Science.gov

Sample records for innovative cloning method

  1. Piglets born from handmade cloning, an innovative cloning method without micromanipulation.

    PubMed

    Du, Y; Kragh, P M; Zhang, Y; Li, J; Schmidt, M; Bøgh, I B; Zhang, X; Purup, S; Jørgensen, A L; Pedersen, A M; Villemoes, K; Yang, H; Bolund, L; Vajta, G

    2007-11-01

    Porcine handmade cloning (HMC), a simplified alternative of micromanipulation based traditional cloning (TC) has been developed in multiple phases during the past years, but the final evidence of its biological value, births of piglets was missing. Here we report the first births of healthy piglets after transfer of blastocysts produced by HMC. As a cumulative effect of technical optimization, 64.3+/-2.3 (mean+/-S.E.M.) reconstructed embryos from 151.3+/-4.8 oocytes could be obtained after 3-4h manual work, including 1h pause between fusion and activation. About half (50.1+/-2.8%, n=16) of HMC reconstructed embryos developed to blastocysts with an average cell number of 77+/-3 (n=26) after 7 days in vitro culture (IVC). According to our knowledge, this is the highest in vitro developmental rate after porcine somatic cell nuclear transfer (SCNT). A total of 416 blastocysts from HMC, mixed with 150 blastocysts from TC using a cell line from a different breed were transferred surgically to nine synchronized recipients. Out of the four pregnancies (44.4%) two were lost, while two pregnancies went to term and litters of 3 and 10 piglets were delivered by Caesarean section, with live birth/transferred embryo efficiency of 17.2% (10/58) for HMC. Although more in vivo experiments are still needed to further stabilize the system, our data proves that porcine HMC may result in birth of healthy offspring. Future comparative examinations are required to prove the value of the new technique for large-scale application.

  2. Method for cloning genes

    SciTech Connect

    Weissman, S.M.; Pereira, D.; Sood, A.

    1988-04-19

    This patent describes a recombinant cloning vehicle comprising an inserted human gene, the improvement wherein the cloning vehicle is isolated from a recombinant clone which is isolated and identified by a process comprising the steps of: (a) effecting cDNA synthesis on a mixture of mRNAs containing a target mRNA coding for a major hisitocompatibility antigen, and isolating the resultant cDNA mixture; (b) inserting the resultant cDNA into recombinant cloning vehicles, and transforming hosts with the vehicles; and (c) separating the transformants and isolating and identifying a recombinant clone containing a DNA segment which is homologous over at least a portion thereof to at least one oligonucleotide probe specific for the DNA segment.

  3. FX cloning: a simple and robust high-throughput cloning method for protein expression.

    PubMed

    Geertsma, Eric R

    2014-01-01

    The immense amount of gene sequences available nowadays allows scientist to screen broadly for extraordinary proteins. Reliable cloning tools that allow the parallel processing of many targets are vital for the success of this strategy. The FX cloning procedure detailed here is such a straightforward and efficient tool. It is dedicated to the cloning of open reading frames (ORFs) with the final aim of expressing the corresponding proteins. FX cloning combines attractive features of established high-throughput cloning methods that were thus far not unified in one single method. It facilitates the subcloning of a sequence-verified ORF to a variety of expression vectors, but is sufficiently versatile to accept PCR products as well. Moreover, the common, but undesirable feature of extending target ORFs with long cloning-related sequences is avoided. It leads to the addition of only one amino acid to each side of the protein. As a consequence, only one primer pair or PCR product suffices to generate expression vectors for both N- and C-terminal translational fusions. FX cloning is highly efficient and economical in its use. The method is suited for high-throughput cloning projects and also for everyday cloning of single targets. FX cloning is based on the use of type IIS restriction enzymes and negative selection markers. The full procedure takes place in one pot in less than 3 h and does not require intermediate purification steps nor extensive handling. The method has proven to be very robust and suitable for all common expression systems.

  4. Innovative sludge stabilization method

    SciTech Connect

    Riggenbach, J.D.

    1995-06-01

    Sludge is generated in many water and wastewater treatment processes, both biological and physical/chemical. Examples include biological sludges from sanitary and industrial wastewater treatment operations and chemical sludges such as those produced when metals are removed from metal plating wastewater. Even some potable water plants produce sludge, such as when alum is used as a flocculating agent to clarify turbid water. Because sludge is produced from such a variety of operations, different techniques have been developed to remove water from sludges and reduce the sludge volume and mass, thus making the sludge more suitable for recovery or disposal. These techniques include mechanical (e.g., filter presses), solar (sludge drying beds), and thermal. The least expensive of these methods, neglecting land costs, involves sludge drying beds and lagoons. The solar method was widely used in sewage treatment plants for many years, but has fallen in disfavor in the US; mechanical and thermal methods have been preferred. Since environmental remediation often requires managing sludges, this article presents a discussion of a variation of sludge lagoons known as evaporative sludge stabilization. Application of this process to the closure of two 2.5 acre (10117 m{sup 2}) hazardous waste surface impoundments will be discussed. 1 ref., 2 figs.

  5. Cloning cattle: the methods in the madness.

    PubMed

    Oback, Björn; Wells, David N

    2007-01-01

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

  6. [A brief introduction to the methods for novel gene cloning].

