithm for Topography Data

    NASA Technical Reports Server (NTRS)

    Salamuniccar, Goran; Loncaric, Sven; Mazarico, Erwan Matias

    2012-01-01

    For Mars, 57,633 craters from the manually assembled catalogues and 72,668 additional craters identified using several crater detection algorithms (CDAs) have been merged into the MA130301GT catalogue. By contrast, for the Moon the most complete previous catalogue contains only 14,923 craters. Two recent missions provided higher-quality digital elevation maps (DEMs): SELENE (in 1/16° resolution) and Lunar Reconnaissance Orbiter (we used up to 1/512°). This was the main motivation for work on the new Crater Shape-based interpolation module, which improves previous CDA as follows: (1) it decreases the number of false-detections for the required number of true detections; (2) it improves detection capabilities for very small craters; and (3) it provides more accurate automated measurements of craters' properties. The results are: (1) LU60645GT, which is currently the most complete (up to D>=8 km) catalogue of Lunar craters; and (2) MA132843GT catalogue of Martian craters complete up to D>=2 km, which is the extension of the previous MA130301GT catalogue. As previously achieved for Mars, LU60645GT provides all properties that were provided by the previous Lunar catalogues, plus: (1) correlation between morphological descriptors from used catalogues; (2) correlation between manually assigned attributes and automated measurements; (3) average errors and their standard deviations for manually and automatically assigned attributes such as position coordinates, diameter, depth/diameter ratio, etc; and (4) a review of positional accuracy of used datasets. Additionally, surface dating could potentially be improved with the exhaustiveness of this new catalogue. The accompanying results are: (1) the possibility of comparing a large number of Lunar and Martian craters, of e.g. depth/diameter ratio and 2D profiles; (2) utilisation of a method for re-projection of datasets and catalogues, which is very useful for craters that are very close to poles; and (3) the extension of the

  2. Activation of clones producing self-reactive antibodies by foreign antigen and antiidiotype antibody carrying the internal image of the antigen.

    PubMed Central

    Bailey, N C; Fidanza, V; Mayer, R; Mazza, G; Fougereau, M; Bona, C

    1989-01-01

    Because we found in previous work that a high fraction of antibodies exhibiting various specificities bound to glutamic acid 50-tyrosine50 homopolymer (GT) and expressed pGAT cross-reactive idiotype (IdX), we studied the activation of clones producing multireactive antibodies in 1-mo-old MRL/lpr and C3H/HeJ mice bearing VHJ haplotype. The activation of such clones was studied after mice were immunized with GT in CFA, HP20 (an anti-Id MAb carrying the internal image of GT in the D region), and a synthetic peptide corresponding to the D segment of HP20. Our results indicate that immunized mice produced both GT- and self-reactive antibodies. Study of the immunochemical properties of MAb showed that they exhibit multispecific properties and bind with similar-affinity constants to GT or self-antigens such as DNA, Smith antigen (Sm), and IgG2a. An important fraction of antibodies obtained from MRL/lpr mice immunized with HP20 expressed pGAT IdX and some of these antibodies share IdX expressed on anti-DNA, Sm, and rheumatoid factor (RFs) antibodies. The hybridomas producing multispecific autoantibodies use heavy-chain- (VH) and light-chain-variable region (VK) genes from various V gene families, suggesting that they do not derive from the pool of GAT precursors. Sequencing of VH and VK genes of two antibodies show that they can use closely related VHJ558, unmutated VK1, or different VK genes than those used by anti-GT antibodies. Our data demonstrate that clones producing antibodies binding to GT and self-antigens with similar-affinity constants can be activated by foreign or anti-Id antibodies carrying the internal image of the antigen or even by a synthetic peptide corresponding to the D segment of anti-Id antibodies. Images PMID:2760212

  3. Molecular cloning and amino acid sequence of leukotriene A4 hydrolase.

    PubMed Central

    Funk, C D; Rådmark, O; Fu, J Y; Matsumoto, T; Jörnvall, H; Shimizu, T; Samuelsson, B

    1987-01-01

    A cDNA clone corresponding to leukotriene A4 hydrolase was isolated from a human lung lambda gt11 expression library by immunoscreening with a polyclonal antiserum. Several additional clones from human lung and placenta cDNA lambda g11 libraries were obtained by plaque hybridization with the 32P-labeled lung cDNA clone. One of these clones has an insert of 1910 base pairs that contains the complete protein-coding region. From the deduced primary structure, leukotriene A4 hydrolase is a 610 amino and protein with a calculated molecular weight of 69,140. No apparent homologies with microsomal epoxide hydrolases were found. RNA blot analysis indicated substantial amounts of a discrete mRNA of approximately equal to 2250 nucleotides in lung tissue and leukocytes. Images PMID:2821541

  4. CDNA CLONING OF FATHEAD MINNOW (PIMEPHALES PROMELAS) ESTROGEN AND ANDROGEN RECEPTORS FOR USE IN STEROID RECEPTOR EXTRAPOLATION STUDIES FOR ENDOCRINE DISRUPTING CHEMICALS

    EPA Science Inventory

    cDNA Cloning of Fathead minnow (Pimephales promelas) Estrogen and Androgen Receptors for Use in Steroid Receptor Extrapolation Studies for Endocrine Disrupting Chemicals.

    Wilson, V.S.1,, Korte, J.2, Hartig P. 1, Ankley, G.T.2, Gray, L.E., Jr 1, , and Welch, J.E.1. 1U.S...

  5. CDNA CLONING OF FATHEAD MINNOW (PIMEPHALES PROMELAS) ESTROGEN AND ANDROGEN RECEPTORS FOR USE IN STEROID RECEPTOR EXTRAPOLATION STUDIES FOR ENDOCRINE DISRUPTING CHEMICALS

    EPA Science Inventory

    cDNA Cloning of Fathead minnow (Pimephales promelas) Estrogen and Androgen Receptors for Use in Steroid Receptor Extrapolation Studies for Endocrine Disrupting Chemicals.

    Wilson, V.S.1,, Korte, J.2, Hartig P. 1, Ankley, G.T.2, Gray, L.E., Jr 1, , and Welch, J.E.1. 1U.S...

  6. Cloning, killing, and identity.

    PubMed Central

    McMahan, J

    1999-01-01

    One potentially valuable use of cloning is to provide a source of tissues or organs for transplantation. The most important objection to this use of cloning is that a human clone would be the sort of entity that it would be seriously wrong to kill. I argue that entities of the sort that you and I essentially are do not begin to exist until around the seventh month of fetal gestation. Therefore to kill a clone prior to that would not be to kill someone like you or me but would be only to prevent one of us from existing. And even after one of us begins to exist, the objections to killing it remain comparatively weak until its psychological capacities reach a certain level of maturation. These claims support the permissibility of killing a clone during the early stages of its development in order to use its organs for transplantation. PMID:10226909

  7. Comparison of Yamax pedometer and GT3X accelerometer steps in a free-living sample

    USDA-ARS?s Scientific Manuscript database

    Our objective was to compare steps detected by the Yamax pedometer (PEDO) versus the GT3X accelerometer (ACCEL) in free-living adults. Daily PEDO and ACCEL steps were collected from a sample of 23 overweight and obese participants (18 females; mean +/- sd: age = 52.6 +/- 8.4 yr.; body mass index = 3...

  8. The "Invisible" Gifted and Talented Bilingual Students: A Current Report on Enrollment in GT Programs

    ERIC Educational Resources Information Center

    Esquierdo, J. Joy; Arreguin-Anderson, Maria

    2012-01-01

    The issue of underrepresentation in gifted and talented (GT) programs has developed into a critical educational concern. At the core are ambiguous identification assessment practices, especially for bilingual students. To illustrate, this article reports data from the last decade that supports the underrepresentation of gifted Hispanic bilingual…

  9. The National Research Center on the Gifted and Talented (NRC/GT) Newsletter, June 1991-Winter 1997.

    ERIC Educational Resources Information Center

    Gubbins, E. Jean, Ed.; Siegle, Del L., Ed.

    1997-01-01

    These 15 newsletters from the National Research Center on the Gifted and Talented (NRC/GT) contain the following articles: (1) "National Research Needs Assessment Process" (Brian D. Reid); (2) "NRC/GT: Update of Year 2 Activities" (E. Jean Gubbins); (3) "Parents: Their Impact on Gifted Adolescents" (Julie L. Sherman);…

  10. 78 FR 29810 - Receipt of Petition for Decision That Nonconforming 2003 BMW K 1200 GT Motorcycles Are Eligible...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-21

    ... 2003 BMW K 1200 GT Motorcycles Are Eligible for Importation AGENCY: National Highway Traffic Safety... Traffic Safety Administration (NHTSA) of a petition for a decision that 2003 BMW K 1200 GT Motorcycles... (Registered Importer R-09-005) has petitioned NHTSA to decide whether non-U.S. certified 2003 BMW K 1200...

  11. On cloning human beings.

    PubMed

    de Melo-Martin, Inmaculada

    2002-06-01

    The purpose of this paper is to show that arguments for and against cloning fail to make their case because of one or both of the following reasons: 1) they take for granted customary beliefs and assumptions that are far from being unquestionable; 2) they tend to ignore the context in which human cloning is developed. I will analyze some of the assumptions underlying the main arguments that have been offered for and against cloning. Once these assumptions are critically analyzed, arguments both rejecting and supporting human cloning seem to lose weight. I will first briefly present the main arguments that have been proposed against cloning and I will argue that they fail to establish their case. In the next section I will evaluate some of the positive arguments that have been offered supporting such technology. This analysis will show that the case for cloning also fails. Finally, I will maintain that because critics and especially supporters of this technology neglect the context in which human cloning is developed and might be implemented, their arguments are far from compelling.

  12. [Human cloning or cannibalism].

    PubMed

    Sokolowski, L M

    2001-01-01

    In this article I develop the idea presented in my previous work that human cloning would be of little practical use since almost any aim that one would like to attain by multiple cloning of a concrete man or a group of people, are unattainable or it might be achieved by easier, cheaper and more efficient traditional methods. For this reason cloning of a man is unlikely to occur on a larger scale and only few people will decide to clone themselves. In this sense no social effects of human cloning will be disastrous for the human population. Yet investigations in human genetics are very important since they may provide medical applications far more important than human cloning. It is argued that the main trend of modern medicine: organ transplantation from an alien donor, will become socially dangerous in near future since the number of donors will be drastically smaller than the number of potential patients waiting for transplantations. This in turn may cause social conflicts and a form of medical cannibalism may arise. These problems and conflicts will be avoided if organ transplantation from an alien donor is replaced by organ cloning, i.e. by transplanting an organ developed from the patient.

  13. Gonadotropic hormone (GtH) receptors in the testis of the troutSalmo gairdneri: in vitro studies.

    PubMed

    Le Gac, F; Breton, B; Bougoussa, M

    1988-10-01

    A particulate fraction obtained from trout testis at the time of spermiation shows saturable binding sites for(125)I-labeled salmon gonadotropin ((125)I-GtH). Non-gonadal tissues (liver, muscle and spleen) did not demonstrate specific(125)I-GtH binding. The tracer's specific activity was determined by the self-displacement method (18 to 30 μCi/μg). Maximal specific binding ability of(125)I-GtH varied from 20 to 30% of the labelled ligand added, depending on the hormone preparation. Specific binding of(125)I-GtH to 20 mg of the testis membrane varied from 40 to 85% of the total binding depending on the method of membrane prepratation, and was competitively inhibited by concentrations of unlabelled GtH ranging from ca 1 to 1000 ng/ml of incubate. Gonadotropin of mammalian origin, ovine TSH or salmon prolactin competed only weakly, or not at all, for testicular gonadotropin binding sites (relative potencies s-GtH>FSH=hCG>s-PRL>bTSH). Scatchard analysis of equilibrium binding studies shows that saturable gonadotropin binding was due to a class of high affinity binding sites (sites I Ka≊3×10(10) M(-1)) and possibly to a second class of lower affinity binding sites (sites II Ka=5 to 14×10(8) M(-1)). The binding capacity of sites I, as measured in enriched membrane preparations, was 45±18 fmoles/g of testis during the period of spermiation. The concentration of GtH required to obtain half maximal displacement of(125)I-GtH in the binding studies was of the same order of magnitude as the apparent ED50 for GtH stimulation of 11-Cetotestosterone (11KT) secretion by trout testesin vitro. Mammalian LH and FSH were 100 to 1000 folds less potent than salmor GtH to increase 11 KT secretion.

  14. Do Managers Clone Themselves?

    ERIC Educational Resources Information Center

    Baron, Alma S.

    1981-01-01

    A recent questionnaire survey provides statistics on male managers' views of female managers. The author recommends that male managers break out of their cloning behavior and that the goal ought to be a plurality in management. (Author/WD)

  15. Clone clustering by hybridization

    SciTech Connect

    Milosavijevic, A.; Zeremski, M.; Paunesku, T.

    1995-05-01

    DNA sequencing by hybridization (SBH) Format 1 technique is based on experiments in which thousands of short oligomers are consecutively hybridized with dense arrays of clones. In this paper the authors present the description of a method for obtaining hybridization signatures for individual clones that guarantees reproducibility despite a wide range of variations in experimental circumstances, a sensitive method for signature comparison at prespecified significance levels, and a clustering algorithm that correctly identifies clusters of significantly similar signatures. The methods and the algorithm have been verified experimentally on a control set of 422 signatures that originate from 9 distinct clones of known sequence. Experiments indicate that only 30 to 50 oligomer probes suffice for correct clustering. This information about the identity of clones can be used to guide both genomic and cDNA sequencing by SBH or by standard gel-based methods. 12 refs., 5 figs., 2 tabs.

  16. Do Managers Clone Themselves?

    ERIC Educational Resources Information Center

    Baron, Alma S.

    1981-01-01

    A recent questionnaire survey provides statistics on male managers' views of female managers. The author recommends that male managers break out of their cloning behavior and that the goal ought to be a plurality in management. (Author/WD)

  17. Statement on Human Cloning

    MedlinePlus

    ... form Search American Association for the Advancement of Science Statement on Human Cloning Tweet The American Association for the Advancement of Science (AAAS) recognizes the intense debates within our society ...

  18. Statement on Human Cloning

    MedlinePlus

    ... form Search American Association for the Advancement of Science Statement on Human Cloning Tweet The American Association for the Advancement of Science (AAAS) recognizes the intense debates within our society ...

  19. Twins: A cloning experience.

    PubMed

    Prainsack, Barbara; Spector, Tim D

    2006-11-01

    Drawing upon qualitative interviews with monozygotic (identical) twins sharing 100% of their genes, and with dizygotic (fraternal) twins and singletons as control groups, this paper explores what it means to be genetically identical. (The twins interviewed were from the TwinsUK register in London.) In the context of the ongoing debate on human reproductive cloning, it examines questions such as: To what extent do identical twins perceive their emotional and physical bond to be a result of their genetic makeup? What would they think if they had been deliberately created genetically identical? How would they feel about being genetically identical to a person who was born a few years earlier or later? First, our respondents ascribed no great significance to the role of genes in their understanding of what it means to be identical twins. Second, the opinion that human reproductive cloning would "interfere with nature", or "contradict God's will", was expressed by our respondents exclusively on the abstract level. The more our respondents were able to relate a particular invented cloning scenario to their own life-worlds, the lower the prevalence of the argument. Third, for all three groups of respondents, the scenario of having been born in one of the other groups was perceived as strange. Fourth, the aspect that our respondents disliked about cloning scenarios was the potential motives of the cloners. Without equating monozygotic twins directly with "clones", these results from "naturally" genetically identical individuals add a new dimension to what a future cloning situation could entail: The cloned person might possibly (a) perceive a close physical and emotional connection to the progenitor as a blessing; (b) suffer from preconceptions of people who regard physical likeness as a sign of incomplete individuality; and (c) perceive the idea of not having been born a clone of a particular person as unpleasant.

  20. GT-6

    NASA Image and Video Library

    1965-12-16

    S65-61825 (16 Dec. 1965) --- Astronauts Walter M. Schirra Jr. (left), command pilot, and Thomas P. Stafford, pilot, speak to crewmen onboard the aircraft carrier USS Wasp after successful recovery of the Gemini-6 spacecraft. Note the cake with a model of the Gemini spacecraft in its center, which is positioned in front of the astronauts. Photo credit: NASA or National Aeronautics and Space Administration

  1. GT-6

    NASA Image and Video Library

    1965-12-16

    S65-61888 (16 Dec. 1965) --- Crewmen of the aircraft carrier USS Wasp gather on deck to watch the recovery of the Gemini-6 spacecraft and astronauts. The Gemini spacecraft is being hoisted along the side of the ship by crane. Photo credit: NASA or National Aeronautics and Space Administration

  2. Cloning and characterization of a glucosyltransferase from Crocus sativus stigmas involved in flavonoid glucosylation

    PubMed Central

    Moraga, Ángela Rubio; Mozos, Almudena Trapero; Ahrazem, Oussama; Gómez-Gómez, Lourdes

    2009-01-01

    Background Flavonol glucosides constitute the second group of secondary metabolites that accumulate in Crocus sativus stigmas. To date there are no reports of functionally characterized flavonoid glucosyltransferases in C. sativus, despite the importance of these compounds as antioxidant agents. Moreover, their bitter taste makes them excellent candidates for consideration as potential organoleptic agents of saffron spice, the dry stigmas of C. sativus. Results Using degenerate primers designed to match the plant secondary product glucosyltransferase (PSPG) box we cloned a full length cDNA encoding CsGT45 from C. sativus stigmas. This protein showed homology with flavonoid glucosyltransferases. In vitro reactions showed that CsGT45 catalyses the transfer of glucose from UDP_glucose to kaempferol and quercetin. Kaempferol is the unique flavonol present in C. sativus stigmas and the levels of its glucosides changed during stigma development, and these changes, are correlated with the expression levels of CsGT45 during these developmental stages. Conclusion Findings presented here suggest that CsGT45 is an active enzyme that plays a role in the formation of flavonoid glucosides in C. sativus. PMID:19695093

  3. Association between 894G>T endothelial nitric oxide synthase gene polymorphisms and metabolic syndrome.

    PubMed

    Piccoli, Jacqueline C Escobar; Gottlieb, Maria Gabriela Valle; Castro, Luciano; Bodanese, Luiz Carlos; Manenti, Euler Roberto Fernandes; Bogo, Mauricio Reis; Peres, Alessandra; Rocha, Maria Izabel U M da; Cruz, Ivana Beatrice Mânica da

    2008-11-01

    Metabolic syndrome (MS) is a cluster of cardiovascular risk factors such as hypertension, dyslipidemia, obesity and type II diabetes. Here, we performed a case-control study analyzing the association between 894G>T endothelial nitric oxide synthase gene polymorphism (NOS3) and MS in 616 subjects. Genotype frequencies were TT= 9.3%, GG= 37.2 and TG= 53.6% and the allelic frequencies were T=0.36 and G= 0.64. We observed a higher TT genotype frequency in the male MS group than control subjects (p=0.02), independent of other variables. We found an association between hypertension and TT genotype in females. Our data suggests that 894G>T plays a significant role in the mechanistic interaction between metabolic risk such as hypertension and MS, although sex-related differences may exist.

  4. Gemini Program Mission Report for Gemini-Titan 1 (GT-1)

    NASA Technical Reports Server (NTRS)

    1964-01-01

    The Gemini-Titan 1 (GT-1) space vehicle was comprised of the Gemini spacecraft and the Gemini launch vehicle. The Gemini launch vehicle is a two-stage modified Titan II ICBM. The major modifications are the addition of a malfunction detection system and a secondary flight controls system. The Gemini spacecraft, designed to carry a crew of two men on earth orbital and rendezvous missions, was unmanned for the flight reported herein (GT-1). There were no complete Gemini flight systems on board; however, the C-band transponder and telemetry transmitters were Gemini flight subsystems. Dummy equipment, having a mass and moment of inertia equal to flight system equipment, was installed in the spacecraft. The Spacecraft was instrumented to obtain data on spacecraft heating, structural loading, vibration, sound pressure levels, and temperature and pressure during the launch phase.

  5. Alloreactive T cell clones.

    PubMed

    Fitch, F W

    1984-01-01

    T cell clones are useful models for studying lymphocyte function both at the level of the individual cell and in interacting systems. Murine cytolytic and non- cytolyic T cell clones have been obtained with relative ease, and the particular procedure used to derive and maintain T cell clones may influence profoundly the characteristics of the resulting cells. The method of choice depends on the specific question to be asked. Although some clones have characteristics that would have been expected on the basis of results observed with bulk cell populations, other clones have rather unexpected properties. Although most T cell clones appear to be either cytolytic or non-cytolytic, this distinction is not always absolute. A high proportion of both cytolytic and non-cytolytic T cell clones have dual reactivity. This is true for cells which by other criteria appear to be true clones. The frequency of such cells is high enough to suggest that most if not all T cells may have reactivity for more than one antigenic determinant or that antigenic determinants recognized by T cells are shared widely and unexpectedly. It is not clear whether one or two different antigen receptors account for such dual reactivity. The nature of the T cell receptor for antigen remains obscure. T cell clones, because of their homogeneous nature, should make it easier to answer these important immunological questions. Although it remains to be determined how many distinct molecules account for the numerous biological activities found in the culture supernatants from antigen-stimulated T cell clones, it is clear that these factors influence several different types of cells that are involved directly and indirectly in immune responses. IL-2 stimulates both cytolytic and non-cytolytic T cells to proliferate. BCSF causes polyclonal activation of B cells, and there may be other factors which influence B cell responses to antigenic stimulation. IL-3 apparently stimulates maturation of immature T cells

  6. Observing Campaign for Potential Deep Impact Flyby Target 163249 (2002 GT)

    NASA Technical Reports Server (NTRS)

    Pittichova, Jana; Chesley, S. R.; Abell, P. A.; Benner, L. A. M.

    2012-01-01

    The Deep Impact spacecraft is currently on course for a Jan. 4, 2020 flyby of the sub-kilometer near-Earth asteroid 163249 (2002 GT). The re-targeting will be complete with a final small maneuver scheduled for Oct. 4, 2012. 2002 GT, which is also designated as a Potentially Hazardous Asteroid (PHA), has a well-determined orbit and is approx 800 m in diameter (H=18.3). Little more is known about the nature of this object, but in mid-2013 it will pass near the Earth, affording an exceptional opportunity for ground-based characterization. At this apparition 2002 GT will be in range of Arecibo. In addition to Doppler measurements, radar delay observations with precisions of a few microseconds are expected and have a good chance of revealing whether the system is binary or not. The asteroid will be brighter than 16th mag., which will facilitate a host of observations at a variety of wavelengths. Light curve measurements across a wide range of viewing perspectives will reveal the rotation rate and ultimately lead to strong constraints on the shape and pole orientation. Visible and infrared spectra will constrain the mineralogy, taxonomy, albedo and size. Along with the radar observations, optical astrometry will further constrain the orbit, both to facilitate terminal guidance operations and to potentially reveal nongravitational forces acting on the asteroid. Coordinating all of these observations will be a significant task and we encourage interested observers to collaborate in this effort. The 2013 apparition of 2002 GT represents a unique opportunity to characterize a potential flyby target, which will aid interpretation of the high-resolution flyby imagery and aid planning and development of the flyby imaging sequence. The knowledge gained from this flyby will be highly relevant to the human exploration program at NASA, which desires more information on the physical characteristics of sub-kilometer near-Earth asteroids.

  7. The DBA Analysis of One Control Rod Withdrawal Out of the HTR-10GT Core

    SciTech Connect

    Mingang Lang; Yujie Dong

    2006-07-01

    The 10 MW High Temperature Gas Cooled Test Reactor (HTR-10) has been built in Institute of Nuclear and New Energy Technology (INET) and has been operating successfully since the beginning of 2003. The core outlet temperature of HTR-10 is 700 deg. C. To verify the technology of gas-turbine direct cycle, INET has planned to increase its core outlet temperature to 750 deg. C and use a helium gas turbine instead of the steam generator (then the reactor is called HTR-10GT). Though HTR-10 has good intrinsic safety, the design basic accidents and beyond design basic accidents of HTR10-GT must be analyzed according to China's nuclear regulations due to changed operation parameters. THERMIX code system is used to study the accident on one control rod withdrawal out of the core by a mistake. After a control rod in the side reflector was withdrawn out at a speed of 1 cm/s by a mistake, a positive reactivity was inserted and the reactor power increased and the temperature of the core increased. When the neutron flux of power measuring range exceeded 123% and the core outlet temperature was lager than 800 deg. C, the reactor was scrammed. During the accident sequence the maximum fuel temperature was 1200.9 deg. C. It was lower than the fuel temperature limitation of 1230 deg. C. The paper compares the analysis result of HTR10-GT to those of HTR-10. The results shows that the HTR-10GT is still safe during the accident though its operating temperature is higher than HTR-10 when the fuel safety limits are the same. (authors)

  8. THE PERFORMANCE OF SMDS DIESEL FUEL MANUFACTURED BY SHELL'S GtL TECHNOLOGY

    SciTech Connect

    Clark, Richard H.

    2000-08-20

    The Royal Dutch/Shell Group's (Shell's) Gas to Liquids (GtL) technology, better known as the Shell Middle Distillate Synthesis (SMDS) process, converts natural gas into diesel and other products via a modem improved Fisher-Tropsch synthesis. The diesel cut has very good cetane quality, low density, and virtually no sulphur and aromatics; such properties make it valuable as a diesel fuel with lower emissions than conventional automotive gas oil.

  9. Observing Campaign for Potential Deep Impact Flyby Target 163249 (2002 GT)

    NASA Technical Reports Server (NTRS)

    Pittichova, Jana; Chesley, S. R.; Abell, P. A.; Benner, L. A. M.

    2012-01-01

    The Deep Impact spacecraft is currently on course for a Jan. 4, 2020 flyby of the sub-kilometer near-Earth asteroid 163249 (2002 GT). The re-targeting will be complete with a final small maneuver scheduled for Oct. 4, 2012. 2002 GT, which is also designated as a Potentially Hazardous Asteroid (PHA), has a well-determined orbit and is approx 800 m in diameter (H=18.3). Little more is known about the nature of this object, but in mid-2013 it will pass near the Earth, affording an exceptional opportunity for ground-based characterization. At this apparition 2002 GT will be in range of Arecibo. In addition to Doppler measurements, radar delay observations with precisions of a few microseconds are expected and have a good chance of revealing whether the system is binary or not. The asteroid will be brighter than 16th mag., which will facilitate a host of observations at a variety of wavelengths. Light curve measurements across a wide range of viewing perspectives will reveal the rotation rate and ultimately lead to strong constraints on the shape and pole orientation. Visible and infrared spectra will constrain the mineralogy, taxonomy, albedo and size. Along with the radar observations, optical astrometry will further constrain the orbit, both to facilitate terminal guidance operations and to potentially reveal nongravitational forces acting on the asteroid. Coordinating all of these observations will be a significant task and we encourage interested observers to collaborate in this effort. The 2013 apparition of 2002 GT represents a unique opportunity to characterize a potential flyby target, which will aid interpretation of the high-resolution flyby imagery and aid planning and development of the flyby imaging sequence. The knowledge gained from this flyby will be highly relevant to the human exploration program at NASA, which desires more information on the physical characteristics of sub-kilometer near-Earth asteroids.

  10. Feasibility study for SOFC-GT hybrid locomotive power part II. System packaging and operating route simulation

    NASA Astrophysics Data System (ADS)

    Martinez, Andrew S.; Brouwer, Jacob; Samuelsen, G. Scott

    2012-09-01

    This work assesses the feasibility of Solid Oxide Fuel Cell-Gas Turbine (SOFC-GT) hybrid power systems for use as the prime mover in freight locomotives. The available space in a diesel engine-powered locomotive is compared to that required for an SOFC-GT system, inclusive of fuel processing systems necessary for the SOFC-GT. The SOFC-GT space requirement is found to be similar to current diesel engines, without consideration of the electrical balance of plant. Preliminary design of the system layout within the locomotive is carried out for illustration. Recent advances in SOFC technology and implications of future improvements are discussed as well. A previously-developed FORTRAN model of an SOFC-GT system is then augmented to simulate the kinematics and power notching of a train and its locomotives. The operation of the SOFC-GT-powered train is investigated along a representative route in Southern California, with simulations presented for diesel reformate as well as natural gas reformate and hydrogen as fuels. Operational parameters and difficulties are explored as are comparisons of expected system performance to modern diesel engines. It is found that even in the diesel case, the SOFC-GT system provides significant savings in fuel and CO2 emissions, making it an attractive option for the rail industry.

  11. The cobas® HCV GT is a new tool that accurately identifies Hepatitis C virus genotypes for clinical practice.

    PubMed

    Fernández-Caballero, J A; Alvarez, M; Chueca, N; Pérez, A B; García, F

    2017-01-01

    We aimed to evaluate the correct assignment of HCV genotype/subtypes 1a and 1b by cobas® HCV genotyping (GT) assay (Roche Molecular Diagnostics) compared with nonstructural protein 5B (NS5B) sequencing. Clinical samples from 153 patients submitted for HCV genotyping were studied. After genotyping with the cobas® HCV GT, sequencing of a 387 bp fragment in the NS5B gene and phylogenetic analysis was employed to compare genotyping results. Major discrepancies were defined as differences in the assigned genotype by cobas® HCV GT and NS5B sequencing (including genotype 1 subtypes 1a and 1b misclassification). Overall agreement between the cobas® HCV GT and NS5B sequencing was 98%; all the 1a, 1b, 2, 3 and 4 genotypes identified by cobas® HCV GT were concordant with NS5B sequencing. Three samples tested "indetermined" by cobas® HCV GT assay and were genotyped as 1a, 3a, and 4d by NS5B sequencing. These results indicate that the cobas® HCV GT assay correctly identifies HCV genotypes, and points out the importance of additional methods based on DNA sequencing for resolving indeterminate results.

  12. MTP -493G>T polymorphism and susceptibility to nonalcoholic fatty liver disease: a meta-analysis.

    PubMed

    Zheng, Wei; Wang, Lu; Su, Xiao; Hu, Xiao-Fang

    2014-06-01

    Microsomal transfer protein (MTP), a lipid transfer protein localized in the endoplasmic reticulum of hepatocytes and enterocytes, plays an important role in the development of nonalcoholic fatty liver disease (NAFLD). Many existing studies have demonstrated that a common polymorphism (-493G>T, rs1800591 G>T) in the MTP gene may be implicated in the development and progression of NAFLD, but individually published results are inconclusive. This meta-analysis aimed to investigate whether MTP -493G>T polymorphism may be a potential risk factor for NAFLD. We searched CISCOM, CINAHL, Web of Science, PubMed, Google Scholar, EBSCO, Cochrane Library, and CBM databases from inception through October 1, 2013. Meta-analysis was performed using the STATA 12.0 software. Eleven clinical case-control studies with a total of 636 NAFLD cases and 918 healthy controls met the inclusion criteria. Our meta-analysis results revealed that MTP -493G>T polymorphism was strongly correlated with an increased risk of NAFLD. Subgroup analysis by ethnicity suggested that MTP -493G>T polymorphism might increase individuals' susceptibility to NAFLD among both Caucasian and non-Caucasian populations. No publication bias was observed in this meta-analysis. In short, the present meta-analysis indicates that MTP -493G>T polymorphisms may contribute to individuals' susceptibility to NAFLD. Thus, MTP -493G>T polymorphism may be a valuable and practical biomarker for early detection of NAFLD.

  13. The Vitamin E Analog Gamma-Tocotrienol (GT3) and Statins Synergistically Up-Regulate Endothelial Thrombomodulin (TM)

    PubMed Central

    Pathak, Rupak; Ghosh, Sanchita P.; Zhou, Daohong; Hauer-Jensen, Martin

    2016-01-01

    Statins; a class of routinely prescribed cholesterol-lowering drugs; inhibit 3-hydroxy-3-methylglutaryl-coenzymeA reductase (HMGCR) and strongly induce endothelial thrombomodulin (TM); which is known to have anti-inflammatory; anti-coagulation; anti-oxidant; and radioprotective properties. However; high-dose toxicity limits the clinical use of statins. The vitamin E family member gamma-tocotrienol (GT3) also suppresses HMGCR activity and induces TM expression without causing significant adverse side effects; even at high concentrations. To investigate the synergistic effect of statins and GT3 on TM; a low dose of atorvastatin and GT3 was used to treat human primary endothelial cells. Protein-level TM expression was measured by flow cytometry. TM functional activity was determined by activated protein C (APC) generation assay. Expression of Kruppel-like factor 2 (KLF2), one of the key transcription factors of TM, was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). TM expression increased in a dose-dependent manner after both atorvastatin and GT3 treatment. A combined treatment of a low-dose of atorvastatin and GT3 synergistically up-regulated TM expression and functional activity. Finally; atorvastatin and GT3 synergistically increased KLF2 expression. These findings suggest that combined treatment of statins with GT3 may provide significant health benefits in treating a number of pathophysiological conditions; including inflammatory and cardiovascular diseases. PMID:27869747

  14. Molecular cloning and chromosomal localization of a novel human tracheo-bronchial mucin cDNA containing tandemly repeated sequences of 48 base pairs.

    PubMed

    Porchet, N; Nguyen, V C; Dufosse, J; Audie, J P; Guyonnet-Duperat, V; Gross, M S; Denis, C; Degand, P; Bernheim, A; Aubert, J P

    1991-03-15

    A lambda gt11 cDNA library constructed from human tracheo-bronchial mucosa was screened with a polyclonal antiserum raised to chemically deglycosylated pronase glycopeptides from human bronchial mucins. Out of 20 positives clones, one partial cDNA clone was isolated and allowed to map a novel human tracheo-bronchial mucin gene. It contains 48 nucleotide tandem repeats quite perfectly identical which encodes a protein containing about 50% of hydroxy amino-acids. This clone hybridized to polydisperse messages produced by human tracheo-bronchial and human colonic mucosae. The gene (proposed name MUC 4) from which cDNA is derived maps to chromosome 3.

  15. Thermal Emission Photometry of Deep Impact Flyby Target (163249) 2002 GT

    NASA Astrophysics Data System (ADS)

    Lim, Lucy F.; Moskovitz, N. A.; Licandro, J.; Emery, J. P.; Reddy, V.; Vilas, F.; 2002 GT Observing Team

    2013-10-01

    Near-Earth asteroid (163249) 2002 GT is now the target of a Deep Impact spacecraft flyby in Jan. 2020 (see Pittichova et al., this volume, for details of the flyby and observing campaign). Thermal emission photometry of 2002 GT was obtained from NIRI on Gemini-North in the L' and M' filters, which are centered at 3.76 and 4.68 microns respectively. J- and K-band reflectance photometry was also acquired in support of the thermal observations. The full JKL'M' set was acquired on UT 2013-Jun-13 at a solar phase angle of 53 degrees. A further set of photometry in J, K, and L' only was carried out on 2013-Jun-19 at a phase angle of 65 degrees. High water vapor conditions at Mauna Kea during this period unfortunately prevented acquisition of a second set of M' measurements. In addition, N-band photometry of 2002 GT was conducted on 2013-Jun-10 from CanariCam at the 10-meter Gran Telescopio Canarias using a beta version of the moving object guiding system. Data were acquired in three filters between 8.7 and 12.5 microns, although the limitations of the guiding are complicating the analysis. (We note that N-band observing was not offered by either Gemini or IRTF during this apparition.) Data analysis is ongoing and results will be discussed. We appreciate the efforts of the Gemini and GTC staff in support of these observing programs.

  16. ASTRONAUT EDWARD H. WHITE II - GEMINI-TITAN (GT)-IV - ZERO GRAVITY - OUTER SPACE

    NASA Image and Video Library

    2015-03-20

    S65-30427 (3 June 1965) --- Astronaut Edward H. White II, pilot for the Gemini-Titan 4 (GT-4) spaceflight, floats in the zero-gravity of space during the third revolution of the GT-4 spacecraft. White wears a specially designed spacesuit. His face is shaded by a gold-plated visor to protect him from unfiltered rays of the sun. In his right hand he carries a Hand-Held Self-Maneuvering Unit (HHSMU) that gives him control over his movements in space. White also wears an emergency oxygen chest pack; and he carries a camera mounted on the HHSMU for taking pictures of the sky, Earth and the GT-4 spacecraft. He is secured to the spacecraft by a 25-feet umbilical line and a 23-feet tether line. Both lines are wrapped together in gold tape to form one cord. Astronaut James A. McDivitt, command pilot, remained inside the spacecraft during the extravehicular activity (EVA). Photo credit: NASA EDITOR'S NOTE: Astronaut Edward H. White II died in the Apollo/Saturn 204 fire at Cape Kennedy on Jan. 27, 1967.

  17. HTGR-GT closed-cycle gas turbine: a plant concept with inherent cogeneration (power plus heat production) capability

    SciTech Connect

    McDonald, C.F.

    1980-04-01

    The high-grade sensible heat rejection characteristic of the high-temperature gas-cooled reactor-gas turbine (HTGR-GT) plant is ideally suited to cogeneration. Cogeneration in this nuclear closed-cycle plant could include (1) bottoming Rankine cycle, (2) hot water or process steam production, (3) desalination, and (4) urban and industrial district heating. This paper discusses the HTGR-GT plant thermodynamic cycles, design features, and potential applications for the cogeneration operation modes. This paper concludes that the HTGR-GT plant, which can potentially approach a 50% overall efficiency in a combined cycle mode, can significantly aid national energy goals, particularly resource conservation.

  18. Effect of ganglioside GT1b on the in vitro maturation of porcine oocytes and embryonic development

    PubMed Central

    HWANG, Seon-Ung; JEON, Yubyeol; YOON, Junchul David; CAI, Lian; KIM, Eunhye; YOO, Hyunju; KIM, Kyu-Jun; PARK, Kyu Mi; JIN, Minghui; KIM, Hyunggee; HYUN, Sang-Hwan

    2015-01-01

    Ganglioside is an acidic glycosphingolipid with sialic acids residues. This study was performed to investigate the effect and mechanism of ganglioside GT1b in porcine oocytes in the process of in vitro maturation (IVM) and preimplantation development. Metaphase II (MII) rates were significantly (P < 0.05) different between the control group and the 5 nM GT1b treatment group. Intracellular glutathione (GSH) levels in oocytes matured with 5 nM and 20 nM and GT1b decreased significantly (P < 0.05). The 10 nM group showed a significant (P < 0.05) decrease in intracellular reactive oxygen species (ROS) levels compared with the control group. Subsequently, the level of intracellular Ca2+ in oocytes treated with different concentrations of GT1b was measured. Intracellular Ca2+ was significantly (P < 0.05) increased with a higher concentration of GT1b in a dose-dependent manner. Real-time PCR was performed and showed that the expression of bradykinin 2 receptor (B2R) and calcium/calmodulin-dependent protein kinase II delta (CaMKIIδ) in cumulus cells was significantly (P < 0.05) decreased in the 20 nM GT1b treatment group. Treatment with 5 nM GT1b significantly (P < 0.05) decreased the expression of CaMKIIδ. In oocytes, treatment with 5 nM GT1b significantly (P < 0.05) decreased CaMKIIγ and POU5F1 (POU domain, class 5, transcription factor 1). However, treatment with 20 nM GT1b significantly (P < 0.05) increased the expression of POU5F1. Finally, embryonic developmental data showed no significant differences in the two experiments (parthenogenesis and in vitro fertilization). In conclusion, the results of the present study indicated that GT1b plays an important role in increasing the nuclear maturation rate and decreasing the intracellular ROS levels during IVM. However, GT1b inhibited maturation of the cytoplasm by maintaining intracellular Ca2+ in the process of oocyte maturation regardless of the cell cycle stage. Therefore, GT1b is thought to act on another mechanism

  19. Molecular cloning of human terminal deoxynucleotidyltransferase.

    PubMed Central

    Peterson, R C; Cheung, L C; Mattaliano, R J; Chang, L M; Bollum, F J

    1984-01-01

    A cDNA of the human terminal deoxynucleotidyltransferase (TdT; "terminal transferase," EC 2.7.7.31) was isolated from a human lymphoblastoid cell cDNA library in lambda gt 11 by using immunological procedures. Four inserts containing 723 to 939 base pairs were recloned in pBR322 for hybridization and preliminary sequence studies. mRNA selected by hybridization to recombinant DNA was translated to a 58-kDa peptide that specifically immunoprecipitated with rabbit antibodies to calf terminal transferase and mouse monoclonal antibody to human terminal transferase. Blot hybridization of total poly(A)+ RNA from KM3 (TdT+) cells with nick-translated pBR322 recombinant DNA detected a message of about 2000 nucleotides, sufficient to code for the 580 amino acids in the protein. mRNA from terminal transferase- cells gave no signal in hybrid selection or RNA blot hybridization. The complete sequence of the 939-base-pair insert sequence was obtained from deletions cloned in pUC8. The DNA sequence contains an open reading frame coding for 238 amino acids, about 40% of the protein. Three peptides isolated by HPLC from tryptic digests of succinylated 58-kDa calf thymus terminal transferase were sequenced, providing 20, 18, and 22 residues of peptide sequence. A search of the translated sequence of the 939-base-pair insert shows three regions beginning after arginine that have greater than 90% homology with the sequence determined from the calf thymus terminal transferase peptides. These results provide unambiguous evidence that the human terminal transferase sequence has been cloned. Images PMID:6087320

  20. Biosynthesis of the major brain gangliosides GD1a and GT1b

    PubMed Central

    Sturgill, Elizabeth R; Aoki, Kazuhiro; Lopez, Pablo HH; Colacurcio, Daniel; Vajn, Katarina; Lorenzini, Ileana; Majić, Senka; Yang, Won Ho; Heffer, Marija; Tiemeyer, Michael; Marth, Jamey D; Schnaar, Ronald L

    2012-01-01

    Gangliosides—sialylated glycosphingolipids—are the major glycoconjugates of nerve cells. The same four structures—GM1, GD1a, GD1b and GT1b—comprise the great majority of gangliosides in mammalian brains. They share a common tetrasaccharide core (Galβ1–3GalNAcβ1-4Galβ1-4Glcβ1-1′Cer) with one or two sialic acids on the internal galactose and zero (GM1 and GD1b) or one (GD1a and GT1b) α2–3-linked sialic acid on the terminal galactose. Whereas the genes responsible for the sialylation of the internal galactose are known, those responsible for terminal sialylation have not been established in vivo. We report that St3gal2 and St3gal3 are responsible for nearly all the terminal sialylation of brain gangliosides in the mouse. When brain ganglioside expression was analyzed in adult St3gal1-, St3gal2-, St3gal3- and St3gal4-null mice, only St3gal2-null mice differed significantly from wild type, expressing half the normal amount of GD1a and GT1b. St3gal1/2-double-null mice were no different than St3gal2-single-null mice; however, St3gal2/3-double-null mice were >95% depleted in gangliosides GD1a and GT1b. Total ganglioside expression (lipid-bound sialic acid) in the brains of St3gal2/3-double-null mice was equivalent to that in wild-type mice, whereas total protein sialylation was reduced by half. St3gal2/3-double-null mice were small, weak and short lived. They were half the weight of wild-type mice at weaning and displayed early hindlimb dysreflexia. We conclude that the St3gal2 and St3gal3 gene products (ST3Gal-II and ST3Gal-III sialyltransferases) are largely responsible for ganglioside terminal α2-3 sialylation in the brain, synthesizing the major brain gangliosides GD1a and GT1b. PMID:22735313

  1. Biosynthesis of the major brain gangliosides GD1a and GT1b.

    PubMed

    Sturgill, Elizabeth R; Aoki, Kazuhiro; Lopez, Pablo H H; Colacurcio, Daniel; Vajn, Katarina; Lorenzini, Ileana; Majić, Senka; Yang, Won Ho; Heffer, Marija; Tiemeyer, Michael; Marth, Jamey D; Schnaar, Ronald L

    2012-10-01

    Gangliosides-sialylated glycosphingolipids-are the major glycoconjugates of nerve cells. The same four structures-GM1, GD1a, GD1b and GT1b-comprise the great majority of gangliosides in mammalian brains. They share a common tetrasaccharide core (Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1'Cer) with one or two sialic acids on the internal galactose and zero (GM1 and GD1b) or one (GD1a and GT1b) α2-3-linked sialic acid on the terminal galactose. Whereas the genes responsible for the sialylation of the internal galactose are known, those responsible for terminal sialylation have not been established in vivo. We report that St3gal2 and St3gal3 are responsible for nearly all the terminal sialylation of brain gangliosides in the mouse. When brain ganglioside expression was analyzed in adult St3gal1-, St3gal2-, St3gal3- and St3gal4-null mice, only St3gal2-null mice differed significantly from wild type, expressing half the normal amount of GD1a and GT1b. St3gal1/2-double-null mice were no different than St3gal2-single-null mice; however, St3gal2/3-double-null mice were >95% depleted in gangliosides GD1a and GT1b. Total ganglioside expression (lipid-bound sialic acid) in the brains of St3gal2/3-double-null mice was equivalent to that in wild-type mice, whereas total protein sialylation was reduced by half. St3gal2/3-double-null mice were small, weak and short lived. They were half the weight of wild-type mice at weaning and displayed early hindlimb dysreflexia. We conclude that the St3gal2 and St3gal3 gene products (ST3Gal-II and ST3Gal-III sialyltransferases) are largely responsible for ganglioside terminal α2-3 sialylation in the brain, synthesizing the major brain gangliosides GD1a and GT1b.

  2. Potential for cloning dogs.

    PubMed

    Westhusin, M E; Burghardt, R C; Ruglia, J N; Willingham, L A; Liu, L; Shin, T; Howe, L M; Kraemer, D C

    2001-01-01

    The aim of this study was to determine whether nuclear transplantation could be used to clone a dog using donor nucleus cells collected from an adult female. Fibroblasts obtained from skin biopsies were fused with enucleated bovine or canine oocytes. The resulting cloned embryos were cultured in vitro to monitor embryonic development. A proportion of the resulting embryos was transferred into surrogate bitches for development to term. When canine oocytes were used as recipient ova for canine fibroblasts, 23% of the resulting embryos cleaved at least once after culture in vitro. Five cloned embryos were transferred into three synchronized recipient bitches, but no pregnancies resulted. When bovine oocytes were used as recipinets for canine fibroblasts, 38% cleaved to the two- to four-cell stage and 43% cleaved to the eight- to 16-cell stage. Forty-seven of these embryos were transferred into four recipient females, resulting in a single conceptus that ceased development at about day 20 of gestation. The desire for cloned dogs is considerable and will undoubtedly incite the development of successful methods for cloning companion animals. However, significant investment into additional research is required, especially in the areas of in vitro maturation of oocytes and control of the oestrous cycle of bitches.

  3. Met-ase: Cloning and distinct chromosomal location of a serine protease preferentially expressed in human natural killer cells

    SciTech Connect

    Smyth, M.J.; Trapani, J.A. ); Sayers, T.J.; Wiltrout, T. ); Powers, J.C. )

    1993-12-01

    A cDNA clone encoding a human NK serine protease was obtained by screening a [lambda]-gt10 library from the Lopez NK leukemia with the rat natural killer Met-ase (RNK-Met-1) cDNA clone. In Northern blot analysis human Met-ase (Hu-Met-1) cDNA hybridized with a 0.9-kb mRNA in two human NK leukemia cell lines, unstimulated human PBMC, and untreated purified CD3[sup [minus

  4. Extremal quantum cloning machines

    NASA Astrophysics Data System (ADS)

    Chiribella, G.; D'Ariano, G. M.; Perinotti, P.; Cerf, N. J.

    2005-10-01

    We investigate the problem of cloning a set of states that is invariant under the action of an irreducible group representation. We then characterize the cloners that are extremal in the convex set of group covariant cloning machines, among which one can restrict the search for optimal cloners. For a set of states that is invariant under the discrete Weyl-Heisenberg group, we show that all extremal cloners can be unitarily realized using the so-called double-Bell states, whence providing a general proof of the popular ansatz used in the literature for finding optimal cloners in a variety of settings. Our result can also be generalized to continuous-variable optimal cloning in infinite dimensions, where the covariance group is the customary Weyl-Heisenberg group of displacements.

  5. Extremal quantum cloning machines

    SciTech Connect

    Chiribella, G.; D'Ariano, G. M.; Perinotti, P.; Cerf, N.J.

    2005-10-15

    We investigate the problem of cloning a set of states that is invariant under the action of an irreducible group representation. We then characterize the cloners that are extremal in the convex set of group covariant cloning machines, among which one can restrict the search for optimal cloners. For a set of states that is invariant under the discrete Weyl-Heisenberg group, we show that all extremal cloners can be unitarily realized using the so-called double-Bell states, whence providing a general proof of the popular ansatz used in the literature for finding optimal cloners in a variety of settings. Our result can also be generalized to continuous-variable optimal cloning in infinite dimensions, where the covariance group is the customary Weyl-Heisenberg group of displacement000.

  6. A technique for measurement of earth station antenna G/T by radio stars and Applications Technology Satellites.

    NASA Technical Reports Server (NTRS)

    Kochevar, H. J.

    1972-01-01

    A new technique has been developed to accurately measure the G/T of a small aperture antenna using geostationary satellites and the well established radio star method. A large aperture antenna having the capability of accurately measuring its G/T by using a radio star of known power density is used to obtain an accurate G/T to use as a reference. The CNR of both the large and small aperture antennas are then measured using an Applications Technology Satellite (ATS). After normalizing the two C/N ratios to the large antenna system noise temperature the G/T or the gain G of the small aperture antenna can then be determined.

  7. To clone alone: the United Nations' Human Cloning Declaration.

    PubMed

    Isasi, Rosario M; Annas, George J

    2006-01-01

    The United Nations labored for almost four years to create a treaty governing human cloning. In 2005 that effort was abandoned, and instead the United Nations' General Assembly adopted a "Declaration on Human Cloning".

  8. An in vitro comparison of cyclic fatigue resistance of ProTaper universal and GT series x files

    PubMed Central

    Montenegro-Santillán, Ramiro; Alegre-Domingo, Teresa; Faus-Matoses, Vicente

    2013-01-01

    Objective: The aim of this study was to compare the cyclic fatigue resistance of two nickel-titanium (NiTi) endodontic instruments from ProTaper and GT series X files. Study Design: Cyclic fatigue test was realized with instruments from ProTaper: F1 and F3 (Denstply Maillefer, Ballaigues, Switzerland) and GT-X: 20.06 and 30.08 (Dentsply Tulsa Dental, Tulsa, Oklahoma, United States of America). A total of 320 instruments were rotated in 4 curved artificial canals with different angles and radius of curvature. The time and cycles to failure were calculated. The data was compared using a Mann-Whitney, Kruskall-Wallis, and Kolmogorov-Smirnov tests, with a significance level of p<0.05. Results: GT-X files rotated for a significantly longer period of time before separation occurred, thus GT-X files where more resistant to the cyclic fatigue compared with ProTaper. Conclusion: GT-X files have a greater resistance to cyclic fatigue, this fact can be caused by the use of the Ni-Ti alloy “M-Wire”. Key words:Endodontics, GT-X files, ProTaper files, cyclic fatigue. PMID:23385505

  9. Apical extrusion of thermoplasticized obturating material in canals instrumented with Profile 0.06 or Profile GT.

    PubMed

    Robinson, Mark J; McDonald, N J; Mullally, Patrick J

    2004-06-01

    The purpose of this in vitro study was to compare the extrusion of thermoplacticized gutta-percha in teeth instrumented with Profile 0.06 or Profile GT, and obturated with Thermafil Plus and Thermafil GT, respectively. A total of 120, extracted, human maxillary central incisors were divided into four equal groups. Group 1 was instrumented with Profile 0.06 and obturated with Thermafil Plus. Group 2 was instrumented with Profile 0.06 and obturated using warm vertical condensation (negative control). Group 3 was instrumented with Profile GT and obturated with Thermafil GT. Group 4 was instrumented with Profile GT and obturated like Group 2 (negative control). Extrusion was graded as present or absent. Results found 9 of 30 extruded for group 1, 1 of 30 for group 2, 15 of 30 for group 3, and 2 of 30 for group 4. The results suggest that, in vitro, Thermafil GT may be more prone to extruding gutta-percha past the apical foramen than Thermafil Plus.

