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Sample records for system facilitating cloning

  1. Generation of a vector system facilitating cloning of DMBT1 variants and recombinant expression of functional full-length DMBT1

    DEFF Research Database (Denmark)

    End, Caroline; Lyer, Stefan; Renner, Marcus

    2005-01-01

    Deleted in malignant brain tumours 1 (DMBT1) codes for a approximately 340kDa glycoprotein with highly repetitive scavenger receptor cysteine-rich (SRCR) domains. DMBT1 was implicated in cancer, defence against viral and bacterial infections, and differentiation of epithelial cells. Recombinant...... expression and purification of DMBT1 is an essential step for systematic standardized functional research and towards the evaluation of its therapeutical potential. So far, DMBT1 is obtained from natural sources such as bronchioalveolar lavage or saliva, resulting in time consuming sample collection, low...... yields, and protein preparations which may substantially vary due to differential processing and genetic polymorphism, all of which impedes functional research on DMBT1. Cloning of DMBT1 cDNAs is hampered because of the size and the 13 highly homologous SRCR exons. In this study, we report on the setup...

  2. Clone tag detection in distributed RFID systems.

    Science.gov (United States)

    Kamaludin, Hazalila; Mahdin, Hairulnizam; Abawajy, Jemal H

    2018-01-01

    Although Radio Frequency Identification (RFID) is poised to displace barcodes, security vulnerabilities pose serious challenges for global adoption of the RFID technology. Specifically, RFID tags are prone to basic cloning and counterfeiting security attacks. A successful cloning of the RFID tags in many commercial applications can lead to many serious problems such as financial losses, brand damage, safety and health of the public. With many industries such as pharmaceutical and businesses deploying RFID technology with a variety of products, it is important to tackle RFID tag cloning problem and improve the resistance of the RFID systems. To this end, we propose an approach for detecting cloned RFID tags in RFID systems with high detection accuracy and minimal overhead thus overcoming practical challenges in existing approaches. The proposed approach is based on consistency of dual hash collisions and modified count-min sketch vector. We evaluated the proposed approach through extensive experiments and compared it with existing baseline approaches in terms of execution time and detection accuracy under varying RFID tag cloning ratio. The results of the experiments show that the proposed approach outperforms the baseline approaches in cloned RFID tag detection accuracy.

  3. Clone tag detection in distributed RFID systems

    Science.gov (United States)

    Kamaludin, Hazalila; Mahdin, Hairulnizam

    2018-01-01

    Although Radio Frequency Identification (RFID) is poised to displace barcodes, security vulnerabilities pose serious challenges for global adoption of the RFID technology. Specifically, RFID tags are prone to basic cloning and counterfeiting security attacks. A successful cloning of the RFID tags in many commercial applications can lead to many serious problems such as financial losses, brand damage, safety and health of the public. With many industries such as pharmaceutical and businesses deploying RFID technology with a variety of products, it is important to tackle RFID tag cloning problem and improve the resistance of the RFID systems. To this end, we propose an approach for detecting cloned RFID tags in RFID systems with high detection accuracy and minimal overhead thus overcoming practical challenges in existing approaches. The proposed approach is based on consistency of dual hash collisions and modified count-min sketch vector. We evaluated the proposed approach through extensive experiments and compared it with existing baseline approaches in terms of execution time and detection accuracy under varying RFID tag cloning ratio. The results of the experiments show that the proposed approach outperforms the baseline approaches in cloned RFID tag detection accuracy. PMID:29565982

  4. Cloning

    Science.gov (United States)

    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 ...

  5. Ultra-low background DNA cloning system.

    Directory of Open Access Journals (Sweden)

    Kenta Goto

    Full Text Available Yeast-based in vivo cloning is useful for cloning DNA fragments into plasmid vectors and is based on the ability of yeast to recombine the DNA fragments by homologous recombination. Although this method is efficient, it produces some by-products. We have developed an "ultra-low background DNA cloning system" on the basis of yeast-based in vivo cloning, by almost completely eliminating the generation of by-products and applying the method to commonly used Escherichia coli vectors, particularly those lacking yeast replication origins and carrying an ampicillin resistance gene (Amp(r. First, we constructed a conversion cassette containing the DNA sequences in the following order: an Amp(r 5' UTR (untranslated region and coding region, an autonomous replication sequence and a centromere sequence from yeast, a TRP1 yeast selectable marker, and an Amp(r 3' UTR. This cassette allowed conversion of the Amp(r-containing vector into the yeast/E. coli shuttle vector through use of the Amp(r sequence by homologous recombination. Furthermore, simultaneous transformation of the desired DNA fragment into yeast allowed cloning of this DNA fragment into the same vector. We rescued the plasmid vectors from all yeast transformants, and by-products containing the E. coli replication origin disappeared. Next, the rescued vectors were transformed into E. coli and the by-products containing the yeast replication origin disappeared. Thus, our method used yeast- and E. coli-specific "origins of replication" to eliminate the generation of by-products. Finally, we successfully cloned the DNA fragment into the vector with almost 100% efficiency.

  6. Expression systems for cloned xenobiotic transporters

    International Nuclear Information System (INIS)

    Pritchard, John B.; Miller, David S.

    2005-01-01

    One challenge of modern biology is to be able to match genes and their encoded proteins with events at the molecular, cellular, tissue, and organism levels, and thus, provide a multi-level understanding of gene function and dysfunction. How well this can be done for xenobiotic transporters depends on a knowledge of the genes expressed in the tissue, the cellular locations of the gene products (do they function for uptake or efflux?), and our ability to match substrates with transporters using information obtained from cloned transporters functioning in heterologous expression systems. Clearly, making a rational choice of expression system to use for the characterization and study of cloned xenobiotic transporters is a critical part of study design. This choice requires well-defined goals, as well as an understanding of the strengths and weaknesses of candidate expression systems

  7. Cloning

    Science.gov (United States)

    ... been cloned from somatic cells include: cat, deer, dog, horse, mule, ox, rabbit and rat. In addition, ... to make copies of animals with the potential benefits for the fields of medicine and agriculture. For ...

  8. A dual-site gateway cloning system for simultaneous cloning of two genes for plant transformation.

    Science.gov (United States)

    Aboulela, Mostafa; Tanaka, Yuji; Nishimura, Kohji; Mano, Shoji; Kimura, Tetsuya; Nakagawa, Tsuyoshi

    2017-07-01

    Analyses of the subcellular localization of proteins and protein-protein interaction networks are essential to uncover the molecular basis of diverse biological processes in plants. To this end, we have created a Gateway cloning-compatible vector system, named dual-site (DS) Gateway cloning system to allow simple cloning of two expression cassettes in a binary vector and to express them simultaneously in plant cells. In the DS Gateway cloning system, (i) a moderate constitutive nopaline synthase promoter (Pnos), which is much suitable for localization analysis, is used to guide each expression cassette, (ii) four series of vectors with different plant resistance markers are established, (iii) N-terminal fusion with 6 fluorescent proteins and 7 epitope tags is available, (iv) both N- and C-terminal fusions with split enhanced yellow fluorescent protein (EYFP) are possible for efficient detection of protein-protein interactions using a bimolecular fluorescence complementation (BiFC) assay. The usefulness of the DS Gateway cloning system has been demonstrated by the analysis of the expression and the subcellular localization patterns of two Golgi proteins in stable expression system using A. thaliana, and by the analyses of interactions between subunits of coat protein complex II (COPII) both in transient and stable expression systems using Japanese leek and A. thaliana, respectively. The DS Gateway cloning system provides a multipurpose, efficient expression tool in gene function analyses and especially suitable for investigating interactions and subcellular localization of two proteins in living plant cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Full-length RecE enhances linear-linear homologous recombination and facilitates direct cloning for bioprospecting.

    Science.gov (United States)

    Fu, Jun; Bian, Xiaoying; Hu, Shengbaio; Wang, Hailong; Huang, Fan; Seibert, Philipp M; Plaza, Alberto; Xia, Liqiu; Müller, Rolf; Stewart, A Francis; Zhang, Youming

    2012-05-01

    Functional analysis of genome sequences requires methods for cloning DNA of interest. However, existing methods, such as library cloning and screening, are too demanding or inefficient for high-throughput application to the wealth of genomic data being delivered by massively parallel sequencing. Here we describe direct DNA cloning based on the discovery that the full-length Rac prophage protein RecE and its partner RecT mediate highly efficient linear-linear homologous recombination mechanistically distinct from conventional recombineering mediated by Redαβ from lambda phage or truncated versions of RecET. We directly cloned all ten megasynthetase gene clusters (each 10–52 kb in length) from Photorhabdus luminescens into expression vectors and expressed two of them in a heterologous host to identify the metabolites luminmycin A and luminmide A/B. We also directly cloned cDNAs and exactly defined segments from bacterial artificial chromosomes. Direct cloning with full-length RecE expands the DNA engineering toolbox and will facilitate bioprospecting for natural products.

  10. Cloning systems for Rhodococcus and related bacteria

    Science.gov (United States)

    Finnerty, W.R.; Singer, M.E.

    1990-08-28

    A plasmid transformation system for Rhodococcus was developed using an Escherichia coli-Rhodococcus shuttle plasmid. Rhodococcus sp. H13-A contains three cryptic indigenous plasmids, designated pMVS100, pMVS200 and pMVS300, of 75, 19.5 and 13.4 kilobases (Kb), respectively. A 3.8 Kb restriction fragment of pMVS300 was cloned into pIJ30, a 6.3 Kb pBR322 derivative, containing the E. coli origin of replication (ori) and ampicillin resistance determinant (bla) as well as a Streptomyces gene for thiostrepton resistance, tsr. The resulting 10.1 Kb recombinant plasmid, designated pMVS301, was isolated from E. coli DH1 (pMVS301) and transformed into Rhodococcus sp. AS-50, a derivative of strain H13-A, by polyethylene glycol-assisted transformation of Rhodococcus protoplasts and selection for thiostrepton-resistant transformants. This strain was deposited with the ATCC on Feb. 1, 1988 and assigned ATCC 53719. The plasmid contains the Rhodococcus origin of replication. The plasmid and derivatives thereof can therefore be used to introduce nucleic acid sequences to and from Rhodococcus for subsequent expression and translation into protein. The isolated origin of replication can also be used in the construction of new vectors. 2 figs.

  11. Cloning systems for Rhodococcus and related bacteria

    Science.gov (United States)

    Finnerty, William R.; Singer, Mary E.

    1990-01-01

    A plasmid transformation system for Rhodococcus was developed using an Escherichia coli-Rhodococcus shuttle plasmid. Rhodococcus sp. H13-A contains three cryptic indigenous plasmids, designated pMVS100, pMVS200 and pMVS300, of 75, 19.5 and 13.4 kilobases (Kb), respectively. A 3.8 Kb restriction fragment of pMVS300 was cloned into pIJ30, a 6.3 Kb pBR322 derivative, containing the E. coli origin of replication (ori) and ampicillin resistance determinant (bla) as well as a Streptomyces gene for thiostrepton resistance, tsr. The resulting 10.1 Kb recombinant plasmid, designated pMVS301, was isolated from E. coli DH1 (pMVS301) and transformed into Rhodococcus sp. AS-50, a derivative of strain H13-A, by polyethylene glycol-assisted transformation of Rhodococcus protoplasts and selection for thiostrepton-resistant transformants. This strain was deposited with the ATCC on Feb. 1, 1988 and assigned ATCC 53719. The plasmid contains the Rhodococcus origin of replication. The plasmid and derivatives thereof can therefore be used to introduce nucleic acid sequences to and from Rhodococcus for subsequent expression and translation into protein. The isolated origin of replication can also be used in the construction of new vectors.

  12. Extreme Environments Facilitate Hybrid Superiority - The Story of a Successful Daphnia galeata × longispina Hybrid Clone.

    Science.gov (United States)

    Griebel, Johanna; Gießler, Sabine; Poxleitner, Monika; Navas Faria, Amanda; Yin, Mingbo; Wolinska, Justyna

    2015-01-01

    Hybridization within the animal kingdom has long been underestimated. Hybrids have often been considered less fit than their parental species. In the present study, we observed that the Daphnia community of a small lake was dominated by a single D. galeata × D. longispina hybrid clone, during two consecutive years. Notably, in artificial community set-ups consisting of several clones representing parental species and other hybrids, this hybrid clone took over within about ten generations. Neither the fitness assay conducted under different temperatures, or under crowded and non-crowded environments, nor the carrying capacity test revealed any outstanding life history parameters of this hybrid clone. However, under simulated winter conditions (i.e. low temperature, food and light), the hybrid clone eventually showed a higher survival probability and higher fecundity compared to parental species. Hybrid superiority in cold-adapted traits leading to an advantage of overwintering as parthenogenetic lineages might consequently explain the establishment of successful hybrids in natural communities of the D. longispina complex. In extreme cases, like the one reported here, a superior hybrid genotype might be the only clone alive after cold winters. Overall, superiority traits, such as enhanced overwintering here, might explain hybrid dominance in nature, especially in extreme and rapidly changing environments. Although any favoured gene complex in cyclic parthenogens could be frozen in successful clones independent of hybridization, we did not find similarly successful clones among parental species. We conclude that the emergence of the observed trait is linked to the production of novel recombined hybrid genotypes.

  13. Quantum cloning attacks against PUF-based quantum authentication systems

    Science.gov (United States)

    Yao, Yao; Gao, Ming; Li, Mo; Zhang, Jian

    2016-08-01

    With the advent of physical unclonable functions (PUFs), PUF-based quantum authentication systems have been proposed for security purposes, and recently, proof-of-principle experiment has been demonstrated. As a further step toward completing the security analysis, we investigate quantum cloning attacks against PUF-based quantum authentication systems and prove that quantum cloning attacks outperform the so-called challenge-estimation attacks. We present the analytical expression of the false-accept probability by use of the corresponding optimal quantum cloning machines and extend the previous results in the literature. In light of these findings, an explicit comparison is made between PUF-based quantum authentication systems and quantum key distribution protocols in the context of cloning attacks. Moreover, from an experimental perspective, a trade-off between the average photon number and the detection efficiency is discussed in detail.

  14. In vivo blunt-end cloning through CRISPR/Cas9-facilitated non-homologous end-joining

    Science.gov (United States)

    Geisinger, Jonathan M.; Turan, Sören; Hernandez, Sophia; Spector, Laura P.; Calos, Michele P.

    2016-01-01

    The CRISPR/Cas9 system facilitates precise DNA modifications by generating RNA-guided blunt-ended double-strand breaks. We demonstrate that guide RNA pairs generate deletions that are repaired with a high level of precision by non-homologous end-joining in mammalian cells. We present a method called knock-in blunt ligation for exploiting these breaks to insert exogenous PCR-generated sequences in a homology-independent manner without loss of additional nucleotides. This method is useful for making precise additions to the genome such as insertions of marker gene cassettes or functional elements, without the need for homology arms. We successfully utilized this method in human and mouse cells to insert fluorescent protein cassettes into various loci, with efficiencies up to 36% in HEK293 cells without selection. We also created versions of Cas9 fused to the FKBP12-L106P destabilization domain in an effort to improve Cas9 performance. Our in vivo blunt-end cloning method and destabilization-domain-fused Cas9 variant increase the repertoire of precision genome engineering approaches. PMID:26762978

  15. A versatile prokaryotic cloning vector with six dual restriction enzyme sites in the polylinker facilitates efficient subcloning into vectors with unique cloning sites.

    Science.gov (United States)

    Sage, D R; Chillemi, A C; Fingeroth, J D

    1998-09-01

    In large and complex vectors a single restriction enzyme recognition site may be available for introduction of additional DNA requiring the development of linker fragments to create compatible insertion sites. This technology can be time consuming and costly. We describe the construction of a simple phagemid, pSFI, with a polylinker that contains six pairs of dual, rare-cutting, restriction enzyme recognition sites (NotI, SpeI, EcoRV, PstI, SacII, EagI) with multiple unique sites between each pair. This has permitted rapid subcloning of DNA with creation of single flanking restriction enzyme sites. pSFI was used to expedite transfer of viral genes to a LacZ-inducible expression vector and to an adenovirus expression cassette for production of replication-defective virus. The use of this phagemid has facilitated complex vector manipulations and is a valuable adjunct to the family of multifunctional cloning vectors.

  16. Insertion of Introns: A Strategy to Facilitate Assembly of Infectious Full Length Clones

    DEFF Research Database (Denmark)

    Johansen, Ida Elisabeth; Lund, Ole Søgaard

    2008-01-01

    Some DNA fragments are difficult to clone in Escherichia coli by standard methods. It has been speculated that unintended transcription and translation result in expression of proteins that are toxic to the bacteria. This problem is frequently observed during assembly of infectious full-length vi......Some DNA fragments are difficult to clone in Escherichia coli by standard methods. It has been speculated that unintended transcription and translation result in expression of proteins that are toxic to the bacteria. This problem is frequently observed during assembly of infectious full...... the virus sequence in the plant nucleus. The resulting RNA, which enters the cytoplasm, is identical to the virus sequence and can initiate infection...

  17. A modular cloning system for standardized assembly of multigene constructs.

    Directory of Open Access Journals (Sweden)

    Ernst Weber

    Full Text Available The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering.

  18. A Gateway recombination herpesvirus cloning system with negative selection that produces vectorless progeny.

    Science.gov (United States)

    Kunec, Dusan; van Haren, Sandra; Burgess, Shane C; Hanson, Larry A

    2009-01-01

    Crossover recombination based on the lambda phage integration/excision functions enables insertion of a gene of interest into a specific locus by a simple one-step in vitro recombination reaction. Recently, a highly efficient recombination system for targeted mutagenesis, which utilizes lambda phage crossover recombination cloning, has been described for a human herpesvirus 2 bacterial artificial chromosome (BAC). The disadvantages of the system are that it allows only neutral selection (loss of green fluorescent protein) of desired recombinants and that it regenerates herpesvirus progeny containing the BAC sequence inserted in the herpesvirus genome. In this study, the existing channel catfish herpesvirus (CCV) infectious clone (in the form of overlapping fragments) was modified to allow introduction of foreign genes by modified lambda phage crossover recombination cloning. This novel system enables negative and neutral selection and regenerates vectorless herpesvirus progeny. Construction of two CCV mutants expressing lacZ, one from the native CCV ORF5 promoter and the other from the immediate-early cytomegalovirus promoter, demonstrated the efficiency and reliability of this system. This novel cloning system enables rapid incorporation, direct delivery and high-level expression of foreign genes by a herpesvirus. This system has broad utility and could be used to facilitate development of recombinant viruses, viral vectors and better vaccines.

  19. Development of an R4 dual-site (R4DS gateway cloning system enabling the efficient simultaneous cloning of two desired sets of promoters and open reading frames in a binary vector for plant research.

    Directory of Open Access Journals (Sweden)

    Mostafa Aboulela

    Full Text Available Vast numbers of proteins work cooperatively to exert their functions in various cells. In order to understand the functions and molecular mechanisms of these proteins in plants, analyses of transgenic plants that concomitantly express two protein-coding genes are often required. We developed a novel Gateway cloning technology-compatible binary vector system, the R4 dual-site (R4DS Gateway cloning system, which enables the easy and efficient cloning of two desired sets of promoters and open reading frames (ORFs into a binary vector using promoter and ORF entry clones. In this system, C-terminal fusions with 17 kinds of tags including visible reporters and epitope tags are available for each ORF, and selection by four kinds of resistance markers is possible. We verified that the R4DS Gateway cloning system functioned well in Arabidopsis thaliana by observing the expression and localization patterns of fluorescent proteins fused with organelle-targeting signals and driven by stomatal-lineage specific promoters. We also confirmed that the two cloning sites in the R4DS Gateway cloning system were equivalent and independently regulated. The results obtained indicate that the R4DS Gateway cloning system facilitates detailed comparisons of the expression patterns of two promoters as well as co-localization and interaction analyses of two proteins in specific cells in plants.

  20. BTFS: The Border Trade Facilitation System

    Energy Technology Data Exchange (ETDEWEB)

    Phillips, L.R.

    1999-03-18

    The author demonstrates the Border Trade Facilitation System (BTFS), an agent-based bilingual e-commerce system built to expedite the regulation, control, and execution of commercial trans-border shipments during the delivery phase. The system was built to serve maquila industries at the US/Mexican border. The BTFS uses foundation technology developed here at Sandia Laboratories' Advanced Information Systems Lab (AISL), including a distributed object substrate, a general-purpose agent development framework, dynamically generated agent-human interaction via the World-Wide Web, and a collaborative agent architecture. This technology is also the substrate for the Multi-Agent Simulation Management System (MASMAS) proposed for demonstration at this conference. The BTFS executes authenticated transactions among agents performing open trading over the Internet. With the BTFS in place, one could conduct secure international transactions from any site with an Internet connection and a web browser. The BTFS is currently being evaluated for commercialization.

  1. Probabilistic cloning with supplementary information contained in the quantum states of two auxiliary systems

    International Nuclear Information System (INIS)

    Li, Lvjun; Qiu, Daowen

    2007-01-01

    In probabilistic cloning with two auxiliary systems, we consider and compare three different protocols for the success probabilities of cloning. We show that, in certain circumstances, it may increase the success probability to add an auxiliary system to the probabilistic cloning machine having one auxiliary system, but we always can find another cloning machine with one auxiliary system having the same success probability as that with two auxiliary systems

  2. Quantum cloning machines and their implementation in physical systems

    International Nuclear Information System (INIS)

    Wu Tao; Ye Liu; Fang Bao-Long

    2013-01-01

    We review the basic theory of approximate quantum cloning for discrete variables and some schemes for implementing quantum cloning machines. Several types of approximate quantum clones and their expansive quantum clones are introduced. As for the implementation of quantum cloning machines, we review some design methods and recent experimental results. (topical review - quantum information)

  3. Genome modifications and cloning using a conjugally transferable recombineering system

    Directory of Open Access Journals (Sweden)

    Mohammad J Hossain

    2015-12-01

    Full Text Available The genetic modification of primary bacterial disease isolates is challenging due to the lack of highly efficient genetic tools. Herein we describe the development of a modified PCR-based, λ Red-mediated recombineering system for efficient deletion of genes in Gram-negative bacteria. A series of conjugally transferrable plasmids were constructed by cloning an oriT sequence and different antibiotic resistance genes into recombinogenic plasmid pKD46. Using this system we deleted ten different genes from the genomes of Edwardsiella ictaluri and Aeromonas hydrophila. A temperature sensitive and conjugally transferable flp recombinase plasmid was developed to generate markerless gene deletion mutants. We also developed an efficient cloning system to capture larger bacterial genetic elements and clone them into a conjugally transferrable plasmid for facile transferring to Gram-negative bacteria. This system should be applicable in diverse Gram-negative bacteria to modify and complement genomic elements in bacteria that cannot be manipulated using available genetic tools.

  4. Facilitating Adoption of an Electronic Documentation System.

    Science.gov (United States)

    Jones, Nazarine T; Seckman, Charlotte

    2018-01-12

    Best practice recommends the integration of clinical documentation into the hospital electronic health record to support safe, efficient, and timely patient care. A major barrier to successful adoption and optimization of computerized documentation systems is user satisfaction. The purpose of this descriptive, performance improvement initiative was to implement and evaluate user satisfaction with an electronic documentation system to facilitate successful adoption. The Clinical Procedure Flowsheets application was implemented in a geriatric extended care unit of a large healthcare system. Rogers' Diffusion of Innovation and Davis' Technology Acceptance Model were used to guide system adoption and improve user experience of the innovation. The Perceived Usefulness and Perceived Ease of Use questionnaire was distributed to 24 nursing staff working in the unit 9 weeks after implementation. Results indicated that respondents perceived the Clinical Procedure Flowsheets as easy to use and useful in accomplishing their documentation tasks. The overall mean satisfaction score of 72.17 (SD, 12.13) implied a strong level of user acceptance. The positive perception of the nursing staff in the geriatric extended care unit suggests a high probability of system use that can enhance the documentation of patient care. Further research is recommended to evaluate factors related to system adoption and user satisfaction.

  5. Clone-based Data Index in Cloud Storage Systems

    Directory of Open Access Journals (Sweden)

    He Jing

    2016-01-01

    Full Text Available The storage systems have been challenged by the development of cloud computing. The traditional data index cannot satisfy the requirements of cloud computing because of the huge index volumes and quick response time. Meanwhile, because of the increasing size of data index and its dynamic characteristics, the previous ways, which rebuilding the index or fully backup the index before the data has changed, cannot satisfy the need of today’s big data index. To solve these problems, we propose a double-layer index structure that overcomes the throughput limitation of single point server. Then, a clone based B+ tree structure is proposed to achieve high performance and adapt dynamic environment. The experimental results show that our clone-based solution has high efficiency.

  6. Cloning of replication-incompetent herpes simplex viruses as bacterial artificial chromosomes to facilitate development of vectors for gene delivery into differentiated neurons.

    Science.gov (United States)

    Schmeisser, Falko; Weir, Jerry P

    2006-01-01

    We have previously described the adaptation of a tetracycline-regulated system of gene expression for herpes simplex virus (HSV) vectors and demonstrated that such a system was capable of inducible foreign gene expression in irreversibly differentiated neurons. These studies suggested that such gene delivery vectors would be especially useful for studying the neuron in vitro. Here, we describe the cloning of a replication-incompetent HSV vector as a bacterial artificial chromosome (BAC) to facilitate vector construction. Using prokaryotic genetic techniques for allele replacement, we demonstrate the ease of manipulation of the BAC-containing vector, including the construction of vector mutations for which there is no simple phenotypic selection. Such constructions include the insertion of a tetracycline-regulated gene cassette into the UL41 gene for regulated gene expression and the mutation of the UL48 gene to reduce vector toxicity. In addition, HSV vectors cloned as BACs can be sequentially modified to make multiple changes to the vector platform. Finally, using the BAC system, we constructed an HSV vector that expressed an inducible human superoxide dismutase-1 (SOD1) gene for delivery into differentiated human NT-neurons (cells of the human embryonal carcinoma cell line NT2, which differentiate irreversibly into postmitotic neuron-like cells after treatment with retinoic acid). The results indicated that there is appreciable expression of SOD1 from this HSV vector in the presence of doxycycline and that vector-expressed SOD1 interacts with endogenous SOD1. Thus, the BAC system provides a practicable platform for construction and manipulation of HSV vectors that are suitable for gene delivery into postmitotic neurons in vitro.

  7. Heteroduplex DNA mismatch repair system of Streptococcus pneumoniae: cloning and expression of the hexA gene

    International Nuclear Information System (INIS)

    Balganesh, T.S.; Lacks, S.A.

    1985-01-01

    Mutations affecting heteroduplex DNA mismatch repair in Streptococcus pneumoniae were localized in two genes, hexA and hexB, by fractionation of restriction fragments carrying mutant alleles. A fragment containing the hexA4 allele was cloned in the S. pneumoniae cloning system, and the hexA + allele was introduced into the recombinant plasmid by chromosomal facilitation of plasmid transfer. Subcloning localized the functional hexA gene to a 3.5-kilobase segment of the cloned pneumococcal DNA. The product of this gene was shown in Bacillus subtilis minicells to be a polypeptide with an M/sub r/ of 86,000. Two mutant alleles of hexA showed partial expression of the repair system when present in multicopy plasmids. A model for mismatch repair, which depends on the interaction of two protein components to recognize the mismatched base pair and excise a segment of DNA between strand breaks surrounding the mismatch, is proposed

  8. Facilitering

    DEFF Research Database (Denmark)

    Ravn, Ib

    2012-01-01

    Facilitering (af latin facilis: gørbart, let at gøre) er den teknik at gøre det lettere for en forsamlet gruppe mennesker at udrette det, den ønsker. Facilitator er en slags mødeleder eller ordstyrer, der bistår gruppen ved at styre formen på deltagernes samtale og interaktion snarere end indholdet...

  9. Modified-cytosine restriction-system-induced recombinant cloning artefacts in Escherichia coli

    DEFF Research Database (Denmark)

    Williamson, M R; Doherty, J P; Woodcock, D M

    1993-01-01

    We have tested whether, and to what extent, recombinant clones from DNA segments with 5-methylation of cytosines recovered in methylation-restrictive (mcr+) hosts contain mutations. We constructed a model system in which the tetracycline-resistance-encoding gene (tet) from pBR322 was cloned......A+ mcr+ hosts compared with a methylation-tolerant host (mcr-). Of the clones recovered in recA+mcr+ hosts, > 20% of clones had an inactivating mutation in tet. The majority of such mutant clones contained deletions that frequently extended into the unmethylated portion of tet and even into the plasmid...

  10. Time-Efficient Cloning Attacks Identification in Large-Scale RFID Systems

    Directory of Open Access Journals (Sweden)

    Ju-min Zhao

    2017-01-01

    Full Text Available Radio Frequency Identification (RFID is an emerging technology for electronic labeling of objects for the purpose of automatically identifying, categorizing, locating, and tracking the objects. But in their current form RFID systems are susceptible to cloning attacks that seriously threaten RFID applications but are hard to prevent. Existing protocols aimed at detecting whether there are cloning attacks in single-reader RFID systems. In this paper, we investigate the cloning attacks identification in the multireader scenario and first propose a time-efficient protocol, called the time-efficient Cloning Attacks Identification Protocol (CAIP to identify all cloned tags in multireaders RFID systems. We evaluate the performance of CAIP through extensive simulations. The results show that CAIP can identify all the cloned tags in large-scale RFID systems fairly fast with required accuracy.

  11. Extreme Environments Facilitate Hybrid Superiority – The Story of a Successful Daphnia galeata × longispina Hybrid Clone

    Science.gov (United States)

    Griebel, Johanna; Gießler, Sabine; Poxleitner, Monika; Navas Faria, Amanda; Yin, Mingbo; Wolinska, Justyna

    2015-01-01

    Hybridization within the animal kingdom has long been underestimated. Hybrids have often been considered less fit than their parental species. In the present study, we observed that the Daphnia community of a small lake was dominated by a single D. galeata × D. longispina hybrid clone, during two consecutive years. Notably, in artificial community set-ups consisting of several clones representing parental species and other hybrids, this hybrid clone took over within about ten generations. Neither the fitness assay conducted under different temperatures, or under crowded and non-crowded environments, nor the carrying capacity test revealed any outstanding life history parameters of this hybrid clone. However, under simulated winter conditions (i.e. low temperature, food and light), the hybrid clone eventually showed a higher survival probability and higher fecundity compared to parental species. Hybrid superiority in cold-adapted traits leading to an advantage of overwintering as parthenogenetic lineages might consequently explain the establishment of successful hybrids in natural communities of the D. longispina complex. In extreme cases, like the one reported here, a superior hybrid genotype might be the only clone alive after cold winters. Overall, superiority traits, such as enhanced overwintering here, might explain hybrid dominance in nature, especially in extreme and rapidly changing environments. Although any favoured gene complex in cyclic parthenogens could be frozen in successful clones independent of hybridization, we did not find similarly successful clones among parental species. We conclude that the emergence of the observed trait is linked to the production of novel recombined hybrid genotypes. PMID:26448651

  12. An optimized lentiviral vector system for conditional RNAi and efficient cloning of microRNA embedded short hairpin RNA libraries.

    Science.gov (United States)

    Adams, Felix F; Heckl, Dirk; Hoffmann, Thomas; Talbot, Steven R; Kloos, Arnold; Thol, Felicitas; Heuser, Michael; Zuber, Johannes; Schambach, Axel; Schwarzer, Adrian

    2017-09-01

    RNA interference (RNAi) and CRISPR-Cas9-based screening systems have emerged as powerful and complementary tools to unravel genetic dependencies through systematic gain- and loss-of-function studies. In recent years, a series of technical advances helped to enhance the performance of virally delivered RNAi. For instance, the incorporation of short hairpin RNAs (shRNAs) into endogenous microRNA contexts (shRNAmiRs) allows the use of Tet-regulated promoters for synchronous onset of gene knockdown and precise interrogation of gene dosage effects. However, remaining challenges include lack of efficient cloning strategies, inconsistent knockdown potencies and leaky expression. Here, we present a simple, one-step cloning approach for rapid and efficient cloning of miR-30 shRNAmiR libraries. We combined a human miR-30 backbone retaining native flanking sequences with an optimized all-in-one lentiviral vector system for conditional RNAi to generate a versatile toolbox characterized by higher doxycycline sensitivity, reduced leakiness and enhanced titer. Furthermore, refinement of existing shRNA design rules resulted in substantially improved prediction of powerful shRNAs. Our approach was validated by accurate quantification of the knockdown potency of over 250 single shRNAmiRs. To facilitate access and use by the scientific community, an online tool was developed for the automated design of refined shRNA-coding oligonucleotides ready for cloning into our system. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    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

  14. Molecular transformation, gene cloning, and gene expression systems for filamentous fungi

    Science.gov (United States)

    Gold, Scott E.; Duick, John W.; Redman, Regina S.; Rodriguez, Rusty J.

    2001-01-01

    This chapter discusses the molecular transformation, gene cloning, and gene expression systems for filamentous fungi. Molecular transformation involves the movement of discrete amounts of DNA into cells, the expression of genes on the transported DNA, and the sustainable replication of the transforming DNA. The ability to transform fungi is dependent on the stable replication and expression of genes located on the transforming DNA. Three phenomena observed in bacteria, that is, competence, plasmids, and restriction enzymes to facilitate cloning, were responsible for the development of molecular transformation in fungi. Initial transformation success with filamentous fungi, involving the complementation of auxotrophic mutants by exposure to sheared genomic DNA or RNA from wt isolates, occurred with low transformation efficiencies. In addition, it was difficult to retrieve complementing DNA fragments and isolate genes of interest. This prompted the development of transformation vectors and methods to increase efficiencies. The physiological studies performed with fungi indicated that the cell wall could be removed to generate protoplasts. It was evident that protoplasts could be transformed with significantly greater efficiencies than walled cells.

  15. Facilitating Automation Development in Internal Logistics Systems

    OpenAIRE

    Granlund, Anna

    2014-01-01

    The internal logistics system includes all activities connected with managing the flow of materials within the physical limits of a facility. This system is an important part of operations in need of increased focus and continuous improvements. Automation is one possible tool with a previously confirmed great potential to improve internal logistics. Despite this great potential and a growing trend of using automation in the area, internal logistics activities are still not automated to the sa...

  16. Modified-cytosine restriction-system-induced recombinant cloning artefacts in Escherichia coli

    DEFF Research Database (Denmark)

    Williamson, M R; Doherty, J P; Woodcock, D M

    1993-01-01

    We have tested whether, and to what extent, recombinant clones from DNA segments with 5-methylation of cytosines recovered in methylation-restrictive (mcr+) hosts contain mutations. We constructed a model system in which the tetracycline-resistance-encoding gene (tet) from pBR322 was cloned into ...

  17. Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination.

    Science.gov (United States)

    Nikiforuk, Aidan M; Leung, Anders; Cook, Bradley W M; Court, Deborah A; Kobasa, Darwyn; Theriault, Steven S

    2016-10-01

    Viral Infectious clone systems serve as robust platforms to study viral gene or replicative function by reverse genetics, formulate vaccines and adapt a wild type-virus to an animal host. Since the development of the first viral infectious clone system for the poliovirus, novel strategies of viral genome construction have allowed for the assembly of viral genomes across the identified viral families. However, the molecular profiles of some viruses make their genome more difficult to construct than others. Two factors that affect the difficulty of infectious clone construction are genome length and genome complexity. This work examines the available strategies for overcoming the obstacles of assembling the long and complex RNA genomes of coronaviruses and reports one-step construction of an infectious clone system for the Middle East Respiratory Syndrome coronavirus (MERS-CoV) by homologous recombination in S. cerevisiae. Future use of this methodology will shorten the time between emergence of a novel viral pathogen and construction of an infectious clone system. Completion of a viral infectious clone system facilitates further study of a virus's biology, improvement of diagnostic tests, vaccine production and the screening of antiviral compounds. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  18. Academic Cloning.

    Science.gov (United States)

    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…

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

    Science.gov (United States)

    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.

  20. Homologous recombination-mediated cloning and manipulation of genomic DNA regions using Gateway and recombineering systems.

    Science.gov (United States)

    Rozwadowski, Kevin; Yang, Wen; Kagale, Sateesh

    2008-11-17

    Employing genomic DNA clones to characterise gene attributes has several advantages over the use of cDNA clones, including the presence of native transcription and translation regulatory sequences as well as a representation of the complete repertoire of potential splice variants encoded by the gene. However, working with genomic DNA clones has traditionally been tedious due to their large size relative to cDNA clones and the presence, absence or position of particular restriction enzyme sites that may complicate conventional in vitro cloning procedures. To enable efficient cloning and manipulation of genomic DNA fragments for the purposes of gene expression and reporter-gene studies we have combined aspects of the Gateway system and a bacteriophage-based homologous recombination (i.e. recombineering) system. To apply the method for characterising plant genes we developed novel Gateway and plant transformation vectors that are of small size and incorporate selectable markers which enable efficient identification of recombinant clones. We demonstrate that the genomic coding region of a gene can be directly cloned into a Gateway Entry vector by recombineering enabling its subsequent transfer to Gateway Expression vectors. We also demonstrate how the coding and regulatory regions of a gene can be directly cloned into a plant transformation vector by recombineering. This construct was then rapidly converted into a novel Gateway Expression vector incorporating cognate 5' and 3' regulatory regions by using recombineering to replace the intervening coding region with the Gateway Destination cassette. Such expression vectors can be applied to characterise gene regulatory regions through development of reporter-gene fusions, using the Gateway Entry clones of GUS and GFP described here, or for ectopic expression of a coding region cloned into a Gateway Entry vector. We exemplify the utility of this approach with the Arabidopsis PAP85 gene and demonstrate that the expression

  1. Systems design and engineering : facilitating multidisciplinary development projects

    NARCIS (Netherlands)

    Bonnema, Gerrit Maarten; Veenvliet, Karel; Broenink, Johannes F.

    2016-01-01

    As its name implies, the aim of Systems Design and Engineering: Facilitating Multidisciplinary Development Projects is to help systems engineers develop the skills and thought processes needed to successfully develop and implement engineered systems. Such expertise typically does not come through

  2. GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules.

    Directory of Open Access Journals (Sweden)

    Alejandro Sarrion-Perdigones

    Full Text Available Synthetic Biology requires efficient and versatile DNA assembly systems to facilitate the building of new genetic modules/pathways from basic DNA parts in a standardized way. Here we present GoldenBraid (GB, a standardized assembly system based on type IIS restriction enzymes that allows the indefinite growth of reusable gene modules made of standardized DNA pieces. The GB system consists of a set of four destination plasmids (pDGBs designed to incorporate multipartite assemblies made of standard DNA parts and to combine them binarily to build increasingly complex multigene constructs. The relative position of type IIS restriction sites inside pDGB vectors introduces a double loop ("braid" topology in the cloning strategy that allows the indefinite growth of composite parts through the succession of iterative assembling steps, while the overall simplicity of the system is maintained. We propose the use of GoldenBraid as an assembly standard for Plant Synthetic Biology. For this purpose we have GB-adapted a set of binary plasmids for A. tumefaciens-mediated plant transformation. Fast GB-engineering of several multigene T-DNAs, including two alternative modules made of five reusable devices each, and comprising a total of 19 basic parts are also described.

  3. The Flp double cross system a simple efficient procedure for cloning DNA fragments

    Directory of Open Access Journals (Sweden)

    Sadowski Paul D

    2003-07-01

    Full Text Available Abstract Background While conventional cloning methods using restriction enzymes and polynucleotide ligase are adequate for most DNAs, fragments made by the polymerase chain reaction are difficult to clone because the amplifying DNA polymerase tends to add untemplated nucleotides to the 3'-termini of the amplified strands. Conservative site-specific recombinases offer an efficient alternative to conventional cloning methods. Results In this paper I describe the use of the Flp site-specific recombinase for cloning PCR-amplified fragments. A DNA fragment is amplified with primers that contain at their ends inverted target sequences for Flp. Flp readily recombines these fragments in vitro into a vector that also contains two inverted Flp target sequences surrounding the α-complementing region of the lacZ gene of E. coli. The recombinants are conveniently detected as white colonies by the familiar blue/white screening test for lacZ activity. A useful feature of the system is that both orientations of the inserted DNA are usually obtained. If the recipient vector is cut between the two inverted Flp targets, Flp "heals" the double-strand break by inserting a linear fragment flanked by Flp targets. Conclusion This system ("The Flp Double Cross System" should be useful for cloning multiple PCR fragments into many sites in several vectors. It has certain advantages over other available recombinase-based cloning procedures.

  4. Strategies to facilitate the development of uncloned or cloned infectious full-length viral cDNAs: Apple chlorotic leaf spot virus as a case study

    Directory of Open Access Journals (Sweden)

    Marais Armelle

    2011-10-01

    Full Text Available Abstract Background Approaches to simplify and streamline the construction of full-length infectious cDNA clones (FL-cDNAs are needed. Among desirable improvements are the ability to use total nucleic acids (TNA extracts from infected hosts (to bypass viral purification limitations for the direct one-step amplification of large FL-cDNAs, the possibility to inoculate plants with uncloned FL-cDNAs and the simplified cloning of these large molecules. Results Using the 7.55 kb genome of Apple chlorotic leaf spot trichovirus (ACLSV approaches allowing the rapid generation from TNA extracts of FL-cDNAs under the control of the T7 promoter and the successful inoculation of plants using in vitro transcripts obtained from these uncloned amplification products have been developed. We also show that the yeast homologous recombination system permits efficient cloning of FL-cDNAs and the simultaneous one-step tailoring of a ternary Yeast-Escherichia coli-Agrobacterium tumefaciens shuttle vector allowing efficient inoculation of both herbaceous and woody host plants by agroinfiltration. Conclusions The fast and efficient strategies described here should have broad applications, in particular for the study of "difficult" plant viruses, such as those infecting woody hosts, and potentially for other, non plant-infecting viral agents.

  5. Why Clone?

    Science.gov (United States)

    ... the ways in which cloning might be useful. Cloning in Medicine Cloning for medical purposes has the ... people. How might cloning be used in medicine? Cloning animal models of disease Much of what researchers ...

  6. Cloning of observables

    OpenAIRE

    Ferraro, Alessandro; Galbiati, Matteo; Paris, Matteo G. A.

    2005-01-01

    We introduce the concept of cloning for classes of observables and classify cloning machines for qubit systems according to the number of parameters needed to describe the class under investigation. A no-cloning theorem for observables is derived and the connections between cloning of observables and joint measurements of noncommuting observables are elucidated. Relationships with cloning of states and non-demolition measurements are also analyzed.

  7. Cloning of observables

    International Nuclear Information System (INIS)

    Ferraro, Alessandro; Galbiati, Matteo; Paris, Matteo G A

    2006-01-01

    We introduce the concept of cloning for classes of observables and classify cloning machines for qubit systems according to the number of parameters needed to describe the class under investigation. A no-cloning theorem for observables is derived and the connections between cloning of observables and joint measurements of noncommuting observables are elucidated. Relationships with cloning of states and non-demolition measurements are also analysed. (letter to the editor)

  8. Neural mechanism of facilitation system during physical fatigue.

    Directory of Open Access Journals (Sweden)

    Masaaki Tanaka

    Full Text Available An enhanced facilitation system caused by motivational input plays an important role in supporting performance during physical fatigue. We tried to clarify the neural mechanisms of the facilitation system during physical fatigue using magnetoencephalography (MEG and a classical conditioning technique. Twelve right-handed volunteers participated in this study. Participants underwent MEG recording during the imagery of maximum grips of the right hand guided by metronome sounds for 10 min. Thereafter, fatigue-inducing maximum handgrip trials were performed for 10 min; the metronome sounds were started 5 min after the beginning of the handgrip trials. The metronome sounds were used as conditioned stimuli and maximum handgrip trials as unconditioned stimuli. The next day, they were randomly assigned to two groups in a single-blinded, two-crossover fashion to undergo two types of MEG recordings, that is, for the control and motivation sessions, during the imagery of maximum grips of the right hand guided by metronome sounds for 10 min. The alpha-band event-related desynchronizations (ERDs of the motivation session relative to the control session within the time windows of 500 to 700 and 800 to 900 ms after the onset of handgrip cue sounds were identified in the sensorimotor areas. In addition, the alpha-band ERD within the time window of 400 to 500 ms was identified in the right dorsolateral prefrontal cortex (Brodmann's area 46. The ERD level in the right dorsolateral prefrontal cortex was positively associated with that in the sensorimotor areas within the time window of 500 to 700 ms. These results suggest that the right dorsolateral prefrontal cortex is involved in the neural substrates of the facilitation system and activates the sensorimotor areas during physical fatigue.

  9. Understanding Science: Frameworks for using stories to facilitate systems thinking

    Science.gov (United States)

    ElShafie, S. J.; Bean, J. R.

    2017-12-01

    Studies indicate that using a narrative structure for teaching and learning helps audiences to process and recall new information. Stories also help audiences retain specific information, such as character names or plot points, in the context of a broader narrative. Stories can therefore facilitate high-context systems learning in addition to low-context declarative learning. Here we incorporate a framework for science storytelling, which we use in communication workshops, with the Understanding Science framework developed by the UC Museum of Paleontology (UCMP) to explore the application of storytelling to systems thinking. We translate portions of the Understanding Science flowchart into narrative terms. Placed side by side, the two charts illustrate the parallels between the scientific process and the story development process. They offer a roadmap for developing stories about scientific studies and concepts. We also created a series of worksheets for use with the flowcharts. These new tools can generate stories from any perspective, including a scientist conducting a study; a character that plays a role in a larger system (e.g., foraminifera or a carbon atom); an entire system that interacts with other systems (e.g., the carbon cycle). We will discuss exemplar stories about climate change from each of these perspectives, which we are developing for workshops using content and storyboard models from the new UCMP website Understanding Global Change. This conceptual framework and toolkit will help instructors to develop stories about scientific concepts for use in a classroom setting. It will also help students to analyze stories presented in class, and to create their own stories about new concepts. This approach facilitates student metacognition of the learning process, and can also be used as a form of evaluation. We are testing this flowchart and its use in systems teaching with focus groups, in preparation for use in teacher professional development workshops.

  10. Hand-made cloned buffalo (Bubalus bubalis) embryos: comparison of different media and culture systems.

    Science.gov (United States)

    Shah, Riaz A; George, Aman; Singh, Manoj K; Kumar, Dharmendra; Chauhan, Manmohan S; Manik, Radhaysham; Palta, Prabhat; Singla, Suresh K

    2008-12-01

    Hand-made cloning (HMC) has proved to be an efficient alternative to the conventional micromanipulator-based technique in some domestic animal species. This study reports the development of an effective culture system for in vitro culture of zona-free cloned buffalo (Bubalus bubalis) embryos reconstructed using adult skin fibroblast cells as nucleus donor. Cleavage and blastocyst rates observed were 52 and 0% in modified Charles Rosenkrans 2 (mCR2), 61 and 4.6% in modified Synthetic Oviductal Fluid (mSOF), and 82 and 40.3% in Research Vitro Cleave (RVCL; Cook, Australia) medium, respectively. Similarly, higher blastocyst rates (24.5 +/- 4.1%) were observed when zona-free parthenotes were cultured in RVCL medium. Culturing zona-free cloned buffalo embryos on flat surfaces (FS) yielded significantly higher (p WOW) or microdrops (MD). Furthermore, development in WOW was found to be significantly better than MD culture. The quality of HMC blastocysts was examined using differential staining. This study establishes the application of zona-free nuclear transfer procedures for the production of hand-made cloned buffalo embryos and the development of efficient culture system and appropriate media requirements for enhancing their preimplantation development.

  11. Systems Genetics Implicates Cytoskeletal Genes in Oocyte Control of Cloned Embryo Quality

    Science.gov (United States)

    Cheng, Yong; Gaughan, John; Midic, Uros; Han, Zhiming; Liang, Cheng-Guang; Patel, Bela G.; Latham, Keith E.

    2013-01-01

    Cloning by somatic cell nuclear transfer is an important technology, but remains limited due to poor rates of success. Identifying genes supporting clone development would enhance our understanding of basic embryology, improve applications of the technology, support greater understanding of establishing pluripotent stem cells, and provide new insight into clinically important determinants of oocyte quality. For the first time, a systems genetics approach was taken to discover genes contributing to the ability of an oocyte to support early cloned embryo development. This identified a primary locus on mouse chromosome 17 and potential loci on chromosomes 1 and 4. A combination of oocyte transcriptome profiling data, expression correlation analysis, and functional and network analyses yielded a short list of likely candidate genes in two categories. The major category—including two genes with the strongest genetic associations with the traits (Epb4.1l3 and Dlgap1)—encodes proteins associated with the subcortical cytoskeleton and other cytoskeletal elements such as the spindle. The second category encodes chromatin and transcription regulators (Runx1t1, Smchd1, and Chd7). Smchd1 promotes X chromosome inactivation, whereas Chd7 regulates expression of pluripotency genes. Runx1t1 has not been associated with these processes, but acts as a transcriptional repressor. The finding that cytoskeleton-associated proteins may be key determinants of early clone development highlights potential roles for cytoplasmic components of the oocyte in supporting nuclear reprogramming. The transcriptional regulators identified may contribute to the overall process as downstream effectors. PMID:23307892

  12. SCI-Clone/32 - a distributed real time simulation system

    International Nuclear Information System (INIS)

    Wilks, C.F.

    1986-01-01

    Advances in engineering and in particular digital computers has enabled the simulation manufacturers to deliver a realism of a kind undreamt of a decade ago. 32-bit computers ranging in processor power from several hundred thousand instructions per second to many millions are at the heart of each simulator. Gould has pioneered digital computers in simulation with real time systems using shared memory, parallel processors, 64KByte cache, and shadow memory. The market is planning for higher iteration rates, lower life cycle costs, and the development of part task products. These can be met by distributing the tasks amongst nodal computers having a unique architecture for sharing data variables with minimal contention. (Auth.)

  13. Teaching strategic and systems design to facilitate collaboration and learning

    Directory of Open Access Journals (Sweden)

    Andre Liem

    2012-08-01

    Full Text Available As strategic and systems approaches are becoming more relevant in design education when it concerns collaborative projects with the industry, an explicit systems design methodology is needed to structure collaboration and learning among students, educators, and the Norwegian industry. This article describes three alternative studio projects for teaching strategic and systems design with the involvement of Norwegian companies. Besides this, the approaches and fundamental theories of design thinking and reasoning, which are characteristic of these projects, were reflected against each other. In the undergraduate (year 2 systems thinking design studio, the challenge was to train students to understand how system elements are rationally interconnected with their suprasystems and subsystems based on usability and man-machine interactions. In addition to the challenges pertaining to systems thinking, collabora­tive learning and designing based on a mentorship learning concept were introduced in the Vertical Design Studio, which involved second- and third-year students. Concerning the postgraduate fourth-year strategic design projects with the industry, the challenge was to involve Norwegian companies in product planning and goal finding as well as in innovation and design activities and to assess how supportive and receptive these companies were towards radical innovation/diversification. The analysis of completed projects shows that the Norwegian industry is supportive of strategic design but is rather conservative and risk averse when it concerns accepting and implementing radical innovation initiatives. Referring to user-centred and context-based innovation, this article also supports the implementation of a systems approach to facilitate social and hierarchical learning across the second-year systems design studio, second- and third-year vertical studios, and fourth-year strategic design studio.

  14. A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions.

    Science.gov (United States)

    Karna, S L Rajasekhar; Zogaj, Xhavit; Barker, Jeffrey R; Seshu, Janakiram; Dove, Simon L; Klose, Karl E

    2010-11-01

    Comprehensive clone sets representing the entire genome now exist for a large number of organisms. The Gateway entry clone sets are a particularly useful means to study gene function, given the ease of introduction into any Gateway-suitable destination vector. We have adapted a bacterial two-hybrid system for use with Gateway entry clone sets, such that potential interactions between proteins encoded within these clone sets can be determined by new destination vectors. We show that utilizing the Gateway clone sets for Francisella tularensis and Vibrio cholerae, known interactions between F. tularensis IglA and IglB and V. cholerae VipA and VipB could be confirmed with these destination vectors. Moreover, the introduction of unique tags into each vector allowed for visualization of the expressed hybrid proteins via Western immunoblot. This Gateway-suitable bacterial two-hybrid system provides a new tool for rapid screening of protein-protein interactions.

  15. Method and system to facilitate sealing in gas turbines

    Science.gov (United States)

    Morgan, Victor John; Foster, Gregory Thomas; Sarawate, Neelesh Nandkumar

    2017-09-12

    A method and system for sealing between components within a gas turbine is provided. A first recess defined in a first component receives a seal member. A second recess defined in a second component adjacent the first component also receives the seal member. The first and second recesses are located proximate a hot gas path defined through the gas turbine, and define circumferential paths about the turbine axis. The seal member includes a sealing face that extends in a direction substantially parallel to the turbine axis. The seal member also includes a plurality of seal layers, wherein at least one of the seal layers includes at least one stress relief region for facilitating flexing of the first seal member.

  16. Disk cloning program 'Dolly+' for system management of PC Linux cluster

    International Nuclear Information System (INIS)

    Atsushi Manabe

    2001-01-01

    The Dolly+ is a Linux application program to clone files and disk partition image from a PC to many others. By using several techniques such as logical ring connection, multi threading and pipelining, it could achieve high performance and scalability. For example, in typical condition, installations to a hundred PCs takes almost equivalent time for two PCs. Together with the Intel PXE and the RedHat kickstart, automatic and very fast system installation and upgrading could be performed

  17. Development of an HIV-based cDNA expression cloning system.

    Science.gov (United States)

    van Maanen, Marc; Tidwell, Jennie K; Donehower, Lawrence A; Sutton, Richard E

    2003-07-01

    Expression cloning of cDNAs is a powerful tool with which to identify genes based on their specific functional properties. Here we describe the development of a cDNA library transfer system based on the human immunodeficiency virus type-1 (HIV). This system represents an improvement over current oncoretroviral cDNA expression systems in terms of target cell range and the inclusion of a selectable marker. By use of a simple packaging system, we were able to produce high-titer vector stocks from HIV vector-based cDNA libraries and demonstrate highly efficient cDNA expression cloning in three model experiments. First, HOS TK(-) cells, which are null for thymidine kinase (TK) expression, were transduced with an HIV-based cDNA library derived from primary human foreskin fibroblasts (HFFs) and functionally selected for TK expression. In a second experiment, hypoxanthine guanine phosphoribosyltransferase-1-deficient (HPRT(-)) fibroblasts were transduced with a T cell (PM1) line-derived cDNA library and selected for HPRT expression. Both TK (frequency 1 in 5.0 x 10(4)) and HPRT (frequency 1 in 2.0 x 10(4)) cDNAs were readily isolated from these HIV-based cDNA libraries. As a third example, we demonstrated the ability of this vector system to allow functional cDNA library screens to be performed in primary, mitotically inactive cell types. Using senescent HFFs as a target cell population, we were able to isolate SV40 large T antigen cDNA-containing clones (frequency 1 in 2.5 x 10(4)) based on their ability to overcome the senescence-induced block to cell proliferation. Thus, this system can be used to clone relatively low-abundance cDNAs based upon their expression. Because of the ability of HIV-based vectors to transduce primary and nondividing cells efficiently, this vector system will further broaden the range of cell types in which expression cloning studies can be performed.

  18. Biometric identification systems: the science of transaction facilitation

    Science.gov (United States)

    Rogers, Robert R.

    1994-10-01

    his fraud risk had been reduced to virtually zero. In highly sensitive security applications, such a system would be imperative -and when combined, if necessary, with other biometric identifiers such as signature and/or voice recognition for simultaneous verification, one would have a nearly foolproof system. These are the tools of what we call Transaction Facilitation, and this is the realm of Comparator Systems Corp. Our technological developments over the last ten years have moved our Company forward into a position of potential leadership in what is fast becoming a worldwide market, and it is toward this end that we have applied all of our efforts.

  19. Selection of chemotherapy for patient treatment utilizing a radiometric versus a cloning system

    Energy Technology Data Exchange (ETDEWEB)

    Von Hoff, D.D.; Forseth, B.J.; Turner, J.N.; Clark, G.M.; Warfel, L.E.

    1986-01-01

    From the 1950s to the 1970s, a number of in vitro systems that measured inhibition of glucose metabolism were used to predict the responsiveness of patients' tumors to chemotherapy. In vitro-in vivo correlations were excellent, with true positive predictions ranging from 68% to 96% and true negative predictions of 95% to 100%. The radiometric system is a new in vitro technique that measures the conversion of 14C-glucose to 14CO2. The system already has been utilized to screen prospective new antineoplastic agents for cytotoxicity. The present study was undertaken to determine if the radiometric system might be used to predict correctly the responsiveness of an individual patient's tumor to single-agent or combination-agent chemotherapy. Fifty-six tumor specimens were divided and tested for drug sensitivity in the radiometric system and a conventional human tumor clonning system. Overall, there was a significant correlation between in vitro and in vivo results for the conventional cloning system (P = 0.03). However, there was no significant relationship between in vitro and in vivo results for the radiometric system. The radiometric system consistently failed to predict the tumor's clinical sensitivity to single agents. A radiometric system is not useful in predicting the responsiveness of a patient's tumor to single agent chemotherapy and is not a replacement for the more biologically attractive human tumor cloning system.

  20. A high-throughput cloning system for reverse genetics in Trypanosoma cruzi.

    Science.gov (United States)

    Batista, Michel; Marchini, Fabricio K; Celedon, Paola A F; Fragoso, Stenio P; Probst, Christian M; Preti, Henrique; Ozaki, Luiz S; Buck, Gregory A; Goldenberg, Samuel; Krieger, Marco A

    2010-10-13

    The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches. We constructed a platform with a flexible structure allowing the exchange of various elements, such as promoters, fusion tags, intergenic regions or resistance markers. This platform is based on Gateway® technology, to ensure a fast and efficient cloning system. We obtained plasmid vectors carrying genes for fluorescent proteins (green, cyan or yellow), and sequences for the c-myc epitope, and tandem affinity purification or polyhistidine tags. The vectors were verified by successful subcellular localization of two previously characterized proteins (TcRab7 and PAR 2) and a putative centrin. For the tandem affinity purification tag, the purification of two protein complexes (ribosome and proteasome) was performed. We constructed plasmids with an efficient cloning system and suitable for use across various applications, such as protein localization and co-localization, protein partner identification and protein expression. This platform also allows vector customization, as the vectors were constructed to enable easy exchange of its elements. The development of this high

  1. A high-throughput cloning system for reverse genetics in Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Ozaki Luiz S

    2010-10-01

    Full Text Available Abstract Background The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches. Results We constructed a platform with a flexible structure allowing the exchange of various elements, such as promoters, fusion tags, intergenic regions or resistance markers. This platform is based on Gateway® technology, to ensure a fast and efficient cloning system. We obtained plasmid vectors carrying genes for fluorescent proteins (green, cyan or yellow, and sequences for the c-myc epitope, and tandem affinity purification or polyhistidine tags. The vectors were verified by successful subcellular localization of two previously characterized proteins (TcRab7 and PAR 2 and a putative centrin. For the tandem affinity purification tag, the purification of two protein complexes (ribosome and proteasome was performed. Conclusions We constructed plasmids with an efficient cloning system and suitable for use across various applications, such as protein localization and co-localization, protein partner identification and protein expression. This platform also allows vector customization, as the vectors were constructed to enable easy

  2. Seamless Ligation Cloning Extract (SLiCE) Cloning Method

    OpenAIRE

    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 D...