    PubMed

    Sun, C X; Yu, A C

    2000-01-01

    There are a lot of methods for novel gene cloning, but how to clone candidate gene(s) quickly and correctly? This is a brief introduction to methods of novel gene cloning, these methods includes: differential display reverse transcriptase polymerase chain reaction(DD RT-PCR), suppression subtractive hybridization(SSH), RNA arbitrarily primed PCR(RAP-PCR), representational difference analysis(RDA), yeast two-hybrid system, cDNA capturation, et al. We not only introduced these methods, but also discussed the advantages and disadvantages of them. However, no single method is omnipotent, one should pick up the method most suitable for a special purpose. PMID:12532765

  7. Innovative methods for knowledge transfer

    NASA Astrophysics Data System (ADS)

    Goanta, A. M.

    2015-11-01

    At this time, learning takes place, either with classic books on paper support or using books scanned or drawn and further converted into PDF or PPT files that are printed on type support CD / DVD. The latter modern means of learning, the study adds live on the Internet using search engines and not least e-learning method, which allows the study of bibliographic related materials in PDF or PPT, stacked and grouped on the basis of a curriculum imposed which can be accessed on a website via a user name and password. Innovative methods come to successfully use other file types than those mentioned above. The graphics in teaching technical subjects such as descriptive geometry can be achieved using animated PowerPoint files, allowing for visualization of steps to be taken, in the case of solving by drawing a descriptive geometry. Another innovative method relies on the use of HTML files, inspired by related sites help design software packages that can be used when teaching descriptive geometry that the technical design. Through this work, the author has proposed to present a new innovative method, which is inspired by the methods listed above, but involves using AVI files to teaching of computer-assisted type graphics or info graphics. In general this new author's method lends itself particularly well to the teaching of the use of software packages because the student actually see the place from where the delivered command is accessed and contextual options of right button of the mouse. These laboratory courses or mentoring can be freestanding cards support CD / DVD or can be posted on websites with restricted access based on user name and password. Practically paper presents the methodology of creating courses and tutorials in AVI format, and how to monitor the degree of accessing the website, on which there were posted mentioned teaching materials, using the tools offered by Google Analytics. The results consist of graphical work, about the degree of access to

  8. A ligation-independent cloning method using nicking DNA endonuclease.

    PubMed

    Yang, Jie; Zhang, Zhihong; Zhang, Xin A; Luo, Qingming

    2010-11-01

    Using nicking DNA endonuclease (NiDE), we developed a novel technique to clone DNA fragments into plasmids. We created a NiDE cassette consisting of two inverted NiDE substrate sites sandwiching an asymmetric four-base sequence, and NiDE cleavage resulted in 14-base single-stranded termini at both ends of the vector and insert. This method can therefore be used as a ligation-independent cloning strategy to generate recombinant constructs rapidly. In addition, we designed and constructed a simple and specific vector from an Escherichia coli plasmid back-bone to complement this cloning method. By cloning cDNAs into this modified vector, we confirmed the predicted feasibility and applicability of this cloning method. PMID:21091446

  9. Cloning

    MedlinePlus

    ... copies of whole animals Therapeutic cloning, which creates embryonic stem cells. Researchers hope to use these cells to grow healthy tissue to replace injured or diseased tissues in the human body. NIH: National Human Genome Research Institute

  10. Tissue-Culture Method of Cloning Rubber Plants

    NASA Technical Reports Server (NTRS)

    Ball, E. A.

    1983-01-01

    Guayule plant, a high-yield rubber plant cloned by tissue-culture method to produce multiple new plants that mature quickly. By adjusting culture medium, excised shoot tip produces up to 50 identical guayule plants. Varying concentration of cytokinin, single excised tip produces either 1 or several (up to 50) new plants.

  11. Outstanding Examples of Innovative Methods.

    ERIC Educational Resources Information Center

    Fink, David R., Jr.

    1983-01-01

    The author describes a conference on exploring some educational methods that have proved effective in other fields and at other levels of medical education to see if they have application to continuing medical education. (SSH)

  12. Innovative Methods for Serving Rural Handicapped Children.

    ERIC Educational Resources Information Center

    Latham, Glenn; Burnham, Julia

    1985-01-01

    Some innovative methods of serving the rural handicapped population are described: volunteers; telecommunications for home instruction; SPECIALNET electronic mail; and resources for parents. Three cases involving Utah's Cooperative Extension Project for the Handicapped summarize services for handicapped and Indian students. (GDC)

  13. Mark Making: Methodologies and methods (innovative practice).

    PubMed

    Zeilig, Hannah

    2016-09-01

    Mark Making is a recently completed AHRC-funded review exploring the role of the participative arts for people with dementia in the UK. Key concerns underlying Mark Making were both how to privilege the views and feelings of people with a dementia and also how best to understand the value of the arts for people with a dementia. These issues were tackled using a variety of qualitative methods. Methods included a rigorous literature review, the development of a unique web-based map locating many participative arts projects and above all working with people with a dementia to ascertain their views. This brief article will concentrate on some of the innovative methods that the Mark Making team used, with particular reference to comics as a mode of engagement as used in the Descartes project. The article will provide an insight into some of the methodological challenges confronted by Mark Making as well as the inspirations and successes that were enjoyed.

  14. A general one-step method for the cloning of PCR products.

    PubMed

    Bolchi, Angelo; Ottonello, Simone; Petrucco, Stefania

    2005-12-01

    A very fast, highly efficient, versatile and low-cost cloning of PCR products is described. PCR amplicons, obtained with any set of primers, is directly integrated into circular plasmid vectors by means of a one-step restriction-ligation procedure. When using proof-reading DNA polymerases, 100% cloning efficiency is easily achieved, implying that direct cloning into 'final-use' vectors (i.e. avoiding any intermediate cloning step) is a feasible task. Albeit with a lower efficiency, the same procedure is also suitable for the cloning of PCR products generated by 'non-proof-reading' DNA polymerases. Furthermore, with a simple modification of the vector polylinker site, the present method can be easily adapted to the directional cloning of open-reading-frame-encoding amplicons. This one-step procedure thus couples high efficiency with high reliability and versatility, and lends itself as the method of choice for routine cloning of PCR products.