  10. Oocyte-secreted factors in oocyte maturation media enhance subsequent development of bovine cloned embryos.

    PubMed

    Su, Jianmin; Wang, Yongsheng; Zhang, Lei; Wang, Bo; Liu, Jun; Luo, Yan; Guo, Zekun; Quan, Fusheng; Zhang, Yong

    2014-04-01

    Successful in vitro maturation (IVM) and oocyte quality both affect the subsequent development of cloned embryos derived from somatic-cell nuclear transfer (SCNT). Developmental competence is usually lower in oocytes matured in vitro compared with those that matured in vivo, possibly due to insufficient levels of oocyte-secreted factors (OSFs) and disrupted oocyte-cumulus communication. This study investigated the effects of OSFs secreted by denuded oocytes (DOs) during IVM on the subsequent developmental competence of cloned bovine embryos. Cumulus-oocyte complexes (COCs) from antral follicles of slaughtered-cow ovaries collected from an abattoir were divided into four groups: COCs co-cultured with and without DOs in maturation media used for SCNT, as well as COCs co-cultured with and without DOs in maturation media used for in vitro fertilization (IVF). Based on the developmental competence and embryo quality of bovine embryos generated from these four groups, we found that co-culturing the COCs with DOs enhanced the in vitro development of IVF and cloned bovine embryos, and potentially generated more high-quality cloned blastocysts that possessed locus-specific histone modifications at levels similar to in vitro-fertilized embryos. These results strongly suggest that co-culturing COCs with DOs enhances subsequent developmental competence of cloned bovine embryo. © 2014 Wiley Periodicals, Inc.

  11. The Cloning of America.

    ERIC Educational Resources Information Center

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

    Proposes that the U.S. school system purports to prize human variability, but many educators are engaged in activities that seek to homogenize students. Describes these activities, including diagnosis, labeling, ability grouping, and positive reinforcement. Presents suggestions for counselors to combat sources of cloning and self-validation. (RC)

  12. Secure the Clones

    NASA Astrophysics Data System (ADS)

    Jensen, Thomas; Kirchner, Florent; Pichardie, David

    Exchanging mutable data objects with untrusted code is a delicate matter because of the risk of creating a data space that is accessible by an attacker. Consequently, secure programming guidelines for Java stress the importance of using defensive copying before accepting or handing out references to an internal mutable object. However, implementation of a copy method (like clone()) is entirely left to the programmer. It may not provide a sufficiently deep copy of an object and is subject to overriding by a malicious sub-class. Currently no language-based mechanism supports secure object cloning. This paper proposes a type-based annotation system for defining modular copy policies for class-based object-oriented programs. A copy policy specifies the maximally allowed sharing between an object and its clone. We present a static enforcement mechanism that will guarantee that all classes fulfill their copy policy, even in the presence of overriding of copy methods, and establish the semantic correctness of the overall approach in Coq. The mechanism has been implemented and experimentally evaluated on clone methods from several Java libraries.

  13. Applications of quantum cloning

    NASA Astrophysics Data System (ADS)

    Pomarico, E.; Sanguinetti, B.; Sekatski, P.; Zbinden, H.; Gisin, N.

    2011-10-01

    Quantum Cloning Machines (QCMs) allow for the copying of information, within the limits imposed by quantum mechanics. These devices are particularly interesting in the high-gain regime, i.e., when one input qubit generates a state of many output qubits. In this regime, they allow for the study of certain aspects of the quantum to classical transition. The understanding of these aspects is the root of the two recent applications that we will review in this paper: the first one is the Quantum Cloning Radiometer, a device which is able to produce an absolute measure of spectral radiance. This device exploits the fact that in the quantum regime information can be copied with only finite fidelity, whereas when a state becomes macroscopic, this fidelity gradually increases to 1. Measuring the fidelity of the cloning operation then allows to precisely determine the absolute spectral radiance of the input optical source. We will then discuss whether a Quantum Cloning Machine could be used to produce a state visible by the naked human eye, and the possibility of a Bell Experiment with humans playing the role of detectors.

  14. The Cloning of America.

    ERIC Educational Resources Information Center

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

    Proposes that the U.S. school system purports to prize human variability, but many educators are engaged in activities that seek to homogenize students. Describes these activities, including diagnosis, labeling, ability grouping, and positive reinforcement. Presents suggestions for counselors to combat sources of cloning and self-validation. (RC)

  15. Ground reaction force estimates from ActiGraph GT3X+ hip accelerations.

    PubMed

    Neugebauer, Jennifer M; Collins, Kelsey H; Hawkins, David A

    2014-01-01

    Simple methods to quantify ground reaction forces (GRFs) outside a laboratory setting are needed to understand daily loading sustained by the body. Here, we present methods to estimate peak vertical GRF (pGRFvert) and peak braking GRF (pGRFbrake) in adults using raw hip activity monitor (AM) acceleration data. The purpose of this study was to develop a statistically based model to estimate pGRFvert and pGRFbrake during walking and running from ActiGraph GT3X+ AM acceleration data. 19 males and 20 females (age 21.2 ± 1.3 years, height 1.73 ± 0.12 m, mass 67.6 ± 11.5 kg) wore an ActiGraph GT3X+ AM over their right hip. Six walking and six running trials (0.95-2.19 and 2.20-4.10 m/s, respectively) were completed. Average of the peak vertical and anterior/posterior AM acceleration (ACCvert and ACCbrake, respectively) and pGRFvert and pGRFbrake during the stance phase of gait were determined. Thirty randomly selected subjects served as the training dataset to develop generalized equations to predict pGRFvert and pGRFbrake. Using a holdout approach, the remaining 9 subjects were used to test the accuracy of the models. Generalized equations to predict pGRFvert and pGRFbrake included ACCvert and ACCbrake, respectively, mass, type of locomotion (walk or run), and type of locomotion acceleration interaction. The average absolute percent differences between actual and predicted pGRFvert and pGRFbrake were 8.3% and 17.8%, respectively, when the models were applied to the test dataset. Repeated measures generalized regression equations were developed to predict pGRFvert and pGRFbrake from ActiGraph GT3X+ AM acceleration for young adults walking and running. These equations provide a means to estimate GRFs without a force plate.

  16. Effects of filter choice in GT3X accelerometer assessments of free-living activity.

    PubMed

    Wanner, Miriam; Martin, Brian W; Meier, Flurina; Probst-Hensch, Nicole; Kriemler, Susi

    2013-01-01

    ActiGraph accelerometers are widely used devices to objectively assess physical activity. The GT3X version has two filter options to be selected before data assessment (normal and low-frequency extension filter option). It is not clear whether the resulting physical activity levels differ depending on the choice of the filter. The aims were to compare GT3X data collected using the different filter options during free-living activities and to establish correction factors if the results were not comparable. Sixty-five participants of the population-based SAPALDIA-cohort (50.8% women, age range = 40-80 yr) wore two GT3X accelerometers with different filter selections simultaneously during 8 d. Spearman correlations, Wilcoxon rank sum tests, McNemar tests, scatter plots, and Bland-Altman plots were used to compare the data. Correction factors were established using linear regression models. Although Spearman correlations were high (r ≥ 0.93), there were significant differences in minutes per day between filter options for nonwearing time and time spent in sedentary, light, and moderate-to-vigorous physical activity (all P < 0.001), with more remarkable differences in the lower range of activity (sedentary and light activities). Mean counts per minute and steps per day were significantly higher using the low-frequency extension filter (P < 0.001). Most differences could be resolved using the correction factors. The observed differences are especially important when research is focusing on sedentary and light activities. In future studies, it is important to carefully evaluate the suitable filter option and to specify the filter choice in publications. The correction factors can be used to make data assessed using the low-frequency extension filter comparable to data assessed using the normal filter option.

  17. Sequential cloning of chromosomes

    SciTech Connect

    Lacks, S.A.

    1991-12-31

    A method for sequential cloning of chromosomal DNA and chromosomal DNA cloned by this method are disclosed. The method includes the selection of a target organism having a segment of chromosomal DNA to be sequentially cloned. A first DNA segment, having a first restriction enzyme site on either side. homologous to the chromosomal DNA to be sequentially cloned is isolated. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes.

  18. The First Human Cloned Embryo.

    ERIC Educational Resources Information Center

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  19. The First Human Cloned Embryo.

    ERIC Educational Resources Information Center

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  20. [Nuclear transfer and therapeutic cloning].

    PubMed

    Xu, Xiao-Ming; Lei, An-Min; Hua, Jin-Lian; Dou, Zhong-Ying

    2005-03-01

    Nuclear transfer and therapeutic cloning have widespread and attractive prospects in animal agriculture and biomedical applications. We reviewed that the quality of oocytes and nuclear reprogramming of somatic donor cells were the main reasons of the common abnormalities in cloned animals and the low efficiency of cloning and showed the problems and outlets in therapeutic cloning, such as some basic problems in nuclear transfer affected clinical applications of therapeutic cloning. Study on isolation and culture of nuclear transfer embryonic stem (ntES) cells and specific differentiation of ntES cells into important functional cells should be emphasized and could enhance the efficiency. Adult stem cells could help to cure some great diseases, but could not replace therapeutic cloning. Ethics also impeded the development of therapeutic cloning. It is necessary to improve many techniques and reinforce the research of some basic theories, then somatic nuclear transfer and therapeutic cloning may apply to agriculture reproduction and benefit to human life better.

  1. Cloning Components of Human Telomerase.

    DTIC Science & Technology

    1998-07-01

    nuclear factor NF90 homolog. (5 clones). RNA binding protein. Poorly understood. 3. FRG1 . Poorly understood. 4. DEK. Weak homology to Tetrahymena p95...least some of the clones for poorly understood genes (e.g. Hax-1, FRG1 , NF90, NF45, KIAA0098, KIAA0026, BAC397c4). Aim II. Functional Cloning of the

  2. Water relations of populus clones

    SciTech Connect

    Pallardy, S.G.; Kozlowski, T.T.

    1981-02-01

    Stomatal aperture and water balance in the field of eight Populus clones varying in growth rate were closely related to environmental factors and clonal differences were clearly expressed. Leaf water potential (psi) was influenced by solar radiation, leaf conductance, evaporative demand, and soil moisture content. The effects of soil moisture on psi were greatly modified by atmospheric conditions and stomatal conductance. Several slow-growing clones exhibited extended periods of psi below that of rapidly growing clones, despite high evaporative demand and the much greater transpiring surfaces of the fast-growing clones. Stomata of all clones responded to changes in light intensity and vapor pressure gradient (VPG). Pronounced stomatal sensitivity to VPG of two rapidly growing clones of common parentage, and the resultant capacity of these clones to moderate water deficits under high evaporative demand, were associated with drought resistance in one of the parents. Seasonal maximum leaf conductance was positively related to growth in several clones, suggesting that rapidly growing clones possess the capacity to carry on higher rates of gas exchange under favorable conditions. Analysis of changes in psi with changes in transpirational flux density (TFD) showed that for four clones, psi change per unit change in TFD decreased as TFD increased, indicating plant adaptation for prevention of damaging psi even at high TFD. More rapidly growing clones exhibited a larger initial rate of decline in psi with TFD, but reduced the rate of decline more than slow-growing clones as TFD increased. (Refs. 41).

  3. Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    PubMed Central

    Throop, Andrea L.; LaBaer, Joshua

    2015-01-01

    The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full-length cDNAs in species-specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site-specific recombination that is independent of the insert DNA sequence of interest, which differentiates this method from the classical restriction enzyme-based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression-ready clones using the most popular recombinational cloning technologies, including the commercially available Gateway® (Life Technologies) and In-Fusion® (Clontech) cloning technologies. PMID:25827088

  4. Three concepts of cloning in human beings.

    PubMed

    Cui, Ke-Hui

    2005-07-01

    Human cloning, organ cloning and tissue cloning are various types of cloning that occur at different levels with different methodologies. According to three standards of terminology for an embryo (fertilization through germ cells, development in the uterus and having the potential to produce a human life), tissue cloning and type I organ cloning will not produce an embryo. In contrast, human cloning and type II organ cloning will produce an embryo. Thus, only non-germinal tissue cloning and type I organ cloning are beyond the ethical question and will not change human beings as a species. Using cloned tissues to make new tissues or organs is promising for the future of medicine.

  5. Somatic c.34G>T KRAS mutation: a new prescreening test for MUTYH-associated polyposis?

    PubMed

    Aimé, Adeline; Coulet, Florence; Lefevre, Jeremie H; Colas, Chrystelle; Cervera, Pascale; Flejou, Jean-François; Lascols, Olivier; Soubrier, Florent; Parc, Yann

    2015-01-01

    We investigated the somatic c.34G>T KRAS transversion as a marker suggestive of MUTYH-associated polyposis (MAP). We compared 86 adenomas and 19 colorectal cancers (CRCs) of 30 MAP patients to 135 adenomas and five CRCs of 47 familial adenomatous polyposis (FAP) patients. The c.34G>T mutation was investigated by DNA sequencing. Secondly, the germline MUTYH gene sequence was analyzed in patients carrying c.34G>T in CRCs diagnosed between 2008 and 2012. The c.34G>T was present in 39.7% of MAP adenomas versus 1.6% of FAP adenomas (P < 0.01). Sensitivity and specificity for detecting MAP were 39.7% and 98%, respectively. Sensitivity increased with the number of adenomas tested (P = 0.039). KRAS exon 2 analysis was performed on 2239 CRC and 2.2% harbored the c.34G>T transversion. Among 28 carriers of the c.34G>T mutation, biallelic MUTYH mutations were detected in seven patients (25%). One patient did not have any polyp or family history and did not fulfill criteria for MUTYH testing. With high specificity, the c.34G>T mutation seems to be a useful and promising test for MAP. For polyposis, it may guide genetic testing toward APC or MUTYH. If routinely performed in CRC patients, it could help to diagnose MUTYH-mutation carriers, even when they don't fulfill genetic testing criteria. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Sayarim Infrasound Calibration Explosion provides first GT0 dataset for CTBTO

    NASA Astrophysics Data System (ADS)

    Gitterman, Yefim

    2010-05-01

    The large-scale calibration explosion of about 82 tons of HE explosives, assembled as a pyramid on the soft sediment surface, was successfully conducted by the Geophysical Institute of Israel at Sayarim Military Range on 26 August 2009. High-pressure values, measured in the range 200-600 m, were higher than predicted, whereas the created crater and seismic magnitude were smaller than expected for this on-surface charge. These results confirm that the used explosives, charge design and upward detonation provided the necessary explosion energy generation and partition: maximum of energy to the atmosphere and minimum to the ground. The high-pressure observations were utilized for estimation of the important Ground Truth parameter - TNT equivalent yield of about 0.1 kT (based on positive impulse in air-shock wave). Thus the Sayarim Explosion provided the first full GT0 source dataset for on-surface large-scale explosions, recorded by infrasound stations of International Monitoring System (IMS). Infrasound signals were well observed at distances up to 3,500 km, at numerous portable and permanent stations in Israel, Mediterranean countries and north-central Europe, including two IMS stations I26DE and I48TN and two portable arrays in Austria and Northern Italy deployed by the CTBTO team. Obtained records were used for analysis of infrasound signal propagation, source location and yield estimation, and comparison with GT0 source parameters.

  7. Subunit profiling and functional characteristics of acetylcholine receptors in GT1-7 cells.

    PubMed

    Arai, Yuki; Ishii, Hirotaka; Kobayashi, Makito; Ozawa, Hitoshi

    2017-03-01

    GnRH neurons form a final common pathway for the central regulation of reproduction. Although the involvement of acetylcholine in GnRH secretion has been reported, direct effects of acetylcholine and expression profiles of acetylcholine receptors (AChRs) still remain to be studied. Using immortalized GnRH neurons (GT1-7 cells), we analyzed molecular expression and functionality of AChRs. Expression of the mRNAs were identified in the order α7 > β2 = β1 ≧ α4 ≧ α5 = β4 = δ > α3 for nicotinic acetylcholine receptor (nAChR) subunits and m4 > m2 for muscarinic acetylcholine receptor (mAChR) subtypes. Furthermore, this study revealed that α7 nAChRs contributed to Ca(2+) influx and GnRH release and that m2 and m4 mAChRs inhibited forskolin-induced cAMP production and isobutylmethylxanthine-induced GnRH secretion. These findings demonstrate the molecular profiles of AChRs, which directly contribute to GnRH secretion in GT1-7 cells, and provide one possible regulatory action of acetylcholine in GnRH neurons.

  8. Structure–function relationships of membrane-associated GT-B glycosyltransferases†

    PubMed Central

    Albesa-Jové, David; Giganti, David; Jackson, Mary; Alzari, Pedro M; Guerin, Marcelo E

    2014-01-01

    Membrane-associated GT-B glycosyltransferases (GTs) comprise a large family of enzymes that catalyze the transfer of a sugar moiety from nucleotide-sugar donors to a wide range of membrane-associated acceptor substrates, mostly in the form of lipids and proteins. As a consequence, they generate a significant and diverse amount of glycoconjugates in biological membranes, which are particularly important in cell–cell, cell–matrix and host–pathogen recognition events. Membrane-associated GT-B enzymes display two “Rossmann-fold” domains separated by a deep cleft that includes the catalytic center. They associate permanently or temporarily to the phospholipid bilayer by a combination of hydrophobic and electrostatic interactions. They have the remarkable property to access both hydrophobic and hydrophilic substrates that reside within chemically distinct environments catalyzing their enzymatic transformations in an efficient manner. Here, we discuss the considerable progress that has been made in recent years in understanding the molecular mechanism that governs substrate and membrane recognition, and the impact of the conformational transitions undergone by these GTs during the catalytic cycle. PMID:24253765

  9. Designing Ground Antennas for Maximum G/T: Cassegrain or Gregorian?

    NASA Technical Reports Server (NTRS)

    Imbriale, William A.

    2005-01-01

    For optimum performance, a ground antenna system must maximize the ratio of received signal to the receiving system noise power, defined as the ratio of antenna gain to system-noise temperature (G/T). The total system noise temperature is the linear combination of the receiver noise temperature (including the feed system losses) and the antenna noise contribution. Hence, for very low noise cryogenic receiver systems, antenna noise-temperature properties are very significant contributors to G/T.It is well known that, for dual reflector systems designed for maximum gain, the gain performance of the antenna system is the same for both Cassegrain and Gregorian configurations. For a12-meter antenna designed to be part of the large array based Deep Space Network, a Cassegrain configuration designed for maximum G/T at X-band was 0.7 dB higher than the equivalent Gregorian configuration. This study demonstrates that, for maximum GIT, the dual shaped Cassegrain design is always better than the Gregorian.

  10. Endometrial cancer and somatic G>T KRAS transversion in patients with constitutional MUTYH biallelic mutations.

    PubMed

    Tricarico, Rossella; Bet, Paola; Ciambotti, Benedetta; Di Gregorio, Carmela; Gatteschi, Beatrice; Gismondi, Viviana; Toschi, Benedetta; Tonelli, Francesco; Varesco, Liliana; Genuardi, Maurizio

    2009-02-18

    MUTYH-associated polyposis (MAP) is an autosomal recessive condition predisposing to colorectal cancer, caused by constitutional biallelic mutations in the base excision repair (BER) gene MUTYH. Colorectal tumours from MAP patients display an excess of somatic G>T mutations in the APC and KRAS genes due to defective BER function. To date, few extracolonic manifestations have been observed in MAP patients, and the clinical spectrum of this condition is not yet fully established. Recently, one patient with a diagnosis of endometrial cancer and biallelic MUTYH mutations has been described. We here report on two additional unrelated MAP patients with biallelic MUTYH germline mutations who developed endometrioid endometrial carcinoma. The endometrial tumours were evaluated for PTEN, PIK3CA, KRAS, BRAF and CTNNB1 mutations. A G>T transversion at codon 12 of the KRAS gene was observed in one tumour. A single 1bp frameshift deletion of PTEN was observed in the same sample. Overall, these findings suggest that endometrial carcinoma is a phenotypic manifestations of MAP and that inefficient repair of oxidative damage can be involved in its pathogenesis.

  11. Mammalian cloning: advances and limitations.

    PubMed

    Solter, D

    2000-12-01

    For many years, researchers cloning mammals experienced little success, but recent advances have led to the successful cloning of several mammalian species. However, cloning by the transfer of nuclei from adult cells is still a hit-and-miss procedure, and it is not clear what technical and biological factors underlie this. Our understanding of the molecular basis of reprogramming remains extremely limited and affects experimental approaches towards increasing the success rate of cloning. Given the future practical benefits that cloning can offer, the time has come to address what should be done to resolve this problem.

  12. Sequential cloning of chromosomes

    DOEpatents

    Lacks, Sanford A.

    1995-07-18

    A method for sequential cloning of chromosomal DNA of a target organism is disclosed. A first DNA segment homologous to the chromosomal DNA to be sequentially cloned is isolated. The first segment has a first restriction enzyme site on either side. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism's chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction (class IIS) enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes.

  13. Sequential cloning of chromosomes

    DOEpatents

    Lacks, S.A.

    1995-07-18

    A method for sequential cloning of chromosomal DNA of a target organism is disclosed. A first DNA segment homologous to the chromosomal DNA to be sequentially cloned is isolated. The first segment has a first restriction enzyme site on either side. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction (class IIS) enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes. 9 figs.

  14. Ethical issues in livestock cloning.

    PubMed

    Thompson, P B

    1999-01-01

    Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity.

  15. Probabilistic cloning of equidistant states

    SciTech Connect

    Jimenez, O.; Roa, Luis; Delgado, A.

    2010-08-15

    We study the probabilistic cloning of equidistant states. These states are such that the inner product between them is a complex constant or its conjugate. Thereby, it is possible to study their cloning in a simple way. In particular, we are interested in the behavior of the cloning probability as a function of the phase of the overlap among the involved states. We show that for certain families of equidistant states Duan and Guo's cloning machine leads to cloning probabilities lower than the optimal unambiguous discrimination probability of equidistant states. We propose an alternative cloning machine whose cloning probability is higher than or equal to the optimal unambiguous discrimination probability for any family of equidistant states. Both machines achieve the same probability for equidistant states whose inner product is a positive real number.

  16. Microsatellite (GT)(n) repeats and SNPs in the von Willebrand factor gene promoter do not influence circulating von Willebrand factor levels under normal conditions.

    PubMed

    Daidone, Viviana; Cattini, Maria Grazia; Pontara, Elena; Sartorello, Francesca; Gallinaro, Lisa; Marotti, Alberto; Scaroni, Carla; Pagnan, Antonio; Casonato, Alessandra

    2009-02-01

    Von Willebrand factor (VWF) levels vary considerably in normal individuals, influenced by inherited and acquired modulators. ABO blood group is the major inherited determinant of VWF levels, but a role has also been attributed to the VWF gene promoter, haplotype 1 (-3268G/-2709C/-2661A/-2527G) being associated with higher VWF levels than haplotype 2 (-3268C/-2709T/-2661G/-2527A), and the polymorphic locus (GT)(n) modulating the shear stress-induced activation of the VWF promoter. We characterized the (GT)(n) of the VWF promoter in 394 healthy individuals and assessed whether its variable length influenced VWF levels in normal conditions. (GT)(n) proved highly polymorphic, with alleles from 15 to 24 repeats long. (GT)(21) and (GT)(19) were the most common variants (37.4% and 34.4%, respectively). Short GT repeats (15-19) segregated mainly with haplotype 1, long GT repeats (20-24) with haplotype 2 (p < 0.0001). The number of GT repeats did not correlate with VWF levels, nor did such levels correlate with haplotypes 1 and 2, considered alone or in association with the (GT)(n) locus. We conclude that (GT)(n) and -3268/-2709/-2661/-2527 loci are in strong linkage disequilibrium. This polymorphic region of the VWF promoter does not affect VWF levels under normal conditions, though it might represent an environmentally activable VWF regulation site.

  17. Comparison of ActiGraph GT3X+ and StepWatch Step Count Accuracy in Geriatric Rehabilitation Patients.

    PubMed

    Webber, Sandra C; St John, Philip D

    2016-07-01

    Activity monitors may not accurately detect steps in hospitalized older adults who walk slowly. We compared ActiGraph GT3X+ step counts (hip and ankle locations, default and low frequency extension [LFE] analyses) to the StepWatch monitor (ankle) during a hallway walk in 38 geriatric rehabilitation patients (83.2 ± 7.1 years of age, 0.4 ± 0.2 m/s gait speed). Absolute percent error values were low (<3%) and did not differ for the StepWatch and the GT3X+ (ankle, LFE); however, error values were high (19-97%) when the GT3X+ was worn at the hip and/ or analyzed with the default filter. Although these finding suggest the GT3X+ (ankle, LFE) functions as well as the StepWatch in detecting steps during walking in older adults with slow gait speeds, further research is needed to determine whether the GT3X+ is also able to disregard other body movements (e.g., fidgeting) that occur when full day monitoring is utilized.

  18. Gene cloning and protein expression of γ-glutamyltranspeptidases from Thermus thermophilus and Deinococcus radiodurans: comparison of molecular and structural properties with mesophilic counterparts.

    PubMed

    Castellano, Immacolata; Di Salle, Anna; Merlino, Antonello; Rossi, Mosè; La Cara, Francesco

    2011-03-01

    γ-Glutamyltranspeptidase (γ-GT) is an ubiquitous enzyme that catalyzes the hydrolysis of γ-glutamyl bonds in glutathione and glutamine and the transfer of the released γ-glutamyl group to amino acids or short peptides. γ-GTs from extremophiles, bacteria adapted to live in hostile environments, were selected as model systems to study the molecular underpinnings of their adaptation to extreme conditions and to find out special properties of potential biotechnological interest. Here, we report the cloning, expression and purification of two members of γ-GT family from two different extremophilic species, Thermus thermophilus (TtGT) and Deinococcus radiodurans (DrGT); the first is an aerobic eubacterium, growing at high temperatures (50-82°C), the second is a polyextremophile, as it tolerates radiations, cold, dehydration, vacuum, and acid. TtGT and DrGT were both synthesized as precursor proteins of 59-60 kDa, undergoing an intramolecular auto-cleavage to yield two subunits of 40 and 19-20 kDa, respectively. However, like the γ-GT from Geobacillus thermodenitrificans, but differently from the other characterized bacterial and eukaryotic γ-GTs, the two new extremophilic enzymes displayed γ-glutamyl hydrolase, but not transpeptidase activity in the 37-50°C temperature range, pH 8.0. The comparison of sequences and structural models of these two proteins with experimental-determined structures of other known mesophilic γ-GTs suggests that the extremophilic members of this protein family have found a common strategy to adapt to different hostile environments. Moreover, a phylogenetic analysis suggests that γ-GTs displaying only γ-glutamyl hydrolase activity could represent the progenitors of the bacterial and eukaryotic counterparts.

  19. Ground Reaction Force Estimates from ActiGraph GT3X+ Hip Accelerations

    PubMed Central

    Neugebauer, Jennifer M.; Collins, Kelsey H.; Hawkins, David A.

    2014-01-01

    Simple methods to quantify ground reaction forces (GRFs) outside a laboratory setting are needed to understand daily loading sustained by the body. Here, we present methods to estimate peak vertical GRF (pGRFvert) and peak braking GRF (pGRFbrake) in adults using raw hip activity monitor (AM) acceleration data. The purpose of this study was to develop a statistically based model to estimate pGRFvert and pGRFbrake during walking and running from ActiGraph GT3X+ AM acceleration data. 19 males and 20 females (age 21.2±1.3 years, height 1.73±0.12 m, mass 67.6±11.5 kg) wore an ActiGraph GT3X+ AM over their right hip. Six walking and six running trials (0.95–2.19 and 2.20–4.10 m/s, respectively) were completed. Average of the peak vertical and anterior/posterior AM acceleration (ACCvert and ACCbrake, respectively) and pGRFvert and pGRFbrake during the stance phase of gait were determined. Thirty randomly selected subjects served as the training dataset to develop generalized equations to predict pGRFvert and pGRFbrake. Using a holdout approach, the remaining 9 subjects were used to test the accuracy of the models. Generalized equations to predict pGRFvert and pGRFbrake included ACCvert and ACCbrake, respectively, mass, type of locomotion (walk or run), and type of locomotion acceleration interaction. The average absolute percent differences between actual and predicted pGRFvert and pGRFbrake were 8.3% and 17.8%, respectively, when the models were applied to the test dataset. Repeated measures generalized regression equations were developed to predict pGRFvert and pGRFbrake from ActiGraph GT3X+ AM acceleration for young adults walking and running. These equations provide a means to estimate GRFs without a force plate. PMID:24914946

  20. To clone or not to clone--a Jewish perspective.

    PubMed Central

    Lipschutz, J H

    1999-01-01

    Many new reproductive methods such as artificial insemination, in vitro fertilisation, freezing of human embryos, and surrogate motherhood were at first widely condemned but are now seen in Western society as not just ethically and morally acceptable, but beneficial in that they allow otherwise infertile couples to have children. The idea of human cloning was also quickly condemned but debate is now emerging. This article examines cloning from a Jewish perspective and finds evidence to support the view that there is nothing inherently wrong with the idea of human cloning. A hypothesis is also advanced suggesting that even if a body was cloned, the brain, which is the essence of humanity, would remain unique. This author suggests that the debate should be changed from "Is cloning wrong?" to "When is cloning wrong?". PMID:10226913

  1. Ethical issues in animal cloning.

    PubMed

    Fiester, Autumn

    2005-01-01

    The issue of human reproductive cloning has recently received a great deal attention in public discourse. Bioethicists, policy makers, and the media have been quick to identify the key ethical issues involved in human reproductive cloning and to argue, almost unanimously, for an international ban on such attempts. Meanwhile, scientists have proceeded with extensive research agendas in the cloning of animals. Despite this research, there has been little public discussion of the ethical issues raised by animal cloning projects. Polling data show that the public is decidedly against the cloning of animals. To understand the public's reaction and fill the void of reasoned debate about the issue, we need to review the possible objections to animal cloning and assess the merits of the anti-animal cloning stance. Some objections to animal cloning (e.g., the impact of cloning on the population of unwanted animals) can be easily addressed, while others (e.g., the health of cloned animals) require more serious attention by the public and policy makers.

  2. Isolation of cDNA clones for differentially expressed genes of the human parasite Schistosoma mansoni.

    PubMed Central

    Davis, A H; Blanton, R; Rottman, F; Maurer, R; Mahmoud, A

    1986-01-01

    Little is known about the mechanisms that control transformations during the life cycle of Schistosoma mansoni. To enable isolation of DNA sequences encoding developmentally regulated antigens a cDNA expression library in the vector lambda gt11 amp3 was constructed from adult mRNA and immunologically screened with sera from infected individuals. We report here on the properties of three recombinant clones that derive from developmentally regulated genes. Clone 10-3 encoded a beta-galactosidase fusion protein present in high abundance in infected Escherichia coli. Clones 7-2 and 8-2 also produced immunologically recognized proteins; however, the peptides did not appear to be beta-galactosidase fusion proteins. The expression of mRNAs hybridizing to these cDNAs was examined in the different stages of the parasite life cycle. Messenger RNA corresponding to clone 10-3, approximately equal to 1000 bases in length, was present in higher abundance in male worms than in females but was not detected in schistosome eggs. A 900-base mRNA hybridizing to clone 7-2 was observed in adult worms and eggs. Both clone 10-3 and clone 7-2 hybridized to smaller mRNAs in cercariae and freshly transformed schistosomula than in adult worms. Clone 8-2 contained tandem cDNA inserts. One cDNA hybridized to a 1700-base mRNA present in all stages, while the second hybridized to an 800-base mRNA specific to adult female worms. Images PMID:3461448

  3. To clone or not to clone--whither the law?

    PubMed

    Lupton, M L

    1999-01-01

    The cloning of Dolly the lamb from adult cells by scientists at the Roslin Laboratories near Edinburgh in February 1997 has startled the world because it now opens the way to clone adult human beings. The reaction to Ian Wilmut's breakthrough has been instant and largely negative. Bills were rushed into both the US Senate and House of Representatives aimed at banning the cloning of human beings. Human cloning is premature at this stage, but there are many positive spin-offs of cloning in the field of genetic engineering, such as the production of human proteins such as blood clotting factors which aid in healing wounds. Progress by means of cloning can also be made into devising a cure for Parkinson's Disease amongst others. No lesser ethicist than John C. Fletcher of the University of Virginia foresees circumstances in which human cloning is acceptable e.g. to enable a couple to replace a dying child, to enable a couple, one of whom is infertile, to clone a child from either partner. Extensive regulation of cloning by the law is inevitable but, in doing so, the legislation should be careful not to outlaw research in this area which could be beneficial to mankind.

  4. Correlation between MTP -493G>T polymorphism and non-alcoholic fatty liver disease risk: a meta-analysis.

    PubMed

    Li, L; Wang, S J; Shi, K; Chen, D; Jia, H; Zhu, J

    2014-12-04

    Several studies have found that microsomal transfer protein (MTP) may be important in the development and progression of non-alcoholic fatty liver disease (NAFLD). In this meta-analysis, we evaluated the relationships between a common polymorphism (-493G>T, rs1800591 G>T) in the MTP gene and NAFLD risk. The PubMed, CISCOM, CINAHL, Web of Science, Google Scholar, EBSCO, Cochrane Library, and CBM databases were searched for relevant articles published before October 1, 2013 without any language restrictions. Meta-analysis was conducted using the STATA 12.0 software. Crude odds ratios (ORs) with 95% confidence intervals (95%CIs) were calculated. Eleven case-control studies were included in this meta-analysis. A total of 636 NAFLD patients and 918 healthy control subjects were examined in this meta-analysis. Our results indicate that the MTP -493G/T polymorphism increases the risk of NAFLD (G allele vs T allele: OR = 1.39, 95%CI = 1.17-1.65, P < 0.001; GG + GT vs TT: OR = 1.46, 95%CI = 1.02-2.09, P = 0.038, respectively). Subgroup analyses indicated that the MTP -493G/T polymorphism was associated with an increased risk of NAFLD in population-based, hospital-based, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and large sample-size subgroups under the allele and dominant models (all P < 0.05). However, we found no association between non-PCR-RFLP polymorphism and small sample-size subgroups (all P > 0.05). Our findings indicate that the MTP -493G/ T polymorphism may contribute to the development of NAFLD. Thus, the MTP -493G/T polymorphism may be a biomarker for the early detection of NAFLD.

  5. Heterogeneity of rat tropoelastin mRNA revealed by cDNA cloning

    SciTech Connect

    Pierce, R.A.; Deak, S.B.; Stolle, C.A.; Boyd, C.D. )

    1990-10-01

    A {lambda}gt11 library constructed from poly(A{plus}) RNA isolated from aortic tissue of neonatal rats was screened for rat tropoelastin cDNAs. The first, screen, utilizing a human tropoelastin cDNA clone, provided rat tropoelastin cDNAs spanning 2.3 kb of carboxy-terminal coding sequence and extended into the 3{prime}-untranslated region. A subsequent screen using a 5{prime} rat tropoelastin cDNA clone yielded clones extending into the amino-terminal signal sequence coding region. Sequence analysis of these clones has provided the complete derived amino acid sequence of rat tropoelastin and allowed alignment and comparison with published bovine cDNA sequence. While the overall structure of rat tropoelastin is similar to bovine sequence, numerous substitutions, deletions, and insertions demonstrated considerable heterogeneity between species. In particular, the pentapeptide repeat VPGVG, characteristic of all tropoelastins analyzed to date, is replaced in rat tropoelastin by a repeating pentapeptide, IPGVG. The hexapeptide repeat VGVAPG, the bovine elastin receptor binding peptide, is not encoded by rat tropoelastin cDNAs. Variations in coding sequence between rat tropoelastin CDNA clones were also found which may represent mRNA heterogeneity produced by alternative splicing of the rat tropoelastin pre-mRNA.

  6. Biocompatibility studies of natural rubber latex from different tree clones and collection methods.

    PubMed

    Floriano, Juliana Ferreira; da Mota, Lígia Souza Lima Silveira; Furtado, Edson Luiz; Rossetto, Victor José Vieira; Graeff, Carlos F O

    2014-02-01

    Natural rubber latex (NRL) has several features that make it an excellent biomaterial to promote the growth and repair of tissues, skin and bones. Most of the research with NRL membranes uses a mixture of different clones and chemical preservatives in the collection process. In this study, we compared five clones that produce NRL, seeking to identify their differences in biocompatibility. The clones studied were RRIM 600, PB 235, GT1, PR 255 and IAN 873 commonly found in plantations in Brazil. We did also study the effect of ammonia used during latex collection. NRL membranes were prepared aseptically and sterilized. In the in vitro tests, the membranes remained in direct contact with mouse fibroblasts cells for three periods, 24, 48 and 72 h. In the in vivo tests, the membranes were implanted subcutaneously in rabbits. The results indicated the biocompatibility of the membranes obtained from all clones. Membranes from the clones RRIM 600 and IAN 873 induced greater cell proliferation, suggesting greater bioactivity. It was found that the membranes made from latex that was in contact with ammonia during collection, showed cytotoxic and genotoxic effects in cultures, as well as necrosis, and increased inflammatory cells in the rabbit's tissues close to the implant.

  7. Lessons learned from cloning dogs.

    PubMed

    Kim, M J; Oh, H J; Kim, G A; Park, J E; Park, E J; Jang, G; Ra, J C; Kang, S K; Lee, B C

    2012-08-01

    The aim of this article is to review dog cloning research and to suggest its applications based on a discussion about the normality of cloned dogs. Somatic cell nuclear transfer was successfully used for production of viable cloned puppies despite limited understanding of in vitro dog embryo production. Cloned dogs have similar growth characteristics to those born from natural fertilization, with no evidence of serious adverse effects. The offspring of cloned dogs also have similar growth performance and health to those of naturally bred puppies. Therefore, cloning in domestic dogs can be applied as an assisted reproductive technique to conserve endangered species, to treat sterile canids or aged dogs, to improve reproductive performance of valuable individuals and to generate disease model animals. © 2012 Blackwell Verlag GmbH.

  8. Therapeutic cloning and reproductive liberty.

    PubMed

    Sparrow, Robert

    2009-04-01

    Concern for "reproductive liberty" suggests that decisions about embryos should normally be made by the persons who would be the genetic parents of the child that would be brought into existence if the embryo were brought to term. Therapeutic cloning would involve creating and destroying an embryo, which, if brought to term, would be the offspring of the genetic parents of the person undergoing therapy. I argue that central arguments in debates about parenthood and genetics therefore suggest that therapeutic cloning would be prima facie unethical unless it occurred with the consent of the parents of the person being cloned. Alternatively, if therapeutic cloning is thought to be legitimate, this undermines the case for some uses of reproductive cloning by implying that the genetic relation it establishes between clones and DNA donors does not carry the same moral weight as it does in cases of normal reproduction.

  9. Equine cloning: applications and outcomes.

    PubMed

    Vanderwall, Dirk K; Woods, Gordon L; Roser, Janet F; Schlafer, Donald H; Sellon, Debra C; Tester, David F; White, Kenneth L

    2006-01-01

    Cloning is one of several new assisted reproductive techniques being developed for clinical use in the equine industry. Potential uses of equine cloning include: (1) the preservation of genetics from individual animals that would otherwise not be able to reproduce, such as geldings; (2) the preservation of genetic material of endangered and/or exotic species, such as the Mongolian wild horse (Przewalski's horse); and (3) because of the companion animal role that horses fill for some individuals, it is likely that some horse owners will have individual animals cloned for emotional fulfillment. Although equine cloning has been successful, like other species, it remains a very inefficient process (<3% success). In most species, the inefficiency of cloning results from a high incidence of embryonic, fetal and/or placental developmental abnormalities that contribute to extremely high rates of embryonic loss, abortion and stillbirths throughout gestation and compromised neonatal health after birth. The present review describes some of the ultrasonographic, endocrinological and histopathological characteristics of successful (produced viable offspring) and unsuccessful (resulted in pregnancy failure) cloned equine (mule and horse) pregnancies we have produced. A total of 21 cloned mule pregnancies were established using fetal fibroblast cells, whereas a total of seven cloned horse pregnancies were established using adult cumulus cells. Three of the cloned mule conceptuses were carried to term, resulting in the birth of three healthy clones. This information adds to an accumulating body of knowledge about the outcome of cloned equine pregnancies, which will help to establish when, and perhaps why, many cloned equine pregnancies fail.

  10. Guide to molecular cloning techniques

    SciTech Connect

    Berger, S.L.; Kimmel, A.R.

    1987-01-01

    This book includes the following selections: requirements for a molecular biology laboratory; general methods for isolating and characterizing nucleic acids; enzymatic techniques and recombinant DNA technology; restriction enzymes; growth and maintenance of bacteria; genetic cloning, preparation and characterization of RNA; preparation of cDNA and the generation of cDNA libraries; selections of clones from libraries; and identification and characterization of specific clones.

  11. Therapeutic cloning: The ethical limits

    SciTech Connect

    Whittaker, Peter A. . E-mail: p.whittaker@lancaster.ac.uk

    2005-09-01

    A brief outline of stem cells, stem cell therapy and therapeutic cloning is given. The position of therapeutic cloning with regard to other embryonic manipulations - IVF-based reproduction, embryonic stem formation from IVF embryos and reproductive cloning - is indicated. The main ethically challenging stages in therapeutic cloning are considered to be the nuclear transfer process including the source of eggs for this and the destruction of an embryo to provide stem cells for therapeutic use. The extremely polarised nature of the debate regarding the status of an early human embryo is noted, and some potential alternative strategies for preparing immunocompatible pluripotent stem cells are indicated.

  12. Cloning to reproduce desired genotypes.

    PubMed

    Westhusin, M E; Long, C R; Shin, T; Hill, J R; Looney, C R; Pryor, J H; Piedrahita, J A

    2001-01-01

    Cloned sheep, cattle, goats, pigs and mice have now been produced using somatic cells for nuclear transplantation. Animal cloning is still very inefficient with on average less than 10% of the cloned embryos transferred resulting in a live offspring. However successful cloning of a variety of different species and by a number of different laboratory groups has generated tremendous interest in reproducing desired genotypes. Some of these specific genotypes represent animal cell lines that have been genetically modified. In other cases there is a significant demand for cloning animals characterized by their inherent genetic value, for example prize livestock, household pets and rare or endangered species. A number of different variables may influence the ability to reproduce a specific genotype by cloning. These include species, source of recipient ova, cell type of nuclei donor, treatment of donor cells prior to nuclear transfer, and the techniques employed for nuclear transfer. At present, there is no solid evidence that suggests cloning will be limited to only a few specific animals, and in fact, most data collected to date suggests cloning will be applicable to a wide variety of different animals. The ability to reproduce any desired genotype by cloning will ultimately depend on the amount of time and resources invested in research.

  13. Human cloning and child welfare.

    PubMed Central

    Burley, J; Harris, J

    1999-01-01

    In this paper we discuss an objection to human cloning which appeals to the welfare of the child. This objection varies according to the sort of harm it is expected the clone will suffer. The three formulations of it that we will consider are: 1. Clones will be harmed by the fearful or prejudicial attitudes people may have about or towards them (H1); 2. Clones will be harmed by the demands and expectations of parents or genotype donors (H2); 3. Clones will be harmed by their own awareness of their origins, for example the knowledge that the genetic donor is a stranger (H3). We will show why these three versions of the child welfare objection do not necessarily supply compelling reasons to ban human reproductive cloning. The claim that we will develop and defend in the course of our discussion is that even if it is the case that a cloned child will suffer harms of the type H1-H3, it is none the less permissible to conceive by cloning so long as these cloning-induced welfare deficits are not such as to blight the existence of the resultant child, whoever this may be. PMID:10226914

  14. Cloning and characterization of the aroA gene from Mycobacterium tuberculosis.

    PubMed Central

    Garbe, T; Jones, C; Charles, I; Dougan, G; Young, D

    1990-01-01

    The aroA gene from Mycobacterium tuberculosis has been cloned by complementation of an aroA mutant of Escherichia coli after lysogenization with a recombinant DNA library in the lambda gt11 vector. Detailed characterization of the M. tuberculosis aroA gene by nucleotide sequencing and by immunochemical analysis of the expressed product indicates that it encodes a 5-enolpyruvylshikimate-3-phosphate synthase that is structurally related to analogous enzymes from other bacterial, fungal, and plant sources. The potential use of the cloned gene in construction of genetically defined mutant strains of M. tuberculosis by gene replacement is proposed as a novel approach to the rational attenuation of mycobacterial pathogens and the possible development of new antimycobacterial vaccines. Images PMID:2123856

  15. NaCl stress induces CsSAMs gene expression in Cucumis sativus by mediating the binding of CsGT-3b to the GT-1 element within the CsSAMs promoter.

    PubMed

    Wang, Li-Wei; He, Mei-Wen; Guo, Shi-Rong; Zhong, Min; Shu, Sheng; Sun, Jin

    2017-05-01

    The CsSAMs promoter is a salt-stress-inducible promoter containing three GT-1 elements that are sufficient for the salt-stress response. The transcription factor CsGT-3b was found to bind to the GT-1 element. The S-adenosyl-L-methionine synthase (SAMs) gene is among the functional genes induced during environmental stress. However, little is known about the regulatory mechanism and upstream regulators of this salt-inducible gene in cucumber plants. Thus, it is necessary to understand the characteristics of the SAMs gene by analyzing its promoter and transcription factors. In this study, we isolated and functionally analyzed a 1743-bp flanking fragment of the CsSAMs gene from Cucumis sativus. To examine promoter activity, the full-length promoter, as well as different promoter fragments, were fused to the β-glucuronidase (GUS) reporter gene and introduced into the tobacco genome. The full-length promoter displayed maximal promoter activity, whereas the P4 promoter, containing 321 bp of upstream sequence, showed no basal promoter activity. In addition, the CsSAMs promoter exhibited stress-inducible regulation rather than tissue-specific activity in transgenic tobacco. Histochemical analysis revealed strong GUS staining in leaves, stems, and roots, especially in the veins of leaves, the vascular bundle of stems, and root tip zones following NaCl stress. A transient expression assay confirmed that the 242-bp region (-1743 to -1500) was sufficient for the NaCl-stress response. Yeast one-hybrid assays further revealed interaction between the NaCl-response protein CsGT-3b and the GT-1 (GAAAAA) element within the 242-bp region. Taken together, we revealed the presence of four salt-stress-responsive elements (GT-1 cis-elements) in the CsSAMs promoter and identified a transcription factor, CsGT-3b, that specifically binds to this sequence. These results might help us better understand the intricate regulatory network of the cucumber SAMs gene.

  16. Susceptibility of GT1-7 cells to mouse-passaged field scrapie isolates with a long incubation.

    PubMed

    Miyazawa, Kohtaro; Okada, Hiroyuki; Iwamaru, Yoshifumi; Masujin, Kentaro; Yokoyama, Takashi

    2014-01-01

    A typical feature of scrapie in sheep and goats is the accumulation of disease-associated prion protein. Scrapie consists of many strains with different biological properties. Nine natural sheep scrapie cases were transmitted to wild-type mice and mouse-passaged isolates were classified into 2 types based on incubation time: short and long. These 2 types displayed a distinct difference in their pathology. We attempted to transmit these mouse-passaged isolates to 2 murine cell lines (GT1-7 and L929) to compare their properties. All of the isolates were transmitted to L929 cells. However, only mouse-passaged field isolates with a long incubation time were transmitted to GT1-7 cells. This specific susceptibility of GT1-7 cells was also confirmed with a primary-passaged isolate that was not completely adapted to the new host species. Characterization of the mechanisms of the specific susceptibility of GT1-7 cells to isolates with a long incubation time may lead to a greater understanding of the differences among prion strains.

  17. The evaluation of MCI, MI, PMI and GT on both genders with different age and dental status.

    PubMed

    Bozdag, G; Sener, S

    2015-01-01

    The aim of this study was to measure the mandibular cortical index (MCI), mental index (MI), panoramic mandibular index (PMI) and cortical bone thickness in the zone of the gonial angle (GT) in panoramic radiographies from a large sample of males and females and to determine how they relate to patients' age, gender and dental status. 910 panoramic radiographs were obtained and grouped into age, dental status and gender. The MCI, MI, PMI and GT were analysed. Remarkable differences were observed for MCI and GT regarding gender, age groups and dental status on both sides (p < 0.05). While age and dental status had an effect on the MI and PMI in females, dental status had an effect on the MI and PMI in males (p < 0.05). Also, gender had an effect on the MI and PMI (p < 0.05). The effects of age and tooth loss are different in females and males. In females, the harmful effects of tooth loss and age are more prominent according to the PMI and MI measurements. The effects of age and tooth loss in the GT and MCI measurements are similar, and these indices can be accepted as more reliable in studies including both genders.

  18. The Arabidopsis Family GT43 Glycosyltransferases Form Two Functionally Nonredundant Groups Essential for the Elongation of Glucuronoxylan Backbone

    EPA Science Inventory

    There exist four members of family GT43 glycosyltransferases in the Arabidopsis (Arabidopsis thaliana) genome, and mutations of two of them, IRX9 and IRX14, have previously been shown to cause a defect in glucuronoxylan (GX) biosynthesis. However, it is currently unknown whether ...

  19. Novel expression hosts for complex secondary metabolite megasynthetases: Production of myxochromide in the thermopilic isolate Corallococcus macrosporus GT-2

    PubMed Central

    Perlova, Olena; Gerth, Klaus; Kuhlmann, Silvia; Zhang, Youming; Müller, Rolf

    2009-01-01

    Although many secondary metabolites with diverse biological activities have been isolated from myxobacteria, most strains of these biotechnologically important gliding prokaryotes remain difficult to handle genetically. In this study we describe the new fast growing myxobacterial thermophilic isolate GT-2 as a heterologous host for the expression of natural product biosynthetic pathways isolated from other myxobacteria. According to the results of sequence analysis of the 16S rDNA, this moderately thermophilic isolate is closely related to Corallococcus macrosporus and was therefore named C. macrosporus GT-2. Fast growth of moderately thermophilic strains results in shorter fermentation and generation times, aspects which are of significant interest for molecular biological work as well as production of secondary metabolites. Development of a genetic manipulation system allowed the introduction of the complete myxochromide biosynthetic gene cluster, located on a transposable fragment, into the chromosome of GT-2. Genetic engineering of the biosynthetic gene cluster by promoter exchange leads to much higher production of myxochromides in the heterologous host C. macrosporus GT-2 in comparison to the original producer Stigmatella aurantiaca and to the previously described heterologous host Pseudomonas putida (600 mg/L versus 8 mg/L and 40 mg/L, respectively). PMID:19126236

  20. DNMT3B 579G>T promoter polymorphism and the risk for idiopathic thrombocytopenic purpura in a Chinese population.

    PubMed

    Zhao, Haifeng; Du, Weiting; Gu, Dongsheng; Wang, Donghai; Xue, Feng; Ge, Jing; Sui, Tao; Yang, Renchi

    2009-01-01

    Epigenetics may influence the expression of numerous genes, which might contribute to autoimmune diseases. DNA methylation is mediated by DNA methyltransferases, especially DNA methyltransferase 3B (DNMT3B). Polymorphisms of the DNMT3B gene may influence DNMT3B activity on DNA methylation and increase the susceptibility to several diseases. The current study investigated the association between DNMT3B 579G>T and the risk for idiopathic thrombocytopenic purpura (ITP). The DNMT3B 579G>T polymorphisms were analyzed by PCR-RFLP. There was no significant difference in genotype and allele distribution between the ITP patient and the controls (p = 0.722 and 0.667, respectively). Similar results were observed between the 2 groups when stratified by age and disease course, including acute in childhood, chronic in childhood, acute in adult and chronic in adult. Importantly, this study showed a statistical difference in the distribution of SNP of DNMT3B between Chinese and Koreans or Americans. It is shown that the SNP of DNMT3B 579G>T may not be used on its own as a marker to predict the susceptibility to ITP in a Chinese population and that DNMT3B 579G>T promoter SNP varies from one ethnic population to another.