  3. Multipartite asymmetric quantum cloning

    International Nuclear Information System (INIS)

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

    2005-01-01

    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 A clones with fidelity F A and another set of M B clones with fidelity F B , the trade-off between these fidelities is analyzed, and particular cases of optimal N→M A +M B cloning machines are exhibited. We also present an optimal 1→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

  4. New analytical techniques to facilitate preformulation screening in propellant systems.

    Science.gov (United States)

    Mogalian, Erik; Kuehl, Philip J; Myrdal, Paul B

    2007-08-01

    The objective of these studies was to investigate the applicability of an online direct inject HPLC method for the preformulation screening of pharmaceutical agents in pressurized metered dose inhalers (MDIs). The technique was initially utilized for the solubility determination of solid solutes. This study explores the extension of the online direct inject method for the evaluation of drug stability in propellant systems as well as for the analysis of MDI vials crimped with metered valves. Through-life content analysis confirmed that a single vial may be repeatedly sampled, thus facilitating the stability evaluation of a single unit over time. The method was successfully used for evaluating the stability of a model drug, as a function of several different formulation configurations, with minimal sample numbers. Additionally, studies determined that after modifications were made to the injection coupler, the technique was also feasible for use with 50 and 100 microL metered valves, however further modifications are necessary for 25 microL valves.

  5. Cloning-Independent and Counterselectable Markerless Mutagenesis System in Streptococcus mutans▿

    Science.gov (United States)

    Xie, Zhoujie; Okinaga, Toshinori; Qi, Fengxia; Zhang, Zhijun; Merritt, Justin

    2011-01-01

    Insertion duplication mutagenesis and allelic replacement mutagenesis are among the most commonly utilized approaches for targeted mutagenesis in bacteria. However, both techniques are limited by a variety of factors that can complicate mutant phenotypic studies. To circumvent these limitations, multiple markerless mutagenesis techniques have been developed that utilize either temperature-sensitive plasmids or counterselectable suicide vectors containing both positive- and negative-selection markers. For many species, these techniques are not especially useful due to difficulties of cloning with Escherichia coli and/or a lack of functional negative-selection markers. In this study, we describe the development of a novel approach for the creation of markerless mutations. This system employs a cloning-independent methodology and should be easily adaptable to a wide array of Gram-positive and Gram-negative bacterial species. The entire process of creating both the counterselection cassette and mutation constructs can be completed using overlapping PCR protocols, which allows extremely quick assembly and eliminates the requirement for either temperature-sensitive replicons or suicide vectors. As a proof of principle, we used Streptococcus mutans reference strain UA159 to create markerless in-frame deletions of 3 separate bacteriocin genes as well as triple mutants containing all 3 deletions. Using a panel of 5 separate wild-type S. mutans strains, we further demonstrated that the procedure is nearly 100% efficient at generating clones with the desired markerless mutation, which is a considerable improvement in yield compared to existing approaches. PMID:21948849

  6. Cloning-independent and counterselectable markerless mutagenesis system in Streptococcus mutans.

    Science.gov (United States)

    Xie, Zhoujie; Okinaga, Toshinori; Qi, Fengxia; Zhang, Zhijun; Merritt, Justin

    2011-11-01

    Insertion duplication mutagenesis and allelic replacement mutagenesis are among the most commonly utilized approaches for targeted mutagenesis in bacteria. However, both techniques are limited by a variety of factors that can complicate mutant phenotypic studies. To circumvent these limitations, multiple markerless mutagenesis techniques have been developed that utilize either temperature-sensitive plasmids or counterselectable suicide vectors containing both positive- and negative-selection markers. For many species, these techniques are not especially useful due to difficulties of cloning with Escherichia coli and/or a lack of functional negative-selection markers. In this study, we describe the development of a novel approach for the creation of markerless mutations. This system employs a cloning-independent methodology and should be easily adaptable to a wide array of Gram-positive and Gram-negative bacterial species. The entire process of creating both the counterselection cassette and mutation constructs can be completed using overlapping PCR protocols, which allows extremely quick assembly and eliminates the requirement for either temperature-sensitive replicons or suicide vectors. As a proof of principle, we used Streptococcus mutans reference strain UA159 to create markerless in-frame deletions of 3 separate bacteriocin genes as well as triple mutants containing all 3 deletions. Using a panel of 5 separate wild-type S. mutans strains, we further demonstrated that the procedure is nearly 100% efficient at generating clones with the desired markerless mutation, which is a considerable improvement in yield compared to existing approaches.

  7. Cloned cattle derived from a novel zona-free embryo reconstruction system

    NARCIS (Netherlands)

    Oback, B; Wiersema, AT; Gaynor, P; Laible, G; Tucker, FC; Oliver, JE; Miller, AL; Troskie, HE; Wilson, KL; Forsyth, JT; Berg, MC; Cockrem, K; Mcmillan, [No Value; Tervit, HR; Wells, DN

    2003-01-01

    As the demand for cloned embryos and offspring increases, the need arises for the development of nuclear transfer procedures that are improved in both efficiency and ease of operation. Here, we describe a novel zona-free cloning method that doubles the throughput in cloned bovine embryo production

  8. What is Cloning?

    Science.gov (United States)

    Donate Home Cloning What is Cloning What is Cloning Clones are organisms that are exact genetic copies. ... clones made through modern cloning technologies. How Is Cloning Done? Many people first heard of cloning when ...

  9. Cloning and expression of the HIV protein in Escherichia coli cell-free system.

    Science.gov (United States)

    Chen, Haiqin; Xu, Zhinan; Yin, Xiufei; Cen, Peilin

    2007-11-01

    Sixteen kinds of human immunodeficiency virus (HIV) target genes were cloned by polymerase chain reaction (PCR) amplification, and specific plasmids were constructed as the templates for the expression of these genes in the cell-free system. Similarly, the linear PCR templates of these genes for cell-free protein expression were also constructed by using two PCR amplification process. These different templates can be employed to biosynthesize HIV proteins in the cell-free system simultaneously and can be adapted for some high-throughput processes. HIV protease (P10) was performed as a target protein, and two different templates (plasmid and PCR product) were prepared and used for P10 expression in the Escherichia coli cell-free system. The target protein P10 was detected in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels either by using a plasmid template or by a PCR template. These results are promising and helpful to develop a high throughput process for drug discovery.

  10. Cloning Mice.

    Science.gov (United States)

    Ogura, Atsuo

    2017-08-01

    Viable and fertile mice can be generated by somatic nuclear transfer into enucleated oocytes, presumably because the transplanted somatic cell genome becomes reprogrammed by factors in the oocyte. The first somatic cloned offspring of mice were obtained by directly injecting donor nuclei into recipient enucleated oocytes. When this method is used (the so-called Honolulu method of somatic cell nuclear transfer [SCNT]), the donor nuclei readily and completely condense within the enucleated metaphase II-arrested oocytes, which contain high levels of M-phase-promoting factor (MPF). It is believed that the condensation of the donor chromosomes promotes complete reprogramming of the donor genome within the mouse oocytes. Another key to the success of mouse cloning is the use of blunt micropipettes attached to a piezo impact-driving micromanipulation device. This system saves a significant amount of time during the micromanipulation of oocytes and thus minimizes the loss of oocyte viability in vitro. For example, a group of 20 oocytes can be enucleated within 10 min by an experienced operator. This protocol is composed of seven parts: (1) preparing micropipettes, (2) setting up the enucleation and injection micropipettes, (3) collecting and enucleating oocytes, (4) preparing nucleus donor cells, (5) injecting donor nuclei, (6) activating embryos and culturing, and (7) transferring cloned embryos. © 2017 Cold Spring Harbor Laboratory Press.

  11. Cloning and Study of Expression of Helicobacter Pylori FlaAGene in Eukaryotic System

    Directory of Open Access Journals (Sweden)

    Motaram Sadeghi

    2017-07-01

    Full Text Available Background: Helicobacter pylori is the most common bacterium causing chronic infections worldwide.Expression of flagella and bacterial motility are very important incolonization and virulence. FlaA geneis one of theflagellin-encoding genes that play the key role in the colonization and bacterial motility and it has a significant impact in immunization. The aim of this study wasto design, construction and the evaluation of the Helicobacter pylori flaA gene expression in eukaryotic cells. MaterialandMethods: In this experimental study, genomic DNA was purified from the Helicobacter pylori standard strain and flaA gene was amplified and isolated by PCR method with use of the specific primers. Then, this gene was cloned into pTZ vector by T/A cloning technique. In order to flaA gene expression and generation of final construct, the flaA gene was removed from pTZ plasmid and sub-cloned into the pcDNA3.1 (- expression vector. The pcDNA3.1 (--flaA construct was transformed into CHO cells by electroporation, and flaA eukaryotic gene expression was studied on SDS-PAGE. Results: The results showed that flaA gene PCR product was cloned into pTZ vector and amplified in Escherichia coli TOP10F strain. Also the enzymatic digestion and sequencing showed that the pcDNA3.1 (--flaA was performed. Finally, the evaluation of the Helicobacter pyloriflaA gene expression in CHO cells showed that the generated gene construct can expressed the flaA product in eukaryotic system, successfully. Conclusion: Given that the pcDNA3.1 (--flaA as a final construct is able to express the flaA protein of the Helicobacterpylori in animal cells. Flagellin protein is one of the important antigens of the bacteriumSo we can properly say that gene pcDNA3.1(--flaA consteuct is an appropriate candidate for use in the field of gene vaccines against Helicobacter pylori and in future researches can be used to check the immunization in laboratory animals. 

  12. An integrative analysis of reprogramming in human isogenic system identified a clone selection criterion.

    Science.gov (United States)

    Shutova, Maria V; Surdina, Anastasia V; Ischenko, Dmitry S; Naumov, Vladimir A; Bogomazova, Alexandra N; Vassina, Ekaterina M; Alekseev, Dmitry G; Lagarkova, Maria A; Kiselev, Sergey L

    2016-01-01

    The pluripotency of newly developed human induced pluripotent stem cells (iPSCs) is usually characterized by physiological parameters; i.e., by their ability to maintain the undifferentiated state and to differentiate into derivatives of the 3 germ layers. Nevertheless, a molecular comparison of physiologically normal iPSCs to the "gold standard" of pluripotency, embryonic stem cells (ESCs), often reveals a set of genes with different expression and/or methylation patterns in iPSCs and ESCs. To evaluate the contribution of the reprogramming process, parental cell type, and fortuity in the signature of human iPSCs, we developed a complete isogenic reprogramming system. We performed a genome-wide comparison of the transcriptome and the methylome of human isogenic ESCs, 3 types of ESC-derived somatic cells (fibroblasts, retinal pigment epithelium and neural cells), and 3 pairs of iPSC lines derived from these somatic cells. Our analysis revealed a high input of stochasticity in the iPSC signature that does not retain specific traces of the parental cell type and reprogramming process. We showed that 5 iPSC clones are sufficient to find with 95% confidence at least one iPSC clone indistinguishable from their hypothetical isogenic ESC line. Additionally, on the basis of a small set of genes that are characteristic of all iPSC lines and isogenic ESCs, we formulated an approach of "the best iPSC line" selection and confirmed it on an independent dataset.

  13. Viabilidade econômica de diferentes sistemas de sangria em clones de seringueira Economic viability of different tapping systems in rubber tree clones

    Directory of Open Access Journals (Sweden)

    Juliano Quarteroli Silva

    2007-03-01

    Full Text Available O objetivo deste trabalho foi avaliar o desempenho produtivo e os aspectos econômicos de três clones de seringueira [Hevea brasiliensis (Willd. ex Adr. de Juss. Muell. Arg.], sob nove sistemas de sangria. O experimento foi instalado sob delineamento de blocos ao acaso com parcelas subdivididas no tempo. Os tratamentos principais foram os clones PR 255, RRIM 600 e GT 1, submetidos aos seguintes sistemas de sangria: ½S d/3 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/3 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/4 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/4 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/5 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/5 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/7 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/7 6d/7.11m/y.ET 5,0% Pa La 8/y e ½S d/2 6d/7.11m/y (testemunha. As variáveis estudadas foram: perímetro do caule, produtividade de borracha seca e secamento do painel. Também foi avaliada a viabilidade econômica dos sistemas de sangria. Observaram-se maior produtividade e rentabilidade dos sistemas ½S d/3.ET 2,5% 8/y para os clones PR 255 e RRIM 600 e ½S d/7.ET 2,5% 8/y para o clone GT 1, comparados com a testemunha. A maior e a menor porcentagem de secamento do painel foram observadas nos sistemas ½S d/3 ET 5,0% 8/y e ½S d/7.ET 5,0% 8/y, respectivamente.The objective of this work was to evaluate yield performance and economic aspects of three clones of rubber tree [Hevea brasiliensis (Willd. ex Adr. de Juss. Muell. Arg.], in different tapping systems. The trial was placed under randomized blocks design with split-plot in time. The main treatments were PR 255, RRIM 600 and GT 1 clones submitted to nine systems of tapping: ½S d/3 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/3 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/4 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/4 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/5 6d/7.11m/y.ET 2,5% Pa La 8/y; ½S d/5 6d/7.11m/y.ET 5,0% Pa La 8/y; ½S d/7 6d/7.11m/y.ET 2,5% Pa La 8/y ; ½S d/7 6d/7.11m/y.ET 5,0% Pa La 8/y and ½S d/2 6d/7.11m/y (check. The

  14. The BsaHI restriction-modification system: Cloning, sequencing and analysis of conserved motifs

    Directory of Open Access Journals (Sweden)

    Roberts Richard J

    2008-05-01

    Full Text Available Abstract Background Restriction and modification enzymes typically recognise short DNA sequences of between two and eight bases in length. Understanding the mechanism of this recognition represents a significant challenge that we begin to address for the BsaHI restriction-modification system, which recognises the six base sequence GRCGYC. Results The DNA sequences of the genes for the BsaHI methyltransferase, bsaHIM, and restriction endonuclease, bsaHIR, have been determined (GenBank accession #EU386360, cloned and expressed in E. coli. Both the restriction endonuclease and methyltransferase enzymes share significant similarity with a group of 6 other enzymes comprising the restriction-modification systems HgiDI and HgiGI and the putative HindVP, NlaCORFDP, NpuORFC228P and SplZORFNP restriction-modification systems. A sequence alignment of these homologues shows that their amino acid sequences are largely conserved and highlights several motifs of interest. We target one such conserved motif, reading SPERRFD, at the C-terminal end of the bsaHIR gene. A mutational analysis of these amino acids indicates that the motif is crucial for enzymatic activity. Sequence alignment of the methyltransferase gene reveals a short motif within the target recognition domain that is conserved among enzymes recognising the same sequences. Thus, this motif may be used as a diagnostic tool to define the recognition sequences of the cytosine C5 methyltransferases. Conclusion We have cloned and sequenced the BsaHI restriction and modification enzymes. We have identified a region of the R. BsaHI enzyme that is crucial for its activity. Analysis of the amino acid sequence of the BsaHI methyltransferase enzyme led us to propose two new motifs that can be used in the diagnosis of the recognition sequence of the cytosine C5-methyltransferases.

  15. Expert Systems as a Mindtool To Facilitate Mental Model Learning.

    Science.gov (United States)

    Mason-Mason, Susan Dale; Tessmer, Martin A.

    2000-01-01

    This exploratory study investigated whether the process of constructing an expert system model promotes the formation of expert-like mental models. Discusses expert systems as mindtools, expert systems as learning tools, the assessment of mental models, results of pretests and posttests, and future research. (Contains 56 references.) (Author/LRW)

  16. Molecular cloning and analysis of the ergopeptine assembly system in the ergot fungus Claviceps purpurea.

    Science.gov (United States)

    Correia, Telmo; Grammel, Nicolas; Ortel, Ingo; Keller, Ullrich; Tudzynski, Paul

    2003-12-01

    Claviceps purpurea produces the pharmacological important ergopeptines, a class of cyclol-structured alkaloid peptides containing D-lysergic acid. These compounds are assembled from D-lysergic acid and three different amino acids by the nonribosomal peptide synthetase enzymes LPS1 and LPS2. Cloning of alkaloid biosynthesis genes from C. purpurea has revealed a gene cluster including two NRPS genes, cpps 1 and cpps 2. Protein sequence data had assigned earlier cpps1 to encode the trimodular LPS1 assembling the tripeptide portion of ergopeptines. Here, we show by transcriptional analysis, targeted inactivation, analysis of disruption mutants, and heterologous expression that cpps 2 encodes the monomodular LPS2 responsible for D-lysergic acid activation and incorporation into the ergopeptine backbone. The presence of two distinct NRPS subunits catalyzing formation of ergot peptides is the first example of a fungal NRPS system consisting of different NRPS subunits.

  17. TGATG vector: a new expression system for cloned foreign genes in Escherichia coli cells.

    Science.gov (United States)

    Mashko, S V; Veiko, V P; Lapidus, A L; Lebedeva, M I; Mochulsky, A V; Shechter, I I; Trukhan, M E; Ratmanova, K I; Rebentish, B A; Kaluzhsky, V E

    1990-03-30

    A TGATG vector system was developed that allows for the construction of hybrid operons with partially overlapping genes, employing the effects of translational coupling to optimize expression of cloned cistrons in Escherichia coli. In this vector system (plasmid pPR-TGATG-1), the coding region of a foreign gene is attached to the ATG codon situated on the vector, to form the hybrid operon transcribed from the phage lambda PR promoter. The cloned gene is the distal cistron of this hybrid operon ('overlappon'). The efficiently translated cro'-cat'-'trpE hybrid cistron is proximal to the promoter. The coding region of this artificial fused cistron [the length of the corresponding open reading frame is about 120 amino acids (aa)] includes the following: the N-terminal portions of phage lambda Cro protein (20 aa), the CAT protein of E. coli (72 aa) and 3' C-terminal codons of the E. coli trpE gene product. At the 3'-end of the cro'-cat'-'trpE fused cistron there is a region for efficient translation reinitiation: a Shine-Dalgarno sequence of the E. coli trpD gene and the overlapping stop and start codons (TGATG). In this sequence, the last G is the first nucleotide of the unique SacI-recognition site (GAGCT decreases C) and so integration of the structural part of the foreign gene into the vector plasmid may be performed using blunt-end DNA linking after the treatment of pPR-TGATG-1 with SacI and E. coli DNA polymerase I or its Klenow fragment.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Gateway Recombinational Cloning.

    Science.gov (United States)

    Reece-Hoyes, John S; Walhout, Albertha J M

    2018-01-02

    The Gateway recombinatorial cloning system was developed for cloning multiple DNA fragments in parallel (e.g., in 96-well formats) in a standardized manner using the same enzymes. Gateway cloning is based on the highly specific integration and excision reactions of bacteriophage λ into and out of the Escherichia coli genome. Because the sites of recombination (" att " sites) are much longer (25-242 bp) than restriction sites, they are extremely unlikely to occur by chance in DNA fragments. Therefore, the same recombination enzyme can be used to robustly clone many different fragments of variable size in parallel reactions. © 2018 Cold Spring Harbor Laboratory Press.

  19. Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system.

    Science.gov (United States)

    Nakamura, Tsuyoshi; Omasa, Takeshi

    2015-09-01

    Therapeutic antibodies are commonly produced by high-expressing, clonal and recombinant Chinese hamster ovary (CHO) cell lines. Currently, CHO cells dominate as a commercial production host because of their ease of use, established regulatory track record, and safety profile. CHO-K1SV is a suspension, protein-free-adapted CHO-K1-derived cell line employing the glutamine synthetase (GS) gene expression system (GS-CHO expression system). The selection of high-producing mammalian cell lines is a crucial step in process development for the production of therapeutic antibodies. In general, cloning by the limiting dilution method is used to isolate high-producing monoclonal CHO cells. However, the limiting dilution method is time consuming and has a low probability of monoclonality. To minimize the duration and increase the probability of obtaining high-producing clones with high monoclonality, an automated single cell-based clone selector, the ClonePix FL system, is available. In this study, we applied the high-throughput ClonePix FL system for cell line development using CHO-K1SV cells and investigated efficient conditions for single cell-based clone selection. CHO-K1SV cell growth at the pre-picking stage was improved by optimizing the formulation of semi-solid medium. The efficiency of picking and cell growth at the post-picking stage was improved by optimization of the plating time without decreasing the diversity of clones. The conditions for selection, including the medium formulation, were the most important factors for the single cell-based clone selection system to construct a high-producing CHO cell line. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  20. Growth of single T cells and single thymocytes in a high cloning efficiency filler-cell free microculture system.

    Science.gov (United States)

    Chen, W F; Ewing, T; Scollay, R; Shortman, K

    1988-01-01

    A high cloning-efficiency microculture system is described in which single T cells, stimulated to divide by phorbol ester and calcium ionophore, grow rapidly under the influence of purified growth factors in the absence of other cells. The kinetics of clonal growth has been monitored over a five day period by phase-contrast microscopy. Mature peripheral T cells, and mature subpopulations from the thymus, responded with a cloning efficiency over 80%; they required IL-2 as a minimum but several other factors enhanced growth. Ly2+L3T4- thymocytes (mean doubling time 10.4 hr) grew more rapidly than Ly2-L3T4+ thymocytes (mean doubling time 15.2 hr). Early (Ly2-L3T4-) thymocytes responded with a cloning efficiency of 60%; their efficient growth was dependent on both IL-1 and IL-2. The typical Ly2+L3T4+ cortical thymocyte did not grow under these conditions.

  1. Cloning of a two-component signal transduction system of Xanthomonas campestris pv. phaseoli var. fuscans strain BXPF65

    DEFF Research Database (Denmark)

    Chan, JWYF; Maynard, Scott; Goodwin, PH

    1998-01-01

    A putative two-component signal transduction system was amplified and cloned from the plant pathogenic bacterium Xanthomonas campestris pv. phaseoli var. fuscans isolate BXPF65. The 620 bp amplified fragment was sequenced and analyzed with the BLAST Enhanced Alignment Utility (BEAUTY). BEAUTY ana...

  2. IDENTIFICATION OF ACTIVE BACTERIAL COMMUNITIES IN A MODEL DRINKING WATER BIOFILM SYSTEM USING 16S RRNA-BASED CLONE LIBRARIES

    Science.gov (United States)

    Recent phylogenetic studies have used DNA as the target molecule for the development of environmental 16S rDNA clone libraries. As DNA may persist in the environment, DNA-based libraries cannot be used to identify metabolically active bacteria in water systems. In this study, a...

  3. A Recommendation System to Facilitate Business Process Modeling.

    Science.gov (United States)

    Deng, Shuiguang; Wang, Dongjing; Li, Ying; Cao, Bin; Yin, Jianwei; Wu, Zhaohui; Zhou, Mengchu

    2017-06-01

    This paper presents a system that utilizes process recommendation technology to help design new business processes from scratch in an efficient and accurate way. The proposed system consists of two phases: 1) offline mining and 2) online recommendation. At the first phase, it mines relations among activity nodes from existing processes in repository, and then stores the extracted relations as patterns in a database. At the second phase, it compares the new process under construction with the premined patterns, and recommends proper activity nodes of the most matching patterns to help build a new process. Specifically, there are three different online recommendation strategies in this system. Experiments on both real and synthetic datasets are conducted to compare the proposed approaches with the other state-of-the-art ones, and the results show that the proposed approaches outperform them in terms of accuracy and efficiency.

  4. Automated cloning methods.; TOPICAL

    International Nuclear Information System (INIS)

    Collart, F.

    2001-01-01

    Argonne has developed a series of automated protocols to generate bacterial expression clones by using a robotic system designed to be used in procedures associated with molecular biology. The system provides plate storage, temperature control from 4 to 37 C at various locations, and Biomek and Multimek pipetting stations. The automated system consists of a robot that transports sources from the active station on the automation system. Protocols for the automated generation of bacterial expression clones can be grouped into three categories (Figure 1). Fragment generation protocols are initiated on day one of the expression cloning procedure and encompass those protocols involved in generating purified coding region (PCR)

  5. Breeding of transgenic cattle for human coagulation factor IX by a combination of lentiviral system and cloning.

    Science.gov (United States)

    Monzani, P S; Sangalli, J R; De Bem, T H C; Bressan, F F; Fantinato-Neto, P; Pimentel, J R V; Birgel-Junior, E H; Fontes, A M; Covas, D T; Meirelles, F V

    2013-02-28

    Recombinant coagulation factor IX must be produced in mammalian cells because FIX synthesis involves translational modifications. Human cell culture-based expression of human coagulation factor IX (hFIX) is expensive, and large-scale production capacity is limited. Transgenic animals may greatly increase the yield of therapeutic proteins and reduce costs. In this study, we used a lentiviral system to obtain transgenic cells and somatic cell nuclear transfer (SCNT) to produce transgenic animals. Lentiviral vectors carrying hFIX driven by 3 bovine β-casein promoters were constructed. Bovine epithelial mammary cells were transduced by lentivirus, selected with blasticidin, plated on extracellular matrix, and induced by lactogenic hormones; promoter activity was evaluated by quantitative PCR. Transcriptional activity of the 5.335-kb promoter was 6-fold higher than the 3.392- and 4.279-kb promoters, which did not significantly differ. Transgenic bovine fibroblasts were transduced with lentivirus carrying the 5.335-kb promoter and used as donor cells for SCNT. Cloned transgenic embryo production yielded development rates of 28.4%, similar to previous reports on cloned non-transgenic embryos. The embryos were transferred to recipient cows (N = 21) and 2 births of cloned transgenic cattle were obtained. These results suggest combination of the lentiviral system and cloning may be a good strategy for production of transgenic cattle.

  6. The genus Amycolatopsis: Indigenous plasmids, cloning vectors and gene transfer systems.

    Science.gov (United States)

    Malhotra, S; Lal, R

    2007-03-01

    The genus Amycolatopsis is a member of the phylogenetic group nocardioform actinomycetes. Most of the members of the genus Amycolatopsis are known to produce antibiotics. Additionally, members of this genus have been reported to metabolize aromatic compounds as the sole sources of carbon and energy. Development of genetic manipulation in Amycolatopsis has progressed slowly due to paucity of genetic tools and methods. The occurrence of indigenous plasmids in different species of Amycolatopsis is not very common. Till date, only three indigenous plasmids viz., pMEA100, pMEA300 and pA387 have been reported in Amycolatopsis species. Various vectors based on the indigenous plasmids, pMEA100, pMEA300 and pA387, have been constructed. These vectors have proved useful for molecular genetics studies of actinomycetes. Molecular genetic work with Amycolatopsis strains is not easy, since transformation methods have to be developed, or at least optimized, for each particular strain. Nonetheless, methods for efficient transformation (polyethyleneglycol (PEG) induced protoplast transformation, transformation by electroporation and direct transformation) have been developed and used successfully for the introduction of DNA into several Amycolatopsis species. The construction of plasmid cloning vectors and the development of gene transfer systems has opened up possibilities for studying the molecular genetics of these bacteria.

  7. The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs

    Directory of Open Access Journals (Sweden)

    Masahiro Sato

    2016-08-01

    Full Text Available The introduction of multigene constructs into single cells is important for improving the performance of domestic animals, as well as understanding basic biological processes. In particular, multigene constructs allow the engineering and integration of multiple genes related to xenotransplantation into the porcine genome. The piggyBac (PB transposon system allows multiple genes to be stably integrated into target genomes through a single transfection event. However, to our knowledge, no attempt to introduce multiple genes into a porcine genome has been made using this system. In this study, we simultaneously introduced seven transposons into a single porcine embryonic fibroblast (PEF. PEFs were transfected with seven transposons containing genes for five drug resistance proteins and two (red and green fluorescent proteins, together with a PB transposase expression vector, pTrans (experimental group. The above seven transposons (without pTrans were transfected concomitantly (control group. Selection of these transfected cells in the presence of multiple selection drugs resulted in the survival of several clones derived from the experimental group, but not from the control. PCR analysis demonstrated that approximately 90% (12/13 tested of the surviving clones possessed all of the introduced transposons. Splinkerette PCR demonstrated that the transposons were inserted through the TTAA target sites of PB. Somatic cell nuclear transfer (SCNT using a PEF clone with multigene constructs demonstrated successful production of cloned blastocysts expressing both red and green fluorescence. These results indicate the feasibility of this PB-mediated method for simultaneous transfer of multigene constructs into the porcine cell genome, which is useful for production of cloned transgenic pigs expressing multiple transgenes.

  8. Facilitating Constructive Alignment in Power Systems Engineering Education Using Free and Open-Source Software

    Science.gov (United States)

    Vanfretti, L.; Milano, F.

    2012-01-01

    This paper describes how the use of free and open-source software (FOSS) can facilitate the application of constructive alignment theory in power systems engineering education by enabling the deep learning approach in power system analysis courses. With this aim, this paper describes the authors' approach in using the Power System Analysis Toolbox…

  9. A Systemic-Constructivist Approach to the Facilitation and Debriefing of Simulations and Games

    Science.gov (United States)

    Kriz, Willy Christian

    2010-01-01

    This article introduces some basic concepts of a systemic-constructivist perspective. These show that gaming simulation corresponds closely to a systemic-constructivist approach to learning and instruction. Some quality aspects of facilitating and debriefing simulation games are described from a systemic-constructivist point of view. Finally, a…

  10. Agronomical performance and profitability of exploitation systems in four rubber tree clones in São Paulo state

    Directory of Open Access Journals (Sweden)

    Juliano Quarteroli Silva

    2010-12-01

    Full Text Available The exploitation or tapping of the rubber tree, Hevea brasiliensis (Willd. ex Adr. de Juss. Muell. Arg. is one of the most important cultural practices in determining useful life, yield and accounts for a major part of the total production costs in rubber farming. The objective of this work was to evaluate yield performance and economic aspects of rubber tree clones submitted to diverse tapping systems. The trial was placed in Guararapes city, São Paulo State, Brazil, in a randomized block design with split-plot in time. The plots consisted of the IAN 873, PR 261, RRI M 600 and RRI M 701 clones. The tapping systems consisted the subplots, where: ½S = tapping of half spiral cut; d/2, d/3, d/4, d/5 and d/7 = tapping every 2, 3, 4, 5 and 7 days, respectively; 11 m/y = tapping during eleven months per year; ET = ethephon (stimulant; Pa = panel application; La = lace application; 8/y = eight applications per year. The five experimental years were the sub-subplots and the ½S d/2 system was used as control. The analyzed variables were girth, dry rubber yield, tapping panel dryness and economic profitability. The ½S d/3 ET 2.5% and ½S d/4 ET 2.5% tapping systems provide the highest yield and profitability per hectare per year for the RRI M 600 and PR 261 clones. For the IAN 873 and RRI M 701 clones the yield superiority occurs in high tapping frequency; however the best profitability is obtained in the ½S d/7.ET 2.5% system.

  11. Functional cloning of Vibrio parahaemolyticus type III secretion system 1 in Escherichia coli K-12 strain as a molecular syringe.

    Science.gov (United States)

    Akeda, Yukihiro; Kimura, Tomomi; Yamasaki, Aiko; Kodama, Toshio; Iida, Tetsuya; Honda, Takeshi; Oishi, Kazunori

    2012-10-19

    The type III secretion system (T3SS) of gram-negative bacteria involves dedicated protein translocation machinery that directly injects proteins into target cells. Pathogenic bacteria already benefit from this unique system. The successful functional cloning of this useful tool into non-pathogenic bacteria would help establish novel clinical and basic biotechnology strategies in areas such as vaccine administration, the development of screening systems for anti-T3SS drugs and the target-specific delivery of bioactive compounds. In this study, we successfully cloned the Vibrio parahaemolyticus T3SS1 genetic locus into a non-pathogenic Escherichia coli K-12 strain. Assays performed here revealed that the T3SS1 cloned into the E. coli K-12 strain has the ability to translocate V. parahaemolyticus T3SS1 secreted proteins. Importantly, we also observed this system to allow the E. coli K-12 strain to inject foreign protein, as well as the V. parahaemolyticus T3SS effector, into cultured cells. These results demonstrate a prospective useful tool with experimental and therapeutic applications. Copyright © 2012 Elsevier Inc. All rights reserved.

  12. Discovery of Functional Toxin/Antitoxin Systems in Bacteria by Shotgun Cloning

    Energy Technology Data Exchange (ETDEWEB)

    Sberro, Hila; Leavitt, Azita; Kiro, Ruth; Koh, Eugene; Peleg, Yoav; Qimron, Udi; Sorek, Rotem

    2013-04-01

    Toxin-antitoxin (TA) modules, composed of a toxic protein and a counteracting antitoxin, play important roles in bacterial physiology. We examined the experimental insertion of 1.5 million genes from 388 microbial genomes into an Escherichia coli host using over 8.5 million random clones. This revealed hundreds of genes (toxins) that could only be cloned when the neighboring gene (antitoxin) was present on the same clone. Clustering of these genes revealed TA families widespread in bacterial genomes, some of which deviate from the classical characteristics previously described for such modules. Introduction of these genes into E. coli validated that the toxin toxicity is mitigated by the antitoxin. Infection experiments with T7 phage showed that two of the new modules can provide resistance against phage. Moreover, our experiments revealed an 'anti-defense' protein in phage T7 that neutralizes phage resistance. Our results expose active fronts in the arms race between bacteria and phage.

  13. A yeast-based model system for cloning secreted and membrane proteins

    Directory of Open Access Journals (Sweden)

    MARCELO J. SURPILI

    2002-12-01

    Full Text Available The targeting of proteins to cell organelles and membranes, or of proteins destined to secretion, is coordinated by signal sequences located at the 5´-end of their respective genes. A signal sequence trap system was envisaged in which a truncated version of the yeast acid phosphatase pho5 gene lacking the start codon and signal sequence could serve as a reporter gene. A fraction enriched in 5´-end fragments obtained by PCR from a potato guard-cell cDNA library was cloned in frame to the acid phosphatase gene and the acid phosphatase activity was assayed directly in yeast colonies grown on selective medium. Putative signal sequences targeting the acid phosphatase to the membrane or to the outside of the cell were used to screen the cDNA bank in order to recover the original full-size sequence which gave rise to the signal sequence. Two unknown sequences displaying marked tissue-specific expression were retrieved, one of them (YE139 with a higher expression level in green buds and stem cells, and the other one (YE290 with a higher expression level in androceum, gyneceum, and roots. The limitations of the system are further analyzed using other sequences as control.O direcionamento de proteínas a organelas e à membrana celular, ou de proteínas a serem secretadas, é coordenado por seqüências sinalizadoras localizadas na extremidade 5´ de seus respectivos genes que codificam peptídeos-sinal. Este trabalho analisa um sistema para seleção de seqüências sinalizadoras utilizando uma fosfatase ácida de levedura, enzima reconhecidamente secretada por estes organismos, desprovida de seu códon de iniciação e de sua seqüência sinalizadora, como gene repórter. Uma fração enriquecida em fragmentos provenientes da região 5´ de uma biblioteca de cDNA de células-guarda de batata foi inserida in frame ao gene truncado da fosfatase ácida em vetores apropriados. Após a transformação em leveduras, a atividade da fosfatase ácida foi

  14. Hand-made cloned goat (Capra hircus) embryos—a comparison of different donor cells and culture systems.

    Science.gov (United States)

    Akshey, Yogesh S; Malakar, Dhruba; De, Arun K; Jena, Manoj K; Garg, Shweta; Dutta, Rahul; Pawar, Sachin Kumar; Mukesh, Manisha

    2010-10-01

    Nuclear transfer is a very effective method for propagation of valuable, extinct, and endangered animals. Hand-made cloning (HMC) is an efficient alternative to the conventional micromanipulator-based technique in some domestic species. The present study was carried out for the selection of suitable somatic cells as a nuclear donor and development of an optimum culture system for in vitro culture of zona-free goat cloned embryos. Cleavage and blastocyst rates were observed 72.06 ± 2.94% and 0% for fresh cumulus cells, 81.95 ± 3.40% and 12.74 ± 2.12% for cultured cumulus cells, and 92.94 ± 0.91% and 23.78 ± 3.33% for fetal fibroblast cells, respectively. There was a significant (p WOW) (15.84 ± 2.12 %) and microdrops (MD) (0.7 ± 0.7%). Furthermore, cleavage and blastocyst production rates were significantly (p WOW (15.84 ± 2.12%) than the MD (0.7 ± 0.7%) system. The quality of HMC blastocysts was studied by differential staining. Genetic similarity was confirmed by polymerase chain reaction (PCR)-based amplification of the second exon of the MHC class II DRB gene, which gave similar bands in electrophoresis (286 bp) both in cloned embryos and donor cells. In conclusion, the present study describes that the fetal fibroblast cell is a suitable candidate as nuclear donor, and the flat surface culture system is suitable for zona-free blastocyst development by the hand-made cloning technique in the goat.

  15. Sistemas de explotação de seringueira utilizados em clones asiáticos Prang Besar no Oeste paulista Rubber tree exploitation systems utilized in asian clones Prang Besar, in the West of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Juliano Quarteroli Silva

    2007-07-01

    Full Text Available O objetivo deste trabalho foi avaliar o desempenho produtivo e os aspectos fisiológicos e econômicos de quatro clones asiáticos Prang Besar de seringueira [Hevea brasiliensis (Willd. ex A. Juss. Muell. Arg.], sob diferentes sistemas de sangria. O experimento foi instalado na Fazenda Santa Gilda, Município de Guararapes, SP, em delineamento de blocos ao acaso, com parcelas subdivididas no tempo. Os tratamentos principais foram os clones PB 260, PB 235, PB 330 e PB 217, submetidos a nove sistemas de sangria: ½S d/3.ET 2,5% 8/y; ½S d/3.ET 5% 8/y; ½S d/4.ET 2,5% 8/y; ½S d/4.ET 5% 8/y; ½S d/5.ET 2,5% 8/y; ½S d/5.ET 5% 8/y; ½S d/7.ET 2,5% 8/y; ½S d/7.ET 5% 8/y e ½S d/2 (testemunha. As variáveis avaliadas foram: perímetro do caule, produção de borracha seca, secamento do painel; foi avaliada, também, a viabilidade econômica. Os resultados mostram a superioridade econômica dos sistemas ½S d/5.ET 2,5% 8/y e ½S d/7.ET 5% 8/y, para o clone PB 260; ½S d/7.ET 2,5% 8/y para o clone PB 235; e ½S d/3.ET 2,5% 8/y e ½S d/3.ET 5% 8/y, para os clones PB 330 e PB 217, comparados com a testemunha. A menor incidência de secamento do painel foi observada no sistema ½S d/7.ET 5% 8/y, exceto para o clone PB 235.The objective of this work was to evaluate the yield performance, and economic and physiologic aspects of four Asian clones Prang Besar of rubber tree [Hevea brasiliensis (Willd. ex A. Juss. Muell. Arg.], in different tapping systems. The trial was placed at Santa Gilda farm, City of Guararapes, São Paulo State, Brazil, in a randomized block design with split-plot in time. The main treatments were PB 260, PB 235, PB 330 and PB 217 clones submitted to nine systems of tapping: ½S d/3.ET 2.5% 8/y; ½S d/3.ET 5% 8/y; ½S d/4.ET 2.5% 8/y; ½S d/4.ET 5% 8/y; ½S d/5.ET 2.5% 8/y; ½S d/5.ET 5% 8/y; ½S d/7.ET 2.5% 8/y; ½S d/7.ET 5% 8/y and ½S d/2 (check. The variables evaluated were: girth increment, dry rubber production, brown bast; it

  16. In vitro multiplication of Colocasia esculenta clone ‘INIVIT MC-2001’ in semi-automated culture systems

    Directory of Open Access Journals (Sweden)

    Diosdada Galvez Guerra

    2014-04-01

    Full Text Available One alternative to increase yields in the taro (Colocasia esculenta culture, could be the application of biotechnological methods, especially semi-automated systems. The new clone ‘INIVIT MC-2001’ with high productive potential, have low multiplication coefficients in conventional seed production systems. Therefore, the aim of this paper was to determine the effect of semi-automated systems on the in vitro multiplication of taro ‘INIVIT MC-2001’. The effect of three semi-automated systems culture was studied: temporary Immersion system, immersion system with constant aeration to the culture medium and constant immersion system with aeration to the inner atmosphere of the culture recipient. The type of semi-automated culture system influenced the multiplication coefficient, the best results were obtained with the temporary immersion system (8.46 and the immersion system with constant aeration of the culture medium (8.42 which are alternatives for increasing the multiplication coefficient in clone ‘INIVIT MC-2001’. Key words: in vitro culture, taro, temporary immersion

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

    Science.gov (United States)

    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

  18. Quantum cloning

    Energy Technology Data Exchange (ETDEWEB)

    Buzek, Vladimir [Slovak Academy of Sciences (Slovakia); Hillery, Mark [City University of New York (United States)

    2001-11-01

    It is impossible to make perfect copies or 'clones' of unknown quantum states, but approximate copies could still have many uses in quantum computing. A computer is a physical device that consists of components that are all subject to the laws of physics. Since computers deal exclusively in information, there is a close connection between information and physical systems. But what happens if the components inside the computer become so small that they must be described by quantum mechanics rather than classical physics? The seemingly unstoppable decrease in the size of transistors and other components will force the computer industry to confront this question in the near future. However, a small band of far-sighted physicists has been thinking about these problems for almost two decades. Starting with the work of Paul Benioff, Richard Feynman, David Deutsch and Charles Bennett in the mid-1980s, the field of 'quantum information' has grown to become one of the most exciting areas of modern physics. These early pioneers realized that the representation of information by quantum systems, such as single electrons or photons, was an opportunity rather than a problem. (U.K.)

  19. A novel high-throughput (HTP) cloning strategy for site-directed designed chimeragenesis and mutation using the Gateway cloning system.

    Science.gov (United States)

    Suzuki, Yasuhiro; Kagawa, Naoko; Fujino, Toru; Sumiya, Tsuyoshi; Andoh, Taichi; Ishikawa, Kumiko; Kimura, Rie; Kemmochi, Kiyokazu; Ohta, Tsutomu; Tanaka, Shigeo

    2005-07-11

    There is an increasing demand for easy, high-throughput (HTP) methods for protein engineering to support advances in the development of structural biology, bioinformatics and drug design. Here, we describe an N- and C-terminal cloning method utilizing Gateway cloning technology that we have adopted for chimeric and mutant genes production as well as domain shuffling. This method involves only three steps: PCR, in vitro recombination and transformation. All three processes consist of simple handling, mixing and incubation steps. We have characterized this novel HTP method on 96 targets with >90% success. Here, we also discuss an N- and C-terminal cloning method for domain shuffling and a combination of mutation and chimeragenesis with two types of plasmid vectors.

  20. A host-vector system for gene cloning in the cyanobacterium Anacystis nidulans R2

    NARCIS (Netherlands)

    Kuhlemeier, C. J.; Thomas, A. A.; van der Ende, A.; van Leen, R. W.; Borrias, W. E.; van den Hondel, C. A.; van Arkel, G. A.

    1983-01-01

    We describe the construction of a series of vectors suitable for gene cloning in the cyanobacterium Anacystis nidulans R2. From the indigenous plasmid pUH24, derivatives were constructed with streptomycin as the selective marker; one of these plasmids was used to construct pUC303, a shuttle vector

  1. Human Cloning

    National Research Council Canada - National Science Library

    Johnson, Judith A; Williams, Erin D

    2006-01-01

    .... Scientists in other labs, including Harvard University and the University of California at San Francisco, intend to produce cloned human embryos in order to derive stem cells for medical research...

  2. BIOETHICS AND HUMAN CLONING

    Directory of Open Access Journals (Sweden)

    Željko Kaluđerović

    2011-12-01

    Full Text Available In this paper the authors analyze the process of negotiating and beginning of the United Nations Declaration on Human Cloning as well as the paragraphs of the very Declaration. The negotiation was originally conceived as a clear bioethical debate that should have led to a general agreement to ban human cloning. However, more often it had been discussed about human rights, cultural, civil and religious differences between people and about priorities in case of eventual conflicts between different value systems. In the end, a non-binding Declaration on Human Cloning had been adopted, full of numerous compromises and ambiguous formulations, that relativized the original intention of proposer states. According to authors, it would have been better if bioethical discussion and eventual regulations on cloning mentioned in the following text had been left over to certain professional bodies, and only after the public had been fully informed about it should relevant supranational organizations have taken that into consideration.

  3. Comprehensive clone screening and evaluation of fed-batch strategies in a microbioreactor and lab scale stirred tank bioreactor system: application on Pichia pastoris producing Rhizopus oryzae lipase

    Science.gov (United States)

    2014-01-01

    . pastoris Mut+ phenotype. The use of fed-batch strategies using mixed substrate feeds resulted in increased biomass and lipolytic activity. The automated processing of fed-batch strategies by the RoboLector considerably facilitates the operation of fermentation processes, while reducing error-prone clone selection by increasing product titers. The scale-up from microbioreactor to lab scale stirred tank bioreactor showed an excellent correlation, validating the use of microbioreactor as a powerful tool for evaluating fed-batch operational strategies. PMID:24606982

  4. Vestibular stimulation-induced facilitation of cervical premotoneuronal systems in humans.

    Directory of Open Access Journals (Sweden)

    Shinya Suzuki

    Full Text Available It is unclear how descending inputs from the vestibular system affect the excitability of cervical interneurons in humans. To elucidate this, we investigated the effects of galvanic vestibular stimulation (GVS on the spatial facilitation of motor-evoked potentials (MEPs induced by combined pyramidal tract and peripheral nerve stimulation. To assess the spatial facilitation, electromyograms were recorded from the biceps brachii muscles (BB of healthy subjects. Transcranial magnetic stimulation (TMS over the contralateral primary motor cortex and electrical stimulation of the ipsilateral ulnar nerve at the wrist were delivered either separately or together, with interstimulus intervals of 10 ms (TMS behind. Anodal/cathodal GVS was randomly delivered with TMS and/or ulnar nerve stimulation. The combination of TMS and ulnar nerve stimulation facilitated BB MEPs significantly more than the algebraic summation of responses induced separately by TMS and ulnar nerve stimulation (i.e., spatial facilitation. MEP facilitation significantly increased when combined stimulation was delivered with GVS (p < 0.01. No significant differences were found between anodal and cathodal GVS. Furthermore, single motor unit recordings showed that the short-latency excitatory peak in peri-stimulus time histograms during combined stimulation increased significantly with GVS. The spatial facilitatory effects of combined stimulation with short interstimulus intervals (i.e., 10 ms indicate that facilitation occurred at the premotoneuronal level in the cervical cord. The present findings therefore suggest that GVS facilitates the cervical interneuron system that integrates inputs from the pyramidal tract and peripheral nerves and excites motoneurons innervating the arm muscles.

  5. Facilitation Skills: The Catalyst for Increased Effectiveness in Consultant Practice and Clinical Systems Leadership

    Science.gov (United States)

    Manley, Kim; Titchen, Angie

    2017-01-01

    Consultant practitioner is the pinnacle of the clinical career ladder for all health care disciplines in the United Kingdom. Consultant nurse, midwife and health visitor roles build on the clinical credibility and expertise characteristic of advanced level practice, but also possess expertise in: clinical systems leadership and the facilitation of…

  6. Using Group Drawings Activities to Facilitate the Understanding of Systemic Aspects of Projects

    DEFF Research Database (Denmark)

    Arantes do Amaral, João Alberto; Hess, Aurélio; Gonçalves, Paulo

    2017-01-01

    technique, we followed the five-phased qualitative analysis method, combined with a systems analysis of the data obtained from observation. Five recurrent themes emerged: 1) Making drawings in groups helps content retention and facilitates connections between the concepts explained by the professor; 2...

  7. A novel approach to sequence validating protein expression clones with automated decision making

    Directory of Open Access Journals (Sweden)

    Mohr Stephanie E

    2007-06-01

    Full Text Available Abstract Background Whereas the molecular assembly of protein expression clones is readily automated and routinely accomplished in high throughput, sequence verification of these clones is still largely performed manually, an arduous and time consuming process. The ultimate goal of validation is to determine if a given plasmid clone matches its reference sequence sufficiently to be "acceptable" for use in protein expression experiments. Given the accelerating increase in availability of tens of thousands of unverified clones, there is a strong demand for rapid, efficient and accurate software that automates clone validation. Results We have developed an Automated Clone Evaluation (ACE system – the first comprehensive, multi-platform, web-based plasmid sequence verification software package. ACE automates the clone verification process by defining each clone sequence as a list of multidimensional discrepancy objects, each describing a difference between the clone and its expected sequence including the resulting polypeptide consequences. To evaluate clones automatically, this list can be compared against user acceptance criteria that specify the allowable number of discrepancies of each type. This strategy allows users to re-evaluate the same set of clones against different acceptance criteria as needed for use in other experiments. ACE manages the entire sequence validation process including contig management, identifying and annotating discrepancies, determining if discrepancies correspond to polymorphisms and clone finishing. Designed to manage thousands of clones simultaneously, ACE maintains a relational database to store information about clones at various completion stages, project processing parameters and acceptance criteria. In a direct comparison, the automated analysis by ACE took less time and was more accurate than a manual analysis of a 93 gene clone set. Conclusion ACE was designed to facilitate high throughput clone sequence

  8. CATO: The Clone Alignment Tool.

    Directory of Open Access Journals (Sweden)

    Peter V Henstock

    Full Text Available 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. Generating West Nile Virus from an Infectious Clone.

    Science.gov (United States)

    Vandergaast, Rianna; Fredericksen, Brenda L

    2016-01-01

    WNV infectious clones are valuable tools for elucidating WNV biology. Nevertheless, relatively few infectious WNV clones have been generated because their construction is hampered by the instability of flaviviral genomes. More recently, advances in cloning techniques as well as the development of several two-plasmid WNV infectious clone systems have facilitated the generation of WNV infectious clones. Here we described a protocol for recovering WNV from a two-plasmid system. In this approach, large quantities of these constructs are digested with restriction enzymes to produce complementary restriction sites at the 3' end of the upstream fragment and the 5' end of the downstream fragment. These fragments are then annealed to produce linear template for in vitro transcription to synthesize infectious RNA. The resulting RNA is transfected into cells and after several days WNV is recovered in the culture supernatant. This method can be used to generate virus from infectious clones encoding high- and low-pathogenicity strains of WNV, as well as chimeric virues.

  10. Intelligent Decisional Assistant that Facilitate the Choice of a Proper Computer System Applied in Busines

    Directory of Open Access Journals (Sweden)

    Nicolae MARGINEAN

    2009-01-01

    Full Text Available The choice of a proper computer system is not an easy task for a decider. One reason could be the present market development of computer systems applied in business. The big number of the Romanian market players determines a big number of computerized products, with a multitude of various properties. Our proposal tries to optimize and facilitate this decisional process within an e-shop where are sold IT packets applied in business, building an online decisional assistant, a special component conceived to facilitate the decision making needed for the selection of the pertinent IT package that fits the requirements of one certain business, described by the decider. The user interacts with the system as an online buyer that visit an e-shop where are sold IT package applied in economy.

  11. Golden GATEway cloning--a combinatorial approach to generate fusion and recombination constructs.

    Directory of Open Access Journals (Sweden)

    Stephan Kirchmaier

    Full Text Available The design and generation of DNA constructs is among the necessary but generally tedious tasks for molecular biologists and, typically, the cloning strategy is restricted by available restriction sites. However, increasingly sophisticated experiments require increasingly complex DNA constructs, with an intricacy that exceeds what is achievable using standard cloning procedures. Many transgenes such as inducible gene cassettes or recombination elements consist of multiple components that often require precise in-frame fusions. Here, we present an efficient protocol that facilitates the generation of these complex constructs. The golden GATEway cloning approach presented here combines two established cloning methods, namely golden Gate cloning and Multisite Gateway(TM cloning. This allows efficient and seamless assembly as well as reuse of predefined DNA elements. The golden Gate cloning procedure follows clear and simple design rules and allows the assembly of multiple fragments with different sizes into one open reading frame. The final product can be directly integrated into the widely used Multisite Gateway(TM cloning system, granting more flexibility when using a transgene in the context of multiple species. This adaptable and streamlined cloning procedure overcomes restrictions of "classical construct generation" and allows focusing on construct design.

  12. Golden GATEway Cloning – A Combinatorial Approach to Generate Fusion and Recombination Constructs

    Science.gov (United States)

    Wittbrodt, Joachim

    2013-01-01

    The design and generation of DNA constructs is among the necessary but generally tedious tasks for molecular biologists and, typically, the cloning strategy is restricted by available restriction sites. However, increasingly sophisticated experiments require increasingly complex DNA constructs, with an intricacy that exceeds what is achievable using standard cloning procedures. Many transgenes such as inducible gene cassettes or recombination elements consist of multiple components that often require precise in-frame fusions. Here, we present an efficient protocol that facilitates the generation of these complex constructs. The golden GATEway cloning approach presented here combines two established cloning methods, namely golden Gate cloning and Multisite GatewayTM cloning. This allows efficient and seamless assembly as well as reuse of predefined DNA elements. The golden Gate cloning procedure follows clear and simple design rules and allows the assembly of multiple fragments with different sizes into one open reading frame. The final product can be directly integrated into the widely used Multisite GatewayTM cloning system, granting more flexibility when using a transgene in the context of multiple species. This adaptable and streamlined cloning procedure overcomes restrictions of “classical construct generation” and allows focusing on construct design. PMID:24116091

  13. Golden GATEway cloning--a combinatorial approach to generate fusion and recombination constructs.

    Science.gov (United States)

    Kirchmaier, Stephan; Lust, Katharina; Wittbrodt, Joachim

    2013-01-01

    The design and generation of DNA constructs is among the necessary but generally tedious tasks for molecular biologists and, typically, the cloning strategy is restricted by available restriction sites. However, increasingly sophisticated experiments require increasingly complex DNA constructs, with an intricacy that exceeds what is achievable using standard cloning procedures. Many transgenes such as inducible gene cassettes or recombination elements consist of multiple components that often require precise in-frame fusions. Here, we present an efficient protocol that facilitates the generation of these complex constructs. The golden GATEway cloning approach presented here combines two established cloning methods, namely golden Gate cloning and Multisite Gateway(TM) cloning. This allows efficient and seamless assembly as well as reuse of predefined DNA elements. The golden Gate cloning procedure follows clear and simple design rules and allows the assembly of multiple fragments with different sizes into one open reading frame. The final product can be directly integrated into the widely used Multisite Gateway(TM) cloning system, granting more flexibility when using a transgene in the context of multiple species. This adaptable and streamlined cloning procedure overcomes restrictions of "classical construct generation" and allows focusing on construct design.

  14. School-wide Systems to Promote Positive Behaviors and Facilitate Instruction

    OpenAIRE

    Lane, Kathleen Lynne; Menzies, Holly Mariah; Ennis, Robin Parks; Bezdek, Jamie

    2013-01-01

    In this article we focus on school-wide systems to promote positive behaviors and enhance instruction. Part of the appeal of multi-tiered systems of support is that they facilitate collaboration between teachers by clarifying school-wide goals and the means for achieving them. We begin with an overview of multi-tiered systems of support including: (a) a description of each level of prevention: primary (Tier 1, for all), secondary (Tier 2, for some), and tertiary (Tier 3, for a few); (b) inf...

  15. Modeling for Colloid and Chelator Facilitated Nuclide Transport in Radioactive Waste Disposal System

    International Nuclear Information System (INIS)

    Lee, Youn Myoung; Jeong, Jong Tae

    2010-08-01

    A modeling study and development of a total system performance assessment (TSPA) program template, by which assessment of safety and performance for a radioactive waste repository with normal and/or abnormal nuclide release cases can be made has been developed. Colloid and chelator facilitated transport that is believed to result for faster nuclide transport in various mediabothinthegeosphereandbiospherehas been evaluated deterministically and probabilistically to demonstrate the capability of the template developed through this study. To this end colloid and chelator facilitated nuclide transport has been modeled rather strainghtforwardly with assumed data through this study by utilizing some powerful function offered by GoldSim. An evaluation in view of apparent influence of colloid and chelator on the nuclide transport in the various media in and around a repository system with data assumed are illustrated

  16. Legal-institutional arrangements facilitating offshore wind energy conversion systems (WECS) utilization. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Mayo, L.H.

    1977-09-01

    Concern for the continuing sufficiency of energy supplies in the U.S. has tended to direct increasing attention to unconventional sources of supply, including wind energy. Some of the more striking proposals for the utilization of wind energy relate to offshore configurations. The legal-institutional arrangements for facilitating the utilization of offshore wind energy conversion systems (WECS) are examined by positioning three program alternatives and analyzing the institutional support required for the implementation of each.

  17. Spatial moment analysis of colloid facilitated radionuclide transport in a coupled fracture-matrix system

    OpenAIRE

    N. Natarajan, G. Suresh Kumar

    2011-01-01

    A numerical model is developed for studying the transport of colloid facilitated radionuclide transport in a coupled fracture-matrix system. The radionuclides and the colloids are assumed to decay, sorb on the fracture surface, as well as diffuse into the rock matrix. The sorption of the radionuclides onto the mobile and immobile colloids within the fracture is assumed to be linear. The governing equations describing the radionuclide and colloidal transport along the fracture axis and diffusi...

  18. Facilitating ambulatory electronic health record system implementation: evidence from a qualitative study.

    Science.gov (United States)

    McAlearney, Ann Scheck; Sieck, Cynthia; Hefner, Jennifer; Robbins, Julie; Huerta, Timothy R

    2013-01-01

    Ambulatory care practices have increasing interest in leveraging the capabilities of electronic health record (EHR) systems, but little information is available documenting how organizations have successfully implemented these systems. Objective. To characterize elements of successful electronic health record (EHR) system implementation and to synthesize the key informants' perspectives about successful implementation practices. Key informant interviews and focus groups were conducted with a purposive sample of individuals from US healthcare organizations identified for their success with ambulatory EHR implementation. Rigorous qualitative data analyses used both deductive and inductive methods. Participants identified personal and system-related barriers, at both the individual and organization levels, including poor computer skills, productivity losses, resistance to change, and EHR system failure. Implementation success was reportedly facilitated by careful planning and consistent communication throughout distinct stages of the implementation process. A significant element of successful implementation was an emphasis on optimization, both during "go-live" and, subsequently, when users had more experience with the system. Successful EHR implementation requires both detailed planning and clear mechanisms to deal with unforeseen or unintended consequences. Focusing on user buy-in early and including plans for optimization can facilitate greater success.

  19. Facilitating Ambulatory Electronic Health Record System Implementation: Evidence from a Qualitative Study

    Directory of Open Access Journals (Sweden)

    Ann Scheck McAlearney

    2013-01-01

    Full Text Available Background. Ambulatory care practices have increasing interest in leveraging the capabilities of electronic health record (EHR systems, but little information is available documenting how organizations have successfully implemented these systems. Objective. To characterize elements of successful electronic health record (EHR system implementation and to synthesize the key informants' perspectives about successful implementation practices. Methods. Key informant interviews and focus groups were conducted with a purposive sample of individuals from US healthcare organizations identified for their success with ambulatory EHR implementation. Rigorous qualitative data analyses used both deductive and inductive methods. Results. Participants identified personal and system-related barriers, at both the individual and organization levels, including poor computer skills, productivity losses, resistance to change, and EHR system failure. Implementation success was reportedly facilitated by careful planning and consistent communication throughout distinct stages of the implementation process. A significant element of successful implementation was an emphasis on optimization, both during “go-live” and, subsequently, when users had more experience with the system. Conclusion. Successful EHR implementation requires both detailed planning and clear mechanisms to deal with unforeseen or unintended consequences. Focusing on user buy-in early and including plans for optimization can facilitate greater success.