  15. A general one-step method for the cloning of PCR products.

    PubMed

    Bolchi, Angelo; Ottonello, Simone; Petrucco, Stefania

    2005-12-01

    A very fast, highly efficient, versatile and low-cost cloning of PCR products is described. PCR amplicons, obtained with any set of primers, is directly integrated into circular plasmid vectors by means of a one-step restriction-ligation procedure. When using proof-reading DNA polymerases, 100% cloning efficiency is easily achieved, implying that direct cloning into 'final-use' vectors (i.e. avoiding any intermediate cloning step) is a feasible task. Albeit with a lower efficiency, the same procedure is also suitable for the cloning of PCR products generated by 'non-proof-reading' DNA polymerases. Furthermore, with a simple modification of the vector polylinker site, the present method can be easily adapted to the directional cloning of open-reading-frame-encoding amplicons. This one-step procedure thus couples high efficiency with high reliability and versatility, and lends itself as the method of choice for routine cloning of PCR products. PMID:15992357

  16. Generation of DNA Constructs Using the Golden GATEway Cloning Method.

    PubMed

    Kirchmaier, Stephan; Lust, Katharina; Wittbrodt, Jochen

    2017-01-01

    One of the most frequently executed tasks for molecular biologists is the design and generation of complex DNA constructs. Recently, we established the Golden GATEway cloning kit for the fast and efficient generation of transgenesis vectors. This cloning kit allows the modular assembly of DNA fragments in a defined order. The modularity reflects how complex transgenesis constructs are set up. For example, genome modification tools such as the Cre-Lox system utilize small recombination elements that are combined with larger open reading frames and noncoding regulatory DNA. Another example is that proteinogenic genes can be extended with different localisation tags or fluorescent markers. The Golden GATEway cloning kit allows focusing on the design of a transgenesis construct without having to compromise it by so far available cloning strategies. Here, we provide a step-by-step introduction on how to use the Golden GATEway cloning kit. PMID:27671939

  17. Restriction-ligation-free (RLF) cloning: a high-throughput cloning method by in vivo homologous recombination of PCR products.

    PubMed

    Wang, Y; Liu, Y; Chen, J; Tang, M J; Zhang, S L; Wei, L N; Li, C H; Wei, D B

    2015-01-01

    In this study, we optimized a restriction-ligation-free (RLF) method to save time and cost of constructing multiple plasmids with the same gene insert, and examined the efficacy of RLF on high-throughput multi-plasmid cloning. This method utilizes the precise DNA repair and recombination systems within Escherichia coli, which allows to bypass the in vitro restriction and ligation enzyme reactions commonly included in routine cloning procedures. A homologous arm is linked to the 5'-end of the forward primer used to amplify both the target gene and vector. A different homologous arm is linked to the 5'-end of the reverse primer. Therefore, genes can be cloned into the vectors by homologous recombination after co-transformation of the amplified target gene and the linearized vector, which bear the same homologous arm on either end. More than twenty-four different plasmids were generated by this method, which uses two simple polymerase chain reaction steps. This method is highly efficient in cloning any gene of interest into any vector at any site without sequence constraints, as no restriction and ligation reactions are required. PMID:26505379

  18. An accurate clone-based haplotyping method by overlapping pool sequencing

    PubMed Central

    Li, Cheng; Cao, Changchang; Tu, Jing; Sun, Xiao

    2016-01-01

    Chromosome-long haplotyping of human genomes is important to identify genetic variants with differing gene expression, in human evolution studies, clinical diagnosis, and other biological and medical fields. Although several methods have realized haplotyping based on sequencing technologies or population statistics, accuracy and cost are factors that prohibit their wide use. Borrowing ideas from group testing theories, we proposed a clone-based haplotyping method by overlapping pool sequencing. The clones from a single individual were pooled combinatorially and then sequenced. According to the distinct pooling pattern for each clone in the overlapping pool sequencing, alleles for the recovered variants could be assigned to their original clones precisely. Subsequently, the clone sequences could be reconstructed by linking these alleles accordingly and assembling them into haplotypes with high accuracy. To verify the utility of our method, we constructed 130 110 clones in silico for the individual NA12878 and simulated the pooling and sequencing process. Ultimately, 99.9% of variants on chromosome 1 that were covered by clones from both parental chromosomes were recovered correctly, and 112 haplotype contigs were assembled with an N50 length of 3.4 Mb and no switch errors. A comparison with current clone-based haplotyping methods indicated our method was more accurate. PMID:27095193

  19. Successful Innovative Methods in Introducing Astronomy Courses

    NASA Astrophysics Data System (ADS)

    Chattejee, T. K. C.