  1. Validation of the Actigraph GT3X and ActivPAL Accelerometers for the Assessment of Sedentary Behavior

    ERIC Educational Resources Information Center

    Kim, Youngdeok; Barry, Vaughn W.; Kang, Minsoo

    2015-01-01

    This study examined (a) the validity of two accelerometers (ActiGraph GT3X [ActiGraph LLC, Pensacola, FL, USA] and activPAL [PAL Technologies Ltd., Glasgow, Scotland]) for the assessment of sedentary behavior; and (b) the variations in assessment accuracy by setting minimum sedentary bout durations against a proxy for direct observation using an…

  2. Genetic mapping and QTL analysis of disease resistance traits in peanut population Tifrunner x GT-C20

    USDA-ARS?s Scientific Manuscript database

    A genetic map of peanut (Arachis hypogaea L.) with 426 SSR markers was constructed using a population of 162 recombinant inbred lines (RILs) from a cross between ‘Tifrunner’ and ‘GT-C20’. Linkage groups (LGs) were assigned to chromosomes using published peanut reference maps. The total length of the...

  3. GT microsatellite repeats in the heme oxygenase-1 gene promoter associated with abdominal aortic aneurysm in Croatian patients.

    PubMed

    Gregorek, Andrea Crkvenac; Gornik, Kristina Crkvenac; Polancec, Darija Stupin; Dabelic, Sanja

    2013-06-01

    Abdominal aortic aneurysm (AAA) is a complex genetic disorder caused by the interplay of genetic and environmental risk factors. The number of (GT)(n) repeats in the heme oxygenase-1 (HO-1) gene promoter modulates transcription of this enzyme, which might have anti-inflammatory, antioxidant, antiapoptotic, and antiproliferative effect. The distribution of alleles and genotypes in Croatian individuals genotyped for the (GT)(n) HO-1 polymorphism was similar to that in other European populations. Frequency of the short (S) alleles (GT < 25) was higher in AAA patients (41.9%) than in non-AAA individuals (28.2%, p = 0.0026) because there were more SL heterozygotes among the AAA patients. The SL genotype appeared to increase the risk for AAA, but the increase was not statistically significant after adjustment for age, sex, smoking, hypertension, and hyperlipidemia (OR = 1.53, 95% CI 0.90-3.09, p = 0.062). These findings contradict those of the only other study performed so far on the association of (GT)(n) HO-1 polymorphism and AAA.

  4. Validation of the Actigraph GT3X and ActivPAL Accelerometers for the Assessment of Sedentary Behavior

    ERIC Educational Resources Information Center

    Kim, Youngdeok; Barry, Vaughn W.; Kang, Minsoo

    2015-01-01

    This study examined (a) the validity of two accelerometers (ActiGraph GT3X [ActiGraph LLC, Pensacola, FL, USA] and activPAL [PAL Technologies Ltd., Glasgow, Scotland]) for the assessment of sedentary behavior; and (b) the variations in assessment accuracy by setting minimum sedentary bout durations against a proxy for direct observation using an…

  5. MTP -493G/T gene polymorphism is associated with steatosis in hepatitis C-infected patients.

    PubMed

    Siqueira, E R F; Oliveira, C P M S; Correa-Giannella, M L; Stefano, J T; Cavaleiro, A M; Fortes, M A H Z; Muniz, M T C; Silva, F S; Pereira, L M M B; Carrilho, F J

    2012-01-01

    The reduction of hepatic microsomal transfer protein (MTP) activity results in fatty liver, worsening hepatic steatosis and fibrosis in chronic hepatitis C (CHC). The G allele of the MTP gene promoter, -493G/T, has been associated with lower transcriptional activity than the T allele. We investigated this association with metabolic and histological variables in patients with CHC. A total of 174 untreated patients with CHC were genotyped for MTP -493G/T by direct sequencing using PCR. All patients were negative for markers of Wilson's disease, hemochromatosis and autoimmune diseases and had current and past daily alcohol intake lower than 100 g/week. The sample distribution was in Hardy-Weinberg equilibrium. Among subjects with genotype 1, 56.8% of the patients with fibrosis grade 3+4 presented at least one G allele versus 34.3% of the patients with fibrosis grade 1+2 (OR = 1.8; 95%CI = 1.3-2.3). Logistic regression analysis with steatosis as the dependent variable identified genotypes GG+GT as independent protective factors against steatosis (OR = 0.4, 95%CI = 0.2-0.8; P = 0.01). The results suggest that the presence of the G allele of MTP -493G/T associated with lower hepatic MTP expression protects against steatosis in our CHC patients.

  6. The Arabidopsis Family GT43 Glycosyltransferases Form Two Functionally Nonredundant Groups Essential for the Elongation of Glucuronoxylan Backbone

    EPA Science Inventory

    There exist four members of family GT43 glycosyltransferases in the Arabidopsis (Arabidopsis thaliana) genome, and mutations of two of them, IRX9 and IRX14, have previously been shown to cause a defect in glucuronoxylan (GX) biosynthesis. However, it is currently unknown whether ...

  7. CATO: The Clone Alignment Tool

    PubMed Central

    Henstock, Peter V.; LaPan, Peter

    2016-01-01

    High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1) a top-level summary of the top candidate sequences aligned to each reference sequence, 2) a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3) a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow. PMID:27459605

  8. CATO: The Clone Alignment Tool.

    PubMed

    Henstock, Peter V; LaPan, Peter

    2016-01-01

    High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1) a top-level summary of the top candidate sequences aligned to each reference sequence, 2) a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3) a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow.

  9. [The discrete horror of cloning].

    PubMed

    Guibourg, Ricardo A

    2009-01-01

    The author raises the topic of cloning after the decision of the Argentine government, which concerned for the "dignity of the human person", passed a decree of need and urgency, No. 200/97 (Annex), prohibiting cloning experiments with human beings. Therefore, considering that the topic is so terribly urgent and necessary, the author feels it is timely to consider it.

  10. Integration of a wave rotor to an ultra-micro gas turbine (UmuGT)

    NASA Astrophysics Data System (ADS)

    Iancu, Florin

    2005-12-01

    Wave rotor technology has shown a significant potential for performance improvement of thermodynamic cycles. The wave rotor is an unsteady flow machine that utilizes shock waves to transfer energy from a high energy fluid to a low energy fluid, increasing both the temperature and the pressure of the low energy fluid. Used initially as a high pressure stage for a gas turbine locomotive engine, the wave rotor was commercialized only as a supercharging device for internal combustion engines, but recently there is a stronger research effort on implementing wave rotors as topping units or pressure gain combustors for gas turbines. At the same time, Ultra Micro Gas Turbines (UmuGT) are expected to be a next generation of power source for applications from propulsion to power generation, from aerospace industry to electronic industry. Starting in 1995, with the MIT "Micro Gas Turbine" project, the mechanical engineering research world has explored more and more the idea of "Power MEMS". Microfabricated turbomachinery like turbines, compressors, pumps, but also electric generators, heat exchangers, internal combustion engines and rocket engines have been on the focus list of researchers for the past 10 years. The reason is simple: the output power is proportional to the mass flow rate of the working fluid through the engine, or the cross-sectional area while the mass or volume of the engine is proportional to the cube of the characteristic length, thus the power density tends to increase at small scales (Power/Mass=L -1). This is the so-called "cube square law". This work investigates the possibilities of incorporating a wave rotor to an UmuGT and discusses the advantages of wave rotor as topping units for gas turbines, especially at microscale. Based on documented wave rotor efficiencies at larger scale and subsidized by both, a gasdynamic model that includes wall friction, and a CFD model, the wave rotor compression efficiency at microfabrication scale could be estimated

  11. Therapeutic cloning: promises and issues

    PubMed Central

    Kfoury, Charlotte

    2007-01-01

    Advances in biotechnology necessitate both an understanding of scientific principles and ethical implications to be clinically applicable in medicine. In this regard, therapeutic cloning offers significant potential in regenerative medicine by circumventing immunorejection, and in the cure of genetic disorders when used in conjunction with gene therapy. Therapeutic cloning in the context of cell replacement therapy holds a huge potential for de novo organogenesis and the permanent treatment of Parkinson’s disease, Duchenne muscular dystrophy, and diabetes mellitus as shown by in vivo studies. Scientific roadblocks impeding advancement in therapeutic cloning are tumorigenicity, epigenetic reprogramming, mitochondrial heteroplasmy, interspecies pathogen transfer, low oocyte availability. Therapeutic cloning is also often tied to ethical considerations concerning the source, destruction and moral status of IVF embryos based on the argument of potential. Legislative and funding issues are also addressed. Future considerations would include a distinction between therapeutic and reproductive cloning in legislative formulations. PMID:18523539

  12. [Scientific ethics of human cloning].

    PubMed

    Valenzuela, Carlos Y

    2005-01-01

    True cloning is fission, budding or other types of asexual reproduction. In humans it occurs in monozygote twinning. This type of cloning is ethically and religiously good. Human cloning can be performed by twinning (TWClo) or nuclear transfer (NTClo). Both methods need a zygote or a nuclear transferred cell, obtained in vitro (IVTec). They are under the IVTec ethics. IVTecs use humans (zygotes, embryos) as drugs or things; increase the risk of malformations; increase development and size of abnormalities and may cause long-term changes. Cloning for preserving extinct (or almost extinct) animals or humans when sexual reproduction is not possible is ethically valid. The previous selection of a phenotype in human cloning violates some ethical principles. NTClo for reproductive or therapeutic purposes is dangerous since it increases the risk for nucleotide or chromosome mutations, de-programming or re-programming errors, aging or malignancy of the embryo cells thus obtained.

  13. FEN1 −69G>A and +4150G>T polymorphisms and breast cancer risk

    PubMed Central

    Rezaei, Maryam; Hashemi, Mohammad; Sanaei, Sara; Mashhadi, Mohammad Ali; Hashemi, Seyed Mehdi; Bahari, Gholamreza; Taheri, Mohsen

    2016-01-01

    Flap endonuclease 1 (FEN1), a DNA repair protein, is important in preventing carcinogenesis. Two functional germ line variants −69G>A (rs174538) and +4150G>T (rs4246215) in the FEN1 gene have been associated with risk of various types of cancer. The aim of the present study was to evaluate the possible impact of FEN1 polymorphisms on risk of breast cancer (BC) in a sample of Iranian subjects. The FEN1 −69G>A and +4150G>T polymorphisms were analyzed in a case-control study that included 266 BC patients and 225 healthy females. Polymerase chain reaction-restriction fragment length polymorphism analysis was used to genotype the variants. The findings demonstrated that the FEN1 −69G>A and +4150G>T polymorphisms were not associated with BC risk in co-dominant, dominant and recessive inheritance models. The findings indicated that GG/GT, GA/GG and GA/TT genotypes significantly decreased the risk of BC when compared with −69GG/+4150GG. Furthermore, haplotype analysis indicated that −69G/+4150T as well as −69A/+4150G significantly decreased the risk of BC compared with −69G/+4150G. Thus, these findings demonstrated that haplotypes of FEN1 −69G>A and +4150G>T polymorphisms decreased the risk of BC in an Iranian population. Further studies with larger sample sizes and different ethnicities are required to validate the present findings. PMID:27699013

  14. Multi-physiopathological consequences of the c.1392G>T CFTR mutation revealed by clinical and cellular investigations.

    PubMed

    Farhat, Raed; El-Seedy, Ayman; El-Moussaoui, Kamal; Pasquet, Marie-Claude; Adolphe, Catherine; Bieth, Eric; Languepin, Jeanne; Sermet-Gaudelus, Isabelle; Kitzis, Alain; Ladevèze, Véronique

    2015-02-01

    This study combines a clinical approach and multiple level cellular analyses to determine the physiopathological consequences of the c.1392G>T (p.Lys464Asn) CFTR exon 10 mutation, detected in a CF patient with a frameshift deletion in trans and a TG(11)T(5) in cis. Minigene experiment, with different TG(m)T(n) alleles, and nasal cell mRNA extracts were used to study the impact of c.1392G>T on splicing in both in cellulo and in vivo studies. The processing and localization of p.Lys464Asn protein were evaluated, in cellulo, by western blotting analyses and confocal microscopy. Clinical and channel exploration tests were performed on the patient to determine the exact CF phenotype profile and the CFTR chloride transport activity. c.1392G>T affects exon 10 splicing by inducing its complete deletion and encoding a frameshift transcript. The polymorphism TG(11)T(5) aggravates the effects of this mutation on aberrant splicing. Analysis of mRNA obtained from parental airway epithelial cells confirmed these in cellulo results. At the protein level the p.Lys464Asn protein showed neither maturated form nor membrane localization. Furthermore, the in vivo channel tests confirmed the absence of CFTR activity. Thus, the c.1392G>T mutation alone or in association with the TG repeats and the poly T tract revealed obvious impacts on splicing and CFTR protein processing and functionality. The c.[T(5); 1392G>T] complex allele contributes to the CF phenotype by affecting splicing and inducing a severe misprocessing defect. These results demonstrate that the classical CFTR mutations classification is not sufficient: in vivo and in cellulo studies of a possible complex allele in a patient are required to provide correct CFTR mutation classification, adequate medical counseling, and adapted therapeutic strategies.

  15. Outcomes of alpha 1,3-GT-knockout porcine heart transplants into a preclinical nonhuman primate model.

    PubMed

    Kim, H; Chee, H K; Yang, J; Hwang, S; Han, K H; Kang, J; Park, J H; Kim, J S; Lee, S J; Ock, S A; Park, M H; Park, K S; Lee, B C; Byeongchun, L; Cho, K; Noh, J; Park, W; Yun, I J; Ahn, C

    2013-10-01

    Solid organ xenotransplantation is a potential solution to current organ shortages in allotransplantation. We performed four heart transplantations from alpha1, 3-galactosyltransferase gene-knockout (GT-KO) pigs to cynomolgus monkeys and monitored immunological parameters before and after transplantation. After blood typing of the cynomolgus monkeys, we assessed the binding activity of immunoglobulin G (IgG) and IgM of monkey serum and serum toxicity toward porcine peripheral blood mononuclear cells (PBMCs) using flow cytometry. Immunosuppressive protocols consisted of anti-thymocyte globulin (25 mg/kg), rituximab (20 mg/kg), anti-CD154mAb (20 mg/kg), cobra venom factor (0.05 mg/kg), tacrolimus, and steroid. Cynomolgus monkeys with A or AB blood type with the lowest antibody binding and serum toxicity activity on porcine PBMCs were selected as recipients. Absolute numbers of CD3(+) T cells, CD20(+) B cells, and CD3(+)CD95(+) memory T cells in the peripheral blood were suppressed upto 24 days after transplantation. Interferon gamma production of T cells in response to porcine antigens were also significantly suppressed. Heart xenografts from GT-KO pigs survived for upto 24 days without pathologic evidence of rejection. We successfully performed 4 heart xenotransplantations using GT-KO pigs. We overcame hyperacute rejection by using GT-KO pigs, and all of the heart xenografts from the GT-KO pigs survived between 11 and 24 days without pathologic evidence of rejection, disseminated intravascular coagulation, or consumptive coagulopathy; however, we need to optimize protocols for immune modulation and postoperative care to attain long-term survival of solid organ xenografts. Copyright © 2013. Published by Elsevier Inc.

  16. The FSHB -211G>T variant attenuates serum FSH levels in the supraphysiological gonadotropin setting of Klinefelter syndrome.

    PubMed

    Busch, Alexander S; Tüttelmann, Frank; Zitzmann, Michael; Kliesch, Sabine; Gromoll, Jörg

    2015-05-01

    Klinefelter syndrome (47, XXY) is the most frequent genetic cause of male infertility and individuals share the endocrine hallmark of hypergonadotropic hypogonadism. Single-nucleotide polymorphisms located within the FSHB/FSHR gene were recently shown to impact serum follicle-stimulating hormone (FSH) levels and other reproductive parameters in men. The objective of this study was to analyse the effect of FSHB-211G>T (c.-280G>T, rs10835638) as well as FSHR c.2039G>A (rs6166) and FSHR c.-29G>A (rs1394205) on endocrine and reproductive parameters in untreated and testosterone-treated Klinefelter patients. Patients were retrospectively selected from the clientele attending a university-based andrology centre. A total of 309 non-mosaic Klinefelter individuals between 18 and 65 years were included and genotyped for the variants by TaqMan assays. The untreated group comprised 248 men, in which the FSHB -211G>T allele was significantly associated with the reduced serum follicle-stimulating hormone levels (-6.5 U/l per T allele, P=1.3 × 10(-3)). Testosterone treatment (n=150) abolished the observed association. When analysing patients before and under testosterone treatment (n=89), gonadotropin levels were similarly suppressed independently of the FSHB genotype. The FSHR polymorphisms did not exhibit any significant influence in any group, neither on the endocrine nor reproductive parameters. In conclusion, a hypergonadotropic setting such as Klinefelter syndrome does not mask the FSHB -211G>T genotype effects on the follicle-stimulating hormone serum levels. The impact was indeed more pronounced compared with normal or infertile men, whereas gonadotropin suppression under testosterone treatment seems to be independent of the genotype. Thus, the FSHB -211G>T genotype is a key determinant in the regulation of gonadotropins in different reproductive-endocrine pathopyhsiologies.

  17. Identification and in silico characterization of soybean trihelix-GT and bHLH transcription factors involved in stress responses

    PubMed Central

    Osorio, Marina Borges; Bücker-Neto, Lauro; Castilhos, Graciela; Turchetto-Zolet, Andreia Carina; Wiebke-Strohm, Beatriz; Bodanese-Zanettini, Maria Helena; Margis-Pinheiro, Márcia

    2012-01-01

    Environmental stresses caused by either abiotic or biotic factors greatly affect agriculture. As for soybean [Glycine max (L.) Merril], one of the most important crop species in the world, the situation is not different. In order to deal with these stresses, plants have evolved a variety of sophisticated molecular mechanisms, to which the transcriptional regulation of target-genes by transcription factors is crucial. Even though the involvement of several transcription factor families has been widely reported in stress response, there still is a lot to be uncovered, especially in soybean. Therefore, the objective of this study was to investigate the role of bHLH and trihelix-GT transcription factors in soybean responses to environmental stresses. Gene annotation, data mining for stress response, and phylogenetic analysis of members from both families are presented herein. At least 45 bHLH (from subgroup 25) and 63 trihelix-GT putative genes reside in the soybean genome. Among them, at least 14 bHLH and 11 trihelix-GT seem to be involved in responses to abiotic/biotic stresses. Phylogenetic analysis successfully clustered these with members from other plant species. Nevertheless, bHLH and trihelix-GT genes encompass almost three times more members in soybean than in Arabidopsis or rice, with many of these grouping into new clades with no apparent near orthologs in the other analyzed species. Our results represent an important step towards unraveling the functional roles of plant bHLH and trihelix-GT transcription factors in response to environmental cues. PMID:22802709

  18. Animal cloning: problems and prospects.

    PubMed

    Wells, D N

    2005-04-01

    An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred to the reproductive tracts of recipient cows result in healthy, longterm surviving clones. Of concern are the high losses throughout gestation, during birth and in the post-natal period through to adulthood. Many of the pregnancy losses relate to failure of the placenta to develop and function correctly. Placental dysfunction may also have an adverse influence on postnatal health. These anomalies are probably due to incorrect epigenetic reprogramming of the donor genome following NT, leading to inappropriate patterns of gene expression during the development of clones. Whilst some physiological tests on surviving clones suggest normality, other reports indicate a variety of post-natal clone-associated abnormalities. This variability in outcome may reflect species-specific and/or cloning methodological differences. Importantly, to date it appears that these clone-associated phenotypes are not transmitted to offspring following sexual reproduction. This indicates that they represent epigenetic errors, rather than genetic errors, which are corrected during gametogenesis. Whilst this needs confirmation at the molecular level, it provides initial confidence in the first application of NT in agriculture, namely, the production of small numbers of cloned sires from genetically elite bulls, for natural mating, to effectively disseminate genetic gain. In addition to the animal welfare concerns with the technology, the underlying health of the animals and the consequential effect on food safety are critical aspects that require investigation to gain regulatory and consumer

  19. Comparison of GT3X accelerometer and Yamax pedometer steps/day in a free-living sample of overweight and obese adults

    USDA-ARS?s Scientific Manuscript database

    The purpose of this study was to compare steps/day detected by the YAMAX SW-200 pedometer versus the Actigraph GT3X accelerometer in free-living adults. Daily YAMAX and GT3X steps were collected from a sample of 23 overweight and obese participants (78% female; age = 52.6 +/- 8.4 yr.; BMI = 31.0 +/-...

  20. Optimal cloning of pure states, testing single clones

    NASA Astrophysics Data System (ADS)

    Keyl, M.; Werner, R. F.

    1999-07-01

    We consider quantum devices for turning a finite number N of d-level quantum systems in the same unknown pure state σ into M>N systems of the same kind, in an approximation of the M-fold tensor product of the state σ. In a previous paper it was shown that this problem has a unique optimal solution, when the quality of the output is judged by arbitrary measurements, involving also the correlations between the clones. We show in this paper, that if the quality judgment is based solely on measurements of single output clones, there is again a unique optimal cloning device, which coincides with the one found previously.

  1. Human cloning: can it be made safe?

    PubMed

    Rhind, Susan M; Taylor, Jane E; De Sousa, Paul A; King, Tim J; McGarry, Michelle; Wilmut, Ian

    2003-11-01

    There are continued claims of attempts to clone humans using nuclear transfer, despite the serious problems that have been encountered in cloning other mammals. It is known that epigenetic and genetic mechanisms are involved in clone failure, but we still do not know exactly how. Human reproductive cloning is unethical, but the production of cells from cloned embryos could offer many potential benefits. So, can human cloning be made safe?

  2. Single-level optimization of a hybrid SOFC-GT power plant

    NASA Astrophysics Data System (ADS)

    Calise, F.; Dentice d'Accadia, M.; Vanoli, L.; von Spakovsky, M. R.

    The detailed synthesis/design optimization of a hybrid solid oxide fuel cell-gas turbine (SOFC-GT) power plant is presented in this paper. In the first part of the paper, the bulk-flow model used to simulate the plant is discussed. The performance of the centrifugal compressors and radial turbine is determined using maps, properly scaled in order to match the values required for mass flow rate and pressure ratio. Compact heat exchangers are simulated using Colburn and friction factor correlations. For the SOFC, the cell voltage versus current density curves (i.e. polarization curves) are generated on the basis of the Nernst potential and overvoltages. Validation of the SOFC polarization curves is accomplished with data available from Siemens Westinghouse. Both the steam-methane pre-reforming and internal reforming processes are modeled assuming the water-gas shift reaction to be equilibrium-controlled and the demethanization reactions to be kinetically controlled. Finally, a thermoeconomic model is developed by introducing capital cost functions for each plant component. The whole plant is first simulated for a fixed configuration. Then, a synthesis/design optimization of the plant is carried out using a traditional single-level approach. The results of the optimization are presented and discussed.

  3. Evolution of the Power Conversion Unit Design of the GT-MHR

    SciTech Connect

    Baxi, C.B.; Perez, E.; Shenoy, A.; Kostin, V.I.; Kodochigov, N.G.; Vasyaev, A.V.; Belov, S.E.; Golovko, V.F.

    2006-07-01

    General Atomics in the USA and Experimental Design Bureau of Machine Building (OKBM) in the Russian Federation are jointly developing a gas turbine modular helium reactor (GT-MHR). The 600 MW(t) reactor is cooled by helium at a pressure of 7 MPa. The power conversion unit (PCU) uses the reactor outlet temperature of 850 deg C in a direct Brayton cycle to achieve an efficiency of about 48%. The PCU consists of a gas turbine, a recuperator, a pre-cooler, a low-pressure compressor, an inter-cooler, and a high-pressure compressor. The turbo machine (TM), including the generator, is mounted on a single vertical shaft. The TM rotates at a speed of 4400 rpm. The asynchronous generator is connected to the turbine by a flexible coupling. The required grid frequency is achieved by a converter. All PCU components are enclosed in a single vessel. TM uses radial and axial electromagnetic bearings (EMB) for support. Catcher bearings (CB) are provided as redundant support for the TM rotor in case of EMBs failure. These design features were determined after a comprehensive study carried out over the last 10 years. This paper describes the evolution of the current PCU design and justification for the choices. (authors)

  4. A Novel Missense Mutation 224G>T (R75M) in SRY Coding Region Interferes with Nuclear Import and Results in 46, XY Complete Gonadal Dysgenesis

    PubMed Central

    He, Shanshan; Zhang, Tengfei; Yin, Chenxing; Chen, Yunping; Zheng, Shuqi; Zhang, Jixia; Li, Lin

    2016-01-01

    SRY-mutation-caused sex reversal is a rare disease and mostly associated with a de novo mutation since the patients with defective SRY is infertile. There are many reports about SRY-mutation associated 46, XY ovarian disorder of sex development (DSD), but few described their molecular mechanism. Here we report a de novo mutation 224G>T (R75M) in SRY associated with a phenotypic female, 46, XY karyotype and dysgerminoma. The wild and mutated SRY were cloned into recombinant plasmid and expressed in cells in vitro, the result showed the mutated SRY is greatly accumulated in cytoplasm while the wild type SRY is mostly localized in nucleus. To make sure no other genes were involved, we performed the trio-based whole exome sequencing using the DNA samples from the proband and the parents, and no mutations were identified especially in DHH, NR0B1, NR5A1, SOX9 and MAP3K1, indicating the de novo mutation in SRY is the single defect responsible for the female sex reversal. We also used bioinformatics simulation analysis to predict impact of the mutation on SRY function, and find the R75 in wild type SRY can form a hydrogen bond with serine at 91 (S91) that make the SRY protein well fit into the minor groove of target DNA, while the M75 in the mutated SRY can’t. Finally, we reviewed SRY mutations based on the available references and analyzed the mutation distribution patterns according to density and continuity, which may be useful for further study of the SRY structure, function, and its relatedness with DSD. PMID:28030592

  5. A Novel Missense Mutation 224G>T (R75M) in SRY Coding Region Interferes with Nuclear Import and Results in 46, XY Complete Gonadal Dysgenesis.

    PubMed

    Fan, Wufang; Wang, Bei; He, Shanshan; Zhang, Tengfei; Yin, Chenxing; Chen, Yunping; Zheng, Shuqi; Zhang, Jixia; Li, Lin

    2016-01-01

    SRY-mutation-caused sex reversal is a rare disease and mostly associated with a de novo mutation since the patients with defective SRY is infertile. There are many reports about SRY-mutation associated 46, XY ovarian disorder of sex development (DSD), but few described their molecular mechanism. Here we report a de novo mutation 224G>T (R75M) in SRY associated with a phenotypic female, 46, XY karyotype and dysgerminoma. The wild and mutated SRY were cloned into recombinant plasmid and expressed in cells in vitro, the result showed the mutated SRY is greatly accumulated in cytoplasm while the wild type SRY is mostly localized in nucleus. To make sure no other genes were involved, we performed the trio-based whole exome sequencing using the DNA samples from the proband and the parents, and no mutations were identified especially in DHH, NR0B1, NR5A1, SOX9 and MAP3K1, indicating the de novo mutation in SRY is the single defect responsible for the female sex reversal. We also used bioinformatics simulation analysis to predict impact of the mutation on SRY function, and find the R75 in wild type SRY can form a hydrogen bond with serine at 91 (S91) that make the SRY protein well fit into the minor groove of target DNA, while the M75 in the mutated SRY can't. Finally, we reviewed SRY mutations based on the available references and analyzed the mutation distribution patterns according to density and continuity, which may be useful for further study of the SRY structure, function, and its relatedness with DSD.

  6. Limitations on cloning in classical mechanics

    NASA Astrophysics Data System (ADS)

    Fenyes, Aaron

    2012-01-01

    In this paper, we show that a result precisely analogous to the traditional quantum no-cloning theorem holds in classical mechanics. This classical no-cloning theorem does not prohibit classical cloning, we argue, because it is based on a too-restrictive definition of cloning. Using a less popular, more inclusive definition of cloning, we give examples of classical cloning processes. We also prove that a cloning machine must be at least as complicated as the object it is supposed to clone.

  7. Wildlife conservation and reproductive cloning.

    PubMed

    Holt, William V; Pickard, Amanda R; Prather, Randall S

    2004-03-01

    Reproductive cloning, or the production of offspring by nuclear transfer, is often regarded as having potential for conserving endangered species of wildlife. Currently, however, low success rates for reproductive cloning limit the practical application of this technique to experimental use and proof of principle investigations. In this review, we consider how cloning may contribute to wildlife conservation strategies. The cloning of endangered mammals presents practical problems, many of which stem from the paucity of knowledge about their basic reproductive biology. However, situations may arise where resources could be targeted at recovering lost or under-represented genetic lines; these could then contribute to the future fitness of the population. Approaches of this type would be preferable to the indiscriminate generation of large numbers of identical individuals. Applying cloning technology to non-mammalian vertebrates may be more practical than attempting to use conventional reproductive technologies. As the scientific background to cloning technology was pioneered using amphibians, it may be possible to breed imminently threatened amphibians, or even restore extinct amphibian species, by the use of cloning. In this respect species with external embryonic development may have an advantage over mammals as developmental abnormalities associated with inappropriate embryonic reprogramming would not be relevant.

  8. Cloning operator and its applications

    NASA Astrophysics Data System (ADS)

    Voicu, Liviu I.; Myler, Harley R.; Toma, Cristian E.

    1998-03-01

    A novel genetic operator called cloning is introduced and tested in different applications of genetic algorithms. Essentially, the cloning monotonically increases the lengths of the chromosomes during the evolution. It is argued that, under these circumstances, the cloning operator can accommodate a multiresolution search strategy, where the search starts at coarser scales and is subsequently mapped to finer scales upon achieving some in-scale performance criteria. Although the practical implementation of cloning is application dependent, a few general requirements are stated. In the remainder of the paper, different implementations of the cloning operator are introduced and employed in distinct applications, namely, function optimization, object support reconstruction from the support of its autocorrelation and the shortest path problem in planar graphs. The first two cases present typical multiresolution approaches to search problems and their results show consistent improvements in convergence speed with respect to classical genetic algorithms. In the last problem, a cloning operator is incorporated in an evolutionary algorithm that builds a set of valid paths in a planar graph. It is demonstrated that cloning can enhance the ability of a genetic algorithm to explore the search space efficiently in some applications.

  9. Seamless Ligation Cloning Extract (SLiCE) cloning method.

    PubMed

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2014-01-01

    SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies.

  10. Molecular cloning of the avian erythroblastosis virus genome and recovery of oncogenic virus by transfection of chicken cells.

    PubMed Central

    Vennström, B; Fanshier, L; Moscovici, C; Bishop, J M

    1980-01-01

    Avian erythroblastosis virus (AEV) causes erythroblastosis and sarcomas in birds and transforms both erythroblasts and fibroblasts to neoplastic phenotypes in culture. The viral genetic locus required for oncogenesis by AEV is at present poorly defined; moreover, we know very little of the mechanism of tumorigenesis by the virus. To facilitate further analysis of these problems, we used molecular cloning to isolate the genome of AEV as recombinant DNA in a procaryotic vector. The identity of the isolated DNA was verified by mapping with restriction endonucleases and by tests for biological activity. The circular form of unintegrated AEV DNA was purified from synchronously infected quail cells and cloned into the EcoRI site of lambda gtWES x B. A restriction endonuclease cleavage map was established. By hybridization with complementary DNA probes representing specific parts of avian retrovirus genomes, the restriction map of the cloned AEV DNAs was correlated with a genetic map. These data show that nucleotide sequences unique to AEV comprise at least 50% of the genome and are located approximately in the middle of the AEV genome. Our data confirm and extend previous descriptions of the AEV genome obtained by other procedures. We studied in detail two recombinant clones containing AEV DNA: the topography of the viral DNA in the two clones was virtually identical, except that one clone apparently contained two copies of the terminal redundancy that occurs in linear viral DNA isolated from infected cells; the other clone probably contained only one copy of the redundant sequence. To recover infectious virus from the cloned DNA, we developed a procedure for transfection that compensated for the defectiveness of AEV in replication. We accomplished this by ligating cloned AEV DNA to the cloned DNA of a retrovirus (Rous-associated virus type 1) whose genome could complement the deficiencies of AEV. Ligation of the two viral DNAs was facilitated by using a neutral fragment

  11. Biomimetic Cloning of Quantum Observables

    NASA Astrophysics Data System (ADS)

    Alvarez-Rodriguez, U.; Sanz, M.; Lamata, L.; Solano, E.

    2014-05-01

    We propose a bio-inspired sequential quantum protocol for the cloning and preservation of the statistics associated to quantum observables of a given system. It combines the cloning of a set of commuting observables, permitted by the no-cloning and no-broadcasting theorems, with a controllable propagation of the initial state coherences to the subsequent generations. The protocol mimics the scenario in which an individual in an unknown quantum state copies and propagates its quantum information into an environment of blank qubits. Finally, we propose a realistic experimental implementation of this protocol in trapped ions.

  12. Therapeutic cloning in the mouse

    PubMed Central

    Mombaerts, Peter

    2003-01-01

    Nuclear transfer technology can be applied to produce autologous differentiated cells for therapeutic purposes, a concept termed therapeutic cloning. Countless articles have been published on the ethics and politics of human therapeutic cloning, reflecting the high expectations from this new opportunity for rejuvenation of the aging or diseased body. Yet the research literature on therapeutic cloning, strictly speaking, is comprised of only four articles, all in the mouse. The efficiency of derivation of embryonic stem cell lines via nuclear transfer is remarkably consistent among these reports. However, the efficiency is so low that, in its present form, the concept is unlikely to become widespread in clinical practice. PMID:12949262

  13. Biomimetic Cloning of Quantum Observables

    PubMed Central

    Alvarez-Rodriguez, U.; Sanz, M.; Lamata, L.; Solano, E.

    2014-01-01

    We propose a bio-inspired sequential quantum protocol for the cloning and preservation of the statistics associated to quantum observables of a given system. It combines the cloning of a set of commuting observables, permitted by the no-cloning and no-broadcasting theorems, with a controllable propagation of the initial state coherences to the subsequent generations. The protocol mimics the scenario in which an individual in an unknown quantum state copies and propagates its quantum information into an environment of blank qubits. Finally, we propose a realistic experimental implementation of this protocol in trapped ions. PMID:24809937

  14. Molecular Cloning of Adenosinediphosphoribosyl Transferase.

    DTIC Science & Technology

    1987-09-08

    AD-RIB5 458 NOLECULNA CLONING OF AOENOSINEDXPNOSPHORIBOSyL 1/1 TRNSFERASEMU CAILIFORNIA UNIV SRN FRANCISCO E KUN US SEP 8? WFOSR-TR-87-0982 SWFOSR-B5...ACCESSION NO.D,. 03261102F 2312 A~5 11. TITLE (include Securqt Classification) 0 Molecular Cloning of Adenosinediphosphoribosyl Transferase 12. PERSONAL...I’:- AFOSR.Tlt. 8 7 - 0 9 8,2 0IL * pi AFOSR- 85 -0377 PROGRESS REPORT Molecular Cloning of Adenosinediphosphoribosyl Transferase 5." Period of

  15. Human therapeutic cloning (NTSC): applying research from mammalian reproductive cloning.

    PubMed

    French, Andrew J; Wood, Samuel H; Trounson, Alan O

    2006-01-01

    Human therapeutic cloning or nuclear transfer stem cells (NTSC) to produce patient-specific stem cells, holds considerable promise in the field of regenerative medicine. The recent withdrawal of the only scientific publications claiming the successful generation of NTSC lines afford an opportunity to review the available research in mammalian reproductive somatic cell nuclear transfer (SCNT) with the goal of progressing human NTSC. The process of SCNT is prone to epigenetic abnormalities that contribute to very low success rates. Although there are high mortality rates in some species of cloned animals, most surviving clones have been shown to have normal phenotypic and physiological characteristics and to produce healthy offspring. This technology has been applied to an increasing number of mammals for utility in research, agriculture, conservation, and biomedicine. In contrast, attempts at SCNT to produce human embryonic stem cells (hESCs) have been disappointing. Only one group has published reliable evidence of success in deriving a cloned human blastocyst, using an undifferentiated hESC donor cell, and it failed to develop into a hESC line. When optimal conditions are present, it appears that in vitro development of cloned and parthenogenetic embryos, both of which may be utilized to produce hESCs, may be similar to in vitro fertilized embryos. The derivation of ESC lines from cloned embryos is substantially more efficient than the production of viable offspring. This review summarizes developments in mammalian reproductive cloning, cell-to-cell fusion alternatives, and strategies for oocyte procurement that may provide important clues facilitating progress in human therapeutic cloning leading to the successful application of cell-based therapies utilizing autologous hESC lines.

  16. Methylotroph cloning vehicle

    DOEpatents

    Hanson, Richard S.; Allen, Larry N.

    1989-04-25

    A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C.sub.1 -utilizing host and in a C.sub.1 -utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C.sub.1 -utilizing host to the C.sub.1 -utilizing host; DNA providing resistance to two antibiotics to which the wild-type C.sub.1 -utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C.sub.1 -utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C.sub.1 -utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C.sub.1 -utilizing (e.g., E. coli) host, and then conjugated with a selected C.sub.1 -utilizing mutant. Resistance to the other antibiotic by the mutant is a marker of the conjugation. Other phenotypical changes in the mutant, e.g., loss of an auxotrophic trait, is attributed to the C.sub.1 gene. The vector is also used to inactivate genes whose protein products catalyze side reactions that divert compounds from a biosynthetic pathway to a desired product, thereby producing an organism that makes the desired product in higher yields.

  17. Factors influencing uptake of measles, mumps and rubella (MMR) immunization in site-dwelling Gypsy, Roma and Traveller (G&T) communities: a qualitative study of G&T parents' beliefs and experiences.

    PubMed

    Newton, P; Smith, D M

    2017-07-01

    Increasing immunization in the Gypsy, Roma and Traveller (G&T) community is a key priority for improving health outcomes in this community. This study aimed to explore G&T parents: (1) beliefs about childhood immunization; (2) beliefs about the risks of immunization and non-immunization; (3) perceived obstacles to, and facilitators of, immunization and (4) views on increasing immunization levels. A cross-sectional, qualitative study was conducted comprising of five focus groups with 16 site-dwelling G&T women with pre-school aged children. Data were transcribed verbatim and analysed thematically. Five main themes were identified: Lay understandings of causation and risk; Timing of immunization; Children being perceived as vulnerable; The fit between lifestyle and healthcare provision; The impact of living with a high burden of disease. Understanding of the risks and benefits of measles, mumps and rubella immunization did not differ significantly from the wider population or those promoted by the health service. The majority of barriers stemmed from living with the effects of long-standing health inequalities, which posed further barriers to accessing immunization, and shaped beliefs about immunization. Factors facilitating uptake occurred where access to immunization services was made flexible, e.g. immunization on traveller sites. © 2017 John Wiley & Sons Ltd.

  18. Human Cloning: Let's Discuss It.

    ERIC Educational Resources Information Center

    Taras, Loretta; Stavroulakis, Anthea M.; Ortiz, Mary T.

    1999-01-01

    Describes experiences with holding discussions on cloning at a variety of levels in undergraduate biology courses. Discusses teaching methods used and student reactions to the discussions. Contains 12 references. (WRM)

  19. A Clone of Your Own.

    ERIC Educational Resources Information Center

    Bilodeau, Kirsten

    1997-01-01

    Describes an activity used at the Washington Park Arboretum that helps students understand cloning through plant propagation. Students also learn how to make a pot from recycled newspapers and how to make soil that is appropriate for the plants. (DDR)

  20. Cloning of a quantum measurement

    SciTech Connect

    Bisio, Alessandro; D'Ariano, Giacomo Mauro; Perinotti, Paolo; Sedlak, Michal

    2011-10-15

    We analyze quantum algorithms for cloning of a quantum measurement. Our aim is to mimic two uses of a device performing an unknown von Neumann measurement with a single use of the device. When the unknown device has to be used before the bipartite state to be measured is available we talk about 1{yields}2 learning of the measurement, otherwise the task is called 1{yields}2 cloning of a measurement. We perform the optimization for both learning and cloning for arbitrary dimension d of the Hilbert space. For 1{yields}2 cloning we also propose a simple quantum network that achieves the optimal fidelity. The optimal fidelity for 1{yields}2 learning just slightly outperforms the estimate and prepare strategy in which one first estimates the unknown measurement and depending on the result suitably prepares the duplicate.

  1. Are cloned quantum states macroscopic?

    PubMed

    Fröwis, F; Dür, W

    2012-10-26

    We study quantum states produced by optimal phase covariant quantum cloners. We argue that cloned quantum superpositions are not macroscopic superpositions in the spirit of Schrödinger's cat, despite their large particle number. This is indicated by calculating several measures for macroscopic superpositions from the literature, as well as by investigating the distinguishability of the two superposed cloned states. The latter rapidly diminishes when considering imperfect detectors or noisy states and does not increase with the system size. In contrast, we find that cloned quantum states themselves are macroscopic, in the sense of both proposed measures and their usefulness in quantum metrology with an optimal scaling in system size. We investigate the applicability of cloned states for parameter estimation in the presence of different kinds of noise.

  2. A Clone of Your Own.

    ERIC Educational Resources Information Center

    Bilodeau, Kirsten

    1997-01-01

    Describes an activity used at the Washington Park Arboretum that helps students understand cloning through plant propagation. Students also learn how to make a pot from recycled newspapers and how to make soil that is appropriate for the plants. (DDR)

  3. Human cloning and 'posthuman' society.

    PubMed

    Blackford, Russell

    2005-01-01

    Since early 1997, when the creation of Dolly the sheep by somatic cell nuclear transfer was announced in Nature, numerous government reports, essays, articles and books have considered the ethical problems and policy issues surrounding human reproductive cloning. In this article, I consider what response a modern liberal society should give to the prospect of human cloning, if it became safe and practical. Some opponents of human cloning have argued that permitting it would place us on a slippery slope to a repugnant future society, comparable to that portrayed in Aldous Huxley's novel, Brave New World. I conclude that, leaving aside concerns about safety, none of the psychological or social considerations discussed in this article provides an adequate policy justification for invoking the state's coercive powers to prevent human cloning.

  4. Animal Cloning and Food Safety

    MedlinePlus

    ... Products For Consumers Home For Consumers Consumer Updates Animal Cloning and Food Safety Share Tweet Linkedin Pin ... safe to eat as food from conventionally bred animals. This conclusion stems from an extensive study of ...

  5. Human Cloning: Let's Discuss It.

    ERIC Educational Resources Information Center

    Taras, Loretta; Stavroulakis, Anthea M.; Ortiz, Mary T.

    1999-01-01

    Describes experiences with holding discussions on cloning at a variety of levels in undergraduate biology courses. Discusses teaching methods used and student reactions to the discussions. Contains 12 references. (WRM)

  6. Unique motifs identify PIG-A proteins from glycosyltransferases of the GT4 family

    PubMed Central

    2008-01-01

    Background The first step of GPI anchor biosynthesis is catalyzed by PIG-A, an enzyme that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to phosphatidylinositol. This protein is present in all eukaryotic organisms ranging from protozoa to higher mammals, as part of a larger complex of five to six 'accessory' proteins whose individual roles in the glycosyltransferase reaction are as yet unclear. The PIG-A gene has been shown to be an essential gene in various eukaryotes. In humans, mutations in the protein have been associated with paroxysomal noctural hemoglobuinuria. The corresponding PIG-A gene has also been recently identified in the genome of many archaeabacteria although genes of the accessory proteins have not been discovered in them. The present study explores the evolution of PIG-A and the phylogenetic relationship between this protein and other glycosyltransferases. Results In this paper we show that out of the twelve conserved motifs identified by us eleven are exclusively present in PIG-A and, therefore, can be used as markers to identify PIG-A from newly sequenced genomes. Three of these motifs are absent in the primitive eukaryote, G. lamblia. Sequence analyses show that seven of these conserved motifs are present in prokaryote and archaeal counterparts in rudimentary forms and can be used to differentiate PIG-A proteins from glycosyltransferases. Using partial least square regression analysis and data involving presence or absence of motifs in a range of PIG-A and glycosyltransferases we show that (i) PIG-A may have evolved from prokaryotic glycosyltransferases and lipopolysaccharide synthases, members of the GT4 family of glycosyltransferases and (ii) it is possible to uniquely classify PIG-A proteins versus glycosyltransferases. Conclusion Besides identifying unique motifs and showing that PIG-A protein from G. lamblia and some putative PIG-A proteins from archaebacteria are evolutionarily closer to glycosyltransferases, these studies

  7. Concurrent validity of the MOX activity monitor compared to the ActiGraph GT3X.

    PubMed

    van der Weegen, Sanne; Essers, Hans; Spreeuwenberg, Marieke; Verwey, Renée; Tange, Huibert; de Witte, Luc; Meijer, Kenneth

    2015-04-01

    The It's LiFe! monitoring and feedback tool embedded in primary care practice is promising in helping people to achieve an active lifestyle. This new tool consists of an activity monitor (the MOX), which is connected to a smartphone application and to a Web service for patients and care providers. The aim of this study was to develop thresholds for the moderate and vigorous activity categories and examine the concurrent validity of the MOX in relation to the ActiGraph (Pensacola, FL) GT3X in healthy participants and chronically ill patients (chronic obstructive pulmonary disease and type 2 diabetes) in a laboratory situation and during daily living. Participants wore the two activity monitors simultaneously on the lower back. An incremental treadmill protocol was executed by 8 healthy adults and 10 patients. For daily living measurements, 15 healthy adults and 12 patients wore the devices for 6-7 days. Daily living data were corrected for non-wear time, using diary information. On the treadmill there was an excellent correlation between the ActiGraph and MOX counts (mean r=0.99 in healthy participants and mean r=0.98 in patients). Correlation during daily living was moderate (mean r=0.72) in healthy adults and good (mean r=0.82) in patients. Bland-Altman plots showed no perfect agreement between the two devices in minutes per category. However, a histogram of misclassified minutes showed that misclassification occurred around category thresholds. The MOX is capable of measuring physical activity and can be used in the It's LiFe!

  8. Calibration of ActiGraph GT3X, Actical and RT3 accelerometers in adolescents.

    PubMed

    Romanzini, Marcelo; Petroski, Edio Luiz; Ohara, David; Dourado, Antonio Carlos; Reichert, Felipe Fossat

    2014-01-01

    The objective of this study was to develop count cut-points for three different accelerometer models: ActiGraph GT3X, RT3 and Actical to accurately classify physical activity intensity levels in adolescents. Seventy-nine adolescents (10-15 years) participated in this study. Accelerometers and oxygen consumption ([Formula: see text]) data were collected at rest and during 11 physical activities of different intensities. Accelerometers were worn on the waist and [Formula: see text] was measured by a portable metabolic system: Cosmed K4b2. Receiver operating characteristic (ROC) curves were used to determine cut-points. Cut-points for sedentary (SED), moderate-to-vigorous (MVPA) and vigorous-intensity physical activity (VPA) were 46, 607 and 818 counts·15s(-1) to the vertical axis of ActiGraph; 180, 757 and 1112 counts·15s(-1) to the vector magnitude of ActiGraph; 17, 441 and 873 counts·15s(-1) for Actical; and 5.6, 20.4 and 32.2 counts·s(-1) for RT3, respectively. For all three accelerometer models, there was an almost perfect discrimination of SED and MVPA (ROC >0.97) and an excellent discrimination of VPA (ROC>0.90) observed. Areas under the ROC curves indicated better discrimination of MVPA by ActiGraph (AUC=0.994) and Actical (AUC=0.993) when compared to RT3 (AUC=0.983). The cut-points developed in this study for the ActiGraph (vector magnitude), RT3 and Actical accelerometer models can be used to monitor physical activity level of adolescents.

  9. [Effects of nitriles and amides on the growth and the nitrile hydratase activity of the Rhodococcus sp. strain gt1].

    PubMed

    Maksimov, A Iu; Kuznetsova, M V; Ovechkina, G V; Kozlov, S V; Maksimova, Iu G; Demakov, V A

    2003-01-01

    Effects of some nitriles and amides, as well as glucose and ammonium, on the growth and the nitrile hydratase (EC 4.2.1.84) activity of the Rhodococcus sp. strain gt1 isolated from soil were studied. The activity of nitrile hydratase mainly depended on carbon and nitrogen supply to cells. The activity of nitrile hydratase was high in the presence of glucose and ammonium at medium concentrations and decreased at concentrations of glucose more than 0.3%. Saturated unsubstituted aliphatic nitriles and amides were found to be a good source of nitrogen and carbon. However, the presence of nitriles and amides in the medium was not absolutely necessary for the expression of the activity of nitrile hydratase isolated from the Rhodococcus sp. strain gt1.

  10. GT-MHR COMMERCIALIZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD AUGUST 1 THROUGH AUGUST 31, 2003

    SciTech Connect

    SHENOY, A.S.

    2003-08-01

    OAK A271 GT-MHR COMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD AUGUST 1 THROUGH AUGUST 31, 2003. In the process of fabricating the MHR-1 irradiation test capsule, Petten has advised that three thermocouples (out of 24) and the Self Powered Neutron detector were damaged during high temperature brazing with the upper capsule lid. Procurement of new TCs and SPN is in process but there will be a delay in the irradiation test of about nine weeks. Startup of the irradiation is now projected to be July or August 2004. In preparation for performing the nuclear design analysis activities required by the advanced fuel studies task, a complete 3-D nuclear design analysis is first being performed of the GT-MHR reference design. This will serve as the baseline for studies of the advanced fuel nuclear design performance.

  11. Methylotroph cloning vehicle

    DOEpatents

    Hanson, R.S.; Allen, L.N.

    1989-04-25

    A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C[sub 1]-utilizing host and in a C[sub 1]-utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C[sub 1]-utilizing host to the C[sub 1]-utilizing host; DNA providing resistance to two antibiotics to which the wild-type C[sub 1]-utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C[sub 1]-utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C[sub 1]-utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C[sub 1]-utilizing (e.g., E. coli) host, and then conjugated with a selected C[sub 1]-utilizing mutant. Resistance to the other antibiotic by the mutant is a marker of the conjugation. Other phenotypical changes in the mutant, e.g., loss of an auxotrophic trait, is attributed to the C[sub 1] gene. The vector is also used to inactivate genes whose protein products catalyze side reactions that divert compounds from a biosynthetic pathway to a desired product, thereby producing an organism that makes the desired product in higher yields. 3 figs.