  20. SOFTICE: Facilitating both Adoption of Linux Undergraduate Operating Systems Laboratories and Students' Immersion in Kernel Code

    Directory of Open Access Journals (Sweden)

    Alessio Gaspar

    2007-06-01

    Full Text Available This paper discusses how Linux clustering and virtual machine technologies can improve undergraduate students' hands-on experience in operating systems laboratories. Like similar projects, SOFTICE relies on User Mode Linux (UML to provide students with privileged access to a Linux system without creating security breaches on the hosting network. We extend such approaches in two aspects. First, we propose to facilitate adoption of Linux-based laboratories by using a load-balancing cluster made of recycled classroom PCs to remotely serve access to virtual machines. Secondly, we propose a new approach for students to interact with the kernel code.

  1. Assessment of six tapping systems for four Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell.-Arg.,clones in the Colombian High Plains

    International Nuclear Information System (INIS)

    Quesada Mendez, Isaac; Aristizabal Gutierrez, Fabio Ancizar; Chaves Cordoba, Bernardo

    2009-01-01

    The rubber yield of four Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell.-Arg. clones was evaluated on the Colombian high plains in the Meta department: IAN 873 and RRIM 600 (third year of tapping being done) and PB 260 and GT 1 (first year of tapping). Six tapping systems were used, including a combination of different tapping frequencies (d/4 and d/5), Ethephon concentrations (0%, 2.5%, 3.3% and 5%) and a number of applications per year (4 to 8) were also used depending on the clone. The production figures for one commercial year were obtained from assays, using a completely random- block design (having four repetitions) independently defined for each clone: having on average 4446 g rubber/tree of produce per year for the RRIM 600 clone within a system 1/2S, d/4, 6d/7, 10m/12, ET 2.5%, Pa 7/y; 2696 g rubber/tree for the PB 260 clone with 1/2S, d/4, 6d/7, 10m/12, ET 2.5%, Pa 5/y; 3822 g rubber/tree for the IAN 873 clone, 1/2S, d/4, 6d/7, 10m/12, ET 3.3%, Pa 8/y, and 3472 g rubber/ tree for the GT 1 clone, 1/2S, d/4, 6d/7, 10m/12, ET 2.5%, Pa 5/y. The highest produce was obtained with a four-day tapping frequency.

  2. Facilitating mathematics learning for students with upper extremity disabilities using touch-input system.

    Science.gov (United States)

    Choi, Kup-Sze; Chan, Tak-Yin

    2015-03-01

    To investigate the feasibility of using tablet device as user interface for students with upper extremity disabilities to input mathematics efficiently into computer. A touch-input system using tablet device as user interface was proposed to assist these students to write mathematics. User-switchable and context-specific keyboard layouts were designed to streamline the input process. The system could be integrated with conventional computer systems only with minor software setup. A two-week pre-post test study involving five participants was conducted to evaluate the performance of the system and collect user feedback. The mathematics input efficiency of the participants was found to improve during the experiment sessions. In particular, their performance in entering trigonometric expressions by using the touch-input system was significantly better than that by using conventional mathematics editing software with keyboard and mouse. The participants rated the touch-input system positively and were confident that they could operate at ease with more practice. The proposed touch-input system provides a convenient way for the students with hand impairment to write mathematics and has the potential to facilitate their mathematics learning. Implications for Rehabilitation Students with upper extremity disabilities often face barriers to learning mathematics which is largely based on handwriting. Conventional computer user interfaces are inefficient for them to input mathematics into computer. A touch-input system with context-specific and user-switchable keyboard layouts was designed to improve the efficiency of mathematics input. Experimental results and user feedback suggested that the system has the potential to facilitate mathematics learning for the students.

  3. A decision support system in order to facilitate new financing actions in the public sector

    Directory of Open Access Journals (Sweden)

    Maria Franca Norese

    2014-12-01

    Full Text Available This analytical in-depth study on the financing modalities adopted in the Region of Piedmont was carried out with the contribution of those directly involved in the decision-making process enabling to acquire, codify and organize important elements of information which can be used to increase the evaluation culture in the Region. Owing to an increased interest in making public financing actions more efficient, the results of this analysis are of particular interest as they can be used as a multi-criteria evaluation model and a decision support system aimed at facilitating and implementing new financing and monitoring actions in a context of organizational learning.

  4. Using Group Drawings Activities to Facilitate the Understanding of the Systemic Aspects of Projects

    Directory of Open Access Journals (Sweden)

    João Alberto Arantes do Amaral

    2017-04-01

    Full Text Available In this article, we present our findings regarding promoting group drawing activities in order to facilitate the learning of systemic aspects of projects. We discuss the approach we used to engage the students and foster learning in our classes. We used group drawing activities in two project management undergraduate courses. The courses, which involved 41 students, took place during the second semester of 2016 in a public university in Brazil. We conducted qualitative research, using qualitative observation and focus group interviews. In order to gauge the effects of the use of this educational technique, we followed the five-phased qualitative analysis method, combined with a systems analysis of the data obtained from observation. Five recurrent themes emerged: 1 Making drawings in groups helps content retention and facilitates connections between the concepts explained by the professor; 2 Making drawings in groups promotes knowledge sharing among team members; 3 Making drawings in group fosters creativity and communication between students; 4 Drawing in groups reduces the students’ boredom, makes the lecture more dynamic and interesting; 5 Drawing in groups reinforces bonds between students. Our systems analysis suggests that group drawing improves student participation in classroom activities, strengthens bonds between students, and enhances learning.

  5. Implementation of Environmental Flows for Intermittent River Systems: Adaptive Management and Stakeholder Participation Facilitate Implementation

    Science.gov (United States)

    Conallin, John; Wilson, Emma; Campbell, Josh

    2018-03-01

    Anthropogenic pressure on freshwater ecosystems is increasing, and often leading to unacceptable social-ecological outcomes. This is even more prevalent in intermittent river systems where many are already heavily modified, or human encroachment is increasing. Although adaptive management approaches have the potential to aid in providing the framework to consider the complexities of intermittent river systems and improve utility within the management of these systems, success has been variable. This paper looks at the application of an adaptive management pilot project within an environmental flows program in an intermittent stream (Tuppal Creek) in the Murray Darling Basin, Australia. The program focused on stakeholder involvement, participatory decision-making, and simple monitoring as the basis of an adaptive management approach. The approach found that by building trust and ownership through concentrating on inclusiveness and transparency, partnerships between government agencies and landholders were developed. This facilitated a willingness to accept greater risks and unintended consequences allowing implementation to occur.

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

    Science.gov (United States)

    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.

  7. Facilitating innovations

    NARCIS (Netherlands)

    Buurma, J.S.; Visser, A.J.; Migchels, G.

    2011-01-01

    Many innovations involve changes which transcend the individual business or are only achievable when various businesses and/or interested parties take up the challenge together. In System Innovation Programmes, the necessary innovations are facilitated by means of workshops related to specific areas

  8. Home Automated Telemanagement (HAT System to Facilitate Self-Care of Patients with Chronic Diseases

    Directory of Open Access Journals (Sweden)

    Joseph Finkelstein

    2003-06-01

    Full Text Available Successful patient self-management requires a multidisciplinary approach that includes regular patient assessment, disease-specific education, control of medication adherence, implementation of health behavior change models and social support. Existing systems for computer-assisted disease management do not provide this multidisciplinary patient support and do not address treatment compliance issues. We developed the Home Automated Telemanagement (HAT system for patients with different chronic health conditions to facilitate their self-care. The HAT system consists of a home unit, HAT server, and clinician units. Patients at home use a palmtop or a laptop connected with a disease monitor on a regular basis. Each HAT session consists of self-testing, feedback, and educational components. The self-reported symptom data and objective results obtained from disease-specific sensors are automatically sent from patient homes to the HAT server in the hospital. Any web-enabled device can serve as a clinician unit to review patient results. The HAT system monitors self-testing results and patient compliance. The HAT system has been implemented and tested in patients receiving anticoagulation therapy, patients with asthma, COPD and other health conditions. Evaluation results indicated high level of acceptance of the HAT system by the patients and that the system has a positive impact on main clinical outcomes and patient satisfaction with medical care.

  9. Cloning and characterization of a P2X receptor expressed in the central nervous system of Lymnaea stagnalis.

    Directory of Open Access Journals (Sweden)

    Selvan Bavan

    Full Text Available P2X receptors are membrane ion channels gated by extracellular ATP. Mammals possess seven distinct P2X subtypes (P2X1-7 that have important functions in a wide array of physiological processes including roles in the central nervous system (CNS where they have been linked to modulation of neurotransmitter release. We report here the cloning and functional characterization of a P2X receptor from the mollusc Lymnaea stagnalis. This model organism has a relatively simple CNS consisting of large readily identifiable neurones, a feature which together with a well characterized neuronal circuitry for important physiological processes such as feeding and respiration makes it an attractive potential model to examine P2X function. Using CODEHOP PCR we identified a single P2X receptor (LymP2X in Lymnaea CNS which was subsequently cloned by RT-PCR. When heterologously expressed in Xenopus oocytes, LymP2X exhibited ATP evoked inward currents (EC(50 6.2 µM which decayed during the continued presence of agonist. UTP and ADP did not activate the receptor whereas αβmeATP was a weak agonist. BzATP was a partial agonist with an EC(50 of 2.4 µM and a maximal response 33% smaller than that of ATP. The general P2 receptor antagonists PPADS and suramin both inhibited LymP2X currents with IC(50 values of 8.1 and 27.4 µM respectively. LymP2X is inhibited by acidic pH whereas Zn(2+ and Cu(2+ ions exhibited a biphasic effect, potentiating currents up to 100 µM and inhibiting at higher concentrations. Quantitative RT-PCR and in situ hybridization detected expression of LymP2X mRNA in neurones of all CNS ganglia suggesting this ion channel may have widespread roles in Lymnaea CNS function.

  10. Secure the Clones

    Science.gov (United States)

    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.

  11. Statement on Human Cloning

    Science.gov (United States)

    ... for the Advancement of Science Statement on Human Cloning Tweet The American Association for the Advancement of ... for this statement on human cloning. Ban Reproductive Cloning AAAS endorses a legally enforceable ban on efforts ...

  12. Cloning and analysis of the genes encoding the type IIS restriction-modification system HphI from Haemophilus parahaemolyticus.

    Science.gov (United States)

    Lubys, A; Lubienè, J; Kulakauskas, S; Stankevicius, K; Timinskas, A; Janulaitis, A

    1996-07-15

    The genomic region encoding the type IIS restriction-modification (R-M) system HphI (enzymes recognizing the asymmetric sequence 5'-GGTGA-3'/5'-TCACC-3') from Haemophilus parahaemolyticus were cloned into Escherichia coli and sequenced. Sequence analysis of the R-M HphI system revealed three adjacent genes aligned in the same orientation: a cytosine 5 methyltransferase (gene hphIMC), an adenine N6 methyltransferase (hphIMA) and the HphI restriction endonuclease (gene hphIR). Either methyltransferase is capable of protecting plasmid DNA in vivo against the action of the cognate restriction endonuclease. hphIMA methylation renders plasmid DNA resistant to R.Hindill at overlapping sites, suggesting that the adenine methyltransferase modifies the 3'-terminal A residue on the GGTGA strand. Strong homology was found between the N-terminal part of the m6A methyltransferasease and an unidentified reading frame interrupted by an incomplete gaIE gene of Neisseria meningitidis. The HphI R-M genes are flanked by a copy of a 56 bp direct nucleotide repeat on each side. Similar sequences have also been identified in the non-coding regions of H.influenzae Rd DNA. Possible involvement of the repeat sequences in the mobility of the HphI R-M system is discussed.

  13. Improved cloning efficiency and developmental potential in bovine somatic cell nuclear transfer with the oosight imaging system.

    Science.gov (United States)

    Kim, Eun Young; Park, Min Jee; Park, Hyo Young; Noh, Eun Ji; Noh, Eun Hyung; Park, Kyoung Sik; Lee, Jun Beom; Jeong, Chang Jin; Riu, Key Zung; Park, Se Pill

    2012-08-01

    In somatic cell nuclear transfer (SCNT) procedures, exquisite enucleation of the recipient oocyte is critical to cloning efficiency. The purpose of this study was to compare the effects of two enucleation systems, Hoechst staining and UV irradiation (hereafter, irradiation group) and Oosight imaging (hereafter, Oosight group), on the in vitro production of bovine SCNT embryos. In the Oosight group, the apoptotic index (2.8 ± 0.5 vs. 7.3 ± 1.2) was lower, and the fusion rate (75.6% vs. 62.9%), cleavage rate (78.0% vs. 63.7%), blastocyst rate (40.2% vs. 29.2%), and total cell number (128.3±4.8 vs. 112.2 ± 7.6) were higher than those in the irradiation group (all p<0.05). The overall efficiency after SCNT was twice as high in the Oosight group as that in the irradiation group (p<0.05). The relative mRNA expression levels of Oct4, Nanog, Interferon-tau, and Dnmt3A were higher and those of Caspase-3 and Hsp70 were lower in the Oosight group compared with the irradiation group (p<0.05). This is the first report to show the positive effect of the Oosight imaging system on molecular gene expression in the SCNT embryo. The Oosight imaging system may become the preferred choice for enucleation because it is less detrimental to the developmental potential of bovine SCNT embryos.

  14. A System Concept for Facilitating User Preferences in En Route Airspace

    Science.gov (United States)

    Vivona, R. A.; Ballin, M. G.; Green, S. M.; Bach, R. E.; McNally, B. D.

    1996-01-01

    The Federal Aviation Administration is trying to make its air traffic management system more responsive to the needs of the aviation community by exploring the concept of 'free flight' for aircraft flying under instrument flight rules. A logical first step toward free flight could be made without significantly altering current air traffic control (ATC) procedures or requiring new airborne equipment by designing a ground-based system to be highly responsive to 'user preference' in en route airspace while providing for an orderly transition to the terminal area. To facilitate user preference in all en route environments, a system based on an extension of the Center/TRACON Automation System (CTAS) is proposed in this document. The new system would consist of two integrated components. An airspace tool (AT) focuses on unconstrained en route aircraft (e.g., not transitioning to the terminal airspace), taking advantage of the relatively unconstrained nature of their flights and using long-range trajectory prediction to provide cost-effective conflict resolution advisories to sector controllers. A sector tool (ST) generates efficient advisories for all aircraft, with a focus on supporting controllers in analyzing and resolving complex, highly constrained traffic situations. When combined, the integrated AT/ST system supports user preference in any air route traffic control center sector. The system should also be useful in evaluating advanced free-flight concepts by serving as a test bed for future research. This document provides an overview of the design concept, explains its anticipated benefits, and recommends a development strategy that leads to a deployable system.

  15. Estandarización de sistemas isoenzimáticos en clones colombianos del género musa Standarization of isozymic systems in clones of the colombian collection of the genus musa

    Directory of Open Access Journals (Sweden)

    Beltrán Margarita

    1996-12-01

    Full Text Available Con el propósito de estudiar la variabilidad genética presente en la Colección Colombiana de Musáceas, S8 estandarizaron varios sistemas enzimáticos en clones de diferentes ploidías. conservados bajo condiciones in vitro. Se probaron un total de 23 sistemas, seis de los cuales presentaron actividad electroforética en condiciones diferentes a las previamente reportadas por otros autores: Diaforasa (DIA E.C.1.6A.3., Enzima málica (ME E.C.l.1.1AO., Fosfoglucoisomerasa (PGI E.C.5.3.1.9., Fosfogluconato deshidrogenasa (PGDH E.C.1.1.1A4., Fosfoglucomutasa (PGM E.C.2.7.5.1. Y Rubisco (RUB E.CA.11.1.39. . Las enzimas DIA y RUB se reportan por primera vez para el genera Musa, ME, GDH, PGDH Y PGI se reportan por primera vez en geles de acrilamida. Diaforasa presentó alto pournornsmo en los clones probados, mientras que Rubisco mostró una sola zona de actividad.The main purpose of this study was the standardization of several isozymic systems using clones of different ploidy belonging to the Colombian Collection of Musa maintained under in vitro conditions. A total of 23 systems were assayed. Six isozymes presented new electrophoretic activity with regard to previous reports: Diaphorase (DIA E.C.1.6A.3., Malic enzyme(ME E.C.1.1.1AO Phosphoglucoisomerase (PGI E.C.5.3.1.9., Phosphogluconate dehydrogenase (PGDH E.C.1.1.1.44., Phosphoglucomutase (PGM E.C.2.7.5.1. and Rubisco (RUB E.CA.11.1.39.. The enzymes DIA and RUB are reported the first time for the genus Musa. ME, GDH, PGDH and PGI are described the first time in acrylamide support. DIA presented high level of polymorphism in the Colombian clones tested. RUB showed only one zone of activity.

  16. Systems and methods for facilitating hydrogen storage using naturally occurring nanostructure assemblies

    Science.gov (United States)

    Fliermans,; Carl, B [Augusta, GA

    2012-08-07

    Some or all of the needs above can be addressed by embodiments of the invention. According to embodiments of the invention, systems and methods for facilitating hydrogen storage using naturally occurring nanostructure assemblies can be implemented. In one embodiment, a method for storing hydrogen can be provided. The method can include providing diatoms comprising diatomaceous earth or diatoms from a predefined culture. In addition, the method can include heating the diatoms in a sealed environment in the presence of at least one of titanium, a transition metal, or a noble metal to provide a porous hydrogen storage medium. Furthermore, the method can include exposing the porous hydrogen storage medium to hydrogen. In addition, the method can include storing at least a portion of the hydrogen in the porous hydrogen storage medium.

  17. Molecular cloning of amphioxus uncoupling protein and assessment of its uncoupling activity using a yeast heterologous expression system

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Kun [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Sun, Guoxun [Department of Hematology, Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001 (China); Lv, Zhiyuan; Wang, Chen [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Jiang, Xueyuan, E-mail: xueyuanjiang@yahoo.com.cn [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Li, Donghai, E-mail: lidonghai@gmail.com [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Zhang, Chenyu, E-mail: cyzhang@nju.edu.cn [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China)

    2010-10-01

    Research highlights: {yields} Invertebrates, for example amphioxus, do express uncoupling proteins. {yields} Both the sequence and the uncoupling activity of amphioxus UCP resemble UCP2. {yields} UCP1 is the only UCP that can form dimer on yeast mitochondria. -- Abstract: The present study describes the molecular cloning of a novel cDNA fragment from amphioxus (Branchiostoma belcheri) encoding a 343-amino acid protein that is highly homologous to human uncoupling proteins (UCP), this protein is therefore named amphioxus UCP. This amphioxus UCP shares more homology with and is phylogenetically more related to mammalian UCP2 as compared with UCP1. To further assess the functional similarity of amphioxus UCP to mammalian UCP1 and -2, the amphioxus UCP, rat UCP1, and human UCP2 were separately expressed in Saccharomyces cerevisiae, and the recombinant yeast mitochondria were isolated and assayed for the state 4 respiration rate and proton leak, using pYES2 empty vector as the control. UCP1 increased the state 4 respiration rate by 2.8-fold, and the uncoupling activity was strongly inhibited by GDP, while UCP2 and amphioxus UCP only increased the state 4 respiration rate by 1.5-fold and 1.7-fold in a GDP-insensitive manner, moreover, the proton leak kinetics of amphioxus UCP was very similar to UCP2, but much different from UCP1. In conclusion, the amphioxus UCP has a mild, unregulated uncoupling activity in the yeast system, which resembles mammalian UCP2, but not UCP1.

  18. Molecular cloning of amphioxus uncoupling protein and assessment of its uncoupling activity using a yeast heterologous expression system

    International Nuclear Information System (INIS)

    Chen, Kun; Sun, Guoxun; Lv, Zhiyuan; Wang, Chen; Jiang, Xueyuan; Li, Donghai; Zhang, Chenyu

    2010-01-01

    Research highlights: → Invertebrates, for example amphioxus, do express uncoupling proteins. → Both the sequence and the uncoupling activity of amphioxus UCP resemble UCP2. → UCP1 is the only UCP that can form dimer on yeast mitochondria. -- Abstract: The present study describes the molecular cloning of a novel cDNA fragment from amphioxus (Branchiostoma belcheri) encoding a 343-amino acid protein that is highly homologous to human uncoupling proteins (UCP), this protein is therefore named amphioxus UCP. This amphioxus UCP shares more homology with and is phylogenetically more related to mammalian UCP2 as compared with UCP1. To further assess the functional similarity of amphioxus UCP to mammalian UCP1 and -2, the amphioxus UCP, rat UCP1, and human UCP2 were separately expressed in Saccharomyces cerevisiae, and the recombinant yeast mitochondria were isolated and assayed for the state 4 respiration rate and proton leak, using pYES2 empty vector as the control. UCP1 increased the state 4 respiration rate by 2.8-fold, and the uncoupling activity was strongly inhibited by GDP, while UCP2 and amphioxus UCP only increased the state 4 respiration rate by 1.5-fold and 1.7-fold in a GDP-insensitive manner, moreover, the proton leak kinetics of amphioxus UCP was very similar to UCP2, but much different from UCP1. In conclusion, the amphioxus UCP has a mild, unregulated uncoupling activity in the yeast system, which resembles mammalian UCP2, but not UCP1.

  19. A set of dual promoter vectors for high throughput cloning, screening, and protein expression in eukaryotic and prokaryotic systems from a single plasmid.

    Science.gov (United States)

    Sinah, Namita; Williams, Charlotte A; Piper, Robert C; Shields, S Brookhart

    2012-08-23

    The ability to produce the same recombinant protein in both prokaryotic and eukaryotic cells offers many experimental opportunities. However, the cloning of the same gene into multiple plasmids is required, which is time consuming, laborious and still may not produce soluble, stable protein in sufficient quantities. We have developed a set of expression vectors that allows for ligation-independent cloning and rapid functional screening for protein expression in both E. coli and S. cerevisiae. A set of expression vectors was made that can express the same open reading frame in E. coli (via the T7 phage promoter) and in S. cerevisiae (via the CUP1 or MET25 promoter). These plasmids also contain the essential elements for replication and selection in both cell types and have several advantages: they allow for cloning of genes by homologous recombination in yeast, protein expression can be determined before plasmid isolation and sequencing, and a GST-fusion tag is added to aid in soluble expression and purification. We have also included a TEV recognition site that allows for the specific cleavage of the fusion proteins to yield native proteins. The dual promoter vectors can be used for rapid cloning, expression, and purification of target proteins from both prokaryotic and eukaryotic systems with the ability to study post-translation modifications.

  20. The Clone Factory

    Science.gov (United States)

    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…

  1. Construction of a high-efficiency cloning system using the Golden Gate method and I-SceI endonuclease for targeted gene replacement in Bacillus anthracis.

    Science.gov (United States)

    Wang, Tiantian; Wang, Dongshu; Lyu, Yufei; Feng, Erling; Zhu, Li; Liu, Chunjie; Wang, Yanchun; Liu, Xiankai; Wang, Hengliang

    2018-02-10

    To investigate gene function in Bacillus anthracis, a high-efficiency cloning system is required with an increased rate of allelic exchange. Golden Gate cloning is a molecular cloning strategy allowing researchers to simultaneously and directionally assemble multiple DNA fragments to construct target plasmids using type IIs restriction enzymes and T4 DNA ligase in the same reaction system. Here, a B. anthracis S-layer protein EA1 allelic exchange vector was successfully constructed using the Golden Gate method. No new restriction sites were introduced into this knockout vector, and seamless assembly of the DNA fragments was achieved. To elevate the efficiency of homologous recombination between the allelic exchange vector and chromosomal DNA, we introduced an I-SceI site into the allelic exchange vector. The eag gene was successfully knocked out in B. anthracis using this vector. Simultaneously, the allelic exchange vector construction method was developed into a system for generating B. anthracis allelic exchange vectors. To verify the effectiveness of this system, some other allelic exchange vectors were constructed and gene replacements were performed in B. anthracis. It is speculated that this gene knockout vector construction system and high-efficiency targeted gene replacement using I-SceI endonuclease can be applied to other Bacillus spp. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Early growth of promising rubber tree clones of Hevea brasiliensis (Willd. ex A. Juss. Müll. Arg. in an agroforestry system in Caquetá, Colombia

    Directory of Open Access Journals (Sweden)

    Armando Sterling Cuéllar

    2015-07-01

    Full Text Available This study aimed to evaluate the early growth of three rubber tree clones (Hevea brasiliensis (FX 4098, FDR 5788, and control IAN 873 in four planting systems with cupuasu (Theobroma grandiflorum and banana trees (Musa AAB at two localities in Caquetá (Colombian Amazonia. A completely randomized block design with three replications and split-plot arrangement, analyzed with a general linear mixed model, was used at each locality. Total height (AT, trunk circumference (CT, canopy area (AC and leaf area index (IAF were evaluated one and two years after establishment of rubber trees. Trunk circumference was the variable most correlated with different growth indices (r ≥ 0.94. Four growth indices were strongly influenced by temporal and clonal variation and by planting system (p < 0.01. The only significant differences between the two localities were for AC (p < 0.05. System x clone interaction was significant for IAF (p < 0.01. After two years, the highest growth indices were observed in rubber tree clone FX 4098 (AT = 4.43 m; CT = 13.00 cm, AC = 4.62 m2; LAI = 1.37, and in two of the three agroforestry systems (AT ≥ 4.08 m, CT ≥ 11.43 cm, AC ≥ 4.01 m2; IAF ≥ 1.54 compared with the control system (monoculture.

  3. Systems to Harness Digital Footprint to Elucidate and Facilitate Ageing in Place.

    Science.gov (United States)

    Knight, Alissa; Fouyaxis, John; Jarrad, Geoff; Beski, Kinga; Cho, Gerald; Bidargaddi, Niranjan

    2018-01-01

    The rapid ageing of the population is a worldwide inexorable demographic transformation. At a time of immense social, political and economic change, the growing elderly population is at the forefront of global burden, placing an increasing strain on the federal, state, and local budgets. Many public policy responses to the impending ageing epidemic have begun, particularly with regards to dementia prevention and quality of life for older adults. However, to date, the fruition of such efforts remains to be discovered. Indeed, there is a need to find more novel and multifaceted ways of understanding the fragmentary changes and underlying mechanisms in the biopsychosocial contexts of ageing. Discovering better ways to measure these intricate domains will create better insight into how to improve clinical and public health information systems for the development of more personalisation support and services across the continuum of aged care. Technology now permeates all aspects of our everyday living. Digital footprints are data arising as a by-product of interactions we do as part of everyday living. The digital traces we live behind, be it on internet, social media, on mobile phone apps, as well as in health records (EHRs) could be used to infer how we behave and interact with environment, and how we feel in different situations. Commercial sector has very successfully used these footprints in the advertisement and marketing space. This type of information may provide clinicians with an unobtrusive way of monitoring older adults in their daily living, and provide an alternative means to traditional self-report and expert-rated assessment. In this paper we present two innovative digital footprint applications, Actionable Intime Insights and the SAIL Mobile app, which aim to facilitate "Ageing in Place" through adaptive, dynamic, early intervention strategies. These systems are devised to unveil contextual indicators of how a person is functioning mentally, socially

  4. How to facilitate the practical application of Quality Systems in Nuclear Medicine

    International Nuclear Information System (INIS)

    Touzet, R.E.

    2002-01-01

    Aim: Which are the reasons for non-implementation of a Quality System in most of the nuclear medicine services in Latin America, in spite of the recommendations made by international agencies and of the absolute convenience involved in guaranteeing the quality of the health-care services rendered to the population? One of the projects of the Argentine Regulatory Authority was providing an answer to this question and finding the solution to the existing situation in a progressive manner. Materials and Methods: Within a set of about 400 NM service providers and with the support of the Argentine Association of Nuclear Medicine and Biology, an investigation was made in order to learn which were the opinions of their officials and staff (interviews, surveys and discussion meetings were performed). On the basis of the results, a strategy was developed involving the design of a 'basic quality system' to be used as an intermediate stage between the 'natural quality systems' and the 'certifiable quality systems', while the general requirements of the ISO 9000-2000 Standard were applied as a structure. Once an agreement was reached on the working program, several documents were widely distributed (Explanatory Guide, Format of the Quality Manual, etc.). Also, courses and practical workshops were carried out and advice was provided by e-mail. Currently, several additional documents are being developed in order to facilitate implementation, training and quality systems evaluation. Results: Some of the reasons for the scarce certification appeared to be as follows: The cost of certification is high and specialized advisors are required. The long time required causes disappointment. There are no economic stimuli because competition is based on costs and not on quality. Medical professionals are not trained in quality management during their graduate studies. The formal certification system does not allow for intermediate stages and is highly dependent on the contracted advisor

  5. Quantum cloning with multicopy in d-dimensions

    Science.gov (United States)

    Zhang, Wenhai; Yu, Longbao; Yang, Ming; Cao, Zhuoliang

    2011-12-01

    The explicit transformations of the 1 → 3 optimal universal quantum cloning and the optimal phase-covariant quantum cloning in d-dimensions are presented, and the dimensionalities of their ancillary systems are both d-dimensions. As d→∞, their clone fidelities move toward 1/3, showing a classical limit for the fidelity of quantum cloning. Based on the reduction of the unitary transformation of quantum cloning, the transformation of the 1→ M= d+1 optimal economical phase-covariant quantum cloning in d-dimensions is derived, and the clone fidelity is covered by the theoretical value.

  6. Development of the gateway recycling cloning system for multiple linking of expression cassettes in a defined order, and direction on gateway compatible binary vectors.

    Science.gov (United States)

    Kimura, Tetsuya; Nakao, Akihide; Murata, Sachiko; Kobayashi, Yasuyuki; Tanaka, Yuji; Shibahara, Kenta; Kawazu, Tetsu; Nakagawa, Tsuyoshi

    2013-01-01

    We developed the Gateway recycling cloning system, which allows multiple linking of expression cassettes by multiple rounds of the Gateway LR reaction. Employing this system, the recycling donor vector pRED419 was subjected to the first LR reaction with an attR1-attR2 type destination vector. Then conversion vector pCON was subjected to an LR reaction to restore the attR1-attR2 site on the destination vector for the next cloning cycle. By repetition of these two simple steps, we linked four expression cassettes of a reporter gene in Gateway binary vector pGWB1, introduced the constructs into tobacco BY-2 cells, and observed the expression of transgenes.

  7. Iron uptake system mediates nitrate-facilitated cadmium accumulation in tomato (Solanum lycopersicum) plants.

    Science.gov (United States)

    Luo, Bing Fang; Du, Shao Ting; Lu, Kai Xing; Liu, Wen Jing; Lin, Xian Yong; Jin, Chong Wei

    2012-05-01

    Nitrogen (N) management is a promising agronomic strategy to minimize cadmium (Cd) contamination in crops. However, it is unclear how N affects Cd uptake by plants. Wild-type and iron uptake-inefficient tomato (Solanum lycopersicum) mutant (T3238fer) plants were grown in pH-buffered hydroponic culture to investigate the direct effect of N-form on Cd uptake. Wild-type plants fed NO₃⁻ accumulated more Cd than plants fed NH₄⁺. Iron uptake and LeIRT1 expression in roots were also greater in plants fed NO₃⁻. However, in mutant T3238fer which loses FER function, LeIRT1 expression in roots was almost completely terminated, and the difference between NO₃⁻ and NH₄⁺ treatments vanished. As a result, the N-form had no effect on Cd uptake in this mutant. Furthermore, suppression of LeIRT1 expression by NO synthesis inhibition with either tungstate or L-NAME, also substantially inhibited Cd uptake in roots, and the difference between N-form treatments was diminished. Considering all of these findings, it was concluded that the up-regulation of the Fe uptake system was responsible for NO₃⁻-facilitated Cd accumulation in plants.

  8. Facilitating the improved management of waste in South Africa through a national waste information system

    International Nuclear Information System (INIS)

    Godfrey, Linda

    2008-01-01

    Developing a waste information system (WIS) for a country is more than just about collecting routine data on waste; it is about facilitating the improved management of waste by providing timely, reliable information to the relevant role-players. It is a means of supporting the waste governance challenges facing South Africa - challenges ranging from strategic waste management issues at national government to basic operational challenges at local government. The paper addresses two hypotheses. The first is that the identified needs of government can provide a platform from which to design a national WIS framework for a developing country such as South Africa, and the second is that the needs for waste information reflect greater, currently unfulfilled challenges in the sustainable management of waste. Through a participatory needs analysis process, it is shown that waste information is needed by the three spheres of government, to support amongst others, informed planning and decision-making, compliance monitoring and enforcement, community participation through public access to information, human, infrastructure and financial resource management and policy development. These needs for waste information correspond closely with key waste management challenges currently facing the country. A shift in governments approach to waste, in line with national and international policy, is evident from identified current and future waste information needs. However, the need for information on landfilling remains entrenched within government, possibly due to the poor compliance of landfill sites in South Africa and the problems around the illegal disposal of both general and hazardous waste

  9. Systemic dizocilpine (MK-801 facilitates performance in opposition to response bias

    Directory of Open Access Journals (Sweden)

    Lauwereyns Johan

    2007-09-01

    Full Text Available Abstract Previous research has established that dopamine signals are crucial in orienting behavior to reward. Less is known, however, about the psychopharmacology of task performance under small-reward conditions as compared to large-reward conditions. The current study examined the effects of the noncompetitive N-methyl-D-aspartate (NMDA-receptor antagonist dizocilpine (MK-801 on reaction time (RT in a nose-poke task with rats completing an asymmetric reward schedule. In all trials, the rats were required to poke their nose in either the left or the right peripheral hole immediately adjacent to the centre hole when the corresponding light was illuminated. Depending on the stimulus-reward mapping, however, one position was associated with a large reward, while the alternative position was associated with a small reward. Correct performance was required in every trial; if the rat did not make a correct response within 20 s, the trial was aborted, and the same stimulus was presented again on the next trial. In this way, the rat was forced to perform the same visuo-spatial discrimination task under different reward conditions. Reaction times (ms were faster for large-reward trials than for small-reward trials, replicating previous findings. At a dosage of MK-801 (0.04 mg/kg, there was no significant influence of on RT in large-reward trials. In contrast, the same dosage of MK-801 in small-reward trials produced a decrease in RT as compared to the control condition, implying an improvement of performance. Below 0.04 mg/kg of MK-801, a steady decrease of RT in small-trials was seen as a function of dosage. Above 0.04 mg/kg of MK-801, the majority of rats failed to perform the task at all, whereas the rats that did manage to perform the criterion of 80 correct trials in a session showed no difference in RT between large- and small-reward trials. These data indicate that the systemic administration of a relatively small dosage of MK-801 facilitates

  10. Remote sensing of rivers: an emerging tool to facilitate management and restoration of fluvial systems

    Science.gov (United States)

    Legleiter, C. J.; Overstreet, B. T.

    2013-12-01

    All phases of river restoration, from design to implementation to assessment, require spatially distributed, high-resolution data on channels and floodplains. Conventional field methods are cost prohibitive for large areas, but remote sensing presents an increasingly viable alternative for characterizing fluvial systems. For example, bathymetric maps useful for habitat assessment can be derived from readily available, free or low cost image data, provided depth measurements are available for calibration. In combination with LiDAR, spectrally-based bathymetry can be used to determine bed elevations for estimating scour and fill and/or to obtain topographic input data for morphodynamic modeling. New, water-penetrating green LiDAR systems that measure sub-aerial and submerged elevations could provide a single-sensor solution for mapping riparian environments. Our current research on the Snake River focuses on comparing optical- and LiDAR-based methods for retrieving depths and bed elevations. Multi-sensor surveys from 2012 and 2013 will allow us to evaluate each instrument's capabilities for measuring volumes of erosion and deposition in a dynamic gravel-bed river. Ongoing studies also suggest that additional river attributes, such as substrate composition and flow velocity, could be inferred from hyperspectral image data. In general, remote sensing has considerable potential to facilitate various aspects of river restoration, from site evaluation to post-project assessment. Moreover, by providing more extensive coverage, this approach favors an integrated, watershed perspective for planning, execution, and monitoring of sustainable restoration programs. To stimulate progress toward these objectives, our research group is now working to advance the remote sensing of rivers through tool development and sensor deployment. Bathymetric map of the Snake River, WY, derived from hyperspectral image data via optimal band ratio analysis. Flow direction is from right to left.

  11. Determining Barriers and Facilitators Associated With Willingness to Use a Personal Health Information Management System to Support Worksite Wellness Programs.

    Science.gov (United States)

    Neyens, David M; Childers, Ashley Kay

    2017-07-01

    To determine the barriers and facilitators associated with willingness to use personal health information management (PHIM) systems to support an existing worksite wellness program (WWP). The study design involved a Web-based survey. The study setting was a regional hospital. Hospital employees comprised the study subjects. Willingness, barriers, and facilitators associated with PHIM were measured. Bivariate logit models were used to model two binary dependent variables. One model predicted the likelihood of believing PHIM systems would positively affect overall health and willingness to use. Another predicted the likelihood of worrying about online security and not believing PHIM systems would benefit health goals. Based on 333 responses, believing PHIM systems would positively affect health was highly associated with willingness to use PHIM systems (p systems. Participants in exercise-based components of WWPs were 3.03 times more likely to be willing to use PHIM systems. Those who worried about online security were 5.03 times more likely to believe PHIM systems would not help obtain health goals. Comfort with personal health information online and exercise-based WWP experience was associated with willingness to use PHIM systems. However, nutrition-based WWPs did not have similar effects. Implementation barriers relate to technology anxiety and trust in security, as well as experience with specific WWP activities. Identifying differences between WWP components and addressing technology concerns before implementation of PHIM systems into WWPs may facilitate improved adoption and usage.

  12. Oxytocin-Oxytocin Receptor Systems Facilitate Social Defeat Posture in Male Mice.

    Science.gov (United States)

    Nasanbuyan, Naranbat; Yoshida, Masahide; Takayanagi, Yuki; Inutsuka, Ayumu; Nishimori, Katsuhiko; Yamanaka, Akihiro; Onaka, Tatsushi

    2018-02-01

    Social stress has deteriorating effects on various psychiatric diseases. In animal models, exposure to socially dominant conspecifics (i.e., social defeat stress) evokes a species-specific defeat posture via unknown mechanisms. Oxytocin neurons have been shown to be activated by stressful stimuli and to have prosocial and anxiolytic actions. The roles of oxytocin during social defeat stress remain unclear. Expression of c-Fos, a marker of neuronal activation, in oxytocin neurons and in oxytocin receptor‒expressing neurons was investigated in mice. The projection of oxytocin neurons was examined with an anterograde viral tracer, which induces selective expression of membrane-targeted palmitoylated green fluorescent protein in oxytocin neurons. Defensive behaviors during double exposure to social defeat stress in oxytocin receptor‒deficient mice were analyzed. After social defeat stress, expression of c-Fos protein was increased in oxytocin neurons of the bed nucleus of the stria terminalis, supraoptic nucleus, and paraventricular hypothalamic nucleus. Expression of c-Fos protein was also increased in oxytocin receptor‒expressing neurons of brain regions, including the ventrolateral part of the ventromedial hypothalamus and ventrolateral periaqueductal gray. Projecting fibers from paraventricular hypothalamic oxytocin neurons were found in the ventrolateral part of the ventromedial hypothalamus and in the ventrolateral periaqueductal gray. Oxytocin receptor‒deficient mice showed reduced defeat posture during the second social defeat stress. These findings suggest that social defeat stress activates oxytocin-oxytocin receptor systems, and the findings are consistent with the view that activation of the oxytocin receptor in brain regions, including the ventrolateral part of the ventromedial hypothalamus and the ventrolateral periaqueductal gray, facilitates social defeat posture.

  13. A yeast recombination-based cloning system to produce chimeric HIV-1 viruses and express HIV-1 genes.

    Science.gov (United States)

    Moore, Dawn M; Arts, Eric J; Gao, Yong; Marozsan, Andre J

    2005-01-01

    Differential phenotypes or properties of HIV-1 gene products in primary virus isolates are difficult to assess due to interference by the high degree of sequence variation across the entire genome. Thus, chimeric viruses provide a powerful tool to study the function of single gene products or genetic elements in the context of a neutral viral genomic backbone. In this chapter, we describe how to produce HIV-1 chimeric viruses utilizing a yeast-based homologous recombination cloning technique to insert env sequences first into a yeast cloning vector and then into the common pNL4-3 virus backbone. This technique is not limited to the env gene, but can be used to build chimeric viruses with any HIV-1 gene or genetic element. This cloning technique involves the use of a shuttle vector that can replicate in yeast and bacterial cells. Along with acting as a shuttle vector for subsequent subcloning into pNL4-3, this construct pRec/env can also be used to express to the env gene product, gp120/gp41, on the surface of mammalian cells. The chimeric viruses produced by this cloning method are capable of undergoing multiple rounds of replication and are therefore very useful to study drug sensitivity, coreceptor usage, and viral fitness as influenced by a single gene or gene fragment of a primary HIV-1 isolate from any group M subtype.

  14. A dual tag system for facilitated detection of surface expressed proteins in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Jarmander Johan

    2012-09-01

    Full Text Available Abstract Background The discovery of the autotransporter family has provided a mechanism for surface expression of proteins in laboratory strains of Escherichia coli. We have previously reported the use of the AIDA-I autotransport system to express the Salmonella enterica serovar Enteritidis proteins SefA and H:gm. The SefA protein was successfully exposed to the medium, but the orientation of H:gm in the outer membrane could not be determined due to proteolytic cleavage of the N-terminal detection-tag. The goal of the present work was therefore to construct a vector containing elements that facilitates analysis of surface expression, especially for proteins that are sensitive to proteolysis or otherwise difficult to express. Results The surface expression system pAIDA1 was created with two detection tags flanking the passenger protein. Successful expression of SefA and H:gm on the surface of E. coli was confirmed with fluorescently labeled antibodies specific for the N-terminal His6-tag and the C-terminal Myc-tag. While both tags were detected during SefA expression, only the Myc-tag could be detected for H:gm. The negative signal indicates a proteolytic cleavage of this protein that removes the His6-tag facing the medium. Conclusions Expression levels from pAIDA1 were comparable to or higher than those achieved with the formerly used vector. The presence of the Myc- but not of the His6-tag on the cell surface during H:gm expression allowed us to confirm the hypothesis that this fusion protein was present on the surface and oriented towards the cell exterior. Western blot analysis revealed degradation products of the same molecular weight for SefA and H:gm. The size of these fragments suggests that both fusion proteins have been cleaved at a specific site close to the C-terminal end of the passenger. This proteolysis was concluded to take place either in the outer membrane or in the periplasm. Since H:gm was cleaved to a much greater extent

  15. Local cloning of entangled states

    International Nuclear Information System (INIS)

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

    2010-01-01

    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.

  16. Identifying Facilitators and Barriers for Patient Safety in a Medicine Label Design System Using Patient Simulation and Interviews

    DEFF Research Database (Denmark)

    Dieckmann, Peter; Clemmensen, Marianne Hald; Sørensen, Trine Kart

    2016-01-01

    Objectives Medicine label design plays an important role in improving patient safety. This study aimed at identifying facilitators and barriers in a medicine label system to prevent medication errors in clinical use by health care professionals. Methods The study design is qualitative and explora...

  17. SLiCE: a novel bacterial cell extract-based DNA cloning method

    Science.gov (United States)

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2012-01-01

    We describe a novel cloning method termed SLiCE (Seamless Ligation Cloning Extract) 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 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from Bacteria Artificial Chromosomes (BACs) or other sources. SLiCE is highly cost effective as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. In addition, the cloning efficiencies and capabilities of these strains can be greatly improved by simple genetic modifications. As an example, we modified the DH10B Escherichia coli strain to express an optimized λ prophage Red recombination system. This strain, termed PPY, facilitates SLiCE with very high efficiencies and demonstrates the versatility of the method. PMID:22241772

  18. SLiCE: a novel bacterial cell extract-based DNA cloning method.

    Science.gov (United States)

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2012-04-01

    We describe a novel cloning method termed SLiCE (Seamless Ligation Cloning Extract) 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 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from Bacteria Artificial Chromosomes (BACs) or other sources. SLiCE is highly cost effective as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. In addition, the cloning efficiencies and capabilities of these strains can be greatly improved by simple genetic modifications. As an example, we modified the DH10B Escherichia coli strain to express an optimized λ prophage Red recombination system. This strain, termed PPY, facilitates SLiCE with very high efficiencies and demonstrates the versatility of the method.

  19. Report on Animal Cloning

    OpenAIRE

    Chrenek, Peter

    2008-01-01

    The importance of creation of clones is exhibited in attempts to conserve and reproduce genetically valuable animals (meaning of reproductive cloning) and to produce embryonic stem cells (meaning of therapeutic cloning). Further possibility of application of genetically identical individuals is their use in experiments for the study of environmental influences (nutrition, ethology). Other perspective usage of clones can be creation of genetically modified individuals (transgenesis) and in fie...

  20. Quantum cloning and signaling

    International Nuclear Information System (INIS)

    Simon, C.; Weihs, G.; Zeilinger, A.

    1999-01-01

    We discuss the close connections between cloning of quantum states and superluminal signaling. We present an optimal universal cloning machine based on stimulated emission recently proposed by the authors. As an instructive example, we show how a scheme for superluminal communication based on this cloning machine fails. (Authors)

  1. Cloning and expression of ligand-gated ion-channel receptor L2 in central nervous system

    International Nuclear Information System (INIS)

    Houtani, Takeshi; Munemoto, Yumi; Kase, Masahiko; Sakuma, Satoru; Tsutsumi, Toshiyuki; Sugimoto, Tetsuo

    2005-01-01

    An orphan receptor of ligand-gated ion-channel type (L2, also termed ZAC according to the presence of zinc ion for channel activation) was identified by computer-assisted search programs on human genome database. The L2 protein shares partial homology with serotonin receptors 5HT3A and 5HT3B. We have cloned L2 cDNA derived from human caudate nucleus and characterized the exon-intron structure as follows: (1) The L2 protein has four transmembrane regions (M1-M4) and a long cytoplasmic loop between M3 and M4. (2) The sequence is conserved in species including chimpanzee, dog, cow, and opossum. (3) Nine exons form its protein-coding region and especially exon 5 corresponds to a disulfide bond region on the amino-terminal side. Our analysis using multiple tissue cDNA panels revealed that at least two splicing variants of L2 mRNA are present. The cDNA PCR amplification study revealed that L2 mRNA is expressed in tissues including brain, pancreas, liver, lung, heart, kidney, and skeletal muscle while 5HT3A mRNA could be detected in brain, heart, placenta, lung, kidney, pancreas, and skeletal muscle, and 5HT3B mRNA in brain, kidney, and skeletal muscle, suggesting different significance in tissue expression of these receptors. Regional expression of L2 mRNA and protein was examined in brain. The RT-PCR studies confirmed L2 mRNA expression in hippocampus, striatum, amygdala, and thalamus in adult brain. The L2 protein was immunolocalized by using antipeptide antibodies. Immunostained tissue sections revealed that L2-like immunoreactivity was dominantly expressed in the hippocampal CA3 pyramidal cells and in the polymorphic layer of the dentate gyrus. We analyzed the expression of L2 protein in HEK293 cells using GFP fusion protein reporter system. Western blots revealed that L2 protein confers sugar chains on the extracellular side. In transfected HEK293 cells, cellular membranes and intracellular puncta were densely labeled with GFP, suggesting selective dispatch to the

  2. Pupillometric evidence for the locus coeruleus-noradrenaline system facilitating attentional processing of action-triggered visual stimuli

    Directory of Open Access Journals (Sweden)

    Ken eKihara

    2015-06-01

    Full Text Available It has been argued that attentional processing of visual stimuli is facilitated by a voluntary action that triggers the stimulus onset. However, the relationship between action-induced facilitation of attention and the neural substrates has not been well established. The present study investigated whether the locus coeruleus-noradrenaline (LC-NA system is involved in this facilitation effect. A rapid serial visual presentation paradigm was used to assess the dynamics of transient attention in humans. Participants were instructed to change a digit stream to a letter stream by pressing a button and specifying successive targets of four letters. Pupil dilation was measured as an index of LC-NA function. Accuracy of target identification was better when the temporal delay between participants' key press and target onset was 800 ms than when targets appeared just after the key press or when targets appeared without key press. Accuracy of target identification was positively correlated with both the peak amplitude of pupil dilation and the pupil size at the time of the key press. These results indicate that target identification in the visual task is closely linked to pupil dilation. We conclude that the LC-NA system plays an important role in the facilitation of transient attention driven by voluntary action.

  3. Systemic blockade of D2-like dopamine receptors facilitates extinction of conditioned fear in mice

    OpenAIRE

    Ponnusamy, Ravikumar; Nissim, Helen A.; Barad, Mark

    2005-01-01

    Extinction of conditioned fear in animals is the explicit model of behavior therapy for human anxiety disorders, including panic disorder, obsessive-compulsive disorder, and post-traumatic stress disorder. Based on previous data indicating that fear extinction in rats is blocked by quinpirole, an agonist of dopamine D2 receptors, we hypothesized that blockade of D2 receptors might facilitate extinction in mice, while agonists should block extinction, as they do in rats. One day after fear con...

  4. Facilitating Learning and Physical Change in Complex Systems through Employee Involvement

    DEFF Research Database (Denmark)

    Bjerrum, Eva; Dahl, Susanne

    In a Danish workplace an experiment with mobile seating was carried out. Instead of implementing a certain concept designed by the management team the process was facilitated as a user involvement process based on Stacey´s theory of complex responsive processes. Here providing alternative pictures...... of the organisation challenged the discursive practice of the organisation and engaged employees in a process where they challenged each other’s accepted understandings of the organisation and of their work....

  5. Rapid customised operon assembly by yeast recombinational cloning.

    Science.gov (United States)

    Liu, Michael A; Kenyon, Johanna J; Lee, Jason; Reeves, Peter R

    2017-06-01

    We have developed a system called the Operon Assembly Protocol (OAP), which takes advantage of the homologous recombination DNA repair pathway in Saccharomyces cerevisiae to assemble full-length operons from a series of overlapping PCR products into a specially engineered yeast-Escherichia coli shuttle vector. This flexible, streamlined system can be used to assemble several operon clones simultaneously, and each clone can be expressed in the same E. coli tester strain to facilitate direct functional comparisons. We demonstrated the utility of the OAP by assembling and expressing a series of E. coli O1A O-antigen gene cluster clones containing various gene deletions or replacements. We then used these constructs to assess the substrate preferences of several Wzx flippases, which are responsible for translocation of oligosaccharide repeat units (O units) across the inner membrane during O-antigen biosynthesis. We were able to identify several O unit structural features that appear to be important determinants of Wzx substrate preference. The OAP system should be broadly applicable for the genetic manipulation of any bacterial operon and can be modified for use in other host species. It could also have potential uses in fields such as glycoengineering.

  6. [A study on facilitators and inhibitors to the introduction of outsourcing in the hospital information systems in Korea].

    Science.gov (United States)

    Choy, Soon; Shin, Hyeong-Sik; Choi, Inyoung; Kim, Sukil

    2007-01-01

    This study was conducted to investigate the current status of outsourcing in Korean hospital information systems and the factors influencing its introduction. The authors surveyed 136 hospitals located in Seoul and its surrounding vicinities from June 7 to June 23, 2006. The facilitators and inhibitors to outsourcing in hospital information systems were derived from literature and expert reviews. Multiple logistic regression analysis was applied to identify the major influencing factors on outsourcing in hospital information systems. Eighty-six (63.2%) of the 136 hospitals surveyed, which were mainly tertiary hospitals, responded to using outsourcing for their hospital information systems. "Hardware and software maintenance and support," "application development," and "management of service and staff" were the major areas of outsourcing. Outsourcing had been employed for 4-7 years by 45.5% of the hospitals and the proportion of the budget used for outsourcing was less than 20%. A need for an extension in outsourcing was agreed on by 76.5% of the hospitals. The multiple logistic regression analysis showed that both consumer satisfaction and security risk have an influence on hospital information system outsourcing. Outsourcing in hospital information systems is expected to increase just as in other industries. One primary facilitator to outsourcing in other industries is consumer satisfaction. We found that this was also a facilitator to outsourcing in hospital information systems. Security risk, which is usually considered an inhibitor to information technology outsourcing, was proven to be an inhibitor here as well. The results of this study may help hospital information systems establish a strategy and management plan for outsourcing.

  7. Facilitating superior chronic disease management through a knowledge-based systems development model.

    Science.gov (United States)

    Wickramasinghe, Nilmini S; Goldberg, Steve

    2008-01-01

    To date, the adoption and diffusion of technology-enabled solutions to deliver better healthcare has been slow. There are many reasons for this. One of the most significant is that the existing methodologies that are normally used in general for Information and Communications Technology (ICT) implementations tend to be less successful in a healthcare context. This paper describes a knowledge-based adaptive mapping to realisation methodology to traverse successfully from idea to realisation rapidly and without compromising rigour so that success ensues. It is discussed in connection with trying to implement superior ICT-enabled approaches to facilitate superior Chronic Disease Management (CDM).

  8. Cloning, Expression and Purification of the Recombinant HIV-1 Tat-Nef Fusion Protein in Prokaryotic Expression System

    Directory of Open Access Journals (Sweden)

    Somayeh Kadkhodayan

    2016-07-01

    Full Text Available Abstract Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa could act as a cell penetrating peptide (CPP. In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confirmed by western blot analysis and was purified using reverse staining method. Materials and Methods: In this experimental study, primarily, cloning of Tat-Nef fusion gene was done in pGEX6p2 expression vector. Then, the expression of Tat-Nef recombinat protein in E.coli BL21 (DE3 strain was performed by using IPTG inducer. The protein expression was confirmed by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant fusion protein was purified from gel using reverse staining method. Results: The results of PCR analysis and enzyme digestion showed a clear band of ~ 726 bp in agarose gel indicating the correct Tat-Nef fusion cloning in pGEX6p2 prokaryotic expression vector. In addition, a 54 kDa band of Tat-Nef on SDS-PAGE revealed Tat-Nef protein expression that western blot analysis using anti-Nef monoclonal antibody confirmed it. Conclusion: The purified Tat-Nef recombinant fusion protein will be used as an antigen for protein vaccine design against HIV infection.

  9. Laboratory observations and mathematical modeling of colloid-facilitated contaminant transport in chemically heterogeneous systems

    Science.gov (United States)

    Saiers, James E.

    2002-04-01

    In the work reported here, I examine colloid-facilitated solute transport through a natural porous medium that exhibits grain-scale heterogeneity in chemical composition. I conducted six column experiments using the contaminant hydroxyatrazine (HA) and either homogeneous clay-colloid suspensions or mixtures of clay colloids and dissolved organic matter. The transport of colloids composed of illite and montmorillonite, although not conservative, is only slightly affected by rate-limited mass transfer reactions with the oxide-coated aquifer material. Coinjection of HA with monomineralogic suspensions of illite colloids produces modest increases in effluent concentrations of HA relative to experiments performed with colloid-free influent solutions. Montmorillonite colloids have a much higher capacity to bind HA than illite colloids and substantially increase the transport rate of HA in the column experiments. Suwannee River humic acid (SRHA) does not strongly complex aqueous phase HA, but its addition to column influent mixtures that contain HA and montmorillonite suppresses HA adsorption to the clay colloids and promotes a decline in HA mobility. A model for colloid-facilitated transport, formulated to account for nonuniformity in the kinetics of colloid deposition and HA adsorption, closely describes HA transport in experiments performed with monomineralogic clay-colloid suspensions, heterogeneous suspensions made of both illite and montmorillonite, and mixtures of montmorillonite and SRHA.

  10. Basic Research Needs for Geosciences: Facilitating 21st Century Energy Systems

    Energy Technology Data Exchange (ETDEWEB)

    DePaolo, D. J.; Orr, F. M.; Benson, S. M.; Celia, M.; Felmy, A.; Nagy, K. L.; Fogg, G. E.; Snieder, R.; Davis, J.; Pruess, K.; Friedmann, J.; Peters, M.; Woodward, N. B.; Dobson, P.; Talamini, K.; Saarni, M.

    2007-06-01

    To identify research areas in geosciences, such as behavior of multiphase fluid-solid systems on a variety of scales, chemical migration processes in geologic media, characterization of geologic systems, and modeling and simulation of geologic systems, needed for improved energy systems.

  11. A Gateway MultiSite recombination cloning toolkit.

    Directory of Open Access Journals (Sweden)

    Lena K Petersen

    Full Text Available The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org.

  12. A Gateway MultiSite recombination cloning toolkit.

    Science.gov (United States)

    Petersen, Lena K; Stowers, R Steven

    2011-01-01

    The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org).

  13. A practical approach to T-cell receptor cloning and expression.

    Directory of Open Access Journals (Sweden)

    Sébastien Wälchli

    Full Text Available Although cloning and expression of T-cell Receptors (TcRs has been performed for almost two decades, these procedures are still challenging. For example, the use of T-cell clones that have undergone limited expansion as starting material to limit the loss of interesting TcRs, must be weighed against the introduction of mutations by excess PCR cycles. The recent interest in using specific TcRs for cancer immunotherapy has, however, increased the demand for practical and robust methods to rapidly clone and express TcRs. Two main technologies for TcR cloning have emerged; the use of a set of primers specifically annealing to all known TcR variable domains, and 5'-RACE amplification. We here present an improved 5'-RACE protocol that represents a fast and reliable way to identify a TcR from 10(5 cells only, making TcR cloning feasible without a priori knowledge of the variable domain sequence. We further present a detailed procedure for the subcloning of TcRα and β chains into an expression system. We show that a recombination-based cloning protocol facilitates simple and rapid transfer of the TcR transgene into different expression systems. The presented comprehensive method can be performed in any laboratory with standard equipment and with a limited amount of starting material. We finally exemplify the straightforwardness and reliability of our procedure by cloning and expressing several MART-1-specific TcRs and demonstrating their functionality.

  14. Practice and Preparation Time Facilitate System-Switching in Perceptual Categorization

    Directory of Open Access Journals (Sweden)

    Sébastien Hélie

    2017-11-01

    Full Text Available Mounting evidence suggests that category learning is achieved using different psychological and biological systems. While existing multiple-system theories and models of categorization may disagree about the number or nature of the different systems, all assume that people can switch between systems seamlessly. However, little empirical data has been collected to test this assumption, and recent available data suggest that system-switching is difficult. The main goal of this article is to identify factors influencing the proportion of participants who successfully learn to switch between procedural and declarative systems on a trial-by-trial basis. Specifically, we tested the effects of preparation time and practice, two factors that have been useful in task-switching, in a system-switching experiment. The results suggest that practice and preparation time can be beneficial to system-switching (as calculated by a higher proportion of switchers and lower switch costs, especially when they are jointly present. However, this improved system-switching comes at the cost of a larger button-switch interference when changing the location of the response buttons. The article concludes with a discussion of the implications of these findings for empirical research on system-switching and theoretical work on multiple-systems of category learning.

  15. A handheld support system to facilitate stereological measurements and mapping of branching structures

    DEFF Research Database (Denmark)

    Gardi, Jonathan Eyal; Wulfsohn, Dvoralai; Nyengaard, Jens Randel

    2007-01-01

    arterioles or for collection and optimizing sampling of crops for precision agriculture. Although the system has been designed specifically for sampling branching structures, it is sufficiently flexible to be used for other applications involving nested stereological designs. We describe the system...... specifications, software and Graphical User Interface (GUI) development, functionality and application of the handheld system using four examples: (1) sampling monkey lung bronchioles for estimation of diameter and wall thickness (2) sampling rat kidney for estimating number of arteries and arterioles...