    2006-08-01

    Innovating new informative methods to induce interest in students has permitted us to introduce astronomy in several universities and institutes in Mexico. As a prelude, we gave a popular course in the history of astronomy. This was very easy as astronomy seems to be the most ancient of sciences and relating the achievements of the ancient philosophers/scientists was very enlightening. Then we put up an amateur show of the sky every week (subject to climatic conditions for observability). We showed how to take photographs and make telescopic observations. We enlightened the students of the special missions of NASA and took them to museums for space exploration. We gave a popular seminar on "Astrodynamics," highlighting its importance. We gave a series of introductory talks in radio and T.V. Finally we exposed them to electronic circulars, like "Universe Today" and "World Science." The last mentioned strategy had the most electrifying effect. We may not have been successful without it, as the students began to take the matter seriously only after reading numerous electronic circulars. In this respect, these circulars are not only informative about the latest news in astronomy, but highlight the role of astronomy in the modern world. Without it, students seem to relate astronomy to astrology; it is due to this misconception that they are not attracted to astronomy. Students were hardly convinced of the need for an astronomy course, as they did not know about the scope and development of the subject. This awakened the interests of students and they themselves proposed the initiation of an elementary course in astronomy to have a feel of the subject. Later on they proposed a course on "Rocket Dynamics." We will discuss our methods and their impact in detail.

  20. Evaluation of seamless ligation cloning extract preparation methods from an Escherichia coli laboratory strain.

    PubMed

    Okegawa, Yuki; Motohashi, Ken

    2015-10-01

    Seamless ligation cloning extract (SLiCE) is a simple and efficient method for DNA cloning without the use of restriction enzymes. Instead, SLiCE uses homologous recombination activities from Escherichia coli cell lysates. To date, SLiCE preparation has been performed using an expensive commercially available lytic reagent. To expand the utility of the SLiCE method, we evaluated different methods for SLiCE preparation that avoid using this reagent. Consequently, cell extracts prepared with buffers containing Triton X-100, which is a common and low-cost nonionic detergent, exhibited sufficient cloning activity for seamless gene incorporation into a vector.

  1. Innovating Method of Existing Mechanical Product Based on TRIZ Theory

    NASA Astrophysics Data System (ADS)

    Zhao, Cunyou; Shi, Dongyan; Wu, Han

    Main way of product development is adaptive design and variant design based on existing product. In this paper, conceptual design frame and its flow model of innovating products is put forward through combining the methods of conceptual design and TRIZ theory. Process system model of innovating design that includes requirement analysis, total function analysis and decomposing, engineering problem analysis, finding solution of engineering problem and primarily design is constructed and this establishes the base for innovating design of existing product.

  2. Cross-screening: a new method to assemble clones rapidly and unambiguously into contigs.

    PubMed

    Locke, J; Rairdan, G; McDermid, H; Nash, D; Pilgrim, D; Bell, J; Roy, K; Hodgetts, R

    1996-02-01

    We have developed a new procedure that relies on an array of cross-hybridization tests to order a set of random clones into a contig. The method, called cross-screening, uses each clone as a target and its end sequences as probes, in a matrix of reciprocal cross-hybridization tests performed on a single blot. The relationships among the clones are determined rapidly from the pairwise tests, allowing clone order to be determined directly. We have applied this technique to DNAs from a set of overlapping lambda clones from Drosophila chromosome 4. The location and orientation of each clone derived from the cross-screening data was that expected from the map assembled from overlapping restriction sites and chromosomal walking. The procedure provided additional information on a previously unknown, internally repeated DNA sequence. To demonstrate the general utility of the procedure, we have applied it to a previously described clone set within a contig in region 22q12 of human chromosome 22. The correct relative position and orientation of these clones were derived from the cross-screening data without knowledge of, or reference to, any nucleotide sequence or restriction site analysis of the DNA concerned. The cross-screening procedure is fast, economical, and robust and allows clone overlaps to be determined efficiently, with minimal interference from repeated DNA sequences. This new procedure is specifically designed for small groups of overlapping clones (tens to hundreds) and should facilitate the ordering of subclone libraries derived from small chromosomes or the large cloned inserts carried in YAC, BAC, and P1 vectors.

  3. IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation

    PubMed Central

    Wang, Jiancai; Xu, Ronghua; Liu, Aizhong

    2014-01-01

    Rapid and efficient construction of expression vectors and subsequent transformation are basic recombinant methods for the investigation of gene functionality. Although novel cloning methods have recently been developed, many laboratories worldwide continue to use traditional restriction digestion-ligation methods to construct expression vectors owing to financial constraints and the unavailability of appropriate vectors. We describe an improved restriction digestion-ligation (IRDL) cloning method that combines the advantage of directional cloning from double digestion-ligation with that of a low background observed by using a positive selection marker gene ccdB to facilitate digestion and ligation in a single tube. The IRDL cloning overcomes the time-consuming and laborious limits of traditional methods, thereby providing an easy-to-use, low-cost, and one-step strategy for directional cloning of target DNA fragments into an expression vector. As a proof-of-concept example, we developed two yeast vectors to demonstrate the feasibility and the flexibility of the IRDL cloning method. This method would provide an effective and easy-to-use system for gene cloning and functional genomics studies. PMID:25243603

  4. FIELD ANALYTICAL SCREENING PROGRAM: PCB METHOD - INNOVATIVE TECHNOLOGY REPORT

    EPA Science Inventory

    This innovative technology evaluation report (ITER) presents information on the demonstration of the U.S. Environmental Protection Agency (EPA) Region 7 Superfund Field Analytical Screening Program (FASP) method for determining polychlorinated biphenyl (PCB) contamination in soil...

  5. Cloning cattle.

    PubMed

    Oback, B; Wells, D N

    2003-01-01

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

  6. [Innovations in food processing: nonthermal methods].

    PubMed

    Herrero, A M; Romero de Avila, M D

    2006-01-01

    Consumers' desires for foods that are minimally preserved and processed are encouraging the development of new methods for the inactivation of micro-organisms in foods. Non-thermal processes (such as high hydrostatic pressure, ultrasound, irradiation, high intensity pulsed electric fields, oscillating magnetic fields and light pulses) can be used for the inactivation of food-spoilage micro-organisms without affecting the quality of the food. While the efficacy of many of these methods was demonstrated many years ago, technological advances are only now beginning to make possible their commercial exploitation.