  12. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research.

  13. Cloning goes to the movies.

    PubMed

    Cormick, Craig

    2006-10-01

    Public attitude research conducted by Biotechnology Australia shows that one of the major sources of information on human reproductive cloning is movies. Traditionally, understanding of new and emerging technologies has come through the mass media but human cloning, being so widely addressed through the popular culture of movies, is more effectively defined by Hollywood than the news media or science media. But how well are the science and social issues of cloning portrayed in box office hits such as The Island, Multiplicity, Star Wars: Attack of the Clones and Jurassic Park? These movies have enormous reach and undoubted influence, and are therefore worth analyzing in some detail. This study looks at 33 movies made between 1971 and 2005 that address human reproductive cloning, and it categorizes the films based on their genre and potential influence. Yet rather than simply rating the quality of the science portrayed, the study compares the key messages in these movies with public attitudes towards cloning, to examine the correlations.

  14. Artificial cloning of domestic animals

    PubMed Central

    Keefer, Carol L.

    2015-01-01

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research. PMID:26195770

  15. Islamic perspectives on human cloning.

    PubMed

    Sadeghi, Mahmoud

    2007-01-01

    The present paper seeks to assess various views from Islamic jurists relating to human cloning, which is one of the controversial topics in the recent past. Taking Islamic jurisprudence principles, such as the rule of necessity for self preservation and respect for human beings, the rule of la darar wa la dirar ('the necessity to refrain from causing harm to oneself and others') and the rule of usr wa haraj, one may indicate that if human cloning could not be prohibited, as such, it could still be opposed because it gives way to various harmful consequences, which include family disorder, chaos in the clone's family relationships, physical and mental diseases for clones and suffering of egg donors and surrogate mothers. However with due attention to the fact that the reasons behind the prohibition of abortion only restrict the destruction of human embryos in their post-implantation stages, human cloning for biomedical research and exploitation of stem cells from cloned embryos at the blastocyst stage for therapeutic purposes would be acceptable.

  16. Structural evidence of a passive base-flipping mechanism for AGT, an unusual GT-B glycosyltransferase.

    PubMed

    Larivière, Laurent; Sommer, Nicole; Moréra, Solange

    2005-09-09

    The Escherichia coli T4 bacteriophage uses two glycosyltransferases to glucosylate and thus protect its DNA: the retaining alpha-glucosyltransferase (AGT) and the inverting beta-glucosyltransferase (BGT). They glucosylate 5-hydroxymethyl cytosine (5-HMC) bases of duplex DNA using UDP-glucose as the sugar donor to form an alpha-glucosidic linkage and a beta-glucosidic linkage, respectively. Five structures of AGT have been determined: a binary complex with the UDP product and four ternary complexes with UDP or UDP-glucose and oligonucleotides containing an A:G, HMU:G (hydroxymethyl uracyl) or AP:G (apurinic/apyrimidinic) mismatch at the target base-pair. AGT adopts the GT-B fold, one of the two folds known for GTs. However, while the sugar donor binding mode is classical for a GT-B enzyme, the sugar acceptor binding mode is unexpected and breaks the established consensus: AGT is the first GT-B enzyme that predominantly binds both the sugar donor and acceptor to the C-terminal domain. Its active site pocket is highly similar to four retaining GT-B glycosyltransferases (trehalose-6-phosphate synthase, glycogen synthase, glycogen and maltodextrin phosphorylases) strongly suggesting a common evolutionary origin and catalytic mechanism for these enzymes. Structure-guided mutagenesis and kinetic analysis do not permit identification of a nucleophile residue responsible for a glycosyl-enzyme intermediate for the classical double displacement mechanism. Interestingly, the DNA structures reveal partially flipped-out bases. They provide evidence for a passive role of AGT in the base-flipping mechanism and for its specific recognition of the acceptor base.

  17. Scrapie protein degradation by cysteine proteases in CD11c+ dendritic cells and GT1-1 neuronal cells.

    PubMed

    Luhr, Katarina M; Nordström, Elin K; Löw, Peter; Ljunggren, Hans-Gustaf; Taraboulos, Albert; Kristensson, Krister

    2004-05-01

    Dendritic cells (DC) of the CD11c(+) myeloid phenotype have been implicated in the spread of scrapie in the host. Previously, we have shown that CD11c(+) DC can cause a rapid degradation of proteinase K-resistant prion proteins (PrP(Sc)) in vitro, indicating a possible role of these cells in the clearance of PrP(Sc). To determine the mechanisms of PrP(Sc) degradation, CD11c(+) DC that had been exposed to PrP(Sc) derived from a neuronal cell line (GT1-1) infected with scrapie (ScGT1-1) were treated with a battery of protease inhibitors. Following treatment with the cysteine protease inhibitors (2S,3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane (E-64c), its ethyl ester (E-64d), and leupeptin, the degradation of PrP(Sc) was inhibited, while inhibitors of serine and aspartic and metalloproteases (aprotinin, pepstatin, and phosphoramidon) had no effect. An endogenous degradation of PrP(Sc) in ScGT1-1 cells was revealed by inhibiting the expression of cellular PrP (PrP(C)) by RNA interference, and this degradation could also be inhibited by the cysteine protease inhibitors. Our data show that PrP(Sc) is proteolytically cleaved preferentially by cysteine proteases in both CD11c(+) DC and ScGT1-1 cells and that the degradation of PrP(Sc) by proteases is different from that of PrP(C). Interference by protease inhibitors with DC-induced processing of PrP(Sc) has the potential to modify prion spread, clearance, and immunization in a host.

  18. Draft Genome Sequence of Kluyvera intestini Strain GT-16 Isolated from the Stomach of a Patient with Gastric Cancer

    PubMed Central

    Tetz, Victor

    2016-01-01

    Here, we report the complete genome sequence of the novel, non-spore-forming Kluyvera intestini strain GT-16, isolated from the stomach of a patient with gastric cancer. The genome is 5,868,299 bp in length with a G+C content of 53.0%. It possesses 5,350 predicted protein-coding genes encoding virulence factors and antibiotic resistance proteins. PMID:28007864

  19. DOS cones along atomic chains

    NASA Astrophysics Data System (ADS)

    Kwapiński, Tomasz

    2017-03-01

    The electron transport properties of a linear atomic chain are studied theoretically within the tight-binding Hamiltonian and the Green’s function method. Variations of the local density of states (DOS) along the chain are investigated. They are crucial in scanning tunnelling experiments and give important insight into the electron transport mechanism and charge distribution inside chains. It is found that depending on the chain parity the local DOS at the Fermi level can form cone-like structures (DOS cones) along the chain. The general condition for the local DOS oscillations is obtained and the linear behaviour of the local density function is confirmed analytically. DOS cones are characterized by a linear decay towards the chain which is in contrast to the propagation properties of charge density waves, end states and Friedel oscillations in one-dimensional systems. We find that DOS cones can appear due to non-resonant electron transport, the spin–orbit scattering or for chains fabricated on a substrate with localized electrons. It is also shown that for imperfect chains (e.g. with a reduced coupling strength between two neighboring sites) a diamond-like structure of the local DOS along the chain appears.

  20. Thermo-economic comparative analysis of gas turbine GT10 integrated with air and steam bottoming cycle

    NASA Astrophysics Data System (ADS)

    Czaja, Daniel; Chmielnak, Tadeusz; Lepszy, Sebastian

    2014-12-01

    A thermodynamic and economic analysis of a GT10 gas turbine integrated with the air bottoming cycle is presented. The results are compared to commercially available combined cycle power plants based on the same gas turbine. The systems under analysis have a better chance of competing with steam bottoming cycle configurations in a small range of the power output capacity. The aim of the calculations is to determine the final cost of electricity generated by the gas turbine air bottoming cycle based on a 25 MW GT10 gas turbine with the exhaust gas mass flow rate of about 80 kg/s. The article shows the results of thermodynamic optimization of the selection of the technological structure of gas turbine air bottoming cycle and of a comparative economic analysis. Quantities are determined that have a decisive impact on the considered units profitability and competitiveness compared to the popular technology based on the steam bottoming cycle. The ultimate quantity that can be compared in the calculations is the cost of 1 MWh of electricity. It should be noted that the systems analyzed herein are power plants where electricity is the only generated product. The performed calculations do not take account of any other (potential) revenues from the sale of energy origin certificates. Keywords: Gas turbine air bottoming cycle, Air bottoming cycle, Gas turbine, GT10

  1. Reduction of Hexavalent Chromium by Green Tea Polyphenols and Green Tea Nano Zero-Valent Iron (GT-nZVI).

    PubMed

    Chrysochoou, M; Reeves, K

    2017-03-01

    This study reports on the direct reduction of hexavalent chromium [Cr(VI)] by green tea polyphenols, including a green tea solution and pure epigallocatechin gallate (EGCG) solution. A linear trend was observed between the amount of reduced Cr(VI) and the amount of added polyphenols. The green tea solution showed a continued decrease in the observed stoichiometry with increasing pH, from a maximum of 1.4 mol per gallic acid equivalent (GAE) of green tea at pH 2.5, to 0.2 mol/GAE at pH 8.8. The EGCG solution exhibited different behavior, with a maximum stoichiometry of 2 at pH 7 and minimum of 1.6 at pH 4.4 and 8.9. When green tea was used to first react with Fe(3+) and form GT-nZVI, the amount of Cr(VI) reduced by a certain volume of GT-nZVI was double compared to green tea, and 6 times as high considering that GT-nZVI only contains 33 % green tea.

  2. Experimental study of cesium 5D+5D-&gt;6S+(nL=9D,11S,7F) energy pooling collisions

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Dai, Kang; Shen, Yifan

    2007-07-01

    We report experimentally the measured rate coefficients for the energy pooling (EP) collisions process Cs(5D)+Cs(5D)-&gt;Cs(6S)+Cs(nL=9D,11S,7F) in cesium densities of 10^(16)-10^(17) cm^(-3). The 5D state was populated via 8S-&gt;7P-&gt;5D spontaneous emission following two-step pumping 6S-&gt;6P_(3/2)-&gt;8S. Since the 5D-&gt;6P (3.0-3.6 microns) fluorescence could not be detected in this experiment, we carried out a relative measurement for the process 6P+5D-&gt;6S+7D. The excited-atom density and spatial distribution were mapped by monitoring the absorption of a counterpropagating single-mode laser beam, tuned to 6P_(3/2)-&gt;9S_(1/2) transition, which could be translated parallelly to the pump beam. The excited atom densities have been combined with the measured fluorescence ratios to yield EP rate coefficients. The average values for nL=9D,11S and 7F are 8.0+-4.0, 7.0+-3.5, and 9.3+-4.6 (in units of 10^(-10) cm3/s), respectively. Influence of the energy transfer process 11S+6S-&gt;7F+6S on the rate coefficients k_(11S) and k_(7F) is also discussed.

  3. [Cloning of vertebrates: successes and problems].

    PubMed

    Koniukhov, B V

    1997-12-01

    Cloning of vertebrates, in particular, amphibians and mammals, is discussed. In the last decade, significant progress was made cloning mammals, while cloning of adult amphibians remained problematical. Low-traumatic methods of enucleation of recipient oocytes and transplantation of donor nuclei were worked out. In 1997, an adult sheep was cloned in Great Britain, thus demonstrating the possibility of cloning adult mammals. However, methods of cloning mammals need improvement because of the high lethality of reconstructed embryos (nuclear transplants). The use of in vitro cultured low-differentiated stem cells to obtain donor nuclei seems promising. Works on human cloning are not expedient in the near future because of technical and ethical aspects.

  4. GT-MSOCC - A domain for research on human-computer interaction and decision aiding in supervisory control systems. [Georgia Tech - Multisatellite Operations Control Center

    NASA Technical Reports Server (NTRS)

    Mitchell, Christine M.

    1987-01-01

    The Georgia Tech-Multisatellite Operations Control Center (GT-MSOCC), a real-time interactive simulation of the operator interface to a NASA ground control system for unmanned earth-orbiting satellites, is described. The GT-MSOCC program for investigating a range of modeling, decision aiding, and workstation design issues related to the human-computer interaction is discussed. A GT-MSOCC operator function model is described in which operator actions, both cognitive and manual, are represented as the lowest level discrete control network nodes, and operator action nodes are linked to information needs or system reconfiguration commands.

  5. GT-MSOCC - A domain for research on human-computer interaction and decision aiding in supervisory control systems. [Georgia Tech - Multisatellite Operations Control Center

    NASA Technical Reports Server (NTRS)

    Mitchell, Christine M.

    1987-01-01

    The Georgia Tech-Multisatellite Operations Control Center (GT-MSOCC), a real-time interactive simulation of the operator interface to a NASA ground control system for unmanned earth-orbiting satellites, is described. The GT-MSOCC program for investigating a range of modeling, decision aiding, and workstation design issues related to the human-computer interaction is discussed. A GT-MSOCC operator function model is described in which operator actions, both cognitive and manual, are represented as the lowest level discrete control network nodes, and operator action nodes are linked to information needs or system reconfiguration commands.

  6. Local cloning of two product states

    SciTech Connect

    Ji Zhengfeng; Feng Yuan; Ying Mingsheng

    2005-09-15

    Local quantum operations and classical communication (LOCC) put considerable constraints on many quantum information processing tasks such as cloning and discrimination. Surprisingly, however, discrimination of any two pure states survives such constraints in some sense. We show that cloning is not that lucky; namely, probabilistic LOCC cloning of two product states is strictly less efficient than global cloning. We prove our result by giving explicitly the efficiency formula of local cloning of any two product states.

  7. Local cloning of entangled states

    NASA Astrophysics Data System (ADS)

    Gheorghiu, Vlad; Yu, Li; Cohen, Scott M.

    2010-08-01

    We investigate the conditions under which a set S of pure bipartite quantum states on a D×D system can be locally cloned deterministically by separable operations, when at least one of the states is full Schmidt rank. We allow for the possibility of cloning using a resource state that is less than maximally entangled. Our results include that: (i) all states in S must be full Schmidt rank and equally entangled under the G-concurrence measure, and (ii) the set S can be extended to a larger clonable set generated by a finite group G of order |G|=N, the number of states in the larger set. It is then shown that any local cloning apparatus is capable of cloning a number of states that divides D exactly. We provide a complete solution for two central problems in local cloning, giving necessary and sufficient conditions for (i) when a set of maximally entangled states can be locally cloned, valid for all D; and (ii) local cloning of entangled qubit states with nonvanishing entanglement. In both of these cases, we show that a maximally entangled resource is necessary and sufficient, and the states must be related to each other by local unitary “shift” operations. These shifts are determined by the group structure, so need not be simple cyclic permutations. Assuming this shifted form and partially entangled states, then in D=3 we show that a maximally entangled resource is again necessary and sufficient, while for higher-dimensional systems, we find that the resource state must be strictly more entangled than the states in S. All of our necessary conditions for separable operations are also necessary conditions for local operations and classical communication (LOCC), since the latter is a proper subset of the former. In fact, all our results hold for LOCC, as our sufficient conditions are demonstrated for LOCC, directly.

  8. Local cloning of entangled states

    SciTech Connect

    Gheorghiu, Vlad; Yu Li; Cohen, Scott M.

    2010-08-15

    We investigate the conditions under which a set S of pure bipartite quantum states on a DxD system can be locally cloned deterministically by separable operations, when at least one of the states is full Schmidt rank. We allow for the possibility of cloning using a resource state that is less than maximally entangled. Our results include that: (i) all states in S must be full Schmidt rank and equally entangled under the G-concurrence measure, and (ii) the set S can be extended to a larger clonable set generated by a finite group G of order |G|=N, the number of states in the larger set. It is then shown that any local cloning apparatus is capable of cloning a number of states that divides D exactly. We provide a complete solution for two central problems in local cloning, giving necessary and sufficient conditions for (i) when a set of maximally entangled states can be locally cloned, valid for all D; and (ii) local cloning of entangled qubit states with nonvanishing entanglement. In both of these cases, we show that a maximally entangled resource is necessary and sufficient, and the states must be related to each other by local unitary 'shift' operations. These shifts are determined by the group structure, so need not be simple cyclic permutations. Assuming this shifted form and partially entangled states, then in D=3 we show that a maximally entangled resource is again necessary and sufficient, while for higher-dimensional systems, we find that the resource state must be strictly more entangled than the states in S. All of our necessary conditions for separable operations are also necessary conditions for local operations and classical communication (LOCC), since the latter is a proper subset of the former. In fact, all our results hold for LOCC, as our sufficient conditions are demonstrated for LOCC, directly.

  9. Regulation of GNRH production by estrogen and bone morphogenetic proteins in GT1-7 hypothalamic cells.

    PubMed

    Otani, Hiroyuki; Otsuka, Fumio; Takeda, Masaya; Mukai, Tomoyuki; Terasaka, Tomohiro; Miyoshi, Tomoko; Inagaki, Kenichi; Suzuki, Jiro; Ogura, Toshio; Lawson, Mark A; Makino, Hirofumi

    2009-10-01

    Recent studies have shown that bone morphogenetic proteins (BMPs) are important regulators in the pituitary-gonadal endocrine axis. We here investigated the effects of BMPs on GNRH production controlled by estrogen using murine GT1-7 hypothalamic neuron cells. GT1-7 cells expressed estrogen receptor alpha (ERalpha; ESR1 as listed in MGI Database), ERbeta (ESR2 as listed in MGI Database), BMP receptors, SMADs, and a binding protein follistatin. Treatment with BMP2 and BMP4 had no effect on Gnrh mRNA expression; however, BMP6 and BMP7 significantly increased Gnrh mRNA expression as well as GnRH production by GT1-7 cells. Notably, the reduction of Gnrh expression caused by estradiol (E(2)) was restored by cotreatment with BMP2 and BMP4, whereas it was not affected by BMP6 or BMP7. E(2) activated extracellular signal-regulated kinase (ERK) 1/2 and stress-activated protein kinase/c-Jun NH(2)-terminal kinase (SAPK/JNK) signaling but did not activate p38-mitogen-activated protein kinase (MAPK) signaling in GT1-7 cells. Inhibition of ERK1/ERK2 reversed the inhibitory effect of estrogen on Gnrh expression, whereas SAPK/JNK inhibition did not affect the E(2) actions. Expression levels of Eralpha and Erbeta were reduced by BMP2 and BMP4, but were increased by BMP6 and BMP7. Treatment with an ER antagonist inhibited the E(2) effects on Gnrh suppression including reduction of E(2)-induced ERK phosphorylation, suggesting the involvement of genomic ER actions in Gnrh suppression. BMP2 and BMP4 also suppressed estrogen-induced phosphorylation of ERK1/ERK2 and SAPK/JNK signaling, suggesting that BMP2 and BMP4 downregulate estrogen effects by attenuating ER-MAPK signaling. Considering that BMP6 and BMP7 increased the expression of alpha1E-subunit of R-type calcium channel (Cacna1e), which is critical for GNRH secretion, it is possible that BMP6 and BMP7 directly stimulate GNRH release by GT1-7 cells. Collectively, a newly uncovered interaction of BMPs and ER may be involved in

  10. Cloning and expression of a cDNA encoding a maize glutathione-S-transferase in E. coli.

    PubMed

    Moore, R E; Davies, M S; O'Connell, K M; Harding, E I; Wiegand, R C; Tiemeier, D C

    1986-09-25

    The isolation and characterization of a family of maize glutathione-S-transferases (GST's) has been described previously. These enzymes are designated GSTs I, II and III based on size, substrate specificity and responsiveness to safeners. GST III has been shown to act on the herbicide alachlor as well as the commonly used substrate 1-chloro-2,4-dinitrobenzene (CDNB). Clones were isolated from a maize cDNA library in lambda gt10. Three clones contained the entire coding region for GST III. The sequences of these clones were consistent with the known amino terminal GST III protein sequence. Moreover, expression of one of these clones in E. coli resulted in a GST activity as measured with both CDNB and alachlor, proving that at least one of the clones encodes an active GST III species. With the enzyme expressed in E. coli it will become possible to study enzyme structure-function relationships ex planta. While a number of different GST proteins are present in maize tissue the GST III gene is present in single or low copy in the genome.

  11. Cloning and expression of a cDNA encoding a maize glutathione-S-transferase in E. coli.

    PubMed Central

    Moore, R E; Davies, M S; O'Connell, K M; Harding, E I; Wiegand, R C; Tiemeier, D C

    1986-01-01

    The isolation and characterization of a family of maize glutathione-S-transferases (GST's) has been described previously. These enzymes are designated GSTs I, II and III based on size, substrate specificity and responsiveness to safeners. GST III has been shown to act on the herbicide alachlor as well as the commonly used substrate 1-chloro-2,4-dinitrobenzene (CDNB). Clones were isolated from a maize cDNA library in lambda gt10. Three clones contained the entire coding region for GST III. The sequences of these clones were consistent with the known amino terminal GST III protein sequence. Moreover, expression of one of these clones in E. coli resulted in a GST activity as measured with both CDNB and alachlor, proving that at least one of the clones encodes an active GST III species. With the enzyme expressed in E. coli it will become possible to study enzyme structure-function relationships ex planta. While a number of different GST proteins are present in maize tissue the GST III gene is present in single or low copy in the genome. Images PMID:3532034

  12. [Mystery and problems of cloning].

    PubMed

    Nikitin, V A

    2010-01-01

    The attention of investigators is attracted to the fact that, in spite of great efforts in mammalian cloning, advances that have been made in this area of research are not great, and cloned animals have developmental pathologies often incompatible with life and/or reproduction ability. It is yet not clear what technical or biological factors underlie this, and how they are connected or interact with each other, which is more realistic strategically. There is a great number of articles dealing with the influence of cloning with the nuclear transfer on genetic and epigenetic reprogramming of donor cells. At the same time we can see the practical absence of analytical investigations concerning the technology of cloning as such, its weak points, and possible sources of cellular trauma in the course of microsurgery of nuclear transfer or twinning. This article discusses step by step several nuclear transfer techniques and the methods of dividing early preimplanted embryos for twinning with the aim to reveal possible sources of cell damage during micromanipulation that may have negative influence on the development of cloned organisms. Several new author's technologies based on the study of cell biophysical characteristics are described, which allow one to avoid cellular trauma during manipulation and minimize the possibility of cell damage at any rate.

  13. [Cloning and law in Hungary].

    PubMed

    Julesz, Máté

    2015-03-01

    Reproductive human cloning is prohibited in Hungary, as in many other countries. Therapeutic human cloning is not prohibited, just like in many other countries. Stem cell therapy is also allowed. Article III, paragraph (3) of the Hungarian basic law (constitution) strictly forbids total human cloning. Article 1 of the Additional Protocol to the Oviedo Convention, on the Prohibition of Cloning Human Beings (1998) stipulates that any intervention seeking to create a human being genetically identical to another human being, whether living or dead, is prohibited. In Hungary, according to Article 174 of the Criminal Code, total human cloning constitutes a crime. Article 180, paragraph (3) of the Hungarian Act on Health declares that embryos shall not be brought about for research purposes; research shall be conducted only on embryos brought about for reproductive purposes when this is authorized by the persons entitled to decide upon its disposal, or when the embryo is damaged. Article 180, paragraph (5) of the Hungarian Act on Health stipulates that multiple individuals who genetically conform to one another shall not be brought about. According to Article 181, paragraph (1) of the Hungarian Act on Health, an embryo used for research shall be kept alive for not longer than 14 days, not counting the time it was frozen for storage and the time period of research.

  14. Improvement of cloning efficiency in minipigs using post-thawed donor cells treated with roscovitine.

    PubMed

    Hwang, Seongsoo; Oh, Keon Bong; Kwon, Dae-Jin; Ock, Sun-A; Lee, Jeong-Woong; Im, Gi-Sun; Lee, Sung-Soo; Lee, Kichoon; Park, Jin-Ki

    2013-11-01

    Massachusetts General Hospital miniature pigs (MGH minipigs) have been established for organ transplantation studies across the homozygous major histocompatibility complex, but cloning efficiency of MGH minipigs is extremely low. This study was designed to increase the productivity of MGH minipigs by nuclear transfer of post-thaw donor cells after 1 h co-incubation with roscovitine. The MGH minipig cells were genetically modified with GT KO (alpha1,3-galactosyltransferase knock-out) and hCD46 KI (human CD46 knock-in) and used as donor cells. The GT KO/hCD46 KI donor cells were cultured for either 3 days (control group) or 1 h after thawing with 15 μM roscovitine (experimental group) prior to the nuclear transfer. The relative percentage of the transgenic donor cells that entered into G0/G1 was 93.7 % (±2.54). This was different from the donor cells cultured for 1 h with the roscovitine-treated group (84.6 % ±4.6) (P < 0.05) and without roscovitine (78.6 % ±5.5) (P < 0.01), respectively. The pregnancy rate and delivery rate in the roscovitine group (8/12 and 6/8, respectively) were significantly higher (P < 0.01) than those in the control group (6/19 and 3/6, respectively). In the experimental group, 12 GT KO/hCD46 KI transgenic minipigs were successfully generated, and five minipigs among them survived for more than 6 months so far. The recipient-based individual cloning efficiency ranged from 0.74 to 2.54 %. In conclusion, gene-modified donor cells can be used for cloning of MGH minipigs if the cells are post-thawed and treated with roscovitine for 1 h prior to nuclear transfer.

  15. Cloning and genomic nucleotide sequence of the matrix attachment region binding protein from the halotolerant alga Dunaliella salina.

    PubMed

    Wang, Peng-Ju; Wang, Tian-Yun; Wang, Ya-Feng; Yang, Rui; Li, Zhao-Xi

    2013-07-01

    In our previous study, the sequence of a matrix attachment region binding protein (MBP) cDNA was cloned from the unicellular green alga Dunaliella salina. However, the nucleotide sequence of this gene has not been reported so far. In this paper, the nucleotide sequence of MBP was cloned and characterized, and its gene copy number was determined. The MBP nucleotide sequence is 5641 bp long, and interrupted by 12 introns ranging from 132 to 562 bp. All the introns in the D. salina MBP gene have orthodox splice sites, exhibiting GT at the 5' end and AG at the 3' end. Southern blot analysis showed that MBP only has one copy in the D. salina genome.

  16. Isolation and characterization of two cDNA clones of anaerobically induced lactate dehydrogenase from barley roots

    SciTech Connect

    Hondred, D.; Hanson, A.D. )

    1990-05-01

    In barley roots during hypoxia, five lactate dehydrogenase (LDH) isozymes accumulate with a concomitant increase in enzyme activity ({approximately}20-fold). These isozymes are thought to be tetramers resulting from the random association of the products of two Ldh loci. To investigate this system, cDNA clones of LDH have been isolated from a {lambda}gt11 library using antiserum raised against barley LDH purified {approximately}3,000-fold and using nucleic acid probes synthesized by the polymerase chain reaction. Two cDNA clones were obtained (1,305 and 1,166 bp). The deduced amino acid sequences of the two barley LDHs are 96% identical to each other, and 50% and 40% identical to vertebrate and bacterial LDHs, respectively. Northern blots identified a single mRNA band ({approximately}1.5 kb) whose level rose 8-fold during hypoxia.

  17. Cloning and expression of human tyrosine aminotransferase cDNA.

    PubMed

    Séralini, G E; Luu-Thé, V; Labrie, F

    1995-01-02

    Complementary DNA clones encoding human tyrosine aminotransferase (TAT) were isolated by screening a normal adult woman liver lambda gt11 library with rat TAT cDNA. The largest isolated cDNA is 2051 bp long (EMBL accession number X55675). This cDNA was subcloned downstream of the cytomegalovirus promoter in the pCMV vector for transfection into human cervical carcinoma HeLa cells. Expression of the TAT cDNA resulted in the synthesis of a protein with a molecular mass of approximately 50 kDa, as assessed by Western analysis, a value which is in close agreement with the predicted molecular weight of 50,399, for a deduced sequence of 454 amino acids. The expressed protein catalyzed specifically the conversion of L-[14C]tyrosine into p-[14C]hydroxyphenylpyruvate. The availability of a functional TAT cDNA provides a useful tool for detailed study of the structure-function relationship of the enzyme and its mutated derivatives.

  18. The topsy-turvy cloning law.

    PubMed

    Brassington, Iain; Oultram, Stuart

    2011-03-01

    In debates about human cloning, a distinction is frequently drawn between therapeutic and reproductive uses of the technology. Naturally enough, this distinction influences the way that the law is framed. The general consensus is that therapeutic cloning is less morally problematic than reproductive cloning--one can hold this position while holding that both are morally unacceptable--and the law frequently leaves the way open for some cloning for the sake of research into new therapeutic techniques while banning it for reproductive purposes. We claim that the position adopted by the law has things the wrong way around: if we accept a moral distinction between therapeutic and reproductive cloning, there are actually more reasons to be morally worried about therapeutic cloning than about reproductive cloning. If cloning is the proper object of legal scrutiny, then, we ought to make sure that we are scrutinising the right kind of clone.

  19. Human cloning and human dignity.

    PubMed

    Birnbacher, Dieter

    2005-03-01

    Judging from the official documents dealing with the moral and legal aspects of human reproductive cloning there seems to be a nearly worldwide consensus that reproductive cloning is incompatible with human dignity. The certainty of this judgement is, however, not matched by corresponding arguments. Is the incompatibility of reproductive with human dignity an ultimate moral intuition closed to further argument? The paper considers several ways by which the intuition might be connected with more familiar applications of the concept of human dignity, and argues that there is no such connection. It concludes that the central objections to human reproductive cloning are not objections relating to dignity but objections relating to risk, especially the risks imposed on children born in the course of testing the method's safety.

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

    SciTech Connect

    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.

  1. Genotyping-in-Thousands by sequencing (GT-seq): A cost effective SNP genotyping method based on custom amplicon sequencing.

    PubMed

    Campbell, Nathan R; Harmon, Stephanie A; Narum, Shawn R

    2015-07-01

    Genotyping-in-Thousands by sequencing (GT-seq) is a method that uses next-generation sequencing of multiplexed PCR products to generate genotypes from relatively small panels (50-500) of targeted single-nucleotide polymorphisms (SNPs) for thousands of individuals in a single Illumina HiSeq lane. This method uses only unlabelled oligos and PCR master mix in two thermal cycling steps for amplification of targeted SNP loci. During this process, sequencing adapters and dual barcode sequence tags are incorporated into the amplicons enabling thousands of individuals to be pooled into a single sequencing library. Post sequencing, reads from individual samples are split into individual files using their unique combination of barcode sequences. Genotyping is performed with a simple perl script which counts amplicon-specific sequences for each allele, and allele ratios are used to determine the genotypes. We demonstrate this technique by genotyping 2068 individual steelhead trout (Oncorhynchus mykiss) samples with a set of 192 SNP markers in a single library sequenced in a single Illumina HiSeq lane. Genotype data were 99.9% concordant to previously collected TaqMan(™) genotypes at the same 192 loci, but call rates were slightly lower with GT-seq (96.4%) relative to Taqman (99.0%). Of the 192 SNPs, 187 were genotyped in ≥90% of the individual samples and only 3 SNPs were genotyped in <70% of samples. This study demonstrates amplicon sequencing with GT-seq greatly reduces the cost of genotyping hundreds of targeted SNPs relative to existing methods by utilizing a simple library preparation method and massive efficiency of scale.

  2. Hypertriglyceridemia associated with the c.553G>T APOA5 SNP results from aberrant hetero-disulfide bond formation

    PubMed Central

    Sharma, Vineeta; Witkowski, Andrzej; Witkowska, H. Ewa; Dykstra, Andrew; Simonsen, Jens B.; Nelbach, Lisa; Beckstead, Jennifer A.; Pullinger, Clive R.; Kane, John P.; Malloy, Mary J.; Watson, Gordon; Forte, Trudy M.; Ryan, Robert O.

    2014-01-01

    Objective Apolipoprotein (apo) A-V is a low abundance plasma protein that modulates triacylglycerol (TG) homeostasis. Gene transfer studies were undertaken in apoa5 (−/−) mice to define the mechanism underlying the correlation between the single nucleotide polymorphism (SNP) c.553G>T in APOA5 and hypertriglyceridemia (HTG). Approach and Results Adeno-associated virus (AAV) 2/8 mediated gene transfer of wild type (WT) apoA-V induced a dramatic lowering of plasma TG in apoa5 (−/−) mice while AAV2/8-Gly162Cys apoA-V (corresponding to the c.553G>T SNP: rs2075291) had a modest effect. Characterization studies revealed that plasma levels of WT- and G162C apoA-V in transduced mice were similar and within the physiological range. Fractionation of plasma from mice transduced with AAV2/8-G162C apoA-V indicated that, unlike WT apoA-V, >50% of G162C apoA-V was recovered in the lipoprotein-free fraction. Non-reducing SDS-PAGE immunoblot analysis provided evidence that G162C apoA-V present in the lipoprotein-free fraction, but not that portion associated with lipoproteins, displayed altered electrophoretic mobility consistent with disulfide-linked hetero-dimer formation. Immunoprecipitation followed by liquid chromatography/mass spectrometry of human plasma from subjects homozygous for WT APOA5 and c.553G>T APOA5 revealed that G162C apoA-V forms adducts with extraneous plasma proteins including fibronectin, kininogen-1 and others. Conclusion Substitution of Cys for Gly at position 162 of mature apoA-V introduces a free cysteine that forms disulfide bonds with plasma proteins such that its lipoprotein binding and TG modulation functions are compromised. PMID:25127531

  3. [The balanced force and the GT-rotary technique in comparison with the non-instrumental technique (NIT)].

    PubMed

    Lussi, Adrian; Hotz, Meret; Stich, Hermann

    2004-01-01

    The aim of the study was to compare the cleansing effect of the latest modification of the non-instrumentation technique (NIT) to that of conventional instrumentation. The root curvature in 100 vital human molars was determined by a standardized X-ray procedure and the teeth were assigned to five groups with 20 teeth each with an equal distribution of the root curvature. The preparation methods were the Balanced Force technique and the GT Rotary technique. Each root was irrigated with 40 ml of 3% sodium hypochlorite. The other groups were irrigated by NIT during 2.5, 5 or 10 minutes, respectively. The remaining pulpal tissue was stained and the root canals were exposed longitudinally. The teeth were then evaluated using a microscope and an image analysis-system. The residual organic debris in the apical, middle and coronal sections of the root canals were assessed as a percentage of the corresponding total examined length. The cleansing effect of the NIT in the coronal and middle parts of the canal used for 5 and 10 minutes was significantly better (p < 0.05) compared to using the device for 2.5 minutes. The cleansing effect of the NIT in the coronal and middle parts of the canal used for 5 and 10 minutes was also significantly better (p < 0.05) compared to using the GT Rotary or Balanced Force techniques. Apically, the cleansing effect of the NIT used for 5 and 10 minutes and the GT Rotary technique was significantly better (p < 0.05) compared to using the Balanced Force technique or the NIT for 2.5 minutes. It was concluded that the cleansing effect of the latest modification of the Non-instrumentation Technology (NIT) was equivalent to or better than that of conventional instrumentation requiring significantly less time.

  4. Brain-specific expression of MAP2 detected using a cloned cDNA probe

    PubMed Central

    1986-01-01

    We describe the isolation of a set of overlapping cDNAs encoding mouse microtubule associated protein 2 (MAP2), using an anti-MAP antiserum to screen a mouse brain cDNA expression library cloned in bacteriophage lambda gt11. The authenticity of these clones was established by the following criteria: (a) three non-identical clones each expressing a MAP2 immunoreactive fusion protein were independently isolated from the expression library; each of these clones cross-hybridized at the nucleic acid level; (b) anti-MAP antiserum was affinity purified using nitrocellulose-bound fusion protein; these antibodies detected only MAP2 in an immunoblot experiment of whole brain microtubule protein; (c) a series of cDNA "walking" experiments was done so as to obtain a non-overlapping cloned fragment corresponding to a different part of the same mRNA molecule. Upon subcloning this non-overlapping fragment into plasmid expression vectors, a fusion protein was synthesized that was immunoreactive with an anti-MAP2 specific antiserum. Thus, a single contiguous cloned mRNA molecule encodes at least two MAP2-specific epitopes; (d) the cloned cDNA probes detect an mRNA species in mouse brain that is of a size (approximately 9 kb) consistent with the coding capacity required by a 250,000-D protein. The MAP2-specific cloned cDNA probes were used in RNA blot transfer experiments to assay for the presence of MAP2 mRNA in a variety of mouse tissues. Though brain contained abundant quantities of MAP2 mRNA, no corresponding sequences were detectable in RNA prepared from liver, kidney, spleen, stomach, or thymus. We conclude that the expression of MAP2 is brain-specific. Use of the MAP2 specific cDNA probes in genomic Southern blot transfer experiments showed the presence of a single gene encoding MAP2 in mouse. The microheterogeneity of MAP2 is therefore ascribable either to alternative splicing within a single gene, or to posttranslational modification(s), or both. Under conditions of low

  5. Evaluation of Cathode Air Flow Transients in a SOFC/GT Hybrid System Using Hardware in the Loop Simulation.

    PubMed

    Zhou, Nana; Yang, Chen; Tucker, David

    2015-02-01

    Thermal management in the fuel cell component of a direct fired solid oxide fuel cell gas turbine (SOFC/GT) hybrid power system can be improved by effective management and control of the cathode airflow. The disturbances of the cathode airflow were accomplished by diverting air around the fuel cell system through the manipulation of a hot-air bypass valve in open loop experiments, using a hardware-based simulation facility designed and built by the U.S. Department of Energy, National Energy Technology Laboratory (NETL). The dynamic responses of the fuel cell component and hardware component of the hybrid system were studied in this paper.

  6. Typological and dimensional approach at comparing the Giessen Test (GT) with the NEO-Five-Factor-Inventory (NEO-FFI)

    PubMed Central

    Roth, Marcus; Körner, Annett; Herzberg, Philipp Yorck

    2008-01-01

    Objectives: This article reports comparisons of the Giessen Test (GT) with the NEO-Five-Factor-Inventory (NEO-FFI) based on a dimensional as well as on a typological approach. Method: Data were collected from 1673 subjects (aged between 18 and 96 years) constituting a representative sample of the German population. Results: The results indicate only moderate agreement (ranging from .25 to .61) between the subscales of the two personality inventories. The correspondence seems to be somewhat higher, when the typological approach was used instead of the dimensional approach. Conclusions: The typological approach is less dependent on the underlying questionnaires and provides a useful extension of the dimensional approach. PMID:19742276

  7. GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMNET REPORT FOR THE PERIOD JULY 1 THROUGH JULY 31, 2003

    SciTech Connect

    SHENOY, A.S.

    2003-07-01

    A271 GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMNET REPORT FOR THE PERIOD JULY 1 THROUGH JULY 31, 2003. Petten has completed design of the irradiation test rig for the HFR-EU2 test and has completed design and machining of the H-451 graphite sleeves which will be used to contain the HFR-EU2 fuel compacts. A plan, entitled ''Screening Tests for Selection of VHTR Advanced Fuel,'' has been drafted and has completed internal review. This screening program plan is a major portion of the Development Plan for Advanced High Temperature Coated-Particle currently under preparation.

  8. Operational, control and protective system transient analyses of the closed-cycle GT-HTGR power plant

    SciTech Connect

    Openshaw, F.L.; Chan, T.W.

    1980-07-01

    This paper presents a description of the analyses of the control/protective system preliminary designs for the gas turbine high-temperature gas-cooled reactor (GT-HTGR) power plant. The control system is designed to regulate reactor power, control electric load and turbine speed, control the temperature of the helium delivered to the turbines, and control thermal transients experienced by the system components. In addition, it provides the required control programming for startup, shutdown, load ramp, and other expected operations. The control system also handles conditions imposed on the system during upset and emergency conditions such as loop trip, reactor trip, or electrical load rejection.

  9. Healthy ageing of cloned sheep

    PubMed Central

    Sinclair, K. D.; Corr, S. A.; Gutierrez, C. G.; Fisher, P. A.; Lee, J.-H.; Rathbone, A. J.; Choi, I.; Campbell, K. H. S.; Gardner, D. S.

    2016-01-01

    The health of cloned animals generated by somatic-cell nuclear transfer (SCNT) has been of concern since its inception; however, there are no detailed assessments of late-onset, non-communicable diseases. Here we report that SCNT has no obvious detrimental long-term health effects in a cohort of 13 cloned sheep. We perform musculoskeletal assessments, metabolic tests and blood pressure measurements in 13 aged (7–9 years old) cloned sheep, including four derived from the cell line that gave rise to Dolly. We also perform radiological examinations of all main joints, including the knees, the joint most affected by osteoarthritis in Dolly, and compare all health parameters to groups of 5-and 6-year-old sheep, and published reference ranges. Despite their advanced age, these clones are euglycaemic, insulin sensitive and normotensive. Importantly, we observe no clinical signs of degenerative joint disease apart from mild, or in one case moderate, osteoarthritis in some animals. Our study is the first to assess the long-term health outcomes of SCNT in large animals. PMID:27459299

  10. Clone Poems and the Microcomputer.

    ERIC Educational Resources Information Center

    Irizarry, Estelle

    1989-01-01

    Describes how students can use the computer to study and create clone poems (altering original Spanish-language poems by substituting words and expressions), and how students can gain a deeper appreciation of the original poem's poetic structure and semantics. (CB)

  11. Clone Poems and the Microcomputer.

    ERIC Educational Resources Information Center

    Irizarry, Estelle

    1989-01-01

    Describes how students can use the computer to study and create clone poems (altering original Spanish-language poems by substituting words and expressions), and how students can gain a deeper appreciation of the original poem's poetic structure and semantics. (CB)

  12. Human reproductive cloning: a conflict of liberties.

    PubMed

    Havstad, Joyce C

    2010-02-01

    Proponents of human reproductive cloning do not dispute that cloning may lead to violations of clones' right to self-determination, or that these violations could cause psychological harms. But they proceed with their endorsement of human reproductive cloning by dismissing these psychological harms, mainly in two ways. The first tactic is to point out that to commit the genetic fallacy is indeed a mistake; the second is to invoke Parfit's non-identity problem. The argument of this paper is that neither approach succeeds in removing our moral responsibility to consider and to prevent psychological harms to cloned individuals. In fact, the same commitment to personal liberty that generates the right to reproduce by means of cloning also creates the need to limit that right appropriately. Discussion of human reproductive cloning ought to involve a careful and balanced consideration of both the relevant aspects of personal liberty - the parents' right to reproductive freedom and the cloned child's right to self-determination.

  13. Energy Values of Nine Populus Clones

    Treesearch

    Terry F. Strong

    1980-01-01

    Compares calorific values for components of nine Populus clones. The components include stem wood, stem bark, and branches. Also compares calorific values for clones of balsam poplar and black cottonwood parentages.

  14. Cloning and characterization of a c-myc intron binding protein (MIBP1).

    PubMed

    Makino, R; Akiyama, K; Yasuda, J; Mashiyama, S; Honda, S; Sekiya, T; Hayashi, K

    1994-12-25

    The cDNA for a c-myc intron 1 binding protein 1 (MIBP1) in the rat was isolated from lambda gt11 and lambda ZAPII cDNA libraries. Sequencing of the cDNA clones revealed a long ORF which encoded a putative protein of 2437 amino acid residues. This protein has two widely separated zinc finger regions, each of which carries C2H2 motifs. When expressed in E. coli as a fusion protein, part of the MIBP1 showed sequence-specific binding to the target sequence, i.e., a 9-bp sequence in the rat c-myc intron 1. MIBP1 is most likely the rat counterpart of human MHC binding protein-2 (MBP-2/HIV-EP2), based on the 86% similarity in nucleotide sequence and 93% similarity in amno acid sequence. Northern blotting revealed a high level of MIBP1 mRNA in the brain.

  15. Isolation and characterization of two homologous cDNA clones from Torpedo electromotor neurons.

    PubMed

    Ngsee, J K; Scheller, R H

    1989-10-01

    Two homologous cDNA clones were isolated from a Torpedo california electric lobe lambda gt11 expression library using a polyclonal antiserum directed against proteins associated with synaptic vesicles. Northern blotting reveals an 8- to 9-kb transcript in the electric lobe and the spinal cord, but not in the brain or other non-neuronal tissues. Antibodies generated against a fusion protein synthesized in Escherichia coli reacted with a 85- to 90-kD species in the neurons of the electric lobe. The immunoreactivity is associated with microsomal membranes and can be extracted readily with high salt. Immunohistochemical studies demonstrated a sparse punctate staining pattern in the cell body which colocalized with a subpopulation of post-Golgi vesicles.

  16. Isolation and nucleotide sequence of a cDNA clone encoding rat mitochondrial malate dehydrogenase.

    PubMed Central

    Grant, P M; Tellam, J; May, V L; Strauss, A W

    1986-01-01

    We have determined the complete sequence of the rat mitochondrial malate dehydrogenase (mMDH) precursor derived from nucleotide sequence of the cDNA. A single synthetic oligodeoxynucleotide probe was used to screen a rat atrial cDNA library constructed in lambda gt10. A 1.2 kb full-length cDNA clone provided the first complete amino acid sequence of pre-mMDH. The 1014 nucleotide-long open reading frame encodes the 314 residue long mature mMDH protein and a 24 amino acid NH2-terminal extension which directs mitochondrial import and is cleaved from the precursor after import to generate mature mMDH. The amino acid composition of the transit peptide is polar and basic. The pre-mMDH transit peptide shows marked homology with those of two other enzymes targeted to the rat mitochondrial matrix. Images PMID:3755817

  17. Probabilistic cloning of three symmetric states

    SciTech Connect

    Jimenez, O.; Bergou, J.; Delgado, A.

    2010-12-15

    We study the probabilistic cloning of three symmetric states. These states are defined by a single complex quantity, the inner product among them. We show that three different probabilistic cloning machines are necessary to optimally clone all possible families of three symmetric states. We also show that the optimal cloning probability of generating M copies out of one original can be cast as the quotient between the success probability of unambiguously discriminating one and M copies of symmetric states.

  18. Phase-covariant quantum cloning of qudits

    SciTech Connect

    Fan Heng; Imai, Hiroshi; Matsumoto, Keiji; Wang, Xiang-Bin

    2003-02-01

    We study the phase-covariant quantum cloning machine for qudits, i.e., the input states in a d-level quantum system have complex coefficients with arbitrary phase but constant module. A cloning unitary transformation is proposed. After optimizing the fidelity between input state and single qudit reduced density operator of output state, we obtain the optimal fidelity for 1 to 2 phase-covariant quantum cloning of qudits and the corresponding cloning transformation.

  19. Reproductive cloning: past, present and future.

    PubMed

    Gurdon, J B

    2005-03-01

    This brief outline in reproductive cloning describes the background to these studies and then discusses successive aspects of the subject. These include abnormalities in cloned animals, therapeutic cloning and the ethics of this subject. A reference to further reading is provided.

  20. Economical phase-covariant cloning of qudits

    SciTech Connect

    Buscemi, Francesco; D'Ariano, Giacomo Mauro; Macchiavello, Chiara

    2005-04-01

    We derive the optimal N{yields}M phase-covariant quantum cloning for equatorial states in dimension d with M=kd+N, k integer. The cloning maps are optimal for both global and single-qudit fidelity. The map is achieved by an 'economical' cloning machine, which works without ancilla.

  1. Local cloning of arbitrarily entangled multipartite states

    SciTech Connect

    Kay, Alastair; Ericsson, Marie

    2006-01-15

    We examine the perfect cloning of nonlocal, orthogonal states using only local operations and classical communication. We provide a complete characterisation of the states that can be cloned under these restrictions, and their relation to distinguishability. We also consider the case of catalytic cloning, which we show provides no enhancement to the set of clonable states.

  2. The eNOS 894G/T gene polymorphism and its influence on early and long-term mortality after on-pump cardiac surgery

    PubMed Central

    2013-01-01

    Background The eNOS 894G/T polymorphism (GG, GT, and TT) is associated with cardiovascular mortality and may influence cardiovascular diseases as a genetic risk factor. Moreover, this polymorphism has an impact on intraoperative hemodynamics during cardiac surgery with cardiopulmonary bypass (CPB). In this study, we analyzed the influence of this gene polymorphism on early clinical outcome in patients who underwent cardiac surgery with CPB. Also, we performed a 5-year follow-up, assessing the impact of this polymorphism on long-term mortality. Method 500 patients who underwent cardiac surgery with CPB between 2006 and 2007 were included in this prospective single centre study. Genotyping for the eNOS gene polymorphism was performed by polymerase chain reaction amplification. Results Genotype distribution of 894G/T was: GG 50.2%; GT 42.2%; TT 7.8%. Cardiovascular risk factors were equally distributed between the different genotypes of the eNOS 894G/T polymorphism. No significant difference among the groups was shown regarding Euroscore, SAPS II and APACHE II. Perioperative characteristics were also not affected by the genotypes, except for the consumption of norepinephrine (p = 0.03) and amiodarone (p = 0.01) which was higher in the GT allele carrier. The early postoperative course was quite uniform across the genotypes, except for mean intensive care unit length of stay which was significantly prolonged in GT carriers (p = 0.001). The five-year follow-up was 100% complete and showed no significant differences regarding mortality between the groups. Conclusion Our results show that the eNOS 894G /T polymorphism is not associated with early and late clinical outcome after cardiac surgery. Thus, this polymorphism can actually not help to identify high risk groups in the heterogeneous population of individuals who undergo cardiac surgery with CPB. PMID:24161078

  3. A novel Tetra-primer ARMS-PCR based assay for genotyping SNP rs12303764(G/T) of human Unc-51 like kinase 1 gene.

    PubMed

    Randhawa, Rohit; Duseja, Ajay; Changotra, Harish

    2017-02-01

    Various case-control studies have shown association of single nucleotide polymorphism rs12303764(G/T) in ULK1 with crohn's disease. The techniques used in these studies were time consuming, complicated and require sophisticated/expensive instruments. Therefore, in order to overcome these problems, we have developed a new, rapid and cost effective Tetra-primer ARMS-PCR assay to genotype single nucleotide polymorphism rs12303764(G/T) of ULK1 gene. We manually designed allele specific primers. DNA fragment amplified using outer primers was sequenced to obtain samples with known genotypes (GG, GT and TT) for further use in the development of T-ARMS-PCR assay. Amplification conditions were optimized for parameters; annealing temperature, Taq DNA polymerase and primers. The developed T-ARMS-PCR assay was applied to genotype one hundred samples from healthy individuals. Genotyping results of 10 DNA samples from healthy individuals for rs12303764(G/T) by T-ARMS-PCR assay and sequencing were concordant. The newly developed assay was further applied to genotype samples from 100 healthy individuals of North Indian origin. Genotype frequencies were 9, 34 and 57 % for GG, GT and TT, respectively. Allele frequencies were 0.26 and 0.74 for G and T, respectively. The allele frequencies were in Hardy-Weinberg's equilibrium (p = 0.2443). T-ARMS-PCR assay developed in our laboratory for genotyping rs12303764 (G/T) of ULK1 gene is time saving and cost-effective as compared to the available methods. Furthermore, this is the first study reporting allelic and genotype frequencies of ULK1 rs12303764 (G/T) variants in North Indian population.

  4. Analysis of systematic errors of the ASM/RXTE monitor and GT-48 γ-ray telescope

    NASA Astrophysics Data System (ADS)

    Fidelis, V. V.