  16. Cloning and joint measurements of incompatible components of spin

    International Nuclear Information System (INIS)

    Brougham, Thomas; Andersson, Erika; Barnett, Stephen M.

    2006-01-01

    A joint measurement of two observables is a simultaneous measurement of both quantities upon the same quantum system. When two quantum-mechanical observables do not commute, then a joint measurement of these observables cannot be accomplished directly by projective measurements alone. In this paper we shall discuss the use of quantum cloning to perform a joint measurement of two components of spin associated with a qubit system. We introduce cloning schemes which are optimal with respect to this task. The cloning schemes may be thought to work by cloning two components of spin onto their outputs. We compare the proposed cloning machines to existing cloners

  17. Facilitators and barriers to doing workplace mental health research: Case study of acute psychological trauma in a public transit system.

    Science.gov (United States)

    Links, Paul S; Bender, Ash; Eynan, Rahel; O'Grady, John; Shah, Ravi

    2016-03-10

    The Acute Psychological Trauma (APT) Study was a collaboration between an acute care hospital, a specialized multidisciplinary program designed to meet the mental health needs of injured workers, and a large urban public transit system. The overall purpose was to evaluate a Best Practices Intervention (BPI) for employees affected by acute psychological trauma compared to a Treatment as Usual (TAU) group. The specific purpose is to discuss facilitators and barriers that were recognized in implementing and carrying out mental health research in a workplace setting. Over the course of the APT study, a joint implementation committee was responsible for day-to-day study operations and made regular observations on the facilitators and barriers that arose throughout the study. The facilitators to this study included the longstanding relationships among the partners, increased recognition for the need of mental health research in the workplace, and the existence of a community advisory committee. The significant barriers to doing this study of mental health research in the workplace included differences in organizational culture, inconsistent union support, co-interventions, and stigma. Researchers and funding agencies need to be flexible and provide additional resources in order to overcome the barriers that can exist doing workplace mental health research.

  18. Construction of an infectious plasmid clone of Muscovy duck parvovirus by TA cloning and creation of a partially attenuated strain.

    Science.gov (United States)

    Yen, T-Y; Li, K-P; Ou, S-C; Shien, J-H; Lu, H-M; Chang, P-C

    2015-01-01

    Muscovy duck parvovirus (MDPV) infection is a highly contagious and fatal disease of Muscovy ducklings. The infectious clone methodology is a valuable tool to study the pathogenic mechanisms of viruses, but no infectious clone of MDPV is yet available. In this study, a plasmid clone containing the full-length genome of MDPV was constructed using the TA cloning methodology. This MDPV clone was found to be infectious after transfection of primary Muscovy duck embryo fibroblast cells and passage in embryonated Muscovy duck eggs. Site-directed mutagenesis showed that the K75N mutation in the VP1 protein of MDPV resulted in the partial attenuation of the virus. The availability of an MDPV infectious clone can facilitate investigation of the pathogenic mechanisms of MDPV and development of vaccines against diseases caused by MDPV.

  19. A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector.

    Science.gov (United States)

    Buj, Raquel; Iglesias, Noa; Planas, Anna M; Santalucía, Tomàs

    2013-08-20

    Valuable clone collections encoding the complete ORFeomes for some model organisms have been constructed following the completion of their genome sequencing projects. These libraries are based on Gateway cloning technology, which facilitates the study of protein function by simplifying the subcloning of open reading frames (ORF) into any suitable destination vector. The expression of proteins of interest as fusions with functional modules is a frequent approach in their initial functional characterization. A limited number of Gateway destination expression vectors allow the construction of fusion proteins from ORFeome-derived sequences, but they are restricted to the possibilities offered by their inbuilt functional modules and their pre-defined model organism-specificity. Thus, the availability of cloning systems that overcome these limitations would be highly advantageous. We present a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins based on the MultiSite Gateway cloning system. The fusion protein components are encoded in the three plasmids integral to the kit. These can recombine with any purposely-engineered destination vector that uses a heterologous promoter external to the Gateway cassette, leading to the in-frame cloning of an ORF of interest flanked by two functional modules. In contrast to previous systems, a third part becomes available for peptide-encoding as it no longer needs to contain a promoter, resulting in an increased number of possible fusion combinations. We have constructed the kit's component plasmids and demonstrate its functionality by providing proof-of-principle data on the expression of prototype fluorescent fusions in transiently-transfected cells. We have developed a toolkit for creating fusion proteins with customized N- and C-term modules from Gateway entry clones encoding ORFs of interest. Importantly, our method allows entry clones obtained from ORFeome collections to be used without prior

  20. The Power of 2: How an Apparently Irregular Numeration System Facilitates Mental Arithmetic

    Science.gov (United States)

    Bender, Andrea; Beller, Sieghard

    2017-01-01

    Mangarevan traditionally contained two numeration systems: a general one, which was highly regular, decimal, and extraordinarily extensive; and a specific one, which was restricted to specific objects, based on diverging counting units, and interspersed with binary steps. While most of these characteristics are shared by numeration systems in…

  1. Cloning-free CRISPR

    NARCIS (Netherlands)

    Arbab, Mandana; Srinivasan, Sharanya; Hashimoto, Tatsunori; Geijsen, Niels; Sherwood, Richard I.

    2015-01-01

    We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each

  2. SLiCE: a novel bacterial cell extract-based DNA cloning method

    OpenAIRE

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2012-01-01

    We describe a novel cloning method termed SLiCE (Seamless Ligation Cloning Extract) 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 bp) with or without flanking heterologous sequences and provides an effective strategy for directional s...

  3. Development of angle information system to facilitate the adjustment of needle-holding posture.

    Science.gov (United States)

    Cao, Yang; Liu, Li; Miura, Satoshi; Seki, Masaki; Kobayashi, Yo; Kawamura, Kazuya; Fujie, Masakatsu G

    2017-11-01

    Our purpose is to develop a system based on image processing methods that can inform users of the angular relationship between the needle and the forceps. The user thereby adjusts their needle grasping posture according to the angle information, which leads to an improvement in suturing accuracy. The system prototype consists of a camera and an image processing computer. The image captured by the camera is input to the computer, and then, the angular relationship between the forceps and needle is calculated via image processing. Then, the system informs the user of the calculated angular relationship between the needle and forceps in real time. To evaluate whether the system improves suturing accuracy, we invited 12 participants to enroll in an experiment based on a suturing task. The experimental results illustrate that the system allows participants to easily adjust the positional relationship between the needle and the forceps and that this adjusted angular relationship leads to higher suturing accuracy. Adjustment to holding the needle at a right angle before insertion has a critical effect on suturing quality. Therefore, we developed a system that informs the user of the angular relationship between the needle and the forceps. The results of the evaluation show that the system significantly improves the suturing accuracy of participants via informing them of the angle.

  4. Validation of a new control system for Elekta accelerators facilitating continuously variable dose rate

    DEFF Research Database (Denmark)

    Bertelsen, Anders; Lorenzen, Ebbe L; Brink, Carsten

    2011-01-01

    Elekta accelerators controlled by the current clinically used accelerator control system, Desktop 7.01 (D7), uses binned variable dose rate (BVDR) for volumetric modulated arc therapy (VMAT). The next version of the treatment control system (Integrity) supports continuously variable dose rate (CVDR......) as well as BVDR. Using CVDR opposed to BVDR for VMAT has the potential of reducing the treatment time but may lead to lower dosimetric accuracy due to faster moving accelerator parts. Using D7 and a test version of Integrity, differences in ability to control the accelerator, treatment efficiency......, and dosimetric accuracy between the two systems were investigated....

  5. Use of Telephonetics RESULTS computer-facilitated telephone system with automatic results upload.

    Science.gov (United States)

    Evans-Jones, J; Steedman, N; Newman, M; Jones, R; Milburn, A; O'Mahony, C

    2011-09-01

    Our genitourinary medicine service implemented an automated telephone results system in 2010. This system is used in other services but few are able to upload negative results automatically from laboratory software. The use of this system reduced unanswered calls to a telephone results line and also the number of calls that had to be connected to clinical staff. Patients have better access to their results in a timely manner, as per the requirements of the recent Standards for the Management of Sexually Transmitted Infections document.

  6. Short description of an alternative simplified method for screening recombinant clones within the "AdEasy-System" by Duplex-PCR

    Directory of Open Access Journals (Sweden)

    Kienle Peter

    2005-01-01

    Full Text Available Abstract Background Recombinant adenoviral vectors are highly efficient for in vitro and in vivo gene delivery. They can easily be produced in large numbers, transduce a wide variety of cell types and generate high levels of transgene expression. The AdEasy system is a widely used system for generating recombinant adenoviral vectors, which are created with a minimum of enzymatic manipulations and by employing homologous recombination in E. coli. In this paper we describe an alternative simplified method for screening recombinant DNA within the AdEasy system. This Duplex-PCR-method is independent of the transgene or insert and can be used for the complete AdEasy system. It is characterized by a simple standard protocol and the results can be obtained within a few hours. The PCR is run with two different primer sets. The primers KanaFor and KanaRev hybridizise with the Kanamycin resistence gene and AdFor and AdRev detect the adenoviral backbone. In case of recombinant clones, two diagnostic fragments with a size of 384 bp and 768 bp are generated. Results The practicability of this method was verified with three different transgenes: Cytosin Deaminase (AdCD, p53 (Adp53 and Granulocyte Macrophage Colony Stimulating Factor (AdGM-CSF. Recombinant clones are indicated by two diagnostic fragments and are then suitable for further processing. Conclusion In summary, the presented protocol allows fast detection of recombinants with an easy technique by minimizing the amount of necessary steps for generating a recombinant adenovirus. This method is time sparing and cost-effective.

  7. Developing a complex systems perspective for medical education to facilitate the integration of basic science and clinical medicine.

    Science.gov (United States)

    Aron, David C

    2017-04-01

    The purpose of medical education is to produce competent and capable professional practitioners who can combine the art and science of medicine. Moreover, this process must prepare individuals to practise in a field in which knowledge is increasing and the contexts in which that knowledge is applied are changing in unpredictable ways. The 'basic sciences' are important in the training of a physician. The goal of basic science training is to learn it in a way that the material can be applied in practice. Much effort has been expended to integrate basic science and clinical training, while adding many other topics to the medical curriculum. This effort has been challenging. The aims of the paper are (1) to propose a unifying conceptual framework that facilitates knowledge integration among all levels of living systems from cell to society and (2) illustrate the organizing principles with two examples of the framework in action - cybernetic systems (with feedback) and distributed robustness. Literature related to hierarchical and holarchical frameworks was reviewed. An organizing framework derived from living systems theory and spanning the range from molecular biology to health systems management was developed. The application of cybernetic systems to three levels (regulation of pancreatic beta cell production of insulin, physician adjustment of medication for glycaemic control and development and action of performance measures for diabetes care) was illustrated. Similarly distributed robustness was illustrated by the DNA damage response system and principles underlying patient safety. Each of the illustrated organizing principles offers a means to facilitate the weaving of basic science and clinical medicine throughout the course of study. The use of such an approach may promote systems thinking, which is a core competency for effective and capable medical practice. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  8. Health systems facilitators and barriers to the integration of HIV and chronic disease services: a systematic review.

    Science.gov (United States)

    Watt, Nicola; Sigfrid, Louise; Legido-Quigley, Helena; Hogarth, Sue; Maimaris, Will; Otero-García, Laura; Perel, Pablo; Buse, Kent; McKee, Martin; Piot, Peter; Balabanova, Dina

    2017-11-01

    Integration of services for patients with more than one diagnosed condition has intuitive appeal but it has been argued that the empirical evidence to support it is limited. We report the findings of a systematic review that sought to identify health system factors, extrinsic to the integration process, which either facilitated or hindered the integration of services for two common disorders, HIV and chronic non-communicable diseases. Findings were initially extracted and organized around a health system framework, followed by a thematic cross-cutting analysis and validation steps. Of the 150 articles included, 67% (n = 102) were from high-income countries. The articles explored integration with services for one or several chronic disorders, the most studied being alcohol or substance use disorders (47.7%), and mental health issues (29.5%). Four cross-cutting themes related to the health system were identified. The first and most common theme was the requirement for effective collaboration and coordination: formal and informal productive relationships throughout the system between providers and within teams, and between staff and patients. The second was the need for adequate and appropriately skilled and incentivized health workers-with the right expertise, training and operational support for the programme. The third was the need for supportive institutional structures and dedicated resources. The fourth was leadership in terms of political will, effective managerial oversight and organizational culture, indicating that actual implementation is as important as programme design. A fifth theme, outside the health system, but underpinning all aspects of the system operation, was that placing the patient at the centre of service delivery and responding holistically to their diverse needs. This was an important facilitator of integration. These findings confirm that integration processes in service delivery depend substantially for their success on characteristics of

  9. Critical Systems Thinking for the Facilitation of Conservation Planning in Philippine Coastal Management

    NARCIS (Netherlands)

    Larsen, R.K.

    2011-01-01

    In Critical Systems Thinking, the notion of boundary judgements represents a constructionist view on knowing as the bounding of components of reality into knowable objects. Cognitive boundary judgements determine observations (facts) and evaluations (values), which knowers appreciate and act in

  10. A Survey on Linked Data and the Social Web as facilitators for TEL recommender systems

    NARCIS (Netherlands)

    Dietze, Stefan; Drachsler, Hendrik; Daniela, Giordano

    2014-01-01

    Personalisation, adaptation and recommendation are central features of TEL environments. In this context, information retrieval techniques are applied as part of TEL recommender systems to filter and recommend learning resources or peer learners according to user preferences and requirements.

  11. A Novel Approach in Facilitating Aviation Emergency Procedure Learning and Recall through an Intuitive Pictorial System

    National Research Council Canada - National Science Library

    Estrada, Arthur; Keeley, Jennifer A; LeDuc, Patricia A; Bass, Julie M; Rouse, Tiffany N; Ramiccio, John G; Rowe, Terri L

    2007-01-01

    ...: the Intuitive Pictorial System (IPS). Descriptive and inferential statistics, along with correlation, were used to assess the study data, which determined statistically significant differences between the IPS and traditional training methods...

  12. Extension Activity Support System (EASY: A Web-Based Prototype for Facilitating Farm Management

    Directory of Open Access Journals (Sweden)

    Christopher Pettit

    2012-01-01

    Full Text Available In response to disparate advances in delivering spatial information to support agricultural extension activities, the Extension Activity Support System (EASY project was established to develop a vision statement and conceptual design for such a system based on a national needs assessment. Personnel from across Australia were consulted and a review of existing farm information/management software undertaken to ensure that any system that is eventually produced from the EASY vision will build on the strengths of existing efforts. This paper reports on the collaborative consultative process undertaken to create the EASY vision as well as the conceptual technical design and business models that could support a fully functional spatially enabled online system.

  13. Igloo: mobile learning system to facilitate and support learners and educators

    CSIR Research Space (South Africa)

    Ogunleye, S

    2009-12-01

    Full Text Available This paper describes the design, implementation and evaluation a mobile learning technology, namely IGLOO for mobile devices. It is intended to support educators as well as students in a learning environment. A working prototype system is discussed...

  14. JXTA: A Technology Facilitating Mobile P2P Health Management System

    OpenAIRE

    Rajkumar, Rajasekaran; Iyengar, Nallani Chackravatula Sriman Naraya

    2012-01-01

    Objectives Mobile JXTA (Juxtapose) gaining momentum and has attracted the interest of doctors and patients through P2P service that transmits messages. Audio and video can also be transmitted through JXTA. The use of mobile streaming mechanism with the support of mobile hospital management and healthcare system would enable better interaction between doctors, nurses, and the hospital. Experimental results demonstrate good performance in comparison with conventional systems. This study evaluat...

  15. Facilitators and impediments in the adoption and implementation of the HubSpot marketing automation system

    OpenAIRE

    Leinonen, Miira

    2017-01-01

    Marketing automation is growing popularity among companies but it has not yet received great academic interest. Therefore, neither marketing automation systems have been studied thoroughly. Other preceding systems in marketing and sales, such as customer relationship management (CRM) and sales force automation (SFA) have however been explored comprehensively. The goal of this research is to increase the understanding of successful adoption and implementation of a marketing automation syst...

  16. Software to Facilitate Remote Sensing Data Access for Disease Early Warning Systems.

    Science.gov (United States)

    Liu, Yi; Hu, Jiameng; Snell-Feikema, Isaiah; VanBemmel, Michael S; Lamsal, Aashis; Wimberly, Michael C

    2015-12-01

    Satellite remote sensing produces an abundance of environmental data that can be used in the study of human health. To support the development of early warning systems for mosquito-borne diseases, we developed an open-source, client based software application to enable the Epidemiological Applications of Spatial Technologies (EASTWeb). Two major design decisions were full automation of the discovery, retrieval and processing of remote sensing data from multiple sources, and making the system easily modifiable in response to changes in data availability and user needs. Key innovations that helped to achieve these goals were the implementation of a software framework for data downloading and the design of a scheduler that tracks the complex dependencies among multiple data processing tasks and makes the system resilient to external errors. EASTWeb has been successfully applied to support forecasting of West Nile virus outbreaks in the United States and malaria epidemics in the Ethiopian highlands.

  17. Rational Phosphorus Application Facilitates the Sustainability of the Wheat/Maize/Soybean Relay Strip Intercropping System.

    Directory of Open Access Journals (Sweden)

    Yuanxue Chen

    Full Text Available Wheat (Triticum aestivum L./maize (Zea mays L./soybean (Glycine max L. relay strip intercropping (W/M/S system is commonly used by the smallholders in the Southwest of China. However, little known is how to manage phosphorus (P to enhance P use efficiency of the W/M/S system and to mitigate P leaching that is a major source of pollution. Field experiments were carried out in 2011, 2012, and 2013 to test the impact of five P application rates on yield and P use efficiency of the W/M/S system. The study measured grain yield, shoot P uptake, apparent P recovery efficiency (PRE and soil P content. A linear-plateau model was used to determine the critical P rate that maximizes gains in the indexes of system productivity. The results show that increase in P application rates aggrandized shoot P uptake and crops yields at threshold rates of 70 and 71.5 kg P ha-1 respectively. With P application rates increasing, the W/M/S system decreased the PRE from 35.9% to 12.3% averaged over the three years. A rational P application rate, 72 kg P ha-1, or an appropriate soil Olsen-P level, 19.1 mg kg-1, drives the W/M/S system to maximize total grain yield while minimizing P surplus, as a result of the PRE up to 28.0%. We conclude that rational P application is an important approach for relay intercropping to produce high yield while mitigating P pollution and the rational P application-based integrated P fertilizer management is vital for sustainable intensification of agriculture in the Southwest of China.

  18. Validation of a new control system for Elekta accelerators facilitating continuously variable dose rate

    DEFF Research Database (Denmark)

    Bertelsen, Anders; Lorenzen, Ebbe L; Brink, Carsten

    2011-01-01

    Elekta accelerators controlled by the current clinically used accelerator control system, Desktop 7.01 (D7), uses binned variable dose rate (BVDR) for volumetric modulated arc therapy (VMAT). The next version of the treatment control system (Integrity) supports continuously variable dose rate (CVDR......) as well as BVDR. Using CVDR opposed to BVDR for VMAT has the potential of reducing the treatment time but may lead to lower dosimetric accuracy due to faster moving accelerator parts. Using D7 and a test version of Integrity, differences in ability to control the accelerator, treatment efficiency...

  19. What supports do health system organizations have in place to facilitate evidence-informed decision-making? A qualitative study.

    Science.gov (United States)

    Ellen, Moriah E; Léon, Gregory; Bouchard, Gisèle; Lavis, John N; Ouimet, Mathieu; Grimshaw, Jeremy M

    2013-08-06

    Decisions regarding health systems are sometimes made without the input of timely and reliable evidence, leading to less than optimal health outcomes. Healthcare organizations can implement tools and infrastructures to support the use of research evidence to inform decision-making. The purpose of this study was to profile the supports and instruments (i.e., programs, interventions, instruments or tools) that healthcare organizations currently have in place and which ones were perceived to facilitate evidence-informed decision-making. In-depth semi-structured telephone interviews were conducted with individuals in three different types of positions (i.e., a senior management team member, a library manager, and a 'knowledge broker') in three types of healthcare organizations (i.e., regional health authorities, hospitals and primary care practices) in two Canadian provinces (i.e., Ontario and Quebec). The interviews were taped, transcribed, and then analyzed thematically using NVivo 9 qualitative data analysis software. A total of 57 interviews were conducted in 25 organizations in Ontario and Quebec. The main findings suggest that, for the healthcare organizations that participated in this study, the following supports facilitate evidence-informed decision-making: facilitating roles that actively promote research use within the organization; establishing ties to researchers and opinion leaders outside the organization; a technical infrastructure that provides access to research evidence, such as databases; and provision and participation in training programs to enhance staff's capacity building. This study identified the need for having a receptive climate, which laid the foundation for the implementation of other tangible initiatives and supported the use of research in decision-making. This study adds to the literature on organizational efforts that can increase the use of research evidence in decision-making. Some of the identified supports may increase the use of

  20. The Game Embedded CALL System to Facilitate English Vocabulary Acquisition and Pronunciation

    Science.gov (United States)

    Young, Shelley Shwu-Ching; Wang, Yi-Hsuan

    2014-01-01

    The aim of this study is to make a new attempt to explore the potential of integrating game strategies with automatic speech recognition technologies to provide learners with individual opportunities for English pronunciation learning. The study developed the Game Embedded CALL (GeCALL) system with two activities for on-line speaking practice. For…

  1. The intrinsic features of Environmental Management Systems that facilitate adoption and encourage innovation in primary industries

    NARCIS (Netherlands)

    Carruthers, Genevieve; Vanclay, Frank

    2012-01-01

    This paper examines the theoretical underpinnings of the adoption of innovations, and applies this knowledge to the uptake of Environmental Management Systems (EMS) amongst Australian farmers. We examine the specific features of the EMS process that might encourage or inhibit EMS adoption. We also

  2. Facilitating Metacognitive Processes of Academic Genre-Based Writing Using an Online Writing System

    Science.gov (United States)

    Yeh, Hui-Chin

    2015-01-01

    Few studies have investigated how metacognitive processes foster the application of genre knowledge to students' academic writing. This is largely due to its internal and unobservable characteristics. To bridge this gap, an online writing system based on metacognition, involving the stages of planning, monitoring, evaluating, and revising, was…

  3. Effects of Storytelling to Facilitate EFL Speaking Using Web-Based Multimedia System

    Science.gov (United States)

    Hwang, Wu-Yuin; Shadiev, Rustam; Hsu, Jung-Lung; Huang, Yueh-Min; Hsu, Guo-Liang; Lin, Yi-Chun

    2016-01-01

    This study applied storytelling in the English as a foreign language (EFL) classroom in order to promote speaking skills. Students were asked to practice speaking EFL through producing individual and interactive stories with a Web-based multimedia system. We aimed to investigate an effectiveness of applying individual and interactive storytelling…

  4. Cultivation System Using Glass Beads Immersed in Liquid Medium Facilitates Studies of Streptomyces Differentiation

    Czech Academy of Sciences Publication Activity Database

    Nguyen, Liem Duy; Kalachová, Ladislava; Novotná, Jana; Holub, Martin; Kofroňová, Olga; Benada, Oldřich; Thompson, Ch.,J.; Weiser, Jaroslav

    2005-01-01

    Roč. 71, č. 6 (2005), s. 2848-2852 ISSN 0099-2240 R&D Projects: GA ČR(CZ) GA204/00/1252; GA ČR(CZ) GA204/03/1014 Institutional research plan: CEZ:AV0Z5020903 Keywords : cultivation system * streptomyces Subject RIV: EE - Microbiology , Virology Impact factor: 3.818, year: 2005

  5. Simplified cryopreservation of porcine cloned blastocysts

    DEFF Research Database (Denmark)

    Du, Yutao; Zhang, Yunhai; Li, Juan

    2007-01-01

    )â€"handmade cloning (HMC)â€"to establish a simplified and efficient cryopreservation system for porcine cloned embryos. In Experiment 1, zonae pellucidae of oocytes were partially digested with pronase, followed by centrifugation to polarize lipid particles. Ninety percent (173/192) oocytes were successfully......). Our results prove that porcine embryos produced from delipated oocytes by PA or HMC can be cryopreserved effectively by ultrarapid vitrification. Further experiments are required to assess the in vivo developmental competence of the cloned-vitrified embryos  ...

  6. Comparison of the miRNA profiles in HPV-positive and HPV-negative tonsillar tumors and a model system of human keratinocyte clones

    International Nuclear Information System (INIS)

    Vojtechova, Zuzana; Sabol, Ivan; Salakova, Martina; Smahelova, Jana; Zavadil, Jiri; Turek, Lubomir; Grega, Marek; Klozar, Jan; Prochazka, Bohumir; Tachezy, Ruth

    2016-01-01

    Better insights into the molecular changes involved in virus-associated and -independent head and neck cancer may advance our knowledge of HNC carcinogenesis and identify critical disease biomarkers. Here we aimed to characterize the expression profiles in a matched set of well-characterized HPV-dependent and HPV-independent tonsillar tumors and equivalent immortalized keratinocyte clones to define potential and clinically relevant biomarkers of HNC of different etiology. Fresh frozen tonsillar cancer tissues were analyzed together with non-malignant tonsillar tissues and compared with cervical tumors and normal cervical tissues. Furthermore, relative miRNAs abundance levels of primary and immortalized human keratinocyte clones were evaluated. The global quantitation of miRNA gene abundance was performed using a TaqMan Low Density Array system. The confirmation of differentially expressed miRNAs was performed on a set of formalin-fixed paraffin-embedded tumor samples enriched for the tumor cell fraction by macrodissection. We defined 46 upregulated and 31 downregulated miRNAs characteristic for the HPV-positive tonsillar tumors and 42 upregulated miRNAs and 42 downregulated miRNAs characteristic for HPV-independent tumors. In comparison with the expression profiles in cervical tumors, we defined miR-141-3p, miR-15b-5p, miR-200a-3p, miR-302c-3p, and miR-9-5p as specific for HPV induced malignancies. MiR-335-5p, miR-579-3p, and miR-126-5p were shared by the expression profiles of HPV-positive tonsillar tumors and of the HPV immortalized keratinocyte clones, whereas miR-328-3p, miR-34c-3p, and miR-885-5p were shared by the miRNA profiles of HPV-negative tonsillar tumors and the HPV-negative keratinocytes. We identified the miRNAs characteristic for HPV-induced tumors and tonsillar tumors of different etiology, and the results were compared with those of the model system. Our report presents the basis for further investigations leading to the identification of

  7. Triatoma infestans Calreticulin: Gene Cloning and Expression of a Main Domain That Interacts with the Host Complement System

    Science.gov (United States)

    Weinberger, Katherine; Collazo, Norberto; Aguillón, Juan Carlos; Molina, María Carmen; Rosas, Carlos; Peña, Jaime; Pizarro, Javier; Maldonado, Ismael; Cattan, Pedro E.; Apt, Werner; Ferreira, Arturo

    2017-01-01

    Triatoma infestans is an important hematophagous vector of Chagas disease, a neglected chronic illness affecting approximately 6 million people in Latin America. Hematophagous insects possess several molecules in their saliva that counteract host defensive responses. Calreticulin (CRT), a multifunctional protein secreted in saliva, contributes to the feeding process in some insects. Human CRT (HuCRT) and Trypanosoma cruzi CRT (TcCRT) inhibit the classical pathway of complement activation, mainly by interacting through their central S domain with complement component C1. In previous studies, we have detected CRT in salivary gland extracts from T. infestans. We have called this molecule TiCRT. Given that the S domain is responsible for C1 binding, we have tested its role in the classical pathway of complement activation in vertebrate blood. We have cloned and characterized the complete nucleotide sequence of CRT from T. infestans, and expressed its S domain. As expected, this S domain binds to human C1 and, as a consequence, it inhibits the classical pathway of complement, at its earliest stage of activation, namely the generation of C4b. Possibly, the presence of TiCRT in the salivary gland represents an evolutionary adaptation in hematophagous insects to control a potential activation of complement proteins, present in the massive blood meal that they ingest, with deleterious consequences at least on the anterior digestive tract of these insects. PMID:27895277

  8. Triatoma infestans Calreticulin: Gene Cloning and Expression of a Main Domain That Interacts with the Host Complement System.

    Science.gov (United States)

    Weinberger, Katherine; Collazo, Norberto; Aguillón, Juan Carlos; Molina, María Carmen; Rosas, Carlos; Peña, Jaime; Pizarro, Javier; Maldonado, Ismael; Cattan, Pedro E; Apt, Werner; Ferreira, Arturo

    2017-02-08

    Triatoma infestans is an important hematophagous vector of Chagas disease, a neglected chronic illness affecting approximately 6 million people in Latin America. Hematophagous insects possess several molecules in their saliva that counteract host defensive responses. Calreticulin (CRT), a multifunctional protein secreted in saliva, contributes to the feeding process in some insects. Human CRT (HuCRT) and Trypanosoma cruzi CRT (TcCRT) inhibit the classical pathway of complement activation, mainly by interacting through their central S domain with complement component C1. In previous studies, we have detected CRT in salivary gland extracts from T. infestans We have called this molecule TiCRT. Given that the S domain is responsible for C1 binding, we have tested its role in the classical pathway of complement activation in vertebrate blood. We have cloned and characterized the complete nucleotide sequence of CRT from T. infestans , and expressed its S domain. As expected, this S domain binds to human C1 and, as a consequence, it inhibits the classical pathway of complement, at its earliest stage of activation, namely the generation of C4b. Possibly, the presence of TiCRT in the salivary gland represents an evolutionary adaptation in hematophagous insects to control a potential activation of complement proteins, present in the massive blood meal that they ingest, with deleterious consequences at least on the anterior digestive tract of these insects. © The American Society of Tropical Medicine and Hygiene.

  9. Cloning of gene-encoded stem bromelain on system coming from Pichia pastoris as therapeutic protein candidate

    Science.gov (United States)

    Yusuf, Y.; Hidayati, W.

    2018-01-01

    The process of identifying bacterial recombination using PCR, and restriction, and then sequencing process was done after identifying the bacteria. This research aimed to get a yeast cell of Pichia pastoris which has an encoder gene of stem bromelain enzyme. The production of recombinant stem bromelain enzymes using yeast cells of P. pastoris can produce pure bromelain rod enzymes and have the same conformation with the enzyme’s conformation in pineapple plants. This recombinant stem bromelain enzyme can be used as a therapeutic protein in inflammatory, cancer and degenerative diseases. This study was an early stage of a step series to obtain bromelain rod protein derived from pineapple made with genetic engineering techniques. This research was started by isolating the RNA of pineapple stem which was continued with constructing cDNA using reserve transcriptase-PCR technique (RT-PCR), doing the amplification of bromelain enzyme encoder gene with PCR technique using a specific premiere couple which was designed. The process was continued by cloning into bacterium cells of Escherichia coli. A vector which brought the encoder gene of stem bromelain enzyme was inserted into the yeast cell of P. pastoris and was continued by identifying the yeast cell of P. pastoris which brought the encoder gene of stem bromelain enzyme. The research has not found enzyme gene of stem bromelain in yeast cell of P. pastoris yet. The next step is repeating the process by buying new reagent; RNase inhibitor, and buying liquid nitrogen.

  10. A digital, decentralized power station control system with bus-transmission facilitates the problem of backfitting

    International Nuclear Information System (INIS)

    Kaiser, G.E.; Schemmel, R.R.

    1985-01-01

    Current NPP control equipment technology is essentially characterized by the transmission of information in parallel using individual cables, and utilizes hardwired techniques for the processing of information. Progress in the area of semiconductor development characterized by micro-processors and LSI-circuits, has opened up new possibilities for the solution of the control tasks. The new power station control system PROCONTROL P utilizes these possibilities

  11. Fungal genome and mating system transitions facilitated by chromosomal translocations involving intercentromeric recombination.

    Directory of Open Access Journals (Sweden)

    Sheng Sun

    2017-08-01

    Full Text Available Species within the human pathogenic Cryptococcus species complex are major threats to public health, causing approximately 1 million annual infections globally. Cryptococcus amylolentus is the most closely known related species of the pathogenic Cryptococcus species complex, and it is non-pathogenic. Additionally, while pathogenic Cryptococcus species have bipolar mating systems with a single large mating type (MAT locus that represents a derived state in Basidiomycetes, C. amylolentus has a tetrapolar mating system with 2 MAT loci (P/R and HD located on different chromosomes. Thus, studying C. amylolentus will shed light on the transition from tetrapolar to bipolar mating systems in the pathogenic Cryptococcus species, as well as its possible link with the origin and evolution of pathogenesis. In this study, we sequenced, assembled, and annotated the genomes of 2 C. amylolentus isolates, CBS6039 and CBS6273, which are sexual and interfertile. Genome comparison between the 2 C. amylolentus isolates identified the boundaries and the complete gene contents of the P/R and HD MAT loci. Bioinformatic and chromatin immunoprecipitation sequencing (ChIP-seq analyses revealed that, similar to those of the pathogenic Cryptococcus species, C. amylolentus has regional centromeres (CENs that are enriched with species-specific transposable and repetitive DNA elements. Additionally, we found that while neither the P/R nor the HD locus is physically closely linked to its centromere in C. amylolentus, and the regions between the MAT loci and their respective centromeres show overall synteny between the 2 genomes, both MAT loci exhibit genetic linkage to their respective centromere during meiosis, suggesting the presence of recombinational suppressors and/or epistatic gene interactions in the MAT-CEN intervening regions. Furthermore, genomic comparisons between C. amylolentus and related pathogenic Cryptococcus species provide evidence that multiple chromosomal

  12. An integrative health information systems approach for facilitating strategic planning in hospitals.

    Science.gov (United States)

    Killingsworth, Brenda; Newkirk, Henry E; Seeman, Elaine

    2006-01-01

    This article presents a framework for developing strategic information systems (SISs) for hospitals. It proposes a SIS formulation process which incorporates complexity theory, strategic/organizational analysis theory, and conventional MIS development concepts. Within the formulation process, four dimensions of SIS are proposed as well as an implementation plan. A major contribution of this article is the development of a hospital SIS framework which permits an organization to fluidly respond to external, interorganizational, and intraorganizational influences. In addition, this article offers a checklist which managers can utilize in developing an SIS in health care.

  13. Tutor perceptions of the use of a reflective portfolio within a pastoral tutor system to facilitate undergraduate personal development planning.

    Science.gov (United States)

    Ellis, J S; Hobson, R S; Waterhouse, P J; Meechan, J G; Hogg, S D; Whitworth, J M; Thomason, J M

    2006-11-01

    All Higher Education Institutions in the UK are now required to provide transcripts of student activity and outcomes of summative assessments. In addition, the student should be able to reflect on their learning and plan their own development. This article reports on the staff evaluation of the use of a reflective portfolio facilitating the production of highly individualised personal development plans within an existing tutor system. A number of significant issues are highlighted; tutor systems adopting this approach must maintain flexibility for managing student crises when they arise, the difference between appraisal and assessment needs clear definition for both students and tutors, training in basic mentoring skills should be provided for all tutors, tutors should be aware of the difficulties many students experience with reflection and also be alert to the over reflective learner.

  14. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning

  15. Unified Approach to Universal Cloning and Phase-Covariant Cloning

    OpenAIRE

    Hu, Jia-Zhong; Yu, Zong-Wen; Wang, Xiang-Bin

    2008-01-01

    We analyze the problem of approximate quantum cloning when the quantum state is between two latitudes on the Bloch's sphere. We present an analytical formula for the optimized 1-to-2 cloning. The formula unifies the universal quantum cloning (UQCM) and the phase covariant quantum cloning.

  16. SIB-ESS-C - A Spatial Data Infrastructure to Facilitate Earth System Science in Siberia

    Science.gov (United States)

    Schmullius, C. C.; Gerlach, R.; Hese, S.

    2006-12-01

    The potential of Spatial Data Infrastructures (SDI) to share geoinformation and services over the Internet has been widely recognized and various initiatives (e.g. INSPIRE, GMES, GEOSS) are actively engaged to implement such systems on local, national or global level. Standards published by the Open Geospatial Consortium (OGC), the International Organization for Standardization (ISO) or the World Wide Web Consortium (W3C) provide the basis for distributed yet interoperable SDI's. Following the available guidelines and standards the Siberian Earth System Science Cluster (SIB-ESS-C) is being developed as a spatial data infrastructure for remote sensing product generation, data dissemination and scientific data analysis. This paper describes the concept of the Siberian Earth System Science Cluster. SIB-ESS-C emerged from the EU funded SIBERIA-II project (EVG2-2001-00008, 2002-2005) which was a joint Russian-European remote sensing project focusing on a 300 Million ha area in the central Siberian region. This area represents a significant part of the Earth's boreal biome which plays a critical role in global climate change in Northern Eurasia. The overall objective of the SIBERIA-II project was to demonstrate the viability of full carbon accounting including greenhouse gases (GHG) on a regional basis using state-of-the-art environmental methods and advanced remote sensing technologies. The tools and systems which have been employed include a selected yet spectrally and temporally diverse set of 15 Earth observation instruments on 8 satellites, detailed GIS databases and Dynamic Global Vegetation Models to account for fluxes between land and atmosphere. The data sets and value-added products created within the SIBERIA-II project will form the basic set of products for SIB-ESS-C. These products include regional maps of land cover, fire induced disturbances, phenology, snow depth, snow melt date, onset and duration of freeze and thaw, LAI and others. Since these products

  17. Facilitating the Easy Use of Earth Observation Data in Earth System Models through CyberConnector

    Science.gov (United States)

    Di, L.; Sun, Z.; Zhang, C.

    2017-12-01

    Earth system models (ESM) are an important tool used to understand the Earth system and predict its future states. On other hand, Earth observations (EO) provides the current state of the system. EO data are very useful in ESM initialization, verification, validation, and inter-comparison. However, EO data often cannot directly be consumed by ESMs because of the syntactic and semantic mismatches between EO products and ESM requirements. In order to remove the mismatches, scientists normally spend long time to customize EO data for ESM consumption. CyberConnector, a NSF EarthCube building block, is intended to automate the data customization so that scientists can be relieved from the laborious EO data customization. CyberConnector uses web-service-based geospatial processing models (GPM) as the mechanism to automatically customize the EO data into the right products in the right form needed by ESMs. It can support many different ESMs through its standard interfaces. It consists of seven modules: GPM designer, GPM binder, GPM runner, GPM monitor, resource register, order manager, and result display. In CyberConnector, EO data instances and GPMs are independent and loosely coupled. A modeler only needs to create a GPM in the GMP designer for EO data customization. Once the modeler specifies a study area, the designed GPM will be activated and take the temporal and spatial extents as constraints to search the data sources and customize the available EO data into the ESM-acceptable form. The execution of GMP is completely automatic. Currently CyberConnector has been fully developed. In order to validate the feasibility, flexibility, and ESM independence of CyberConnector, three ESMs from different geoscience disciplines, including the Cloud-Resolving Model (CRM), the Finite Volume Coastal Ocean Model (FVCOM), and the Community Multiscale Air Quality Model (CMAQ), have been experimented with CyberConnector through closely collaborating with modelers. In the experiment

  18. Application of geographic information systems and asset mapping to facilitate identification of colorectal cancer screening resources.

    Science.gov (United States)

    Gwede, Clement Kudzai; Ward, Beverly G; Luque, John S; Vadaparampil, Susan T; Rivers, Desiree; Martinez-Tyson, Dinorah; Noel-Thomas, Shalewa; Meade, Cathy D

    2010-01-01

    We sought to identify and map the geographic distribution of available colorectal cancer screening resources; following identification of this priority within a needs assessment of a local community-academic collaborative to reduce cancer health disparities in medically underserved communities. We used geographic information systems (GIS) and asset mapping tools to visually depict resources in the context of geography and a population of interest. We illustrate two examples, offer step-by-step directions for mapping, and discuss the challenges, lessons learned, and future directions for research and practice. Our positive asset driven, community-based approach illustrated the distribution of existing colonoscopy screening facilities and locations of populations and organizations who might use these resources. A need for additional affordable and accessible colonoscopy resources was identified. These transdisciplinary community mapping efforts highlight the benefit of innovative community-academic partnerships for addressing cancer health disparities by bolstering infrastructure and community capacity-building for increased access to colonoscopies.

  19. Integrated electrochemical treatment systems for facilitating the bioremediation of oil spill contaminated soil.

    Science.gov (United States)

    Cheng, Ying; Wang, Liang; Faustorilla, Vilma; Megharaj, Mallavarapu; Naidu, Ravi; Chen, Zuliang

    2017-05-01

    Bioremediation plays an important role in oil spill management and bio-electrochemical treatment systems are supposed to represent a new technology for both effective remediation and energy recovery. Diesel removal rate increased by four times in microbial fuel cells (MFCs) since the electrode served as an electron acceptor, and high power density (29.05 W m -3 ) at current density 72.38 A m -3 was achieved using diesel (v/v 1%) as the sole substrate. As revealed by Scanning electron microscope images, carbon fibres in the anode electrode were covered with biofilm and the bacterial colloids which build the link between carbon fibres and enhance electron transmission. Trace metabolites produced during the anaerobic biodegradation were identified by gas chromatography-mass spectrometry. These metabolites may act as emulsifying agents that benefit oil dispersion and play a vital role in bioremediation of oil spills in field applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Rapid cloning and purification of proteins: gateway vectors for protein purification by self-cleaving tags.

    Science.gov (United States)

    Gillies, Alison R; Hsii, Judy F; Oak, Seachol; Wood, David W

    2008-10-01

    We have combined Invitrogen's Gateway cloning technology with self-cleaving purification tags to generate a new system for rapid production of recombinant protein products. To accomplish this, we engineered our previously reported DeltaI-CM cleaving intein to include a Gateway cloning recognition sequence, and demonstrated that the resulting Gateway-competent intein is unaffected. This intein can therefore be used in several previously reported purification methods, while at the same time being compatible with Gateway cloning. We have incorporated this intein into a set of Gateway vectors, which include self-cleaving elastin-like polypeptide (ELP), chitin binding domain (CBD), phasin (polyhydroxybutyrate-binding), or maltose binding domain (MBD) tags. These vectors were verified by Gateway cloning of TEM-1 beta-lactamase and Escherichia coli catalase genes, and the expressed target proteins were purified using the four methods encoded on the vectors. The purification methods were unaffected by replacing the DeltaI-CM intein with the Gateway intein. It was observed that some purification methods were more appropriate for each target than others, suggesting utility of this technology for rapid process identification and optimization. The modular design of the Gateway system and intein purification method suggests that any tag and promoter can be trivially added to this system for the development of additional expression vectors. This technology could greatly facilitate process optimization, allowing several targets and methods to be tested in a high-throughput manner.

  1. Ground cover rice production system facilitates soil carbon and nitrogen stocks at regional scale

    Science.gov (United States)

    Liu, M.; Dannenmann, M.; Lin, S.; Saiz, G.; Yan, G.; Yao, Z.; Pelster, D.; Tao, H.; Sippel, S.; Tao, Y.; Zhang, Y.; Zheng, X.; Zuo, Q.; Butterbach-Bahl, K.

    2015-02-01

    Rice production is increasingly challenged by irrigation water scarcity, however covering paddy rice soils with films (ground cover rice production system: GCRPS) can significantly reduce water demand as well as overcome temperature limitations at the beginning of the vegetation period resulting in increased grain yields in colder regions of rice production with seasonal water shortages. It has been speculated that the increased soil aeration and temperature under GCRPS may result in losses of soil organic carbon and nitrogen stocks. Here we report on a regional scale experiment, conducted by sampling paired adjacent Paddy and GCRPS fields at 49 representative sites in the Shiyan region, which is typical for many mountainous areas across China. Parameters evaluated included soil C and N stocks, soil physical and chemical properties, potential carbon mineralization rates, fractions of soil organic carbon and stable carbon isotopic composition of plant leaves. Furthermore, root biomass was quantified at maximum tillering stage at one of our paired sites. Against expectations the study showed that: (1) GCRPS significantly increased soil organic C and N stocks 5-20 years following conversion of production systems, (2) there were no differences between GCRPS and Paddy in soil physical and chemical properties for the various soil depths with the exception of soil bulk density, (3) GCRPS had lower mineralization potential for soil organic C compared with Paddy over the incubation period, (4) GCRPS showed lower δ15N in the soils and plant leafs indicating less NH3 volatilization in GCRPS than in Paddy; and (5) GCRPS increased yields and root biomass in all soil layers down to 40 cm depth. Our results suggest that GCRPS is an innovative rice production technique that not only increases yields using less irrigation water, but that it also is environmentally beneficial due to increased soil C and N stocks at regional scale.

  2. Do Managers Clone Themselves?

    Science.gov (United States)

    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)

  3. Main: Clone Detail [KOME

    Lifescience Database Archive (English)

    Full Text Available Clone Detail Mapping Pseudomolecule data detail Detail information Mapping to the T...IGR japonica Pseudomolecules kome_mapping_pseudomolecule_data_detail.zip kome_mapping_pseudomolecule_data_detail ...

  4. Facilitating Semantic Interoperability Among Ocean Data Systems: ODIP-R2R Student Outcomes

    Science.gov (United States)

    Stocks, K. I.; Chen, Y.; Shepherd, A.; Chandler, C. L.; Dockery, N.; Elya, J. L.; Smith, S. R.; Ferreira, R.; Fu, L.; Arko, R. A.

    2014-12-01

    With informatics providing an increasingly important set of tools for geoscientists, it is critical to train the next generation of scientists in information and data techniques. The NSF-supported Rolling Deck to Repository (R2R) Program works with the academic fleet community to routinely document, assess, and preserve the underway sensor data from U.S. research vessels. The Ocean Data Interoperability Platform (ODIP) is an EU-US-Australian collaboration fostering interoperability among regional e-infrastructures through workshops and joint prototype development. The need to align terminology between systems is a common challenge across all of the ODIP prototypes. Five R2R students were supported to address aspects of semantic interoperability within ODIP. Developing a vocabulary matching service that links terms from different vocabularies with similar concept. The service implements Google Refine reconciliation service interface such that users can leverage Google Refine application as a friendly user interface while linking different vocabulary terms. Developing Resource Description Framework (RDF) resources that map Shipboard Automated Meteorological Oceanographic System (SAMOS) vocabularies to internationally served vocabularies. Each SAMOS vocabulary term (data parameter and quality control flag) will be described as an RDF resource page. These RDF resources allow for enhanced discoverability and retrieval of SAMOS data by enabling data searches based on parameter. Improving data retrieval and interoperability by exposing data and mapped vocabularies using Semantic Web technologies. We have collaborated with ODIP participating organizations in order to build a generalized data model that will be used to populate a SPARQL endpoint in order to provide expressive querying over our data files. Mapping local and regional vocabularies used by R2R to those used by ODIP partners. This work is described more fully in a companion poster. Making published Linked Data

  5. Emerging integrated nanoclay-facilitated drug delivery system for papillary thyroid cancer therapy

    Science.gov (United States)

    Zhang, Yi; Long, Mei; Huang, Peng; Yang, Huaming; Chang, Shi; Hu, Yuehua; Tang, Aidong; Mao, Linfeng

    2016-09-01

    Nanoclay can be incorporated into emerging dual functional drug delivery systems (DDSs) to promote efficiency in drug delivery and reduce the toxicity of doxorubicin (DOX) used for thyroid cancer treatment. This paper reports the expansion of the basal spacing of kaolinite nanoclay was expanded from 0.72 nm to 0.85 nm, which could provide sufficiently spacious site for hosting doxorubicin molecules and controlling the diffusion rate. A targeted design for papillary thyroid cancer cells was achieved by introducing KI, which is consumed by the sodium-iodide symporter (NIS). As indicated by MTT assays, confocal laser scanning microscopy and bio-TEM observations, methoxy-intercalated kaolinite (KaolinMeOH) exhibited negligible cytotoxicity against papillary thyroid cancer cells. By contrast, DOX-KaolinMeOH showed dose-dependent therapeutic effects in vitro, and KI@DOX-KaolinMeOH was found to act as a powerful targeted therapeutic drug. Furthermore, active and passive targeting strategies played a role in the accumulation of the drug molecules, as verified by an in vivo bio-distribution analysis.

  6. The intrinsic features of Environmental Management Systems that facilitate adoption and encourage innovation in primary industries.

    Science.gov (United States)

    Carruthers, Genevieve; Vanclay, Frank

    2012-11-15

    This paper examines the theoretical underpinnings of the adoption of innovations, and applies this knowledge to the uptake of Environmental Management Systems (EMS) amongst Australian farmers. We examine the specific features of the EMS process that might encourage or inhibit EMS adoption. We also consider elements of the EMS process to assess their utility in promoting adoption of various other innovations. We evaluate the EMS process in the light of two characteristics previously found to influence adoption of improved natural resource management practices - 'relative advantage' and 'trialability'. Drawing on literature, and our research and experience with farmers, we conclude that there are inherent features of EMS that promote the adoption of new practices, and that the elements of the EMS process actually create an on-going process of culture change as it is implemented. We believe that the EMS process offers significant advantages to farmers seeking to improve production whilst simultaneously meeting societal expectations for enhanced natural resource management. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Testosterone inhibits facilitating effects of parenting experience on parental behavior and the oxytocin neural system in mice.

    Science.gov (United States)

    Okabe, Shota; Kitano, Kanako; Nagasawa, Miho; Mogi, Kazutaka; Kikusui, Takefumi

    2013-06-13

    Parental behavior in mammals is facilitated by sensory experiences from infant, and by endocrine hormones. However, the interactions between these factors in the parental behavior of nonreproductive adults are not understood. We examined the interactive effects of gonadal hormones and the experience of repeated pup exposure on parental behavior in sexually naive mice. We also compared oxytocin (OT) expression levels in the paraventricular nucleus of the hypothalamus to behavioral outcomes. Clear sex differences were observed in retrieving tests; initial retrieving latency was shorter in females than in males, and 5-time pup exposure shortened retrieving latency in females only. Gonadectomy influenced neither initial retrieving latency nor pup sensitization in females. In contrast, gonadectomy shortened initial retrieving latency and caused pup sensitization in males. Estrogen implants given simultaneously with gonadectomy further shortened the initial retrieving latency in males, but pup sensitization was not affected and occurred in both sexes. In contrast, simultaneous testosterone implants impaired pup sensitization in both sexes. Similar to the results for responsiveness to pups, the number of OT neurons was increased by gonadectomy in males only. In comparison to gonadectomy only, OT neurons were decreased by simultaneous testosterone implants, but were not influenced by estrogen in either sex. Considering the parallel inhibitory effects of testosterone on both pup sensitization and number of OT neurons, we postulate that sex differences in parental responsiveness facilitated by repeated pup exposure were caused by an inhibitory effect of testosterone via the OT neural system in mice. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Distributed dendritic processing facilitates object detection: a computational analysis on the visual system of the fly.

    Directory of Open Access Journals (Sweden)

    Patrick Hennig

    Full Text Available BACKGROUND: Detecting objects is an important task when moving through a natural environment. Flies, for example, may land on salient objects or may avoid collisions with them. The neuronal ensemble of Figure Detection cells (FD-cells in the visual system of the fly is likely to be involved in controlling these behaviours, as these cells are more sensitive to objects than to extended background structures. Until now the computations in the presynaptic neuronal network of FD-cells and, in particular, the functional significance of the experimentally established distributed dendritic processing of excitatory and inhibitory inputs is not understood. METHODOLOGY/PRINCIPAL FINDINGS: We use model simulations to analyse the neuronal computations responsible for the preference of FD-cells for small objects. We employed a new modelling approach which allowed us to account for the spatial spread of electrical signals in the dendrites while avoiding detailed compartmental modelling. The models are based on available physiological and anatomical data. Three models were tested each implementing an inhibitory neural circuit, but differing by the spatial arrangement of the inhibitory interaction. Parameter optimisation with an evolutionary algorithm revealed that only distributed dendritic processing satisfies the constraints arising from electrophysiological experiments. In contrast to a direct dendro-dendritic inhibition of the FD-cell (Direct Distributed Inhibition model, an inhibition of its presynaptic retinotopic elements (Indirect Distributed Inhibition model requires smaller changes in input resistance in the inhibited neurons during visual stimulation. CONCLUSIONS/SIGNIFICANCE: Distributed dendritic inhibition of retinotopic elements as implemented in our Indirect Distributed Inhibition model is the most plausible wiring scheme for the neuronal circuit of FD-cells. This microcircuit is computationally similar to lateral inhibition between the

  9. Distributed dendritic processing facilitates object detection: a computational analysis on the visual system of the fly.

    Science.gov (United States)

    Hennig, Patrick; Möller, Ralf; Egelhaaf, Martin

    2008-08-28

    Detecting objects is an important task when moving through a natural environment. Flies, for example, may land on salient objects or may avoid collisions with them. The neuronal ensemble of Figure Detection cells (FD-cells) in the visual system of the fly is likely to be involved in controlling these behaviours, as these cells are more sensitive to objects than to extended background structures. Until now the computations in the presynaptic neuronal network of FD-cells and, in particular, the functional significance of the experimentally established distributed dendritic processing of excitatory and inhibitory inputs is not understood. We use model simulations to analyse the neuronal computations responsible for the preference of FD-cells for small objects. We employed a new modelling approach which allowed us to account for the spatial spread of electrical signals in the dendrites while avoiding detailed compartmental modelling. The models are based on available physiological and anatomical data. Three models were tested each implementing an inhibitory neural circuit, but differing by the spatial arrangement of the inhibitory interaction. Parameter optimisation with an evolutionary algorithm revealed that only distributed dendritic processing satisfies the constraints arising from electrophysiological experiments. In contrast to a direct dendro-dendritic inhibition of the FD-cell (Direct Distributed Inhibition model), an inhibition of its presynaptic retinotopic elements (Indirect Distributed Inhibition model) requires smaller changes in input resistance in the inhibited neurons during visual stimulation. Distributed dendritic inhibition of retinotopic elements as implemented in our Indirect Distributed Inhibition model is the most plausible wiring scheme for the neuronal circuit of FD-cells. This microcircuit is computationally similar to lateral inhibition between the retinotopic elements. Hence, distributed inhibition might be an alternative explanation of

  10. Systems biology studies of adult paragonimus lung flukes facilitate the identification of immunodominant parasite antigens.

    Directory of Open Access Journals (Sweden)

    Samantha N McNulty

    2014-10-01

    Full Text Available Paragonimiasis is a food-borne trematode infection acquired by eating raw or undercooked crustaceans. It is a major public health problem in the far East, but it also occurs in South Asia, Africa, and in the Americas. Paragonimus worms cause chronic lung disease with cough, fever and hemoptysis that can be confused with tuberculosis or other non-parasitic diseases. Treatment is straightforward, but diagnosis is often delayed due to a lack of reliable parasitological or serodiagnostic tests. Hence, the purpose of this study was to use a systems biology approach to identify key parasite proteins that may be useful for development of improved diagnostic tests.The transcriptome of adult Paragonimus kellicotti was sequenced with Illumina technology. Raw reads were pre-processed and assembled into 78,674 unique transcripts derived from 54,622 genetic loci, and 77,123 unique protein translations were predicted. A total of 2,555 predicted proteins (from 1,863 genetic loci were verified by mass spectrometric analysis of total worm homogenate, including 63 proteins lacking homology to previously characterized sequences. Parasite proteins encoded by 321 transcripts (227 genetic loci were reactive with antibodies from infected patients, as demonstrated by immunoaffinity purification and high-resolution liquid chromatography-mass spectrometry. Serodiagnostic candidates were prioritized based on several criteria, especially low conservation with proteins in other trematodes. Cysteine proteases, MFP6 proteins and myoglobins were abundant among the immunoreactive proteins, and these warrant further study as diagnostic candidates.The transcriptome, proteome and immunolome of adult P. kellicotti represent a major advance in the study of Paragonimus species. These data provide a powerful foundation for translational research to develop improved diagnostic tests. Similar integrated approaches may be useful for identifying novel targets for drugs and vaccines in the

  11. Systems biology studies of adult paragonimus lung flukes facilitate the identification of immunodominant parasite antigens.

    Science.gov (United States)

    McNulty, Samantha N; Fischer, Peter U; Townsend, R Reid; Curtis, Kurt C; Weil, Gary J; Mitreva, Makedonka

    2014-10-01

    Paragonimiasis is a food-borne trematode infection acquired by eating raw or undercooked crustaceans. It is a major public health problem in the far East, but it also occurs in South Asia, Africa, and in the Americas. Paragonimus worms cause chronic lung disease with cough, fever and hemoptysis that can be confused with tuberculosis or other non-parasitic diseases. Treatment is straightforward, but diagnosis is often delayed due to a lack of reliable parasitological or serodiagnostic tests. Hence, the purpose of this study was to use a systems biology approach to identify key parasite proteins that may be useful for development of improved diagnostic tests. The transcriptome of adult Paragonimus kellicotti was sequenced with Illumina technology. Raw reads were pre-processed and assembled into 78,674 unique transcripts derived from 54,622 genetic loci, and 77,123 unique protein translations were predicted. A total of 2,555 predicted proteins (from 1,863 genetic loci) were verified by mass spectrometric analysis of total worm homogenate, including 63 proteins lacking homology to previously characterized sequences. Parasite proteins encoded by 321 transcripts (227 genetic loci) were reactive with antibodies from infected patients, as demonstrated by immunoaffinity purification and high-resolution liquid chromatography-mass spectrometry. Serodiagnostic candidates were prioritized based on several criteria, especially low conservation with proteins in other trematodes. Cysteine proteases, MFP6 proteins and myoglobins were abundant among the immunoreactive proteins, and these warrant further study as diagnostic candidates. The transcriptome, proteome and immunolome of adult P. kellicotti represent a major advance in the study of Paragonimus species. These data provide a powerful foundation for translational research to develop improved diagnostic tests. Similar integrated approaches may be useful for identifying novel targets for drugs and vaccines in the future.

  12. A Citation Tracking System to Facilitate Sponsoring Institution Oversight of ACGME-Accredited Programs.

    Science.gov (United States)

    Long, Timothy R; Poe, John D; Zimmerman, Richard S; Rose, Steven H

    2012-12-01

    The Accreditation Council for Graduate Medical Education (ACGME) requires the graduate medical education committee and the designated institutional official to ensure that citations for noncompliance with the accreditation standards and institutional trends in citations are reviewed and corrected. To describe a citation tracking system (CTS) that uses Microsoft Office Access to efficiently catalogue, monitor, and document resolution of citations. The CTS was implemented in a sponsoring institution with oversight of 133 ACGME-accredited programs. The designated institutional official and the graduate medical education committee review all program letters of notification and enter citations into the CTS. A program-correction plan is required for each citation and is entered into the database. Open citations and action plans are reviewed by the graduate medical education committee and the designated institutional official on a quarterly basis, with decisions ranging from "closing" the citation to approving the action plan in process to requiring a new or modified action plan. Citation categories and subcategories are accessed on the ACGME website and entered into the CTS to identify trends. All 236 citations received since the 2006 Mayo School of Graduate Medical Education institutional site visit were entered into the CTS. On November 22, 2011, 26 of 236 citations (11%) were in active status with ongoing action plans, and 210 (89%) citations had been resolved and were closed. The CTS uses commercially available software to ensure citations are monitored and addressed and to simplify analysis of citation trends. The approach requires minimal staff time for data input and updates and can be performed without institutional information technology assistance.