  7. DNA methods: critical review of innovative approaches.

    PubMed

    Kok, Esther J; Aarts, Henk J M; Van Hoef, A M Angeline; Kuiper, Harry A

    2002-01-01

    The presence of ingredients derived from genetically modified organisms (GMOs) in food products in the market place is subject to a number of European regulations that stipulate which product consisting of or containing GMO-derived ingredients should be labeled as such. In order to maintain these labeling requirements, a variety of different GMO detection methods have been developed to screen for either the presence of DNA or protein derived from (approved) GM varieties. Recent incidents where unapproved GM varieties entered the European market show that more powerful GMO detection and identification methods will be needed to maintain European labeling requirements in an adequate, efficient, and cost-effective way. This report discusses the current state-of-the-art as well as future developments in GMO detection.

  8. Reality Research Methods: An Innovative Teaching Strategy for Generation Y

    ERIC Educational Resources Information Center

    van Eeden-Moorefield, Brad; Walsh, Christine

    2010-01-01

    Those who teach research methods courses may struggle to engage students who are anxious or apprehensive about taking the course. Thus, it becomes the obligation of the instructor to develop innovative approaches so that students overcome their apprehension, learn the material, and, it is hoped, develop a passion for research. The authors present…

  9. Methods to Enhance Reflective Behaviour in Innovation Processes

    ERIC Educational Resources Information Center

    Verdonschot, Suzanne G. M.

    2006-01-01

    Purpose: This study aims to trace methods that help to develop the reflective behaviour that is necessary for identifying and describing learning processes in organisations that focus on improvement and innovation. Design/methodology/approach: An extensive literature review results in the characteristics of reflection when reflection is used to…

  10. Construction of mutant alleles in Saccharomyces cerevisiae without cloning: overview and the delitto perfetto method.

    PubMed

    Moqtaderi, Zarmik; Geisberg, Joseph V

    2013-01-01

    Traditionally, methods for introducing specific new mutations at target loci in the yeast genome have involved the preparation of disruption or gene-replacement cassettes via multiple cloning steps. Sequences used for targeting these cassettes or integrating vectors are typically several hundred base pairs long. A variety of newer methods rely on the design of custom PCR oligonucleotides containing shorter sequence tails (∼50 nt) for targeting the locus of interest. These techniques obviate the need for cloning steps and allow construction of mutagenesis cassettes by PCR amplification. Such cassettes may be used for gene deletion, epitope tagging, or site-specific mutagenesis. The strategies differ in several ways, most notably with respect to whether they allow reuse of the selection marker and whether extra sequences are left behind near the target locus. This unit presents a summary of methods for targeted mutagenesis of Saccharomyces cerevisiae loci without cloning, including PCR-based allele replacement, delitto perfetto, and MIRAGE. Next, a protocol is provided for the delitto perfetto PCR- and oligonucleotide-based mutagenesis method, which offers particular advantages for generating several different mutant alleles of the same gene. PMID:24510296

  11. A fast and robust method to clone and functionally validate T-cell receptors.

    PubMed

    Birkholz, Katrin; Hofmann, Christian; Hoyer, Stefanie; Schulz, Birgit; Harrer, Thomas; Kämpgen, Eckhart; Schuler, Gerold; Dörrie, Jan; Schaft, Niels

    2009-07-31

    Sequencing, cloning and functional testing of T-cell-receptor (TCR) alpha- and beta-chains from T-cell clones is often required in immunotherapy and in immunological research. However, the determination of the TCR chains by a simple PCR is not possible, since, in contrast to the 3' constant domain and untranslated region (UTR), no conserved sequences are present in the 5' region. Furthermore, subsequent functional testing of cloned TCRs requires laborious cell culture experiments, often involving primary human material and time-consuming viral transduction strategies. Here we present a universal PCR-based protocol, adapted from the capswitch technology, that allows for amplification of the TCR alpha- and beta-chain mRNAs without knowledge of the TCR variable domain subtype by attaching a designed sequence to the mRNA's 5' end. Two different MelanA/HLA-A2-specific and one HIVgag/HLA-A2-specific TCR were cloned that way, and were functionally tested by a newly developed easy, fast, and low-cost method: we electroporated Jurkat T cells simultaneously with TCR-encoding RNA and an NFAT-reporter construct, and measured the activation status of the cells upon specific stimulation. The results of this assay correlated with the cytokine release, functional avidity, proliferative activity, and the ability to recognize MelanA/HLA-A2-presenting tumor cells of bulk T cells electroporated with RNA encoding the same TCR. Together these two protocols represent a rapid and low-cost tool for the identification and functional testing of TCRs of T-cell clones, which can then be applied in immunotherapy or immunological research. PMID:19427315

  12. Geothermal Energy Production With Innovative Methods Of Geothermal Heat Recovery

    SciTech Connect

    Swenson, Allen; Darlow, Rick; Sanchez, Angel; Pierce, Michael; Sellers, Blake

    2014-12-19

    The ThermalDrive™ Power System (“TDPS”) offers one of the most exciting technological advances in the geothermal power generation industry in the last 30 years. Using innovations in subsurface heat recovery methods, revolutionary advances in downhole pumping technology and a distributed approach to surface power production, GeoTek Energy, LLC’s TDPS offers an opportunity to change the geothermal power industry dynamics.