    2011-06-01

    The observational data concerning variations of light curves of supernovae remnants—the Crab Nebula, Cassiopeia A, Tycho Brahe, and pulsar Vela—over 14 days scale that may be attributed to systematic errors of the ASM/RXTE monitor are presented. The experimental systematic errors of the GT-48 γ-ray telescope in the mono mode of operation were also determined. For this the observational data of TeV J2032 + 4130 (Cyg γ-2, according to the Crimean version) were used and the stationary nature of its γ-ray emission was confirmed by long-term observations performed with HEGRA and MAGIC. The results of research allow us to draw the following conclusions: (1) light curves of supernovae remnants averaged for long observing periods have false statistically significant flux variations, (2) the level of systematic errors is proportional to the registered flux and decreases with increasing temporal scale of averaging, (3) the light curves of sources may be modulated by the year period, and (4) the systematic errors of the GT-48 γ-ray telescope, in the amount caused by observations in the mono mode and data processing with the stereo-algorithm come to 0.12 min-1.

  5. Human OCT2 variant c.808G>T confers protection effect against cisplatin-induced ototoxicity

    PubMed Central

    Lanvers-Kaminsky, Claudia; Sprowl, Jason A; Malath, Ingrid; Deuster, Dirk; Eveslage, Maria; Schlatter, Eberhard; Mathijssen, Ron HJ; Boos, Joachim; Jürgens, Heribert; am Zehnhoff-Dinnesen, Antionette G; Sparreboom, Alex; Ciarimboli, Giuliano

    2016-01-01

    Aim Assuming that genetic variants of the SLC22A2 and SLC31A1 transporter affect patients’ susceptibility to cisplatin-induced ototoxicity, we compared the distribution of 11 SLC22A2 variants and the SLC31A1 variant rs10981694 between patients with and without cisplatin-induced ototoxicity. Patients & methods Genotyping was performed in 64 pediatric patients and significant findings were re-evaluated in 66 adults. Results The SLC22A2 polymorphism rs316019 (c.808G>T; Ser270Ala) was significantly associated with protection from cisplatin-induced ototoxicity in the pediatric (p = 0.022) and the adult cohort (p = 0.048; both: Fisher’s exact test). This result was confirmed by multiple logistic regression analysis accounting for age which was identified as a relevant factor for ototoxicity as well (rs316019: OR [G/T vs G/G] = 0.12, p = 0.009; age: OR [per year]: 0.84, p = 0.02). Conclusion These results identified rs316019 as potential pharmacogenomic marker for cisplatin-induced ototoxicity and point to a critical role of SLC22A2 for cisplatin transport in humans and its contribution to the organ specific side effects of this drug. PMID:25823781

  6. Computational Assessment of the GT-MHR Graphite Core Support Structural Integrity in Air-Ingress Accident Condition

    SciTech Connect

    Jong B. Lim; Eung S. Kim; Chang H. Oh; Richard R. Schultz; David A. Petti

    2008-10-01

    The objective of this project was to perform stress analysis for graphite support structures of the General Atomics’ 600 MWth GT-MHR prismatic core design using ABAQUS ® (ver. 6.75) to assess their structural integrity in air-ingress accident conditions where the structure weakens over time due to oxidation damages. The graphite support structures of prismatic type GT-MHR was analyzed based on the change of temperature, burn-off and corrosion depth during the accident period predicted by GAMMA, a multi-dimensional gas multi-component mixture analysis code developed in the Republic of Korea (ROK)/United States (US) International –Nuclear Engineering Research Initiative (I-NERI) project. Both the loading and thermal stresses were analyzed, but the thermal stress was not significant, leaving the loading stress to be the major factor. The mechanical strengths are exceeded between 11 to 11.5 days after loss-of-coolant-accident (LOCA), corresponding to 5.5 to 6 days after the start of natural convection.

  7. Human OCT2 variant c.808G>T confers protection effect against cisplatin-induced ototoxicity.

    PubMed

    Lanvers-Kaminsky, Claudia; Sprowl, Jason A; Malath, Ingrid; Deuster, Dirk; Eveslage, Maria; Schlatter, Eberhard; Mathijssen, Ron Hj; Boos, Joachim; Jürgens, Heribert; Am Zehnhoff-Dinnesen, Antionette G; Sparreboom, Alex; Ciarimboli, Giuliano

    2015-01-01

    Assuming that genetic variants of the SLC22A2 and SLC31A1 transporter affect patients' susceptibility to cisplatin-induced ototoxicity, we compared the distribution of 11 SLC22A2 variants and the SLC31A1 variant rs10981694 between patients with and without cisplatin-induced ototoxicity. Genotyping was performed in 64 pediatric patients and significant findings were re-evaluated in 66 adults. The SLC22A2 polymorphism rs316019 (c.808G>T; Ser270Ala) was significantly associated with protection from cisplatin-induced ototoxicity in the pediatric (p = 0.022) and the adult cohort (p = 0.048; both: Fisher's exact test). This result was confirmed by multiple logistic regression analysis accounting for age which was identified as a relevant factor for ototoxicity as well (rs316019: OR [G/T vs G/G] = 0.12, p = 0.009; age: OR [per year]: 0.84, p = 0.02). These results identified rs316019 as potential pharmacogenomic marker for cisplatin-induced ototoxicity and point to a critical role of SLC22A2 for cisplatin transport in humans and its contribution to the organ specific side effects of this drug. Original submitted 17 September 2014; Revision submitted 19 December 2014.

  8. A DOS Primer for Librarians.

    ERIC Educational Resources Information Center

    Beecher, Henry

    1989-01-01

    Presents a basic orientation to the functions and capabilities of disk operating systems (DOS), aimed at the nontechnically oriented user of IBM personal computers and compatible microcomputers. Areas discussed include booting up, the use of floppy and hard disks, file storage and manipulation, and directories. Further readings are provided. (CLB)

  9. A DOS Primer for Librarians.

    ERIC Educational Resources Information Center

    Beecher, Henry

    1989-01-01

    Presents a basic orientation to the functions and capabilities of disk operating systems (DOS), aimed at the nontechnically oriented user of IBM personal computers and compatible microcomputers. Areas discussed include booting up, the use of floppy and hard disks, file storage and manipulation, and directories. Further readings are provided. (CLB)

  10. Molecular cloning, expression, and primary sequence of outer membrane protein P2 of Haemophilus influenzae type b.

    PubMed Central

    Munson, R; Tolan, R W

    1989-01-01

    The structural gene for the porin of Haemophilus influenzae type b, designated outer membrane protein P2, was cloned, and the DNA sequence was determined. An oligonucleotide probe generated by reverse translation of N-terminal amino acid sequence data from the purified protein was used to screen genomic DNA. The probe detected a single EcoRI fragment of approximately 1,700 base pairs which was cloned to lambda gt11 and then into M13 and partially sequenced. The derived amino acid sequence indicated that we had cloned the N-terminal portion of the P2 gene. An overlapping approximately 1,600-base-pair PvuII genomic fragment was cloned into M13, and the sequence of the remainder of the P2 gene was determined. The gene for P2 was then reconstructed under the control of the T7 promoter and expressed in Escherichia coli. The N-terminal sequence of the purified protein corresponds to residues 21 through 34 of the derived amino acid sequence. Thus, the protein is synthesized with a 20-amino-acid leader peptide. The Mr of the processed protein is 37,782, in good agreement with the estimate of 37,000 from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Images PMID:2535836

  11. Therapeutic and reproductive cloning: a critique.

    PubMed

    Bowring, Finn

    2004-01-01

    This article is a critical examination of the science and ethics of human cloning. It summarises the key scientific milestones in the development of nuclear transplantation, explains the importance of cloning to research into the medical potential of embryonic stem cells, and discusses the well-worn distinction between 'therapeutic' and 'reproductive' cloning. Suggesting that this distinction will be impossible to police, it goes on to consider the ethics of full human cloning. It is concluded that it represents an unacceptable form of parental despotism, and that the genetic engineering and cloning of future human beings will fracture the foundations of modern humanism.

  12. Predators induce cloning in echinoderm larvae.

    PubMed

    Vaughn, Dawn; Strathmann, Richard R

    2008-03-14

    Asexual propagation (cloning) is a widespread reproductive strategy of plants and animals. Although larval cloning is well documented in echinoderms, identified stimuli for cloning are limited to those associated with conditions favorable for growth and reproduction. Our research shows that larvae of the sand dollar Dendraster excentricus also clone in response to cues from predators. Predator-induced clones were smaller than uncloned larvae, suggesting an advantage against visual predators. Our results offer another ecological context for asexual reproduction: rapid size reduction as a defense.

  13. Optimal quantum cloning via spin networks

    SciTech Connect

    Chen Qing; Cheng Jianhua; Wang Kelin; Du Jiangfeng

    2006-09-15

    In this paper we demonstrate that optimal 1{yields}M phase-covariant cloning quantum cloning is available via free dynamical evolution of spin networks. By properly designing the network and the couplings between spins, we show that optimal 1{yields}M phase-covariant cloning can be achieved if the initial state is prepared as a specific symmetric state. Especially, when M is an odd number, the optimal phase-covariant cloning can be achieved without ancillas. Moreover, we demonstrate that the same framework is capable for optimal 1{yields}2 universal cloning.

  14. No-cloning theorem on quantum logics

    SciTech Connect

    Miyadera, Takayuki; Imai, Hideki

    2009-10-15

    This paper discusses the no-cloning theorem in a logicoalgebraic approach. In this approach, an orthoalgebra is considered as a general structure for propositions in a physical theory. We proved that an orthoalgebra admits cloning operation if and only if it is a Boolean algebra. That is, only classical theory admits the cloning of states. If unsharp propositions are to be included in the theory, then a notion of effect algebra is considered. We proved that an atomic Archimedean effect algebra admitting cloning operation is a Boolean algebra. This paper also presents a partial result, indicating a relation between the cloning on effect algebras and hidden variables.

  15. Clone DB: an integrated NCBI resource for clone-associated data

    PubMed Central

    Schneider, Valerie A.; Chen, Hsiu-Chuan; Clausen, Cliff; Meric, Peter A.; Zhou, Zhigang; Bouk, Nathan; Husain, Nora; Maglott, Donna R.; Church, Deanna M.

    2013-01-01

    The National Center for Biotechnology Information (NCBI) Clone DB (http://www.ncbi.nlm.nih.gov/clone/) is an integrated resource providing information about and facilitating access to clones, which serve as valuable research reagents in many fields, including genome sequencing and variation analysis. Clone DB represents an expansion and replacement of the former NCBI Clone Registry and has records for genomic and cell-based libraries and clones representing more than 100 different eukaryotic taxa. Records provide details of library construction, associated sequences, map positions and information about resource distribution. Clone DB is indexed in the NCBI Entrez system and can be queried by fields that include organism, clone name, gene name and sequence identifier. Whenever possible, genomic clones are mapped to reference assemblies and their map positions provided in clone records. Clones mapping to specific genomic regions can also be searched for using the NCBI Clone Finder tool, which accepts queries based on sequence coordinates or features such as gene or transcript names. Clone DB makes reports of library, clone and placement data on its FTP site available for download. With Clone DB, users now have available to them a centralized resource that provides them with the tools they will need to make use of these important research reagents. PMID:23193260

  16. Clone DB: an integrated NCBI resource for clone-associated data.

    PubMed

    Schneider, Valerie A; Chen, Hsiu-Chuan; Clausen, Cliff; Meric, Peter A; Zhou, Zhigang; Bouk, Nathan; Husain, Nora; Maglott, Donna R; Church, Deanna M

    2013-01-01

    The National Center for Biotechnology Information (NCBI) Clone DB (http://www.ncbi.nlm.nih.gov/clone/) is an integrated resource providing information about and facilitating access to clones, which serve as valuable research reagents in many fields, including genome sequencing and variation analysis. Clone DB represents an expansion and replacement of the former NCBI Clone Registry and has records for genomic and cell-based libraries and clones representing more than 100 different eukaryotic taxa. Records provide details of library construction, associated sequences, map positions and information about resource distribution. Clone DB is indexed in the NCBI Entrez system and can be queried by fields that include organism, clone name, gene name and sequence identifier. Whenever possible, genomic clones are mapped to reference assemblies and their map positions provided in clone records. Clones mapping to specific genomic regions can also be searched for using the NCBI Clone Finder tool, which accepts queries based on sequence coordinates or features such as gene or transcript names. Clone DB makes reports of library, clone and placement data on its FTP site available for download. With Clone DB, users now have available to them a centralized resource that provides them with the tools they will need to make use of these important research reagents.

  17. Method for cloning lymphoblastoid cells

    SciTech Connect

    Hammerling, U.; Kosinski, S.

    1989-02-14

    A method is described for increasing cloning frequency of human lymphocyte or lumphoblastoid cells which have been transformed with Epstein Barr virus comprising growing the transformed cells in a semi-solid agarose medium. A lower and an upper layer of agarose are used, the lower layer comprising fibroblasts suspended in the agarose layer and the upper layer comprising irradiated fibroblasts and the transformed cells suspended in the agarose layer wherein the upper agarose layer is added after the lower layer has gelled.

  18. Different Roles of DosS and DosT in the Hypoxic Adaptation of Mycobacteria▿

    PubMed Central

    Kim, Min-Ju; Park, Kwang-Jin; Ko, In-Jeong; Kim, Young Min; Oh, Jeong-Il

    2010-01-01

    The DosS (DevS) and DosT histidine kinases form a two-component system together with the DosR (DevR) response regulator in Mycobacterium tuberculosis. DosS and DosT, which have high sequence similarity to each other over the length of their amino acid sequences, contain two GAF domains (GAF-A and GAF-B) in their N-terminal sensory domains. Complementation tests in conjunction with phylogenetic analysis showed that DevS of Mycobacterium smegmatis is more closely related to DosT than DosS. We also demonstrated in vivo that DosS and DosT of M. tuberculosis play a differential role in hypoxic adaptation. DosT responds to a decrease in oxygen tension more sensitively and strongly than DosS, which might be attributable to their different autooxidation rates. The different responsiveness of DosS and DosT to hypoxia is due to the difference in their GAF-A domains accommodating the hemes. Multiple alignment analysis of the GAF-A domains of mycobacterial DosS (DosT) homologs and subsequent site-directed mutagenesis revealed that just one substitution of E87, D90, H97, L118, or T169 of DosS with the corresponding residue of DosT is sufficient to convert DosS to DosT with regard to the responsiveness to changes in oxygen tension. PMID:20675480

  19. Lentivirus-mediated RNAi knockdown of LMP2A inhibits the growth of the Epstein-Barr-associated gastric carcinoma cell line GT38 in vitro

    PubMed Central

    Wang, Fangjun; Chen, Weichang; Liu, Pengfei; Zhou, Jundong; Liu, Bingtuan; Ye, Wu; Wang, Wenping; Shen, Xiuyun

    2017-01-01

    In this study, lentivirus-mediated RNA interference (RNAi) was applied to inhibit latent membrane protein 2A (LMP2A) gene expression, in order to explore the effects of LMP2A silencing on the growth of an Epstein-Barr virus-associated gastric carcinoma (EBVaGC) cell line in vitro. Lentivirus-mediated RNAi technology was employed to specifically knock down the LMP2A gene in the EBV-positive gastric carcinoma cell line GT38. After infection, reverse transcription-quantitative polymerase chain reaction, western blotting, flow cytometry and colony formation assays were conducted to evaluate the expression of LMP2A and the biological behavior of the GT38 cell line in vitro. The results showed that the expression of the LMP2A gene was clearly downregulated in the infected cells, which indicated that a highly efficient and stable lentivirus vector was successfully constructed. In the GT38 cells in which the expression of LMP2A was downregulated, the proliferation and colony formation of the cells was significantly inhibited. In addition, it was found that the cell cycle of the GT38 cells was arrested in the G0/G1 phase and the apoptosis rate was increased. These results indicate that lentivirus-mediated RNAi knockdown of LMP2A inhibits the growth of the EBVaGC cell line GT38 in vitro, and suggests that LMP2A is a potential target for gene therapy in the treatment of EBVaGC. PMID:28123488

  20. Chloride conducting light activated channel GtACR2 can produce both cessation of firing and generation of action potentials in cortical neurons in response to light.

    PubMed

    Malyshev, A Y; Roshchin, M V; Smirnova, G R; Dolgikh, D A; Balaban, P M; Ostrovsky, M A

    2017-02-15

    Optogenetics is a powerful technique in neuroscience that provided a great success in studying the brain functions during the last decade. Progress of optogenetics crucially depends on development of new molecular tools. Light-activated cation-conducting channelrhodopsin2 was widely used for excitation of cells since the emergence of optogenetics. In 2015 a family of natural light activated chloride channels GtACR was identified which appeared to be a very promising tool for using in optogenetics experiments as a cell silencer. Here we examined properties of GtACR2 channel expressed in the rat layer 2/3 pyramidal neurons by means of in utero electroporation. We have found that despite strong inhibition the light stimulation of GtACR2-positive neurons can surprisingly lead to generation of action potentials, presumably initiated in the axonal terminals. Thus, when using the GtACR2 in optogenetics experiments, its ability to induce action potentials should be taken into account. Our results also open an interesting possibility of using the GtACR2 both as cell silencer and cell activator in the same experiment varying the pattern of light stimulation.

  1. Crystal Structure of Botulinum Neurotoxin Type a in Complex With the Cell Surface Co-Receptor GT1b-Insight Into the Toxin-Neuron Interaction

    SciTech Connect

    Stenmark, P.; Dupuy, J.; Inamura, A.; Kiso, M.; Stevens, R.C.

    2009-05-26

    Botulinum neurotoxins have a very high affinity and specificity for their target cells requiring two different co-receptors located on the neuronal cell surface. Different toxin serotypes have different protein receptors; yet, most share a common ganglioside co-receptor, GT1b. We determined the crystal structure of the botulinum neurotoxin serotype A binding domain (residues 873-1297) alone and in complex with a GT1b analog at 1.7 A and 1.6 A, respectively. The ganglioside GT1b forms several key hydrogen bonds to conserved residues and binds in a shallow groove lined by Tryptophan 1266. GT1b binding does not induce any large structural changes in the toxin; therefore, it is unlikely that allosteric effects play a major role in the dual receptor recognition. Together with the previously published structures of botulinum neurotoxin serotype B in complex with its protein co-receptor, we can now generate a detailed model of botulinum neurotoxin's interaction with the neuronal cell surface. The two branches of the GT1b polysaccharide, together with the protein receptor site, impose strict geometric constraints on the mode of interaction with the membrane surface and strongly support a model where one end of the 100 A long translocation domain helix bundle swing into contact with the membrane, initiating the membrane anchoring event.

  2. [Cloning: applications in humans 2. Ethical considerations].

    PubMed

    de Wert, G M; Geraedts, J P

    2001-05-01

    Reproductive cloning in adults/children evokes unfavourable reactions. Direct objections are that cloning is unnatural, that it affects human dignity and violates the individual's right to genetic uniqueness. Consequential objections concern unjustified health risks for the progeny, unjustified psychosocial risks for the clone child and the risk of cloning for eugenetic purposes. There is consensus that reproductive cloning of existing persons is unjustify as yet because of the health risks for the offspring. Reproductive cloning of embryos is possible by means of nucleus transplantation and of embryo splitting. The ethical analysis of reproductive cloning of embryos depends on the purposes and applications. At least some of the moral objections against cloning of adults/children are not or not completely applicable to reproductive cloning of embryos. Conditions to be put to reproductive cloning of embryos are efficacy, safety and, at least for the time being, avoidance of asynchrony in transferring identical embryos. The ethical aspects of its application in the context of genetical reproductive techniques must be evaluated separately. Therapeutic cloning may be acceptable if alternatives are lacking.

  3. [Cloning: applications in humans. II. Ethical considerations].

    PubMed

    de Wert, G M; Geraedts, J P

    2000-05-13

    Reproductive cloning in adults/children evokes unfavourable reactions. Direct objections are that cloning is unnatural, that it affects human dignity and violates the individual's right to genetic uniqueness. Consequential objections concern unjustified health risks for the progeny, unjustified psychosocial risks for the clone child and the risk of cloning for eugenetic purposes. There is consensus that reproductive cloning of existing persons is unjustifiable as yet because of the health risks for the offspring. Reproductive cloning of embryos is possible by means of nucleus transplantation and of embryo splitting. The ethical analysis of reproductive cloning of embryos depends on the purposes and applications. At least some of the moral objections against cloning of adults/children are not or not completely applicable to reproductive cloning of embryos. Conditions to be put to reproductive cloning of embryos are efficacy, safety and, at least for the time being, avoidance of asynchrony in transferring identical embryos. The ethical aspects of its application in the context of genetical reproductive techniques must be evaluated separately. Therapeutic cloning may be acceptable if alternatives are lacking.

  4. Cloning Expeditions: Risky but Rewarding

    PubMed Central

    2013-01-01

    In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine. PMID:24061478

  5. Cloning expeditions: risky but rewarding.

    PubMed

    Lodish, Harvey

    2013-12-01

    In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine.

  6. Cloning and sequencing of the medium-chain S-acyl fatty acid synthetase thioester hydrolase cDNA from rat mammary gland.

    PubMed Central

    Naggert, J; Williams, B; Cashman, D P; Smith, S

    1987-01-01

    cDNA clones coding for the medium-chain S-acyl fatty acid synthetase thioester hydrolase (thioesterase II) from rat mammary gland were identified in a bacteriophage lambda gt11 library and their nucleotide sequences were determined. The predicted coding region spans 263 amino acid residues and includes a sequence identical with that of a peptide derived from the enzyme active site. The rat thioesterase II cDNA sequence exhibits homology with that of a thioesterase found in duck uropygial glands. Images Fig. 3. PMID:3632637

  7. GT0 Explosion Sources for IMS Infrasound Calibration: Charge Design and Yield Estimation from Near-source Observations

    NASA Astrophysics Data System (ADS)

    Gitterman, Y.; Hofstetter, R.

    2014-03-01

    Three large-scale on-surface explosions were conducted by the Geophysical Institute of Israel (GII) at the Sayarim Military Range, Negev desert, Israel: about 82 tons of strong high explosives in August 2009, and two explosions of about 10 and 100 tons of ANFO explosives in January 2011. It was a collaborative effort between Israel, CTBTO, USA and several European countries, with the main goal to provide fully controlled ground truth (GT0) infrasound sources, monitored by extensive observations, for calibration of International Monitoring System (IMS) infrasound stations in Europe, Middle East and Asia. In all shots, the explosives were assembled like a pyramid/hemisphere on dry desert alluvium, with a complicated explosion design, different from the ideal homogenous hemisphere used in similar experiments in the past. Strong boosters and an upward charge detonation scheme were applied to provide more energy radiated to the atmosphere. Under these conditions the evaluation of the actual explosion yield, an important source parameter, is crucial for the GT0 calibration experiment. Audio-visual, air-shock and acoustic records were utilized for interpretation of observed unique blast effects, and for determination of blast wave parameters suited for yield estimation and the associated relationships. High-pressure gauges were deployed at 100-600 m to record air-blast properties, evaluate the efficiency of the charge design and energy generation, and provide a reliable estimation of the charge yield. The yield estimators, based on empirical scaled relations for well-known basic air-blast parameters—the peak pressure, impulse and positive phase duration, as well as on the crater dimensions and seismic magnitudes, were analyzed. A novel empirical scaled relationship for the little-known secondary shock delay was developed, consistent for broad ranges of ANFO charges and distances, which facilitates using this stable and reliable air-blast parameter as a new potential

  8. Detailed Characterization of AR Coatings on Si Solar Cells: A New Application of GT-FabScan 6000; Preprint

    SciTech Connect

    Sopori, B.; Butterfield, B.; Amieva, J.

    2004-08-01

    We have developed a new application of GT-FabScan for rapid mapping of AR coatings on Si solar cells. The system generates an image of the AR thickness and presents it in a color format using false colors. This measurement is made in less than 100 ms. The development of this application enables the system to generate thickness maps of the AR coating to determine the repeatability of the deposition system, as well as to ensure that downstream processing can be controlled. These data can also be used to determine the average thickness of the coating. Downstream processing is an important issue in current solar cell technology. This paper describes its importance to the PV industry and discusses the principles and method of this measurement.

  9. GT2_proyer_3: Unveiling the evolutionary paths of the most massive stars through the study of their ejected nebulae

    NASA Astrophysics Data System (ADS)

    Royer, P.

    2011-05-01

    Several important questions remain open regarding the latest stages of evolution of the most massive stars, in particular regarding the exact evolutionary paths between the various subtypes of O stars, LBVs and Wolf-Rayet stars, and the mass-loss history of these objects throughout their lives. In the framework of the MESS GTKP+GT1, we have obtained or will obtain PACS imaging of 9 massive star nebulae of various types (LBV, LBV candidate, OF/WN, Of?p, WR) and PACS spectroscopy of 4 of them. In this short follow-up proposal we want to obtain PACS line spectroscopy for 3 peculiar massive and evolved objects for which spectroscopy is lacking. In particular, these observations will allow to determine the elemental abundances in the nebulae as well as the mass of the neutral gas using the fine structure lines formed in the ionized gas and in the photo-dissociation region respectively.

  10. GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD MAY 1 THROUGH MAY 31, 2003

    SciTech Connect

    SHENOY, A.S.

    2003-05-01

    A271 GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD MAY 1 THROUGH MAY 31, 2003. Petten advised GA the start of the HFR-EU2 irradiation is being delayed until late July 2004. HFR-EU1 (pebble fuel) is also delayed until February/March 2004. The reason for the delays was implementation of new financial regulations at Petten that delayed the contracts for capsule fabrication. Review of the MHR-2 Fuel Product Specification was completed. Revision of the specification to incorporate the review results is in progress. Detailed test matrices have been drafted for capsule irradiation tests and for post-irradiation heating tests proposed for development and qualification of advanced coated-particle fuels capable of meeting anticipated VHTR fuel performance requirements.

  11. GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD JUNE 1 THROUGH JUNE 30, 2003

    SciTech Connect

    SHENOY, A.S.

    2003-06-01

    A271 GT-MHR COMMERCIALZATION STUDY TECHNICAL PROGRESS AND COST MANAGEMENT REPORT FOR THE PERIOD JUNE 1 THROUGH JUNE 30, 2003. Petten was provided with irradiation dimensional change data for both fuel compacts and H-451 graphite for design of the graphite sleeves that hold the fuel compacts to be irradiated in HFR-EU2. The Fuel Sample Product Specification for the Fuel Performance Irradiation Test Capsule MHR-2 was completed and approved. A Work Breakdown Structure was prepared for the development and qualification of advanced coated-particle fuels capable of meeting anticipated fuel performance requirements and work was initiated on preparation of schedules and a cost estimates for the test matrices.

  12. Correlation between the NPPB gene promoter c.-1298 G/T polymorphism site and pulse pressure in the Chinese Han population.

    PubMed

    Zeng, K; Wu, X D; Cai, H D; Gao, Y G; Li, G; Liu, Q C; Gao, F; Chen, J H; Lin, C Z

    2014-04-29

    The aim of this study was to investigate the correlation between the natriuretic peptide precursor B (NPPB) gene single nucleotide polymorphism (SNP) c.-1298 G/T and pulse pressure (PP) of the Chinese Han population and the association between genotype and clinical indicators of hypertension. Peripheral blood was collected from 180 unrelated patients with hypertension and 540 healthy volunteers (control group), and DNA was extracted to amplify the 5'-flanking region and 2 exons of the NPPB gene by polymerase chain reaction; the fragment was sequenced after purification. The clinical data of all subjects were recorded, the distribution of the NPPB gene c.-1298 G/T polymorphism was determined, and differences in clinical indicators between the two groups were evaluated. The mean arterial pressure PP, and creatinine levels were significantly higher in the hypertension group than in the control group (P<0.05), but no other clinical indicators differed between the groups. There were no significant differences in genotype frequency and distribution of the NPPB gene c.-1298 G/T polymorphism between the hypertension group and the control group (P>0.05); in the control group, the mean PP of individuals with the SNP c.-1298 GG genotype was greater than that of individuals with the GT+TT genotype (P<0.05). In conclusion, there was no significant correlation between the NPPB gene c.-1298 G/T polymorphism and the incidence of essential hypertension in the Han population; however, the PP of the SNP c.-1298 GG genotype was greater than that of the GT+TT genotype in the control group.

  13. Tumor Necrosis Factor (TNF) -308G>A, Nitric Oxide Synthase 3 (NOS3) +894G>T Polymorphisms and Migraine Risk: A Meta-Analysis.

    PubMed

    Chen, Min; Tang, Wenjing; Hou, Lei; Liu, Ruozhuo; Dong, Zhao; Han, Xun; Zhang, Xiaofei; Wan, Dongjun; Yu, Shengyuan

    2015-01-01

    Conflicting data have been reported on the association between tumor necrosis factor (TNF) -308G>A and nitric oxide synthase 3 (NOS3) +894G>T polymorphisms and migraine. We performed a meta-analysis of case-control studies to evaluate whether the TNF -308G>A and NOS3 +894G>T polymorphisms confer genetic susceptibility to migraine. We performed an updated meta-analysis for TNF -308G>A and a meta-analysis for NOS3 +894G>T based on studies published up to July 2014. We calculated study specific odds ratios (OR) and 95% confidence intervals (95% CI) assuming allele contrast, dominant model, recessive model, and co-dominant model as pooled effect estimates. Eleven studies in 6682 migraineurs and 22591 controls for TNF -308G>A and six studies in 1055 migraineurs and 877 controls for NOS3 +894G>T were included in the analysis. Neither indicated overall associations between gene polymorphisms and migraine risk. Subgroup analyses suggested that the "A" allele of the TNF -308G>A variant increases the risk of migraine among non-Caucasians (dominant model: pooled OR = 1.82; 95% CI 1.15 - 2.87). The risk of migraine with aura (MA) was increased among both Caucasians and non-Caucasians. Subgroup analyses suggested that the "T" allele of the NOS3 +894G>T variant increases the risk of migraine among non-Caucasians (co-dominant model: pooled OR = 2.10; 95% CI 1.14 - 3.88). Our findings appear to support the hypothesis that the TNF -308G>A polymorphism may act as a genetic susceptibility factor for migraine among non-Caucasians and that the NOS3 +894G>T polymorphism may modulate the risk of migraine among non-Caucasians.

  14. From deep sequencing to actual clones.

    PubMed

    D'Angelo, Sara; Kumar, Sandeep; Naranjo, Leslie; Ferrara, Fortunato; Kiss, Csaba; Bradbury, Andrew R M

    2014-10-01

    The application of deep sequencing to in vitro display technologies has been invaluable for the straightforward analysis of enriched clones. After sequencing in vitro selected populations, clones are binned into identical or similar groups and ordered by abundance, allowing identification of those that are most enriched. However, the greatest strength of deep sequencing is also its greatest weakness: clones are easily identified by their DNA sequences, but are not physically available for testing without a laborious multistep process involving several rounds of polymerization chain reaction (PCR), assembly and cloning. Here, using the isolation of antibody genes from a phage and yeast display selection as an example, we show the power of a rapid and simple inverse PCR-based method to easily isolate clones identified by deep sequencing. Once primers have been received, clone isolation can be carried out in a single day, rather than two days. Furthermore the reduced number of PCRs required will reduce PCR mutations correspondingly. We have observed a 100% success rate in amplifying clones with an abundance as low as 0.5% in a polyclonal population. This approach allows us to obtain full-length clones even when an incomplete sequence is available, and greatly simplifies the subcloning process. Moreover, rarer, but functional clones missed by traditional screening can be easily isolated using this method, and the approach can be extended to any selected library (scFv, cDNA, libraries based on scaffold proteins) where a unique sequence signature for the desired clones of interest is available.

  15. Agro-economic impact of cattle cloning.

    PubMed

    Faber, D C; Ferre, L B; Metzger, J; Robl, J M; Kasinathan, P

    2004-01-01

    The purpose of this paper is to review the economic and social implications of cloned cattle, their products, and their offspring as related to production agriculture. Cloning technology in cattle has several applications outside of traditional production agriculture. These applications can include bio-medical applications, such as the production of pharmaceuticals in the blood or milk of transgenic cattle. Cloning may also be useful in the production of research models. These models may or may not include genetic modifications. Uses in agriculture include many applications of the technology. These include making genetic copies of elite seed stock and prize winning show cattle. Other purposes may range from "insurance" to making copies of cattle that have sentimental value, similar to cloning of pets. Increased selection opportunities available with cloning may provide for improvement in genetic gain. The ultimate goal of cloning has often been envisioned as a system for producing quantity and uniformity of the perfect dairy cow. However, only if heritability were 100%, would clone mates have complete uniformity. Changes in the environment may have significant impact on the productivity and longevity of the resulting clones. Changes in consumer preferences and economic input costs may all change the definition of the perfect cow. The cost of producing such animals via cloning must be economically feasible to meet the intended applications. Present inefficiencies limit cloning opportunities to highly valued animals. Improvements are necessary to move the applications toward commercial application. Cloning has additional obstacles to conquer. Social and regulatory acceptance of cloning is paramount to its utilization in production agriculture. Regulatory acceptance will need to address the animal, its products, and its offspring. In summary, cloning is another tool in the animal biotechnology toolbox, which includes artificial insemination, sexing of semen, embryo

  16. Chronic exposure to KATP channel openers results in attenuated glucose sensing in hypothalamic GT1-7 neurons.

    PubMed

    Haythorne, Elizabeth; Hamilton, D Lee; Findlay, John A; Beall, Craig; McCrimmon, Rory J; Ashford, Michael L J

    2016-12-01

    Individuals with Type 1 diabetes (T1D) are often exposed to recurrent episodes of hypoglycaemia. This reduces hormonal and behavioural responses that normally counteract low glucose in order to maintain glucose homeostasis, with altered responsiveness of glucose sensing hypothalamic neurons implicated. Although the molecular mechanisms are unknown, pharmacological studies implicate hypothalamic ATP-sensitive potassium channel (KATP) activity, with KATP openers (KCOs) amplifying, through cell hyperpolarization, the response to hypoglycaemia. Although initial findings, using acute hypothalamic KCO delivery, in rats were promising, chronic exposure to the KCO NN414 worsened the responses to subsequent hypoglycaemic challenge. To investigate this further we used GT1-7 cells to explore how NN414 affected glucose-sensing behaviour, the metabolic response of cells to hypoglycaemia and KATP activity. GT1-7 cells exposed to 3 or 24 h NN414 exhibited an attenuated hyperpolarization to subsequent hypoglycaemic challenge or NN414, which correlated with diminished KATP activity. The reduced sensitivity to hypoglycaemia was apparent 24 h after NN414 removal, even though intrinsic KATP activity recovered. The NN414-modified glucose responsiveness was not associated with adaptations in glucose uptake, metabolism or oxidation. KATP inactivation by NN414 was prevented by the concurrent presence of tolbutamide, which maintains KATP closure. Single channel recordings indicate that NN414 alters KATP intrinsic gating inducing a stable closed or inactivated state. These data indicate that exposure of hypothalamic glucose sensing cells to chronic NN414 drives a sustained conformational change to KATP, probably by binding to SUR1, that results in loss of channel sensitivity to intrinsic metabolic factors such as MgADP and small molecule agonists.

  17. Human pyridoxal phosphatase. Molecular cloning, functional expression, and tissue distribution.

    PubMed

    Jang, Young Min; Kim, Dae Won; Kang, Tae-Cheon; Won, Moo Ho; Baek, Nam-In; Moon, Byung Jo; Choi, Soo Young; Kwon, Oh-Shin

    2003-12-12

    Pyridoxal phosphatase catalyzes the dephosphorylation of pyridoxal 5'-phosphate (PLP) and pyridoxine 5'-phosphate. A human brain cDNA clone was identified to the PLP phosphatase on the basis of peptide sequences obtained previously. The cDNA predicts a 296-amino acid protein with a calculated Mr of 31698. The open reading frame is encoded by two exons located on human chromosome 22q12.3, and the exon-intron junction contains the GT/AG consensus splice site. In addition, a full-length mouse PLP phosphatase cDNA of 1978 bp was also isolated. Mouse enzyme encodes a protein of 292 amino acids with Mr of 31512, and it is localized on chromosome 15.E1. Human and mouse PLP phosphatase share 93% identity in protein sequence. A BLAST search revealed the existence of putative proteins in organism ranging from bacteria to mammals. Catalytically active human PLP phosphatase was expressed in Escherichia coli, and characteristics of the recombinant enzyme were similar to those of erythrocyte enzyme. The recombinant enzyme displayed Km and kcat values for pyridoxal of 2.5 microM and 1.52 s(-1), respectively. Human PLP phosphatase mRNA is differentially expressed in a tissue-specific manner. A single mRNA transcript of 2.1 kb was detected in all human tissues examined and was highly abundant in the brain. Obtaining the molecular properties for the human PLP phosphatase may provide new direction for investigating metabolic pathway involving vitamin B6.

  18. Cloning of the 5' mRNA for the 230-kD bullous pemphigoid antigen by rapid amplification of cDNA ends.

    PubMed

    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.

  19. The cloning of T lymphocytes.

    PubMed

    Schwartz, R H

    1982-02-01

    A new era of cellular immunology is clearly at hand. It is now possible, with a little bit of effort, to isolate monoclonal populations of T cells specific for any given antigen. The implications o f this technological advance are enormous in terms of applications to basic research and clinical medicine. In this article the two basic approaches that have been used to clone T lymphocytes are outlined, the pros and cons of each technique discussed and examples are given of recent experiments which have exploited this technology to gain new insights into T-cell specificity.

  20. Cloning cattle: the methods in the madness.

    PubMed

    Oback, Björn; Wells, David N

    2007-01-01

    Somatic cell nuclear transfer (SCNT) is much more widely and efficiently practiced in cattle than in any other species, making this arguably the most important mammal cloned to date. While the initial objective behind cattle cloning was commercially driven--in particular to multiply genetically superior animals with desired phenotypic traits and to produce genetically modified animals-researchers have now started to use bovine SCNT as a tool to address diverse questions in developmental and cell biology. In this paper, we review current cattle cloning methodologies and their potential technical or biological pitfalls at any step of the procedure. In doing so, we focus on one methodological parameter, namely donor cell selection. We emphasize the impact of epigenetic and genetic differences between embryonic, germ, and somatic donor cell types on cloning efficiency. Lastly, we discuss adult phenotypes and fitness of cloned cattle and their offspring and illustrate some of the more imminent commercial cattle cloning applications.

  1. Unified universal quantum cloning machine and fidelities

    SciTech Connect

    Wang Yinan; Shi Handuo; Xiong Zhaoxi; Jing Li; Mu Liangzhu; Ren Xijun; Fan Heng

    2011-09-15

    We present a unified universal quantum cloning machine, which combines several different existing universal cloning machines together, including the asymmetric case. In this unified framework, the identical pure states are projected equally into each copy initially constituted by input and one half of the maximally entangled states. We show explicitly that the output states of those universal cloning machines are the same. One importance of this unified cloning machine is that the cloning procession is always the symmetric projection, which reduces dramatically the difficulties for implementation. Also, it is found that this unified cloning machine can be directly modified to the general asymmetric case. Besides the global fidelity and the single-copy fidelity, we also present all possible arbitrary-copy fidelities.

  2. Therapeutic cloning research and ethical oversight.

    PubMed

    Spriggs, M

    2003-08-01

    Cloning Trevor, a story about therapeutic cloning research, appeared in the June issue of The Atlantic Monthly. The story gives a human face to the people whom therapeutic cloning could benefit. It presents an argument for government funding and it puts the usual calls for a moratorium on embryonic stem cell research to allow for more debate, in a less favourable light. The story also highlights some problems with ethical oversight.

  3. Telomeres and the ethics of human cloning.

    PubMed

    Allhoff, Fritz

    2004-01-01

    In search of a potential problem with cloning, I investigate the phenomenon of telomere shortening which is caused by cell replication; clones created from somatic cells will have shortened telomeres and therefore reach a state of senescence more rapidly. While genetic intervention might fix this problem at some point in the future, I ask whether, absent technological advances, this biological phenomenon undermines the moral permissibility of cloning.

  4. Structural features of the core proteins of human airway mucins ascertained by cDNA cloning.

    PubMed

    Porchet, N; Dufosse, J; Audie, J P; Duperat, V G; Perini, J M; Nguyen, V C; Degand, P; Aubert, J P

    1991-09-01

    Tracheobronchial secretions are one of the most important elements of the mucociliary system that protects the respiratory mucosa. They contain bronchial mucus, which is composed of a group of macromolecules secreted by the goblet cells of the epithelium and the submucosal glands. Bronchial mucins are the most characteristic molecules of this mucus. They form a group of complex, polydispersed O-linked glycoproteins containing sugars, which make up 80% of their weight. The protein core of human airway mucin has been difficult to sequence by traditional technologies because of its high content of serine and threonine residues linked to numerous oligosaccharide chains. We therefore prepared a lambda gt11 cDNA library from one sample of human tracheobronchial mucosa and screened this library with a polyclonal antibody directed against the apopeptides of human bronchial mucins. We obtained 20 positive clones that were sequenced. These sequences were classified into three different types. The use of the nucleotide probes from these clones in Northern blot analysis showed that the RNA messages were extremely polydispersed. At the current time, four of these probes allow us to map human tracheobronchial mucins genes to at least three different chromosomes. These results suggest that the peptide moiety of the human airway mucin is very heterogeneous.

  5. Human type VII collagen: cDNA cloning and chromosomal mapping of the gene

    SciTech Connect

    Parente, M.G.; Chung, L.C.; Ryynaenen, J.; Monli Chu; Uitto, J. ); Woodley, D.T.; Wynn, K.C.; Bauer, E.A. ); Mattei, M.G. )

    1991-08-15

    A human keratinocyte cDNA expression library in bacteriophage {lambda}gt11 was screened with the purified IgG fraction of serum from a patient with epidermolysis bullosa acquisita, which had a high titer of anti-type VII collagen antibodies. Screening of {approx}3 {times} 10{sup 5} plaques identified 8 positive clones, the largest one (K-131) being {approx}1.9 kilobases in size. Dideoxynucleotide sequencing of K-131 indicated that it consisted of 1875 base pairs and contained an open reading frame coding for a putative N-terminal noncollagenous domain of 439 amino acids and a collagenous domain was characterized by repeating Gly-Xaa-Yaa sequences that were interrupted in several positions by insertions or deletions of 1-3 amino acids. The deduced amino acid sequence also revealed a peptide segment that had a high degree of identity with a published type VII collagen protein sequence. The results mapped the COL7A1 to the locus 3p21. The cDNA clones characterized in this study will be valuable for understanding the protein structure and gene expression of type VII collagen present in anchoring fibrils and its aberrations in the dystrophic forms of heritable epidermolysis bullosa.

  6. Cloning and sequence analysis of the muramidase-2 gene from Enterococcus hirae.

    PubMed Central

    Chu, C P; Kariyama, R; Daneo-Moore, L; Shockman, G D

    1992-01-01

    Extracellular muramidase-2 of Enterococcus hirae ATCC 9790 was purified to homogeneity by substrate binding, guanidine-HCl extraction, and reversed-phase chromatography. A monoclonal antibody, 2F8, which specifically recognizes muramidase-2, was used to screen a genomic library of E. hirae ATCC 9790 DNA in bacteriophage lambda gt11. A positive phage clone containing a 4.5-kb DNA insert was isolated and analyzed. The EcoRI-digested 4.5-kb fragment was cut into 2.3-, 1.0-, and 1.5-kb pieces by using restriction enzymes KpnI, Sau3AI, and PstI, and each fragment was subcloned into plasmid pJDC9 or pUC19. The nucleotide sequence of each subclone was determined. The sequence data indicated an open reading frame encoding a polypeptide of 666 amino acid residues, with a calculated molecular mass of 70,678 Da. The first 24 N-terminal amino acids of purified extracellular muramidase-2 were in very good agreement with the deduced amino acid sequence after a 49-amino-acid putative signal sequence. Analysis of the deduced amino acid sequence showed the presence at the C-terminal region of the protein of six highly homologous repeat units separated by nonhomologous intervening sequences that are highly enriched in serine and threonine. The overall sequence showed a high degree of homology with a recently cloned Streptococcus faecalis autolysin. Images PMID:1347040

  7. Cloning and Characterization of a Critical Regulator for Preharvest Sprouting in Wheat

    PubMed Central

    Liu, Shubing; Sehgal, Sunish K.; Li, Jiarui; Lin, Meng; Trick, Harold N.; Yu, Jianming; Gill, Bikram S.; Bai, Guihua

    2013-01-01

    Sprouting of grains in mature spikes before harvest is a major problem in wheat (Triticum aestivum) production worldwide. We cloned and characterized a gene underlying a wheat quantitative trait locus (QTL) on the short arm of chromosome 3A for preharvest sprouting (PHS) resistance in white wheat using comparative mapping and map-based cloning. This gene, designated TaPHS1, is a wheat homolog of a MOTHER OF FLOWERING TIME (TaMFT)-like gene. RNA interference-mediated knockdown of the gene confirmed that TaPHS1 positively regulates PHS resistance. We discovered two causal mutations in TaPHS1 that jointly altered PHS resistance in wheat. One GT-to-AT mutation generates a mis-splicing site, and the other A-to-T mutation creates a premature stop codon that results in a truncated nonfunctional transcript. Association analysis of a set of wheat cultivars validated the role of the two mutations on PHS resistance. The molecular characterization of TaPHS1 is significant for expediting breeding for PHS resistance to protect grain yield and quality in wheat production. PMID:23821595

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

    SciTech Connect

    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. No end in sight to cloning debate.

    PubMed

    Graumann, Sigrid; Poltermann, Andreas

    2005-01-01

    Since last August, Great Britain has allowed the cloning for research purposes. This fact has re-generated an existing debate, taking into account the prohibition of cloning of the UN, the States are debating whether cloning should be prohibited or in the contrary, it should also be admitted for reproductive purposes. This situation has generated an international uneasiness due to the lack of a universal consensus. This article analyses this situation, bringing the reader closer to the very controversial texts, such as the European Constitution and the UN Convention on Cloning.

  10. Metabolomic phenotyping of a cloned pig model

    PubMed Central

    2011-01-01

    Background Pigs are widely used as models for human physiological changes in intervention studies, because of the close resemblance between human and porcine physiology and the high degree of experimental control when using an animal model. Cloned animals have, in principle, identical genotypes and possibly also phenotypes and this offer an extra level of experimental control which could possibly make them a desirable tool for intervention studies. Therefore, in the present study, we address how phenotype and phenotypic variation is affected by cloning, through comparison of cloned pigs and normal outbred pigs. Results The metabolic phenotype of cloned pigs (n = 5) was for the first time elucidated by nuclear magnetic resonance (NMR)-based metabolomic analysis of multiple bio-fluids including plasma, bile and urine. The metabolic phenotype of the cloned pigs was compared with normal outbred pigs (n = 6) by multivariate data analysis, which revealed differences in the metabolic phenotypes. Plasma lactate was higher for cloned vs control pigs, while multiple metabolites were altered in the bile. However a lower inter-individual variability for cloned pigs compared with control pigs could not be established. Conclusions From the present study we conclude that cloned and normal outbred pigs are phenotypically different. However, it cannot be concluded that the use of cloned animals will reduce the inter-individual variation in intervention studies, though this is based on a limited number of animals. PMID:21859467

  11. Quantum cloning disturbed by thermal Davies environment

    NASA Astrophysics Data System (ADS)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-06-01

    A network of quantum gates designed to implement universal quantum cloning machine is studied. We analyze how thermal environment coupled to auxiliary qubits, `blank paper' and `toner' required at the preparation stage of copying, modifies an output fidelity of the cloner. Thermal environment is described in terms of the Markovian Davies theory. We show that such a cloning machine is not universal any more but its output is independent of at least a part of parameters of the environment. As a case study, we consider cloning of states in a six-state cryptography's protocol. We also briefly discuss cloning of arbitrary input states.

  12. Species-specific challenges in dog cloning.

    PubMed

    Kim, G A; Oh, H J; Park, J E; Kim, M J; Park, E J; Jo, Y K; Jang, G; Kim, M K; Kim, H J; Lee, B C

    2012-12-01

    Somatic cell nuclear transfer (SCNT) is now an established procedure used in cloning of several species. SCNT in dogs involves multiple steps including the removal of the nuclear material, injection of a donor cell, fusion, activation of the reconstructed oocytes and finally transfer to a synchronized female recipient. There are therefore many factors that contribute to cloning efficiency. By performing a retrospective analysis of 2005-2012 published papers regarding dog cloning, we define the optimum procedure and summarize the specific feature for dog cloning.

  13. Human cloning: Eastern Mediterranean Region perspective.

    PubMed

    Abdur Rab, M; Khayat, M H

    2006-01-01

    Recent advances in genomics and biotechnology have ushered in a new era in health development. Therapeutic cloning possesses enormous potential for revolutionizing medical and therapeutic techniques. Cloning technology, however, is perceived as having the potential for reproductive cloning, which raises serious ethical and moral concerns. It is important that the Islamic countries come to a consensus on this vital issue. Developing science and technology for better health is a religious and moral obligation. There is an urgent need for Muslim scholars to discuss the issue of stem cell research and cloning rationally; such dialogue will not only consider the scientific merits but also the moral, ethical and legal implications.

  14. STRU-cloning: a fast, inexpensive and efficient cloning procedure applicable to both small scale and structural genomics size cloning.

    PubMed

    Bellini, Dom; Fordham-Skelton, Anthony P; Papiz, Miroslav Z

    2011-05-01

    We have developed a Single-Tube Restriction-based Ultrafiltration (STRU) cloning procedure that updates traditional ligation-dependent cloning to challenge the newer, faster and more efficient ligation-free techniques and could make it the method of choice. STRU-cloning employs centrifugal filter units with membrane of suitable cut off to remove small unwanted DNA fragments created during restriction of plasmids or PCR products. Heat inactivation, of restriction enzymes, followed by DNA ligation is then performed on the filtrate. By removing the agarose gel electrophoresis DNA purification step from the traditional protocol, which is time consuming and is known to be the cause of ligation problems, STRU-cloning becomes fast, very efficient, inexpensive and offers the highest degree of cloning flexibility by using restriction sites and can be performed in a single tube. This novel agarose gel-free cloning procedure provides benefits for both small and large scale cloning projects. Unlike traditional cloning it can be easily implemented as a fully automated process at very low costs.

  15. Metabolic Thresholds and Validated Accelerometer Cutoff Points for the Actigraph GT1M in Young Children Based on Measurements of Locomotion and Play Activities

    ERIC Educational Resources Information Center

    Jimmy, Gerda; Dossegger, Alain; Seiler, Roland; Mader, Urs

    2012-01-01

    The purpose of the current study was to determine metabolic thresholds and subsequent activity intensity cutoff points for the ActiGraph GT1M with various epochs spanning from 5 to 60 sec in young children. Twenty-two children, aged 4 to 9 years, performed 10 different activities including locomotion and play activities. Energy expenditure was…

  16. A cystic fibrosis patient homozygous for 621 + 1G-->T mutation has a severe pulmonary disease, mild pancreatic insufficiency and a gastro-esophageal reflux.

    PubMed

    Witt, M; Pogorzelski, A; Zebrak, J; Rutkiewicz, E

    1996-09-01

    A cystic fibrosis patient homozygous for 621 + 1G-->T mutation of the CFTR gene has been identified during a molecular screening program of Polish CF families. The patient is currently a 21-year-old female with severe pulmonary involvement, mild pancreatic insufficiency and complicated gastroesophageal reflux.

  17. Ameliorative effects of SLC22A2 gene polymorphism 808 G/T and cimetidine on cisplatin-induced nephrotoxicity in Chinese cancer patients.