  13. Transplantable living scaffolds comprised of micro-tissue engineered aligned astrocyte networks to facilitate central nervous system regeneration

    Science.gov (United States)

    Winter, Carla C.; Katiyar, Kritika S.; Hernandez, Nicole S.; Song, Yeri J.; Struzyna, Laura A.; Harris, James P.; Cullen, D. Kacy

    2017-01-01

    Neurotrauma, stroke, and neurodegenerative disease may result in widespread loss of neural cells as well as the complex interconnectivity necessary for proper central nervous system function, generally resulting in permanent functional deficits. Potential regenerative strategies involve the recruitment of endogenous neural stem cells and/or directed axonal regeneration through the use of tissue engineered “living scaffolds” built to mimic features of three-dimensional (3-D) in vivo migratory or guidance pathways. Accordingly, we devised a novel biomaterial encasement scheme using tubular hydrogel-collagen micro-columns that facilitated the self-assembly of seeded astrocytes into 3-D living scaffolds consisting of long, cable-like aligned astrocytic networks. Here, robust astrocyte alignment was achieved within a micro-column inner diameter (ID) of 180 μm or 300–350 μm but not 1.0 mm, suggesting that radius of curvature dictated the extent of alignment. Moreover, within small ID micro-columns, >70% of the astrocytes assumed a bi-polar morphology, versus ~10% in larger micro-columns or planar surfaces. Cell–cell interactions also influenced the aligned architecture, as extensive astrocyte-collagen contraction was achieved at high (9–12 × 105 cells/mL) but not lower (2–6 × 105 cells/mL) seeding densities. This high density micro-column seeding led to the formation of ultra-dense 3-D “bundles” of aligned bi-polar astrocytes within collagen measuring up to 150 μm in diameter yet extending to a remarkable length of over 2.5 cm. Importantly, co-seeded neurons extended neurites directly along the aligned astrocytic bundles, demonstrating permissive cues for neurite extension. These transplantable cable-like astrocytic networks structurally mimic the glial tube that guides neuronal progenitor migration in vivo along the rostral migratory stream, and therefore may be useful to guide progenitor cells to repopulate sites of widespread neurodegeneration

  14. Asymmetric quantum cloning machines

    International Nuclear Information System (INIS)

    Cerf, N.J.

    1998-01-01

    A family of asymmetric cloning machines for quantum bits and N-dimensional quantum states is introduced. These machines produce two approximate copies of a single quantum state that emerge from two distinct channels. In particular, an asymmetric Pauli cloning machine is defined that makes two imperfect copies of a quantum bit, while the overall input-to-output operation for each copy is a Pauli channel. A no-cloning inequality is derived, characterizing the impossibility of copying imposed by quantum mechanics. If p and p ' are the probabilities of the depolarizing channels associated with the two outputs, the domain in (√p,√p ' )-space located inside a particular ellipse representing close-to-perfect cloning is forbidden. This ellipse tends to a circle when copying an N-dimensional state with N→∞, which has a simple semi-classical interpretation. The symmetric Pauli cloning machines are then used to provide an upper bound on the quantum capacity of the Pauli channel of probabilities p x , p y and p z . The capacity is proven to be vanishing if (√p x , √p y , √p z ) lies outside an ellipsoid whose pole coincides with the depolarizing channel that underlies the universal cloning machine. Finally, the tradeoff between the quality of the two copies is shown to result from a complementarity akin to Heisenberg uncertainty principle. (author)

  15. The promise of dog cloning.

    Science.gov (United States)

    Oh, Hyun Ju; Ra, Kihae; Kim, Min Jung; Kim, Geon A; Setyawan, Erif Maha Nugraha; Lee, Seok Hee; Lee, Byeong Chun

    2017-01-01

    Dog cloning as a concept is no longer infeasible. Starting with Snuppy, the first cloned dog in the world, somatic cell nuclear transfer (SCNT) has been continuously developed and used for diverse purposes. In this article we summarise the current method for SCNT, the normality of cloned dogs and the application of dog cloning not only for personal reasons, but also for public purposes.

  16. Playing facilitator

    DEFF Research Database (Denmark)

    Houmøller, Ellen; Marchetti, Emanuela

    2015-01-01

    workshops based on two classic role-play games: The Silent Game (Brandt, 2006) and The Six Thinking Hats (de Bono, 1985). These games were created to support students in learning design thinking in groups and are assigned positive values in literature, hence we expected a smooth process. However, our......t: This paper presents reflections on the role of teachers as facilitators, in a context of role-play targeting learning of design thinking skills. Our study was conducted according to the method of visual ethnography. We acted as facilitators for 50 students through the yearly six-day competitive...... event called InnoEvent, addressed to students in the fields of multimedia and healthcare. Being interested in studying games and role-play as tools to support independent learning in the field of design thinking and team-building, following Dewey’s (1938) theory of learning experience, we ran two...

  17. Expert and novice facilitated modelling

    DEFF Research Database (Denmark)

    Tavella, Elena; Papadopoulos, Thanos

    2015-01-01

    the behaviour of one expert and two novice facilitators during a Viable System Model workshop. The findings suggest common facilitation patterns in the behaviour of experts and novices. This contrasts literature claiming that experts and novices behave and use their available knowledge differently......This paper provides an empirical study based on action research in which expert and novice facilitators in facilitated modelling workshops are compared. There is limited empirical research analysing the differences between expert and novice facilitators. Aiming to address this gap we study...... and facilitation strategies in contexts in which external, expert facilitation is not always possible are also discussed, and limitations of this study are provided....

  18. Vegetation pattern formation in semiarid systems induced by long-range competition in the absence of facilitation mechanisms.

    Science.gov (United States)

    Martinez-Garcia, Ricardo; Calabrese, Justin M.; Hernandez-Garcia, Emilio; Lopez, Cristobal

    2014-05-01

    Regular patterns and spatial organization of vegetation have been observed in many arid and semiarid ecosystems worldwide, covering a diverse range of plant taxa and soil types. A key common ingredient in these systems is that plant growth is severely limited by water availability, and thus plants likely compete strongly for water. The study of such patterns is especially interesting because their features may reveal much about the underlying physical and biological processes that generated them in addition to giving information on the characteristics of the ecosystem. It is possible, for instance, to infer their resilience against anthropogenic disturbances or climatic changes that could cause abrupt shifts in the system and lead it to a desert state. Therefore much research has focused on identifying the underlying mechanisms that can produce spatial patterning in water-limited systems (Klausmeier, 1999). They are believed to arise from the interplay between long-range competition and facilitation processes acting at smaller distances (Borgogno et al., 2009). This combination of mechanisms is justified by arguing that water percolates more readily through the soil in vegetated areas (short range), and that plants compete for water resources over greater distances via long lateral roots (long range). However, recent studies have shown that even in the limit of local facilitation patterns may still appear (Martinez-Garcia et al., 2013). In this work (Martinez-Garcia et al., 2013b), we show that, under rather general conditions, long-range competition alone is the minimal ingredient to shape gapped and stripped vegetation patterns typical of models that also account for facilitation in addition to competition. To this end we propose a simple, general model for the dynamics of vegetation, which includes only long-range competition between plants. Competition is introduced through a nonlocal term, where the kernel function quantifies the intensity of the interaction

  19. Grain legume-cereal intercropping: The practical application of diversity, competition and facilitation in arable and organic cropping systems

    DEFF Research Database (Denmark)

    Hauggaard-Nielsen, Henrik; Jørnsgaard, B.; Kinane, J.

    2008-01-01

    Intercropping is the simultaneous cultivation of more than one crop species on the same piece of land and is regarded as the practical application of basic ecological principles such as diversity, competition and facilitation. Field experiments were carried out on a sandy loam soil and a sandy soil......-270 g m-2). Nitrogen fixation was very constant in grain legume SC over species and location, varying from 13.2 to 15.8 g N m-2, being lowest in peas and highest in faba bean and lupin. The intercropped grain legumes increased the proportion of plant N derived from N2-fixation by on average 10...... was comparable; however, it tended to be the highest in sole cropped faba bean, lupin and unfertilized barley, where the application of urea to barley reduced the weed infestation by around 50%. Reduction in disease was observed in all IC systems compared to the corresponding SC, with a general disease reduction...

  20. Real-time photoacoustic and ultrasound dual-modality imaging system facilitated with graphics processing unit and code parallel optimization.

    Science.gov (United States)

    Yuan, Jie; Xu, Guan; Yu, Yao; Zhou, Yu; Carson, Paul L; Wang, Xueding; Liu, Xiaojun

    2013-08-01

    Photoacoustic tomography (PAT) offers structural and functional imaging of living biological tissue with highly sensitive optical absorption contrast and excellent spatial resolution comparable to medical ultrasound (US) imaging. We report the development of a fully integrated PAT and US dual-modality imaging system, which performs signal scanning, image reconstruction, and display for both photoacoustic (PA) and US imaging all in a truly real-time manner. The back-projection (BP) algorithm for PA image reconstruction is optimized to reduce the computational cost and facilitate parallel computation on a state of the art graphics processing unit (GPU) card. For the first time, PAT and US imaging of the same object can be conducted simultaneously and continuously, at a real-time frame rate, presently limited by the laser repetition rate of 10 Hz. Noninvasive PAT and US imaging of human peripheral joints in vivo were achieved, demonstrating the satisfactory image quality realized with this system. Another experiment, simultaneous PAT and US imaging of contrast agent flowing through an artificial vessel, was conducted to verify the performance of this system for imaging fast biological events. The GPU-based image reconstruction software code for this dual-modality system is open source and available for download from http://sourceforge.net/projects/patrealtime.

  1. High-dimensional quantum cloning and applications to quantum hacking.

    Science.gov (United States)

    Bouchard, Frédéric; Fickler, Robert; Boyd, Robert W; Karimi, Ebrahim

    2017-02-01

    Attempts at cloning a quantum system result in the introduction of imperfections in the state of the copies. This is a consequence of the no-cloning theorem, which is a fundamental law of quantum physics and the backbone of security for quantum communications. Although perfect copies are prohibited, a quantum state may be copied with maximal accuracy via various optimal cloning schemes. Optimal quantum cloning, which lies at the border of the physical limit imposed by the no-signaling theorem and the Heisenberg uncertainty principle, has been experimentally realized for low-dimensional photonic states. However, an increase in the dimensionality of quantum systems is greatly beneficial to quantum computation and communication protocols. Nonetheless, no experimental demonstration of optimal cloning machines has hitherto been shown for high-dimensional quantum systems. We perform optimal cloning of high-dimensional photonic states by means of the symmetrization method. We show the universality of our technique by conducting cloning of numerous arbitrary input states and fully characterize our cloning machine by performing quantum state tomography on cloned photons. In addition, a cloning attack on a Bennett and Brassard (BB84) quantum key distribution protocol is experimentally demonstrated to reveal the robustness of high-dimensional states in quantum cryptography.

  2. Ground-Based Global Navigation Satellite System (GNSS) GLONASS Broadcast Ephemeris Data (sub-hourly files) from NASA CDDIS - Cloned

    Data.gov (United States)

    National Aeronautics and Space Administration — This dataset consists of ground-based Global Navigation Satellite System (GNSS) GLObal NAvigation Satellite System (GLONASS) Broadcast Ephemeris Data (sub-hourly...

  3. The fibrinolytic system facilitates tumor cell migration across the blood-brain barrier in experimental melanoma brain metastasis

    International Nuclear Information System (INIS)

    Perides, George; Zhuge, Yuzheng; Lin, Tina; Stins, Monique F; Bronson, Roderick T; Wu, Julian K

    2006-01-01

    Patients with metastatic tumors to the brain have a very poor prognosis. Increased metastatic potential has been associated with the fibrinolytic system. We investigated the role of the fibrinolytic enzyme plasmin in tumor cell migration across brain endothelial cells and growth of brain metastases in an experimental metastatic melanoma model. Metastatic tumors to the brain were established by direct injection into the striatum or by intracarotid injection of B16F10 mouse melanoma cells in C57Bl mice. The role of plasminogen in the ability of human melanoma cells to cross a human blood-brain barrier model was studied on a transwell system. Wild type mice treated with the plasmin inhibitor epsilon-aminocaproic acid (EACA) and plg -/- mice developed smaller tumors and survived longer than untreated wild type mice. Tumors metastasized to the brain of wild type mice treated with EACA and plg -/- less efficiently than in untreated wild type mice. No difference was observed in the tumor growth in any of the three groups of mice. Human melanoma cells were able to cross the human blood-brain barrier model in a plasmin dependent manner. Plasmin facilitates the development of tumor metastasis to the brain. Inhibition of the fibrinolytic system could be considered as means to prevent tumor metastasis to the brain

  4. Time evolution of some quantum-mechanical systems. Wavefunction cloning in evolving rotating systems. Finite range boundary conditions for time dependent Schroedinger Equation

    International Nuclear Information System (INIS)

    Arvieu, R.; Carbonell, J.; Gignoux, C.; Mangin-Brinet, M.; Rozmej, P.

    1997-01-01

    The time evolution of coherent rotational wave packets associated to a diatomic molecule or to a deformed nucleus has been studied. Assuming a rigid body dynamics the J(J+1) law leads to a mechanism of cloning: the way function is divided into wave packets identical to the initial one at specific time. Applications are studied for a nuclear wave packed formed by Coulomb excitation. Exact boundary conditions at finite distance for the solution of the time-dependent Schroedinger equation are derived. A numerical scheme based on Crank-Nicholson method is proposed to illustrate its applicability in several examples. (authors)

  5. Integrating digital teaching-file systems with off-the-shelf presentation software to facilitate speaker-led conferences.

    Science.gov (United States)

    Frank, M S; Schultz, T; Dreyer, K

    2001-06-01

    Develop methods for automated transfer of images and associated text from a teaching-file repository into presentation material for speaker-led conferences. Our institution uses a Microsoft Windows (Microsoft Corp, Redmond, WA) software application to maintain a digital teaching-file database that can store and retrieve content in a case-centric fashion. Virtually any number of images can be stored with any given case. Cases and their associated images can be retrieved via a module that supports searches by American College of Radiology (ACR) code and by free-text Boolean queries on the history, findings, diagnosis, and discussion components of a case. In addition to the software system serving directly as an interactive teaching tool, the digital teaching file itself serves as an image repository and resource for attending radiologists who create their own presentations and lectures. To better support this use, software modules were developed for interprocess communication and automated creation of Powerpoint slides. These modules are fully integrated with the teaching-file software application. A single image or a set of selected images can be automatically made into individual slides with two mouse clicks. Images are automatically centered and optimally sized. A slide title is automatically rendered from the user's preference of the case history or diagnosis (stored with the case), or via the entry of freeform text. We describe the programming techniques that are used, as well as how several features of the operating system and Powerpoint itself can be integrated with a customized software application to facilitate this objective. The creation of presentation-ready Powerpoint slides is fully automated from within our teaching-file application, and the time required to create a presentation compared to the conventional method of manually seeking and inserting files from within Powerpoint itself, on a per-slide basis, is drastically reduced. The benefits are

  6. Potential for cloning dogs.

    Science.gov (United States)

    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.

  7. Effect of an emergency department-based electronic system for musculoskeletal consultation on facilitating care for common injuries.

    Science.gov (United States)

    Mears, Simon C; Pantle, Hardin A; Bessman, Edward S; Lifchez, Scott D

    2015-05-01

    Access to musculoskeletal consultation in the emergency department (ED) is a nationwide problem. In addition, consultation from a subspecialist may be delayed or may not be available, which can slow down the ED flow and reduce patient satisfaction. The purpose of this study was to review the 1-year results of a change in the authors' institutional practice to reduce subspecialty consultation for select musculoskeletal problems while still ensuring adequate patient follow-up in orthopedic or plastic surgery clinics for patients not seen by these services in the ED. The authors hypothesized that select injuries could be safely managed in the ED by using an electronic system to ensure appropriate follow-up care. Using Kaizen methodology, a multidisciplinary group (including ED staff, orthopedics, plastic surgery, pediatrics, nursing, radiology, therapy, and administration) met to improve care for select musculoskeletal injuries. A system was agreed on in which ED providers managed select musculoskeletal injuries without subspecialist consultation. Follow-up was organized using an electronic system, which facilitated communication between the ED staff and the secretarial staff of the subspecialist departments. Over a 1-year period, 150 patients were treated using this system. Charts and radiographs were reviewed for missed injuries. Radiographic review revealed 2 missed injuries. One patient had additional back pain and a lumbar spine fracture was found during the subspecialist follow-up visit; it was treated nonoperatively. Another patient appeared to have scapholunate widening on the injury radiograph that was not appreciated in the ED. Of the 150 patients, 51 were seen in follow-up by a subspecialist at the authors' institution. An electronic system to organize follow-up with a subspecialist allowed the ED providers to deliver safe and effective care for simple musculoskeletal injuries. Copyright 2015, SLACK Incorporated.

  8. Directly Transforming PCR-Amplified DNA Fragments into Plant Cells Is a Versatile System That Facilitates the Transient Expression Assay

    Science.gov (United States)

    Lu, Yuming; Chen, Xi; Wu, Yuxuan; Wang, Yanping; He, Yuqing; Wu, Yan

    2013-01-01

    A circular plasmid containing a gene coding sequence has been broadly used for studying gene regulation in cells. However, to accommodate a quick screen plasmid construction and preparation can be time consuming. Here we report a PCR amplified dsDNA fragments (PCR-fragments) based transient expression system (PCR-TES) for suiting in the study of gene regulation in plant cells. Instead of transforming plasmids into plant cells, transient expression of PCR-fragments can be applicable. The transformation efficiency and expression property of PCR-fragments are comparable to transformation using plasmids. We analyzed the transformation efficiency in PCR-TES at transcription and protein levels. Our results indicate that the PCR-TES is as versatile as the conventional transformation system using plasmid DNA. Through reconstituting PYR1-mediated ABA signaling pathway in Arabidopsis mesophyll protoplasts, we were not only validating the practicality of PCR-TES but also screening potential candidates of CDPK family members which might be involved in the ABA signaling. Moreover, we determined that phosphorylation of ABF2 by CPK4 could be mediated by ABA-induced PYR1 and ABI1, demonstrating a crucial role of CDPKs in the ABA signaling. In summary, PCR-TES can be applicable to facilitate analyzing gene regulation and for the screen of putative regulatory molecules at the high throughput level in plant cells. PMID:23468926

  9. Mutagenesis of the repeat regions of herpesviruses cloned as bacterial artificial chromosomes.

    Science.gov (United States)

    Zhao, Yuguang; Nair, Venugopal

    2010-01-01

    Cloning of infectious and pathogenic herpesvirus genomes in a bacterial artificial chromosome (BAC) vector greatly facilitates genetic manipulation of their genomes. BAC-based mutagenesis strategies of viruses can advance our understanding of the viral gene functions and determinants of pathogenicity, and can ultimately help to develop molecularly defined improved vaccines against virus diseases. Unlike the virus stocks, where continuous passage in tissue culture can lead to phenotypic alterations such as loss of virulence or immunogenicity, viral genomes can be stably maintained with high fidelity as BAC clones in bacteria. Thanks to the "RecA" or the inducible phage "lambda Red" homologous recombination systems and a variety of positive and negative selection strategies, viral genomes cloned as BAC can be efficiently manipulated in E. coli. All the manipulations, including DNA fragment deletion or insertion, point mutations, or even multiple modifications in repeat regions can be carried out accurately in E. coli, and the mutated DNA can be used directly to reconstitute mutant viruses in transfected host cells. Furthermore, using self-excision strategies, the non-viral bacterial replicon sequence can be excised automatically during virus reconstitution, thus generating recombinant viruses virtually identical to the wild-type parent viruses. Here, we describe the various technologies of manipulating the infectious BAC clones of a group E herpesvirus as an example through a combination of different approaches.

  10. Quantum Conditional Cloning of Continuous Variable Entangled States

    Science.gov (United States)

    Liu, K.; Gao, J. R.

    2014-12-01

    We extend the technique of conditional preparation to a quantum cloning machine, and present a protocol of 1 -> 2 conditional cloning of squeezed state and entanglement states. It is shown that the entanglement degree of the cloned entangled states can be well preserved even when the fidelity between the input and output states is beyond the limit of 4/9. This scheme is practicable since only the linear elements of beam splitters, homodyne detections, optical modulations and electrical trigger system, are involved.

  11. Hfq restructures RNA-IN and RNA-OUT and facilitates antisense pairing in the Tn10/IS10 system

    Science.gov (United States)

    Ross, Joseph A.; Ellis, Michael J.; Hossain, Shahan; Haniford, David B.

    2013-01-01

    Hfq functions in post-transcriptional gene regulation in a wide range of bacteria, usually by promoting base-pairing of mRNAs and trans-encoded sRNAs that share partial sequence complementarity. It is less clear if Hfq is required for pairing of cis-encoded RNAs (i.e., antisense RNAs) with their target mRNAs. In the current work, we have characterized the interactions between Escherichia coli Hfq and the components of the Tn10/IS10 antisense system, RNA-IN and RNA-OUT. We show that Hfq interacts with RNA-OUT through its proximal RNA-binding surface, as is typical for Hfq and trans-encoded sRNAs. In contrast, RNA-IN binds both proximal and distal RNA-binding surfaces in Hfq with a higher affinity for the latter, as is typical for mRNA interactions in canonical sRNA-mRNA pairs. Importantly, an amino acid substitution in Hfq that interferes with RNA binding to the proximal site negatively impacts RNA-IN:OUT pairing in vitro and suppresses the ability of Hfq to negatively regulate IS10 transposition in vivo. We also show that Hfq binding to RNA-IN and RNA-OUT alters secondary structure elements in both of these RNAs and speculate that this could be important in how Hfq facilitates RNA-IN:OUT pairing. Based on the results presented here, we suggest that Hfq could be involved in regulating RNA pairing in other antisense systems, including systems encoded by other transposable elements. PMID:23510801

  12. Facilitating medical information search using Google Glass connected to a content-based medical image retrieval system.

    Science.gov (United States)

    Widmer, Antoine; Schaer, Roger; Markonis, Dimitrios; Muller, Henning

    2014-01-01

    Wearable computing devices are starting to change the way users interact with computers and the Internet. Among them, Google Glass includes a small screen located in front of the right eye, a camera filming in front of the user and a small computing unit. Google Glass has the advantage to provide online services while allowing the user to perform tasks with his/her hands. These augmented glasses uncover many useful applications, also in the medical domain. For example, Google Glass can easily provide video conference between medical doctors to discuss a live case. Using these glasses can also facilitate medical information search by allowing the access of a large amount of annotated medical cases during a consultation in a non-disruptive fashion for medical staff. In this paper, we developed a Google Glass application able to take a photo and send it to a medical image retrieval system along with keywords in order to retrieve similar cases. As a preliminary assessment of the usability of the application, we tested the application under three conditions (images of the skin; printed CT scans and MRI images; and CT and MRI images acquired directly from an LCD screen) to explore whether using Google Glass affects the accuracy of the results returned by the medical image retrieval system. The preliminary results show that despite minor problems due to the relative stability of the Google Glass, images can be sent to and processed by the medical image retrieval system and similar images are returned to the user, potentially helping in the decision making process.

  13. Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon

    Directory of Open Access Journals (Sweden)

    Lawrence Mark L

    2010-07-01

    Full Text Available Abstract Background Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. Results We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. Conclusions This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.

  14. Use of Temporal Immersion Systems for the multiplication of nodal segments of Dioscorea alata L. in the ‘Pacala Duclos’ clone

    Directory of Open Access Journals (Sweden)

    Manuel Cabrera Jova

    2004-01-01

    Full Text Available In order to develop efficient propagation protocols, temporary immersion systems composed by two glass flask with 5 000 ml of capacity were used to multiply nodals segments from yam clone ‘‘Pacala Duclos’’. The working objectives for evaluation were: time and frequency of immersion, inoculum density, time and renewal volume of culture medium and multiplication stage length on propagation coefficient of nodal segments. Results permitted to define that using 10 minutes immersion time and immersion frequency each three and six hours, the highest increments in the multiplication coefficients (10.1 and 9.8 respectively, without significant differences between them were obtained. The most favorable result for the multiplication coefficient (10.8 was obtained when 50 explants per culture flask was inoculated. When the renewal was carried out 24 days after culture with 2 000 ml culture medium, the most advantageous result was shown for the multiplication coefficient, with 14.1. Subcultures were developed 49 days after culture because the highest coefficient multiplication (15.2 was noticed. A higher culture time did not influence in the multiplication coefficient significantly. Key words: culture medium, Dioscorea alata L., inoculum density, multiplication coefficient

  15. Applications of quantum cloning

    Science.gov (United States)

    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.

  16. Clip, connect, clone

    DEFF Research Database (Denmark)

    Fujima, Jun; Lunzer, Aran; Hornbæk, Kasper

    2010-01-01

    using three mechanisms: clipping of input and result elements from existing applications to form cells on a spreadsheet; connecting these cells using formulas, thus enabling result transfer between applications; and cloning cells so that multiple requests can be handled side by side. We demonstrate...

  17. Quantum cloning without signaling

    OpenAIRE

    Gisin, Nicolas

    1998-01-01

    Perfect Quantum Cloning Machines (QCM) would allow to use quantum nonlocality for arbitrary fast signaling. However perfect QCM cannot exist. We derive a bound on the fidelity of QCM compatible with the no-signaling constraint. This bound equals the fidelity of the Bu\\v{z}ek-Hillery QCM.

  18. Self-Cloning CRISPR

    NARCIS (Netherlands)

    Arbab, M.; Sherwood, R. I.

    2016-01-01

    CRISPR/Cas9-gene editing has emerged as a revolutionary technology to easily modify specific genomic loci by designing complementary sgRNA sequences and introducing these into cells along with Cas9. Self-cloning CRISPR/Cas9 (scCRISPR) uses a self-cleaving palindromic sgRNA plasmid (sgPal) that

  19. Prenatal music stimulation facilitates the postnatal functional development of the auditory as well as visual system in chicks (Gallus domesticus).

    Science.gov (United States)

    Roy, Saborni; Nag, Tapas C; Upadhyay, Ashish Datt; Mathur, Rashmi; Jain, Suman

    2014-03-01

    Rhythmic sound or music is known to improve cognition in animals and humans. We wanted to evaluate the effects of prenatal repetitive music stimulation on the remodelling of the auditory cortex and visual Wulst in chicks. Fertilized eggs (0 day) of white leghorn chicken (Gallus domesticus) during incubation were exposed either to music or no sound from embryonic day 10 until hatching. Auditory and visual perceptual learning and synaptic plasticity, as evident by synaptophysin and PSD-95 expression, were done at posthatch days (PH) 1, 2 and 3. The number of responders was significantly higher in the music stimulated group as compared to controls at PH1 in both auditory and visual preference tests. The stimulated chicks took significantly lesser time to enter and spent more time in the maternal area in both preference tests. A significantly higher expression of synaptophysin and PSD-95 was observed in the stimulated group in comparison to control at PH1-3 both in the auditory cortex and visual Wulst. A significant inter-hemispheric and gender-based difference in expression was also found in all groups. These results suggest facilitation of postnatal perceptual behaviour and synaptic plasticity in both auditory and visual systems following prenatal stimulation with complex rhythmic music.

  20. A system for household enumeration and re-identification in densely populated slums to facilitate community research, education, and advocacy.

    Directory of Open Access Journals (Sweden)

    Dana R Thomson

    Full Text Available We devised and implemented an innovative Location-Based Household Coding System (LBHCS appropriate to a densely populated informal settlement in Mumbai, India.LBHCS codes were designed to double as unique household identifiers and as walking directions; when an entire community is enumerated, LBHCS codes can be used to identify the number of households located per road (or lane segment. LBHCS was used in community-wide biometric, mental health, diarrheal disease, and water poverty studies. It also facilitated targeted health interventions by a research team of youth from Mumbai, including intensive door-to-door education of residents, targeted follow-up meetings, and a full census. In addition, LBHCS permitted rapid and low-cost preparation of GIS mapping of all households in the slum, and spatial summation and spatial analysis of survey data.LBHCS was an effective, easy-to-use, affordable approach to household enumeration and re-identification in a densely populated informal settlement where alternative satellite imagery and GPS technologies could not be used.

  1. Animal Cloning and Food Safety

    Science.gov (United States)

    ... For Consumers Home For Consumers Consumer Updates Animal Cloning and Food Safety Share Tweet Linkedin Pin it ... CVM could further evaluate the issue. FDA Studies Cloning For more than five years, CVM scientists studied ...

  2. The First Human Cloned Embryo.

    Science.gov (United States)

    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)

  3. [Nuclear transfer and therapeutic cloning].

    Science.gov (United States)

    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.

  4. The science and technology of farm animal cloning

    DEFF Research Database (Denmark)

    Gjerris, Mickey; Vajta, Gábor

    Details of the first mammal born after nuclear transfer cloning were published by Steen Malte Willadsen in 1986. In spite of its enormous scientific significance, this discovery failed to trigger much public concern, possibly because the donor cells were derived from pre-implantation stage embryos......, goats, horses, cats, etc. have been cloned with the somatic cell nuclear transfer technique. Although the technology still has relatively low success rates and there seems to be substantial problems with the welfare of some of the cloned animals, cloning is used both within basic research...... and the biomedical sector. The next step seems to be to implement cloning in the agricultural production system and several animals have been developed in this direction. This article reviews the current state of the art of farm animal cloning from a scientific and technological perspective, describes the animal...

  5. Human cloning. Fact or fiction

    International Nuclear Information System (INIS)

    Abushama, Mandy D.; Ahmed, Badreldeen I.

    2003-01-01

    Cloning is the production of one or more individual plants or animals that are genetically identical to other plant, animal or human. Scientists even demonstrated that they were able to clone frog tadpoles from frog embryonic cells using nuclear transfer.Many animals have been cloned from adult cells using nuclear transfer. Somatic cell nuclear transfer which refers to the transfer of the nucleous from a somatic cell to an egg cell. Article further deals with benefits and misuses of human cloning

  6. Complementation between two tospoviruses facilitates the systemic movement of a plant virus silencing suppressor in an otherwise restrictive host.

    Directory of Open Access Journals (Sweden)

    Sudeep Bag

    Full Text Available BACKGROUND: New viruses pathogenic to plants continue to emerge due to mutation, recombination, or reassortment among genomic segments among individual viruses. Tospoviruses cause significant economic damage to a wide range of crops in many parts of the world. The genetic or molecular basis of the continued emergence of new tospoviruses and new hosts is not well understood though it is generally accepted that reassortment and/or genetic complementation among the three genomic segments of individual viruses could be contributing to this variability since plants infected with more than one tospovirus are not uncommon in nature. METHODOLOGY/PRINCIPAL FINDINGS: Two distinct and economically important tospoviruses, Iris yellow spot virus (IYSV and Tomato spotted wilt virus (TSWV, were investigated for inter-virus interactions at the molecular level in dually-infected plants. Datura (Datura stramonium is a permissive host for TSWV, while it restricts the movement of IYSV to inoculated leaves. In plants infected with both viruses, however, TSWV facilitated the selective movement of the viral gene silencing suppressor (NSs gene of IYSV to the younger, uninoculated leaves. The small RNA expression profiles of IYSV and TSWV in single- and dually-infected datura plants showed that systemic leaves of dually-infected plants had reduced levels of TSWV N gene-specific small interfering RNAs (siRNAs. No TSWV NSs-specific siRNAs were detected either in the inoculated or systemic leaves of dually-infected datura plants indicating a more efficient suppression of host silencing machinery in the presence of NSs from both viruses as compared to the presence of only TSWV NSs. CONCLUSION/SIGNIFICANCE: Our study identifies a new role for the viral gene silencing suppressor in potentially modulating the biology and host range of viruses and underscores the importance of virally-coded suppressors of gene silencing in virus infection of plants. This is the first

  7. Complementation between Two Tospoviruses Facilitates the Systemic Movement of a Plant Virus Silencing Suppressor in an Otherwise Restrictive Host

    Science.gov (United States)

    Eid, Sahar; Pappu, Hanu R.

    2012-01-01

    Background New viruses pathogenic to plants continue to emerge due to mutation, recombination, or reassortment among genomic segments among individual viruses. Tospoviruses cause significant economic damage to a wide range of crops in many parts of the world. The genetic or molecular basis of the continued emergence of new tospoviruses and new hosts is not well understood though it is generally accepted that reassortment and/or genetic complementation among the three genomic segments of individual viruses could be contributing to this variability since plants infected with more than one tospovirus are not uncommon in nature. Methodology/Principal Findings Two distinct and economically important tospoviruses, Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV), were investigated for inter-virus interactions at the molecular level in dually-infected plants. Datura (Datura stramonium) is a permissive host for TSWV, while it restricts the movement of IYSV to inoculated leaves. In plants infected with both viruses, however, TSWV facilitated the selective movement of the viral gene silencing suppressor (NSs) gene of IYSV to the younger, uninoculated leaves. The small RNA expression profiles of IYSV and TSWV in single- and dually-infected datura plants showed that systemic leaves of dually-infected plants had reduced levels of TSWV N gene-specific small interfering RNAs (siRNAs). No TSWV NSs-specific siRNAs were detected either in the inoculated or systemic leaves of dually-infected datura plants indicating a more efficient suppression of host silencing machinery in the presence of NSs from both viruses as compared to the presence of only TSWV NSs. Conclusion/Significance Our study identifies a new role for the viral gene silencing suppressor in potentially modulating the biology and host range of viruses and underscores the importance of virally-coded suppressors of gene silencing in virus infection of plants. This is the first experimental evidence of

  8. A central support system can facilitate implementation and sustainability of a Classroom-based Undergraduate Research Experience (CURE) in Genomics.

    Science.gov (United States)

    Lopatto, David; Hauser, Charles; Jones, Christopher J; Paetkau, Don; Chandrasekaran, Vidya; Dunbar, David; MacKinnon, Christy; Stamm, Joyce; Alvarez, Consuelo; Barnard, Daron; Bedard, James E J; Bednarski, April E; Bhalla, Satish; Braverman, John M; Burg, Martin; Chung, Hui-Min; DeJong, Randall J; DiAngelo, Justin R; Du, Chunguang; Eckdahl, Todd T; Emerson, Julia; Frary, Amy; Frohlich, Donald; Goodman, Anya L; Gosser, Yuying; Govind, Shubha; Haberman, Adam; Hark, Amy T; Hoogewerf, Arlene; Johnson, Diana; Kadlec, Lisa; Kaehler, Marian; Key, S Catherine Silver; Kokan, Nighat P; Kopp, Olga R; Kuleck, Gary A; Lopilato, Jane; Martinez-Cruzado, Juan C; McNeil, Gerard; Mel, Stephanie; Nagengast, Alexis; Overvoorde, Paul J; Parrish, Susan; Preuss, Mary L; Reed, Laura D; Regisford, E Gloria; Revie, Dennis; Robic, Srebrenka; Roecklien-Canfield, Jennifer A; Rosenwald, Anne G; Rubin, Michael R; Saville, Kenneth; Schroeder, Stephanie; Sharif, Karim A; Shaw, Mary; Skuse, Gary; Smith, Christopher D; Smith, Mary; Smith, Sheryl T; Spana, Eric P; Spratt, Mary; Sreenivasan, Aparna; Thompson, Jeffrey S; Wawersik, Matthew; Wolyniak, Michael J; Youngblom, James; Zhou, Leming; Buhler, Jeremy; Mardis, Elaine; Leung, Wilson; Shaffer, Christopher D; Threlfall, Jennifer; Elgin, Sarah C R

    2014-01-01

    In their 2012 report, the President's Council of Advisors on Science and Technology advocated "replacing standard science laboratory courses with discovery-based research courses"-a challenging proposition that presents practical and pedagogical difficulties. In this paper, we describe our collective experiences working with the Genomics Education Partnership, a nationwide faculty consortium that aims to provide undergraduates with a research experience in genomics through a scheduled course (a classroom-based undergraduate research experience, or CURE). We examine the common barriers encountered in implementing a CURE, program elements of most value to faculty, ways in which a shared core support system can help, and the incentives for and rewards of establishing a CURE on our diverse campuses. While some of the barriers and rewards are specific to a research project utilizing a genomics approach, other lessons learned should be broadly applicable. We find that a central system that supports a shared investigation can mitigate some shortfalls in campus infrastructure (such as time for new curriculum development, availability of IT services) and provides collegial support for change. Our findings should be useful for designing similar supportive programs to facilitate change in the way we teach science for undergraduates. © 2014 D. Lopatto et al. CBE—Life Sciences Education © 2014 The American Society for Cell Biology. This article is distributed by The American Society for Cell Biology under license from the author(s). It is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  9. 4-Fragment Gateway cloning format for MosSCI-compatible vectors integrating Promoterome and 3'UTRome libraries of Caenorhabditis elegans.

    Science.gov (United States)

    Kogame, Toshiaki

    2015-01-01

    The technique of Mos1-mediated Single Copy Insertion (MosSCI) now has become the essential technique which facilitates transgenic experiments for Caenohabditis elegans (C. elegans). Gateway system which is adopted to MosSCI-compatible vectors offers an advantage of simultaneous cloning with entry vectors cloned in the Gateway system format. On the other hand, the format for MosSCI-compatible vectors restricts flexibility in designing the vectors to only 3-fragment integration. Thus, construct of complex transgene such as the expression vector for translational gene fusion is tedious work even with Gateway system. We have developed the new recombination format called LeGaSCI (Library-enhanced Gateway for MosSCI) to expand the conventional 3-fragment to 4-fragment format which still retains the capacity to accept Promoterome and 3'UTRome libraries of C. elegans. In the new recombination format, 2 different Gateway format were combined. Cloning reaction for the tissue-specific expression vector of GFP-tagged protein with 3'UTR successfully occurred without any expected insertion, deletion or frame-shift mutation. Moreover, The MosSCI transgenic line was successfully generated with the construct. Collectively, we established the new Gateway system format which allows us to assemble 4-fragment insertion with the widest variety of entry clone vectors from C. elegans libraries.

  10. Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

    Directory of Open Access Journals (Sweden)

    Samiha Sioud

    2009-01-01

    Full Text Available Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E. coli ET12567 (pUZ8002, as a conjugal donor, carrying the integrative plasmid pSET152. For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces sp. TN 58, the integration was in single copy at the attB site. Plasmid pSET152 was inherited every time for all analysed Streptomyces sp. US 24 and Streptomyces sp. TN 58 exconjugants under nonselective conditions. The growth, morphological differentiation, and active molecules production of all studied pSET152 integrated exconjugants were identical to those of wild type strains. Consequently, conjugal transfer using pSET152 integration system is a suitable means of genes transfer and expression for both studied strains. To validate the above gene transfer system, the glucose isomerase gene (xylA from Streptomyces sp. SK was expressed in strain Streptomyces sp. TN 58. Obtained results indicated that heterologous glucose isomerase could be expressed and folded effectively. Glucose isomerase activity of the constructed TN 58 recombinant strain is of about eighteenfold higher than that of the Streptomyces sp. SK strain. Such results are certainly of importance due to the potential use of improved strains in biotechnological process for the production of high-fructose syrup from starch.

  11. Three concepts of cloning in human beings.

    Science.gov (United States)

    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.

  12. Construction of a full-length infectious bacterial artificial chromosome clone of duck enteritis virus vaccine strain

    Science.gov (United States)

    2013-01-01

    Background Duck enteritis virus (DEV) is the causative agent of duck viral enteritis, which causes an acute, contagious and lethal disease of many species of waterfowl within the order Anseriformes. In recent years, two laboratories have reported on the successful construction of DEV infectious clones in viral vectors to express exogenous genes. The clones obtained were either created with deletion of viral genes and based on highly virulent strains or were constructed using a traditional overlapping fosmid DNA system. Here, we report the construction of a full-length infectious clone of DEV vaccine strain that was cloned into a bacterial artificial chromosome (BAC). Methods A mini-F vector as a BAC that allows the maintenance of large circular DNA in E. coli was introduced into the intergenic region between UL15B and UL18 of a DEV vaccine strain by homologous recombination in chicken embryoblasts (CEFs). Then, the full-length DEV clone pDEV-vac was obtained by electroporating circular viral replication intermediates containing the mini-F sequence into E. coli DH10B and identified by enzyme digestion and sequencing. The infectivity of the pDEV-vac was validated by DEV reconstitution from CEFs transfected with pDEV-vac. The reconstructed virus without mini-F vector sequence was also rescued by co-transfecting the Cre recombinase expression plasmid pCAGGS-NLS/Cre and pDEV-vac into CEF cultures. Finally, the in vitro growth properties and immunoprotection capacity in ducks of the reconstructed viruses were also determined and compared with the parental virus. Results The full genome of the DEV vaccine strain was successfully cloned into the BAC, and this BAC clone was infectious. The in vitro growth properties of these reconstructions were very similar to parental DEV, and ducks immunized with these viruses acquired protection against virulent DEV challenge. Conclusions DEV vaccine virus was cloned as an infectious bacterial artificial chromosome maintaining full

  13. Energetic funnel facilitates facilitated diffusion.

    Science.gov (United States)

    Cencini, Massimo; Pigolotti, Simone

    2018-01-25

    Transcription factors (TFs) are able to associate to their binding sites on DNA faster than the physical limit posed by diffusion. Such high association rates can be achieved by alternating between three-dimensional diffusion and one-dimensional sliding along the DNA chain, a mechanism-dubbed facilitated diffusion. By studying a collection of TF binding sites of Escherichia coli from the RegulonDB database and of Bacillus subtilis from DBTBS, we reveal a funnel in the binding energy landscape around the target sequences. We show that such a funnel is linked to the presence of gradients of AT in the base composition of the DNA region around the binding sites. An extensive computational study of the stochastic sliding process along the energetic landscapes obtained from the database shows that the funnel can significantly enhance the probability of TFs to find their target sequences when sliding in their proximity. We demonstrate that this enhancement leads to a speed-up of the association process. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  14. Mammalian cloning: advances and limitations.

    Science.gov (United States)

    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.

  15. Optimally cloned binary coherent states

    Science.gov (United States)

    Müller, C. R.; Leuchs, G.; Marquardt, Ch.; Andersen, U. L.

    2017-10-01

    Binary coherent state alphabets can be represented in a two-dimensional Hilbert space. We capitalize this formal connection between the otherwise distinct domains of qubits and continuous variable states to map binary phase-shift keyed coherent states onto the Bloch sphere and to derive their quantum-optimal clones. We analyze the Wigner function and the cumulants of the clones, and we conclude that optimal cloning of binary coherent states requires a nonlinearity above second order. We propose several practical and near-optimal cloning schemes and compare their cloning fidelity to the optimal cloner.

  16. A novel two-component signaling system facilitates uropathogenic Escherichia coli's ability to exploit abundant host metabolites.

    Directory of Open Access Journals (Sweden)

    Wentong Cai

    Full Text Available Two-component signaling systems (TCSs are major mechanisms by which bacteria adapt to environmental conditions. It follows then that TCSs would play important roles in the adaptation of pathogenic bacteria to host environments. However, no pathogen-associated TCS has been identified in uropathogenic Escherichia coli (UPEC. Here, we identified a novel TCS, which we termed KguS/KguR (KguS: α-ketoglutarate utilization sensor; KguR: α-ketoglutarate utilization regulator in UPEC CFT073, a strain isolated from human pyelonephritis. kguS/kguR was strongly associated with UPEC but was found only rarely among other E. coli including commensal and intestinal pathogenic strains. An in vivo competition assay in a mouse UTI model showed that deletion of kguS/kguR in UPEC CFT073 resulted in a significant reduction in its colonization of the bladders and kidneys of mice, suggesting that KguS/KguR contributed to UPEC fitness in vivo. Comparative proteomics identified the target gene products of KguS/KguR, and sequence analysis showed that TCS KguS/KguR and its targeted-genes, c5032 to c5039, are encoded on a genomic island, which is not present in intestinal pathogenic E. coli. Expression of the target genes was induced by α-ketoglutarate (α-KG. These genes were further shown to be involved in utilization of α-KG as a sole carbon source under anaerobic conditions. KguS/KguR contributed to the regulation of the target genes with the direct regulation by KguR verified using an electrophoretic mobility shift assay. In addition, oxygen deficiency positively modulated expression of kguS/kguR and its target genes. Taken altogether, this study describes the first UPEC-associated TCS that functions in controlling the utilization of α-ketoglutarate in vivo thereby facilitating UPEC adaptation to life inside the urinary tract.

  17. LinkHub: a Semantic Web system that facilitates cross-database queries and information retrieval in proteomics

    Directory of Open Access Journals (Sweden)

    Cheung Kei-Hoi

    2007-05-01

    Full Text Available Abstract Background A key abstraction in representing proteomics knowledge is the notion of unique identifiers for individual entities (e.g. proteins and the massive graph of relationships among them. These relationships are sometimes simple (e.g. synonyms but are often more complex (e.g. one-to-many relationships in protein family membership. Results We have built a software system called LinkHub using Semantic Web RDF that manages the graph of identifier relationships and allows exploration with a variety of interfaces. For efficiency, we also provide relational-database access and translation between the relational and RDF versions. LinkHub is practically useful in creating small, local hubs on common topics and then connecting these to major portals in a federated architecture; we have used LinkHub to establish such a relationship between UniProt and the North East Structural Genomics Consortium. LinkHub also facilitates queries and access to information and documents related to identifiers spread across multiple databases, acting as "connecting glue" between different identifier spaces. We demonstrate this with example queries discovering "interologs" of yeast protein interactions in the worm and exploring the relationship between gene essentiality and pseudogene content. We also show how "protein family based" retrieval of documents can be achieved. LinkHub is available at hub.gersteinlab.org and hub.nesg.org with supplement, database models and full-source code. Conclusion LinkHub leverages Semantic Web standards-based integrated data to provide novel information retrieval to identifier-related documents through relational graph queries, simplifies and manages connections to major hubs such as UniProt, and provides useful interactive and query interfaces for exploring the integrated data.

  18. The implementation of a quality system in the Dutch GP specialty training: barriers and facilitators; a qualitative study.

    Science.gov (United States)

    Buwalda, Nienke; Braspenning, Jozé; van Roosmalen, Sanne; van Dijk, Nynke; Visser, Mechteld

    2017-07-21

    Quality assurance programs in medical education are introduced to gain insight into the quality of such programs and to trigger improvements. Although of utmost importance, research on the implementation of such programs is scarce. The Dutch General Practice (GP) specialty training institutes used an implementation strategy to implement a quality system (QS), and we aimed to study the success of this strategy and to learn about additional facilitators and barriers. Seventeen structured interviews were conducted with the directors and quality coordinators (QCs) of the eight Dutch GP training institutes. A five-stage process model of implementation was used to structure these interviews and analyze the data. Two researchers analyzed the data with a framework approach. The strategy supported the institutes in implementing the QS. However, after the introduction of the QS, staff experienced the QS as demanding, although they noticed almost no concrete short-term results. Moreover, they experienced difficulties in integrating the QS into their local situation. Collectively working with the QS and following common deadlines did create a sense of commitment towards each other that appeared to be a true stimulus to the introduction of the QS. The implementation strategy focused mainly on the introduction of the QS in the GP specialty training, and it was, as such, rather successful. An important barrier concerned the acceptance of the QS and the integration of the QS into local structures, which suggests that there is a need for guidance on the translation of the QS to local contexts. All in all, we recommend more focus on the benefits of a QS.

  19. Probabilistic cloning of equidistant states

    International Nuclear Information System (INIS)

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

    2010-01-01

    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.

  20. Ethical issues in livestock cloning.

    Science.gov (United States)

    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.

  1. Facilitating leadership team communication

    OpenAIRE

    Hedman, Eerika

    2015-01-01

    The purpose of this study is to understand and describe how to facilitate competent communication in leadership teamwork. Grounded in the premises of social constructionism and informed by such theoretical frameworks as coordinated management of meaning theory (CMM), dialogic organization development (OD), systemic-constructionist leadership, communication competence, and reflexivity, this study seeks to produce further insights into understanding leadership team communicati...

  2. Biodiversity versus cloning

    International Nuclear Information System (INIS)

    Jaramillo T, Jose Hernan

    1998-01-01

    The announcement has been made on the cloning of mice in these days and he doesn't stop to miss, because the world lives a stage where conscience of the protection is creating that should be given to the biodiversity. It is known that alone we won't subsist and the protection of the means and all that contains that environment is of vital importance for the man. But it is also known that the vegetables and animal transgenic that they come to multiply the species have appeared that we prepare. The transgenic has been altered genetically, for substitution of one or more genes of other species, inclusive human genes. This represents an improvement compared with the investigations that gave origin to the cloning animal. But it is necessary to notice that to it you arrived through the cloning. This year 28 million hectares have been sowed in cultivations of transgenic seeds and there is around 700 bovine transgenic whose milk contains a necessary protein in the treatment of the man's illnesses

  3. Malaria in Kakuma refugee camp, Turkana, Kenya: facilitation of Anopheles arabiensis vector populations by installed water distribution and catchment systems

    Directory of Open Access Journals (Sweden)

    Cetron Martin S

    2011-06-01

    to infection with P. falciparum and showed a hyperendemic age-prevalence profile, in an area with otherwise low risk of malaria given prevailing climate. Transmission was sustained by A. arabiensis, whose populations were facilitated by installation of man-made water distribution and catchment systems.

  4. Cloning and immunoreactivity of the 5-HT1Mac and 5-HT2Mac receptors in the central nervous system of the freshwater prawn Macrobrachium rosenbergii

    Science.gov (United States)

    Vázquez-Acevedo, Nietzell; Reyes-Colón, Dalynés; Ruíz-Rodríguez, Eduardo A.; Rivera, Nilsa M.; Rosenthal, Joshua; Kohn, Andrea B.; Moroz, Leonid L.; Sosa, María A.

    2009-01-01

    Biogenic amines are implicated in several mental disorders, many of which involve social interactions. Simple model systems, such as crustaceans, are often more amenable than vertebrates for studying mechanisms underlying behaviors. Although various cellular responses of biogenic amines have been characterized in crustaceans, the mechanisms linking these molecules to behavior remain largely unknown. Observed effects of serotonin receptor agonists and antagonists in abdomen posture, escape responses, and fighting have led to the suggestion that biogenic amine receptors may play a role in modulating interactive behaviors. As a first step in understanding this potential role of such receptors, we have cloned and fully sequenced two serotonin receptors, 5-HT1Mac and 5-HT2Mac, from the CNS of the freshwater prawn Macrobrachium rosenbergii, and have mapped their CNS immunohistochemical distribution. 5-HT1Mac was found primarily on the membranes of subsets of cells in all CNS ganglia, in fibers that traverse all CNS regions, and in the cytoplasm of a small number of cells in the brain, circum- and subesophageal ganglia (SEG), most of which also appear to contain dopamine. The pattern of 5-HT2Mac immunoreactivity was found to differ significantly, being found mostly in the central neuropil area of all ganglia, in glomeruli of the brain’s olfactory lobes, and in the cytoplasm of a small number of neurons in the SEG, thoracic and some abdominal ganglia. The observed differences in terms of localization, distribution within cells, and intensity of immunoreactive staining throughout the prawn’s CNS suggest that these receptors are likely to play different roles. PMID:19184976

  5. Cloning and biochemical analysis of β-glucoside utilization (bgl) operon without phosphotransferase system in Pectobacterium carotovorum subsp. carotovorum LY34.

    Science.gov (United States)

    An, Chang Long; Kim, Min Keun; Kang, Tae Ho; Kim, Jungho; Kim, Hoon; Yun, Han Dae

    2012-09-06

    β-Glucosidases are widespread in bacteria and involved in the metabolism of various carbohydrate substrates. Studying of β-glucoside utilization (bgl) operons on operon of Pectobacterium carotovorum subsp. carotovorum LY34 (Pcc LY34) will help us understanding how β-glucoside utilization (bgl) operon can cooperate with other systems in bacterium caused soft-rot disease. Pcc LY34 causes soft-rot disease in plants and expresses multiple enzymatic forms of β-glucosidases. To fully explore the β-glucoside utilization system in Pcc LY34, we have isolated a bgl operon from a genomic library for screening of β-glucosidase activities. Sequence analysis of a 3050bp cloned DNA fragment (accession number AY870655) shows two open reading frames (bglY and bglK) that are predicted to encode proteins of 474 and 278 amino acid residues, respectively. Pair wise similarity analysis suggests BglY is a beta-glucosidase (a member of glycosyl hydrolase family 1) and BglK is a transcriptional antiterminator protein. bglY promoter region contains an inverted repeat sequence similar to transcriptional terminator. Different from other four β-glucoside utilization operons of Pcc LY34 strain, BglY contains signal peptide sequences as extracellular β-glucosidase. Comparisons of five β-glucoside utilization operons of Pcc LY34 strain showed that bglYK operon does not have phosphotransferase system domain which are responsible for sugar transportation. BglY shares 33-44% identity with other four β-glucosidases of Pcc LY34 strain. Enzyme assay showed that purified BglY enzyme hydrolyzed salicin, arbutin, pNPG, and MUG, and exhibited maximal activity at pH 7.0 and 40°C. This activity was enhanced Mg(2+). Site-directed mutagenesis revealed E166 and E371 are critical of BglY's β-glucosidase activity. Copyright © 2012 Elsevier GmbH. All rights reserved.

  6. Ethical issues in animal cloning.

    Science.gov (United States)

    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.

  7. The KnownLeaf literature curation system captures knowledge about Arabidopsis leaf growth and development and facilitates integrated data mining

    NARCIS (Netherlands)

    Szakonyi, D.; Landeghem, van S.; Baerenfaller, K.; Baeyens, L.; Blomme, J.; Casanova-Saéz, R.; Bodt, De S.; Esteve-Bruna, D.; Fiorani, F.; Gonzalez, N.; Grønlund, J.; Immink, R.G.H.; Jover-Gil, S.; Kuwabara, A.; Muñoz-Nortes, T.; Dijk, van A.D.J.; Wilson-Sánchez, D.; Buchanan-Wollaston, V.; Angenent, G.C.; Peer, Van de Y.; Inzé, D.; Micol, J.L.; Gruissem, W.; Walsh, S.; Hilson, P.

    2015-01-01

    The information that connects genotypes and phenotypes is essentially embedded in research articles written in natural language. To facilitate access to this knowledge, we constructed a framework for the curation of the scientific literature studying the molecular mechanisms that control leaf growth

  8. Local cloning of CAT states

    International Nuclear Information System (INIS)

    Rahaman, Ramij

    2011-01-01

    In this Letter we analyze the (im)possibility of the exact cloning of orthogonal three-qubit CAT states under local operation and classical communication (LOCC) with the help of a restricted entangled state. We also classify the three-qubit CAT states that can (not) be cloned under LOCC restrictions and extend the results to the n-qubit case. -- Highlights: → We analyze the (im)possibility of exact cloning of orthogonal CAT states under LOCC. → We also classify the set of CAT states that can(not) be cloned by LOCC. → No set of orthogonal CAT states can be cloned by LOCC with help of similar CAT state. → Any two orthogonal n-qubit GHZ-states can be cloned by LOCC with help of a GHZ state.

  9. Lessons learned from cloning dogs.

    Science.gov (United States)

    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.

  10. An online practice and educational networking system for technical skills: learning experience in expert facilitated vs. independent learning communities.

    Science.gov (United States)

    Rojas, David; Cheung, Jeffrey J H; Weber, Bryce; Kapralos, Bill; Carnahan, Heather; Bägli, Darius J; Dubrowski, Adam

    2012-01-01

    This study explored the activities of trainees learning technical skills using an educational networking tool with and without expert facilitation. Medical students (participants) were video-recorded practicing suturing and knot tying techniques and the resulting videos were uploaded to an educational networking site. Participants were then divided into two groups (one group containing an expert facilitator while the other group did not) and encouraged to comment on the videos within their group. We monitored the number of logins and comments posted and all participants completed an exit survey. There were no differences between the activities the two groups (p = 0.387). We conclude that the presence of an expert within collaborative Internet environments in not necessary to promote interactivity amongst the learners.

  11. Therapeutic cloning: The ethical limits

    International Nuclear Information System (INIS)

    Whittaker, Peter A.

    2005-01-01

    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. A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector

    Science.gov (United States)

    2013-01-01

    Background Valuable clone collections encoding the complete ORFeomes for some model organisms have been constructed following the completion of their genome sequencing projects. These libraries are based on Gateway cloning technology, which facilitates the study of protein function by simplifying the subcloning of open reading frames (ORF) into any suitable destination vector. The expression of proteins of interest as fusions with functional modules is a frequent approach in their initial functional characterization. A limited number of Gateway destination expression vectors allow the construction of fusion proteins from ORFeome-derived sequences, but they are restricted to the possibilities offered by their inbuilt functional modules and their pre-defined model organism-specificity. Thus, the availability of cloning systems that overcome these limitations would be highly advantageous. Results We present a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins based on the MultiSite Gateway cloning system. The fusion protein components are encoded in the three plasmids integral to the kit. These can recombine with any purposely-engineered destination vector that uses a heterologous promoter external to the Gateway cassette, leading to the in-frame cloning of an ORF of interest flanked by two functional modules. In contrast to previous systems, a third part becomes available for peptide-encoding as it no longer needs to contain a promoter, resulting in an increased number of possible fusion combinations. We have constructed the kit’s component plasmids and demonstrate its functionality by providing proof-of-principle data on the expression of prototype fluorescent fusions in transiently-transfected cells. Conclusions We have developed a toolkit for creating fusion proteins with customized N- and C-term modules from Gateway entry clones encoding ORFs of interest. Importantly, our method allows entry clones obtained from ORFeome

  13. Understanding Systems Change in Early Implementation of Housing First in Canadian Communities: An Examination of Facilitators/Barriers, Training/Technical Assistance, and Points of Leverage.

    Science.gov (United States)

    Worton, S Kathleen; Hasford, Julian; Macnaughton, Eric; Nelson, Geoffrey; MacLeod, Timothy; Tsemberis, Sam; Stergiopoulos, Vicky; Goering, Paula; Aubry, Tim; Distasio, Jino; Richter, Tim

    2018-03-01

    We present interim findings of a cross-site case study of an initiative to expand Housing First (HF) in Canada through training and technical assistance (TTA). HF is an evidence-based practice designed to end chronic homelessness for consumers of mental health services. We draw upon concepts from implementation science and systems change theory to examine how early implementation occurs within a system. Case studies examining HF early implementation were conducted in six Canadian communities receiving HF TTA. The primary data are field notes gathered over 1.5 years and evaluations from site-specific training events (k = 5, n = 302) and regional network training events (k = 4, n = 276). We report findings related to: (a) the facilitators of and barriers to early implementation, (b) the influence of TTA on early implementation, and (c) the "levers" used to facilitate broader systems change. Systems change theory enabled us to understand how various "levers" created opportunities for change within the communities, including establishing system boundaries, understanding how systems components can function as causes of or solutions to a problem, and assessing and changing systems interactions. We conclude by arguing that systems theory adds value to existing implementation science frameworks and can be helpful in future research on the implementation of evidence-based practices such as HF which is a complex community intervention. Implications for community psychology are discussed. © Society for Community Research and Action 2017.