  13. A simple and rapid method for cloning insect vitellogenin cDNAs.

    PubMed

    Lee, J M; Hatakeyama, M; Oishi, K

    2000-03-01

    We describe a simple and rapid method for cloning insect vitellogenin (Vg) cDNAs. The method relies on the facts that insect Vg amino acid sequences can be aligned confidently along their entire lengths and that a short, highly conserved GL/ICG motif and up to nine cysteine residues that follow at conserved locations are present near the C-termini. An adaptor-ligated double-strand cDNA library is constructed from poly(A)+ RNA prepared from vitellogenic female fat body tissues using a commercial kit, and subjected to PCR with each of the degenerate nucleotide sequences for the GL/ICG motif and the adaptor sequence as primers. The PCR products (0.7-0.9 kb, representing the 3' portion) are cloned, the nucleotide sequences are determined, and the deduced amino acid sequences are aligned with the known insect Vg sequences starting from the GL/ICG motif. Gene-specific primers corresponding to the sequences near the 5'-termini of the initial clones and the adaptor sequence are employed to obtain the remaining 5' portion of the Vg cDNAs. The method was successfully applied to the bean bug Plautia stali (Heteroptera), revealing three Vg genes.

  14. Prescribing Activities that Engage Passive Residents. An Innovative Method

    PubMed Central

    Kolanowski, Ann; Buettner, Linda

    2009-01-01

    Individuals with dementia are often passive, which places them at risk for further cognitive and functional decline. Recreational activities have been used in research to reduce passive behaviors, but systematic reviews of these studies have found modest effect sizes for many activities. In this article, we describe the further theoretical development of an innovative method for prescribing activities that have a high likelihood of engaging nursing home residents who are passive and present examples for research application and clinical practice. This method may increase the effect size of activity interventions and encourage more widespread adoption of nonpharmacological interventions in practice. PMID:18274300

  15. Hot Fusion: an efficient method to clone multiple DNA fragments as well as inverted repeats without ligase.

    PubMed

    Fu, Changlin; Donovan, William P; Shikapwashya-Hasser, Olga; Ye, Xudong; Cole, Robert H

    2014-01-01

    Molecular cloning is utilized in nearly every facet of biological and medical research. We have developed a method, termed Hot Fusion, to efficiently clone one or multiple DNA fragments into plasmid vectors without the use of ligase. The method is directional, produces seamless junctions and is not dependent on the availability of restriction sites for inserts. Fragments are assembled based on shared homology regions of 17-30 bp at the junctions, which greatly simplifies the construct design. Hot Fusion is carried out in a one-step, single tube reaction at 50 °C for one hour followed by cooling to room temperature. In addition to its utility for multi-fragment assembly Hot Fusion provides a highly efficient method for cloning DNA fragments containing inverted repeats for applications such as RNAi. The overall cloning efficiency is in the order of 90-95%.

  16. Innovators.

    ERIC Educational Resources Information Center

    NEA Today, 2001

    2001-01-01

    Describes various innovations that have been developed to enhance education. These innovations include: helping educators help at-risk students succeed; promoting high school journalism; ensuring quality online learning experiences; developing a student performing group that uses theater to address social issues; and having students design their…

  17. Characterization of nutraceuticals and functional foods by innovative HPLC methods.

    PubMed

    Corradini, Claudio; Galanti, Roberta; Nicoletti, Isabella

    2002-04-01

    In recent years there is a growing interest in food and food ingredient which may provide health benefits. Food as well as food ingredients containing health-preserving components, are not considered conventional food, but can be defined as functional food. To characterise such foods, as well as nutraceuticals specific, high sensitive and reproducible analytical methodologies are needed. In light of this importance we set out to develop innovative HPLC methods employing reversed phase narrow bore column and high-performance anion-exchange chromatographic methods coupled with pulsed amperometric detection (HPAEC-PAD), which are specific for carbohydrate analysis. The developed methods were applied for the separation and quantification of citrus flavonoids and to characterize fructooligosaccharide (FOS) and fructans added to functional foods and nutraceuticals.

  18. Auto Bone Banking: Innovative Method for Bone Preservation

    PubMed Central

    M, Desai Mohan; R, Biraris Sandeep; M, Wade Roshan

    2014-01-01

    Introduction: Bone grafting is an integral part of orthopaedic surgery; the use of bone graft is increasing consistently in traumatology and also in complex revision surgeries of hip and knee arthroplasties. Considering this fact there is a need for some way to find solution for a bone graft which has more osteoinduction, osteoconduction as well as osteogenecity and also reduced rates of graft rejection and transmission of infections. All these qualities are found in autogenous bone graft. We hereby put forward a innovative method of bone preservation by using patients own femoral head and preserving it in patients own iliac pouch and making it available for future use. Case Report: From 2008 to 2012, total 17 numbers of operated sides were included in this method; patients had femoral neck fracture, osteoarthritis or avascular necrosis of femoral head and who underwent either hemi or total hip arthroplasty. Intraoperatively the resected femoral head was preserved in iliac pouch on ipsilateral side. This integrates with the native bone and additional bone graft would be made available for future use. We did not get opportunity to use the stored auograft. Conclusion: This is very innovative concept for preserving patient’s autogenous femoral head for future use. As conventional allograft relies upon screening procedure for infections, proper storage facilities and are expensive. PMID:27298993