    PubMed

    Zhang, Jing; Zhou, Wen

    2012-07-01

    To investigate the roles of SLC22A2 gene polymorphism 808 G/T and cimetidine on cisplatin-induced nephrotoxicity, a total of 123 Chinese cancer patients treated with cisplatin alone (n = 55) or in combination with cimetidine (n = 68) were genotyped. The changes of serum creatinine (SCr), blood urea nitrogen (BUN) and cystatin C levels were used as biomarkers for the evaluation of cisplatin-induced nephrotoxicity. The changes of BUN and SCr levels showed no significant difference between groups divided by genotypes and treatments (P > 0.05). However, patients with mutant genotype (GT/TT) or with cimetidine treatment had smaller increase of the cystatin C levels compared to those with wild genotype (GG) or without cimetidine treatment (P < 0.05). In the non-cimetidine-treated group, the changes of cystatin C level in patients with mutant genotype (GT/TT) was significantly smaller than those with wild genotype (GG) (P = 0.043). In the wild type group, the cystatin C level change of patients without cimetidine treatment was significantly larger than those with cimetidine treatment (P = 0.007). These results suggested that SLC22A2 gene polymorphism 808 G/T and cimetidine could attenuate cisplatin nephrotoxicity in Chinese cancer patients. But the renoprotection mechanism of cimetidine might be damaged by the mutation.

  18. Classification of Physical Activity Cut-Points and the Estimation of Energy Expenditure during Walking Using the GT3X+ Accelerometer in Overweight and Obese Adults

    ERIC Educational Resources Information Center

    Howe, Christopher C. F.; Moir, Hannah J.; Easton, Chris

    2017-01-01

    This study establishes tri-axial activity count (AC) cut-points for the GT3X+ accelerometer to classify physical activity intensity in overweight and obese adults. Further, we examined the accuracy of established and novel energy expenditure (EE) prediction equations based on AC and other metrics. "Part 1": Twenty overweight or obese…

  19. Sensitive detection of the c-KIT c.1430G>T mutation by mutant-specific polymerase chain reaction in feline mast cell tumours.

    PubMed

    Takanosu, M; Sato, M; Kagawa, Y

    2014-06-01

    Here, we describe the establishment of mutant-specific polymerase chain reaction (PCR) for detection of a c-KIT c.1430G>T mutation in feline mast cell tumours. Several mutations in feline c-KIT have been identified, with the c.1430G>T mutation accounting for a significant portion of feline mast cell tumour mutations. The c.1430G>T mutation in c-KIT exon 9 was detected in 15.7% (11 of 70) of samples by mutant-specific PCR but in only 7.1% (5 of 70) by PCR-restriction fragment length polymorphism (RFLP) in the genomic DNA isolated from 70 formalin-fixed paraffin-embedded sections or cells collected by fine needle aspiration. Mutant-specific PCR showed remarkably higher detection rate than did PCR-RFLP. DNA sequence analysis did not always yield identical results to those of mutant-specific PCR, suggesting heterogeneity of tumour cells. Mutant-specific PCR is a valid and efficient screening tool for detection of the c-KIT c.1430G>T point mutation in feline mast cell tumours compared with PCR-RFLP and sequencing analysis.

  20. Revisiting T-population derived from Tifrunner x GT-C20 for achieving high density genetic maps and detecting disease resistance QTLs in peanut (Arachis hypogaea L.)

    USDA-ARS?s Scientific Manuscript database

    Realizing the serious threats from Tomato spotted wilt virus (TSWV) and leaf spots (LS) to sustainable peanut production, high-density genetic linkage maps based on F2 and recombinant inbred line (RIL) T-population (Tifrunner × GT-C20) have been constructed and QTLs were identified in order to aid i...

  1. Health-based reference intervals for ALAT, ASAT and GT in serum, measured according to the recommendations of the European Committee for Clinical Laboratory Standards (ECCLS).

    PubMed

    Leino, A; Impivaara, O; Irjala, K; Mäki, J; Peltola, O; Järvisalo, J

    1995-05-01

    The reference intervals for the activities of L-alanine aminotransferase (EC 2.6.1.2, ALAT), L-aspartate aminotransferase (EC 2.6.1.1, ASAT) and gamma-glutamyltransferase (EC 2.3.2.2, GT) in serum were determined according to the recommendations of the European Committee for Clinical Laboratory Standards (ECCLS). Serum specimens from 954 subjects were analysed for ALAT and ASAT and from 794 subjects for GT. The subjects, aged 27-67 years, were participants in general health surveys. The reference population was formed by excluding subjects with any disease, or on any medication, affecting the liver, and also those consuming excessive amounts of alcohol. The 95% inner reference intervals for ALAT and ASAT were 9-50 (n = 189) and 15-36 U l-1 (n = 192) in men and 8-38 (n = 270) and 13-33 U l-1 (n = 270) in women. For GT the reference interval was 11-58 in men (n = 165) and 8-42 U l-1 in women (n = 220). Serum GT levels correlated clearly with alcohol consumption. Serum ALAT and ASAT were only slightly associated with alcohol consumption at levels less than 280 g per week in men and 190 g per week in women. There were modest positive associations between the three enzyme levels and body mass index. None of the enzymes correlated significantly with age.

  2. Endothelial nitric oxide synthase gene polymorphisms (promoter -786T/C, exon 894 G/T and intron G10T) in unexplained female infertility.

    PubMed

    Karatas, Ahmet; Eroz, Recep; Bahadır, Anzel; Keskin, Fatih; Ozlu, Tulay; Ozyalvaclı, Mehmet Emin

    2014-01-01

    Recent investigations in both males and females show that there may also be some genetic risk factors associated with infertility, and endothelial nitric oxide synthase (eNOS) has important functions in implantation. We aimed to investigate the association of three different polymorphisms of eNOS (promoter -786T/C, exon 894 G/T and intron G10T) with unexplained female infertility. Two groups of patients were included in the study: (1) women with unexplained infertility and (2) healthy, fertile women with normal menstrual cycles. eNOS polymorphisms were studied in genomic DNA of each patient by polymerase chain reaction-restriction fragment length polymorphism method. Forty-one women with unexplained infertility and 40 fertile women were included. Baseline physical characteristics and hormonal parameters of the two groups were similar. For eNOS exon 894 G/T polymorphism, the GG homozygotes were significantly lower and the heterozygotes GT were significantly higher in the infertile group than in the control group (p < 0.05). eNOS gene polymorphism both for promoter and intron were similar in the two groups (p > 0.05). Altered eNOS protein caused by eNOS exon 894 G/T polymorphism might cause implantation failure, which may be a possible cause of unexplained female infertility. © 2014 S. Karger AG, Basel.

  3. Draft Genome Sequences of Bifidobacterium angulatum GT102 and Bifidobacterium adolescentis 150: Focusing on the Genes Potentially Involved in the Gut-Brain Axis

    PubMed Central

    Dyachkova, Marina S.; Klimina, Ksenia M.; Kovtun, Alexey S.; Zakharevich, Natalia V.; Nezametdinova, Venera Z.; Averina, Olga V.

    2015-01-01

    The draft genome sequences of Bifidobacterium angulatum GT102 and Bifidobacterium adolescentis 150 strains isolated from the human intestinal microbiota are reported. Both strains are able to produce gamma-aminobutyric acid (GABA). Detailed genomes analysis will help to understand the role of GABA in the functioning of gut-brain axis. PMID:26139716

  4. Metabolic Thresholds and Validated Accelerometer Cutoff Points for the Actigraph GT1M in Young Children Based on Measurements of Locomotion and Play Activities

    ERIC Educational Resources Information Center

    Jimmy, Gerda; Dossegger, Alain; Seiler, Roland; Mader, Urs

    2012-01-01

    The purpose of the current study was to determine metabolic thresholds and subsequent activity intensity cutoff points for the ActiGraph GT1M with various epochs spanning from 5 to 60 sec in young children. Twenty-two children, aged 4 to 9 years, performed 10 different activities including locomotion and play activities. Energy expenditure was…

  5. Identification and characterisation of F3GT1 and F3GGT1, two glycosyltransferases responsible for anthocyanin biosynthesis in red-fleshed kiwifruit (Actinidia chinensis).

    PubMed

    Montefiori, Mirco; Espley, Richard V; Stevenson, David; Cooney, Janine; Datson, Paul M; Saiz, Anna; Atkinson, Ross G; Hellens, Roger P; Allan, Andrew C

    2011-01-01

    Much of the diversity of anthocyanins is due to the action of glycosyltransferases, which add sugar moieties to anthocyanidins. We identified two glycosyltransferases, F3GT1 and F3GGT1, from red-fleshed kiwifruit (Actinidia chinensis) that perform sequential glycosylation steps. Red-fleshed genotypes of kiwifruit accumulate anthocyanins mainly in the form of cyanidin 3-O-xylo-galactoside. Genes in the anthocyanin and flavonoid biosynthetic pathway were identified and shown to be expressed in fruit tissue. However, only the expression of the glycosyltransferase F3GT1 was correlated with anthocyanin accumulation in red tissues. Recombinant enzyme assays in vitro and in vivo RNA interference (RNAi) demonstrated the role of F3GT1 in the production of cyanidin 3-O-galactoside. F3GGT1 was shown to further glycosylate the sugar moiety of the anthocyanins. This second glycosylation can affect the solubility and stability of the pigments and modify their colour. We show that recombinant F3GGT1 can catalyse the addition of UDP-xylose to cyanidin 3-galactoside. While F3GGT1 is responsible for the end-product of the pathway, F3GT1 is likely to be the key enzyme regulating the accumulation of anthocyanin in red-fleshed kiwifruit varieties.

  6. Validity of the ActiGraph GT3X+ and BodyMedia SenseWear Armband to estimate energy expenditure during physical activity and sport.

    PubMed

    Gastin, Paul B; Cayzer, Cassy; Dwyer, Dan; Robertson, Sam

    2017-07-31

    The purpose of this study was to assess the validity of the ActiGraph GT3X+ (GT3X+) and the BodyMedia SenseWear Armband (SWA) to estimate energy expenditure (EE) during physical activity and field sport movements. Criterion validity. Twenty-six active adults completed a single 90min session involving alternating intervals of exercise (5min) and recovery (10min). Exercise involved walking (4km/h), jogging (8km/h), running (12km/h) or a sport-simulated circuit (three intervals). Participants wore two triaxial accelerometers (GT3X+ and SWA) and a portable gas analyser (MetaMax 3B), used as the criterion measure. Total EE was significantly underestimated (p<0.01) by the GT3X+ (mean bias±SD: -374.5±132.84kJ; % difference=-29.3%) and SWA (-244.3±148.0kJ; -18.2%). Overestimations were made by both accelerometers during the walk (GT3X+: 27.4±30.8kJ; SWA: 32.1±15.4kJ) and jog (38.0±30.0kJ; 34.5±31.6kJ). Underestimations were evident during the run (-41.2±25.1kJ; -43.8±33.5kJ) and circuit (C1: GTX+: -127.2±41.6kJ; SWA: -86.1±40.2kJ). Error of estimation increased in magnitude as the intensity of exercise increased (GT3X+: 40.8-143.0kJ; SWA: 35.5-102.0kJ). The ActiGraph GT3X+ and BodyMedia SWA do not provide valid EE estimates across a range of exercise modalities and intensities when compared to a criterion measure. Poor accuracy and large precision errors, particularly during high intensity and intermittent movement patterns, suggest these devices have limitations and should be used cautiously in the field. Copyright © 2017. Published by Elsevier Ltd.

  7. Role of Key Salt Bridges in Thermostability of G. thermodenitrificans EstGtA2: Distinctive Patterns within the New Bacterial Lipolytic Enzyme Family XV

    PubMed Central

    Charbonneau, David M.; Beauregard, Marc

    2013-01-01

    Bacterial lipolytic enzymes were originally classified into eight different families defined by Arpigny and Jaeger (families I-VIII). Recently, the discovery of new lipolytic enzymes allowed for extending the original classification to fourteen families (I-XIV). We previously reported that G. thermodenitrificans EstGtA2 (access no. AEN92268) belonged to a novel group of bacterial lipolytic enzymes. Here we propose a 15th family (family XV) and suggest criteria for the assignation of protein sequences to the N’ subfamily. Five selected salt bridges, hallmarks of the N’ subfamily (E3/R54, E12/R37, E66/R140, D124/K178 and D205/R220) were disrupted in EstGtA2 using a combinatorial alanine-scanning approach. A set of 14 (R/K→A) mutants was produced, including five single, three double, three triple and three quadruple mutants. Despite a high tolerance to non-conservative mutations for folding, all the alanine substitutions were destabilizing (decreasing Tm by 5 to 14°C). A particular combination of four substitutions exceeded this tolerance and prevents the correct folding of EstGtA2, leading to enzyme inactivation. Although other mutants remain active at low temperatures, the accumulation of more than two mutations had a dramatic impact on EstGtA2 activity at high temperatures suggesting an important role of these conserved salt bridge-forming residues in thermostability of lipolytic enzymes from the N’ subfamily. We also identified a particular interloop salt bridge in EstGtA2 (D194/H222), located at position i -2 and i -4 residues from the catalytic Asp and His respectively which is conserved in other related bacterial lipolytic enzymes (families IV and XIII) with high tolerance to mutations and charge reversal. We investigated the role of residue identity at position 222 in controlling stability-pH dependence in EstGtA2. The introduction of a His to Arg mutation led to increase thermostability under alkaline pH. Our results suggest primary targets for

  8. Molecular cloning and heterologous expression of novel glucosyltransferases from tobacco cultured cells that have broad substrate specificity and are induced by salicylic acid and auxin.

    PubMed

    Taguchi, G; Yazawa, T; Hayashida, N; Okazaki, M

    2001-07-01

    Scopoletin is one of the phytoalexins in tobacco. Cells of the T-13 cell line (Nicotiana tabacum L. Bright Yellow) accumulate a large amount of scopoletin, also known as 7-hydroxy-6-methoxycoumarin, as a glucoconjugate, scopolin, in vacuoles. We report here the molecular cloning of glucosyltransferases that can catalyze the glucosylation of many kinds of secondary metabolites including scopoletin. Two cDNAs encoding glucosyltransferase (NtGT1a and NtGT1b) were isolated from a cDNA library derived from the tobacco T-13 cell line by screening with heterologous cDNAs as a probe. The deduced amino-acid sequences of NtGT1a and NtGT1b exhibited 92% identity with each other, approximately 20-50% identities with other reported glucosyltransferases. Heterologous expression of these genes in Escherichia coli showed that the recombinant enzymes had glucosylation activity against both flavonoids and coumarins. They also strongly reacted with 2-naphthol as a substrate. These recombinant enzymes can utilize UDP-glucose as the sugar donor, but they can also utilize UDP-xylose as a weak donor. RNA blot analysis showed that these genes are induced by salicylic acid and auxin, but the time course of the expression was different. This result is similar to the changes in scopoletin glucosylation activity in these tobacco cells after addition of these plant growth regulators. These results might suggest that one of the roles of the products of these genes is scopoletin glucosylation, in response to salicylic acid and/or auxin, together with the other glucosyltransferases in tobacco cells.

  9. Positional cloning by linkage disequilibrium.

    PubMed

    Maniatis, Nikolas; Collins, Andrew; Gibson, Jane; Zhang, Weihua; Tapper, William; Morton, Newton E

    2004-05-01

    Recently, metric linkage disequilibrium (LD) maps that assign an LD unit (LDU) location for each marker have been developed (Maniatis et al. 2002). Here we present a multiple pairwise method for positional cloning by LD within a composite likelihood framework and investigate the operating characteristics of maps in physical units (kb) and LDU for two bodies of data (Daly et al. 2001; Jeffreys et al. 2001) on which current ideas of blocks are based. False-negative indications of a disease locus (type II error) were examined by selecting one single-nucleotide polymorphism (SNP) at a time as causal and taking its allelic count (0, 1, or 2, for the three genotypes) as a pseudophenotype, Y. By use of regression and correlation, association between every pseudophenotype and the allelic count of each SNP locus (X) was based on an adaptation of the Malecot model, which includes a parameter for location of the putative gene. By expressing locations in kb or LDU, greater power for localization was observed when the LDU map was fitted. The efficiency of the kb map, relative to the LDU map, to describe LD varied from a maximum of 0.87 to a minimum of 0.36, with a mean of 0.62. False-positive indications of a disease locus (type I error) were examined by simulating an unlinked causal SNP and the allele count was used as a pseudophenotype. The type I error was in good agreement with Wald's likelihood theorem for both metrics and all models that were tested. Unlike tests that select only the most significant marker, haplotype, or haploset, these methods are robust to large numbers of markers in a candidate region. Contrary to predictions from tagging SNPs that retain haplotype diversity, the sample with smaller size but greater SNP density gave less error. The locations of causal SNPs were estimated with the same precision in blocks and steps, suggesting that block definition may be less useful than anticipated for mapping a causal SNP. These results provide a guide to efficient

  10. cDNA cloning and analysis of betaine aldehyde dehydrogenase, a salt inducible enzyme in sugar beet

    SciTech Connect

    McCue, K.F.; Hanson, A.D. )

    1990-05-01

    Betaine accumulates and serves as a compatible osmolyte in some plants subjected to drought or salinity stress. The last enzyme in the betaine biosynthetic pathway is betaine aldehyde dehydrogenase (BADH). The activity of BADH increases in response to increasing salinity levels. This increase in activity corresponds to an increase in protein detectable by immunoblotting, and to an increase in the translatable BADH mRNA. BADH was cloned from a cDNA library constructed in {lambda}gt10 using poly(A){sup +} RNA from sugar beets salinized to 500 mM NaCl. cDNAs were size selected (>1kb) before ligation into the vector, and the library was screened with a spinach BADH cDNA probe. Three nearly full length clones obtained were confirmed as BADH by their nucleotide and deduced amino acid homology to spinach BADH. Clones averaged 1.8 kb and contained open reading frames of 500 amino acids at 80% identity with spinach BADH. RNA gel blot analysis of poly(A){sup +} RNA indicated that salinization to 500 mM NaCl resulted in a 5-fold increase of BADH mRNA level.

  11. Isolation of a complementary DNA clone for thyroid microsomal antigen. Homology with the gene for thyroid peroxidase.

    PubMed Central

    Seto, P; Hirayu, H; Magnusson, R P; Gestautas, J; Portmann, L; DeGroot, L J; Rapoport, B

    1987-01-01

    The thyroid microsomal antigen (MSA) in autoimmune thyroid disease is a protein of approximately 107 kD. We screened a human thyroid cDNA library constructed in the expression vector lambda gt11 with anti-107-kD monoclonal antibodies. Of five clones obtained, the recombinant beta-galactosidase fusion protein from one clone (PM-5) was confirmed to react with the monoclonal antiserum. The complementary DNA (cDNA) insert from PM-5 (0.8 kb) was used as a probe on Northern blot analysis to estimate the size of the mRNA coding for the MSA. The 2.9-kb messenger RNA (mRNA) species observed was the same size as that coding for human thyroid peroxidase (TPO). The probe did not bind to human liver mRNA, indicating the thyroid-specific nature of the PM-5-related mRNA. The nucleotide sequence of PM-5 (842 bp) was determined and consisted of a single open reading frame. Comparison of the nucleotide sequence of PM-5 with that presently available for pig TPO indicates 84% homology. In conclusion, a cDNA clone representing part of the microsomal antigen has been isolated. Sequence homology with porcine TPO, as well as identity in the size of the mRNA species for both the microsomal antigen and TPO, indicate that the microsomal antigen is, at least in part, TPO. Images PMID:3654979

  12. Evaluation of rs62527607 [GT] single nucleotide polymorphism located in BAALC gene in children with acute leukemia using mismatch PCR-RFLP.

    PubMed

    Nadimi, Motahareh; Rahgozar, Soheila; Moafi, Alireza; Tavassoli, Manoochehr; Mesrian Tanha, Hamzeh

    2016-01-01

    Acute leukemia is the most common cancer in children and involves several factors that contribute to the development of multidrug resistance and treatment failure. According to our recent studies, the BAALC gene is identified to have high mRNA expression levels in childhood acute lymphoblastic leukemia (ALL) and those with multidrug resistance. Several polymorphisms are associated with the expression of this gene. To date, there has been no study on the rs62527607 [GT] single nucleotide polymorphism (SNP) of BAALC gene and its link with childhood acute lymphoblastic and myeloid leukemia (AML). The purpose of this study is to evaluate the prevalence of this polymorphism in pediatric acute leukemia, as well as its relationship with prognosis. DNA samples were extracted from bone marrow slides of 129 children with ALL and 16 children with AML. The rs62527607 [GT] SNP was evaluated using mismatch polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based analysis. The association between the SNP alleles and patient disease-free survival was then assessed. The prevalence of the T-allele of rs62527607 [GT] SNP in childhood T-ALL and pre-B-ALL was 28.3% and 11.2%, respectively. In the pre-B-ALL patients, 3 year disease free survival was associated with the GG genotype. Results showed a robust association between the rs62527607 SNP and the risk of relapse in ALL, but not AML, patients. T-ALL patients with the GT genotype had an 8.75 fold higher risk of relapse. The current study demonstrates a significant association between the genotype GT and the polymorphic allele G424T, and introduces this SNP as a negative prognostic factor in children with ALL.

  13. GT-repeat polymorphism in the heme oxygenase-1 gene promoter is associated with cardiovascular mortality risk in an arsenic-exposed population in northeastern Taiwan

    SciTech Connect

    Wu, Meei-Maan; Chiou, Hung-Yi; Chen, Chi-Ling; Wang, Yuan-Hung; Hsieh, Yi-Chen; Lien, Li-Ming; Lee, Te-Chang; Chen, Chien-Jen

    2010-11-01

    Inorganic arsenic has been associated with increased risk of atherosclerotic vascular disease and mortality in humans. A functional GT-repeat polymorphism in the heme oxygenase-1 (HO-1) gene promoter is inversely correlated with the development of coronary artery disease and restenosis after clinical angioplasty. The relationship of HO-1 genotype with arsenic-associated cardiovascular disease has not been studied. In this study, we evaluated the relationship between the HO-1 GT-repeat polymorphism and cardiovascular mortality in an arsenic-exposed population. A total of 504 study participants were followed up for a median of 10.7 years for occurrence of cardiovascular deaths (coronary heart disease, cerebrovascular disease, and peripheral arterial disease). Cardiovascular risk factors and DNA samples for determination of HO-1 GT repeats were obtained at recruitment. GT repeats variants were grouped into the S (< 27 repeats) or L allele ({>=} 27 repeats). Relative mortality risk was estimated using Cox regression analysis, adjusted for competing risk of cancer and other causes. For the L/L, L/S, and S/S genotype groups, the crude mortalities for cardiovascular disease were 8.42, 3.10, and 2.85 cases/1000 person-years, respectively. After adjusting for conventional cardiovascular risk factors and competing risk of cancer and other causes, carriers with class S allele (L/S or S/S genotypes) had a significantly reduced risk of cardiovascular mortality compared to non-carriers (L/L genotype) [OR, 0.38; 95% CI, 0.16-0.90]. In contrast, no significant association was observed between HO-1 genotype and cancer mortality or mortality from other causes. Shorter (GT)n repeats in the HO-1 gene promoter may confer protective effects against cardiovascular mortality related to arsenic exposure.

  14. COL1A1 gene -1997G/T polymorphism and risk of osteoporosis in postmenopausal women: a meta-analysis.

    PubMed

    Yu, K H; Tang, J; Dai, C Q; Yu, Y; Hong, J J

    2015-09-21

    Studies investigating the association between the COL1A1 gene -1997G/T polymorphism and the risk of osteoporosis in postmenopausal women have reported conflicting results. We performed a meta-analysis based on the evidence currently available from the literature to make a more precise estimation of this relationship. We conducted searches of the published literature in the PubMed and Embase databases up to September 2014. We estimated the pooled odds ratios with their 95% confidence intervals to assess the associations using fixed- or random-effect models. Publication bias was investigated by Begg's funnel plot. Meta-analysis was performed using the STATA package version 12.0. No significant association was found between the -1997G/T polymorphism in the COL1A1 gene and osteoporosis risk in the total population analysis (TT vs GG: OR = 1.28, 95%CI = 0.76-2.17; TT vs GT: OR = 1.04, 95%CI = 0.60-1.78; dominant model: OR = 0.84, 95%CI = 0.50-1.40; recessive model: OR = 1.18, 95%CI = 0.84- 1.66). In a subgroup analysis by nationality, the results also showed that no significant associations between the COL1A1 gene -1997G/T polymorphism and osteoporosis risk existed in either Caucasian or Asian populations. No evidence of publication bias was found. In conclusion, the COL1A1 gene -1997G/T polymorphism might not be a risk factor for osteoporosis in postmenopausal women. Further large and well-designed studies are needed to confirm these conclusions.

  15. Association between +4259 T>G and -574 G>T Polymorphisms of TIM-3 with Asthma in an Iranian Population.

    PubMed

    Sadri, Maryam; Ganjalikhani-Hakemi, Mazdak; Akbari, Peyman; Salehi, Rasoul; Rastaghi, Sedighe; Ghasemi, Ramin; Meshkat, Rezvan

    2017-08-01

    T-cell immunoglobulin and mucin domain (TIM)-3 have been shown to negatively regulate Th1 cell-mediated immunity. Activation of TIM-3 by galectin-9 induces Th1 cell apoptosis, which may contribute to skewing of immune response towards Th2-dominant immunity. The aim of this study was to determine whether certain genetic variations of TIM-3 influence predisposition to asthma in a sample of Iranian population. This case-control study was conducted on 209 patients with asthma and 200 healthy controls. The +4259 T>G and -574 G>T polymorphisms were detected using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and amplification refractory mutation system-PCR(ARMS-PCR). Total serum IgE was further measured with ELISA. Notably, +4259T > G and-574G>T polymorphisms of TIM-3 were significantly associated with the susceptibility to asthma. In addition, the present study showed a significant difference between the distribution frequency of the GT + TT genotype and T allele on the +4259 T>G and -574 G>T locus between the groups.However, no correlation between the +4259 T > G and -574G > T polymorphisms and total serum IgE levels were observed. Together these results suggest that the TIM-3 +4259 T>G and -574 G>T polymorphisms are greatly associated with the susceptibility of Iranian population to asthma, which could open up new horizons for  better understanding of the pathophysiology, diagnostic, prognostic and therapeutic approaches of asthma.

  16. cDNA clones of the auxin-binding protein from corn coleoptiles (Zea mays L.): isolation and characterization by immunological methods.

    PubMed

    Tillmann, U; Viola, G; Kayser, B; Siemeister, G; Hesse, T; Palme, K; Löbler, M; Klämbt, D

    1989-09-01

    An auxin-binding protein (ABP) cDNA clone was selected from a lambda gt11 cDNA library from corn coleoptiles with highly purified IgGanti ABP. The sequence of 794 bp contains an open reading frame (ORF) of 603 bp, coding for a 22 kd protein. There are indications of a signal peptide of 38 amino acids (von Heijne, G. 1983, Eur. J. Biochem., 133, 17-21). A N-glycosylation site can be deduced and a C-terminal KDEL amino acid sequence is detected. An EcoRI fragment containing the beginning portion of the cDNA with about three quarters of the ORF was used to select cDNA clones from an independently produced lambda gt11 cDNA library of corn coleoptiles. Northern blot analysis with in vitro transcribed biotinylated RNA showed a single band of not more than 850 bases. The full-length in vitro transcript directed the in vitro synthesis of a protein which is precipitated by IgGanti ABP. Rabbit antibodies raised against a fusion protein detect the ABP as a double band on Western blots. Only the smaller of the two ABP bands is labeled by two different KDEL-specific IgG preparations.

  17. Recombination-assisted megaprimer (RAM) cloning

    PubMed Central

    Mathieu, Jacques; Alvarez, Emilia; Alvarez, Pedro J.J.

    2014-01-01

    No molecular cloning technique is considered universally reliable, and many suffer from being too laborious, complex, or expensive. Restriction-free cloning is among the simplest, most rapid, and cost-effective methods, but does not always provide successful results. We modified this method to enhance its success rate through the use of exponential amplification coupled with homologous end-joining. This new method, recombination-assisted megaprimer (RAM) cloning, significantly extends the application of restriction-free cloning, and allows efficient vector construction with much less time and effort when restriction-free cloning fails to provide satisfactory results. The following modifications were made to the protocol:•Limited number of PCR cycles for both megaprimer synthesis and the cloning reaction to reduce error propagation.•Elimination of phosphorylation and ligation steps previously reported for cloning methods that used exponential amplification, through the inclusion of a reverse primer in the cloning reaction with a 20 base pair region of homology to the forward primer.•The inclusion of 1 M betaine to enhance both reaction specificity and yield. PMID:26150930

  18. Reversibility of continuous-variable quantum cloning

    SciTech Connect

    Filip, Radim; Marek, Petr; Fiurasek, Jaromir

    2004-01-01

    We analyze a reversibility of optimal Gaussian 1{yields}2 quantum cloning of a coherent state using only local operations on the clones and classical communication between them and propose a feasible experimental test of this feature. Performing Bell-type homodyne measurement on one clone and anticlone, an arbitrary unknown input state (not only a coherent state) can be restored in the other clone by applying appropriate local unitary displacement operation. We generalize this concept to a partial reversal of the cloning using only local operations and classical communication (LOCC) and we show that this procedure converts the symmetric cloner to an asymmetric cloner. Further, we discuss a distributed LOCC reversal in optimal 1{yields}M Gaussian cloning of coherent states which transforms it to optimal 1{yields}M{sup '} cloning for M{sup '}cloning as a possible eavesdropping attack on quantum communication link, the reversibility can be utilized to improve the security of the link even after the attack.

  19. The ethics of human reproductive cloning.

    PubMed

    Strong, Carson

    2005-03-01

    This article addresses the question of whether human reproductive cloning could be ethically justifiable in at least some cases involving infertile couples who would choose cloning as a way to have a genetically related child. At present, the risk of congenital anomalies constitutes a compelling argument against human reproductive cloning. The article explores whether reproductive cloning could be ethically justifiable if, at some future time, cloning becomes possible without an elevated risk of anomalies. It is argued that freedom to use cloning is a form of procreative freedom and, as such, deserves respect. All of the objections that have been raised against human reproductive cloning fall under three main categories: those that appeal to the interests of the child, those based on consequences for society, and those arising from teleological views. Objections that appeal to the child's interests are, in turn, of two main kinds: consequentialist and deontological. All of these types of objections are examined, and it is found that each involves serious problems that prevent it from being a reasonable objection in the context of the infertility cases considered. It is concluded that human reproductive cloning would be ethically justifiable in at least some cases involving infertile couples, provided that it could be performed without an elevated risk of anomalies.

  20. Impact of cloning on cattle breeding systems.

    PubMed

    McClintock, A E

    1998-01-01

    The concept of clone-family testing is compared with existing progeny testing systems. The critical factors that will decide how cloning is utilized are the potential size of cloned families, and the cost per embryo (or per calf born). If family sizes of 100,000 become routinely achievable (cheaply), then clone testing becomes viable. In rough figures, cloned embryos costing $30 with a 50% calving rate would be attractive to farmers and would be cheap enough that farmers would buy more (crossbred) embryos in order to breed further replacement cows. At $300 per embryo, farmers would be more inclined to buy a number of cloned pure-bred female embryos and then to use conventional artificial insemination to breed further replacements from these superior cows. At $3000 per embryo, farmers would probably only be interested in very small numbers of cloned animals, most of which would be males. The relative importance of adult versus fetal cloning is discussed. The need for gene banks to preserve genetic variation is emphasized; both gametes and somatic tissue cultures should be considered.

  1. What's so bad about human cloning?

    PubMed

    Breitowitz, Yitzchok

    2002-12-01

    There appears to be a consensus in the general community that reproductive cloning is an immoral technology that should be banned. It may, however, be argued, at least from the perspective of the Jewish tradition, that reproductive cloning has many positive benefits. It is thus essential that one carefully weigh the costs and the benefits before deciding on a definitive course of action.

  2. Multiple phase estimation in quantum cloning machines

    NASA Astrophysics Data System (ADS)

    Yao, Yao; Ge, Li; Xiao, Xing; Wang, Xiao-guang; Sun, Chang-pu

    2014-08-01

    Since the initial discovery of the Wootters-Zurek no-cloning theorem, a wide variety of quantum cloning machines have been proposed aiming at imperfect but optimal cloning of quantum states within its own context. Remarkably, most previous studies have employed the Bures fidelity or the Hilbert-Schmidt norm as the figure of merit to characterize the quality of the corresponding cloning scenarios. However, in many situations, what we truly care about is the relevant information about certain parameters encoded in quantum states. In this work, we investigate the multiple phase estimation problem in the framework of quantum cloning machines, from the perspective of quantum Fisher information matrix (QFIM). Focusing on the generalized d-dimensional equatorial states, we obtain the analytical formulas of QFIM for both universal quantum cloning machine (UQCM) and phase-covariant quantum cloning machine (PQCM), and prove that PQCM indeed performs better than UQCM in terms of QFIM. We highlight that our method can be generalized to arbitrary cloning schemes where the fidelity between the single-copy input and output states is input-state independent. Furthermore, the attainability of the quantum Cramér-Rao bound is also explicitly discussed.

  3. Cloning of endangered mammalian species: any progress?

    PubMed

    Loi, Pasqualino; Galli, Cesare; Ptak, Grazyna

    2007-05-01

    Attempts through somatic cell nuclear transfer to expand wild populations that have shrunk to critical numbers is a logical extension of the successful cloning of mammals. However, although the first mammal was cloned 10 years ago, nuclear reprogramming remains phenomenological, with abnormal gene expression and epigenetic deregulation being associated with the cloning process. In addition, although cloning of wild animals using host oocytes from different species has been successful, little is known about the implication of partial or total mitochondrial DNA heteroplasmy in cloned embryos, fetuses and offspring. Finally, there is a need for suitable foster mothers for inter-intra specific cloned embryos. Considering these issues, the limited success achieved in cloning endangered animals is not surprising. However, optimism comes from the rapid gain in the understanding of the molecular clues underlying nuclear reprogramming. If it is possible to achieve a controlled reversal of the differentiated state of a cell then it is probable that other issues that impair the cloning of endangered animals, such as the inter-intra species oocyte or womb donor, will be overcome in the medium term.

  4. Hypoxically inducible barley lactate dehydrogenase: cDNA cloning and molecular analysis

    SciTech Connect

    Hondred, D. ); Hanson, A.D. Univ. de Montreal, Quebec )

    1990-09-01

    In the roots of barley and other cereals, hypoxia induces a set of five isozymes of L-lactate dehydrogenase (LDH; (S)-lactate:NADH oxidoreductase, EC 1.1.1.27). Biochemical and genetic data indicate that the five LDH isozymes are tetramers that arise from random association of the products of two Ldh loci. To investigate this system, cDNA clones of LDH were isolated from a {lambda}gt11 cDNA library derived from hypoxically treated barley roots. The library was screened with antiserum raised against barley LDH purified {approx}3,000-fold by an improved three-step procedure. Immunopositive clones were rescreened with a cDNA probe synthesized by the polymerase chain reaction using primers modeled from the amino acid sequences of two tryptic LDH peptides. Two types of LDH clones were found. Nucleotide sequence analysis of one representative insert of each type (respectively, 1,305 and 1,166 base pairs) revealed open reading framed encoding 10 peptide fragments of LDH. The 1,305-base-pair insert included the entire coding region of a 356-residue LDH monomer. The nucleotide sequences of the two LDH cDNAs were 92% identical in the coding region, but highly divergent in the 3{prime} noncoding region, and thus probably correspond to the two postulated Ldh loci. The deduced amino acid sequences of the two barley LDHs were 96% identical to each other and very similar to those from vertebrate and bacterial LDHs. RNA blot hybridization showed a single mRNA band of 1.5 kilobases whose level rose about 8-fold in roots during hypoxic induction, as did the level of translatable LDH message.

  5. "Goodbye Dolly?" The ethics of human cloning.

    PubMed Central

    Harris, J

    1997-01-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity. PMID:9451604

  6. In vivo cloning strategy for Rhizobium plasmids.

    PubMed

    Hernández-Lucas, I; Mavingui, P; Finan, T; Chain, P; Martínez-Romero, E

    2002-10-01

    We have developed a simple system to clone indigenous Rhizobium plasmids into E. coli. The strategy consists of three matings: the first is to insert Tn5 in the plasmid to be cloned, the second incorporates the integrative vector into the inserted Tn5 in the native Rhizobium plasmid, and the last mating transfers the target plasmid directly into E. coli. This mating-based system was successfully used to clone plasmids of Rhizobium species with sizes ranging from 150 to 270 kb. In addition, a 500-kb fragment of a 600-kb megaplasmid was also cloned. This strategy could be used for cloning indigenous replicons of other gram-negative bacteria into a different host.

  7. Chorioallantoic placenta defects in cloned mice

    SciTech Connect

    Wakisaka-Saito, Noriko; Kohda, Takashi . E-mail: tkhoda.epgn@tmd.ac.jp; Inoue, Kimiko; Ogonuki, Narumi; Miki, Hiromi; Hikichi, Takafusa; Mizutani, Eiji; Wakayama, Teruhiko; Kaneko-Ishino, Tomoko; Ogura, Atsuo; Ishino, Fumitoshi

    2006-10-13

    Somatic cell nuclear transfer technology has been applied to produce live clones successfully in several mammalian species, but the success rates are very low. In mice, about half of the nuclear transfer embryos undergo implantation, but very few survive to term. We undertook detailed histological analyses of placentas from cloned mouse embryos generated from cumulus cells at 10.5 dpc of pregnancy, by which stage most clones have terminated their development. At 10.5 dpc, the extraembryonic tissues displayed several defined histological patterns, each reflecting their stage of developmental arrest. The most notable abnormality was the poor development of the spongiotrophoblast layer of diploid cells. This is in contrast to the placental hyperplasia frequently observed in somatic clones at 12.5 dpc or later stages. A variety of structural abnormalities were also observed in the embryos. Both placental and embryonic defects likely contribute to the low success rate of the mouse clones.

  8. [Human clone or a delayed twin?].

    PubMed

    Szybalski, W

    2001-01-01

    Cloning is a natural mode of asexual reproduction for many organisms, which results in nearly identical copies of cells or organisms. In animals, including humans, identical twins are an example of natural cloning. In the case of sheep, scientists succeeded to produce the "delayed" identical twin. Dolly, of a mature animal by a rather complex and inefficient procedure. However, if this procedure is perfected, it will be useful to clone beloved pets and important laboratory animals. It will be much less suited for making (cloning) "delayed twin" of mature persons because of high costs together with present experimental uncertainties. The only required regulation for human cloning is that somebody must be legally, including financially, responsible for the results of such novel reproductive technique.

  9. "Goodbye Dolly?" The ethics of human cloning.

    PubMed

    Harris, J

    1997-12-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity.

  10. [Human cloning in Muslim and Arab law].

    PubMed

    Aldeeb Abu-Sahlieh, Sami A

    2009-01-01

    Cloning is a modern medical procedure that Muslim religious authorities treat en resorting to the general principles established by classical Muslim law based on the Koran and the Sunnah of Muhhamad as the messenger of God. In this regard, human beings are not capable of deciding what is or what is not lawful without resorting to divine norms. Cloning clashes with several principles. Firstly, the principle of the respect for life in relation to surpernumeraries, but Muslim authors are not in unanimous agreement on the determination of the moment at which life begins. Secondly, is the respect of progeny: cloning could only take place between a married couple. But even if these two principles are respected, cloning poses two major problems: the diversity of species expounded by the Koran and the Sunnah and a lack of interest. Which explains the quasi-unanimous opposition of Muslim writings regarding cloning.

  11. Meat and milk compositions of bovine clones.

    PubMed

    Tian, X Cindy; Kubota, Chikara; Sakashita, Kunihito; Izaike, Yoshiaki; Okano, Ryoichi; Tabara, Norio; Curchoe, Carol; Jacob, Lavina; Zhang, Yuqin; Smith, Sadie; Bormann, Charles; Xu, Jie; Sato, Masumi; Andrew, Sheila; Yang, Xiangzhong

    2005-05-03

    The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones.

  12. Meat and milk compositions of bovine clones

    PubMed Central

    Tian, X. Cindy; Kubota, Chikara; Sakashita, Kunihito; Izaike, Yoshiaki; Okano, Ryoichi; Tabara, Norio; Curchoe, Carol; Jacob, Lavina; Zhang, Yuqin; Smith, Sadie; Bormann, Charles; Xu, Jie; Sato, Masumi; Andrew, Sheila; Yang, Xiangzhong

    2005-01-01

    The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones. PMID:15829585

  13. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    PubMed Central

    Beyer, Hannes M.; Gonschorek, Patrick; Samodelov, Sophia L.; Meier, Matthias; Weber, Wilfried; Zurbriggen, Matias D.

    2015-01-01

    Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly), a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date. PMID:26360249

  14. Cleaning efficiency of nickel-titanium GT and .04 rotary files when used in a torque-controlled rotary handpiece.

    PubMed

    Suffridge, Calvin B; Hartwell, Gary R; Walker, Thomas L

    2003-05-01

    This study determined if the cleaning efficiency of nickel-titanium rotary files in an endodontic electric handpiece using a no-torque control setting was superior to that obtained when using the torque-control feature. Fifty extracted human anterior teeth with straight canals were divided into two groups of 20 and two control groups of 5. Canals were instrumented with GT and .04 ProFile nickel-titanium files until a size 35 advanced to working length. Samples were sectioned and the apical 6 mm of the canal was photographed (x20) and projected onto a 3- x 4-foot grid with squares measuring 0.5 inches each. Total debris was the percentage of the number of squares containing debris versus the total number of squares. The teeth in the torque-controlled group showed an average of 24.99% debris versus 15.55% for the teeth in the no-torque group. The difference was not statistically significant; therefore, no difference can be said to exist between the two torque settings in terms of cleaning efficiency.

  15. Study on Off-Design Steady State Performances of Helium Gas Turbo-compressor for HTGR-GT

    SciTech Connect

    Qisen Ren; Xiaoyong Yang; Zhiyong Huang; Jie Wang

    2006-07-01

    The high temperature gas-cooled reactor (HTGR) coupled with direct gas turbine cycle is a promising concept in the future of nuclear power development. Both helium gas turbine and compressor are key components in the cycle. Under normal conditions, the mode of power adjustment is to control total helium mass in the primary loop using gas storage vessels. Meanwhile, thermal power of reactor core is regulated. This article analyzes off-design performances of helium gas turbine and compressors for high temperature gas-cooled reactor with gas turbine cycle (HTGR-GT) at steady state level of electric power adjustment. Moreover, performances of the cycle were simply discussed. Results show that the expansion ratio of turbine decreases as electric power reduces but the compression ratios of compressors increase, efficiencies of both turbine and compressors decrease to some extent. Thermal power does not vary consistently with electric power, the difference between these two powers increases as electric power reduces. As a result of much thermal energy dissipated in the temperature modulator set at core inlet, thermal efficiency of the cycle has a widely reduction under partial load conditions. (authors)

  16. DIGLIB. PC-DOS Graphics Subroutine Library

    SciTech Connect

    Burleson, R.R.

    1989-02-01

    DIGLIB is a collection of general graphics subroutines. It was designed to be small, reasonably fast, device-independent, and compatible with DEC-supplied operating systems for VAXes, PDP-11s, and LSI-11s, and the DOS operating system for IBM PCs and IBM-compatible machines. DIGLIB/PC runs on IBM PCs under PC-DOS or MS-DOS. The software is readily usable by casual programmers for two-dimensional plotting.

  17. The golden anniversary of cloning: a celebratory essay.

    PubMed

    Di Berardino, Marie A; McKinnell, Robert G; Wolf, Don P

    2003-09-01

    May 2002 marked the golden anniversary of the first cloned tadpoles. We celebrate this anniversary, as nuclear transplantation of frog cells into enucleated eggs became the prototype for cloning insects, fish, and mammals. We briefly review the salient results from amphibian cloning. Extension of these studies to mammalian species led to cloning adult cells, important advances in understanding nuclear reprogramming, and the construction of transgenic clones for biomedical applications. In addition, murine cloning clarified two problems unresolved in frog cloning: the unequivocal demonstration that nuclei of fully differentiated cells can direct the formation of fertile adults, and that abnormal expression of genes was responsible for the endoderm and neural syndromes in Rana clones.

  18. Controlled secret sharing protocol using a quantum cloning circuit

    NASA Astrophysics Data System (ADS)

    Adhikari, Satyabrata; Roy, Sovik; Chakraborty, Shantanav; Jagadish, Vinayak; Haris, M. K.; Kumar, Atul

    2014-09-01

    We demonstrate the possibility of controlling the success probability of a secret sharing protocol using a quantum cloning circuit. The cloning circuit is used to clone the qubits containing the encoded information and en route to the intended recipients. The success probability of the protocol depends on the cloning parameters used to clone the qubits. We also establish a relation between the concurrence of initially prepared state, entanglement of the mixed state received by the receivers after cloning scheme and the cloning parameters of cloning machine.

  19. The POR rs1057868–rs2868177 GC-GT diplotype is associated with high tacrolimus concentrations in early post-renal transplant recipients

    PubMed Central

    Liu, Shu; Chen, Rong-xin; Li, Jun; Zhang, Yu; Wang, Xue-ding; Fu, Qian; Chen, Ling-yan; Liu, Xiao-man; Huang, Hong-bing; Huang, Min; Wang, Chang-xi; Li, Jia-li

    2016-01-01

    Aim: Cytochrome P450 oxidoreductase (POR) is the only flavoprotein that donates electrons to all microsomal P450 enzymes (CYP), and several POR SNPs have been shown to be important contributors to altered CYP activity or CYP-mediated drug metabolism. In this study we examined the association between 6 POR SNPs and tacrolimus concentrations in Chinese renal transplant recipients. Methods: A total of 154 renal transplant recipients were enrolled. Genotyping of CYP3A5*3 and 6 POR SNPs was performed. All patients received a triple immunosuppressive regimen comprising tacrolimus, mycophenolate mofetil and prednisone. Dose-adjusted tacrolimus trough concentrations were obtained on d 7 (C0D7/D) after transplantation when steady-state concentration of tacrolimus was achieved (dosage had been unchanged for more than 3 d). Results: Tacrolimus C0D7/D in CYP3A5*3/*3/ POR rs1057868–rs2868177 GC-GT diplotype carriers was 1.62- and 2.72-fold higher than those in CYP3A5*3/*3/ POR rs1057868–rs2868177 GC-GT diplotype non-carriers and CYP3A5*1 carriers (220.17±48.09 vs 135.69±6.86 and 80.84±5.27 ng/mL/mg/kg, respectively, P<0.0001). Of CYP3A5*3/*3/ POR rs1057868-rs2868177GC-GT diplotype carriers, 85.71% exceeded the upper limit of the target range (8 ng/mL), which was also significantly higher compared with the latter two groups (14.29% and 0.00%, respectively, P<0.0001). The CYP3A5*3 and POR rs1057868–rs2868177 GC-GT diplotype explained 31.7% and 5.7%, respectively, of the inter-individual variability of tacrolimus C0D7/D, whereas the POR rs1057868–rs2868177 GC-GT diplotype could explain 10.9% of the inter-individual variability of tacrolimus C0D7/D in CYP3A5 non-expressers. Conclusion: The CYP3A5*3 and POR rs1057868–rs2868177 GC-GT diplotype accounted for the inter-individual variation of tacrolimus C0D7/D. Genotyping of POR rs1057868–rs2868177 diplotypes would help to differentiate initial tacrolimus dose requirements and to achieve early target C0 ranges in Chinese

  20. The POR rs1057868-rs2868177 GC-GT diplotype is associated with high tacrolimus concentrations in early post-renal transplant recipients.

    PubMed

    Liu, Shu; Chen, Rong-Xin; Li, Jun; Zhang, Yu; Wang, Xue-Ding; Fu, Qian; Chen, Ling-Yan; Liu, Xiao-Man; Huang, Hong-Bing; Huang, Min; Wang, Chang-Xi; Li, Jia-Li

    2016-09-01

    Cytochrome P450 oxidoreductase (POR) is the only flavoprotein that donates electrons to all microsomal P450 enzymes (CYP), and several POR SNPs have been shown to be important contributors to altered CYP activity or CYP-mediated drug metabolism. In this study we examined the association between 6 POR SNPs and tacrolimus concentrations in Chinese renal transplant recipients. A total of 154 renal transplant recipients were enrolled. Genotyping of CYP3A5*3 and 6 POR SNPs was performed. All patients received a triple immunosuppressive regimen comprising tacrolimus, mycophenolate mofetil and prednisone. Dose-adjusted tacrolimus trough concentrations were obtained on d 7 (C0D7/D) after transplantation when steady-state concentration of tacrolimus was achieved (dosage had been unchanged for more than 3 d). Tacrolimus C0D7/D in CYP3A5*3/*3/ POR rs1057868-rs2868177 GC-GT diplotype carriers was 1.62- and 2.72-fold higher than those in CYP3A5*3/*3/ POR rs1057868-rs2868177 GC-GT diplotype non-carriers and CYP3A5*1 carriers (220.17±48.09 vs 135.69±6.86 and 80.84±5.27 ng/mL/mg/kg, respectively, P<0.0001). Of CYP3A5*3/*3/ POR rs1057868-rs2868177GC-GT diplotype carriers, 85.71% exceeded the upper limit of the target range (8 ng/mL), which was also significantly higher compared with the latter two groups (14.29% and 0.00%, respectively, P<0.0001). The CYP3A5*3 and POR rs1057868-rs2868177 GC-GT diplotype explained 31.7% and 5.7%, respectively, of the inter-individual variability of tacrolimus C0D7/D, whereas the POR rs1057868-rs2868177 GC-GT diplotype could explain 10.9% of the inter-individual variability of tacrolimus C0D7/D in CYP3A5 non-expressers. The CYP3A5*3 and POR rs1057868-rs2868177 GC-GT diplotype accounted for the inter-individual variation of tacrolimus C0D7/D. Genotyping of POR rs1057868-rs2868177 diplotypes would help to differentiate initial tacrolimus dose requirements and to achieve early target C0 ranges in Chinese renal transplant recipients.