  14. Characterization of forage and extrusa clones dwarf elephant grass under rotational stocking

    Directory of Open Access Journals (Sweden)

    Tatiana Pires Pereira

    2014-10-01

    Full Text Available The objective of this trial was to evaluate the behavior ingestive of crossbred heifers and chemical characteristics of the material from two clones of dwarf elephant grass (BRS Kurumi ‘and CNPGL 01/03/00 submitted to different management strategies through sampling of forage (whole plant extrusa and manual hand plucking. The experiment was conducted at Embrapa Dairy Cattle, Coronel Pacheco, MG. We used a completely randomized design with factorial (2x2x2 with three replications. The treatments consisted of two clones of elephant grass (BRS Kurumi ‘and CNPGL 01/03/00, two light interception at the entrance of the animals (90 and 95% and two heights of post-grazing residue (30 and 50 cm with three replications. The chemical analysis showed that the methodology manual grazing simulation enables an acceptable estimate of the forage selected by grazing animals and the sampling of the whole plant is not selected by the animal diet. To harvest extrusa rate evaluation and mass bit, fractions and chemical composition of the plant of the ingested material was taken. Characteristics, structural and nutritional value of clone BRS ‘Kurumi’ facilitated greater forage intake by the animal, suggesting its use in grazing systems.

  15. Cloning-independent plasmid construction for genetic studies in streptococci.

    Science.gov (United States)

    Xie, Zhoujie; Qi, Fengxia; Merritt, Justin

    2013-08-01

    Shuttle plasmids are among the few routinely utilized tools in the Streptococcus mutans genetic system that still require the use of classical cloning methodologies and intermediate hosts for genetic manipulation. Accordingly, it typically requires considerably less time and effort to introduce mutations onto the S. mutans chromosome than it does to construct shuttle vectors for expressing genes in trans. Occasionally, shuttle vector constructs also exhibit toxicity in Escherichia coli, which prevents their proper assembly. To circumvent these limitations, we modified a prolonged overlap extension PCR (POE-PCR) protocol to facilitate direct plasmid assembly in S. mutans. Using solely PCR, we created the reporter vector pZX7, which contains a single minimal streptococcal replication origin and harbors a spectinomycin resistance cassette and the gusA gene encoding β-glucuronidase. We compared the efficiency of pZX7 assembly using multiple strains of S. mutans and were able to obtain from 5 × 10³ to 2 × 10⁵ CFU/μg PCR product. Likewise, we used pZX7 to further demonstrate that Streptococcus sanguinis and Streptococcus gordonii are also excellent hosts for cloning-independent plasmid assembly, which suggests that this system is likely to function in numerous other streptococci. Consequently, it should be possible to completely forgo the use of E. coli-Streptococcus shuttle vectors in many streptococcal species, thereby decreasing the time and effort required to assemble constructs and eliminating any toxicity issues associated with intermediate hosts. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. [The discrete horror of cloning].

    Science.gov (United States)

    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.

  17. Local cloning of CAT states

    Science.gov (United States)

    Rahaman, Ramij

    2011-06-01

    In this Letter we analyze the (im)possibility of the exact cloning of orthogonal three-qubit CAT states under local operation and classical communication (LOCC) with the help of a restricted entangled state. We also classify the three-qubit CAT states that can (not) be cloned under LOCC restrictions and extend the results to the n-qubit case.

  18. [Scientific ethics of human cloning].

    Science.gov (United States)

    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.

  19. Extremal asymmetric universal cloning machines

    Science.gov (United States)

    Jiang, Mingming; Yu, Sixia

    2010-05-01

    The trade-offs among various output fidelities of asymmetric universal cloning machines are investigated. First we find out all the attainable optimal output fidelities for the 1 to 3 asymmetric universal cloning machine and it turns out that there are two kinds of extremal machines which have to cooperate in order to achieve some of the optimal output fidelities. Second we construct a family of extremal cloning machines that includes the universal symmetric cloning machine as well as an asymmetric 1 to 1+N cloning machine for qudits with two different output fidelities such that the optimal trade-off between the measurement disturbance and state estimation is attained in the limit of infinite N.

  20. PS2-47: Systematic Stakeholder Assessment to Determine Facilitators of and Barriers to SBIRT Implementation in Multiple Integrated Health Systems

    Science.gov (United States)

    Rahm, Alanna Kulchak; Boggs, Jennifer; Martin, Carmen; Beck, Arne; Pearson, Marilyn; Backer, Thomas; Ahmedani, Brian

    2014-01-01

    Background/Aims The US Preventive Services Task Force has identified alcohol and drug use screening as a high priority in primary care. SBIRT is a framework for population based screening and brief intervention with the goal of reducing substance use before it progresses to dependence. There has been limited uptake of SBIRT in any large health system, thus SAMHSA has supported a series of studies, led by Kaiser Permanente Colorado (KPCO), to determine strategies to promote large scale SBIRT implementation. Methods A qualitative examination of SBIRT feasibility and subsequent pilot testing within KPCO primary care guided a series of additional stakeholder interviews at Henry Ford Health System (HFHS) to assess feasibility of SBIRT implementation in additional organizations. Qualitative interviews were conducted with a broad spectrum of HFHS stakeholders using snowball sampling. A total of 28 HFHS stakeholders were interviewed. Interviews were conducted telephonically by trained interviewers from KPCO and lasted approximately 30 minutes. Interviews were digitally recorded and transcribed. After each interview, a detailed summary was generated and used to create the initial code book for analysis of interview transcripts. Results While current screening, brief intervention, and referral to treatment processes for substance use were noted in HFHS, they were inconsistent and no systematic policies exist. Many facilitators for SBIRT implementation within HFHS were noted including: population, recent implementation of depression screening in primary care, and EMR tools to facilitate documentation. Barriers to SBIRT implementation included time and competing demands for primary care providers, access to treatment, and follow-up processes for positive screens. All stakeholders were supportive of pilot implementation of SBIRT within HFHS with adequate attention to barriers and facilitators. Conclusions This project successfully demonstrated the value of gaining stakeholder

  1. A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.

    Science.gov (United States)

    Fowler, Daniel K; Stewart, Scott; Seredick, Steve; Eisen, Judith S; Stankunas, Kryn; Washbourne, Philip

    2016-01-01

    Recombination-based cloning is a quick and efficient way to generate expression vectors. Recent advancements have provided powerful recombinant DNA methods for molecular manipulations. Here, we describe a novel collection of three-fragment MultiSite Gateway cloning system-compatible vectors providing expanded molecular tools for vertebrate research. The components of this toolkit encompass a broad range of uses such as fluorescent imaging, dual gene expression, RNA interference, tandem affinity purification, chemically-inducible dimerization and lentiviral production. We demonstrate examples highlighting the utility of this toolkit for producing multi-component vertebrate expression vectors with diverse primary research applications. The vectors presented here are compatible with other Gateway toolkits and collections, facilitating the rapid generation of a broad range of innovative DNA constructs for biological research.

  2. A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.

    Directory of Open Access Journals (Sweden)

    Daniel K Fowler

    Full Text Available Recombination-based cloning is a quick and efficient way to generate expression vectors. Recent advancements have provided powerful recombinant DNA methods for molecular manipulations. Here, we describe a novel collection of three-fragment MultiSite Gateway cloning system-compatible vectors providing expanded molecular tools for vertebrate research. The components of this toolkit encompass a broad range of uses such as fluorescent imaging, dual gene expression, RNA interference, tandem affinity purification, chemically-inducible dimerization and lentiviral production. We demonstrate examples highlighting the utility of this toolkit for producing multi-component vertebrate expression vectors with diverse primary research applications. The vectors presented here are compatible with other Gateway toolkits and collections, facilitating the rapid generation of a broad range of innovative DNA constructs for biological research.

  3. Human cloning: can it be made safe?

    Science.gov (United States)

    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?

  4. Laser facilitates vaccination

    Directory of Open Access Journals (Sweden)

    Ji Wang

    2016-01-01

    Full Text Available Development of novel vaccine deliveries and vaccine adjuvants is of great importance to address the dilemma that the vaccine field faces: to improve vaccine efficacy without compromising safety. Harnessing the specific effects of laser on biological systems, a number of novel concepts have been proposed and proved in recent years to facilitate vaccination in a safer and more efficient way. The key advantage of using laser technology in vaccine delivery and adjuvantation is that all processes are initiated by physical effects with no foreign chemicals administered into the body. Here, we review the recent advances in using laser technology to facilitate vaccine delivery and augment vaccine efficacy as well as the underlying mechanisms.

  5. Limitations on Cloning in Classical Mechanics

    OpenAIRE

    Fenyes, Aaron

    2010-01-01

    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.

  6. Limitations on cloning in classical mechanics

    Science.gov (United States)

    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. Integration of an optical coherence tomography (OCT) system into a new environmental chamber to facilitate long term in vivo imaging of cardiovascular development in higher vertebrate embryos

    DEFF Research Database (Denmark)

    Thrane, Lars; Happel, Christoph M.; Thommes, Jan

    2010-01-01

    development. Here we demonstrate, to the best of our knowledge, the first realization of an optical coherence tomography (OCT) system integrated into a new environmental incubation chamber (EIC) to facilitate real-time in vivo imaging of cardiovascular development in chick embryos. The EIC provides stable...... conditions for embryonic development with respect to temperature, humidity, and oxygen levels. An OCT probe is integrated into the EIC and facilitates visualization of embryos at micrometer resolution, including the acquisition of M-mode, Doppler OCT, and Doppler M-mode data.......High-resolution 3-D in vivo imaging of embryonic development over long periods of time under constant physiological conditions (e.g. temperature, humidity) was a challenging task for researchers working on early cardiovascular development. Without appropriate maintenance of temperature, for example...

  8. Understanding barriers and facilitators to the use of Clinical Information Systems for intensive care units and Anesthesia Record Keeping: A rapid ethnography.

    Science.gov (United States)

    Saleem, Jason J; Plew, William R; Speir, Ross C; Herout, Jennifer; Wilck, Nancy R; Ryan, Dale Marie; Cullen, Theresa A; Scott, Jean M; Beene, Murielle S; Phillips, Toni

    2015-07-01

    This study evaluated the current use of commercial-off-the-shelf Clinical Information Systems (CIS) for intensive care units (ICUs) and Anesthesia Record Keeping (ARK) for operating rooms and post-anesthesia care recovery settings at three Veterans Affairs Medical Centers (VAMCs). Clinicians and administrative staff use these applications at bedside workstations, in operating rooms, at nursing stations, in physician's rooms, and in other various settings. The intention of a CIS or an ARK system is to facilitate creation of electronic records of data, assessments, and procedures from multiple medical devices. The US Department of Veterans Affairs (VA) Office of the Chief of Nursing Informatics sought to understand usage barriers and facilitators to optimize these systems in the future. Therefore, a human factors study was carried out to observe the CIS and ARK systems in use at three VAMCs in order to identify best practices and suggested improvements to currently implemented CIS and ARK systems. We conducted a rapid ethnographic study of clinical end-users interacting with the CIS and ARK systems in the critical care and anesthesia care areas in each of three geographically distributed VAMCs. Two observers recorded interactions and/or interview responses from 88 CIS and ARK end-users. We coded and sorted into logical categories field notes from 69 shadowed participants. The team transcribed and combined data from key informant interviews with 19 additional participants with the observation data. We then integrated findings across observations into meaningful patterns and abstracted the data into themes, which translated directly to barriers to effective adoption and optimization of the CIS and ARK systems. Effective optimization of the CIS and ARK systems was impeded by: (1) integration issues with other software systems; (2) poor usability; (3) software challenges; (4) hardware challenges; (5) training concerns; (6) unclear roles and lack of coordination among

  9. Barriers, facilitators and views about next steps to implementing supports for evidence-informed decision-making in health systems: a qualitative study.

    Science.gov (United States)

    Ellen, Moriah E; Léon, Grégory; Bouchard, Gisèle; Ouimet, Mathieu; Grimshaw, Jeremy M; Lavis, John N

    2014-12-05

    Mobilizing research evidence for daily decision-making is challenging for health system decision-makers. In a previous qualitative paper, we showed the current mix of supports that Canadian health-care organizations have in place and the ones that are perceived to be helpful to facilitate the use of research evidence in health system decision-making. Factors influencing the implementation of such supports remain poorly described in the literature. Identifying the barriers to and facilitators of different interventions is essential for implementation of effective, context-specific, supports for evidence-informed decision-making (EIDM) in health systems. The purpose of this study was to identify (a) barriers and facilitators to implementing supports for EIDM in Canadian health-care organizations, (b) views about emerging development of supports for EIDM, and (c) views about the priorities to bridge the gaps in the current mix of supports that these organizations have in place. This qualitative study was conducted in three types of health-care organizations (regional health authorities, hospitals, and primary care practices) in two Canadian provinces (Ontario and Quebec). Fifty-seven in-depth semi-structured telephone interviews were conducted with senior managers, library managers, and knowledge brokers from health-care organizations that have already undertaken strategic initiatives in knowledge translation. The interviews were taped, transcribed, and then analyzed thematically using NVivo 9 qualitative data analysis software. Limited resources (i.e., money or staff), time constraints, and negative attitudes (or resistance) toward change were the most frequently identified barriers to implementing supports for EIDM. Genuine interest from health system decision-makers, notably their willingness to invest money and resources and to create a knowledge translation culture over time in health-care organizations, was the most frequently identified facilitator to

  10. Facilitated saliva secretion and reduced oral inflammation by a novel artificial saliva system in the treatment of salivary hypofunction

    Directory of Open Access Journals (Sweden)

    Kang M

    2017-01-01

    Full Text Available Minkyung Kang,1 Hyounggeun Park,1 Joon-Ho Jun,1 Miwon Son,1 Myung Joo Kang2 1Pharmaceutical Product Research Laboratories, Dong-A ST Research Institute, Gyeonggi, 2Division of Pharmaceutical Sciences, College of Pharmacy, Dankook University, Cheonan, Chungnam, Korea Abstract: Saliva substitutes and/or lubricants are commonly employed to lessen dry mouth symptoms by stimulating and/or substituting for the secretion of saliva. In this study, a novel artificial saliva containing inorganic salts, including sodium chloride and potassium chloride, and bactericidal agents, including potassium thiocyanate and lactoperoxidase, was formulated in the form of a solution (DM-sol or gel (DM-gel. Those in vivo therapeutic efficacies were assessed in terms of saliva secretion and anti-inflammatory activity in rats and mice, respectively. Salivary secretion was promoted by mucosal application of DM-formulations in normal rats. In particular, DM-gel resulted in 2.5- and 1.9-fold greater salivary flow rates compared to normal saline and DM-sol, respectively. In an in vivo efficacy evaluation in diabetic mice with salivary hypofunction, repeated application of DM-formulations alleviated histopathological changes in the buccal mucosa in terms of atrophy and thinning of the epithelium, compared to vehicle, after 4 weeks. Moreover, the DM-sol and DM-gel were comparably effective for relieving periodontal gingivitis, reducing infiltration of inflammatory cells, and normalizing the neutrophil level in the gingival gingiva, after 4 weeks. Therefore, the novel artificial saliva is expected to facilitate salivary secretion and restore physiological conditions in the mouth of patients with salivary hypofunction. Keywords: saliva substitute, carbopol gel, hypothiocyanite–hydrogen peroxide mixture, antimicrobial activity, diabetic rats

  11. Human cloning and 'posthuman' society.

    Science.gov (United States)

    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.

  12. Human Cloning: Let's Discuss It.

    Science.gov (United States)

    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)

  13. A Clone of Your Own.

    Science.gov (United States)

    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)

  14. A strategy for clone selection under different production conditions.

    Science.gov (United States)

    Legmann, Rachel; Benoit, Brian; Fedechko, Ronald W; Deppeler, Cynthia L; Srinivasan, Sriram; Robins, Russell H; McCormick, Ellen L; Ferrick, David A; Rodgers, Seth T; Russo, A Peter

    2011-01-01

    Top performing clones have failed at the manufacturing scale while the true best performer may have been rejected early in the screening process. Therefore, the ability to screen multiple clones in complex fed-batch processes using multiple process variations can be used to assess robustness and to identify critical factors. This dynamic ranking of clones' strategy requires the execution of many parallel experiments than traditional approaches. Therefore, this approach is best suited for micro-bioreactor models which can perform hundreds of experiments quickly and efficiently. In this study, a fully monitored and controlled small scale platform was used to screen eight CHO clones producing a recombinant monoclonal antibody across several process variations, including different feeding strategies, temperature shifts and pH control profiles. The first screen utilized 240 micro-bioreactors were run for two weeks for this assessment of the scale-down model as a high-throughput tool for clone evaluation. The richness of the outcome data enable to clearly identify the best and worst clone as well as process in term of maximum monoclonal antibody titer. The follow-up comparison study utilized 180 micro-bioreactors in a full factorial design and a subset of 12 clone/process combinations was selected to be run parallel in duplicate shake flasks. Good correlation between the micro-bioreactor predictions and those made in shake flasks with a Pearson correlation value of 0.94. The results also demonstrate that this micro-scale system can perform clone screening and process optimization for gaining significant titer improvements simultaneously. This dynamic ranking strategy can support better choices of production clones. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

  15. Artificial cloning of domestic animals.

    Science.gov (United States)

    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.

  16. Islamic perspectives on human cloning.

    Science.gov (United States)

    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.

  17. Cloning goes to the movies.

    Science.gov (United States)

    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.

  18. QuaDoSta - a freely configurable system which facilitates multi-centric data collection for healthcare and medical research

    Directory of Open Access Journals (Sweden)

    Albrecht, Ulrike

    2007-07-01

    Full Text Available This article describes QuaDoSta (quality assurance, documentation and statistics, a flexible documentation system as well as a data collection and networking platform for medical facilities. The user can freely define the required documentation masks which are easily expandable and can be adapted to individual requirements without the need for additional programming. To avoid duplication, data transfer interfaces can be configured flexibly to external sources such as patient management systems used in surgeries or hospital information systems. The projects EvaMed (Evaluation Anthroposophical Medicine and the Network Oncology are two scientific research projects which have been successfully established as nationally active networks on the basis of QuaDoSta. The EvaMed-Network serves as a modern pharmacovigilance project for the documentation of adverse drug events. All prescription data are electronically recorded to assess the relative risk of drugs. The Network Oncology was set up as a documentation system in four hospitals and seven specialist oncology practices where a complete record of all oncological therapies is being carried out to uniform standards on the basis of the ‘basic documentation for tumour patients’ (BDT developed by the German Cancer Society. The QuaDoSta solution system made it possible to cater for the specific requirements of the presented projects. The following features of the system proved to be highly advantageous: flexible setup of catalogues and user friendly customisation and extensions, complete dissociation of system setup and documentation content, multi-centre networkability, and configurable data transfer interfaces.

  19. Expression of MexAB-OprM efflux pump system and susceptibility to antibiotics of different Pseudomonas aeruginosa clones isolated from patients hospitalized in two intensive care units at University Hospital in Bialystok (northeastern Poland) between January 2002 and December 2009.

    Science.gov (United States)

    Sacha, Paweł; Wieczorek, Piotr; Ojdana, Dominika; Hauschild, Tomasz; Milewski, Robert; Czaban, Sławomir; Poniatowski, Bogusław; Tryniszewska, Elżbieta

    2014-10-01

    We investigated the genetic similarities and expression of the MexAB-OprM efflux pump system in different clones of multiresistant Pseudomonas aeruginosa strains collected from 2002 to 2009 at two intensive care units (ICU). Regulatory and structural genes mexB, mexR, and mexA were found in 99%, 98%, and 94% of tested strains, respectively. The presence of class 1 integron was found in 90% of the strains, while class 2 integron in only one strain (Psa506). Class 3 integron was not found in any of the tested strains. Among the eleven clones identified, only two clones, I and D, exhibited higher levels of mexB gene expression than the other clones. Clone I had the highest expression (FC = 10.36, p resistance to all tested antibiotics among the various clones. The high level of antimicrobial resistance may have been due to the coexistence of different resistance mechanisms among the studied P. aeruginosa strains. However, this does not exclude the contribution of the MexAB-OprM pump, particularly in resistance to meropenem and ciprofloxacin. © 2014 APMIS. Published by John Wiley & Sons Ltd.

  20. Structured Review of Code Clone Literature

    NARCIS (Netherlands)

    Hordijk, W.T.B.; Ponisio, Laura; Wieringa, Roelf J.

    2008-01-01

    This report presents the results of a structured review of code clone literature. The aim of the review is to assemble a conceptual model of clone-related concepts which helps us to reason about clones. This conceptual model unifies clone concepts from a wide range of literature, so that findings

  1. Quantum cloning machines for equatorial qubits

    International Nuclear Information System (INIS)

    Fan Heng; Matsumoto, Keiji; Wang Xiangbin; Wadati, Miki

    2002-01-01

    Quantum cloning machines for equatorial qubits are studied. For the case of a one to two phase-covariant quantum cloning machine, we present the networks consisting of quantum gates to realize the quantum cloning transformations. The copied equatorial qubits are shown to be separable by using Peres-Horodecki criterion. The optimal one to M phase-covariant quantum cloning transformations are given

  2. An Improved Homologous Recombination Method for Rapid Cloning of the Antibody Heavy Chain Gene and Its Comparison with the Homologous Recombination and Traditional Cloning Methods

    Directory of Open Access Journals (Sweden)

    Masoumeh Hajirezaei

    2015-10-01

    Full Text Available Background: The homologous recombination (HR is one of the conventional cloning methods for the production of recombinant DNA. It is a quick method for in vivo DNA cloning without using the high cost restriction enzymes. A few modifications in the cloning procedure can increase the efficiency of this method.Materials and Methods: In this study, effect of heating on the rate of the IgG1 heavy chain gene cloning was investigated in the HR method and then it was compared with HR method without heating and traditional cloning method. For doing this comparison, three cloning methods including HR, HR with the heat treatment, and traditional cloning were used to clone the human IgG1 heavy chain into the pcDNA3.1(+ plasmid.Results: The results showed that adding heat in the HR method converts insert and vector from the double strand DNA to the single strand DNA. This allows them to copulate with each other better and faster than the two other methods. The heat addition also helps the cell enzyme system for a faster and easier recombination and moreover it increases the cloning efficiency of the HR method in case of in vitro processing.Conclusion: The results showed that giving heat in the HR method increases cloning rate 7.5% and this increase reaches 10% in comparison with the traditional method. 

  3. Methylotroph cloning vehicle

    Science.gov (United States)

    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.

  4. Cloning and Distribution of a Putative Octopamine/Tyramine Receptor in the Central Nervous System of the Freshwater Prawn Macrobrachium rosenbergii

    Science.gov (United States)

    Reyes-Colón, Dalynés; Vázquez-Acevedo, Nietzell; Rivera, Nilsa M.; Jezzini, Sami H.; Rosenthal, Joshua; Ruiz-Rodríguez, Eduardo A.; Baro, Deborah J.; Kohn, Andrea B.; Moroz, Leonid; Sosa, María A.

    2010-01-01

    There is ample evidence linking octopamine (OA) and tyramine (TA) to several neurophysiological functions in arthropods. In our laboratory we use the freshwater prawn Macrobrachium rosenbergii to study the neural basis of aggressive behavior. As a first step towards understanding the possible role of these amines and their receptors in the modulation of interactive behaviors, we have cloned a putative octopamine/tyramine receptor. The predicted sequence of the cloned OA/TAMac receptor consists of 1,579 base pairs (bp), with an open reading frame of 1,350 bp that encodes a 450 amino acid protein. This putative protein displays sequence identities of 70% to an Aedes aegypti mosquito TA receptor, followed by 60% to a Stegomyia aegypti mosquito OA receptor, 59% and 58% to the migratory locust TA-1 and -2 receptors respectively, and 57% with the silkworm OA receptor. We also mapped the OA/TAMac receptor distribution by in-situ hybridization to the receptor’s mRNA, and by immunohistochemistry to its protein. We observed stained cell bodies for the receptor’s mRNA, mainly in the midline region of the thoracic and in the abdominal ganglia, as well as diffuse staining in the brain ganglia. For the receptor’s protein, we observed extensive punctate staining within the neuropil and on the membrane of specific groups of neurons in all ganglia throughout the CNS, including the brain, the midline region and neuropiles of the thoracic ganglia, and ventral part and neuropiles of the abdominal ganglia. The same pattern of stained cells was observed on the thoracic and abdominal ganglia in both in-situ hybridization and immunohistochemistry experiments. Diffuse staining observed with in-situ hybridization also coincides with punctate staining observed in brain, SEG, thoracic, and abdominal ganglia in immunohistochemical preparations. This work provides the first step towards characterizing the neural networks that mediate octopaminergic signaling in prawn. PMID:20558147

  5. JUICE: a data management system that facilitates the analysis of large volumes of information in an EST project workflow

    Directory of Open Access Journals (Sweden)

    Martinez Veronica

    2006-11-01

    Full Text Available Abstract Background Expressed sequence tag (EST analyses provide a rapid and economical means to identify candidate genes that may be involved in a particular biological process. These ESTs are useful in many Functional Genomics studies. However, the large quantity and complexity of the data generated during an EST sequencing project can make the analysis of this information a daunting task. Results In an attempt to make this task friendlier, we have developed JUICE, an open source data management system (Apache + PHP + MySQL on Linux, which enables the user to easily upload, organize, visualize and search the different types of data generated in an EST project pipeline. In contrast to other systems, the JUICE data management system allows a branched pipeline to be established, modified and expanded, during the course of an EST project. The web interfaces and tools in JUICE enable the users to visualize the information in a graphical, user-friendly manner. The user may browse or search for sequences and/or sequence information within all the branches of the pipeline. The user can search using terms associated with the sequence name, annotation or other characteristics stored in JUICE and associated with sequences or sequence groups. Groups of sequences can be created by the user, stored in a clipboard and/or downloaded for further analyses. Different user profiles restrict the access of each user depending upon their role in the project. The user may have access exclusively to visualize sequence information, access to annotate sequences and sequence information, or administrative access. Conclusion JUICE is an open source data management system that has been developed to aid users in organizing and analyzing the large amount of data generated in an EST Project workflow. JUICE has been used in one of the first functional genomics projects in Chile, entitled "Functional Genomics in nectarines: Platform to potentiate the competitiveness of Chile in

  6. ClimateQUAL® and Thinklets: Using ClimateQUAL® with Group Support Systems to Facilitate Discussion and Set Priorities for Organizational Change at Criss Library

    Directory of Open Access Journals (Sweden)

    Nora Hillyer

    2013-06-01

    Full Text Available Objective – This article discusses a series of actions taken by the Criss Library at the University of Nebraska at Omaha to implement organizational change, using the ClimateQUAL® survey and facilitated discussions with ThinkTank™ group decision software. The library had experienced significant changes over a five-year period, with a renovation of the facility and three reorganizations resulting in a 50% staff turnover. Recognizing the strain that years of construction and personnel changes had placed on the organization, there was a desire to uncover the mood of the employees and reveal the issues behind low morale, uneasiness, and fear.Methods – In November 2009, the library conducted a ClimateQUAL® survey to develop a baseline to assess the effectiveness of any changes. After the results were distributed to library faculty and staff, a series of two-hour facilitated discussions was held to gather opinions and ideas for solutions using thinkLets, a pattern language for reasoning toward a goal. The group support system ThinkTank™ software was loaded onto computers, and employees were able to add their ideas anonymously during the sessions. Finally, 12 employees (29% completed a four-question survey on their perceptions of the facilitated discussions.Results – The facilitated discussions returned 76 sub-themes in 12 categories: staffing and scheduling issues, staff unity/teamwork, communication, goodwill/morale, accountability, decision-making, policy issues, skills and training, leadership, ergonomics/physical work environment, respect, and bullying. An advisory team culled the 76 sub-themes into 40 improvement strategies. Five were implemented immediately, and the remaining 35 were scheduled to be presented to the faculty and staff via an online survey. Participants’ perceptions of the facilitated discussions were mixed. Eighty-three percent of respondents reported that they did not feel safe speaking out about issues, most

  7. Facilitating School Change Using the Change Communication Model: The Adoption and Implementation of a New Student Information System

    Science.gov (United States)

    Rutschke, Linda Louise

    2010-01-01

    This study examines how the Change Communication Model can be used to implement a new student information system (SIS) in a school district. The impetus of the study came from government mandates requiring district accountability through data-driven decision making. Data-driven decision making is only possible when student data are collected,…

  8. Development of a Matlab/Simulink tool to facilitate system analysis and simulation via the adjoint and covariance methods

    NARCIS (Netherlands)

    Bucco, D.; Weiss, M.

    2007-01-01

    The COVariance and ADjoint Analysis Tool (COVAD) is a specially designed software tool, written for the Matlab/Simulink environment, which allows the user the capability to carry out system analysis and simulation using the adjoint, covariance or Monte Carlo methods. This paper describes phase one

  9. Local cloning of two product states

    International Nuclear Information System (INIS)

    Ji Zhengfeng; Feng Yuan; Ying Mingsheng

    2005-01-01

    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

  10. CRISPR-Cas9-Edited Site Sequencing (CRES-Seq): An Efficient and High-Throughput Method for the Selection of CRISPR-Cas9-Edited Clones.

    Science.gov (United States)

    Veeranagouda, Yaligara; Debono-Lagneaux, Delphine; Fournet, Hamida; Thill, Gilbert; Didier, Michel

    2018-01-16

    The emergence of clustered regularly interspaced short palindromic repeats-Cas9 (CRISPR-Cas9) gene editing systems has enabled the creation of specific mutants at low cost, in a short time and with high efficiency, in eukaryotic cells. Since a CRISPR-Cas9 system typically creates an array of mutations in targeted sites, a successful gene editing project requires careful selection of edited clones. This process can be very challenging, especially when working with multiallelic genes and/or polyploid cells (such as cancer and plants cells). Here we described a next-generation sequencing method called CRISPR-Cas9 Edited Site Sequencing (CRES-Seq) for the efficient and high-throughput screening of CRISPR-Cas9-edited clones. CRES-Seq facilitates the precise genotyping up to 96 CRISPR-Cas9-edited sites (CRES) in a single MiniSeq (Illumina) run with an approximate sequencing cost of $6/clone. CRES-Seq is particularly useful when multiple genes are simultaneously targeted by CRISPR-Cas9, and also for screening of clones generated from multiallelic genes/polyploid cells. © 2018 by John Wiley & Sons, Inc. Copyright © 2018 John Wiley & Sons, Inc.

  11. Development of Intelligent Computer-Assisted Instruction Systems to Facilitate Reading Skills of Learning-Disabled Children

    Science.gov (United States)

    1993-12-01

    people or mediating conflicts 17 between peers or family members. They tend to socialize and seem to know how people feel. Intrapersonal children...accessible to more schools and families , it will become easier to address more of the learning styles due to the expanding system capabilities while keeping...Annual Coastal Conference of the Orion Dyslexia Society. 23 Jan 1993. ARMS2 93 Armsstrong, Thomas. 7 Kim/v (Y’ Smart: idenijfting and IDevel pingq Your

  12. Implementing a web-based home monitoring system within an academic health care network: barriers and facilitators to innovation diffusion.

    Science.gov (United States)

    Pelletier, Alexandra C; Jethwani, Kamal; Bello, Heather; Kvedar, Joseph; Grant, Richard W

    2011-01-01

    The practice of outpatient type 2 diabetes management is gradually moving from the traditional visit-based, fee-for-service model to a new, health information communication technology (ICT)-supported model that can enable non-visit-based diabetes care. To date, adoption of innovative health ICT tools for diabetes management has been slowed by numerous barriers, such as capital investment costs, lack of reliable reimbursement mechanisms, design defects that have made some systems time-consuming and inefficient to use, and the need to integrate new ICT tools into a system not primarily designed for their use. Effective implementation of innovative diabetes health ICT interventions must address local practice heterogeneity and the interaction of this heterogeneity with clinical care delivery. The Center for Connected Health at Partners Healthcare has implemented a new ICT intervention, Diabetes Connect (DC), a Web-based glucose home monitoring and clinical messaging system. Using the framework of the diffusion of innovation theory, we review the implementation and examine lessons learned as we continue to deploy DC across the health care network. © 2010 Diabetes Technology Society.

  13. A synthetic three-dimensional niche system facilitates generation of functional hematopoietic cells from human-induced pluripotent stem cells

    Directory of Open Access Journals (Sweden)

    Yulin Xu

    2016-09-01

    Full Text Available Abstract Background The efficient generation of hematopoietic stem cells (HSCs from human-induced pluripotent stem cells (iPSCs holds great promise in personalized transplantation therapies. However, the derivation of functional and transplantable HSCs from iPSCs has had very limited success thus far. Methods We developed a synthetic 3D hematopoietic niche system comprising nanofibers seeded with bone marrow (BM-derived stromal cells and growth factors to induce functional hematopoietic cells from human iPSCs in vitro. Results Approximately 70 % of human CD34+ hematopoietic cells accompanied with CD43+ progenitor cells could be derived from this 3D induction system. Colony-forming-unit (CFU assay showed that iPSC-derived CD34+ cells formed all types of hematopoietic colonies including CFU-GEMM. TAL-1 and MIXL1, critical transcription factors associated with hematopoietic development, were expressed during the differentiation process. Furthermore, iPSC-derived hematopoietic cells gave rise to both lymphoid and myeloid lineages in the recipient NOD/SCID mice after transplantation. Conclusions Our study underscores the importance of a synthetic 3D niche system for the derivation of transplantable hematopoietic cells from human iPSCs in vitro thereby establishing a foundation towards utilization of human iPSC-derived HSCs for transplantation therapies in the clinic.

  14. The intervening removable affinity tag (iRAT) production system facilitates Fv antibody fragment‐mediated crystallography

    Science.gov (United States)

    Nomura, Yayoi; Sato, Yumi; Suno, Ryoji; Horita, Shoichiro

    2016-01-01

    Abstract Fv antibody fragments have been used as co‐crystallization partners in structural biology, particularly in membrane protein crystallography. However, there are inherent technical issues associated with the large‐scale production of soluble, functional Fv fragments through conventional methods in various expression systems. To circumvent these problems, we developed a new method, in which a single synthetic polyprotein consisting of a variable light (VL) domain, an intervening removable affinity tag (iRAT), and a variable heavy (VH) domain is expressed by a Gram‐positive bacterial secretion system. This method ensures stoichiometric expression of VL and VH from the monocistronic construct followed by proper folding and assembly of the two variable domains. The iRAT segment can be removed by a site‐specific protease during the purification process to yield tag‐free Fv fragments suitable for crystallization trials. In vitro refolding step is not required to obtain correctly folded Fv fragments. As a proof of concept, we tested the iRAT‐based production of multiple Fv fragments, including a crystallization chaperone for a mammalian membrane protein as well as FDA‐approved therapeutic antibodies. The resulting Fv fragments were functionally active and crystallized in complex with the target proteins. The iRAT system is a reliable, rapid and broadly applicable means of producing milligram quantities of Fv fragments for structural and biochemical studies. PMID:27595817

  15. Human Endothelial Cells: Use of Heparin in Cloning and Long-Term Serial Cultivation

    Science.gov (United States)

    Thornton, Susan C.; Mueller, Stephen N.; Levine, Elliot M.

    1983-11-01

    Endothelial cells from human blood vessels were cultured in vitro, with doubling times of 17 to 21 hours for 42 to 79 population doublings. Cloned human endothelial cell strains were established for the first time and had similar proliferative capacities. This vigorous cell growth was achieved by addition of heparin to culture medium containing reduced concentrations of endothelial cell growth factor. The routine cloning and long-term culture of human endothelial cells will facilitate studying the human endothelium in vitro.

  16. The characterization of an economic and portable LED-based photoacoustic imaging system to facilitate molecular imaging

    Directory of Open Access Journals (Sweden)

    Ali Hariri

    2018-03-01

    Full Text Available Photoacoustic imaging (PAI is a non-invasive, high-resolution hybrid imaging modality that combines optical excitation and ultrasound detection. PAI can image endogenous chromophores (melanin, hemoglobin, etc. and exogenous contrast agents in different medical applications. However, most current equipment uses sophisticated and complicated OPO lasers with tuning and stability features inconsistent with broad clinical deployment. As the number of applications of PAI in medicine increases, there is an urgent need to make the imaging equipment more compact, portable, and affordable. Here, portable light emitting diode – based photoacoustic imaging (PLED-PAI was introduced and characterized in terms of system specifications, light source characterizations, photoacoustic spatial/temporal resolution, and penetration. The system uses two LED arrays attached to the sides of a conventional ultrasound transducer. The LED pulse repetition rate is tunable between 1 K Hz, 2 K Hz, 3 K Hz, and 4 K Hz. The axial resolution was 0.268 mm, and the lateral resolution was between 0.55 and 0.59 mm. The system could detect optical absorber (pencil lead at a depth of 3.2 cm and the detection limits of indocyanine green (ICG and methylene blue (MB were 9 μM and 0.78 mM. In vivo imaging of labeled human mesenchymal stem cells was achieved to confirm compatibility with small animal imaging. The characterization we report here may have value to other groups evaluating commercially available photoacoustic imaging equipment. Keywords: Portable photoacoustic imaging, LED, Optoacoustic imaging, Molecular imaging

  17. The arginine deiminase system facilitates environmental adaptability of Streptococcus equi ssp. zooepidemicus through pH adjustment.

    Science.gov (United States)

    Xu, Bin; Yang, Xinyi; Zhang, Ping; Ma, Zhe; Lin, Huixing; Fan, Hongjie

    2016-06-01

    The arginine deiminase system (ADS) is a secondary metabolic system found in many different bacterial pathogens and it is often associated with virulence. Here, a systematic study of ADS functions in Streptococcus equi subsp. zooepidemicus (SEZ) was performed. Transcriptional levels of ADS operon genes were observed to be significantly increased when SEZ was grown under acidic conditions. We constructed arcA and arcD deletion mutants (SEZ ΔarcA and SEZ ΔarcD, respectively) and found that SEZ ΔarcA was unable to metabolize arginine and synthesize ammonia; however, arcD deletion resulted in an initial decrease in arginine consumption and ammonia production, followed by recovery to the levels of wild-type SEZ after 24 h of cultivation. Cell extracts of SEZ ΔarcA showed no arginine deiminase (AD) activity, whereas no difference in AD activity between SEZ ΔarcD and wild-type SEZ was observed. SEZ survival tests demonstrated a significant decrease in survival for SEZ ΔarcA, when compared with wild-type SEZ, under acidic conditions and in epithelial cells. These findings indicate that ADS in SEZ contributes to environmental adaptability via ammonia synthesis to reduce pH stress. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  18. TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.

    Directory of Open Access Journals (Sweden)

    Athanasios Niarchos

    Full Text Available During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, and T and C overhangs of the novel vector pET-BccI, created after digestion with the restriction endonuclease BccI. The novel protein-expression vector pET-BccI also facilitates the screening of transformed colonies for recombinant transformants. Evaluation experiments of the proposed TA-GC cloning method showed that 81% of the transformed colonies contained recombinant pET-BccI plasmids, and 98% of the recombinant colonies expressed the desired protein. This demonstrates that TA-GC cloning could be a valuable method for cloning protein-coding genes in expression vectors.

  19. Trade patterns facilitating highly pathogenic avian influenza virus dissemination in the free-grazing layer duck system in Vietnam.

    Science.gov (United States)

    Meyer, A; Dinh, T X; Han, T A; Do, D V; Nhu, T V; Pham, L T; Nguyen, T T T; Newman, S; Häsler, B; Pfeiffer, D U; Vergne, T

    2018-04-01

    Highly pathogenic avian influenza (HPAI) viruses continue to threaten smallholder poultry producers in several South-east Asian countries, including Vietnam. In particular, the free-grazing duck system has been repeatedly highlighted as a major risk factor for HPAI outbreaks. Free-grazing ducks, which scavenge on rice paddies after the harvest, account for a large proportion of the duck population in Vietnam and the wider South-east Asian region. However, the structure and dynamics of the free-grazing duck production from farm to consumption has not been described for Vietnam. In this study, we used a value chain approach to provide a complete picture of the actors involved in the production and marketing of free-grazing duck eggs and spent layer ducks, as well as to investigate the governance structure of this food system. Group interviews and key informant interviews were conducted in two provinces located in the Mekong River Delta (MRD) and the Red River Delta (RRD). The results presented here highlight similarities and differences in farming and trade practices between the two provinces. The trade of spent layer ducks involved large volumes of live ducks being sent to China and Cambodia for consumption, generating a substantial risk of transboundary spread of pathogens, including HPAI viruses. We describe the major role of "duck yards", which act as hubs in the northbound trade of spent layer ducks. These yards should be considered as essential links in the value chain of spent layer ducks when considering HPAI surveillance and control. The veterinary authorities are only marginally involved in the value chain activities, and their influence could be strengthened by increasing surveillance activities for instance in duck yards. Last, we discuss the dynamics of the duck value chain and further implications for future HPAI management policies. © 2017 The Authors. Transboundary and Emerging Diseases Published by Blackwell Verlag GmbH.

  20. A miRNA-responsive cell-free translation system facilitates isolation of hepatitis C virus miRNP complexes.

    Science.gov (United States)

    Bradrick, Shelton S; Nagyal, Simardeep; Novatt, Hilary

    2013-08-01

    Micro(mi)RNAs are 21- to 23-nt RNAs that regulate multiple biological processes. In association with Argonaute (Ago) proteins and other factors that form the RNA-induced silencing complex (RISC), miRNAs typically bind mRNA 3' untranslated regions (UTRs) and repress protein production through antagonizing translation and transcript stability. For a given mRNA-miRNA interaction, cis-acting RNA elements and trans-acting RNA-binding proteins (RBPs) may influence mRNA fate. This is particularly true of the hepatitis C virus (HCV) genome which interacts with miR-122, an abundant liver miRNA. miR-122 binding to HCV RNA considerably stimulates virus replication in cultured cells and primates, but the mechanism(s) and associated host factors required for enhancement of HCV replication have not been fully elucidated. We recapitulated miR-122-HCV RNA interactions in a cell-free translation system derived from cells that express miR-122. Specifically, lysates produced from HEK-293 cells that inducibly transcribe and process pri-miR-122 were characterized alongside those from isogenic cells lacking miR-122 expression. We observed a stimulatory effect of miR-122 on HCV reporter mRNAs in a manner that depended on expression of miR-122 and intact target sites within the HCV 5' UTR. We took advantage of this system to affinity-purify miR-122-HCV RNP complexes. Similar to functional assays, we found that association of immobilized HCV internal ribosome entry site (IRES) RNA with endogenous Ago2 requires both miR-122 expression and intact miR-122 target sites in cis. This combined approach may be generalizable to affinity purification of miRNP complexes for selected target mRNAs, allowing identification of miRNP components and RBPs that may contribute to regulation.

  1. Identification of an Acinetobacter baumannii zinc acquisition system that facilitates resistance to calprotectin-mediated zinc sequestration.

    Directory of Open Access Journals (Sweden)

    M Indriati Hood

    Full Text Available Acinetobacter baumannii is an important nosocomial pathogen that accounts for up to 20 percent of infections in intensive care units worldwide. Furthermore, A. baumannii strains have emerged that are resistant to all available antimicrobials. These facts highlight the dire need for new therapeutic strategies to combat this growing public health threat. Given the critical role for transition metals at the pathogen-host interface, interrogating the role for these metals in A. baumannii physiology and pathogenesis could elucidate novel therapeutic strategies. Toward this end, the role for calprotectin- (CP-mediated chelation of manganese (Mn and zinc (Zn in defense against A. baumannii was investigated. These experiments revealed that CP inhibits A. baumannii growth in vitro through chelation of Mn and Zn. Consistent with these in vitro data, Imaging Mass Spectrometry revealed that CP accompanies neutrophil recruitment to the lung and accumulates at foci of infection in a murine model of A. baumannii pneumonia. CP contributes to host survival and control of bacterial replication in the lung and limits dissemination to secondary sites. Using CP as a probe identified an A. baumannii Zn acquisition system that contributes to Zn uptake, enabling this organism to resist CP-mediated metal chelation, which enhances pathogenesis. Moreover, evidence is provided that Zn uptake across the outer membrane is an energy-dependent process in A. baumannii. Finally, it is shown that Zn limitation reverses carbapenem resistance in multidrug resistant A. baumannii underscoring the clinical relevance of these findings. Taken together, these data establish Zn acquisition systems as viable therapeutic targets to combat multidrug resistant A. baumannii infections.

  2. Facilitating Deployment of Community Solar PV systems on Rooftops and Vacant Land in Northeast IL - Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Stone, Deborah [Cook County, Chicago, IL (United States); Oakleaf, Laura [Cook County, Chicago, IL (United States)

    2017-12-31

    The Cook County Community Solar project set out to unlock the potential of community solar in the Chicago region with lessons that could be applied nationally. One of the first steps was to prove out the potential market. This was done through an opportunity assessment which showed there is over 9,000 megawatts worth of site capacity available for community solar projects in Cook County – nearly enough to offset all of Cook County’s residential electricity use. The assessment also showed that almost 75% of Cook County households are not able to invest directly in solar photovoltaic systems due to a variety of issues from physical barriers such as shading, or condition of the roof, to financial barriers such as lack of roof ownership, or the up-front costs of installation. Because of these barriers, community solar is an essential part of making the benefits of renewable energy available to all of the residents of Cook County. In addition to the opportunity assessment the project team also worked with the over 200 individuals who participated in the stakeholder advisory group to develop a number of other products including: 1) an Economic & Policy Barriers Resolutions and Work Plan document which laid out best practices to address the policy barriers that existed at the time (May of 2016) 2) Value Proposition Report I and Report II which summarize the value of community solar to potential developers and subscribers, 3) The Community Solar Business Case Tool, which provides a flexible financial model that projects the costs and befits to the system developer and subscriber for a project, 4) Bill Crediting Analysis and the 5) Final Report. The Final Report contains 15 case studies which prove that community solar projects are economically feasible in Cook County with a variety of sites, solar designs, ownership and subscriber models.

  3. [Mystery and problems of cloning].

    Science.gov (United States)

    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.

  4. [Cloning and law in Hungary].

    Science.gov (United States)

    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.

  5. Quantum cloning without external control

    International Nuclear Information System (INIS)

    Chiara, G. de; Fazio, R.; Macchiavello, C.; Montangero, S.; Palma, G.M.

    2005-01-01

    Full text: In this work we present an approach to quantum cloning with unmodulated spin networks. The cloner is realized by a proper design of the network and a choice of the coupling between the qubits. We show that in the case of phase covariant cloner the XY coupling gives the best results. In the 1 → 2 cloning we find that the value for the fidelity of the optimal cloner is achieved, and values comparable to the optimal ones in the general N → M case can be attained. If a suitable set of network symmetries are satisfied, the output fidelity of the clones does not depend on the specific choice of the graph. We show that spin network cloning is robust against the presence of static imperfections. Moreover, in the presence of noise, it outperforms the conventional approach. In this case the fidelity exceeds the corresponding value obtained by quantum gates even for a very small amount of noise. Furthermore we show how to use this method to clone qutrits and qudits. By means of the Heisenberg coupling it is also possible to implement the universal cloner although in this case the fidelity is 10 % off that of the optimal cloner. (author)

  6. Determining Complementary Properties with Quantum Clones

    Science.gov (United States)

    Thekkadath, G. S.; Saaltink, R. Y.; Giner, L.; Lundeen, J. S.

    2017-08-01

    In a classical world, simultaneous measurements of complementary properties (e.g., position and momentum) give a system's state. In quantum mechanics, measurement-induced disturbance is largest for complementary properties and, hence, limits the precision with which such properties can be determined simultaneously. It is tempting to try to sidestep this disturbance by copying the system and measuring each complementary property on a separate copy. However, perfect copying is physically impossible in quantum mechanics. Here, we investigate using the closest quantum analog to this copying strategy, optimal cloning. The coherent portion of the generated clones' state corresponds to "twins" of the input system. Like perfect copies, both twins faithfully reproduce the properties of the input system. Unlike perfect copies, the twins are entangled. As such, a measurement on both twins is equivalent to a simultaneous measurement on the input system. For complementary observables, this joint measurement gives the system's state, just as in the classical case. We demonstrate this experimentally using polarized single photons.

  7. The topsy-turvy cloning law.

    Science.gov (United States)

    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.

  8. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2010-01-01

    Code clones - that is, duplicate fragments of code - have been studied for a long time. There is strong evidence that code clones are a major source of software faults. Anecdotal evidence suggests that this phenomenon is not restricted to code, but occurs in models in a very similar way. So...... it is likely that model clones are as detrimental to model quality as they are to code quality. However, programming language code and visual models also have significant differences so that notions and algorithms developed in the code clone arena cannot be transferred directly to model clones. In this article......, we discuss how model clones arise by analyzing several practical scenarios. We propose a formal definition of models and clones, that allows us to specify a generic clone detection algorithm. Through a thorough analysis of the detail structure of sample UML domain models, recommendations for clone...

  9. Core Structure Elements Architectures to Facilitate Construction and Secure Interconnection of Mobile Services Frameworks and Advanced IAM Systems

    Science.gov (United States)

    Karantjias, Athanasios; Polemi, Nineta

    The impressing penetration rates of electronic and mobile networks provide the unique opportunity to organizations to provide advanced e/m-services, accelerating their entrance in the digital society, and strengthening their fundamental structure. Service Oriented Architectures (SOAs) is an acknowledged promising technology to overcome the complexity inherent to the communication among multiple e-business actors across organizational domains. Nevertheless, the need for more privacy-aware transactions raises specific challenges that SOAs need to address, including the problems of managing identities and ensuring privacy in the e/m-environment. This article presents a targeted, user-centric scalable and federated Identity Management System (IAM), calledSecIdAM, and a mobile framework for building privacy-aware, interoperable, and secure mobile applications with respect to the way that the trust relationship among the involved entities, users and SOAs, is established. Finally, it analyzes a user-transparent m-process for obtaining an authentication and authorization token, issued from the SecIdAM as integrated in the IST European programme SWEB for the public sector.

  10. Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system.

    Science.gov (United States)

    Chen, Haide; Jiang, Mengmeng; Xiao, Lei; Huang, He

    2018-03-15

    Human pluripotent stem cells (hPSCs)/OP9 coculture system is a widely used hematopoietic differentiation approach. The limited understanding of this process leads to its low efficiency. Thus, we used single-cell qPCR to reveal the gene expression profiles of individual CD34+ cells from different stages of differentiation. According to the dynamic gene expression of hematopoietic transcription factors, we overexpressed specific hematopoietic transcription factors (Gata2, Lmo2, Etv2, ERG, and SCL) at an early stage of hematopoietic differentiation. After overexpression, we generated more CD34+ cells with normal expression level of CD43 and CD31, which are used to define various hematopoietic progenitors. Furthermore, these CD34+ cells possessed normal differentiation potency in colony-forming unit assays and normal gene expression profiles. In this study, we demonstrated that single-cell qPCR can provide guidance for optimization of hematopoietic differentiation and transient overexpression of selected hematopoietic transcription factors can enhance hematopoietic differentiation.

  11. Type I Interferon Receptor Deficiency in Dendritic Cells Facilitates Systemic Murine Norovirus Persistence Despite Enhanced Adaptive Immunity.

    Science.gov (United States)

    Nice, Timothy J; Osborne, Lisa C; Tomov, Vesselin T; Artis, David; Wherry, E John; Virgin, Herbert W

    2016-06-01

    In order for a virus to persist, there must be a balance between viral replication and immune clearance. It is commonly believed that adaptive immunity drives clearance of viral infections and, thus, dysfunction or viral evasion of adaptive immunity is required for a virus to persist. Type I interferons (IFNs) play pleiotropic roles in the antiviral response, including through innate control of viral replication. Murine norovirus (MNoV) replicates in dendritic cells (DCs) and type I IFN signaling in DCs is important for early control of MNoV replication. We show here that the non-persistent MNoV strain CW3 persists systemically when CD11c positive DCs are unable to respond to type I IFN. Persistence in this setting is associated with increased early viral titers, maintenance of DC numbers, increased expression of DC activation markers and an increase in CD8 T cell and antibody responses. Furthermore, CD8 T cell function is maintained during the persistent phase of infection and adaptive immune cells from persistently infected mice are functional when transferred to Rag1-/- recipients. Finally, increased early replication and persistence are also observed in mixed bone marrow chimeras where only half of the CD11c positive DCs are unable to respond to type I IFN. These findings demonstrate that increased early viral replication due to a cell-intrinsic innate immune deficiency is sufficient for persistence and a functional adaptive immune response is not sufficient for viral clearance.

  12. Type I Interferon Receptor Deficiency in Dendritic Cells Facilitates Systemic Murine Norovirus Persistence Despite Enhanced Adaptive Immunity.

    Directory of Open Access Journals (Sweden)

    Timothy J Nice

    2016-06-01

    Full Text Available In order for a virus to persist, there must be a balance between viral replication and immune clearance. It is commonly believed that adaptive immunity drives clearance of viral infections and, thus, dysfunction or viral evasion of adaptive immunity is required for a virus to persist. Type I interferons (IFNs play pleiotropic roles in the antiviral response, including through innate control of viral replication. Murine norovirus (MNoV replicates in dendritic cells (DCs and type I IFN signaling in DCs is important for early control of MNoV replication. We show here that the non-persistent MNoV strain CW3 persists systemically when CD11c positive DCs are unable to respond to type I IFN. Persistence in this setting is associated with increased early viral titers, maintenance of DC numbers, increased expression of DC activation markers and an increase in CD8 T cell and antibody responses. Furthermore, CD8 T cell function is maintained during the persistent phase of infection and adaptive immune cells from persistently infected mice are functional when transferred to Rag1-/- recipients. Finally, increased early replication and persistence are also observed in mixed bone marrow chimeras where only half of the CD11c positive DCs are unable to respond to type I IFN. These findings demonstrate that increased early viral replication due to a cell-intrinsic innate immune deficiency is sufficient for persistence and a functional adaptive immune response is not sufficient for viral clearance.

  13. Strategies to facilitate implementation and sustainability of large system transformations: a case study of a national program for improving quality of care for elderly people.

    Science.gov (United States)

    Nyström, Monica Elisabeth; Strehlenert, Helena; Hansson, Johan; Hasson, Henna

    2014-09-18

    Large-scale change initiatives stimulating change in several organizational systems in the health and social care sector are challenging both to lead and evaluate. There is a lack of systematic research that can enrich our understanding of strategies to facilitate large system transformations in this sector. The purpose of this study was to examine the characteristics of core activities and strategies to facilitate implementation and change of a national program aimed at improving life for the most ill elderly people in Sweden. The program outcomes were also addressed to assess the impact of these strategies. A longitudinal case study design with multiple data collection methods was applied. Archival data (n = 795), interviews with key stakeholders (n = 11) and non-participant observations (n = 23) were analysed using content analysis. Outcome data was obtained from national quality registries. This study presents an approach for implementing a large national change program that is characterized by initial flexibility and dynamism regarding content and facilitation strategies and a growing complexity over time requiring more structure and coordination. The description of activities and strategies show that the program management team engaged a variety of stakeholders and actor groups and accordingly used a palate of different strategies. The main strategies used to influence change in the target organisations were to use regional improvement coaches, regional strategic management teams, national quality registries, financial incentives and annually revised agreements. Interactive learning sessions, intense communication, monitor and measurements, and active involvement of different experts and stakeholders, including elderly people, complemented these strategies. Program outcomes showed steady progress in most of the five target areas, less so for the target of achieving coordinated care. There is no blue-print on how to approach the challenging task of

  14. Public perceptions of animal cloning

    DEFF Research Database (Denmark)

    Jelsøe, Erling; Vincentsen, Ulla; Andersen, Ida-Elisabeth

    What was from the outset meant to be a survey testing predefined categories of ethical positions related to new biotechnologies with animal cloning as an example was subsequently developed into a process of broader involvement of groups of citizens in the issue. The survey was conducted at meetings...... in four different cities in Denmark. The participants were introduced to animal cloning and after that they filled out the questionnaire. Finally, the issue was discussed in focus groups. The process as a whole was run in a dialogue oriented way. Through the information they received in combination...... with reflecting on the survey questions the participants were well prepared for discussions in the focus groups. This approach made it possible, on the one hand to get a measure of the citizen's perceptions of the ethical aspects of animal cloning, but also to go deeper into their own thoughts of the issue...

  15. Whole-system change: case study of factors facilitating early implementation of a primary health care reform in a South African province.

    Science.gov (United States)

    Schneider, Helen; English, Rene; Tabana, Hanani; Padayachee, Thesandree; Orgill, Marsha

    2014-11-29

    Whole-system interventions are those that entail system wide changes in goals, service delivery arrangements and relationships between actors, requiring approaches to implementation that go beyond projects or programmes. Drawing on concepts from complexity theory, this paper describes the catalysts to implementation of a whole-system intervention in the North West Province of South Africa. This province was an early adopter of a national primary health care (PHC) strategy that included the establishment of PHC outreach teams based on generalist community health workers. We interviewed a cross section of provincial actors, from senior to frontline, observed processes and reviewed secondary data, to construct a descriptive-explanatory case study of early implementation of the PHC outreach team strategy and the factors facilitating this in the province. Implementation of the PHC outreach team strategy was characterised by the following features: 1) A favourable provincial context of a well established district and sub-district health system and long standing values in support of PHC; 2) The forging of a collective vision for the new strategy that built on prior history and values and that led to distributed leadership and ownership of the new policy; 3) An implementation strategy that ensured alignment of systems (information, human resources) and appropriate sequencing of activities (planning, training, piloting, household campaigns); 4) The privileging of 'community dialogues' and local manager participation in the early phases; 5) The establishment of special implementation structures: a PHC Task Team (chaired by a senior provincial manager) to enable feedback and ensure accountability, and an NGO partnership that provided flexible support for implementation. These features resonate with the deliberative, multi-level and context sensitive approaches described as the "simple rules" of successful PHC system change in other settings. Although implementation was not

  16. Phage lambda cDNA cloning vectors for subtractive hybridization, fusion-protein synthesis and Cre-loxP automatic plasmid subcloning.

    Science.gov (United States)

    Palazzolo, M J; Hamilton, B A; Ding, D L; Martin, C H; Mead, D A; Mierendorf, R C; Raghavan, K V; Meyerowitz, E M; Lipshitz, H D

    1990-03-30

    We describe the construction and use of two classes of cDNA cloning vectors. The first class comprises the lambda EXLX(+) and lambda EXLX(-) vectors that can be used for the expression in Escherichia coli of proteins encoded by cDNA inserts. This is achieved by the fusion of cDNA open reading frames to the T7 gene 10 promoter and protein-coding sequences. The second class, the lambda SHLX vectors, allows the generation of large amounts of single-stranded DNA or synthetic cRNA that can be used in subtractive hybridization procedures. Both classes of vectors are designed to allow directional cDNA cloning with non-enzymatic protection of internal restriction sites. In addition, they are designed to facilitate conversion from phage lambda to plasmid clones using a genetic method based on the bacteriophage P1 site-specific recombination system; we refer to this as automatic Cre-loxP plasmid subcloning. The phage lambda arms, lambda LOX, used in the construction of these vectors have unique restriction sites positioned between the two loxP sites. Insertion of a specialized plasmid between these sites will convert it into a phage lambda cDNA cloning vector with automatic plasmid subcloning capability.