  19. General method for cloning amplified DNA by differential screening with genomic probes.

    PubMed Central

    Brison, O; Ardeshir, F; Stark, G R

    1982-01-01

    Mutant Syrian hamster cell lines resistant to N-(phosphonacetyl)-L-aspartate, a potent and specific inhibitor of aspartate transcarbamylase, have amplified the gene coding for the multifunctional protein (CAD) that includes this activity. The average amount of DNA amplified is approximately 500 kilobases per gene copy, about 20 times the length of the CAD gene itself. A differential screening method which uses genomic DNAs as probes was developed to isolate recombinant phage containing fragments of amplified DNA. One probe was prepared by reassociating fragments of total genomic DNA from 165-28, a mutant cell line with 190 times the wild-type complement of CAD genes, until all of the sequences repeated about 200 times were annealed and then isolating the double-stranded DNA with hydroxyapatite.This DNA was highly enriched in sequences from the entire amplified region, whereas the same sequences were very rare in DNA prepared similarly from wild-type cells. After both DNAs were labeled by nick translation, highly repeated sequences were removed by hybridization to immobilized total genomic DNA from wild-type cells. A library of cloned DNA fragments from mutant 165-28 was screened with both probes, and nine independent fragments containing about 165 kilobases of amplified DNA, including the CAD gene, have been isolated so far. These cloned DNAs can be used to study the structure of the amplified region, to evaluate the nature of the amplification event, and to investigate gene expression from the amplified DNA. For example, one amplified fragment included a gene coding for a 3.8-kilobase, cytoplasmic, polyadenylated RNA which was overproduced greatly in cells resistant to N-(phosphonacetyl)-L-aspartate. The method for cloning amplified DNA is general and can be used to evaluate the possible involvement of gene amplification in phenomena such as drug resistance, transformation, or differentiation. DNA fragments corresponding to any region amplified about 10-fold or

  20. An Action Learning Method for Increased Innovation Capability in Organisations

    ERIC Educational Resources Information Center

    Olsson, Annika; Wadell, Carl; Odenrick, Per; Norell Bergendahl, Margareta

    2010-01-01

    Product innovation in highly complex and technological areas, such as medical technology, puts high requirements on the innovation capability of an organisation. Previous research and publications have highlighted organisational issues and learning matters as important and necessary for the development of innovation capability. Action learning…

  1. CLoNe is a new method to target single progenitors and study their progeny in mouse and chick.

    PubMed

    García-Moreno, Fernando; Vasistha, Navneet A; Begbie, Jo; Molnár, Zoltán

    2014-04-01

    Cell lineage analysis enables us to address pivotal questions relating to: the embryonic origin of cells and sibling cell relationships in the adult body; the contribution of progenitors activated after trauma or disease; and the comparison across species in evolutionary biology. To address such fundamental questions, several techniques for clonal labelling have been developed, each with its shortcomings. Here, we report a novel method, CLoNe that is designed to work in all vertebrate species and tissues. CLoNe uses a cocktail of labelling, targeting and transposition vectors that enables targeting of specific subpopulations of progenitor types with a combination of fluorophores resulting in multifluorescence that describes multiple clones per specimen. Furthermore, transposition into the genome ensures the longevity of cell labelling. We demonstrate the robustness of this technique in mouse and chick forebrain development, and show evidence that CLoNe will be broadly applicable to study clonal relationships in different tissues and species.

  2. CLoNe is a new method to target single progenitors and study their progeny in mouse and chick.

    PubMed

    García-Moreno, Fernando; Vasistha, Navneet A; Begbie, Jo; Molnár, Zoltán

    2014-04-01

    Cell lineage analysis enables us to address pivotal questions relating to: the embryonic origin of cells and sibling cell relationships in the adult body; the contribution of progenitors activated after trauma or disease; and the comparison across species in evolutionary biology. To address such fundamental questions, several techniques for clonal labelling have been developed, each with its shortcomings. Here, we report a novel method, CLoNe that is designed to work in all vertebrate species and tissues. CLoNe uses a cocktail of labelling, targeting and transposition vectors that enables targeting of specific subpopulations of progenitor types with a combination of fluorophores resulting in multifluorescence that describes multiple clones per specimen. Furthermore, transposition into the genome ensures the longevity of cell labelling. We demonstrate the robustness of this technique in mouse and chick forebrain development, and show evidence that CLoNe will be broadly applicable to study clonal relationships in different tissues and species. PMID:24644261

  3. CLoNe is a new method to target single progenitors and study their progeny in mouse and chick

    PubMed Central

    García-Moreno, Fernando; Vasistha, Navneet A.; Begbie, Jo; Molnár, Zoltán

    2014-01-01

    Cell lineage analysis enables us to address pivotal questions relating to: the embryonic origin of cells and sibling cell relationships in the adult body; the contribution of progenitors activated after trauma or disease; and the comparison across species in evolutionary biology. To address such fundamental questions, several techniques for clonal labelling have been developed, each with its shortcomings. Here, we report a novel method, CLoNe that is designed to work in all vertebrate species and tissues. CLoNe uses a cocktail of labelling, targeting and transposition vectors that enables targeting of specific subpopulations of progenitor types with a combination of fluorophores resulting in multifluorescence that describes multiple clones per specimen. Furthermore, transposition into the genome ensures the longevity of cell labelling. We demonstrate the robustness of this technique in mouse and chick forebrain development, and show evidence that CLoNe will be broadly applicable to study clonal relationships in different tissues and species. PMID:24644261

  4. Governing Methods: Policy Innovation Labs, Design and Data Science in the Digital Governance of Education

    ERIC Educational Resources Information Center

    Williamson, Ben

    2015-01-01

    Policy innovation labs are emerging knowledge actors and technical experts in the governing of education. The article offers a historical and conceptual account of the organisational form of the policy innovation lab. Policy innovation labs are characterised by specific methods and techniques of design, data science, and digitisation in public…