  1. Tumor Necrosis Factor (TNF) –308G>A, Nitric Oxide Synthase 3 (NOS3) +894G>T Polymorphisms and Migraine Risk: A Meta-Analysis

    PubMed Central

    Chen, Min; Tang, Wenjing; Hou, Lei; Liu, Ruozhuo; Dong, Zhao; Han, Xun; Zhang, Xiaofei; Wan, Dongjun; Yu, Shengyuan

    2015-01-01

    Background and Objective Conflicting data have been reported on the association between tumor necrosis factor (TNF) –308G>A and nitric oxide synthase 3 (NOS3) +894G>T polymorphisms and migraine. We performed a meta-analysis of case-control studies to evaluate whether the TNF –308G>A and NOS3 +894G>T polymorphisms confer genetic susceptibility to migraine. Method We performed an updated meta-analysis for TNF –308G>A and a meta-analysis for NOS3 +894G>T based on studies published up to July 2014. We calculated study specific odds ratios (OR) and 95% confidence intervals (95% CI) assuming allele contrast, dominant model, recessive model, and co-dominant model as pooled effect estimates. Results Eleven studies in 6682 migraineurs and 22591 controls for TNF –308G>A and six studies in 1055 migraineurs and 877 controls for NOS3 +894G>T were included in the analysis. Neither indicated overall associations between gene polymorphisms and migraine risk. Subgroup analyses suggested that the “A” allele of the TNF –308G>A variant increases the risk of migraine among non-Caucasians (dominant model: pooled OR = 1.82; 95% CI 1.15 – 2.87). The risk of migraine with aura (MA) was increased among both Caucasians and non-Caucasians. Subgroup analyses suggested that the “T” allele of the NOS3 +894G>T variant increases the risk of migraine among non-Caucasians (co-dominant model: pooled OR = 2.10; 95% CI 1.14 – 3.88). Conclusions Our findings appear to support the hypothesis that the TNF –308G>A polymorphism may act as a genetic susceptibility factor for migraine among non-Caucasians and that the NOS3 +894G>T polymorphism may modulate the risk of migraine among non-Caucasians. PMID:26098763

  2. Tracking the Evolution of Code Clones

    NASA Astrophysics Data System (ADS)

    Bakota, Tibor

    It is believed by many academic and industrial experts, that source code cloning (copy&paste programming) represents a significant threat to maintainability in an evolving software system. The real threat does not lie in the existence of duplications, but the fears are in connection with their evolution. There exist an abundance of algorithms for finding code clones in one particular version of a software system, but eliminating or even evaluating these clones often seems hopeless, as there may exist several thousands of them. Tracking the evolution of individual clones might solve the problem, as it could help identifying the inconsistently changing duplications: the clones which are really dangerous at a particular moment. In this paper we present an approach for mapping code duplications from one particular version of the software to another one, based on a similarity distance function. For the suspicious evolution patterns we introduce the term of "clone smells". By defining the relevant categories of the possible evolution patterns, the proposed method also gives a clue about why the reported code fragments might be dangerous. In the case study, clone smells were extracted, evaluated, and manually categorized throughout many versions of the jEdit system. The findings suggest that roughly half of the reported smells refer to inconsistent changes in the code.

  3. Human embryo cloning prohibited in Hong Kong.

    PubMed

    Liu, Athena

    2005-12-01

    Since the birth of Dolly (the cloned sheep) in 1997, debates have arisen on the ethical and legal questions of cloning-for-biomedical-research (more commonly termed "therapeutic cloning") and of reproductive cloning using human gametes. Hong Kong enacted the Human Reproductive Technology Ordinance (Cap 561) in 2000. Section 15(1)(e) of this Ordinance prohibits the "replacing of the nucleus of a cell of an embryo with a nucleus taken from any other cell," i.e., nucleus substitution. Section 15(1)(f) prohibits the cloning of any embryo. The scope of the latter, therefore, is arguably the widest, prohibiting all cloning techniques such as cell nucleus replacement, embryo splitting, parthenogenesis, and cloning using stem cell lines. Although the Human Reproductive Technology Ordinance is not yet fully operative, this article examines how these prohibitions may adversely impact on basic research and the vision of the Hong Kong scientific community. It concludes that in light of recent scientific developments, it is time to review if the law offers a coherent set of policies in this area.

  4. Who is the parent in cloning?

    PubMed

    Elster, N

    1999-01-01

    In July 1996, a sheep named Dolly was born in Scotland. What makes Dolly's birth noteworthy is that she is the result of the first successful cloning attempt using the nucleus of an adult cell. The technique that led to Dolly's birth involved transferring the nucleus of a mammary cell from an adult sheep to the enucleated egg cell of an unrelated sheep with gestation occurring in a third sheep. The possibility of applying this technique to human reproduction raised concerns worldwide with several countries moving for an immediate bans on human cloning. In the United States, President Clinton requested that the National Bioethics Advisory Commission ("NBAC"), a multidisciplinary group composed of scientists, lawyers, educators, theologians, and ethicists study the implications of cloning and issue recommendations. The Commission consulted other scientists, ethicists, theologians, lawyers, and citizens with interests in this advancing technology and concluded that, "at this time it is morally unacceptable for anyone in the public or private sector, whether in a research or clinical setting, to attempt to create a child using somatic cell nuclear transfer cloning." This Article was included in a larger work prepared at the request of, and submitted to the Commission by, law professor Lori B. Andrews. Cloning through nuclear transfer will change the way we create and define families. This Article explores how existing law relating to parentage, surrogacy, egg donation, and artificial insemination may apply in the cloning context to clarify the parent-child relationship established through cloning.

  5. [Worldviews and philosophical basis of human cloning].

    PubMed

    Lukowska, A T

    2001-01-01

    The article presents three standpoints on the question of moral permissibility of human cloning and shows the philosophical principles of it. 1. The moral consent to human cloning with the purposes of reproduction and therapy. The followers of human cloning refer to materialistic anthropology also to subjectivistic, relativistic and utilitarian ethics. 2. Those, who are adverse to human cloning with the purpose of reproduction, but they acquiesce to the so-called therapeutic cloning. They reject that human embryos and foetuses are individuals who come under protection of law. 3. Those, who reject human cloning for the purposes of reproduction and therapy alike. They assent to a personalistic anthropology and Christian ethics. A human being was created by God and human life begins at the moment of insemination. All three groups use various argumentation. The arguments for and against cloning are extracted from biology as well as psychology, philosophy, law and religion. The author of the article takes the last standpoint, but she does not see such arguments, that might convince the opposite parties to a suit.

  6. Economical quantum cloning in any dimension

    SciTech Connect

    Durt, Thomas; Fiurasek, Jaromir; Cerf, Nicolas J.

    2005-11-15

    The possibility of cloning a d-dimensional quantum system without an ancilla is explored, extending on the economical phase-covariant cloning machine for qubits found in Phys. Rev. A 60, 2764 (1999). We prove the impossibility of constructing an economical version of the optimal universal 1{yields}2 cloning machine in any dimension. We also show, using an ansatz on the generic form of cloning machines, that the d-dimensional 1{yields}2 phase-covariant cloner, which optimally clones all balanced superpositions with arbitrary phases, can be realized economically only in dimension d=2. The used ansatz is supported by numerical evidence up to d=7. An economical phase-covariant cloner can nevertheless be constructed for d>2, albeit with a slightly lower fidelity than that of the optimal cloner requiring an ancilla. Finally, using again an ansatz on cloning machines, we show that an economical version of the 1{yields}2 Fourier-covariant cloner, which optimally clones the computational basis and its Fourier transform, is also possible only in dimension d=2.

  7. Serial cloning of pigs by somatic cell nuclear transfer: restoration of phenotypic normality during serial cloning.

    PubMed

    Cho, Seong-Keun; Kim, Jae-Hwan; Park, Jong-Yi; Choi, Yun-Jung; Bang, Jae-Il; Hwang, Kyu-Chan; Cho, Eun-Jeong; Sohn, Sea-Hwan; Uhm, Sang Jun; Koo, Deog-Bon; Lee, Kyung-Kwang; Kim, Teoan; Kim, Jin-Hoi

    2007-12-01

    Somatic cell nuclear transfer (scNT) is a useful way to create cloned animals. However, scNT clones exhibit high levels of phenotypic instability. This instability may be due to epigenetic reprogramming and/or genomic damage in the donor cells. To test this, we produced transgenic pig fibroblasts harboring the truncated human thrombopoietin (hTPO) gene and used them as donor cells in scNT to produce first-generation (G1) cloned piglets. In this study, 2,818 scNT embryos were transferred to 11 recipients and five G1 piglets were obtained. Among them, a clone had a dimorphic facial appearance with severe hypertelorism and a broad prominent nasal bridge. The other clones looked normal. Second-generation (G2) scNT piglets were then produced using ear cells from a G1 piglet that had an abnormal nose phenotype. We reasoned that, if the phenotypic abnormality of the G1 clone was not present in the G2 and third-generation (G3) clones, or was absent in the G2 clones but reappeared in the G3 clones, the phenotypic instability of the G1 clone could be attributed to faulty epigenetic reprogramming rather than to inherent/accidental genomic damage to the donor cells. Blastocyst rates, cell numbers in blastocyst, pregnancy rates, term placenta weight and ponderal index, and birth weight between G1 and G2 clones did not differ, but were significantly (P < 0.05) lower than control age- and sex-matched piglets. Next, we analyzed global methylation changes during development of the preimplantation embryos reconstructed by donor cells used for the production of G1 and G2 clones and could not find any significant differences in the methylation patterns between G1 and G2 clones. Indeed, we failed to detect the phenotypic abnormality in the G2 and G3 clones. Thus, the phenotypic abnormality of the G1 clone is likely to be due to epigenetic dysregulation. Additional observations then suggested that expression of the hTPO gene in the transgenic clones did not appear to be the cause of the

  8. Molecular cloning and characterization of interferon. alpha. /. beta. response element binding factors of the murine (2 prime -5 prime )oligoadenylate synthetase ME-12 gene

    SciTech Connect

    Yan, Cong; Tamm, Igor )

    1991-01-01

    Seven clones encoding interferon response element binding factors have been isolated from a mouse fibroblast {lambda}gt11 cDNA library by using a {sup 32}P end-labeled tandem trimer of the mouse (2{prime}-5{prime})oligoadenylate synthetase gene interferon response element as a probe. Clone 16 shares strong similarity (95%) at both DNA and amino acid level with YB-1, a human major histocompatibility complex class II Y-box DNA-binding protein, and with dbpB, a human epidermal growth factor receptor gene enhancer region binding protein. The product of the gene represented by clone 16 may represent a factor that regulates multiple genes by binding to a variety of 5{prime} regulatory elements. Clone 25 is a 2407-base-pair-long cDNA and contains a putative 311-amino acid open reading frame corresponding to an estimated mass of 35.5 kDa. This putative protein, designated as interferon resonse element binding factor 1 (IREBF-1), contains an acidic domain, three heptad repeat leucine arrays, and a region that shares similarity with the yeast transcriptional factor CAL4 DNA-binding domain. Furthermore, the C terminus of IREBF-1 shows an unusual amphipathic property: within a 79-amino acid range, one side of the {alpha}-helical region contains a preponderance of hydrophobic amino acids and the other side contains hydrophilic amino acids. This type of structure provides a strong hydrophobic force for protein-protein interaction.

  9. Phase-covariant cloning of coherent states

    NASA Astrophysics Data System (ADS)

    Sacchi, Massimiliano F.

    2007-04-01

    We consider the problem of phase-covariant cloning for coherent states. We show that an experimental scheme based on ideal phase measurement and feedforward outperforms the semiclassical procedure of ideal phase measurement and preparation in terms of fidelity. A realistic scheme where the ideal phase measurement is replaced with double-homodyne detection is shown to be unable to overcome the semiclassical cloning strategy. On the other hand, such a realistic scheme is better than semiclassical cloning based on double-homodyne phase measurement and preparation.

  10. [Cloning: applications in humans 1. Technical aspects].

    PubMed

    Geraedts, J P; de Wert, G M

    2001-04-01

    The successful cloning experiments in mammals such as the sheep and mouse prompted speculations on clinical application in humans. Cloning is possible by nucleus transplantation and by embryo splitting. Nucleus transplantation does not result in a genetically completely identical individual because the mitochondrial DNA originates from the ovum donor. Embryo splitting may be regarded as the artificial production of a monozygotic multiplet. Possible applications of cloning in humans belong in the context of reproduction (treatment of couples with subfertility, with genetic problems or with a 'replica motive'), transplantation of genetically identical tissue, and scientific research.

  11. [Cloning: applications in humans. I. Technical aspects].

    PubMed

    Geraedts, J P; de Wert, G M

    2000-05-13

    The successful cloning experiments in mammals such as the sheep and mouse prompted speculations on clinical application in humans. Cloning is possible by nucleus transplantation and by embryo splitting. Nucleus transplantation does not result in a genetically completely identical individual because the mitochondrial DNA originates from the ovum donor. Embryo splitting may be regarded as the artificial production of a monozygotic multiplet. Possible applications of cloning in humans belong in the context of reproduction (treatment of couples with subfertility, with genetic problems or with a 'replica motive'), transplantation of genetically identical tissue, and scientific research.

  12. GT-12 - EARTH SKY

    NASA Image and Video Library

    1966-11-14

    S66-63030 (14 Nov. 1966) --- Gulf Coast area from Matagorda Bay, Texas, to Vermillion Bay, Louisiana, looking east, as seen from the Gemini-12 spacecraft during its 44th revolution of Earth. Galveston Bay is in center of picture. Houston and its environs are clearly visible. Note network of freeways and superhighways. Large lake near left center of picture is the Sam Rayburn Reservoir. Photo credit: NASA

  13. GT-7 RECOVERY

    NASA Image and Video Library

    1965-12-18

    S65-61830 (18 Dec. 1965) --- Astronauts James A. Lovell Jr. (left), Gemini-7 pilot, and Frank Borman, command pilot, are shown just after they arrived aboard the aircraft carrier USS Wasp. Greeting the astronauts are Donald Stullken (at Lovell's right), Recovery Operations Branch, Landing and Recovery Division, Dr. Howard Minners (standing beside Borman), Flight Medicine Branch, Center Medical Office, Manned Spacecraft Center, and Bennett James (standing behind Borman), a NASA Public Affairs Officer. The National Aeronautics and Space Administration's Gemini-7 spacecraft splashed down in the western Atlantic recovery area at 9:05 a.m. (EST), Dec. 18, 1965, to conclude a record-breaking 14-day mission in space. Photo credit: NASA

  14. GT-7 RECOVERY

    NASA Image and Video Library

    1965-12-18

    S65-63690 (18 Dec. 1965) --- Astronauts Frank Borman, command pilot, and James A. Lovell Jr., pilot, sit in life raft while awaiting pickup by a helicopter from the aircraft carrier USS Wasp. The three-man Navy frogman team attached the flotation collar to increase the Gemini-7 spacecraft's buoyancy prior to recovery. Photo credit: NASA

  15. GT-12 - RECOVERY

    NASA Image and Video Library

    1966-11-15

    S66-59936 (15 Nov. 1966) --- The last Gemini spaceflight is concluded as the Gemini-12 spacecraft, with astronaut James A. Lovell Jr., command pilot, and Edwin E. Aldrin Jr., pilot, aboard, touches down in the Atlantic Ocean 2.5 nautical miles from the prime recovery ship, USS Wasp. Gemini-12 splashed down at 2:21 p.m. (EST), Nov. 15, 1966, to conclude a four-day mission in space. Photo credit: NASA

  16. GT-7 recovery

    NASA Image and Video Library

    1965-12-18

    S65-63644 (18 Dec. 1965) --- Crewmen of the aircraft carrier USS Wasp gather on deck to watch the recovery of the National Aeronautics and Space Administration's Gemini-7 spacecraft and astronauts. Gemini-7, with astronauts Frank Borman, command pilot, and James A. Lovell Jr., pilot, splashed down in the western Atlantic at 9:05 a.m. (EST), Dec. 18, 1965, to conclude a record-breaking 14-day mission in space. Photo credit: NASA

  17. Debris - Onboard GT-10

    NASA Image and Video Library

    1966-07-18

    S66-46241 (18 July 1966) --- Debris on spacecraft window in picture taken from inside the Gemini-10 spacecraft. At this time Gemini-10 was docked with the Agena Target Docking Vehicle 5005. Taken with a modified 70mm Hasselblad camera, using Eastman Kodak, Ektachrome, MS (S.O. 217) color film. The Gemini-10 crew is astronaut John W. Young, command pilot, and Michael Collins, pilot. Photo credit: NASA

  18. PREFLIGHT MEDICAL (GT-7)

    NASA Image and Video Library

    1965-12-02

    S65-56315 (2 Dec. 1965) --- Dr. Charles A. Berry (left), chief of the Manned Spacecraft Center (MSC) Medical Programs, and astronauts James A. Lovell Jr. (center), Gemini-7 pilot, and Frank Borman, Gemini-7 command pilot, examine a series of chest x-rays taken during the preflight physical. Photo credit: NASA

  19. Earth Sky GT-7

    NASA Image and Video Library

    1965-12-07

    S65-63829 (5 Dec. 1965) --- Algeria, south of the Fort Flatters area, as seen from the National Aeronautics and Space Administration?s Gemini-7 spacecraft during its 13th revolution of Earth. The orange color area is the Tifermine Sand Dunes that reach a height of 1,000 feet. The photograph was taken with a modified 70mm Hasselblad camera, with Eastman Kodak, Ektachrome MS (S.O. 217) color film. Photo credit: NASA

  20. Earth Sky GT-7

    NASA Image and Video Library

    1965-12-07

    S65-63784 (7 Dec. 1965) --- Algeria, south of Celemb Bechar, as seen from the orbiting Gemini-7 spacecraft during its 42nd revolution of Earth. Note rain runoff on the desert floor. Astronaut Frank Borman and James A. Lovell Jr. were aboard the National Aeronautics and Space Administration?s Gemini-7 spacecraft. The photograph was taken with a modified 70mm Hasselblad camera, using Eastman Kodak, Ektachrome MS (S.O. 217) color film. Photo credit: NASA

  1. MCC During GT-11

    NASA Image and Video Library

    1966-09-15

    S66-52762 (15 Sept. 1966) --- Dr. Robert R. Gilruth (left) smokes a cigar in Houston's Mission Control Center to celebrate the successful splashdown of Gemini-11. Looking on are James C. Elms (center), MSC deputy director, and Charles W. Mathews, Gemini program manager. Photo credit: NASA

  2. GT-9 TEST - TRAINING

    NASA Image and Video Library

    1965-04-14

    S65-22639 (14 April 1965) --- The Gemini-Titan 4 prime crew, astronauts Edward H. White II (left), pilot; and James A. McDivitt (getting in the spacecraft), command pilot, received instructions from Gordon Harvey, Flight Crew Support Division; and Alan M. Rochford, suit technician, Crew Systems Division, before closing of hatches prior to undergoing water egress training in the Gulf of Mexico.

  3. Earth Sky- GT-7

    NASA Image and Video Library

    1965-12-12

    S65-63766 (12 Dec. 1965) --- Ras Azir on the coast of the Republic of Somali, looking east, as seen from the National Aeronautics and Space Administration's Gemini-7 spacecraft during its 117th revolution of Earth. Taken with a modified 70mm Hasselblad camera, using Eastman Kodak, Ektachrome MS (S.O. 217) color film. Photo credit: NASA

  4. Cloning of rat homeobox genes

    SciTech Connect

    Sakoyama, Yasuhiko; Mizuta, Ikuko; Ogasawara, Naotake

    1994-10-01

    We report the isolation of nine rat cognates of mouse homeoboxes within the four Hox gene clusters and a rat homologue of mouse IPF1 homeobox, RHbox No. 13A. The sequences of nine cloned homeoboxes are highly similar to those of the mouse and human homeoboxes in the Hox clusters. The restriction enzyme sites and map distances between each of the homeoboxes on the rat genome are nearly identical to those of mouse and human. Thus, we conclude that the isolated homeoboxes are the rat homologues of mouse homeoboxes within the four Hox clusters. A novel homeobox RHbox No. 13A is different from the Drosophila Antennapedia (Antp) sequence but is highly similar to the XlHbox8 (Xenopus laevis) and HtrA2 (Helobdella triserialis) homeoboxes. Forty-two amino acids of the last two-thirds of the RHbox No. 13A, XlHbox8, and mouse IPF1 homeodomains completely matched. In addition, these four homeodomains contain a unique His residue in the recognition helix of a helix-turn-helix DNA-binding motif. This His residue is not found in any of the previously published mammalian homeodomain sequences except mouse IPF1. 24 refs., 4 figs.

  5. [Comparative embryology and mammalian cloning].

    PubMed

    Sakharova, N Iu; Chaĭlakhian, L M

    2010-01-01

    A hypothesis has been advanced that logically combines "contradictory" facts concerning the early mammalian development and shows a natural relationship between the embryos developing from a fertilized ovum and from cells of the inner cell mass of blastocyst. When studying the theoretical questions of cloning, it is necessary to take into consideration the peculiarities of prenatal mammalian ontogenesis, which make themselves evident upon comparison with other animals. The absence of yolk in the mammalian ovum defines sharp differences in the early development between mammals and other Amniota. The whole asynchronic cleavage results in the formation of the morula followed by the blastocyst, which hatches from zona pellucida and is implanted into the uterus tissue. This fact allows us to consider the blastocyst as a mammalian larva, which is fed thanks to maternal organism. It is known that, in the body of a larva (blastocyst), a new embryo develops from some somatic cells. This process is known as a polyembryony, which is typical for the development of some parasitic insects. The polyembryony in turn is a variant of somatic embryogenesis, which is a form of asexual reproduction. Thus, two different embryos, "conceptus" and "embryo proper", have different origin: the first forms by the sexual way and the second, by the asexual. The investigation of the mechanisms of somatic embryogenesis in mammals will help us to find conditions necessary for the full reprograming of donor somatic nuclei and provide the successful development of reconstructed embryos.

  6. Differential protein expression profile in the hypothalamic GT1-7 cell line after exposure to anabolic androgenic steroids

    PubMed Central

    Martínez-Rivera, Freddyson J.; Pérez-Laspiur, Juliana; Santiago-Gascot, María E.; Alemán-Reyes, Abner G.; García-Santiago, Emanuel; Rodríguez-Pérez, Yolanda; Calo-Guadalupe, Cristhian; Otero-Pagán, Inelia; Ayala-Pagán, Roxsana N.; Martínez, Magdiel; Cantres-Rosario, Yisel M.; Meléndez, Loyda M.; Barreto-Estrada, Jennifer L.

    2017-01-01

    The abuse of anabolic androgenic steroids (AAS) has been considered a major public health problem during decades. Supraphysiological doses of AAS may lead to a variety of neuroendocrine problems. Precisely, the hypothalamic-pituitary-gonadal (HPG) axis is one of the body systems that is mainly influenced by steroidal hormones. Fluctuations of the hormonal milieu result in alterations of reproductive function, which are made through changes in hypothalamic neurons expressing gonadotropin-releasing hormone (GnRH). In fact, previous studies have shown that AAS modulate the activity of these neurons through steroid-sensitive afferents. To increase knowledge about the cellular mechanisms induced by AAS in GnRH neurons, we performed proteomic analyses of the murine hypothalamic GT1-7 cell line after exposure to 17α-methyltestosterone (17α-meT; 1 μM). These cells represent a good model for studying regulatory processes because they exhibit the typical characteristics of GnRH neurons, and respond to compounds that modulate GnRH in vivo. Two-dimensional difference in gel electrophoresis (2D-DIGE) and mass spectrometry analyses identified a total of 17 different proteins that were significantly affected by supraphysiological levels of AAS. Furthermore, pathway analyses showed that modulated proteins were mainly associated to glucose metabolism, drug detoxification, stress response and cell cycle. Validation of many of these proteins, such as GSTM1, ERH, GAPDH, PEBP1 and PDIA6, were confirmed by western blotting. We further demonstrated that AAS exposure decreased expression of estrogen receptors and GnRH, while two important signaling pathway proteins p-ERK, and p-p38, were modulated. Our results suggest that steroids have the capacity to directly affect the neuroendocrine system by modulating key cellular processes for the control of reproductive function. PMID:28719635

  7. Hydrogen Fueled Hybrid Solid Oxide Fuel Cell-Gas Turbine (SOFC-GT) System for Long-Haul Rail Application

    NASA Astrophysics Data System (ADS)

    Chow, Justin Jeff

    Freight movement of goods is the artery for America's economic health. Long-haul rail is the premier mode of transport on a ton-mile basis. Concerns regarding greenhouse gas and criteria pollutant emissions, however, have motivated the creation of annually increasing locomotive emissions standards. Health issues from diesel particulate matter, especially near rail yards, have also been on the rise. These factors and the potential to raise conventional diesel-electric locomotive performance warrants the investigation of using future fuels in a more efficient system for locomotive application. This research evaluates the dynamic performance of a Solid Oxide Fuel Cell-Gas Turbine (SOFC-GT) Hybrid system operating on hydrogen fuel to power a locomotive over a rail path starting from the Port of Los Angeles and ending in the City of Barstow. Physical constraints, representative locomotive operation logic, and basic design are used from a previous feasibility study and simulations are performed in the MATLAB Simulink environment. In-house controls are adapted to and expanded upon. Results indicate high fuel-to-electricity efficiencies of at least 54% compared to a conventional diesel-electric locomotive efficiency of 35%. Incorporation of properly calibrated feedback and feed-forward controls enables substantial load following of difficult transients that result from train kinematics while maintaining turbomachinery operating requirements and suppressing thermal stresses in the fuel cell stack. The power split between the SOFC and gas turbine is deduced to be a deterministic factor in the balance between capital and operational costs. Using hydrogen results in no emissions if renewable and offers a potential of 24.2% fuel energy savings for the rail industry.

  8. Cholesterol secoaldehyde, an ozonation product of cholesterol, induces amyloid aggregation and apoptosis in murine GT1-7 hypothalamic neurons.

    PubMed

    Sathishkumar, K; Xi, Xiaochun; Martin, Roy; Uppu, Rao M

    2007-06-01

    Aldehydic products from ozonation of cholesterol and peroxidation of phospholipids have been shown to accelerate aggregation of amyloid-beta (Abeta) in vitro. Here, we show that 3beta-hydroxy-5-oxo-5,6-secocholestan-6-al (ChSeco), an ozonation product of cholesterol, induces Abeta aggregation, generation of reactive oxygen species (ROS), and cytotoxicity in murine GT1-7 hypothalamic neurons. The formation of Abeta aggregates in situ was dose-dependent at ChSeco concentrations ranging from 1 to 20 microM. The increase in insoluble Abeta aggregates at increasing concentrations of ChSeco was accompanied by a decrease in soluble Abeta as evidenced by Western blot analysis. The formation of ROS in neuronal cells was found to be dose- and time-dependent with the magnitude being higher at 20 microM compared to 10 microM ChSeco or untreated controls. The increase in ROS was associated with depletion of GSH. The cytotoxicity induced by ChSeco involved changes in phosphatidylserine translocation, DNA fragmentation, and caspase 3/7 activity that are characteristic of apoptosis. Pretreatment of neuronal cells with Trolox, a water-soluble analog of alpha-tocopherol offered partial, but significant protection against ChSeco-induced cell death, whereas, N-acetyl-L-cysteine (NAC) completely prevented the cytotoxic effects of ChSeco. NAC and Trolox were without any effects on ChSeco-induced Abeta aggregation. Fibrillogenesis inhibitors, which inhibited Abeta aggregation, did not inhibit cell death induced by ChSeco, implying that ROS generation, and not Abeta aggregation, plays a major role in the observed cytotoxicity. However, since Alzheimer's and other neurodegenerative diseases are slow and progressive, the formation of Abeta aggregates in vivo by ChSeco may have long-term pathological consequences.

  9. Association of the DNMT3B -579G>T Polymorphism with Risk of Thymomas in Patients with Myasthenia Gravis

    PubMed Central

    Coppedè, Fabio; Ricciardi, Roberta; Denaro, Maria; De Rosa, Anna; Provenzano, Carlo; Bartoccioni, Emanuela; Baggiani, Angelo; Lucchi, Marco; Mussi, Alfredo; Migliore, Lucia

    2013-01-01

    Increasing evidence suggests a contribution of epigenetic processes in promoting cancer and autoimmunity. Myasthenia gravis (MG) is an autoimmune disease mediated, in approximately 80% of the patients, by antibodies against the nicotinic acetylcholine receptor (AChR+). Moreover, epithelial tumours (thymomas) are present in about 10-20% of the patients, and there is indication that changes in DNA methylation might contribute to the risk and progression of thymomas. However, the role of epigenetics in MG is still not completely clarified. In the present study we investigated if a common polymorphism (-579G>T: rs1569686) in the promoter of the DNMT3B gene coding for the DNA methyltransferase 3B, an enzyme that mediates DNA methylation, increases the risk to develop MG or MG-associated thymomas. The study polymorphism was selected based on recent reports and a literature meta-analysis suggesting association with increased risk of various types of cancer. We screened 324 AChR+ MG patients (140 males and 184 females, mean age 56.0 ± 16.5 years) and 735 healthy matched controls (294 males and 441 females, mean age 57.3 ± 15.6 years). 94 of the total MG patients had a thymoma. While there was no association with the whole cohort of MG patients, we found a statistically significant association of the DNMT3B -579T allele (OR = 1.51; 95% CI=1.1-2.1, P = 0.01) and the TT homozygous genotype (OR = 2.59; 95% CI=1.4-4.9, P = 0.006) with the risk of thymoma. No association was observed in MG patients without thymoma, even after stratification into clinical subtypes. Present results suggest that the DNMT3B -579T allele might contribute to the risk of developing thymoma in MG patients, particularly in homozygous TT subjects. PMID:24260492

  10. Glutamate increases toxicity of inorganic lead in GT1-7 neurons: partial protection induced by flunarizine.

    PubMed

    Loikkanen, Jarkko; Naarala, Jonne; Vähäkangas, Kirsi H; Savolainen, Kai M

    2003-12-01

    Recent studies point to an interaction between the glutamatergic neurotransmitter system and inorganic lead (Pb) neurotoxicity. Pb (1-100 microM) evoked cytotoxicity over the period of 72 h in mouse hypothalamic GT1-7 neurons. Glutamate (0.1 or 1 mM) on its own did not have any effect on cell viability. However, 1 mM glutamate clearly increased Pb-induced cell death at 48 and 72 h. Although flunarizine (0.1-10 microM), an antagonist of L- and T-type voltage-sensitive calcium channels (VSCCs), partially protected from the cytotoxicity induced by co-exposure to Pb (10 or 100 micro M) and glutamate (1 mM), it had no protective effect on cytotoxicity induced by Pb alone. The flunarizine-induced protection was dependent on time and observed only at 48 h. Neither verapamil, an antagonist of L-type VSCCs, nor DIDS, an inhibitor of anion exchange, at non-toxic concentrations (0.1-10 microM) had any effect on cytotoxicity induced by Pb alone or together with glutamate at any studied time point. Co-exposure to Pb and glutamate also resulted in more prominent production of reactive oxygen species (ROS) than either of the compounds alone. Interestingly, we observed an increase in intracellular glutathione (GSH) levels in cells exposed to micromolar concentrations of Pb. Glutamate decreased the levels of intracellular GSH and also partially reduced the Pb-induced increase in GSH levels. These results suggest that the interaction of glutamate and Pb results in increased neuronal cell death via mechanisms that involve an increase in ROS production, a decrease in intracellular GSH defense against oxidative stress and probably T-type VSCCs.

  11. Comparing the acceptor promiscuity of a Rosa hybrida glucosyltransferase RhGT1 and an engineered microbial glucosyltransferase OleD(PSA) toward a small flavonoid library.

    PubMed

    Wang, Lu; Han, Weiqing; Xie, Chenying; Hou, Jingli; Fang, Qinghong; Gu, Jianchun; Wang, Peng George; Cheng, Jiansong

    2013-03-07

    Glycosylation is a widespread modification of plant secondary metabolites, and catalyzed by a superfamily of enzymes called UDP-glycosyltransferases (UGTs). UGTs are often involved in late biosynthetic steps and show broad substrate specificity or regioselectivity. In this study, the acceptor promiscuity of a Rosa hybrid UGT RhGT1 and an evolved microbial UGT OleD(PSA) toward a small flavonoid library was probed and compared. Interestingly, RhGT1 showed comparable acceptor promiscuity in comparison with OleD(PSA), though the acceptor binding pocket of the latter is much more open and large. This clearly indicates that stabilization of the acceptor position by suitable hydrophobic interactions is important for the specificity or regioselectivity determination as well as overall fit of the acceptor into a 'big enough' binding pocket. This also poses a challenge for structure-based UGT engineering to alter the glucosylation pattern of flavonoids. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Association of RAD 51 135 G/C, 172 G/T and XRCC3 Thr241Met gene polymorphisms with increased risk of head and neck cancer.

    PubMed

    Kayani, Mahmood Akhtar; Khan, Sumeera; Baig, Ruqia Mehmood; Mahjabeen, Ishrat

    2014-01-01

    Homologous recombination repair (HRR) plays an important role in protection against carcinogenic factors. Genes regulating the HRR mechanisms may impair their functions and consequently result in increased cancer susceptibility. RAD 51 and XRCC3 are key regulators of the HRR pathway and genetic variability in these may contribute to the appearance and progression of various cancers including head and neck cancer (HNC). The aim of the present study was to compare the distribution of genotypes of RAD51 (135G/C, 172 G/T) and XRCC3 (Thr241Met) polymorphisms between HNC patients and controls. Each polymorphism was genotyped using the polymerase chain reaction-restriction fragment length polymerase (PCR-RFLP) technique in 200 pathologically confirmed HNC patients along with 150 blood samples from normal, disease free healthy individuals. We observed that homozygous variant CC genotype of RAD51 135G/C was associated with a 2.5 fold increased HNC risk (OR=2.5; 95%CI=0.69-9.53; p<0.02), while second polymorphism of RAD 51 172 G/T, heterozygous variant GT genotype was associated with a 1.68 fold (OR=1.68; 95%CI=1.08-2.61; p<0.02) elevation when compared with controls. In the case of the Thr241Met polymorphism of XRCC3, we observed a 16 fold (OR=16; 95% CI= 3.78-69.67; p<0.0002) increased HNC risk in patients compared to controls. These results further suggested that RAD51 (135G/C, 172 G/T) and XRCC3 (Thr241Met) polymorphisms may be effective biomarkers for genetic susceptibility to HNC. Larger studies are needed to confirm our findings and identify the underlying mechanisms.

  13. Generation of phase-covariant quantum cloning

    NASA Astrophysics Data System (ADS)

    Karimipour, V.; Rezakhani, A. T.

    2002-11-01

    It is known that in phase-covariant quantum cloning, the equatorial states on the Bloch sphere can be cloned with a fidelity higher than the optimal bound established for universal quantum cloning. We generalize this concept to include other states on the Bloch sphere with a definite z component of spin. It is shown that once we know the z component, we can always clone a state with a fidelity higher than the universal value and that of equatorial states. We also make a detailed study of the entanglement properties of the output copies and show that the equatorial states are the only states that give rise to a separable density matrix for the outputs.

  14. [Therapeutic cloning. Biology, perspectives and alternatives].

    PubMed

    Maddox-Hyttel, Poul

    2003-02-24

    Certain diseases are caused by or cause irreversible loss of cells and may in the future be treated by cell-based therapies where spare cells are introduced into the body. Therapeutic cloning constitutes a scientifically and ethically challenging route to the generation of autologous patient specific spare cells: Stem cells for subsequent differentiation and transplantation are isolated from one week old embryos, which are produced by cloning by nuclear transfer from normal cells retrieved from a patient. Research in therapeutic cloning should be pursued in line with alternative strategies for obtaining stem cells. Finally, the molecular biology of cloning by nuclear transfer may hold the key to understanding trans-differentiation, which ultimately may allow for de-differentiation and subsequent re-differentiation of adult somatic cells for therapeutic purposes.

  15. Optimal cloning of mixed Gaussian states

    NASA Astrophysics Data System (ADS)

    Guţă, Mădălin; Matsumoto, Keiji

    2006-09-01

    We construct the optimal one to two cloning transformation for the family of displaced thermal equilibrium states of a harmonic oscillator, with a fixed and known temperature. The transformation is Gaussian and it is optimal with respect to the figure of merit based on the joint output state and norm distance. The proof of the result is based on the equivalence between the optimal cloning problem and that of optimal amplification of Gaussian states which is then reduced to an optimization problem for diagonal states of a quantum oscillator. A key concept in finding the optimum is that of stochastic ordering which plays a similar role in the purely classical problem of Gaussian cloning. The result is then extended to the case of n to m cloning of mixed Gaussian states.

  16. Cloning and characterization of new bioluminescent proteins

    NASA Astrophysics Data System (ADS)

    Szent-Gyorgyi, Christopher; Ballou, Byron T.; Dagnal, Erich; Bryan, Bruce

    1999-07-01

    Over the past two years Prolume has undertaken a comprehensive program to clone luciferases and associated 'green fluorescent proteins' (GFPs) from marine animals that use coelenterazine as the luciferin. To data we have cloned several bioluminescent proteins, including two novel copepod luciferases and two anthozoan GFPs. These four proteins have sequences that differ greatly form previously cloned analogous proteins; the sequence diversity apparently is due to independent evolutionary origins and unusual evolutionary constraints. Thus coelenterazine-based bioluminescent systems may also manifest a variety of useful properties. We discuss form this taxonomic perspective the initial biochemical and spectral characterization of our cloned proteins. Emphasis is placed on the anthozoan luciferase-GFP systems, whose efficient resonance energy transfer has elicited much current interest.

  17. Optimal cloning of mixed Gaussian states

    SciTech Connect

    Guta, Madalin; Matsumoto, Keiji

    2006-09-15

    We construct the optimal one to two cloning transformation for the family of displaced thermal equilibrium states of a harmonic oscillator, with a fixed and known temperature. The transformation is Gaussian and it is optimal with respect to the figure of merit based on the joint output state and norm distance. The proof of the result is based on the equivalence between the optimal cloning problem and that of optimal amplification of Gaussian states which is then reduced to an optimization problem for diagonal states of a quantum oscillator. A key concept in finding the optimum is that of stochastic ordering which plays a similar role in the purely classical problem of Gaussian cloning. The result is then extended to the case of n to m cloning of mixed Gaussian states.

  18. Self-regulating the effortful "social dos".

    PubMed

    Cortes, Kassandra; Kammrath, Lara K; Scholer, Abigail A; Peetz, Johanna

    2014-03-01

    In the current research, we explored differences in the self-regulation of the personal dos (i.e., engaging in active and effortful behaviors that benefit the self) and in the self-regulation of the social dos (engaging in those same effortful behaviors to benefit someone else). In 6 studies, we examined whether the same trait self-control abilities that predict task persistence on personal dos would also predict task persistence on social dos. That is, would the same behavior, such as persisting through a tedious and attentionally demanding task, show different associations with trait self-control when it is framed as benefitting the self versus someone else? In Studies 1-3, we directly compared the personal and social dos and found that trait self-control predicted self-reported and behavioral personal dos but not social dos, even when the behaviors were identical and when the incentives were matched. Instead, trait agreeableness--a trait linked to successful self-regulation within the social domain--predicted the social dos. Trait self-control did not predict the social dos even when task difficulty increased (Study 4), but it did predict the social don'ts, consistent with past research (Studies 5-6). The current studies provide support for the importance of distinguishing different domains of self-regulated behaviors and suggest that social dos can be successfully performed through routes other than traditional self-control abilities. (PsycINFO Database Record (c) 2014 APA, all rights reserved).

  19. Endangered wolves cloned from adult somatic cells.

    PubMed

    Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Yuda, Fibrianto; Kim, Hye Jin; Hwang, Woo Suk; Hossein, Mohammad Shamim; Kim, Joung Joo; Shin, Nam Shik; Kang, Sung Keun; Lee, Byeong Chun

    2007-01-01

    Over the world, canine species, including the gray wolf, have been gradually endangered or extinct. Many efforts have been made to recover and conserve these canids. The aim of this study was to produce the endangered gray wolf with somatic cell nuclear transfer (SCNT) for conservation. Adult ear fibroblasts from a female gray wolf (Canis lupus) were isolated and cultured in vitro as donor cells. Because of limitations in obtaining gray wolf matured oocytes, in vivo matured canine oocytes obtained by flushing the oviducts from the isthmus to the infundibulum were used. After removing the cumulus cells, the oocyte was enucleated, microinjected, fused with a donor cell, and activated. The reconstructed cloned wolf embryos were transferred into the oviducts of the naturally synchronized surrogate mothers. Two pregnancies were detected by ultrasonography at 23 days of gestation in recipient dogs. In each surrogate dog, two fetal sacs were confirmed by early pregnancy diagnosis at 23 days, but only two cloned wolves were delivered. The first cloned wolf was delivered by cesarean section on October 18, 2005, 60 days after embryo transfer. The second cloned wolf was delivered on October 26, 2005, at 61 days postembryo transfer. Microsatellite analysis was performed with genomic DNA from the donor wolf, the two cloned wolves, and the two surrogate female recipients to confirm the genetic identity of the cloned wolves. Analysis of 19 microsatellite loci confirmed that the cloned wolves were genetically identical to the donor wolf. In conclusion, we demonstrated live birth of two cloned gray wolves by nuclear transfer of wolf somatic cells into enucleated canine oocyte, indicating that SCNT is a practical approach for conserving endangered canids.

  20. Lack of association between Gly82Ser, 1704G/T and 2184A/G of RAGE gene polymorphisms and retinopathy susceptibility in Malaysian diabetic patients.

    PubMed

    Ng, Z X; Kuppusamy, U R; Poh, R; Tajunisah, I; Koay, A C A; Fong, K C S; Chua, K H

    2012-03-01

    Diabetic retinopathy is the most common diabetic eye disease, occurring in about 60% of type 2 diabetic patients. Other than known clinical risk factors, the influence of genes has been suggested as part of the development of diabetic retinopathy. We investigated the association of Gly82Ser, 1704G/T and 2184A/G polymorphisms in the RAGE gene with retinopathy in type 2 diabetic patients in Malaysia. Ninety-eight unrelated retinopathy patients and 185 unrelated healthy controls from all over Malaysia were recruited in this study. The allele and genotype frequencies of the three gene polymorphisms were investigated using PCR-RFLP. The allele frequency of the three polymorphisms did not differ significantly between the control and the retinopathy group (P > 0.05). Analysis of the frequency of GA+AA, GT+TT and AG+GG in the retinopathy group did not reveal significant differences (P > 0.05) compared to the control group. We conclude that RAGE gene Gly82Ser, 1704G/T and 2184A/G polymorphisms are not associated with retinopathy development in the Malaysian population.

  1. (GT)n Repeat Polymorphism in Heme Oxygenase-1 (HO-1) Correlates with Clinical Outcome after Myeloablative or Nonmyeloablative Allogeneic Hematopoietic Cell Transplantation

    PubMed Central

    Køllgaard, Tania; Kornblit, Brian; Petersen, Jesper; Klausen, Tobias Wirenfeldt; Mortensen, Bo Kok; Brændstrup, Peter; Sengeløv, Henrik; Høgdall, Estrid; Müller, Klaus; Vindeløv, Lars; Andersen, Mads Hald; thor Straten, Per

    2016-01-01

    Allogeneic hematopoietic cell transplantation (HCT) is a treatment for various hematologic diseases where efficacy of treatment is in part based on the graft versus tumour (GVT) activity of cells in the transplant. The cytoprotective enzyme heme oxygenase-1 (HO-1) is a rate-limiting enzyme in heme degradation and it has been shown to exert anti-inflammatory functions. In humans a (GT)n repeat polymorphism regulates the expression of HO-1. We conducted fragment length analyses of the (GT)n repeat in the promotor region of the gene for HO-1 in DNA from donors and recipients receiving allogeneic myeloablative- (MA) (n = 110) or nonmyeloablative- (NMA-) (n = 250) HCT. Subsequently, we compared the length of the (GT)n repeat with clinical outcome after HCT. We demonstrated that transplants from a HO-1high donor after MA-conditioning (n = 13) is associated with higher relapse incidence at 3 years (p = 0.01, n = 110). In the NMA-conditioning setting transplantation of HO-1low donor cells into HO-1low recipients correlated significantly with decreased relapse related mortality (RRM) and longer progression free survival (PFS) (p = 0.03 and p = 0.008, respectively). Overall, our findings suggest that HO-1 may play a role for the induction of GVT effect after allogeneic HCT. PMID:27997582

  2. CloneAssistant 1.0: a stand-alone software for automated cloning primer design.

    PubMed

    Shao, Chaogang; Meng, Yijun; Lv, Shaolei; Zhong, Wei; Wang, Zheyu; Chen, Ming

    2010-11-01

    "CloneAssistant 1.0" is a stand-alone software compatible with the current Windows operating systems, which can automatically design cloning primers with full consideration of the sequence information of vectors and genes, cloning strategies, the principles of primer design, reading frames, position effects, and enzymatic reaction conditions for users. Five internal XML (extensible markup language) databases [restriction enzymes, plasmids, universal buffers, PCR (polymerase chain reaction) protection bases, and an MCS (multiple cloning site) double digest interference database] were established to serve as the basic support for "CloneAssistant 1.0". The primer pairs designed are sorted according to the difficulty of the follow-up experiments. Once a primer pair is selected by the user, detailed experimental guidance for this primer pair will be provided. In addition, "CloneAssistant 1.0" can be used for restriction map analysis, ORF (open reading frame) finding, sequence alignment and complementary analysis, translation, restriction enzyme and universal buffer queries, and isocaudamer analysis. "CloneAssistant 1.0" makes gene clone design much easier, and it can be freely downloaded from http://bis.zju.edu.cn/clone. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Birth of viable puppies derived from breeding cloned female dogs with a cloned male.

    PubMed

    Park, J E; Hong, S G; Kang, J T; Oh, H J; Kim, M K; Kim, M J; Kim, H J; Kim, D Y; Jang, G; Lee, B C

    2009-09-15

    Since the establishment of production of viable cloned dogs by somatic cell nucleus transfer, great concern has been given to the reproductive abilities of these animals (Canis familiaris). Therefore, we investigated reproductive activity of cloned dogs by (1) performing sperm analysis using computer-assisted sperm analysis and early embryonic development, (2) assessing reproductive cycling by measuring serum progesterone (P4) levels and performing vaginal cytology, and (3) breeding cloned dogs using artificial insemination. Results showed that most parameters of sperm motility in a cloned male dog were within the reference range, and in vivo-matured oocytes from a noncloned female were successfully fertilized by spermatozoa from a cloned male dog and develop normally to the 8-cell stage. Three cloned female dogs displayed normal patterns of P4 levels and morphologic changes of the vaginal epithelium. Two cloned female dogs became pregnant using semen from a cloned male dog and successfully delivered 10 puppies by natural labor. In conclusion, these data demonstrated that both cloned male and female dogs are fertile, and their puppies are currently alive and healthy with normal growth patterns.

  4. A modified version of the digestion-ligation cloning method for more efficient molecular cloning.

    PubMed

    Gao, Song; Li, Yanling; Zhang, Jiannan; Chen, Hongman; Ren, Daming; Zhang, Lijun; An, Yingfeng

    2014-05-15

    Here we describe a modified version of the digestion-ligation approach for efficient molecular cloning. In comparison with the original method, the modified method has the additional steps of gel purification and a second ligation after the first ligation of the linearized vector and DNA insert. During this process, the efficiency and reproducibility could be significantly improved for both stick-end cloning and blunt-end cloning. As an improvement of the very important molecular cloning technique, this method may find a wide range of applications in bioscience and biotechnology.

  5. Emotional reactions to human reproductive cloning.

    PubMed

    May, Joshua

    2016-01-01

    Extant surveys of people's attitudes towards human reproductive cloning focus on moral judgements alone, not emotional reactions or sentiments. This is especially important given that some (especially Leon Kass) have argued against such cloning on the ground that it engenders widespread negative emotions, like disgust, that provide a moral guide. To provide some data on emotional reactions to human cloning, with a focus on repugnance, given its prominence in the literature. This brief mixed-method study measures the self-reported attitudes and emotions (positive or negative) towards cloning from a sample of participants in the USA. Most participants condemned cloning as immoral and said it should be illegal. The most commonly reported positive sentiment was by far interest/curiosity. Negative emotions were much more varied, but anxiety was the most common. Only about a third of participants selected disgust or repugnance as something they felt, and an even smaller portion had this emotion come to mind prior to seeing a list of options. Participants felt primarily interested and anxious about human reproductive cloning. They did not primarily feel disgust or repugnance. This provides initial empirical evidence that such a reaction is not appropriately widespread. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  6. Reproductive cloning combined with genetic modification.

    PubMed

    Strong, C

    2005-11-01

    Although there is widespread opposition to reproductive cloning, some have argued that its use by infertile couples to have genetically related children would be ethically justifiable. Others have suggested that lesbian or gay couples might wish to use cloning to have genetically related children. Most of the main objections to human reproductive cloning are based on the child's lack of unique nuclear DNA. In the future, it may be possible safely to create children using cloning combined with genetic modifications, so that they have unique nuclear DNA. The genetic modifications could be aimed at giving such children genetic characteristics of both members of the couple concerned. Thus, cloning combined with genetic modification could be appealing to infertile, lesbian, or gay couples who seek genetically related children who have genetic characteristics of both members. In such scenarios, the various objections to human reproductive cloning that are based on the lack of genetic uniqueness would no longer be applicable. The author argues that it would be ethically justifiable for such couples to create children in this manner, assuming these techniques could be used safely.

  7. Respiratory disease in neonatal cloned calves.

    PubMed

    Brisville, A C; Fecteau, G; Boysen, S; Dorval, P; Buczinski, S; Blondin, P; Smith, L C

    2011-01-01

    Numerous clinical abnormalities occur in cloned calves during the neonatal period. Describe respiratory diseases affecting cloned calves. Twenty-five cloned Holstein calves. Retrospective clinical study of the cloned calves born at the Veterinary Teaching Hospital, Saint-Hyacinthe, QC. Records of 31 cloned calves were reviewed. Twenty-five records were included. Four stillborn calves and 2 calves euthanized at birth were excluded. Twenty-two calves suffered from respiratory diseases. Nineteen calves received intranasal oxygen treatment (INO). They were tachypneic (78 breaths per minute) and 5 of them were hypoxemic (PaO₂ < 55 mmHg). Two of 19 calves remained hypoxemic despite INO. Thirteen calves were weaned from INO after a median of 70 hours and were discharged at a median of 5 days of age. Nine calves required ventilatory support: 3 from birth and 6 after INO. Five were successfully weaned from the ventilator after a median of 32 hours and were discharged at a median of 8 days of age. Three calves died and 1 was euthanized because of respiratory disease. Necropsy revealed atelectasis, pulmonary congestion, and alveolar damages. Respiratory disease occurs frequently in cloned calves. The most frequent abnormality is hypoxemia because of V/Q mismatch. It is possible to successfully support these calves by INO and mechanical ventilation. Copyright © 2011 by the American College of Veterinary Internal Medicine.

  8. [On the problem of human cloning].

    PubMed

    Smorag, Z

    2001-01-01

    Somatic cell cloning technique in mammals is still not very efficient, but intensive efforts have been made to improve it. Considering the great biological affinity of humans and animals, the cloning technique can in the not too distant future be applied in human cloning and improved to the point of becoming safe. Even when we make such an assumption, I consider it irrational and dangerous to clone the human in order to make their copies (with human cloning for therapeutic purposes being another problem). Life, which is generated by the union of egg cell and spermatozoon is an unforeseeable combination of genetic possibilities, but at the same time it offers a unique chance for the human being, both as an individual and a species. The creation of life by genetic duplication of an already formed individual means a great reduction not only in the biological sense. Action like this is evidence of extreme egocentrism and totalitarian thinking, and its proponents should first answer the question whether they would consider cloning themselves. An answer in the affirmative would help to establish the underlying reasons for their approval.

  9. Cloning: Past, Present, and the Exciting Future. Breakthroughs in Bioscience.

    ERIC Educational Resources Information Center

    Di Berardino, Marie A.

    This document explores the history of cloning by focusing on Dolly the Sheep, one of the first large animal clonings. The disadvantages and advantages of transgenic clones are discussed as well as the future implications of cloning from the perspective of human health. (Contains 10 resources.) (YDS)

  10. The role of the preimplantation geneticist in human cloning.

    PubMed

    Fiddler, M; Pergament, D; Pergament, E

    1999-12-01

    If human cloning is to become a reality, the preimplantation geneticist must be responsible for determining the indications for undertaking cloning and for establishing the risks and benefits of human cloning. The unresolved issue is whether a compelling argument can be made for cloning a human for therapeutic reasons while outweighing legal, moral and ethical objections. At present, 'whole person' cloning does not seem justified under any circumstance, whereas cloning for the replacement of diseased cells, tissues or organ systems, i.e. 'spare parts', seems to be a likely, acceptable application of cloning strategies for humans.