  17. Integration of an optical coherence tomography (OCT) system into an examination incubator to facilitate in vivo imaging of cardiovascular development in higher vertebrate embryos under stable physiological conditions

    DEFF Research Database (Denmark)

    Happel, Christoph M.; Thrane, Lars; Thommes, Jan

    2011-01-01

    High-resolution in vivo imaging of higher vertebrate embryos over short or long time periods under constant physiological conditions is a technically challenging task for researchers working on cardiovascular development. In chick embryos, for example, various studies have shown that without...... significance, should be documented under physiological conditions. However, previous studies were mostly carried out outside of an incubator or under suboptimal environmental conditions. Here we present, to the best of our knowledge, the first detailed description of an optical coherence tomography (OCT......) system integrated into an examination incubator to facilitate real-time in vivo imaging of cardiovascular development under physiological environmental conditions. We demonstrate the suitability of this OCT examination incubator unit for use in cardiovascular development studies by examples of proof...

  18. Quantum cloning machines and the applications

    International Nuclear Information System (INIS)

    Fan, Heng; Wang, Yi-Nan; Jing, Li; Yue, Jie-Dong; Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu

    2014-01-01

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the security of quantum key distribution protocols such as BB84 protocol, six-state protocol, B92 protocol and their generalizations. Some well-known quantum cloning machines include universal quantum cloning machine, phase-covariant cloning machine, the asymmetric quantum cloning machine and the probabilistic quantum cloning machine. In the past years, much progress has been made in studying quantum cloning machines and their applications and implementations, both theoretically and experimentally. In this review, we will give a complete description of those important developments about quantum cloning and some related topics. On the other hand, this review is self-consistent, and in particular, we try to present some detailed formulations so that further study can be taken based on those results

  19. Healthy ageing of cloned sheep.

    Science.gov (United States)

    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

    2016-07-26

    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.

  20. EasyClone-MarkerFree

    DEFF Research Database (Denmark)

    Fabre, Mathew Malcolm Jessop; Jakociunas, Tadas; Stovicek, Vratislav

    2016-01-01

    Saccharomyces cerevisiae is an established industrial host for production of recombinant proteins, fuels and chemicals. To enable stable integration of multiple marker-free overexpression cassettes in the genome of S. cerevisiae, we have developed a vector toolkit EasyClone-MarkerFree. The integr...... standardized genome engineering, and should be of particular interest to researchers working on yeast chassis with limited markers available....

  1. Clone Poems and the Microcomputer.

    Science.gov (United States)

    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)

  2. Human reproductive cloning: a conflict of liberties.

    Science.gov (United States)

    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.

  3. Probabilistic cloning and deleting of quantum states

    International Nuclear Information System (INIS)

    Feng Yuan; Zhang Shengyu; Ying Mingsheng

    2002-01-01

    We construct a probabilistic cloning and deleting machine which, taking several copies of an input quantum state, can output a linear superposition of multiple cloning and deleting states. Since the machine can perform cloning and deleting in a single unitary evolution, the probabilistic cloning and other cloning machines proposed in the previous literature can be thought of as special cases of our machine. A sufficient and necessary condition for successful cloning and deleting is presented, and it requires that the copies of an arbitrarily presumed number of the input states are linearly independent. This simply generalizes some results for cloning. We also derive an upper bound for the success probability of the cloning and deleting machine

  4. Visual explorer facilitator's guide

    CERN Document Server

    Palus, Charles J

    2010-01-01

    Grounded in research and practice, the Visual Explorer™ Facilitator's Guide provides a method for supporting collaborative, creative conversations about complex issues through the power of images. The guide is available as a component in the Visual Explorer Facilitator's Letter-sized Set, Visual Explorer Facilitator's Post card-sized Set, Visual Explorer Playing Card-sized Set, and is also available as a stand-alone title for purchase to assist multiple tool users in an organization.

  5. Probabilistic cloning of three symmetric states

    International Nuclear Information System (INIS)

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

    2010-01-01

    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.

  6. A single-copy galK promoter cloning vector suitable for cloning strong promoters

    DEFF Research Database (Denmark)

    Dandanell, Gert; Court, Donald L.; Hammer, Karin

    1986-01-01

    We report the construction of lambda galK promoter cloning vectors for cloning and characterization of strong promoters. This phage, which contains a unique HindIII cloning site, was applied to the cloning and analysis of transcription initiations of the regulatory region of the deo-operon of...

  7. The Combinational Use of CRISPR/Cas9 and Targeted Toxin Technology Enables Efficient Isolation of Bi-Allelic Knockout Non-Human Mammalian Clones

    Directory of Open Access Journals (Sweden)

    Satoshi Watanabe

    2018-04-01

    Full Text Available Recent advances in genome editing systems such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9 have facilitated genomic modification in mammalian cells. However, most systems employ transient treatment with selective drugs such as puromycin to obtain the desired genome-edited cells, which often allows some untransfected cells to survive and decreases the efficiency of generating genome-edited cells. Here, we developed a novel targeted toxin-based drug-free selection system for the enrichment of genome-edited cells. Cells were transfected with three expression vectors, each of which carries a guide RNA (gRNA, humanized Cas9 (hCas9 gene, or Clostridium perfringens-derived endo-β-galactosidase C (EndoGalC gene. Once EndoGalC is expressed in a cell, it digests the cell-surface α-Gal epitope, which is specifically recognized by BS-I-B4 lectin (IB4. Three days after transfection, these cells were treated with cytotoxin saporin-conjugated IB4 (IB4SAP for 30 min at 37 °C prior to cultivation in a normal medium. Untransfected cells and those weakly expressing EndoGalC will die due to the internalization of saporin. Cells transiently expressing EndoGalC strongly survive, and some of these surviving clones are expected to be genome-edited bi-allelic knockout (KO clones due to their strong co-expression of gRNA and hCas9. When porcine α-1,3-galactosyltransferase gene, which can synthesize the α-Gal epitope, was attempted to be knocked out, 16.7% and 36.7% of the surviving clones were bi-allelic and mono-allelic knockout (KO cells, respectively, which was in contrast to the isolation of clones in the absence of IB4SAP treatment. Namely, 0% and 13.3% of the resulting clones were bi-allelic and mono-allelic KO cells, respectively. A similar tendency was seen when other target genes such as DiGeorge syndrome critical region gene 2 and transforming growth factor-β receptor type 1 gene were

  8. Entangled cloning of stabilizer codes and free fermions

    Science.gov (United States)

    Hsieh, Timothy H.

    2016-10-01

    Though the no-cloning theorem [Wooters and Zurek, Nature (London) 299, 802 (1982), 10.1038/299802a0] prohibits exact replication of arbitrary quantum states, there are many instances in quantum information processing and entanglement measurement in which a weaker form of cloning may be useful. Here, I provide a construction for generating an "entangled clone" for a particular but rather expansive and rich class of states. Given a stabilizer code or free fermion Hamiltonian, this construction generates an exact entangled clone of the original ground state, in the sense that the entanglement between the original and the exact copy can be tuned to be arbitrarily small but finite, or large, and the relation between the original and the copy can also be modified to some extent. For example, this Rapid Communication focuses on generating time-reversed copies of stabilizer codes and particle-hole transformed ground states of free fermion systems, although untransformed clones can also be generated. The protocol leverages entanglement to simulate a transformed copy of the Hamiltonian without having to physically implement it and can potentially be realized in superconducting qubits or ultracold atomic systems.

  9. Reproductive cloning: past, present and future.

    Science.gov (United States)

    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.

  10. Psychological aspects of human cloning and genetic manipulation: the identity and uniqueness of human beings.

    Science.gov (United States)

    Morales, N M

    2009-01-01

    Human cloning has become one of the most controversial debates about reproduction in Western civilization. Human cloning represents asexual reproduction, but the critics of human cloning argue that the result of cloning is not a new individual who is genetically unique. There is also awareness in the scientific community, including the medical community, that human cloning and the creation of clones are inevitable. Psychology and other social sciences, together with the natural sciences, will need to find ways to help the healthcare system, to be prepared to face the new challenges introduced by the techniques of human cloning. One of those challenges is to help the healthcare system to find specific standards of behaviour that could be used to help potential parents to interact properly with cloned babies or children created through genetic manipulation. In this paper, the concepts of personality, identity and uniqueness are discussed in relationship to the contribution of twin studies in these areas. The author argues that an individual created by human cloning techniques or any other type of genetic manipulation will not show the donor's characteristics to the extent of compromising uniqueness. Therefore, claims to such an effect are needlessly alarmist.

  11. Training facilitators and supervisors

    DEFF Research Database (Denmark)

    Kjær, Louise Binow; O Connor, Maja; Krogh, Kristian

    At the Master’s program in Medicine at Aarhus University, Denmark, we have developed a faculty development program for facilitators and supervisors in 4 progressing student modules in communication, cooperation, and leadership. 1) A course for module 1 and 3 facilitators inspired by the apprentic...

  12. Trade Facilitation in Ethiopia:

    African Journals Online (AJOL)

    Tilahun_EK

    (UNCTAD), “Trade and transport facilitation … addresses a wide agenda in economic development and trade that may include improving transport infrastructure and services, reducing customs tariffs, and removing non-tariff trade barriers including administrative and regulatory barriers.”24. The definition of trade facilitation ...

  13. Learning facilitating leadership

    DEFF Research Database (Denmark)

    Rasmussen, Lauge Baungaard; Hansen, Mette Sanne

    2016-01-01

    deployed for this paper is empirical and conceptual. A specific facilitation project carried out by six international engineering students is presented. The importance of combining cognitive, emotional and synergistic skills is highlighted on the basis of this example, the authors’ extensive experience......This paper explains how engineering students at a Danish university acquired the necessary skills to become emergent facilitators of organisational development. The implications of this approach are discussed and related to relevant viewpoints and findings in the literature. The methodology...... in teaching facilitation and the literature. These types of skills are most effectively acquired by combining conceptual lectures, classroom exercises and the facilitation of groups in a real-life context. The paper also reflects certain ‘shadow sides’ related to facilitation observed by the students...

  14. No-cloning theorem on quantum logics

    International Nuclear Information System (INIS)

    Miyadera, Takayuki; Imai, Hideki

    2009-01-01

    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. Barriers and Facilitators to Online Portal Use Among Patients and Caregivers in a Safety Net Health Care System: A Qualitative Study.

    Science.gov (United States)

    Tieu, Lina; Sarkar, Urmimala; Schillinger, Dean; Ralston, James D; Ratanawongsa, Neda; Pasick, Rena; Lyles, Courtney R

    2015-12-03

    Patient portals have the potential to support self-management for chronic diseases and improve health outcomes. With the rapid rise in adoption of patient portals spurred by meaningful use incentives among safety net health systems (a health system or hospital providing a significant level of care to low-income, uninsured, and vulnerable populations), it is important to understand the readiness and willingness of patients and caregivers in safety net settings to access their personal health records online. To explore patient and caregiver perspectives on online patient portal use before its implementation at San Francisco General Hospital, a safety net hospital. We conducted 16 in-depth interviews with chronic disease patients and caregivers who expressed interest in using the Internet to manage their health. Discussions focused on health care experiences, technology use, and interest in using an online portal to manage health tasks. We used open coding to categorize all the barriers and facilitators to portal use, followed by a second round of coding that compared the categories to previously published findings. In secondary analyses, we also examined specific barriers among 2 subgroups: those with limited health literacy and caregivers. We interviewed 11 patients and 5 caregivers. Patients were predominantly male (82%, 9/11) and African American (45%, 5/11). All patients had been diagnosed with diabetes and the majority had limited health literacy (73%, 8/11). The majority of caregivers were female (80%, 4/5), African American (60%, 3/5), caregivers of individuals with diabetes (60%, 3/5), and had adequate health literacy (60%, 3/5). A total of 88% (14/16) of participants reported interest in using the portal after viewing a prototype. Major perceived barriers included security concerns, lack of technical skills/interest, and preference for in-person communication. Facilitators to portal use included convenience, health monitoring, and improvements in patient

  16. Patient-, organization-, and system-level barriers and facilitators to preventive oral health care: a convergent mixed-methods study in primary dental care.

    Science.gov (United States)

    Templeton, Anna Rose; Young, Linda; Bish, Alison; Gnich, Wendy; Cassie, Heather; Treweek, Shaun; Bonetti, Debbie; Stirling, Douglas; Macpherson, Lorna; McCann, Sharon; Clarkson, Jan; Ramsay, Craig

    2016-01-12

    Dental caries is the most common chronic disease of adult and childhood, a largely preventable yet widespread, costly public health problem. This study identified patient-, organization-, and system-level factors influencing routine delivery of recommended care for prevention and management of caries in primary dental care. A convergent mixed-methods design assessed six guidance-recommended behaviours to prevent and manage caries (recording risk, risk-based recall intervals, applying fluoride varnish, placing preventive fissure sealants, demonstrating oral health maintenance, taking dental x-rays). A diagnostic questionnaire assessing current practice, beliefs, and practice characteristics was sent to a random sample of 651 dentists in National Health Service (NHS) Scotland. Eight in-depth case studies comprising observation of routine dental visits and dental team member interviews were conducted. Patient feedback was collected from adult patients with recent checkups at case study practices. Key informant interviews were conducted with decision makers in policy, funding, education, and regulation. The Theoretical Domains Framework within the Behaviour Change Wheel was used to identify and describe patient-, organization-, and system-level barriers and facilitators to care. Findings were merged into a matrix describing theoretical domains salient to each behaviour. The matrix and Behaviour Change Wheel were used to prioritize behaviours for change and plan relevant intervention strategies. Theoretical domains associated with best practice were identified from the questionnaire (N-196), case studies (N = 8 practices, 29 interviews), and patient feedback (N = 19). Using the study matrix, key stakeholders identified priority behaviours (use of preventive fissure sealants among 6-12-year-olds) and strategies (audit and feedback, patient informational campaign) to improve guidance implementation. Proposed strategies were assessed as appropriate for immediate

  17. Software-Supported USER Cloning Strategies for Site-Directed Mutagenesis and DNA Assembly

    DEFF Research Database (Denmark)

    Genee, Hans Jasper; Bonde, Mads Tvillinggaard; Bagger, Frederik Otzen

    2015-01-01

    USER cloning is a fast and versatile method for engineering of plasmid DNA. We have developed a user friendly Web server tool that automates the design of optimal PCR primers for several distinct USER cloning-based applications. Our Web server, named AMUSER (Automated DNA Modifications with USER...... cloning), facilitates DNA assembly and introduction of virtually any type of site-directed mutagenesis by designing optimal PCR primers for the desired genetic changes. To demonstrate the utility, we designed primers for a simultaneous two-position site-directed mutagenesis of green fluorescent protein...... (GFP) to yellow fluorescent protein (YFP), which in a single step reaction resulted in a 94% cloning efficiency. AMUSER also supports degenerate nucleotide primers, single insert combinatorial assembly, and flexible parameters for PCR amplification. AMUSER is freely available online at ....

  18. Embryo Aggregation in Pig Improves Cloning Efficiency and Embryo Quality.

    Science.gov (United States)

    Buemo, Carla Paola; Gambini, Andrés; Moro, Lucia Natalia; Hiriart, María Inés; Fernández-Martín, Rafael; Collas, Philippe; Salamone, Daniel Felipe

    2016-01-01

    In this study, we analyzed the effects of the cloned embryo aggregation on in vitro embryo development and embryo quality by measuring blastocyst diameter and cell number, DNA fragmentation levels and the expression of genes associated with pluripotency, apoptosis, trophoblast and DNA methylation in the porcine. Zona-free reconstructed cloned embryos were cultured in the well of the well system, placing one (1x non aggregated group) or three (3x group) embryos per microwell. Our results showed that aggregation of three embryos increased blastocyst formation rate and blastocyst diameter of cloned pig embryos. DNA fragmentation levels in 3x aggregated cloned blastocysts were significantly decreased compared to 1x blastocysts. Levels of Oct4, Klf4, Igf2, Bax and Dnmt 1 transcripts were significantly higher in aggregated embryos, whereas Nanog levels were not affected. Transcripts of Cdx2 and Bcl-xl were essentially non-detectable. Our study suggests that embryo aggregation in the porcine may be beneficial for cloned embryo development and embryo quality, through a reduction in apoptotic levels and an improvement in cell reprogramming.

  19. El envejecimiento de los clones

    OpenAIRE

    Trippi, Victorio S.

    2007-01-01

    El envejecimiento de los clones se observa en plantas que muestran crecimiento definido por un determinismo genético, cuando se multiplican con tejidos que evolucionan hacia el crecimiento reproductivo. Las plantas fuertemente influenciadas por el ambiente, pueden mostrar fenómenos de senescencia cuando la condición de ambiente determina el crecimiento reproductivo. Los cambios asociados con la edad resultan de alteraciones del citoplasma como un tipo de diferenciación cel...

  20. Cloning Expeditions: Risky but Rewarding

    Science.gov (United States)

    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

  1. Molecular cloning of cellulase genes from indigenous bacterial isolates

    International Nuclear Information System (INIS)

    Jong Bor Chyan; Pauline Liew Woan Ying; Mat Rasol Awang

    2006-01-01

    Indigenous cellulolytic bacterial isolates having high activities in degrading carboxymethyl cellulose (CMC) were isolated from local environments. Identification of these isolates were performed by molecular techniques. By using polymerase chain reaction (PCR) techniques, PCR products encoding cellulase gene were amplified from the total genomic DNAs. Purified PCR product was successfully cloned and expressed in Escherichia coli host system. The complete nucleotide sequences of the cellulase genes determined. The analysis of amino acid sequences deduced from the genes indicated that the cloned DNA fragments show high homology to those of endoglucanase genes of family GH5. All cloned genes consist of an N-terminal signal peptide, a catalytic domain of family 5 glycosyl hydrolase and a cellulose-binding domain of family III. (Author)

  2. MOLECULAR CLONING OF OVINE cDNA LEPTIN GENE

    Directory of Open Access Journals (Sweden)

    CLAUDIA TEREZIA SOCOL

    2008-05-01

    Full Text Available An efficient bacterial transformation system suitable for cloning the coding sequence of the ovine leptin gene in E. coli DH5α host cells using the pGEMT easy vector it is described in this paper. The necessity of producing leptin is based on the fact that the role of this molecule in the animal and human organism is still unknown, leptin not existing as commercial product on the Romanian market. The results obtained in the bacterial transformation, cloning, recombinant clones selection, control of the insertion experiments and DNA computational analysis represent the first steps in further genetic engineering experiments such as production of DNA libraries, DNA sequencing, protein expression, etc., for a further contribution in elucidating the role of leptin in the animal and human organism.

  3. Low-Flow Extracorporeal Carbon Dioxide Removal Using the Hemolung Respiratory Dialysis System® to Facilitate Lung-Protective Mechanical Ventilation in Acute Respiratory Distress Syndrome.

    Science.gov (United States)

    Akkanti, Bindu; Rajagopal, Keshava; Patel, Kirti P; Aravind, Sangeeta; Nunez-Centanu, Emmanuel; Hussain, Rahat; Shabari, Farshad Raissi; Hofstetter, Wayne L; Vaporciyan, Ara A; Banjac, Igor S; Kar, Biswajit; Gregoric, Igor D; Loyalka, Pranav

    2017-06-01

    Extracorporeal carbon dioxide removal (ECCO 2 R) permits reductions in alveolar ventilation requirements that the lungs would otherwise have to provide. This concept was applied to a case of hypercapnia refractory to high-level invasive mechanical ventilator support. We present a case of an 18-year-old man who developed post-pneumonectomy acute respiratory distress syndrome (ARDS) after resection of a mediastinal germ cell tumor involving the left lung hilum. Hypercapnia and hypoxemia persisted despite ventilator support even at traumatic levels. ECCO 2 R using a miniaturized system was instituted and provided effective carbon dioxide elimination. This facilitated establishment of lung-protective ventilator settings and lung function recovery. Extracorporeal lung support increasingly is being applied to treat ARDS. However, conventional extracorporeal membrane oxygenation (ECMO) generally involves using large cannulae capable of carrying high flow rates. A subset of patients with ARDS has mixed hypercapnia and hypoxemia despite high-level ventilator support. In the absence of profound hypoxemia, ECCO 2 R may be used to reduce ventilator support requirements to lung-protective levels, while avoiding risks associated with conventional ECMO.

  4. Highly Efficient CRISPR/Cas9-Mediated Cloning and Functional Characterization of Gastric Cancer-Derived Epstein-Barr Virus Strains.

    Science.gov (United States)

    Kanda, Teru; Furuse, Yuki; Oshitani, Hitoshi; Kiyono, Tohru

    2016-05-01

    The Epstein-Barr virus (EBV) is etiologically linked to approximately 10% of gastric cancers, in which viral genomes are maintained as multicopy episomes. EBV-positive gastric cancer cells are incompetent for progeny virus production, making viral DNA cloning extremely difficult. Here we describe a highly efficient strategy for obtaining bacterial artificial chromosome (BAC) clones of EBV episomes by utilizing a CRISPR/Cas9-mediated strand break of the viral genome and subsequent homology-directed repair. EBV strains maintained in two gastric cancer cell lines (SNU719 and YCCEL1) were cloned, and their complete viral genome sequences were determined. Infectious viruses of gastric cancer cell-derived EBVs were reconstituted, and the viruses established stable latent infections in immortalized keratinocytes. While Ras oncoprotein overexpression caused massive vacuolar degeneration and cell death in control keratinocytes, EBV-infected keratinocytes survived in the presence of Ras expression. These results implicate EBV infection in predisposing epithelial cells to malignant transformation by inducing resistance to oncogene-induced cell death. Recent progress in DNA-sequencing technology has accelerated EBV whole-genome sequencing, and the repertoire of sequenced EBV genomes is increasing progressively. Accordingly, the presence of EBV variant strains that may be relevant to EBV-associated diseases has begun to attract interest. Clearly, the determination of additional disease-associated viral genome sequences will facilitate the identification of any disease-specific EBV variants. We found that CRISPR/Cas9-mediated cleavage of EBV episomal DNA enabled the cloning of disease-associated viral strains with unprecedented efficiency. As a proof of concept, two gastric cancer cell-derived EBV strains were cloned, and the infection of epithelial cells with reconstituted viruses provided important clues about the mechanism of EBV-mediated epithelial carcinogenesis. This

  5. Understanding text as social practice: An exploration of the potential of systemic functional grammar to facilitate students' interpretation of media texts.

    Directory of Open Access Journals (Sweden)

    Jenny Clarence-Fincham

    2008-08-01

    Full Text Available It has frequently been claimed that Halliday's Systemic Functional Grammar (SFG is apowerful linguistic tool which facilitates analytical and interpretative skills and provides aflexible, yet structured set of analytical tools with which to interpret texts. With this claim asa backdrop, this article asks whether SFG is, in fact an appropriate analytical approach forunder-graduate students and whether it can facilitate their ability to analyse texts. Its contextis a second level course, Analysing Media Texts, offered at Natal University. Broadly framedby critical discourse analysis, it traces the development of a thirteen week module and,using student analyses for illustrative purposes, identifies pedagogical challenges anddifficulties that need to be confronted before any strong claims can be made. It is concludedthat, on the evidence of students' responses to texts analysed during this course, it is not yetpossible to make strong claims about the benefits of SFG. There is enough positiveevidence, however, to pursue the possibility that with innovative curriculum development andthe careful scaffolding and integration of concepts, SFG will be clearly shown to have anextremely important role to play.Daar is dikwels beweer dat Halliday se Sistemies-Funksionele Grammatika (SFG 'n kragtige linguistiese middel is wat analitiese en interpreterende vaardighede bevorder en 'n plooibare, dog gestruktureere stel analitiese gereedskap verskaf waarmee tekste gei"nterpreteer kan word. Met die bewering as agtergrond vra hierdie artikel of SFG inderdaad 'n toepas like analitiese benadering vir voorgraadse studente is en of dit hulle vermoe om tekste te ontleed, bevorder. Die konteks is 'n tweedejaarskursus, Analysing Media Texts, wat aan die Universiteit van Natal aangebied word. Breedweg omraam deur kritiese diskoersanalise, speur die artikel die ontwikkeling van 'n module van dertien weke na, met gebruik van studenteontledings ter illustrasie en identifiseer

  6. resistance of napier grass clones to napier grass stunt disease

    African Journals Online (AJOL)

    ACSS

    Napier grass (Pennisetum purpureum Schumach) is the major livestock fodder under intensive and semi-intensive systems in East Africa. However, the productivity of the grass is constrained by Napier grass Stunt Disease. (NSD). The purpose of this study was to identify Napier grass clones with resistance to NSD.

  7. cDNA cloning and polymorphic domains of the major ...

    African Journals Online (AJOL)

    Major histocompatibility complex (MHC) is a highly polymorphic gene and plays an important role in immune system for vertebrate. To understand the polymorphism character of domestic, we cloned 32 cDNAs of MHC class I α genes of two local chicken breeds in different areas of China. There were 112 variable amino ...

  8. Resistance of Napier grass clones to Napier grass Stunt Disease ...

    African Journals Online (AJOL)

    Napier grass (Pennisetum purpureum Schumach) is the major livestock fodder under intensive and semi-intensive systems in East Africa. However, the productivity of the grass is constrained by Napier grass Stunt Disease (NSD). The purpose of this study was to identify Napier grass clones with resistance to NSD.

  9. cDNA cloning and polymorphic domains of the major ...

    African Journals Online (AJOL)

    Jane

    2011-10-24

    Oct 24, 2011 ... Major histocompatibility complex (MHC) is a highly polymorphic gene and plays an important role in immune system for vertebrate. To understand the polymorphism character of domestic, we cloned 32. cDNAs of MHC class I α genes of two local chicken breeds in different areas of China. There were 112.

  10. Cloning of a gene encoding glycosyltransferase from Pueraria lobata

    African Journals Online (AJOL)

    user

    2011-01-03

    Jan 3, 2011 ... These results suggest that, the PlUGT1 protein can be expressed efficiently in the P. pastoris expression system and may supply a new economic and convenient way for the production of PlUGT1 protein. Key words: Pueraria lobata (Willd.) Ohwi, glycosyltransferase, cloning, expression, Pichia pastoris.

  11. Service user and care giver involvement in mental health system strengthening in Nepal: a qualitative study on barriers and facilitating factors.

    Science.gov (United States)

    Gurung, Dristy; Upadhyaya, Nawaraj; Magar, Jananee; Giri, Nir Prakash; Hanlon, Charlotte; Jordans, Mark J D

    2017-01-01

    Service user and caregiver involvement has become an increasingly common strategy to enhance mental health outcomes, and has been incorporated in the mental healthpolicies of many developed nations. However, this practice is non-existent or fragmented in low and middle income countries (LMICs). Instances of service user and caregiver involvement have been rising slowly in a few LMICs, but are rarely described in the literature. Very little is known about the context of user and caregiver participation in mental health system strengthening processes in a low-income, disaster- and conflict-affected state such as Nepal. This study explores (a) the extent and experiences of service user and caregiver involvement in policy making, service planning, monitoring, and research in Nepal; (b) perceived barriers to such involvement; and (c) possible strategies to overcome barriers. Key Informant Interviews (n = 24) were conducted with service users and caregivers who were either affiliated to a mental health organization or receiving menta health care integrated within primary care. Purposive sampling was employed. Data collection was carried out in 2014 in Chitwan and Kathmandu districts of Nepal. Data analysis was carried out in NVivo10 using a framework approach. The involvement of service users affiliated to mental health organizations in policy development was reported to be 'tokenistic'. Involvement of caregivers was non-existent. Perceived barriers to greater involvement included lack of awareness, stigma and discrimination, poor economic conditions, the centralized health system, and lack of strong leadership and unity among user organizations. Increased focus on reducing public as well as self-stigma, improved policy frameworks and initiatives, and decentralization of care are some strategies that may facilitate service user and caregiver involvement. The study highlighted need for user and caregiver networks free from competing interests and priorities. Improved

  12. Facilitating Knowledge Sharing

    DEFF Research Database (Denmark)

    Holdt Christensen, Peter

    Abstract This paper argues that knowledge sharing can be conceptualized as different situations of exchange in which individuals relate to each other in different ways, involving different rules, norms and traditions of reciprocity regulating the exchange. The main challenge for facilitating...... and the intermediaries regulating the exchange, and facilitating knowledge sharing should therefore be viewed as a continuum of practices under the influence of opportunistic behaviour, obedience or organizational citizenship behaviour. Keywords: Knowledge sharing, motivation, organizational settings, situations...

  13. Hybrid sequencing approach applied to human fecal metagenomic clone libraries revealed clones with potential biotechnological applications.

    Directory of Open Access Journals (Sweden)

    Mária Džunková

    Full Text Available Natural environments represent an incredible source of microbial genetic diversity. Discovery of novel biomolecules involves biotechnological methods that often require the design and implementation of biochemical assays to screen clone libraries. However, when an assay is applied to thousands of clones, one may eventually end up with very few positive clones which, in most of the cases, have to be "domesticated" for downstream characterization and application, and this makes screening both laborious and expensive. The negative clones, which are not considered by the selected assay, may also have biotechnological potential; however, unfortunately they would remain unexplored. Knowledge of the clone sequences provides important clues about potential biotechnological application of the clones in the library; however, the sequencing of clones one-by-one would be very time-consuming and expensive. In this study, we characterized the first metagenomic clone library from the feces of a healthy human volunteer, using a method based on 454 pyrosequencing coupled with a clone-by-clone Sanger end-sequencing. Instead of whole individual clone sequencing, we sequenced 358 clones in a pool. The medium-large insert (7-15 kb cloning strategy allowed us to assemble these clones correctly, and to assign the clone ends to maintain the link between the position of a living clone in the library and the annotated contig from the 454 assembly. Finally, we found several open reading frames (ORFs with previously described potential medical application. The proposed approach allows planning ad-hoc biochemical assays for the clones of interest, and the appropriate sub-cloning strategy for gene expression in suitable vectors/hosts.

  14. Innate immune responses to obesity in cloned and wild-type domestic pig

    DEFF Research Database (Denmark)

    Højbøge, Tina Rødgaard; Skovgaard, Kerstin; Stagsted, Jan

    months of age. mRNA expression levels were determined for 39 innate immune factors on a high-throughput qPCR system in samples from liver, abdominal fat, mesenteric fat and subcutaneous fat. Previous findings have suggested that cloning may affect certain phenotypic traits of pigs including basic...... concentrations and responsiveness of components of the innate immune system. Terminal body weights at 7½ - 9½ months of age were significantly higher for both (WT and cloned) obese groups compared to the lean groups. However, obese WT pigs weighed significantly more than obese cloned pigs (P... significant differences between WT and cloned pigs in the gene response to obesity. Thus, significant phenotypic differences were established for central innate immune factors between cloned and WT pigs, including differences in the response of these factors to an obesity-promoting diet. This should be taken...

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

    Science.gov (United States)

    Marques, M Carmen; Alonso-Cantabrana, Hugo; Forment, Javier; Arribas, Raquel; Alamar, Santiago; Conejero, Vicente; Perez-Amador, Miguel A

    2009-01-01

    Background Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. Indeed, nearly 40% of genes in plants encode proteins of unknown function. Functional characterization of these genes is one of the main challenges in modern biology. In this regard, the availability of full-length cDNA clones may fill in the gap created between sequence information and biological knowledge. Full-length cDNA clones facilitate functional analysis of the corresponding genes enabling manipulation of their expression in heterologous systems and the generation of a variety of tagged versions of the native protein. In addition, the development of full-length cDNA sequences has the power to improve the quality of genome annotation. Results We developed an integrated method to generate a new normalized EST collection enriched in full-length and rare transcripts of different citrus species from multiple tissues and developmental stages. We constructed a total of 15 cDNA libraries, from which we isolated 10,898 high-quality ESTs representing 6142 different genes. Percentages of redundancy and proportion of full-length clones range from 8 to 33, and 67 to 85, respectively, indicating good efficiency of the approach employed. The new EST collection adds 2113 new citrus ESTs, representing 1831 unigenes, to the collection of citrus genes available in the public databases. To facilitate functional analysis, cDNAs were introduced in a Gateway-based cloning vector for high-throughput functional analysis of genes in planta. Herein, we describe the technical methods used in the library construction, sequence analysis of clones and the overexpression of CitrSEP, a citrus homolog to the Arabidopsis SEP3 gene, in Arabidopsis as an example of a practical application of the engineered Gateway vector for functional analysis. Conclusion The new EST collection denotes an

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

    Directory of Open Access Journals (Sweden)

    Alamar Santiago

    2009-09-01

    Full Text Available Abstract Background Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. Indeed, nearly 40% of genes in plants encode proteins of unknown function. Functional characterization of these genes is one of the main challenges in modern biology. In this regard, the availability of full-length cDNA clones may fill in the gap created between sequence information and biological knowledge. Full-length cDNA clones facilitate functional analysis of the corresponding genes enabling manipulation of their expression in heterologous systems and the generation of a variety of tagged versions of the native protein. In addition, the development of full-length cDNA sequences has the power to improve the quality of genome annotation. Results We developed an integrated method to generate a new normalized EST collection enriched in full-length and rare transcripts of different citrus species from multiple tissues and developmental stages. We constructed a total of 15 cDNA libraries, from which we isolated 10,898 high-quality ESTs representing 6142 different genes. Percentages of redundancy and proportion of full-length clones range from 8 to 33, and 67 to 85, respectively, indicating good efficiency of the approach employed. The new EST collection adds 2113 new citrus ESTs, representing 1831 unigenes, to the collection of citrus genes available in the public databases. To facilitate functional analysis, cDNAs were introduced in a Gateway-based cloning vector for high-throughput functional analysis of genes in planta. Herein, we describe the technical methods used in the library construction, sequence analysis of clones and the overexpression of CitrSEP, a citrus homolog to the Arabidopsis SEP3 gene, in Arabidopsis as an example of a practical application of the engineered Gateway vector for functional analysis. Conclusion The new

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

    Science.gov (United States)

    Marques, M Carmen; Alonso-Cantabrana, Hugo; Forment, Javier; Arribas, Raquel; Alamar, Santiago; Conejero, Vicente; Perez-Amador, Miguel A

    2009-09-11

    Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. Indeed, nearly 40% of genes in plants encode proteins of unknown function. Functional characterization of these genes is one of the main challenges in modern biology. In this regard, the availability of full-length cDNA clones may fill in the gap created between sequence information and biological knowledge. Full-length cDNA clones facilitate functional analysis of the corresponding genes enabling manipulation of their expression in heterologous systems and the generation of a variety of tagged versions of the native protein. In addition, the development of full-length cDNA sequences has the power to improve the quality of genome annotation. We developed an integrated method to generate a new normalized EST collection enriched in full-length and rare transcripts of different citrus species from multiple tissues and developmental stages. We constructed a total of 15 cDNA libraries, from which we isolated 10,898 high-quality ESTs representing 6142 different genes. Percentages of redundancy and proportion of full-length clones range from 8 to 33, and 67 to 85, respectively, indicating good efficiency of the approach employed. The new EST collection adds 2113 new citrus ESTs, representing 1831 unigenes, to the collection of citrus genes available in the public databases. To facilitate functional analysis, cDNAs were introduced in a Gateway-based cloning vector for high-throughput functional analysis of genes in planta. Herein, we describe the technical methods used in the library construction, sequence analysis of clones and the overexpression of CitrSEP, a citrus homolog to the Arabidopsis SEP3 gene, in Arabidopsis as an example of a practical application of the engineered Gateway vector for functional analysis. The new EST collection denotes an important step towards the

  18. Cloning and identification of Group 1 mrp operon encoding a novel monovalent cation/proton antiporter system from the moderate halophile Halomonas zhaodongensis.

    Science.gov (United States)

    Meng, Lin; Hong, Shan; Liu, Henan; Huang, Haipeng; Sun, Hao; Xu, Tong; Jiang, Juquan

    2014-11-01

    The novel species Halomonas zhaodongensis NEAU-ST10-25(T) recently identified by our group is a moderate halophile which can grow at the range of 0-2.5 M NaCl (optimum 0.5 M) and pH 6-12 (optimum pH 9). To explore its halo-alkaline tolerant mechanism, genomic DNA was screened from NEAU-ST10-25(T) in this study for Na(+)(Li(+))/H(+) antiporter genes by selection in Escherichia coli KNabc lacking three major Na(+)(Li(+))/H(+) antiporters. One mrp operon could confer tolerance of E. coli KNabc to 0.8 M NaCl and 100 mM LiCl, and an alkaline pH. This operon was previously mainly designated mrp (also mnh, pha or sha) due to its multiple resistance and pH-related activity. Here, we will also use mrp to designate the homolog from H. zhaodongensis (Hz_mrp). Sequence analysis and protein alignment showed that Hz_mrp should belong to Group 1 mrp operons. Further phylogenetic analysis reveals that Hz_Mrp system should represent a novel sub-class of Group 1 Mrp systems. This was confirmed by a significant difference in pH-dependent activity profile or the specificity and affinity for the transported monovalent cations between Hz_Mrp system and all the known Mrp systems. Therefore, we propose that Hz_Mrp should be categorized as a novel Group 1 Mrp system.

  19. N-P-K balance in a milk production system on a C. nlemfuensis grassland and a biomass bank of P. purpureum CT-115 clone

    Science.gov (United States)

    Crespo, G.; Rodriguez, I.; Martinez, O.

    2009-04-01

    In very intensive milk production systems in Europe and America with the use of high amounts of chemical fertilizers, the nutrient recycling models consider the losses by leaching and N volatilization, as well as the hydro physical characteristics of the soil affecting the performance of this element (10; 6). However, in more extensive milk production systems, low input agriculture forming the natural cycle occurring within each farm, is of vital importance to potentate nutrient recycling for a stable animal production. The objective is the determination of the values of N, P and K inputs and outputs in a dairy farm with a sward composed by 60% of C. nlemfuensis and 40% of P. purpureum CT-115, associated with legumes in 28% of the area and the balance of these nutrients in the system using the "Recycling" software proposed by Crespo et al (2007). The grassland covered an area of 53.4 ha, composed by C. nlemfuensis (60%), P. purpureum CT-115 (40%) and L. leucocephala and C. cajan legumes intercropped in 28% of the area. The dairy herd consisted of 114 cows, 35 replacement heifers and 24 calves. There was a milk yield of 100 000 litters and the animals consumed 825 t DM from pastures and 75.1 t DM from other supplementary feeds. Nutrients extracted by pastures, nutrients intake by animals from pastures, symbiotically N fixation by legumes and N, P and K determinations outside the system due to animal production were determined (3-11). Volatilized ammonia, nutrient input and litter accumulated in the paddocks were measured once each season of the year. In the whole system the balance indicates negative values of N, P and K. Out of the total amount of nutrients consumed, animals used only 16 kg N, 5 Kg P and 4 Kg K for milk production, LW gain and calf production, the remainder returned to the system through excretions. Hence, more than 90% of the N and K, and approximately 81% of the P consumed by the animals were recycled to the system through the excretions. These

  20. "FACILS 2014: Microbially-driven facilitation systems in environmental biotechnology" (hereafter "FACILS") presented here by the European Commission (EC)-United States (US) Task Force on Biotechnology Research

    Energy Technology Data Exchange (ETDEWEB)

    Methe, Barbara

    2016-02-03

    As we enter the 21st century, the sustainability of the biosphere is a global challenge that can best be met with a global response. This includes how we train and promote our next generation of research scientists in the emerging arenas of genome-enabled biology and a bio-based economy. It is this fundamental issue that formed the motivation for designing and conducting a shortcourse entitled “FACILIS 2014: Microbially-driven facilitation systems in environmental biotechnology” (hereafter “FACILIS”) presented here by the European Commission (EC)-United States (US) Task Force on Biotechnology Research. This WG was established in 1994 under the umbrella of the US-EC Task Force on Biotechnology Research, a transatlantic collaborative group overseen by the US Office of Science and Technology Policy (OSTP) and the EC. The Environmental Biotechnology Working Group maintains several goals, including establishing research links between scientists in EU countries and the US and fostering the careers of junior scientists from both sides of the Atlantic to the global nature of scientific cooperation. To that end, a shortcourse was held at the University of Milan in Italy on July 12-25 2014 organized around cross-cutting themes of genomic science and designed to attract a stellar group of interdisciplinary early carrier researchers. A total of 22 students, 10 from the US and 12 from the EU participated. The course provided them with hands-on experience with the latest scientific methods in genomics and bioinformatics; using a format that combines lectures, laboratory research and field work with the final goal to enable researchers to finally turn data into knowledge.

  1. A study of the blood flow restriction pressure of a tourniquet system to facilitate development of a system that can prevent musculoskeletal complications.

    Science.gov (United States)

    Maeda, Hiroyuki; Iwase, Hideaki; Kanda, Akio; Morohashi, Itaru; Kaneko, Kazuo; Maeda, Mutsuhiro; Kakinuma, Yuki; Takei, Yusuke; Amemiya, Shota; Mitsui, Kazuyuki

    2017-01-01

    After an emergency or disaster, subsequent trauma can cause severe bleeding and this can often prove fatal, so promptly stopping that bleeding is crucial to preventing avoidable trauma deaths. A tourniquet is often used to restrict blood flow to an extremity. In operation and hospital, the tourniquet systems currently in use are pneumatically actuated by an air compressor, so they must have a steady power supply. These devices have several drawbacks: they vibrate and are noisy since they are pneumatically actuated and they are far from portable since they are large and heavy. Presumably, the drawbacks of pneumatic tourniquets could be overcome by developing a small, lightweight, vibration-free, quiet, and battery-powered tourniquet system. The current study built a small, vibration-free electrohydrodynamic (EHD) pump and then used that pump to restrict blood flow to the leg of rats in an experiment. This study explored the optimal conditions for effective restriction of blood flow by assessing biochemical and musculoskeletal complications following the restriction of blood flow, and this study also examined whether or not an EHD pump could be used to actuate a tourniquet system. A tourniquet cuff (width 12 mm × length 150 mm, material: polyolefin) was placed on the thigh of Wistar rats and pressure was applied for 2 hours by a device that uses EHD phenomena to generate pressure (an EHD pump). Animals were divided into four groups based on how much compressive pressure was applied with a tourniquet: 40 kPa (300 mm Hg, n = 13), 30 kPa (225 mm Hg, n = 12), 20 kPa (150 mm Hg, n = 15), or 0 kPa (controls, n = 25). Tissue oxygen saturation (regional oxygen saturation, denoted here as rSO 2 ) was measured to assess the restriction of blood flow. To assess behavior once blood flow resumed, animal activity was monitored for third day and the amount of movement was counted with digital counters. Body weight was measured before and after the behavioral experiment, and

  2. Experimental Demonstration of Probabilistic Quantum Cloning

    Science.gov (United States)

    Chen, Hongwei; Lu, Dawei; Chong, Bo; Qin, Gan; Zhou, Xianyi; Peng, Xinhua; Du, Jiangfeng

    2011-05-01

    The method of quantum cloning is divided into two main categories: approximate and probabilistic quantum cloning. The former method is used to approximate an unknown quantum state deterministically, and the latter can be used to faithfully copy the state probabilistically. Thus far, many approximate cloning machines have been experimentally demonstrated, but probabilistic cloning remains an experimental challenge, as it requires more complicated networks and a higher level of precision control. In this work, we design an efficient quantum network with a limited amount of resources and perform the first experimental demonstration of probabilistic quantum cloning in a NMR quantum computer. In our experiment, the optimal cloning efficiency proposed by Duan and Guo [Phys. Rev. Lett. 80, 4999 (1998)PRLTAO0031-900710.1103/PhysRevLett.80.4999] is achieved.

  3. Effective and efficient model clone detection

    DEFF Research Database (Denmark)

    Störrle, Harald

    2015-01-01

    Code clones are a major source of software defects. Thus, it is likely that model clones (i.e., duplicate fragments of models) have a significant negative impact on model quality, and thus, on any software created based on those models, irrespective of whether the software is generated fully...... automatically (“MDD-style”) or hand-crafted following the blueprint defined by the model (“MBSD-style”). Unfortunately, however, model clones are much less well studied than code clones. In this paper, we present a clone detection algorithm for UML domain models. Our approach covers a much greater variety...... of model types than existing approaches while providing high clone detection rates at high speed....

  4. Novel cloning machine with supplementary information

    International Nuclear Information System (INIS)

    Qiu Daowen

    2006-01-01

    Probabilistic cloning was first proposed by Duan and Guo. Then Pati established a novel cloning machine (NCM) for copying superposition of multiple clones simultaneously. In this paper, we deal with the novel cloning machine with supplementary information (NCMSI). For the case of cloning two states, we demonstrate that the optimal efficiency of the NCMSI in which the original party and the supplementary party can perform quantum communication equals that achieved by a two-step cloning protocol wherein classical communication is only allowed between the original and the supplementary parties. From this equivalence, it follows that NCMSI may increase the success probabilities for copying. Also, an upper bound on the unambiguous discrimination of two nonorthogonal pure product states is derived. Our investigation generalizes and completes the results in the literature

  5. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2013-01-01

    Code clones (i.e., duplicate fragments of code) have been studied for long, and there is strong evidence that they are a major source of software faults. Anecdotal evidence suggests that this phenomenon occurs similarly in models, suggesting that model clones are as detrimental to model quality...... as they are to code quality. However, programming language code and visual models have significant differences that make it difficult to directly transfer notions and algorithms developed in the code clone arena to model clones. In this article, we develop and propose a definition of the notion of “model clone” based...... on the thorough analysis of practical scenarios. We propose a formal definition of model clones, specify a clone detection algorithm for UML domain models, and implement it prototypically. We investigate different similarity heuristics to be used in the algorithm, and report the performance of our approach. While...

  6. The cloning of T lymphocytes.

    Science.gov (United States)

    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. Copyright © 1982. Published by Elsevier B.V.

  7. No human cloning: a social ethics perspective.

    Science.gov (United States)

    Cahill, L S

    1999-01-01

    This Essay addresses the negative impact of human cloning on the family, and argues further that market incentives to develop and implement cloning techniques exploit and exacerbate socioeconomic inequities. It suggests that cloning should be prohibited internationally and examines possible routes to that aim. To begin with, it offers some reflections on the nature of moral argument, and on the role of religion in public debate.

  8. Experimental Quantum Cloning with Continuous Variables

    Science.gov (United States)

    Andersen, Ulrik L.; Josse, Vincent; Lütkenhaus, Norbert; Leuchs, Gerd

    In this chapter we present a scheme for optimal Gaussian cloning of optical coherent states. Its optical realization is based entirely on simple linear optical elements and homodyne detection. This is in contrast to previous proposals where parametric processes were suggested to be used for optimal Gaussian cloning. The optimality of the presented scheme is only limited by detection inefficiencies. Experimentally we achieved a cloning fidelity of up to 65%, which almost touches the optimal value of 2/3.

  9. Species-specific challenges in dog cloning.

    Science.gov (United States)

    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. © 2012 Blackwell Verlag GmbH.

  10. Human cloning: Eastern Mediterranean Region perspective.

    Science.gov (United States)

    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.

  11. From Teaching to Facilitation

    DEFF Research Database (Denmark)

    de Graaff, Erik

    2013-01-01

    A shift from teaching to learning is characteristic of the introduction of Problem Based Learning (PBL) in an existing school. As a consequence the teaching staff has to be trained in skills like facilitating group work and writing cases. Most importantly a change in thinking about teaching...... and learning will have to be realized. In the implementation of PBL it makes a difference how the core features of the problem and the role of the facilitator have been defined. This paper will present components of a PBL faculty-development training programme and discuss the relevance with respect...

  12. Coal export facilitation

    International Nuclear Information System (INIS)

    Eeles, L.

    1998-01-01

    There is a wide range of trade barriers, particularly tariffs, in current and potential coal market. Commonwealth departments in Australia play a crucial role in supporting government industry policies. This article summarises some of more recent activities of the Department of Primary Industries and Energy (DPIE) in facilitating the export of Australian Coals. Coal export facilitation activities are designed to assist the Australian coal industry by directing Commonwealth Government resources towards issues which would be inappropriate or difficult for the industry to address itself

  13. Cloning human DNA repair genes

    International Nuclear Information System (INIS)

    Jeggo, P.A.; Carr, A.M.; Lehmann, A.R.

    1994-01-01

    Many human genes involved in the repair of UV damage have been cloned using different procedures and they have been of great value in assisting the understanding of the mechanism of nucleotide excision-repair. Genes involved in repair of ionizing radiation damage have proved more difficult to isolate. Positional cloning has localized the XRCC5 gene to a small region of chromosome 2q33-35, and a series of yeast artificial chromosomes covering this region have been isolated. Very recent work has shown that the XRCC5 gene encodes the 80 kDa subunit of the Ku DNA-binding protein. The Ku80 gene also maps to this region. Studies with fission yeast have shown that radiation sensitivity can result not only from defective DNA repair but also from abnormal cell cycle control following DNA damage. Several genes involved in this 'check-point' control in fission yeast have been isolated and characterized in detail. It is likely that a similar checkpoint control mechanism exists in human cells. (author)

  14. High-Throughput Cloning and Expression Library Creation for Functional Proteomics

    Science.gov (United States)

    Festa, Fernanda; Steel, Jason; Bian, Xiaofang; Labaer, Joshua

    2013-01-01

    The study of protein function usually requires the use of a cloned version of the gene for protein expression and functional assays. This strategy is particular important when the information available regarding function is limited. The functional characterization of the thousands of newly identified proteins revealed by genomics requires faster methods than traditional single gene experiments, creating the need for fast, flexible and reliable cloning systems. These collections of open reading frame (ORF) clones can be coupled with high-throughput proteomics platforms, such as protein microarrays and cell-based assays, to answer biological questions. In this tutorial we provide the background for DNA cloning, discuss the major high-throughput cloning systems (Gateway® Technology, Flexi® Vector Systems, and Creator™ DNA Cloning System) and compare them side-by-side. We also report an example of high-throughput cloning study and its application in functional proteomics. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP12). Details can be found at http://www.proteomicstutorials.org. PMID:23457047

  15. [Gene cloning, selection of plasmids and application of Fasciola hepatica cathepsin L1 gene].

    Science.gov (United States)

    Kuk, Salih; Erensoy, Ahmet

    2008-01-01

    Gene cloning refers to the process by which a fragment of DNA is transferred from one organism to a vector. A vector is an agent that can carry a DNA fragment into a host cell. Commonly used vectors include plasmids, lambda phage, cosmid and yeast artificial chromosome (YAC). Plasmid vectors that have been extensively used in genetic engineering are derived from natural plasmids. These contain a genetic marker conferring a phenotype that can be selected for or against and a polylinker or multiple cloning site (MCS), which is a short region containing several commonly used restriction sites allowing the insertion of DNA fragments at this location easily. There are several plasmids and gene cloning kits available nowadays commercially. These kits contain an advanced positive selection system for the highest efficiency cloning of PCR products generated with any DNA polymerase. The kits offer cloning efficiencies of up to 100% positive clones, eliminating the need for tedious colony screening. We consider that researchers should choose the most suitable plasmids and cloning kits for gene cloning in view of factors such as time consumption, cost and individual laboratory options.

  16. Dual entanglement measures based on no local cloning and no local deleting

    International Nuclear Information System (INIS)

    Horodecki, Michal; Sen, Aditi; Sen, Ujjwal

    2004-01-01

    The impossibility of cloning and deleting of unknown states constitute important restrictions on processing of information in the quantum world. On the other hand, a known quantum state can always be cloned or deleted. However, if we restrict the class of allowed operations, there will arise restrictions on the ability of cloning and deleting machines. We have shown that cloning and deleting of known states is in general not possible by local operations. This impossibility hints at quantum correlation in the state. We propose dual measures of quantum correlation based on the dual restrictions of no local cloning and no local deleting. The measures are relative entropy distances of the desired states in a (generally impossible) perfect local cloning or local deleting process from the best approximate state that is actually obtained by imperfect local cloning or deleting machines. Just like the dual measures of entanglement cost and distillable entanglement, the proposed measures are based on important processes in quantum information. We discuss their properties. For the case of pure states, estimations of these two measures are also provided. Interestingly, the entanglement of cloning for a maximally entangled state of two two-level systems is not unity

  17. QuickStep-Cloning: a sequence-independent, ligation-free method for rapid construction of recombinant plasmids.

    Science.gov (United States)

    Jajesniak, Pawel; Wong, Tuck Seng

    2015-01-01

    Molecular cloning is an essential step in biological engineering. Methods involving megaprimer-based PCR of a whole plasmid are promising alternatives to the traditional restriction-ligation-based molecular cloning. Their widespread use, however, is hampered by some of their inherent characteristics, e.g., linear amplification, use of self-annealing megaprimers and difficulty with performing point insertion of DNA. These limitations result in low product yield and reduced flexibility in the design of a genetic construct. Here, we present a novel technique of directional cloning, which overcomes these problems yet retaining the simplicity of whole-plasmid amplification. QuickStep-Cloning utilizes asymmetric PCRs to create a megaprimer pair with 3'-overhangs, and hence, facilitates the subsequent exponential whole-plasmid amplification. QuickStep-Cloning generates nicked-circular plasmids, thereby permitting direct bacterial transformation without DNA ligation. It allows DNA fragment integration into any plasmid at any position, in an efficient, time- and cost-effective manner, without tedious intermediate DNA gel purification, modified oligonucleotides, specialty enzymes and ultra-competent cells. The method is compatible with competent E. coli cells prepared using the conventional calcium chloride method. QuickStep-Cloning expands the versatility of megaprimer-based cloning. It is an excellent addition to the cloning toolbox, for the benefit of protein engineers, metabolic engineers and synthetic biologists.

  18. Rapid CRISPR/Cas9-Mediated Cloning of Full-Length Epstein-Barr Virus Genomes from Latently Infected Cells.

    Science.gov (United States)

    Yajima, Misako; Ikuta, Kazufumi; Kanda, Teru

    2018-04-03

    Herpesviruses have relatively large DNA genomes of more than 150 kb that are difficult to clone and sequence. Bacterial artificial chromosome (BAC) cloning of herpesvirus genomes is a powerful technique that greatly facilitates whole viral genome sequencing as well as functional characterization of reconstituted viruses. We describe recently invented technologies for rapid BAC cloning of herpesvirus genomes using CRISPR/Cas9-mediated homology-directed repair. We focus on recent BAC cloning techniques of Epstein-Barr virus (EBV) genomes and discuss the possible advantages of a CRISPR/Cas9-mediated strategy comparatively with precedent EBV-BAC cloning strategies. We also describe the design decisions of this technology as well as possible pitfalls and points to be improved in the future. The obtained EBV-BAC clones are subjected to long-read sequencing analysis to determine complete EBV genome sequence including repetitive regions. Rapid cloning and sequence determination of various EBV strains will greatly contribute to the understanding of their global geographical distribution. This technology can also be used to clone disease-associated EBV strains and test the hypothesis that they have special features that distinguish them from strains that infect asymptomatically.

  19. GLUT3 is present in Clone 9 liver cells and translocates to the plasma membrane in response to insulin.

    Science.gov (United States)

    Defries, Danielle M; Taylor, Carla G; Zahradka, Peter

    2016-08-26

    Clone 9 cells have been reported to express only the GLUT1 facilitative glucose transporter; however, previous studies have not examined Clone 9 cells for GLUT3 content. The current study sought to profile the presence of glucose transporters in Clone 9 cells, H4IIE hepatoma cells, and L6 myoblasts and myotubes. While the other cell types contained the expected complement of transporters, Clone 9 cells had GLUT3 which was previously not reported. Interestingly, both GLUT3 mRNA and protein were detected in Clone 9 cells, but only mRNA for GLUT1 was detected. Glucose transport in Clone 9 cells was insulin-sensitive in a concentration-dependent manner, concomitant with the presence of GLUT3 in the plasma membrane after insulin treatment. Although basal glucose uptake was unaffected, insulin-stimulated glucose uptake was abolished with siRNA-mediated GLUT3 knockdown. These results contradict previous reports that Clone 9 cells exclusively express GLUT1 and suggest GLUT3 is a key insulin-sensitive glucose transporter required for insulin-stimulated glucose uptake by Clone 9 cells. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems.

    Science.gov (United States)

    Kirszbaum, L; Sharpe, J A; Murphy, B; d'Apice, A J; Classon, B; Hudson, P; Walker, I D

    1989-03-01

    The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems.

  1. The challenges of facilitation

    DEFF Research Database (Denmark)

    Agger, Annika

    The aim of the paper is to investigate the role of the facilitators in the enactment of the principles of deliberative democracy and how they are carried out in practice. More specifically, the focus is on how the facilitators‟ balances between the intention of opening up for a plurality of voice...

  2. Computerized adaptive testing with item clones

    NARCIS (Netherlands)

    Glas, Cornelis A.W.; van der Linden, Willem J.

    2001-01-01

    To reduce the cost of item writing and to enhance the flexibility of item presentation, items can be generated by item-cloning techniques. An important consequence of cloning is that it may cause variability on the item parameters. Therefore, a multilevel item response model is presented in which it

  3. Computerized adaptive testing with item cloning

    NARCIS (Netherlands)

    Glas, Cornelis A.W.; van der Linden, Willem J.

    2003-01-01

    To increase the number of items available for adaptive testing and reduce the cost of item writing, the use of techniques of item cloning has been proposed. An important consequence of item cloning is possible variability between the item parameters. To deal with this variability, a multilevel item

  4. The ethics of human reproductive cloning.

    Science.gov (United States)

    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.

  5. Recombination-assisted megaprimer (RAM) cloning

    Science.gov (United States)

    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

  6. Challenges in regulating farm animal cloning

    DEFF Research Database (Denmark)

    Gunning, Jennifer; Hartlev, Mette; Gamborg, Christian

    Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety......Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety...

  7. Information cloning of harmonic oscillator coherent states

    Indian Academy of Sciences (India)

    article/fulltext/pram/059/02/0263-0267. Keywords. Cloning; coherent states. Abstract. We show that in the case of unknown harmonic oscillator coherent statesit is possible to achieve what we call perfect information cloning. By this we mean that ...

  8. Reversibility of continuous-variable quantum cloning

    International Nuclear Information System (INIS)

    Filip, Radim; Marek, Petr; Fiurasek, Jaromir

    2004-01-01

    We analyze a reversibility of optimal Gaussian 1→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→M Gaussian cloning of coherent states which transforms it to optimal 1→M ' cloning for M ' < M. Assuming the quantum 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

  9. [Human cloning in Muslim and Arab law].

    Science.gov (United States)

    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.

  10. Facilitating Learning at Conferences

    DEFF Research Database (Denmark)

    Ravn, Ib; Elsborg, Steen

    2011-01-01

    The typical conference consists of a series of PowerPoint presentations that tend to render participants passive. Students of learning have long abandoned the transfer model that underlies such one-way communication. We propose an al-ternative theory of conferences that sees them as a forum...... for learning, mutual inspiration and human flourishing. We offer five design principles that specify how conferences may engage participants more and hence increase their learning. In the research-and-development effort reported here, our team collaborated with conference organizers in Denmark to introduce...... and facilitate a variety of simple learning techniques at thirty one- and two-day conferences of up to 300 participants each. We present ten of these techniques and data evaluating them. We conclude that if conference organizers allocate a fraction of the total conference time to facilitated processes...

  11. Essence: Facilitating Software Innovation

    DEFF Research Database (Denmark)

    Aaen, Ivan

    2008-01-01

      This paper suggests ways to facilitate creativity and innovation in software development. The paper applies four perspectives – Product, Project, Process, and People –to identify an outlook for software innovation. The paper then describes a new facility–Software Innovation Research Lab (SIRL) ......) – and a new method concept for software innovation – Essence – based on views, modes, and team roles. Finally, the paper reports from an early experiment using SIRL and Essence and identifies further research.......  This paper suggests ways to facilitate creativity and innovation in software development. The paper applies four perspectives – Product, Project, Process, and People –to identify an outlook for software innovation. The paper then describes a new facility–Software Innovation Research Lab (SIRL...