  11. Aberrant gene expression in deceased transgenic cloned calves.

    PubMed

    Zhang, L; Wang, S H; Dai, Y P; Li, N

    2009-05-01

    Several transgenic cloned species have been obtained; however, the efficiency of transgenic cloning remains very low, even lower than cloning. Many experiments have demonstrated abnormal growth and development, and inappropriate gene expression in cloned animals. In this study, we examined the expression of 19 development-related genes in lungs of three normal controls and three aberrant transgenic cloned calves. Results showed in transgenic cloned calves, 84.2% genes had decreased expression levels, however, 5.3% genes had increased levels. This study suggests transgenic cloning and the aberrant expression would cause abnormal growth and development in transgenic cloned calves. To our knowledge, this is the first time that gene expression was examined in transgenic cloned cattle. These findings may have some implications in understanding the low efficiency of the transgenic cloning.

  12. Cloning mice and men: prohibiting the use of iPS cells for human reproductive cloning.

    PubMed

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2010-01-08

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation. Copyright 2010 Elsevier Inc. All rights reserved.

  13. Cloning Mice and Men: Prohibiting the Use of iPS Cells for Human Reproductive Cloning

    PubMed Central

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2014-01-01

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation. PMID:20085739

  14. Update on the First Cloned Dog and Outlook for Canine Cloning.

    PubMed

    Jang, Goo; Lee, ByeongChun

    2015-10-01

    As man's best friend, dogs have an important position in human society. Ten years ago, we reported the first cloned dog, and his birth has raised various scientific issues, such as those related to health, reproduction, and life span. He has developed without any unique health issues. In this article, we summarize and present perspectives on canine cloning.

  15. A DOS Primer for Librarians: Part II.

    ERIC Educational Resources Information Center

    Beecher, Henry

    1990-01-01

    Provides an introduction to DOS commands and strategies for the effective organization and use of hard disks. Functions discussed include the creation of directories and subdirectories, enhanced copying, the assignment of disk drives, and backing up the hard disk. (CLB)

  16. A DOS Primer for Librarians: Part II.

    ERIC Educational Resources Information Center

    Beecher, Henry

    1990-01-01

    Provides an introduction to DOS commands and strategies for the effective organization and use of hard disks. Functions discussed include the creation of directories and subdirectories, enhanced copying, the assignment of disk drives, and backing up the hard disk. (CLB)

  17. High-throughput cloning, expression and purification of glycoside hydrolases using Ligation-Independent Cloning (LIC).

    PubMed

    Camilo, Cesar M; Polikarpov, Igor

    2014-07-01

    Recent advances in DNA sequencing techniques have led to an explosion in the amount of available genome sequencing data and this provided an inexhaustible source of uncharacterized glycoside hydrolases (GH) to be studied both structurally and enzymatically. Ligation-Independent Cloning (LIC), an interesting alternative to traditional, restriction enzyme-based cloning, and commercial recombinatorial cloning, was adopted and optimized successfully for a high throughput cloning, expression and purification pipeline. Using this platform, 130 genes encoding mainly uncharacterized glycoside hydrolases from 13 different organisms were cloned and submitted to a semi-automated protein expression and solubility screening in Escherichia coli, resulting in 73 soluble targets. The high throughput approach proved to be a powerful tool for production of recombinant glycoside hydrolases for further structural and biochemical characterization and confirmed that thioredoxin fusion tag (TRX) is a better choice to increase solubility of recombinant glycoside hydrolases expressed in E. coli, when compared to His-tag alone.

  18. Cloning of a cuticular antigen that contains multiple tandem repeats from the filarial parasite Dirofilaria immitis.

    PubMed Central

    Poole, C B; Grandea, A G; Maina, C V; Jenkins, R E; Selkirk, M E; McReynolds, L A

    1992-01-01

    An unusual antigen composed of tandemly repeated protein units was cloned from the filarial parasite Dirofilaria immitis. The antigen was initially identified by screening a lambda gt11 cDNA library with serum from dogs immunized with irradiated D. immitis third-stage larvae. DNA sequence analysis of the cDNA clone, Di5, revealed a continuous open reading frame composed of two 399-base-pair repeats arranged in tandem. Southern blot analysis of genomic D. immitis DNA showed that the gene coding for Di5 is composed of a tandem array of 25-50 copies of this same 399-base-pair repeat. Antiserum raised against recombinant Di5 protein detected a protein "ladder," from about 14 to greater than 200 kDa with steps approximately 15 kDa apart, on immunoblots of D. immitis extract. Metabolic labeling of adult parasites with [35S]methionine showed that Di5 is synthesized as a large precursor that is subsequently cleaved to produce the ladder-like array. These results suggest that the characteristic ladder is created by proteolytic cleavage of the precursor at the same site in each monomer. The Di5 antigen was localized to the cuticle and hypodermis of adult D. immitis by immunoelectron microscopy. Both male and female parasites were found to release Di5 when cultured in vitro. DNA hybridization analysis demonstrated that Di5 is a member of a gene family present in many filarial parasites that infect both animal and human populations. Images PMID:1631084

  19. Molecular cloning and chromosomal localization of human holocarboxylase synthetase, a gene responsible for biotin dependency

    SciTech Connect

    Suzuki, Y.; Aoki, Y.; Ishida, Y.

    1994-09-01

    Holocarboxylase synthetase (HCS) catalyzes biotin incorporation into various carboxylases that require biotin as a prosthetic group. They are acetyl-CoA carboxylase, a rate-limiting enzyme of fatty acid synthesis; pyruvate carboxylase, a key enzyme of gluconeogenesis; propionyl-CoA carboxylase and 3-methylcrotonyl-CoA carboxylase, enzymes involved in amino acid catabolism. HCS is therefore involved in various metabolic processes and is a key enzyme for biotin utilization by mammalian cells. Deficiency of HCS in man is known to cause biotin-responsive multiple carboxylase deficiency. Isolation of cDNA clones for the enzyme is essential to understand HCS and its deficiency at the molecular level. We purified bovine liver HCS and sequenced its proteolytic peptides. Degenerative oligonucleotide primers were synthesized from the two peptide sequences and used to amplify a putative HCS cDNA fragment from human liver by PCR. Using the amplified DNA fragment as a probe, we screened {lambda}gt10 human liver cDNA library and isolated 12 positive clones. The isolated cDNAs encoded a protein of 726 amino acids with molecular mass of 80,759. The protein contained several sequences identical or similar to those of peptides derived from the bovine liver HCS. The predicted protein had a homologous region with BirA which acts as both a biotin-[acetyl-CoA-carboxylase] ligase and a biotin repressor in E. coli, suggesting a functional relationship between the two proteins. We expressed the protein using pET3 a vector in E. coli (BL21 strain) and raised antiserum against the expressed protein. The antiserum immunoprecipitated HCS activities of human lymphoblasts and bovine liver. A one-base deletion and a missense mutation were found in cells from siblings with HCS deficiency. The human HCS gene was assigned to chromosome 21, region 21q22.1 by fluorescence in situ hybridization analysis.

  20. Benefits and problems with cloning animals.

    PubMed Central

    Smith, L C; Bordignon, V; Babkine, M; Fecteau, G; Keefer, C

    2000-01-01

    Animal cloning is becoming a useful technique for producing transgenic farm animals and is likely to be used to produce clones from valuable adults. Other applications will also undoubtedly be discovered in the near future, such as for preserving endangered breeds and species. Although cloning promises great advantages for commerce and research alike, its outcome is not always certain due to high pregnancy losses and high morbidity and mortality during the neonatal period. Research into the mechanisms involved in the reprogramming of the nucleus is being conducted throughout the world in an attempt to better understand the molecular and cellular mechanisms involved in correcting these problems. Although the cause of these anomalies remains mostly unknown, similar phenotypes have been observed in calves derived through in vitro fertilization, suggesting that culture conditions are involved in these phenomena. In the meantime, veterinarians and theriogenologists have an important role to play in improving the efficiency of cloning by finding treatments to assure normal gestation to term and to develop preventative and curative care for cloned neonates. Images Figure 1. PMID:11143925

  1. Benefits and problems with cloning animals.

    PubMed

    Smith, L C; Bordignon, V; Babkine, M; Fecteau, G; Keefer, C

    2000-12-01

    Animal cloning is becoming a useful technique for producing transgenic farm animals and is likely to be used to produce clones from valuable adults. Other applications will also undoubtedly be discovered in the near future, such as for preserving endangered breeds and species. Although cloning promises great advantages for commerce and research alike, its outcome is not always certain due to high pregnancy losses and high morbidity and mortality during the neonatal period. Research into the mechanisms involved in the reprogramming of the nucleus is being conducted throughout the world in an attempt to better understand the molecular and cellular mechanisms involved in correcting these problems. Although the cause of these anomalies remains mostly unknown, similar phenotypes have been observed in calves derived through in vitro fertilization, suggesting that culture conditions are involved in these phenomena. In the meantime, veterinarians and theriogenologists have an important role to play in improving the efficiency of cloning by finding treatments to assure normal gestation to term and to develop preventative and curative care for cloned neonates.

  2. Cloning and Mutagenesis of a Cytochrome P-450 Locus from Bradyrhizobium japonicum That Is Expressed Anaerobically and Symbiotically

    PubMed Central

    Tully, Raymond E.; Keister, Donald L.

    1993-01-01

    Cytochromes P-450, which in many organisms participate in the metabolism of a variety of endobiotic and xenobiotic substances, are synthesized by symbiotic bacteroids of Bradyrhizobium japonicum. Polyclonal antibodies were raised against two cytochromes P-450 (CYP112 and CYP114) purified from bacteroids. A lambda gt11 expression clone of B. japonicum USDA 110 DNA that reacted with the anti-CYP112 antibody was obtained and was used to screen a library of USDA 110 genomic DNA in pLAFR1 for a clone of the P-450 locus. Forced expression of subclones of the P-450 locus in Escherichia coli produced polypeptides that reacted with either the anti-CYP112 antibody or the anti-CYP114 antibody; no cross-reactivity was evident. A Western blot (immunoblot) analysis showed that neither protein was present in free-living aerobically grown B. japonicum cells, but that both proteins were present in cells grown anaerobically, as well as in bacteroids. A mutant strain disrupted in the CYP112 locus produced neither CYP112 nor CYP114, indicating that the mutation was polar for CYP114. The mutant produced effective nodules on soybeans, even though the bacteroids contained no detectable P-450. This suggests that the cytochromes P-450 which we examined are not involved in an essential symbiotic function. Images PMID:16349113

  3. Tissue-specific expression and cDNA cloning of small nuclear ribonucleoprotein-associated polypeptide N

    SciTech Connect

    McAllister, G.; Amara, S.G.; Lerner, M.R. )

    1988-07-01

    Sera from some patients with systemic lupus erythematosus and other autoimmune diseases have antibodies against nuclear antigens. An example is anti-Sm sera, which recognize proteins associated with small nuclear RNA molecules (small nuclear ribonucleoprotein (snRNP) particles). In this paper anti-Sm sera were used to probe immunoblots of various rat tissues. A previously unidentified M{sub r} 28,000 polypeptide was recognized by these anti-Sm sera. This polypeptide, referred to as N, is expressed in a tissue-specific manner, being most abundant in rat brain, less so in heart, and undetectable in the other tissues examined. Immunoprecipitation experiments using antibodies directed against the cap structure of small nuclear RNAs have demonstrated that N is a snRNP-associated polypeptide. Anti-Sm serum was also used to isolate a partial cDNA clone ({lambda}rb91) from a rat brain phage {lambda}gt11 cDNA expression library. A longer cDNA clone was obtained by rescreening the library with {lambda}rb91. In vitro transcription and subsequent translation of this subcloned, longer insert (pGMA2) resulted in a protein product with the same electrophoretic and immunological properties as N, confirming that pGMA2 encodes N. The tissue distribution of N and the involvement of snRNP particles in nuclear pre-mRNA processing may imply a role for N in tissue-specific pre-mRNA splicing.

  4. Cloning, expression and applicability of thermo-alkali-stable xylanase of Geobacillus thermoleovorans in generating xylooligosaccharides from agro-residues.

    PubMed

    Verma, Digvijay; Satyanarayana, T

    2012-03-01

    A xylanase gene (xyl-gt) of 1.224 kbp was cloned from the extremely thermophilic bacterium Geobacillus thermoleovorans that encodes a protein containing 408 amino acid residues. Eight conserved regions (signature sequences) of GH family 10 xylanases have been found in the xylanase. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the recombinant strain produced xylanase titer of 270 U mg(-1) which is 27-fold higher than the wild strain. It is optimally active at 80°C and pH 8.5 with a high thermostability over broad range of pH (6-12) and temperature (40-100°C). The end products of the hydrolysis of birch wood xylan and agro-residues included xylobiose, xylotriose, xylotetraose and xylopentaose. The xylanase of G. thermoleovorans is one of the rare xylanases that exhibits thermo-alkali-stability, and thus, it is a suitable candidate for pre-bleaching of paper pulps and generating xylooligosaccharides from agro-residues for use as prebiotics.

  5. Genomic cloning, characterization and statistical analysis of an antitumor-analgesic peptide from Chinese scorpion Buthus martensii Karsch.

    PubMed

    Cui, Yong; Liu, Yanfeng; Chen, Qiqing; Zhang, Rong; Song, Yongbo; Jiang, Zhuopu; Wu, Chunfu; Zhang, Jinghai

    2010-09-01

    The genomic DNA sequence encoding an antitumor-analgesic peptide was amplified from the genome of Chinese scorpion Buthus martensii Karsch (BmKAGAP), then cloned and sequenced. An intron, with a high A + T content (61.6%), splits a glycine codon near the end of the precursor signal peptide and the consensus GT/AG splice junction was identified in the BmKAGAP gene. Using PCR amplification, we confirmed the identity of our cloned cDNA, and found that the BmKAGAP gene contained an intron of 506 bp in length, which was almost identical to that of the characterized scorpion sodium channel ligands in size, consensus junctions, putative branch point and A + T content. This is the first report of using a statistical method for Chinese scorpion B. martensii Karsch genomic sequence analysis, involving the extraction of some putative transcription regulatory factors. Moreover, it establishes a theoretical foundation for studying the relationship between scorpion evolution, gene expression and protein function.

  6. [Cloning and structural analysis of MLP in the silkworm, Bombyx mori].

    PubMed

    Liu, Yan; Niu, Bao-Long; Weng, Hong-Biao; Shen, Wei-Feng; He, Li-Hua; Qi, Xiao-Peng; Meng, Zhi-Qi

    2007-09-01

    The LIM domain is found in a wide variety of eukaryotic proteins that regulate gene expression and cell differentiation during development. Muscle LIM protein (MLP) gene in Bombyx mori has been cloned by blasting its EST database and PCR test in present report. The resulting sequence covers 2 327 bp of cDNA (GenBank accession No. DQ311195). It has a complete open reading fragment and encodes a 494 amino acid protein. Genomic DNA sequence contains 11 exons and 10 introns, with intron splicing following the GT-AG rule. M.W. and PI of the predicted MLP in Bombyx mori are 53.03 kDa and 8.29 respectively. A single LIM domain linked to a glyscine-rich region is found in a previously deposited LIM protein (AAR23823) in Bombyx mori. MLP identified in this report encodes a protein with five tandem LIM-glycine modules. The two LIM proteins could be produced by alternative splicing and both are probably involved in muscle cell differentiation. This work provides foundation for further research on the in vivo function of MLP.

  7. Cloning and sequence analysis of candidate human natural killer-enhancing factor genes

    SciTech Connect

    Shau, H.; Butterfield, L.H.; Chiu, R.; Kim, A.

    1994-12-31

    A cytosol factor from human red blood cells enhances natural killer (NK) activity. This factor, termed NK-enhancing factor (NKEF), is a protein of 44000 M{sub r} consisting of two subunits of equal size linked by disulfide bonds. NKEF is expressed in the NK-sensitive erythroleukemic cell line K562. Using an antibody specific for NKEF as a probe for immunoblot screening, we isolated several clones from a {lambda}gt11 cDNA library of K562. Additional subcloning and sequencing revealed that the candidate NKEF cDNAs fell into one of two categories of closely related but non-identical genes, referred to as NKEF A and B. They are 88% identical in amino acid sequence and 71% identical in nucleotide sequence. Southern blot analysis suggests that there are two to three NKEF family members in the genome. Analysis of predicted amino acid sequences indicates that both NKEF A and B are cytosol proteins with several phosphorylation sites each, but that they have no glycosylation sites. They are significantly homologous to several other proteins from a wide variety of organisms ranging from prokaryotes to mammals, especially with regard to several well-conserved motifs within the amino acid sequences. The biological functions of these proteins in other species are mostly unknown, but some of them were reported to be induced by oxidative stress. Therefore, as well as for immunoregulation of NK activity, NKEF may be important for cells in coping with oxidative insults. 32 refs., 3 figs.

  8. Molecular cloning, functional expression, and chromosomal localization of mouse hepatocyte nuclear factor 1

    SciTech Connect

    Kuo, C.J.; Conley, P.B.; Hsieh, Chihlin; Francke, U.; Crabtree, G.R. )

    1990-12-01

    The homeodomain-containing transcription factor hepatocyte nuclear factor 1 (HNF-1) most likely plays and essential role during liver organogenesis by transactivating a family of {gt}15 predominantly hepatic genes. The authors have isolated cDNA clones encoding mouse HNF-1 and expressed them in monkey COS cells and in the human T-cell line Jurkat, producing HNF-1 DNA-binding activity as well as transactivation of reporter constructs containing multimerized NHF-1 binding sites. In addition, the HNF-1 gene was assigned by somatic cell hybrids and recombinant inbred strain mapping to mouse chromosome 5 near Bcd-1 and to human chromosome 12 region q22-qter, revealing a homologous chromosome region in these two species. The presence of HNF-1 mRNA in multiple endodermal tissues (liver, stomach, intestine) suggests that HNF-1 may constitute an early marker for endodermal, rather than hepatocyte, differentiation. Further, that HNF-1 DNA-binding and transcriptional activity can be conferred by transfecting the HNF-1 cDNA into several cell lines indicates that it is sufficient to activate transcription in the context of ubiquitously expressed factors.

  9. Molecular cloning, encoding sequence, and expression of vaccinia virus nucleic acid-dependent nucleoside triphosphatase gene.

    PubMed Central

    Rodriguez, J F; Kahn, J S; Esteban, M

    1986-01-01

    A rabbit poxvirus genomic library contained within the expression vector lambda gt11 was screened with polyclonal antiserum prepared against vaccinia virus nucleic acid-dependent nucleoside triphosphatase (NTPase)-I enzyme. Five positive phage clones containing from 0.72- to 2.5-kilobase-pair (kbp) inserts expressed a beta-galactosidase fusion protein that was reactive by immunoblotting with the NTPase-I antibody. Hybridization analysis allowed the location of this gene within the vaccinia HindIIID restriction fragment. From the known nucleotide sequence of the 16-kbp vaccinia HindIIID fragment, we identified a region that contains a 1896-base open reading frame coding for a 631-amino acid protein. Analysis of the complete sequence revealed a highly basic protein, with hydrophilic COOH and NH2 termini, various hydrophobic domains, and no significant homology to other known proteins. Translational studies demonstrate that NTPase-I belongs to a late class of viral genes. This protein is highly conserved among Orthopoxviruses. Images PMID:3025846

  10. Microenvironmental pH-modified solid dispersions to enhance the dissolution and bioavailability of poorly water-soluble weakly basic GT0918, a developing anti-prostate cancer drug: preparation, characterization and evaluation in vivo.

    PubMed

    Yang, Meiyan; He, Shaolong; Fan, Yunzhou; Wang, Yuli; Ge, Zhenzhong; Shan, Li; Gong, Wei; Huang, Xiaoli; Tong, Youzhi; Gao, Chunsheng

    2014-11-20

    The aim of the present work was to design a pH-modified solid dispersion (pH(M)-SD) that can improve the dissolution and bioavailability of poorly water-soluble weakly basic GT0918, a developing anti-prostate cancer drug. To select the appropriate acidifiers, a solubility test was carried out first. Solid dispersions (SDs) containing GT0918 and polyvinylpyrrolidone (PVP) were prepared using a solvent evaporation method and were characterized using dissolution studies in different media. The solid states of the SDs were investigated using scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC) and Fourier transformed infrared spectroscopy (FTIR). The in vivo pharmacokinetics of the pH(M)-SDs tablets were also studied in beagle dogs compared to the conventional tablets. The optimized pH(M)-SD (GT0918/PVP/citric acid, 1:2:2 weight ratio) exhibited a significant improvement in the dissolution behavior compared to both the physical mixture and the binary SDs. Solid-state characterization revealed that the amorphous formation of GT0918 in the SDs and the strong H-bonding were only found in the pH(M)-SDs containing citric acid. Furthermore, the GT0918-loaded pH(M)-SD tablets showed a higher AUC and a lower tmax compared to the conventional tablets. Accordingly, the pH(M)-SD might be an efficient route for enhancing the dissolution and bioavailability of poorly water-soluble GT0918.

  11. Cloning quantum entanglement in arbitrary dimensions

    SciTech Connect

    Karpov, E.; Navez, P.; Cerf, N.J.

    2005-10-15

    We have found a quantum cloning machine that optimally duplicates the entanglement of a pair of d-dimensional quantum systems prepared in an arbitrary isotropic state. It maximizes the entanglement of formation contained in the two copies of any maximally entangled input state, while preserving the separability of unentangled input states. Moreover, it cannot increase the entanglement of formation of isotropic states. For large d, the entanglement of formation of each clone tends to one-half the entanglement of the input state, which corresponds to a classical behavior. Finally, we investigate a local entanglement cloner, which yields entangled clones with one-fourth the input entanglement in the large-d limit.

  12. Bac clones generated from sheared dna

    SciTech Connect

    Osoegawa, Kazutoyo; Vessere, Gery M.; Shu, Chung Li; Hoskins,Roger A.; Abad, Jose P.; de Pablos, Beatriz; Villasante, Alfredo; deJong, Pieter J.

    2006-08-09

    BAC libraries generated from restriction-digested genomic DNA display representational bias and lack some sequences. To facilitate completion of genome projects, procedures have been developed to create BACs from DNA physically sheared to create fragments extending up to 200kb. The DNA fragments were repaired to create blunt ends and ligated to a new BAC vector. This approach has been tested by generating BAC libraries from Drosophila DNA, with insert lengths of 50 kb to 150 kb. The libraries lack chimeric clone problems as determined by mapping paired BAC-end sequences of one library to the D. melanogaster genome sequence. The utility of ''sheared'' libraries was demonstrated by closure of a previous clone gap and by isolation of clones from telomeric regions, which were notably absent from previous Drosophila BAC libraries.

  13. Scientific hazards of human reproductive 'cloning'.

    PubMed

    Young, Lorraine E

    2003-05-01

    The scientific and clinical professional societies and associations covering the remit of Human Fertility are unanimously opposed to human reproductive 'cloning'. This article describes the main scientific objections to human reproductive 'cloning'. Data collected from numerous studies in a range of animal species indicate a high incidence of fetal defects, a stillbirth rate typically of more than 90% and a lack of adequate information on postnatal development. These concerns are exacerbated by misconceptions about the current ability to screen preimplantation embryos for 'cloning-induced' defects. Scientists and clinicians are sometimes treated with mistrust in the eyes of the public and media over such issues, perhaps because scientific information is not as well communicated as it might be. The duty of reproductive specialists is to convey the limits of their knowledge on this issue to the public and policymakers.

  14. Human reproductive cloning and reasons for deprivation.

    PubMed

    Jensen, D A

    2008-08-01

    Human reproductive cloning provides the possibility of genetically related children for persons for whom present technologies are ineffective. I argue that the desire for genetically related children is not, by itself, a sufficient reason to engage in human reproductive cloning. I show this by arguing that the value underlying the desire for genetically related children implies a tension between the parent and the future child. This tension stems from an instance of a deprivation and violates a general principle of reasons for deprivation. Alternative considerations, such as a right to procreative autonomy, do not appear helpful in making the case for human reproductive cloning merely on the basis of the desire for genetically related children.

  15. Dogs cloned from adult somatic cells.

    PubMed

    Lee, Byeong Chun; Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Yuda, Fibrianto; Kim, Hye Jin; Hossein, M Shamim; Shamim, M Hossein; Kim, Jung Ju; Kang, Sung Keun; Schatten, Gerald; Hwang, Woo Suk

    2005-08-04

    Several mammals--including sheep, mice, cows, goats, pigs, rabbits, cats, a mule, a horse and a litter of three rats--have been cloned by transfer of a nucleus from a somatic cell into an egg cell (oocyte) that has had its nucleus removed. This technology has not so far been successful in dogs because of the difficulty of maturing canine oocytes in vitro. Here we describe the cloning of two Afghan hounds by nuclear transfer from adult skin cells into oocytes that had matured in vivo. Together with detailed sequence information generated by the canine-genome project, the ability to clone dogs by somatic-cell nuclear transfer should help to determine genetic and environmental contributions to the diverse biological and behavioural traits associated with the many different canine breeds.

  16. Cloning whole bacterial genomes in yeast

    PubMed Central

    Benders, Gwynedd A.; Noskov, Vladimir N.; Denisova, Evgeniya A.; Lartigue, Carole; Gibson, Daniel G.; Assad-Garcia, Nacyra; Chuang, Ray-Yuan; Carrera, William; Moodie, Monzia; Algire, Mikkel A.; Phan, Quang; Alperovich, Nina; Vashee, Sanjay; Merryman, Chuck; Venter, J. Craig; Smith, Hamilton O.; Glass, John I.; Hutchison, Clyde A.

    2010-01-01

    Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation. PMID:20211840

  17. Human cloning: three mistakes and an alternative.

    PubMed

    Baylis, Françoise

    2002-06-01

    The current debate on the ethics of cloning humans is both uninspired and uninspiring. In large measure this is because of mistakes that permeate the discourse, including the mistake of thinking that cloning technology is strictly a reproductive technology when it is used to create whole beings. As a result, the challenge this technology represents regarding our understanding of ourselves and the species to which we belong typically is inappropriately downplayed or exaggerated. This has meant that important (albeit disquieting) societal issues and species-type concerns have not been fully explored. This paper, intended as a corrective, suggests that we take an alternate view of human cloning as both an enhancement and a reproductive technology. This proposed shift in the framework for analysis counters the current narrow framing of the issues and introduces new questions about the prospect of modifying the species.

  18. Therapeutic cloning: from consequences to contradiction.

    PubMed

    Coors, Marilyn

    2002-06-01

    The British Parliament legalized therapeutic cloning in December 2000 despite opposition from the European Union. The watershed event in Parliament's move was the active and unprecedented government support for the generation and destruction of human embryonic life merely as a means of medical advancement. This article contends that the utilitarian analysis of this procedure is necessary to identify the real world risks of therapeutic cloning but insufficient to identify the breach of defensible ethical limits that this procedure represents. A value-oriented approach to Kantian ethics demonstrates that the utilitarian endorsement of therapeutic cloning entails a contradiction of the necessity of human vulnerability and a faulty valuation of the human embryo. The concern is that a narrow utilitarian focus ultimately commodifies human embryonic life and preferences outcomes as the sole determinant of moral value.

  19. Cloning whole bacterial genomes in yeast.

    PubMed

    Benders, Gwynedd A; Noskov, Vladimir N; Denisova, Evgeniya A; Lartigue, Carole; Gibson, Daniel G; Assad-Garcia, Nacyra; Chuang, Ray-Yuan; Carrera, William; Moodie, Monzia; Algire, Mikkel A; Phan, Quang; Alperovich, Nina; Vashee, Sanjay; Merryman, Chuck; Venter, J Craig; Smith, Hamilton O; Glass, John I; Hutchison, Clyde A

    2010-05-01

    Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation.

  20. Cloning of Gaussian states by linear optics

    SciTech Connect

    Olivares, Stefano; Paris, Matteo G. A.; Andersen, Ulrik L.

    2006-06-15

    We analyze in details a scheme for cloning of Gaussian states based on linear optical components and homodyne detection recently demonstrated by Andersen et al. [Phys. Rev. Lett. 94, 240503 (2005)]. The input-output fidelity is evaluated for a generic (pure or mixed) Gaussian state taking into account the effect of nonunit quantum efficiency and unbalanced mode mixing. In addition, since in most quantum information protocols the covariance matrix of the set of input states is not perfectly known, we evaluate the average cloning fidelity for classes of Gaussian states with the degree of squeezing and the number of thermal photons being only partially known.

  1. Photonic Programmable Tele-Cloning Network

    NASA Astrophysics Data System (ADS)

    Li, Wei; Chen, Ming-Cheng

    2016-06-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling.

  2. Animal cloning by somatic cell nuclear transfer.

    PubMed

    Smith, Lawrence C; Yoo, Jae-Gyu

    2009-01-01

    Animal cloning is becoming increasingly useful for its applications in biological inquiry and for its potential use in pharmaceutical, medical, and agricultural fields. Due to the complexity of the numerous steps required in reconstructing oocytes by nuclear transfer, detailed protocols are required to minimize the developmental damages inflicted during these manipulations and to standardize procedures across laboratories. Moreover, because oogenesis and early embryogenesis differ widely among mammalian species, it is essential that protocols be adapted according to each species concerned. Our objective here is to detail the protocols that have been most successful in producing laboratory and domestic animal clones.

  3. Cloning of the complete Mycoplasma pneumoniae genome.

    PubMed Central

    Wenzel, R; Herrmann, R

    1989-01-01

    The complete genome of Mycoplasma pneumoniae was cloned in an ordered library consisting of 34 overlapping or adjacent cosmids, one plasmid and two lambda phages. The genome size was determined by adding up the sizes of either the individual unique EcoRI restriction fragments of the gene bank or of the XhoI fragments of genomic M. pneumoniae DNA. The values from these calculations, 835 and 849 kbp, are in good agreement. An XhoI restriction map was constructed by identifying adjacent DNA fragments by probing with selected cosmid clones. Images PMID:2506532

  4. Photonic Programmable Tele-Cloning Network

    PubMed Central

    Li, Wei; Chen, Ming-Cheng

    2016-01-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling. PMID:27353838

  5. Reproduction: widespread cloning in echinoderm larvae.

    PubMed

    Eaves, Alexandra A; Palmer, A Richard

    2003-09-11

    Asexual reproduction by free-living invertebrate larvae is a rare and enigmatic phenomenon and, although it is known to occur in sea stars and brittle stars, it has not been detected in other echinoderms despite more than a century of intensive study. Here we describe spontaneous larval cloning in three species from two more echinoderm classes: a sea cucumber (Holothuroidea), a sand dollar and a sea urchin (Echinoidea). Larval cloning may therefore be an ancient ability of echinoderms and possibly of deutero-stomes - the group that includes echinoderms, acorn worms, sea squirts and vertebrates.

  6. Photonic Programmable Tele-Cloning Network.

    PubMed

    Li, Wei; Chen, Ming-Cheng

    2016-06-29

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling.

  7. A novel NF1 frame-shift mutation (c.702_703delGT) in a Chinese family with neurofibromatosis type 1.

    PubMed

    Cai, S P; Fan, N; Chen, J; Xia, Z L; Wang, Y; Zhou, X M; Yin, Y; Wen, T L; Xia, Q J; Liu, X Y; Wang, H Y

    2014-07-24

    This study aimed to characterize the clinical features of a Chinese pedigree with neurofibromatosis type 1 (NF1) and to identify mutations in the NF1 gene. In this three-generation family containing 8 members, 5 had been diagnosed with NF1 and the others were asymptomatic. All members of the family underwent complete medical examinations. Molecular genetic analyses were performed on all subjects included in the study. All exons of NF1 were amplified by polymerase chain reaction, sequenced, and compared with a reference database. Possible changes in function of the protein induced by amino acid variants were predicted by bioinformatic analysis. In this family, the 5 patients presented different clinical phenotypes, but all manifested typical café-au-lait macules. One novel frame-shift mutation, c.702_703delGT, in exon 7 of NF1 was identified in all affected family members, but not in the unaffected family members or in 102 normal controls. This mutation generates a premature stop codon at amino acid position 720. Additionally, a synonymous mutation c.702 G>A was found in 3 family members, including 2 affected and 1 normal individuals. In conclusion, our study suggests that a novel c.702_703delGT frame-shift mutation in NF1 is likely to be responsible for the pathogenesis of NF1 in this family. To the best of our knowledge, it is the first time that a c.702_703delGT mutation has been identified in a family with neurofibromatosis type 1.

  8. EGFR gene polymorphisms -216G>T and -191C>A are risk markers for gastric cancer in Mexican population.

    PubMed

    Torres-Jasso, J H; Marín, M E; Santiago-Luna, E; Leoner, J C; Torres, J; Magaña-Torres, M T; Perea, F J; Ibarra, B; Sánchez-López, J Y

    2015-03-13

    Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein with tyrosine-kinase activity that plays an important role in multiple cellular functions. EGFR overexpression has been observed in several types of tumors and it is significantly associated with disease stage, survival, prognosis, and progression of cancer. The polymorphisms -216G>T, -191C>A, and (CA)n first intervening sequence (IVS1) have been related to EGFR overexpression and have been studied in several types of cancer, but not in gastric cancer (GC). The aim of this study was to determine the association of these 3 polymorphisms and GC. Genomic DNA from 68 GC patients and 102 healthy blood donors were analyzed. Polymorphisms were identified by DNA-sequencing (-216G>T and -191C>A) and GeneScan (CA)n IVS1. The results showed that the distribution of the -216G>T and -191C>A genotypes differed between groups (P < 0.05). The odds ratio for the -216TT genotype was 4.59 (95% confidence interval = 1.55-13.54, P < 0.05) and 10.71 (95% confidence interval = 2.31-49.59, P < 0.05) for the -191AA genotype, both in a recessive model. The genotype and allele distributions of the (CA)n IVS1 repeat was similar in both groups. In conclusion, the -216TT and -191AA genotypes and GA haplotype of the EGFR gene were found to be associated with an increased risk of gastric cancer in a Mexican population.

  9. GT3X+ accelerometer placement affects the reliability of step-counts measured during running and pedal-revolution counts measured during bicycling.

    PubMed

    Gatti, Anthony A; Stratford, Paul W; Brenneman, Elora C; Maly, Monica R

    2016-01-01

    Accelerometers provide a measure of step-count. Reliability and validity of step-count and pedal-revolution count measurements by the GT3X+ accelerometer, placed at different anatomical locations, is absent in the literature. The purpose of this study was to investigate the reliability and validity of step and pedal-revolution counts produced by the GT3X+ placed at different anatomical locations during running and bicycling. Twenty-two healthy adults (14 men and 8 women) completed running and bicycling activity bouts (5 minutes each) while wearing 6 accelerometers: 2 each at the waist, thigh and shank. Accelerometer and video data were collected during activity. Excellent reliability and validity were found for measurements taken from accelerometers mounted at the waist and shank during running (Reliability: intraclass correlation (ICC) ≥ 0.99; standard error of measurement (SEM) ≤1.0 steps; Pearson ≥ 0.99) and at the thigh and shank during bicycling (Reliability: ICC ≥ 0.99; SEM ≤1.0 revolutions; Pearson ≥ 0.99). Excellent reliability was found between measurements taken at the waist and shank during running (ICC ≥ 0.98; SEM ≤1.6 steps) and between measurements taken at the thigh and shank during bicycling (ICC ≥ 0.99; SEM ≤1.0 revolutions). These data suggest that the GT3X+ can be used for measuring step-count during running and pedal-revolution count during bicycling. Only shank placement is recommended for both activities.

  10. Estrogen inhibition of norepinephrine responsiveness is initiated at the plasma membrane of GnRH-producing GT1-7 cells.

    PubMed

    Morales, Araceli; Gonzalez, Miriam; Marin, Raquel; Diaz, Mario; Alonso, Rafael

    2007-07-01

    The modulatory action of estradiol (E2) on the GnRH network can be exerted indirectly on presynaptic neurons or directly on estrogen receptors (ERs) located within GnRH hypothalamic neurons. Using the GnRH-producing GT1-7 cell line, we have investigated whether E2 is able to modify the response of these cells to norepinephrine (NE) stimulation. A 48-h exposure of GT1-7 cells to 10 nM E2 reduced NE-induced cAMP accumulation. However, 15-min exposure was enough to induce this inhibitory action, provided that a hormone-free period of 48 h after steroid treatment was allowed. Furthermore, this effect was mimicked by E2 coupled to (E-BSA), indicating that it may be exerted through a membrane-mediated mechanism. In addition, competition experiments using E-BSA coupled to fluorescein isothiocyanate (FITC) revealed the presence of cell membrane-binding sites for E2. Binding of E-BSA coupled to FITC was blocked by preincubation of cells with either E2, antiestrogen ICI 182 780, or tamoxifen. Moreover, fluorescence staining of non-permeabilized cells with antibodies against receptors alpha and beta confirmed the presence of both receptor subtypes at the cell membrane. To determine the nature of the ER involved in this response, specific agonists for ERalpha 4,4',4''-(4-propyl-[1H]pyrazole-1,3,5-triyl)tris-phenol (PPT) and ERbeta 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN) were used. Since PPT, but not DPN, reproduced the effect of E2, it is suggested that estrogen-induced modulatory action on NE responsiveness was mediated by the ERalpha isoform. Taken together, these results indicate that E2 modulates the adrenergic sensitivity of GT1-7 cells by a mechanism compatible with the activation of membrane-associated ERs.

  11. Short (GT)n microsatellite repeats in the heme oxygenase-1 gene promoter are associated with antioxidant and anti-inflammatory status in Mexican pediatric patients with sepsis.

    PubMed

    Vázquez-Armenta, Gabriela; González-Leal, Natalia; J Vázquez-de la Torre, Mayra; Muñoz-Valle, José Francisco; Ramos-Márquez, Martha E; Hernández-Cañaveral, Iván; Plascencia-Hernández, Arturo; Siller-López, Fernando

    2013-01-01

    An adequate immune and antioxidant response is a key to the resolution of sepsis. Heme oxygenase-1 (HMOX1) is a stress protein with a polymorphic (GT)n repeat in its gene promoter that regulates its expression in response to oxidative injury, such as that present in sepsis. HMOX1 is the rate-limiting enzyme of heme degradation, and the heme breakdown products, CO, Fe, and bilirubin, are considered to be biologically active metabolites with direct or indirect antioxidant and anti-inflammatory properties. In this study, we investigated the inflammatory and antioxidant response and the relationship with the HMOX1 levels and HMOX1 polymorphism in Mexican septic pediatric patients. In a case-control pilot study, we enrolled 64 septic patients and 72 hospitalized control patients without a diagnosis of sepsis. DNA extracted from buffy coat was genotyped for HMOX1 (GT)n polymorphism by PCR and markers of antioxidant and inflammatory status were quantified in plasma by analysis of the oxygen radical absorbance capacity (ORAC), protein carbonyl (PC), interleukin (IL) 6, IL10, and HMOX1 levels. In septic children, oxidative and inflammatory markers were elevated, and HMOX1 levels were positively correlated with IL10 levels. Genotypic and allelic distribution of HMOX1 polymorphism showed no difference between groups. HMOX1 short-allele septic carriers (< 25 GT repeats) presented favorable ORAC, PC and IL10 levels. This study confirms that an active response against pediatric sepsis involves the expression of HMOX1 and IL10, suggesting that the high antioxidant status associated with HMOX1 short-allele septic carriers might provide a beneficial environment for sepsis resolution.

  12. Serum ASAT, ALAT, ALP, LD, GT, and CK determined in the Cobas-Bio centrifugal analyser by the methods of the Scandinavian Committee on Enzymes.

    PubMed

    Izquierdo, J M; Sotorrío, P; Alvarez-Uría, J; Estrada, J M; Quirós, A

    1982-04-01

    The recommended methods of the Scandinavian Committee on Enzymes [4, 5, 6, 7, 8] have been applied to the Cobas-Bio centrifugal analyser. Reagents and serum volumes were scaled down and final molarities were kept equal. Serum volumes in microliters were as follows: ASAT 30, ALAT 30, ALP 3, LD 5, GT 20, and CK 10. Including the dead space of the sample cup, the volume needed to perform all six tests was 113 microliter. Within-run and between-run precision (CV%) were as follows: ASAT 1.32 and 1.95, ALAT 1.68 and 2.93, ALP 1.56 and 3.10, LD 1.63 and 4.44, GT 0.81 and 2.23, and CK 1.02 and 1.94. Mean deviations (%) from target values of two commercial sera were as follows: ASAT -0.3 and -0.4, ALAT -0.4 and -2.2, ALP -1.8 and -7.3, LD 0.4 and -6.2, GT 13.9 and -10.7, and CK -4.9 and -1.3. Results of all the methods correlated well with those obtained with their respective manual methods. Analytical time for 28 samples of each analyte was 10 min, apart from CK which was 14 min. Reagent cost per sample was 0.6, 0.9, 0.1, 0.3, 0.9, and 26 US cents respectively. All reagents were prepared in the laboratory, except those for CK which were bought from J.T. Baker (Phillipsburgh, NJ, USA). In conclusion, the methods keep the features of the manual methods but they are more precise and practicable, much faster and cheaper, and use minimal amounts of sera more convenient for paediatric work.

  13. Sensitivity of an Elekta iView GT a-Si EPID model to delivery errors for pre-treatment verification of IMRT fields.

    PubMed

    Herwiningsih, Sri; Hanlon, Peta; Fielding, Andrew

    2014-12-01

    A Monte Carlo model of an Elekta iViewGT amorphous silicon electronic portal imaging device (a-Si EPID) has been validated for pre-treatment verification of clinical IMRT treatment plans. The simulations involved the use of the BEAMnrc and DOSXYZnrc Monte Carlo codes to predict the response of the iViewGT a-Si EPID model. The predicted EPID images were compared to the measured images obtained from the experiment. The measured EPID images were obtained by delivering a photon beam from an Elekta Synergy linac to the Elekta iViewGT a-Si EPID. The a-Si EPID was used with no additional build-up material. Frame averaged EPID images were acquired and processed using in-house software. The agreement between the predicted and measured images was analyzed using the gamma analysis technique with acceptance criteria of 3 %/3 mm. The results show that the predicted EPID images for four clinical IMRT treatment plans have a good agreement with the measured EPID signal. Three prostate IMRT plans were found to have an average gamma pass rate of more than 95.0 % and a spinal IMRT plan has the average gamma pass rate of 94.3 %. During the period of performing this work a routine MLC calibration was performed and one of the IMRT treatments re-measured with the EPID. A change in the gamma pass rate for one field was observed. This was the motivation for a series of experiments to investigate the sensitivity of the method by introducing delivery errors, MLC position and dosimetric overshoot, into the simulated EPID images. The method was found to be sensitive to 1 mm leaf position errors and 10 % overshoot errors.

  14. Mutual interaction of kisspeptin, estrogen and bone morphogenetic protein-4 activity in GnRH regulation by GT1-7 cells.

    PubMed

    Terasaka, Tomohiro; Otsuka, Fumio; Tsukamoto, Naoko; Nakamura, Eri; Inagaki, Kenichi; Toma, Kishio; Ogura-Ochi, Kanako; Glidewell-Kenney, Christine; Lawson, Mark A; Makino, Hirofumi

    2013-12-05

    Reproduction is integrated by interaction of neural and hormonal signals converging on hypothalamic neurons for controlling gonadotropin-releasing hormone (GnRH). Kisspeptin, the peptide product of the kiss1 gene and the endogenous agonist for the GRP54 receptor, plays a key role in the regulation of GnRH secretion. In the present study, we investigated the interaction between kisspeptin, estrogen and BMPs in the regulation of GnRH production by using mouse hypothalamic GT1-7 cells. Treatment with kisspeptin increased GnRH mRNA expression and GnRH protein production in a concentration-dependent manner. The expression levels of kiss1 and GPR54 were not changed by kisspeptin stimulation. Kisspeptin induction of GnRH was suppressed by co-treatment with BMPs, with BMP-4 action being the most potent for suppressing the kisspeptin effect. The expression of kisspeptin receptor, GPR54, was suppressed by BMPs, and this effect was reversed in the presence of kisspeptin. It was also revealed that BMP-induced Smad1/5/8 phosphorylation and Id-1 expression were suppressed and inhibitory Smad6/7 was induced by kisspeptin. In addition, estrogen induced GPR54 expression, while kisspeptin increased the expression levels of ERα and ERβ, suggesting that the actions of estrogen and kisspeptin are mutually enhanced in GT1-7 cells. Moreover, kisspeptin stimulated MAPKs and AKT signaling, and ERK signaling was functionally involved in the kisspeptin-induced GnRH expression. BMP-4 was found to suppress kisspeptin-induced GnRH expression by reducing ERK signaling activity. Collectively, the results indicate that the axis of kisspeptin-induced GnRH production is bi-directionally controlled, being augmented by an interaction between ERα/β and GPR54 signaling and suppressed by BMP-4 action in GT1-7 neuron cells.

  15. Diversity of trematode genetic clones within amphipods and the timing of same-clone infections.

    PubMed

    Keeney, Devon B; Waters, Jonathan M; Poulin, Robert

    2007-03-01

    The genetic diversity of trematodes within second intermediate hosts has important implications for the evolution of trematode populations as these hosts are utilized after the parasites reproduce asexually within first intermediate hosts and before sexual reproduction within definitive hosts. We characterised the genetic clonal diversity of the marine trematode Maritrema novaezealandensis within amphipod (Paracalliope novizealandiae) second intermediate hosts using four to six microsatellite loci to determine if multiple copies of identical trematode clones existed within naturally infected amphipods. To determine the relative timing of infections by identical clones within hosts, trematode metacercariae were assigned to six developmental stages and the stages of identical clones were compared. The genotypes of 306 trematodes were determined from 44 amphipods each containing more than one trematode. Six pairs of identical trematode clones were recovered in total (representing five amphipods: 11% of amphipods with greater than one trematode) and all pairs of clones belonged to the same developmental stage. This suggests that identical clone infections are effectively synchronous. A general decrease in the number of metacercariae recovered, prevalence, and mean intensity of infection for each subsequent developmental stage coupled with large numbers of metacercariae (>9) only being recovered from recent infections, supports the occurrence of post-infection amphipod mortality and/or within-host trematode mortality. Taken together, our results indicate that natural infections are characterised by high genetic diversity, but that amphipods also periodically encounter "batches" of genetically identical clones, potentially setting the stage for interactions within and between clonal groups inside the host.

  16. Whole genome comparison of donor and cloned dogs.

    PubMed

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-10-21

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic cell nuclear transfer produced an almost identical genome. The whole genome sequence data of donor and cloned dogs can provide a resource for further investigations on epigenetic contributions in phenotypic differences.

  17. Thinking about cloning: a reply to Judith Thomson.

    PubMed

    Blackford, R

    2001-11-01

    Opponents of human cloning typically argue for the prohibition of therapeutic cloning and a permanent prohibition of reproductive cloning, even if a safe cloning technology should become available. In a recent article in this journal, "Legal and Ethical Problems of Human Cloning" (2000) 8 JLM 31, Judith Thomson develops an ethico-legal analysis that would justify prohibitions or restrictions on both therapeutic and reproductive cloning, irrespective of any safety issue. This article criticises Thomson's analysis in detail and suggests, in particular, that it relies upon an intellectually unacceptable understanding of personhood.

  18. Whole genome comparison of donor and cloned dogs

    PubMed Central

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-01-01

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic cell nuclear transfer produced an almost identical genome. The whole genome sequence data of donor and cloned dogs can provide a resource for further investigations on epigenetic contributions in phenotypic differences. PMID:24141358

  19. Patterns of p53 G-->T transversions in lung cancers reflect the primary mutagenic signature of DNA-damage by tobacco smoke.

    PubMed

    Hainaut, P; Pfeifer, G P

    2001-03-01

    It is unquestionable that the major cause of lung cancer is cigarette smoking. p53 mutations are common in lung cancers from smokers but less common in non-smokers. A large fraction of the p53 mutations in lung cancers are G-->T transversions, a type of mutation that is infrequent in other tumors aside from hepatocellular carcinoma. Previous studies have indicated that there is a good correlation between G-->T transversion hotspots in lung cancers and sites of preferential formation of polycyclic aromatic hydrocarbon (PAH) adducts along the p53 gene. The origin of p53 mutations in lung cancer has been questioned by recent reports suggesting that there are no significant differences in p53 mutation spectra between smokers and non-smokers and between lung cancers and non-lung cancers [S.N. Rodin and A.S. Rodin (2000) Human lung cancer and p53: The interplay between mutagenesis and selection. P:roc. Natl Acad. Sci. USA, 97, 12244-12249]. We have re-assessed these issues by using the latest update of the p53 mutation database of the International Agency for Research on Cancer (14 051 entries) as well as recent data from the primary literature on non-smokers. We come to the conclusion that the p53 mutation spectra are different between smokers and non-smokers and that this difference is highly statistically significant (G-->T transversions are 30 versus 10%; P < 0.0001, chi2 test). A similar difference is seen between lung cancers and non-lung cancers. At a number of mutational hotspots common to all cancers, a large fraction of the mutations are G-->T transversions in lung cancers but are almost exclusively G-->A transitions in non-lung cancers. Our data reinforce the notion that p53 mutations in lung cancers can be attributed to direct DNA damage from cigarette smoke carcinogens rather than to selection of pre-existing endogenous mutations.

  20. GT-repeat polymorphism in the heme oxygenase-1 gene promoter and the risk of carotid atherosclerosis related to arsenic exposure

    PubMed Central

    2010-01-01

    Background Arsenic is a strong stimulus of heme oxygenase (HO)-1 expression in experimental studies in response to oxidative stress caused by a stimulus. A functional GT-repeat polymorphism in the HO-1 gene promoter was inversely correlated to the development of coronary artery disease in diabetics and development of restenosis following angioplasty in patients. The role of this potential vascular protective factor in carotid atherosclerosis remains unclear. We previously reported a graded association of arsenic exposure in drinking water with an increased risk of carotid atherosclerosis. In this study, we investigated the relationship between HO-1 genetic polymorphism and the risk of atherosclerosis related to arsenic. Methods Three-hundred and sixty-seven participants with an indication of carotid atherosclerosis and an additional 420 participants without the indication, which served as the controls, from two arsenic exposure areas in Taiwan, a low arsenic-exposed Lanyang cohort and a high arsenic-exposed LMN cohort, were studied. Carotid atherosclerosis was evaluated using a duplex ultrasonographic assessment of the extracranial carotid arteries. Allelic variants of (GT)n repeats in the 5'-flanking region of the HO-1 gene were identified and grouped into a short (S) allele (< 27 repeats) and long (L) allele (≥ 27 repeats). The association of atherosclerosis and the HO-1 genetic variants was assessed by a logistic regression analysis, adjusted for cardiovascular risk factors. Results Analysis results showed that arsenic's effect on carotid atherosclerosis differed between carriers of the class S allele (OR 1.39; 95% CI 0.86-2.25; p = 0.181) and non-carriers (OR 2.65; 95% CI 1.03-6.82; p = 0.044) in the high-exposure LMN cohort. At arsenic exposure levels exceeding 750 μg/L, difference in OR estimates between class S allele carriers and non-carriers was borderline significant (p = 0.051). In contrast, no such results were found in the low-exposure Lanyang