  12. Mindfulness for group facilitation

    DEFF Research Database (Denmark)

    Adriansen, Hanne Kirstine; Krohn, Simon

    2014-01-01

    thinking and ‘Eastern’ mindfulness which refers to an open, accepting state of mind, as intended with Buddhist-inspired techniques such as meditation. In this paper, we are interested in the latter type of mindfulness and demonstrate how Eastern mindfulness techniques can be used as a tool for facilitation......In this paper, we argue that mindfulness techniques can be used for enhancing the outcome of group performance. The word mindfulness has different connotations in the academic literature. Broadly speaking there is ‘mindfulness without meditation’ or ‘Western’ mindfulness which involves active....... A brief introduction to the physiology and philosophy of Eastern mindfulness constitutes the basis for the arguments of the effect of mindfulness techniques. The use of mindfulness techniques for group facilitation is novel as it changes the focus from individuals’ mindfulness practice...

  13. Containers, facilitators, innovators?

    DEFF Research Database (Denmark)

    Makkonen, Teemu; Merisalo, Maria; Inkinen, Tommi

    2018-01-01

    : are they containers, facilitators or innovators? This is investigated here through empirical material derived from 27 interviews with top departmental management in three Finnish cities (Helsinki, Espoo and Vantaa). The results show that local city governments (LCGs) consider cities as facilitators of innovation...... without the active role of LCGs as innovators. City employees are innovative – the seeming lack of public sector innovation is actually a result of measurement issues that favour (patentable) technological innovations rather than those more common to LCGs, meaning service and organisational types....... Therefore, LCGs can be seen as highly innovative organisations. There are, however, barriers to innovation in the public sector, such as the cost of innovation activity, the lack of incentives for it, and working culture that does not support it. Lastly, the results show that LCGs have not really fully...

  14. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach.

    Directory of Open Access Journals (Sweden)

    Hannes M Beyer

    Full Text Available 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.

  15. Facilitating Knowledge Sharing

    OpenAIRE

    Holdt Christensen, Peter

    2005-01-01

    Abstract This paper argues that knowledge sharing can be conceptualized as different situations of exchange in which individuals relate to each other in different ways, involving different rules, norms and traditions of reciprocity regulating the exchange. The main challenge for facilitating knowledge sharing is to ensure that the exchange is seen as equitable for the parties involved, and by viewing the problems of knowledge sharing as motivational problems situated in different organization...

  16. Genetic Barcodes Facilitate Competitive Clonal Analyses In Vivo.

    Science.gov (United States)

    Aranyossy, Tim; Thielecke, Lars; Glauche, Ingmar; Fehse, Boris; Cornils, Kerstin

    2017-10-01

    Monitoring the fate of individual cell clones is an important task to better understand normal tissue regeneration, for example after hematopoietic stem cell (HSC) transplantation, but also cancerogenesis. Based on their integration into the host cell's genome, retroviral vectors are commonly used to stably mark target cells and their progeny. The development of genetic barcoding techniques has opened new possibilities to determine clonal composition and dynamics in great detail. A modular genetic barcode was recently introduced consisting of 32 variable positions (BC32) with a customized backbone, and its advantages were demonstrated with regard to barcode calling and quantification. The study presented applied the BC32 system in a complex in vivo situation, namely to analyze clonal reconstitution dynamics for HSC grafts consisting of up to three cell populations with distinguishable barcodes using different alpha- and lentiviral vectors. In a competitive transplantation setup, it was possible to follow the differently marked cell populations within individual animals. This enabled the clonal contribution of the different BC32 constructs during reconstitution and long-term hematopoiesis in the peripheral blood and the spatial distribution in bone marrow and spleen to be identified. Thus, it was demonstrated that the system allows the output of individually marked cells to be tracked in vivo and their influence on clonal dynamics to be analyzed. Successful application of the BC32 system in a complex, competitive in vivo situation provided proof-of-principle that its high complexity and the large Hamming distance between individual barcodes, combined with the easy customization, facilitate efficient and precise quantification, even without prior knowledge of individual barcode sequences. Importantly, simultaneous high-sensitivity analyses of different cell populations in single animals may significantly reduce numbers of animals required to investigate specific

  17. Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning

    Directory of Open Access Journals (Sweden)

    Decai Tuo

    2015-12-01

    Full Text Available Papaya leaf distortion mosaic virus (PLDMV is becoming a threat to papaya and transgenic papaya resistant to the related pathogen, papaya ringspot virus (PRSV. The generation of infectious viral clones is an essential step for reverse-genetics studies of viral gene function and cross-protection. In this study, a sequence- and ligation-independent cloning system, the In-Fusion® Cloning Kit (Clontech, Mountain View, CA, USA, was used to construct intron-less or intron-containing full-length cDNA clones of the isolate PLDMV-DF, with the simultaneous scarless assembly of multiple viral and intron fragments into a plasmid vector in a single reaction. The intron-containing full-length cDNA clone of PLDMV-DF was stably propagated in Escherichia coli. In vitro intron-containing transcripts were processed and spliced into biologically active intron-less transcripts following mechanical inoculation and then initiated systemic infections in Carica papaya L. seedlings, which developed similar symptoms to those caused by the wild-type virus. However, no infectivity was detected when the plants were inoculated with RNA transcripts from the intron-less construct because the instability of the viral cDNA clone in bacterial cells caused a non-sense or deletion mutation of the genomic sequence of PLDMV-DF. To our knowledge, this is the first report of the construction of an infectious full-length cDNA clone of PLDMV and the splicing of intron-containing transcripts following mechanical inoculation. In-Fusion cloning shortens the construction time from months to days. Therefore, it is a faster, more flexible, and more efficient method than the traditional multistep restriction enzyme-mediated subcloning procedure.

  18. [Worldviews and philosophical basis of human cloning].

    Science.gov (United States)

    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.

  19. Human embryo cloning prohibited in Hong Kong.

    Science.gov (United States)

    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.

  20. Essence: Facilitating Software Innovation

    DEFF Research Database (Denmark)

    Aaen, Ivan

    2008-01-01

      This paper suggests ways to facilitate creativity and innovation in software development. The paper applies four perspectives – Product, Project, Process, and People –to identify an outlook for software innovation. The paper then describes a new facility–Software Innovation Research Lab (SIRL......) – and a new method concept for software innovation – Essence – based on views, modes, and team roles. Finally, the paper reports from an early experiment using SIRL and Essence and identifies further research....

  1. Feasibility of a Video-Based Advance Care Planning Website to Facilitate Group Visits among Diverse Adults from a Safety-Net Health System.

    Science.gov (United States)

    Zapata, Carly; Lum, Hillary D; Wistar, Emily; Horton, Claire; Sudore, Rebecca L

    2018-02-20

    Primary care providers in safety-net settings often do not have time to discuss advance care planning (ACP). Group visits (GV) may be an efficient means to provide ACP education. To assess the feasibility and impact of a video-based website to facilitate GVs to engage diverse adults in ACP. Feasibility pilot among patients who were ≥55 years of age from two primary care clinics in a Northern California safety-net setting. Participants attended two 90-minute GVs and viewed the five steps of the movie version of the PREPARE website ( www.prepareforyourcare.org ) concerning surrogates, values, and discussing wishes in video format. Two clinician facilitators were available to encourage participation. We assessed pre-to-post ACP knowledge, whether participants designated a surrogate or completed an advance directive (AD), and acceptability of GVs and PREPARE materials. We conducted two GVs with 22 participants. Mean age was 64 years (±7), 55% were women, 73% nonwhite, and 55% had limited literacy. Knowledge improved about surrogate designation (46% correct pre vs. 85% post, p = 0.01) and discussing decisions with others (59% vs. 90%, p = 0.01). Surrogate designation increased (48% vs. 85%, p = 0.01) and there was a trend toward AD completion (9% vs. 24%, p = 0.21). Participants rated the GVs and PREPARE materials a mean of 8 (±3.1) on a 10-point acceptability scale. Using the PREPARE movie to facilitate ACP GVs for diverse adults in safety net, primary care settings is feasible and shows potential for increasing ACP engagement.

  2. Novice facilitators and the use of scripts for managing facilitated modelling workshops

    DEFF Research Database (Denmark)

    Tavella, Elena; Papadopoulos, Thanos

    2015-01-01

    and achieve outcomes. This illustration is based on a micro-level analysis of a transcript from a Viable System Model workshop held in a food cooperative in Copenhagen, Denmark. Through our findings we identify two distinct script-supported FM behaviours and related script-supported facilitation practices......There is limited research on the use of scripts by novice facilitators (novices) in Facilitated Modelling (FM) workshops. To address this gap, this paper illustrates how novices—supported by scripts—switch between and combine facilitation skills and competencies to successfully manage FM workshops...

  3. Physiological and biochemical responses to severe drought stress of nine Eucalyptus globulus clones: a multivariate approach.

    Science.gov (United States)

    Granda, Víctor; Delatorre, Carolina; Cuesta, Candela; Centeno, María L; Fernández, Belén; Rodríguez, Ana; Feito, Isabel

    2014-07-01

    Seasonal drought, typical of temperate and Mediterranean environments, creates problems in establishing plantations and affects development and yield, and it has been widely studied in numerous species. Forestry fast-growing species such as Eucalyptus spp. are an important resource in such environments, selected clones being generally used for production purposes in plantations in these areas. However, use of mono-specific plantations increases risk of plant loss due to abiotic stresses, making it essential to understand differences in an individual clone's physiological responses to drought stress. In order to study clonal differences in drought responses, nine Eucalyptus globulus (Labill.) clones (C14, C46, C97, C120, C222, C371, C405, C491 and C601) were gradually subjected to severe drought stress (globulus clones on the basis of their different strategies when faced with drought stress. The C14 group (C14, C120, C405, C491 and C601) clones behave as water savers, maintaining high water content and showing high stomatal adjustment, and reducing their aerial growth to a great extent. The C46 group (C46, C97, C222 and C371) clones behave as water spenders, reducing their water content drastically and presenting osmotic adjustment. The latter maintains the highest growth rate under the conditions tested. The method presented here can be used to identify appropriate E. globulus clones for drought environments, facilitating the selection of material for production and repopulation environments. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Cell cloning-on-the-spot by using an attachable silicone cylinder.

    Science.gov (United States)

    Park, Hong Bum; Son, Wonseok; Chae, Dong Han; Lee, Jisu; Kim, Il-Woung; Yang, Woomi; Sung, Jae Kyu; Lim, Kyu; Lee, Jun Hee; Kim, Kyung-Hee; Park, Jong-Il

    2016-06-10

    Cell cloning is a laboratory routine to isolate and keep particular properties of cultured cells. Transfected or other genetically modified cells can be selected by the traditional microbiological cloning. In addition, common laboratory cell lines are prone to genotypic drift during their continual culture, so that supplementary cloning steps are often required to maintain correct lineage phenotypes. Here, we designed a silicone-made attachable cloning cylinder, which facilitated an easy and bona fide cloning of interested cells. This silicone cylinder was easy to make, showed competent stickiness to laboratory plastics including culture dishes, and hence enabled secure isolation and culture for days of selected single cells, especially, on the spots of preceding cell-plating dishes under microscopic examination of visible cellular phenotypes. We tested the silicone cylinder in the monoclonal subcloning from a heterogeneous population of a breast cancer cell line, MDA-MB-231, and readily established independent MDA-MB-231 subclones showing different sublineage phenotypes. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Cloning and functional expression of a human pancreatic islet glucose-transporter cDNA

    International Nuclear Information System (INIS)

    Permutt, M.A.; Koranyi, L.; Keller, K.; Lacy, P.E.; Scharp, D.W.; Mueckler, M.

    1989-01-01

    Previous studies have suggested that pancreatic islet glucose transport is mediated by a high-K m , low-affinity facilitated transporter similar to that expressed in liver. To determine the relationship between islet and liver glucose transporters, liver-type glucose-transporter cDNA clones were isolated from a human liver cDNA library. The liver-type glucose-transporter cDNA clone hybridized to mRNA transcripts of the same size in human liver and pancreatic islet RNA. A cDNA library was prepared from purified human pancreatic islet tissue and screened with human liver-type glucose-transporter cDNA. The authors isolated two overlapping cDNA clones encompassing 2600 base pairs, which encode a pancreatic islet protein identical in sequence to that of the putative liver-type glucose-transporter protein. Xenopus oocytes injected with synthetic mRNA transcribed from a full-length cDNA construct exhibited increased uptake of 2-deoxyglucose, confirming the functional identity of the clone. These cDNA clones can now be used to study regulation of expression of the gene and to assess the role of inherited defects in this gene as a candidate for inherited susceptibility to non-insulin-dependent diabetes mellitus

  6. Cloning and Penal Limitations of Scientific Experiment

    Directory of Open Access Journals (Sweden)

    Rustin-Petru CIASC

    2014-03-01

    Full Text Available Human cloning is one of the matters that have been broadly discussed at a scientific and legal level. In this article, I will present the worldwide relevant aspects as regards this. The bioethical substantiation of forbidding human cloning is made, first, by the fact that the artificial cloning of a human being is a threat to the human identity because it endangers the protection against predetermining the human genetic constitution by a third party; the human dignity is thus endangered by transforming the human being into an object by artificial cloning. Given the insufficient coverage that the national legislation provides to this topic, according to the legislative experience belonging to other countries, this article presents some legislative proposals in order to fill in at least partially such gaps.

  7. DNA microarrays : a molecular cloning manual

    National Research Council Canada - National Science Library

    Sambrook, Joseph; Bowtell, David

    2002-01-01

    .... This manual, designed to extend and to complement the information in the best-selling Molecular Cloning, is a synthesis of the expertise and experience of more than 30 contributors all innovators in a fast moving field...

  8. Generation of phase-covariant quantum cloning

    International Nuclear Information System (INIS)

    Karimipour, V.; Rezakhani, A.T.

    2002-01-01

    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

  9. A new β-estradiol-inducible vector set that facilitates easy construction and efficient expression of transgenes reveals CBL3-dependent cytoplasm to tonoplast translocation of CIPK5.

    Science.gov (United States)

    Schlücking, Kathrin; Edel, Kai H; Köster, Philipp; Drerup, Maria M; Eckert, Christian; Steinhorst, Leonie; Waadt, Rainer; Batistic, Oliver; Kudla, Jörg

    2013-11-01

    Transient and stable expression of transgenes is central to many investigations in plant biology research. Chemical regulation of expression can circumvent problems of plant lethality caused by constitutive overexpression or allow inducible knock (out/down) approaches. Several chemically inducible or repressible systems have been described and successfully applied. However, cloning and application-specific modification of most available inducible expression systems have been limited and remained complicated due to restricted cloning options. Here we describe a new set of 57 vectors that enable transgene expression in transiently or stably transformed cells. All vectors harbor a synthetically optimized XVE expression cassette, allowing β-estradiol mediated protein expression. Plasmids are equipped with the reporter genes GUS, GFP, mCherry, or with HA and StrepII epitope tags and harbor an optimized multiple cloning site for flexible and simple cloning strategies. Moreover, the vector design allows simple substitution of the driving promoter to achieve tissue-specificity or to modulate expression ranges of inducible transgene expression. We report details of the kinetics and dose-dependence of expression induction in Arabidopsis leaf mesophyll protoplasts, transiently transformed Nicotiana benthamiana leaves, and stably transformed Arabidopsis plants. Using these vectors, we investigated the influence of CBL (Calcineurin B-like) protein expression on the subcellular localization of CIPKs (Calcineurin B-like interacting protein kinases). These analyses uncovered that induced co-expression of CBL3 is fully sufficient for dynamic translocation of CIPK5 from the cytoplasm to the tonoplast. Thus, the vector system presented here facilitates a broad range of research applications.

  10. Speech acts and performances of scientific citizenship: Examining how scientists talk about therapeutic cloning.

    Science.gov (United States)

    Marks, Nicola J

    2014-07-01

    Scientists play an important role in framing public engagement with science. Their language can facilitate or impede particular interactions taking place with particular citizens: scientists' "speech acts" can "perform" different types of "scientific citizenship". This paper examines how scientists in Australia talked about therapeutic cloning during interviews and during the 2006 parliamentary debates on stem cell research. Some avoided complex labels, thereby facilitating public examination of this field. Others drew on language that only opens a space for publics to become educated, not to participate in a more meaningful way. Importantly, public utterances made by scientists here contrast with common international utterances: they did not focus on the therapeutic but the research promises of therapeutic cloning. Social scientists need to pay attention to the performative aspects of language in order to promote genuine citizen involvement in techno-science. Speech Act Theory is a useful analytical tool for this.

  11. Cloning and Biochemical Characterization of HIV Integrase

    Science.gov (United States)

    1989-12-15

    biochemical assays for the interaction of the HIV Intebration protein, IN, with a specific DNA target, the viral LTRs. The gene encoding IN has been subcloned ...FILE COPY CO CONTRACT NO.: DAMD7-88-C-8126 r TITLE: CLONING AND BIOCHEMICAL CHARACTERIZATION OF HIV INTEGRASE I- SPRINCIPAL INVESTIGATOR: ELLEN...ELEMENT No. NO. 392- NO. IACCESSION NO. 1 . TITLE (indude Security Classification)630A I315D49C08 (U) Cloning and Biochemical Characterization of the HIV

  12. Endangered wolves cloned from adult somatic cells.

    Science.gov (United States)

    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.

  13. Molecular cloning of feline immunodeficiency virus.

    OpenAIRE

    Olmsted, R A; Barnes, A K; Yamamoto, J K; Hirsch, V M; Purcell, R H; Johnson, P R

    1989-01-01

    Feline immunodeficiency virus (FIV) is a T-lymphotropic retrovirus associated with immunodeficiency and opportunistic infections in cats. The discovery of FIV provides an opportunity for the development of a small animal model for AIDS. To initiate the molecular and biological characterization of FIV, cDNA clones were synthesized and used to isolate a proviral clone of FIV. Molecular cross-hybridization analysis of FIV with five lentiviruses revealed that nucleotide-sequence similarities exis...

  14. Birth of viable puppies derived from breeding cloned female dogs with a cloned male.

    Science.gov (United States)

    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.

  15. Do Clones Dream of Love? Images of Clones in Popular Culture

    Directory of Open Access Journals (Sweden)

    Dragana Antonijević

    2016-02-01

    Full Text Available Fantasies about clones, cyborgs and androids have become part and parcel of the mythology of modern times – the mythologies of the biotechnological era in which the achievements of genetic engineering have inflamed fears of possible abuse of scientific knowledge and the consequences of such abuse. The paper considers the phenomenon of reproductive cloning of human beings as it is represented in popular culture, especially film as it is one of the most important sources of representations and constructions of ideas about clones. After the introductory consideration of this phenomenon in scientific, ethical and media debates which are imbued with rejection of reproductive cloning, I have analyzed the different uses of the clone motif in selected movies. I have examined the structure and content of the genre formula of "social melodrama" which is present in films about clones, and have analyzed the mythical patterns pertaining to the topic of cloning, such as the myth of immortality, the myth of twins, the myth of the uniqueness of human kind etc. Ultimately, the nature and origins of the fear of clones and disgust that clones cause have been examined, and it has been shown that they mostly boil down to the fear of the dehumanization of human beings, the fear of the loss of difference and the transgression of biological, sociocultural and metaphysical boundaries.

  16. [CDNA cloning of human leptin and its expression].

    Science.gov (United States)

    Jia, Zhen-Yu; Fu, Xiao-Min; Jin, Ai-Hua; Cao, Jiang

    2003-07-01

    To clone cDNA of human leptin gene and obtain leptin protein for future study on leptin binding proteins. The cDNA of human leptin with 6 x his-tag was cloned by over-hang extension PCR protocol using human genomic DNA as template, and subcloned into in vitro expression vector pIVEX2.3MCS, and the fusion protein was expressed in vitro by Rapid Translation System (RTS) (RTS500 cycle primer Kit and RTS500 ProteoMaster of Roche company). The apparent molecular weight(19.46 kD) and the immuno-specificity of the fusion protein were confirmed by SDS-PAGE and Western blot, and the expressed fusion protein stayed mainly in the supernatant of the reaction mixture in soluble form. This work provides us solid basis for further study on new leptin-associated proteins.

  17. Emotional reactions to human reproductive cloning.

    Science.gov (United States)

    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/

  18. Cloning: Past, Present, and the Exciting Future. Breakthroughs in Bioscience.

    Science.gov (United States)

    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)

  19. Public perceptions of farm animal cloning in Europe

    DEFF Research Database (Denmark)

    Lassen, Jesper

    This report presents a picture of European opinion on farm animal cloning. In the report, both agricultural and biomedical applications of farm animal cloning are considered. With the arrival of Dolly, animal cloning became an integral part of the biotech debate, but this debate did not isolate...... animal cloning as a single issue....

  20. Cloning the entanglement of a pair of quantum bits

    International Nuclear Information System (INIS)

    Lamoureux, Louis-Philippe; Navez, Patrick; Cerf, Nicolas J.; Fiurasek, Jaromir

    2004-01-01

    It is shown that any quantum operation that perfectly clones the entanglement of all maximally entangled qubit pairs cannot preserve separability. This 'entanglement no-cloning' principle naturally suggests that some approximate cloning of entanglement is nevertheless allowed by quantum mechanics. We investigate a separability-preserving optimal cloning machine that duplicates all maximally entangled states of two qubits, resulting in 0.285 bits of entanglement per clone, while a local cloning machine only yields 0.060 bits of entanglement per clone

  1. Clone and characterization of photolyase-gene from soybean

    International Nuclear Information System (INIS)

    Najrana, T.; Hirouchi, T.; Yamamoto, K.

    2003-01-01

    Full text: Cyclobutane pyrimidine dimer (CPD) and pyrimidine [6-4] pyrimidone photoproduct (6-4pp) are the major products of UV-radiation. Both CPD and 6-4pp posses lethal as well as mutagenic property. Excision repair and photoreactivation are involved as major pathways in repairing those photoproducts. To repair those products plant uses photoreactivation as a major pathway. In photoreactivation process photolyase (enzyme encoded by PHR-gene) catalyzes the splitting of the dimer into a monomer under blue light. Photolyase is specific for damage CPD or 6-4pp. The CPD and 6-4pp photolyases are responsible for repairing CPD and 6-4pp lesions respectively. Several investigators reported that removal of CPD lesion is necessary for survival in higher plants in the early development. Thus one should realize the importance of clone and characterization of CPD-photolyase gene from plants especially from those are lying in the list of foods such as wheat, corn, soybean etc. cDNA library (pSPORT-P) of soybean was amplified using the primers that designated as common for CPD-photolyase gene for plants. These primers gave the desire size of PCR product. Desirable PCR product inserted into TA-cloning vector and sequenced. Amino acid sequence revealed considerable homology with CPD-photolyases of rice, arabidopsis thaliana. Then using dilution-PCR amplification method (Hirouchi et al., MGG in press) I have identified the true clone from cDNA library of soybean that containing the full length of CPD-photolyase gene. Full length of cloned gene is about 1698 bps long and exist start and stop codon. Amino acid sequence of the cloned gene shows more than 70% homology with rice, arabidopsis thaliana. Cloned gene enables to complement the E. coli ( phr-uvrA-recA-) system that is completely defective in photoreactivation. The size of CPD-photolyase of soybean is about 56 KDa as identified by 12% SDS PAGE

  2. A web-based system to facilitate local, systematic quality improvement by multidisciplinary care teams: development and first experiences of CARDSS Online

    NARCIS (Netherlands)

    van Engen-Verheul, Mariëtte M.; van der Veer, Sabine N.; de Keizer, Nicolette F.; Tjon Sjoe Sjoe, Winston; van der Zwan, Eric P. A.; Peek, Niels

    2013-01-01

    Continuous monitoring and systematic improvement of quality have become increasingly common in healthcare. To support multidisciplinary care teams in improving their clinical performance using feedback on quality indicators, we developed the CARDSS Online system. This system supports (i) monitoring

  3. Facilitative root interactions in intercrops

    DEFF Research Database (Denmark)

    Hauggaard-Nielsen, H.; Jensen, E.S.

    2005-01-01

    Facilitation takes place when plants ameliorate the environment of their neighbours, and increase their growth and survival. Facilitation occurs in natural ecosystems as well as in agroecosystems. We discuss examples of facilitative root interactions in intercropped agroecosystems; including...... of root architecture, exudation of growth stimulating substances, and biofumigation. Facilitative root interactions are most likely to be of importance in nutrient poor soils and in low-input agroecosystems due to critical interspecific competition for plant growth factors. However, studies from more...

  4. Does cerebral hypoxia facilitate central fatigue?

    DEFF Research Database (Denmark)

    Siebenmann, Christoph; Rasmussen, Peter

    2016-01-01

    Central fatigue originates within the central nervous system and is characterized by a decreasein voluntary muscle activation. Reduced systemic O2availability can facilitate central fatigue byenhancing the afferent input of the chemosensitive nerves that play a pivotal role in developmentof centr...

  5. Facilitating Learning Organizations. Making Learning Count.

    Science.gov (United States)

    Marsick, Victoria J.; Watkins, Karen E.

    This book offers advice to facilitators and change agents who wish to build systems-level learning to create knowledge that can be used to gain a competitive advantage. Chapter 1 describes forces driving companies to build, sustain, and effectively use systems-level learning and presents and links a working definition of the learning organization…

  6. Synthesis and cell-free cloning of DNA libraries using programmable microfluidics.

    Science.gov (United States)

    Ben Yehezkel, Tuval; Rival, Arnaud; Raz, Ofir; Cohen, Rafael; Marx, Zipora; Camara, Miguel; Dubern, Jean-Frédéric; Koch, Birgit; Heeb, Stephan; Krasnogor, Natalio; Delattre, Cyril; Shapiro, Ehud

    2016-02-29

    Microfluidics may revolutionize our ability to write synthetic DNA by addressing several fundamental limitations associated with generating novel genetic constructs. Here we report the first de novo synthesis and cell-free cloning of custom DNA libraries in sub-microliter reaction droplets using programmable digital microfluidics. Specifically, we developed Programmable Order Polymerization (POP), Microfluidic Combinatorial Assembly of DNA (M-CAD) and Microfluidic In-vitro Cloning (MIC) and applied them to de novo synthesis, combinatorial assembly and cell-free cloning of genes, respectively. Proof-of-concept for these methods was demonstrated by programming an autonomous microfluidic system to construct and clone libraries of yeast ribosome binding sites and bacterial Azurine, which were then retrieved in individual droplets and validated. The ability to rapidly and robustly generate designer DNA molecules in an autonomous manner should have wide application in biological research and development. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  7. Connecting Smartphone and Wearable Fitness Tracker Data with a Nationally Used Electronic Health Record System for Diabetes Education to Facilitate Behavioral Goal Monitoring in Diabetes Care: Protocol for a Pragmatic Multi-Site Randomized Trial.

    Science.gov (United States)

    Wang, Jing; Coleman, Deidra Carroll; Kanter, Justin; Ummer, Brad; Siminerio, Linda

    2018-04-02

    Mobile and wearable technology have been shown to be effective in improving diabetes self-management; however, integrating data from these technologies into clinical diabetes care to facilitate behavioral goal monitoring has not been explored. The objective of this paper is to report on a study protocol for a pragmatic multi-site trial along with the intervention components, including the detailed connected health interface. This interface was developed to integrate patient self-monitoring data collected from a wearable fitness tracker and its companion smartphone app to an electronic health record system for diabetes self-management education and support (DSMES) to facilitate behavioral goal monitoring. A 3-month multi-site pragmatic clinical trial was conducted with eligible patients with diabetes mellitus from DSMES programs. The Chronicle Diabetes system is currently freely available to diabetes educators through American Diabetes Association-recognized DSMES programs to set patient nutrition and physical activity goals. To integrate the goal-setting and self-monitoring intervention into the DSMES process, a connected interface in the Chronicle Diabetes system was developed. With the connected interface, patient self-monitoring information collected from smartphones and wearable fitness trackers can facilitate educators' monitoring of patients' adherence to their goals. Feasibility outcomes of the 3-month trial included hemoglobin A 1c levels, weight, and the usability of the connected system. An interface designed to connect data from a wearable fitness tracker with a companion smartphone app for nutrition and physical activity self-monitoring into a diabetes education electronic health record system was successfully developed to enable diabetes educators to facilitate goal setting and monitoring. A total of 60 eligible patients with type 2 diabetes mellitus were randomized into either group 1) standard diabetes education or 2) standard education enhanced with

  8. Whole genome comparison of donor and cloned dogs

    OpenAIRE

    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 ...

  9. Customer and Developers Point of Views to Game Cloning

    Directory of Open Access Journals (Sweden)

    Ryan Susanto

    2014-11-01

    Full Text Available Game Cloning is an action of cloning games. This non-ethical action reaps many controversial matters from game customer and developer in game industry. As a result, this action is considered a wrongdoing. However, but not all cases of game cloning are wrong. Many recent games have been rooted from game cloning. In case of game cloning, game developers do not commit something illegal or immoral since they revamp and complement the games they imitate

  10. HOW TO FACILITATE THE MOVEMENT OF PASSENGERS BY INTRODUCING BAGGAGE COLLECTION SYSTEMS FOR TRAVEL FROM NORTH SHIELDS TO NEWCASTLE INTERNATIONAL AIRPORT

    Directory of Open Access Journals (Sweden)

    Daniel Connor REECE

    2015-12-01

    Full Text Available This paper reviews current systems that either transport baggage or have the potential to transport baggage, as well as proposed systems, before modelling some of them in a modal choice model to examine how effective they would be if implemented. It was hoped that the introduction of a baggage collection system would increase the number of passengers using public transport which would be good for the environment. The specific location of the study was Tyne and Wear and a journey from North Shields to Newcastle International Airport was chosen for the analysis. The system proposed is compared to an existing systems known as InPost and Virgin Bag Magic. It was found that for the average passenger in North Shields, a system based on the way InPost operates would offer the greatest utility. However, with the introduction of a new utility equation that could analyse mixed-mode travel, a baggage collection hub based in Newcastle upon Tyne city centre offered a more significant number of users provided that the cost of the system was either covered in the travel ticket or provided by the airport or airline free of charge. This dedicated baggage collection system would be much more expensive to introduce compared to the InPost system as the infrastructure to run the InPost system is already in place, however, the ridership of the newly proposed system would be much larger therefore it could potentially recoup the development costs.

  11. Ribosomal binding site switching: an effective strategy for high-throughput cloning constructions.

    Directory of Open Access Journals (Sweden)

    Yangbo Hu

    Full Text Available Direct cloning of PCR fragments by TA cloning or blunt end ligation are two simple methods which would greatly benefit high-throughput (HTP cloning constructions if the efficiency can be improved. In this study, we have developed a ribosomal binding site (RBS switching strategy for direct cloning of PCR fragments. RBS is an A/G rich region upstream of the translational start codon and is essential for gene expression. Change from A/G to T/C in the RBS blocks its activity and thereby abolishes gene expression. Based on this property, we introduced an inactive RBS upstream of a selectable marker gene, and designed a fragment insertion site within this inactive RBS. Forward and reverse insertions of specifically tailed fragments will respectively form an active and inactive RBS, thus all background from vector self-ligation and fragment reverse insertions will be eliminated due to the non-expression of the marker gene. The effectiveness of our strategy for TA cloning and blunt end ligation are confirmed. Application of this strategy to gene over-expression, a bacterial two-hybrid system, a bacterial one-hybrid system, and promoter bank construction are also verified. The advantages of this simple procedure, together with its low cost and high efficiency, makes our strategy extremely useful in HTP cloning constructions.

  12. SSRI Facilitated Crack Dancing

    Directory of Open Access Journals (Sweden)

    Ravi Doobay

    2017-01-01

    Full Text Available Choreoathetoid movement secondary to cocaine use is a well-documented phenomenon better known as “crack dancing.” It consists of uncontrolled writhing movements secondary to excess dopamine from cocaine use. We present a 32-year-old male who had been using cocaine for many years and was recently started on paroxetine, a selective serotonin reuptake inhibitor (SSRI for worsening depression four weeks before presentation. He had been doing cocaine every 2 weeks for the last three years and had never “crack danced” before this episode. The authors have conducted a thorough literature review and cited studies that suggest “crack dancing” is associated with excess dopamine. There has never been a documented case report of an SSRI being linked with “crack dancing.” The authors propose that the excess dopaminergic effect of the SSRI lowered the dopamine threshold for “crack dancing.” There is a communication with the Raphe Nucleus and the Substantia Nigra, which explains how the SSRI increases dopamine levels. This is the first documented case of an SSRI facilitating the “crack dance.”

  13. Facilitating post traumatic growth

    Directory of Open Access Journals (Sweden)

    Cox Helen

    2004-07-01

    Full Text Available Abstract Background Whilst negative responses to traumatic injury have been well documented in the literature, there is a small but growing body of work that identifies posttraumatic growth as a salient feature of this experience. We contribute to this discourse by reporting on the experiences of 13 individuals who were traumatically injured, had undergone extensive rehabilitation and were discharged from formal care. All participants were injured through involvement in a motor vehicle accident, with the exception of one, who was injured through falling off the roof of a house. Methods In this qualitative study, we used an audio-taped in-depth interview with each participant as the means of data collection. Interviews were transcribed verbatim and analysed thematically to determine the participants' unique perspectives on the experience of recovery from traumatic injury. In reporting the findings, all participants' were given a pseudonym to assure their anonymity. Results Most participants indicated that their involvement in a traumatic occurrence was a springboard for growth that enabled them to develop new perspectives on life and living. Conclusion There are a number of contributions that health providers may make to the recovery of individuals who have been traumatically injured to assist them to develop new views of vulnerability and strength, make changes in relationships, and facilitate philosophical, physical and spiritual growth.

  14. Expansion of the gateway multisite recombination cloning toolkit.

    Science.gov (United States)

    Shearin, Harold K; Dvarishkis, Alisa R; Kozeluh, Craig D; Stowers, R Steven

    2013-01-01

    Precise manipulation of transgene expression in genetic model organisms has led to advances in understanding fundamental mechanisms of development, physiology, and genetic disease. Transgene construction is, however, a precondition of transgene expression, and often limits the rate of experimental progress. Here we report an expansion of the modular Gateway MultiSite recombination-cloning platform for high efficiency transgene assembly. The expansion includes two additional destination vectors and entry clones for the LexA binary transcription system, among others. These new tools enhance the expression levels possible with Gateway MultiSite generated transgenes and make possible the generation of LexA drivers and reporters with Gateway MultiSite cloning. In vivo data from transgenic Drosophila functionally validating each novel component are presented and include neuronal LexA drivers, LexAop2 red and green fluorescent synaptic vesicle reporters, TDC2 and TRH LexA, GAL4, and QF drivers, and LexAop2, UAS, and QUAS channelrhodopsin2 T159C reporters.

  15. Facilitating attachment after international adoption.

    Science.gov (United States)

    Murphy, Natalie L

    2009-01-01

    Americans have increasingly turned to international adoption (IA) as an alternative way to build a family. Unfortunately, IA families are often being developed under conditions of loss, and sometimes these families struggle to form healthy attachments to each other. Disordered attachment (the failure to form a reciprocal, loving bond between parent and child) can occur, and can have devastating consequences. In some instances, IA children have been relinquished into state foster care systems; other families simply struggle for years caring for a developmentally delayed child who appears to have no emotion for his/her adoptive family. Nurses are likely to have contact with IA families and can use their education about attachment and bonding to help facilitate attachment in these developing families. Swanson's caring theory provides a clinically useful guide to meet this need.

  16. Cloning mice and men: prohibiting the use of iPS cells for human reproductive cloning.

    Science.gov (United States)

    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.

  17. Update on the First Cloned Dog and Outlook for Canine Cloning.

    Science.gov (United States)

    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.

  18. The type three secretion system facilitates migration of Burkholderia terrae BS001 in the mycosphere of two soil-borne fungi

    NARCIS (Netherlands)

    Yang, Pu; Zhang, Miaozhi; Warmink, Jan Aaldrik; Wang, Miao; van Elsas, Jan Dirk

    2016-01-01

    The type three secretion system (T3SS) is known to play a critical role in several bacterial-eukaryotic cell interactions. Recent indirect evidence has also pointed to a role of this system in bacterial-fungal interactions in soil. In the current study, we examine if the T3SS of the

  19. Elephant grass clones for silage production

    Directory of Open Access Journals (Sweden)

    Rerisson José Cipriano dos Santos

    2013-02-01

    Full Text Available Ensiling warm-season grasses often requires wilting due to their high moisture content, and the presence of low-soluble sugars in these grasses usually demands the use of additives during the ensiling process. This study evaluated the bromatological composition of the fodder and silage from five Pennisetum sp. clones (IPA HV 241, IPA/UFRPE Taiwan A-146 2.114, IPA/UFRPE Taiwan A-146 2.37, Elephant B, and Mott. The contents of 20 Polyvinyl chloride (PVC silos, which were opened after 90 days of storage, were used for the bromatological analysis and the evaluation of the pH, nitrogen, ammonia, buffer capacity, soluble carbohydrates, and fermentation coefficients. The effluent losses, gases and dry matter recovery were also calculated. Although differences were observed among the clones (p < 0.05 for the concentrations of dry matter, insoluble nitrogen in acid detergents, insoluble nitrogen in neutral detergents, soluble carbohydrates, fermentation coefficients, and in vitro digestibility in the forage before ensiling, no differences were observed for most of these variables after ensiling. All of the clones were efficient in the fermentation process. The IPA/UFRPE TAIWAN A-146 2.37 clone, however, presented a higher dry matter concentration and the best fermentation coefficient, resulting in a better silage quality, compared to the other clones.

  20. Nano-sized Fe2O3/Fe3O4 facilitate anaerobic transformation of hexavalent chromium in soil-water systems.

    Science.gov (United States)

    Zhang, Yaxian; Li, Hua; Gong, Libo; Dong, Guowen; Shen, Liang; Wang, Yuanpeng; Li, Qingbiao

    2017-07-01

    The purpose of this study is to investigate the effects of nano-sized or submicro Fe 2 O 3 /Fe 3 O 4 on the bioreduction of hexavalent chromium (Cr(VI)) and to evaluate the effects of nano-sized Fe 2 O 3 /Fe 3 O 4 on the microbial communities from the anaerobic flooding soil. The results indicated that the net decreases upon Cr(VI) concentration from biotic soil samples amended with nano-sized Fe 2 O 3 (317.1±2.1mg/L) and Fe 3 O 4 (324.0±22.2mg/L) within 21days, which were approximately 2-fold of Cr(VI) concentration released from blank control assays (117.1±5.6mg/L). Furthermore, the results of denaturing gradient gel electrophoresis (DGGE) and high-throughput sequencing indicated a greater variety of microbes within the microbial community in amendments with nano-sized Fe 2 O 3 /Fe 3 O 4 than the control assays. Especially, Proteobacteria occupied a predominant status on the phylum level within the indigenous microbial communities from chromium-contaminated soils. Besides, some partial decrease of soluble Cr(VI) in abiotic nano-sized Fe 2 O 3 /Fe 3 O 4 amendments was responsible for the adsorption of nano-sized Fe 2 O 3 /Fe 3 O 4 to soluble Cr(VI). Hence, the presence of nano-sized Fe 2 O 3 /Fe 3 O 4 could largely facilitate the mobilization and biotransformation of Cr(VI) from flooding soils by adsorption and bio-mediated processes. Copyright © 2017. Published by Elsevier B.V.

  1. Technology-facilitated depression care management among predominantly Latino diabetes patients within a public safety net care system: comparative effectiveness trial design.

    Science.gov (United States)

    Wu, Shinyi; Ell, Kathleen; Gross-Schulman, Sandra G; Sklaroff, Laura Myerchin; Katon, Wayne J; Nezu, Art M; Lee, Pey-Jiuan; Vidyanti, Irene; Chou, Chih-Ping; Guterman, Jeffrey J

    2014-03-01

    Health disparities in minority populations are well recognized. Hispanics and Latinos constitute the largest ethnic minority group in the United States; a significant proportion receives their care via a safety net. The prevalence of diabetes mellitus and comorbid depression is high among this group, but the uptake of evidence-based collaborative depression care management has been suboptimal. The study design and baseline characteristics of the enrolled sample in the Diabetes-Depression Care-management Adoption Trial (DCAT) establishes a quasi-experimental comparative effectiveness research clinical trial aimed at accelerating the adoption of collaborative depression care in safety net clinics. The study was conducted in collaboration with the Los Angeles County Department of Health Services at eight county-operated clinics. DCAT has enrolled 1406 low-income, predominantly Hispanic/Latino patients with diabetes to test a translational model of depression care management. This three-group study compares usual care with a collaborative care team support model and a technology-facilitated depression care model that provides automated telephonic depression screening and monitoring tailored to patient conditions and preferences. Call results are integrated into a diabetes disease management registry that delivers provider notifications, generates tasks, and issues critical alerts. All subjects receive comprehensive assessments at baseline, 6, 12, and 18 months by independent English-Spanish bilingual interviewers. Study outcomes include depression outcomes, treatment adherence, satisfaction, acceptance of assessment and monitoring technology, social and economic stress reduction, diabetes self-care management, health care utilization, and care management model cost and cost-effectiveness comparisons. DCAT's goal is to optimize depression screening, treatment, follow-up, outcomes, and cost savings to reduce health disparities. Copyright © 2013 Elsevier Inc. All rights

  2. Molecular cloning and evolutionary analysis of the GJA1 (connexin43) gene from bats (Chiroptera).

    Science.gov (United States)

    Wang, Li; Li, Gang; Wang, Jinhong; Ye, Shaohui; Jones, Gareth; Zhang, Shuyi

    2009-04-01

    Gap junction protein connexin43 (Cx43), encoded by the GJA1 gene, is the most abundant connexin in the cardiovascular system and was reported as a crucial factor maintaining cardiac electrical conduction, as well as having a very important function in facilitating the recycling of potassium ions from hair cells in the cochlea back into the cochlear endolymph during auditory transduction processes. In mammals, bats are the only taxon possessing powered flight, placing exceptional demand on many organismal processes. To meet the demands of flying, the hearts of bats show many specialties. Moreover, ultrasonic echolocation allows bat species to orientate and often detect and locate food in darkness. In this study, we cloned the full-length coding region of GJA1 gene from 12 different species of bats and obtained orthologous sequences from other mammals. We used the maximum likelihood method to analyse the evolution of GJA1 gene in mammals and the lineage of bats. Our results showed this gene is much conserved in mammals, as well as in bats' lineage. Compared with other mammals, we found one private amino acid substitution shared by bats, which is located on the inner loop domain, as well as some species-specific amino acid substitutions. The evolution rate analyses showed the signature of purifying selection on not only different classification level lineages but also the different domains and amino acid residue sites of this gene. Also, we suggested that GJA1 gene could be used as a good molecular marker to do the phylogenetic reconstruction.

  3. Molecular cloning and characterization of a new peptide deformylase from human pathogenic bacterium Helicobacter pylori

    International Nuclear Information System (INIS)

    Han Cong; Wang Qi; Dong Lei; Sun Haifang; Peng Shuying; Chen Jing; Yang Yiming; Yue Jianmin; Shen Xu; Jiang Hualiang

    2004-01-01

    Helicobacter pylori is a gram-negative pathogenic bacterium, which is associated with peptic ulcer disease and gastric cancer. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. In bacteria, peptide deformylase (PDF) catalyzes the removal of a formyl group from the N-termini of nascent polypeptides. Due to its essentiality and absence in mammalian cells, PDF has been considered as an attractive target for the discovery of novel antibiotics. In this work, a new PDF gene (def) from H. pylori strain SS1 was cloned, expressed, and purified in Escherichia coli system. Sequence alignment shows that H. pylori PDF (HpPDF) shares about 40% identity to E. coli PDF (EcPDF). The enzymatic properties of HpPDF demonstrate its relatively high activity toward formyl-Met-Ala-Ser, with K cat of 3.4 s -1 , K m of 1.7 mM, and K cat /K m of 2000 M -1 s -1 . HpPDF enzyme appears to be fully active at pH between 8.0 and 9.0, and temperature 50 deg. C. The enzyme activity of Co 2+ -containing HpPDF is apparently higher than that of Zn 2+ -containing HpPDF. This present work thereby supplies a potential platform that facilitates the discovery of novel HpPDF inhibitors and further of possible antimicrobial agents against H. pylori

  4. Molecular cloning and characterization of a new peptide deformylase from human pathogenic bacterium Helicobacter pylori.

    Science.gov (United States)

    Han, Cong; Wang, Qi; Dong, Lei; Sun, Haifang; Peng, Shuying; Chen, Jing; Yang, Yiming; Yue, Jianmin; Shen, Xu; Jiang, Hualiang

    2004-07-09

    Helicobacter pylori is a gram-negative pathogenic bacterium, which is associated with peptic ulcer disease and gastric cancer. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. In bacteria, peptide deformylase (PDF) catalyzes the removal of a formyl group from the N-termini of nascent polypeptides. Due to its essentiality and absence in mammalian cells, PDF has been considered as an attractive target for the discovery of novel antibiotics. In this work, a new PDF gene (def) from H. pylori strain SS1 was cloned, expressed, and purified in Escherichia coli system. Sequence alignment shows that H. pylori PDF (HpPDF) shares about 40% identity to E. coli PDF (EcPDF). The enzymatic properties of HpPDF demonstrate its relatively high activity toward formyl-Met-Ala-Ser, with K(cat) of 3.4s(-1), K(m) of 1.7 mM, and K(cat) / K(m) of 2000M(-1)s(-1). HpPDF enzyme appears to be fully active at pH between 8.0 and 9.0, and temperature 50 degrees C. The enzyme activity of Co(2+)-containing HpPDF is apparently higher than that of Zn(2+)-containing HpPDF. This present work thereby supplies a potential platform that facilitates the discovery of novel HpPDF inhibitors and further of possible antimicrobial agents against H. pylori.

  5. In vitro differentiation from a pluripotent human CD4+CD8+ thymic cloned cell into four phenotypically distinct subsets.

    Science.gov (United States)

    Boumsell, L; Schmid, M; Dastot, H; Gouttefangeas, C; Mathieu-Mahul, D; Bensussan, A

    1990-11-01

    Human thymic cell differentiation is almost totally unknown. In the present study we developed an in vitro system using human thymic cloned cells to analyze precursor-progeny relationship. We obtained several CD4+CD8+ double positive thymic clones that could give rise after several weeks in culture only to either CD4 or CD8 single positive clones. By contrast we isolated a unique pluripotent thymic double positive clone, termed B12, which differentiated into four phenotypically distinct T cell clones, namely double-positive CD4+CD8+, double-negative CD4-CD8- or either single-positive phenotype. We derived stable subclones representative of each phenotype and we showed by molecular analysis that they expressed the same TCR. Utilization of either CD3 or anticlonotypic mAb revealed that this TCR expressed by the four subclones was functional.

  6. Overexpressing tagged proteins in plants using a modified gateway cloning strategy.

    Science.gov (United States)

    Dubin, Manu J; Bowler, Chris; Benvenuto, Giovanna

    2010-03-01

    In recent years, sequence-specific recombination cloning methods such as the Gateway system have become increasingly popular for (over)expressing tagged proteins in high-throughput investigations in many different organisms, including plants. Because of their versatility and ease of use, these methods have gained favor in low- and medium-throughput investigations as well. However, due to the recombination step, the resulting fusion proteins contain long and often highly charged polylinker sequences that can interfere with their physiological function. Furthermore, in some cases the gene of interest must be cloned twice (once with and once without a stop codon) for N- and C-terminal tagging. Here, we present a hybrid combinatorial cloning strategy that overcomes many of these limitations. In the first step, the gene of interest is cloned into an entry vector containing standardized cloning sites with the desired N- or C-terminal tag and an optimized polylinker sequence. A Gateway recombination reaction is used to transfer the protein-tag fusion from the entry clone to a Gateway destination vector with the desired promoter and selectable marker for the organism of interest. As experimental requirements evolve, constructs for expressing the protein of interest with the desired tag, promoter, and selectable marker or other features can rapidly and easily be created.

  7. Cloning and Expression of Soluble Recombinant HIV-1 CRF35 Protease-HP Thioredoxin Fusion Protein.

    Science.gov (United States)

    Azarnezhad, Asaad; Sharifi, Zohreh; Seyedabadi, Rahmatollah; Hosseini, Arshad; Johari, Behrooz; Sobhani Fard, Mahsa

    2016-01-01

    As a drug target and an antigenic agent, HIV-1 protease (HIV-1 PR) is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in E. coli and investigation of its immunoreactivity. Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning. Protease coding frame was isolated by PCR and cloned in pET102/D. TOPO expression vector and cloned protease was expressed in Escherichia coli ( E. coli) BL21 . Produced recombinant protein was purified by affinity Ni-NTA column and protein concentration was checked by BCA protein assay kit. Subsequently, immunoreactivity of recombinant protease (rPR) was assayed by Western blotting and ELISA. Cloning of the HIV protease by TOPO cloning system in pET102/D.TOPO was confirmed with PCR and sequencing. The concentration range of purified recombinant protein was 85 to 100 μg/ml . Immunogenicity of rPR was confirmed by Western blotting and ELISA. Soluble production of recombinant HIV-1 protease (HIV-1 rPR) was performed successfully. This recombinant protein disclosed 86% specificity and 90% sensitivity in immunoassay tests.

  8. Clonal diversity and estimation of relative clone age: application to agrobiodiversity of yam (Dioscorea rotundata).

    Science.gov (United States)

    Scarcelli, Nora; Couderc, Marie; Baco, Mohamed N; Egah, Janvier; Vigouroux, Yves

    2013-11-13

    Clonal propagation is a particular reproductive system found in both the plant and animal kingdoms, from human parasites to clonally propagated crops. Clonal diversity provides information about plant and animal evolutionary history, i.e. how clones spread, or the age of a particular clone. In plants, this could provide valuable information about agrobiodiversity dynamics and more broadly about the evolutionary history of a particular crop. We studied the evolutionary history of yam, Dioscorea rotundata. In Africa, Yam is cultivated by tuber clonal propagation. We used 12 microsatellite markers to identify intra-clonal diversity in yam varieties. We then used this diversity to assess the relative ages of clones. Using simulations, we assessed how Approximate Bayesian Computation could use clonal diversity to estimate the age of a clone depending on the size of the sample, the number of independent samples and the number of markers. We then applied this approach to our particular dataset and showed that the relative ages of varieties could be estimated, and that each variety could be ranked by age. We give a first estimation of clone age in an approximate Bayesian framework. However the precise estimation of clone age depends on the precision of the mutation rate. We provide useful information on agrobiodiversity dynamics and suggest recurrent creation of varietal diversity in a clonally propagated crop.

  9. Cloning of the fish cell line SSN-1 for piscine nodaviruses.

    Science.gov (United States)

    Iwamoto, T; Nakai, T; Mori, K; Arimoto, M; Furusawa, I

    2000-11-14

    Six cell clones were derived from the SSN-1 cell line, which is composed of a mixed cell population and persistently infected with a C-type retrovirus (SnRV). These clones were susceptible to 4 piscine nodavirus strains belonging to different genotypes (SJNNV, RGNNV, TPNNV and *BFNNV [striped jack, redspotted grouper, tiger puffer and barfin flounder nervous necrosis viruses]). Three clones, designated A-6, E-9, and E-11, were highly permissive to nodavirus infection and production. The virus-induced cytopathic effects appeared as cytoplasmic vacuoles and intensive disintegration at 3 to 5 d post-incubation. These observations were highly reproducible and formed the basis for a successful virus titration system. Quantitative analysis using the cloned E-11 cell line clearly revealed differences in the optimal growth temperatures among the 4 genotypic variants: 25 to 30 degrees C for strain SGWak97 (RGNNV), 20 to 25 degrees C for strain SJNag93 (SJNNV), 20 degrees C for strain TPKag93 (TPNNV), and 15 to 20 degrees C for strain JFIwa98 (BFNNV). Electron microscopy demonstrated SnRV retrovirus particles only in A-6 and E-9 cells, but PCR amplification for the pol gene and LTR region of the proviral DNA indicated the presence of the retrovirus in the other clones, including E-11. The cell clones obtained in the present study will be more useful for qualitative and quantitative analyses of piscine nodaviruses than the SSN-1 cell line.

  10. Dogs cloned from adult somatic cells.

    Science.gov (United States)

    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.

  11. Human reproductive cloning and reasons for deprivation.

    Science.gov (United States)

    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.

  12. Whole genome comparison of donor and cloned dogs.

    Science.gov (United States)

    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.

  13. Cloning arbuscule-related genes from mycorrhizas

    DEFF Research Database (Denmark)

    Burleigh, Stephen

    2000-01-01

    Until recently little was known about the identity of the genes expressed in the arbuscules of mycorrhizas, due in part to problems associated with cloning genes from the tissues of an obligate symbiont. However, the combination of advanced molecular techniques, innovative use of the materials...... available and fortuitous cloning has resulted in the recent identification of a number of arbuscule-related genes. This article provides a brief summary of the genes involved in arbuscule development, function and regulation, and the techniques used to study them. Molecular techniques include differential...

  14. Ion pair-reversed phase HPLC approach facilitates subcloning of PCR products and screening of recombinant colonies.

    Science.gov (United States)

    Shaw-Bruha, C M; Lamb, K A

    2000-04-01

    The isolation of a single DNA molecule for cloning is technically difficult, and the subsequent screening of colonies for recombinant DNA clones is time consuming. Ion pair-reversed phase HPLC (IP-RP-HPLC) analysis of nucleic acids improves the resolution and isolation of PCR products to be cloned. We demonstrate that PCR products analyzed and collected using the IP-RP-HPLC approach (WAVE DNA Fragment Analysis System) can be cloned directly into a plasmid vector. In addition, we demonstrate that when IP-RP-HPLC analysis is extended to the colony screening process, the time required for these procedures is reduced.

  15. Systemic or Intra-Amygdala Infusion of the Benzodiazepine, Midazolam, Impairs Learning, but Facilitates Re-Learning to Inhibit Fear Responses in Extinction

    Science.gov (United States)

    Hart, Genevra; Harris, Justin A.; Westbrook, R. Frederick

    2010-01-01

    A series of experiments used rats to study the effect of a systemic or intra-amygdala infusion of the benzodiazepine, midazolam, on learning and re-learning to inhibit context conditioned fear (freezing) responses. Rats were subjected to two context-conditioning episodes followed by extinction under drug or vehicle, or to two cycles of context…

  16. A web-based system to facilitate local, systematic quality improvement by multidisciplinary care teams: development and first experiences of CARDSS Online.

    Science.gov (United States)

    van Engen-Verheul, Mariëtte M; van der Veer, Sabine N; de Keizer, Nicolette F; Tjon Sjoe Sjoe, Winston; van der Zwan, Eric P A; Peek, Niels

    2013-01-01

    Continuous monitoring and systematic improvement of quality have become increasingly common in healthcare. To support multidisciplinary care teams in improving their clinical performance using feedback on quality indicators, we developed the CARDSS Online system. This system supports (i) monitoring of indicator-based performance, (ii) selecting aspects of care that need improvement, (iii) developing a quality improvement (QI) plan, and (iv) periodically adjusting the QI plan. During educational outreach visits, the system actively involves the team in the improvement effort, and guides them through the process of systematic QI without needing extensive knowledge of the underlying concepts. During the implementation of the system in the field of cardiac rehabilitation (CR) in the Netherlands, we have conducted the first outreach visits to four CR teams. During the visits, the teams formulated QI plans consisting of 4 to 7 improvement goals, each goal accompanied by 1 to 5 QI actions. Currently, we are evaluating the effect of CARDSS Online on the quality of CR in the Netherlands in a cluster randomized trial.

  17. GIS-facilitated spatial narratives

    DEFF Research Database (Denmark)

    Møller-Jensen, Lasse; Jeppesen, Henrik; Kofie, Richard Y.

    2008-01-01

    The paper examines the implementation and dissemination of spatial narratives within a geographical information system context with specific reference to portable GPS-enabled devices. The overall goal is to establish a framework for the creation of computer assisted 'guided tours' based on the th......The paper examines the implementation and dissemination of spatial narratives within a geographical information system context with specific reference to portable GPS-enabled devices. The overall goal is to establish a framework for the creation of computer assisted 'guided tours' based...... on the thematically and narrative linking of a set of locations within an area. A spatial narrative that describes the - largely unsuccessful - history of Danish plantations on the Gold Coast (1788-1850) is implemented through the Google Earth client. This client is seen both as a type of media in itself for ‘home......-based' exploration of sites related to the narrative and as a tool that facilitates the design of spatial narratives before implementation within portable GIS devices. The Google Earth-based visualization of the spatial narrative is created by a Python script that outputs a web-accessible KML format file. The KML...

  18. Some factors facilitating efficient Chinese teacher training

    OpenAIRE

    Popova, Aleksandra

    2012-01-01

    The article is devoted to the problem of Chinese teacher training major. The factors facilitating efficient Chinese teacher training as well as the most common linguistic difficulties of the Chinese language are specified in the article. The author touches upon the peculiarities of the realization of the proposed model aimed at efficient educational success within the Ukraine’s system of education.

  19. Quick and clean cloning: a ligation-independent cloning strategy for selective cloning of specific PCR products from non-specific mixes.

    Directory of Open Access Journals (Sweden)

    Frank Thieme

    Full Text Available We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplification of the insert. The other side of the linearized cloning vector has homology with a sequence present in the insert, but nested and non-overlapping with the gene-specific primer used for amplification. Since only specific products contain this sequence, but none of the non-specific products, only specific products can be cloned. Cloning is performed using a one-step reaction that only requires incubation for 10 minutes at room temperature in the presence of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. The reaction mix is then directly transformed into E. coli where the annealed vector-insert complex is repaired and ligated. We have tested this method, which we call quick and clean cloning (QC cloning, for cloning of the variable regions of immunoglobulins expressed in non-Hodgkin lymphoma tumor samples. This method can also be applied to identify the flanking sequence of DNA elements such as T-DNA or transposon insertions, or be used for cloning of any PCR product with high specificity.

  20. SysBioCube: A Data Warehouse and Integrative Data Analysis Platform Facilitating Systems Biology Studies of Disorders of Military Relevance.

    Science.gov (United States)

    Chowbina, Sudhir; Hammamieh, Rasha; Kumar, Raina; Chakraborty, Nabarun; Yang, Ruoting; Mudunuri, Uma; Jett, Marti; Palma, Joseph M; Stephens, Robert

    2013-01-01

    SysBioCube is an integrated data warehouse and analysis platform for experimental data relating to diseases of military relevance developed for the US Army Medical Research and Materiel Command Systems Biology Enterprise (SBE). It brings together, under a single database environment, pathophysio-, psychological, molecular and biochemical data from mouse models of post-traumatic stress disorder and (pre-) clinical data from human PTSD patients.. SysBioCube will organize, centralize and normalize this data and provide an access portal for subsequent analysis to the SBE. It provides new or expanded browsing, querying and visualization to provide better understanding of the systems biology of PTSD, all brought about through the integrated environment. We employ Oracle database technology to store the data using an integrated hierarchical database schema design. The web interface provides researchers with systematic information and option to interrogate the profiles of pan-omics component across different data types, experimental designs and other covariates.