  5. MEGAWHOP cloning: a method of creating random mutagenesis libraries via megaprimer PCR of whole plasmids.

    PubMed

    Miyazaki, Kentaro

    2011-01-01

    MEGAWHOP allows for the cloning of DNA fragments into a vector and is used for conventional restriction digestion/ligation-based procedures. In MEGAWHOP, the DNA fragment to be cloned is used as a set of complementary primers that replace a homologous region in a template vector through whole-plasmid PCR. After synthesis of a nicked circular plasmid, the mixture is treated with DpnI, a dam-methylated DNA-specific restriction enzyme, to digest the template plasmid. The DpnI-treated mixture is then introduced into competent Escherichia coli cells to yield plasmids carrying replaced insert fragments. Plasmids produced by the MEGAWHOP method are virtually free of contamination by species without any inserts or with multiple inserts, and also the parent. Because the fragment is usually long enough to not interfere with hybridization to the template, various types of fragments can be used with mutations at any site (either known or unknown, random, or specific). By using fragments having homologous sequences at the ends (e.g., adaptor sequence), MEGAWHOP can also be used to recombine nonhomologous sequences mediated by the adaptors, allowing rapid creation of novel constructs and chimeric genes. PMID:21601687

  6. Reassembling the Information Technology Innovation Process: An Actor Network Theory Method for Managing the Initiation, Production, and Diffusion of Innovations

    NASA Astrophysics Data System (ADS)

    Zendejas, Gerardo; Chiasson, Mike

    This paper will propose and explore a method to enhance focal actors' abilities to enroll and control the many social and technical components interacting during the initiation, production, and diffusion of innovations. The reassembling and stabilizing of such components is the challenging goal of the focal actors involved in these processes. To address this possibility, a healthcare project involving the initiation, production, and diffusion of an IT-based innovation will be influenced by the researcher, using concepts from actor network theory (ANT), within an action research methodology (ARM). The experiences using this method, and the nature of enrolment and translation during its use, will highlight if and how ANT can provide a problem-solving method to help assemble the social and technical actants involved in the diffusion of an innovation. Finally, the paper will discuss the challenges and benefits of implementing such methods to attain widespread diffusion.

  7. Function combined method for design innovation of children's bike

    NASA Astrophysics Data System (ADS)

    Wu, Xiaoli; Qiu, Tingting; Chen, Huijuan

    2013-03-01

    As children mature, bike products for children in the market develop at the same time, and the conditions are frequently updated. Certain problems occur when using a bike, such as cycle overlapping, repeating function, and short life cycle, which go against the principles of energy conservation and the environmental protection intensive design concept. In this paper, a rational multi-function method of design through functional superposition, transformation, and technical implementation is proposed. An organic combination of frog-style scooter and children's tricycle is developed using the multi-function method. From the ergonomic perspective, the paper elaborates on the body size of children aged 5 to 12 and effectively extracts data for a multi-function children's bike, which can be used for gliding and riding. By inverting the body, parts can be interchanged between the handles and the pedals of the bike. Finally, the paper provides a detailed analysis of the components and structural design, body material, and processing technology of the bike. The study of Industrial Product Innovation Design provides an effective design method to solve the bicycle problems, extends the function problems, improves the product market situation, and enhances the energy saving feature while implementing intensive product development effectively at the same time.

  8. Simplified methods for the construction of RNA and DNA virus infectious clones.

    PubMed

    Nagata, Tatsuya; Inoue-Nagata, Alice Kazuko

    2015-01-01

    Infectious virus clones are one of the most powerful tools in plant pathology, molecular biology, and biotechnology. The construction of infectious clones of RNA and DNA viruses, however, usually requires laborious cloning and subcloning steps. In addition, instability of the RNA virus genome is frequently reported after its introduction into the vector and transference to Escherichia coli. These difficulties hamper the cloning procedures, making it tedious and cumbersome. This chapter describes two protocols for a simple construction of infectious viruses, an RNA virus, the tobamovirus Pepper mild mottle virus, and a DNA virus, a bipartite begomovirus. For this purpose, the strategy of overlap-extension PCR was used for the construction of infectious tobamovirus clone and of rolling circle amplification (RCA) for the construction of a dimeric form of the begomovirus clone. PMID:25287508

  9. Simple and versatile molecular method of copy-number measurement using cloned competitors.

    PubMed

    Kim, Hyun-Kyoung; Hwang, Hai-Li; Park, Seong-Yeol; Lee, Kwang Man; Park, Won Cheol; Kim, Han-Seong; Um, Tae-Hyun; Hong, Young Jun; Lee, Jin Kyung; Joo, Sun-Young; Seoh, Ju-Young; Song, Yeong-Wook; Kim, Soo-Youl; Kim, Yong-Nyun; Hong, Kyeong-Man

    2013-01-01

    Variations and alterations of copy numbers (CNVs and CNAs) carry disease susceptibility and drug responsiveness implications. Although there are many molecular methods to measure copy numbers, sensitivity, reproducibility, cost, and time issues remain. In the present study, we were able to solve those problems utilizing our modified real competitive PCR method with cloned competitors (mrcPCR). First, the mrcPCR for ERBB2 copy number was established, and the results were comparable to current standard methods but with a shorter assay time and a lower cost. Second, the mrcPCR assays for 24 drug-target genes were established, and the results in a panel of NCI-60 cells were comparable to those from real-time PCR and microarray. Third, the mrcPCR results for FCGR3A and the FCGR3B CNVs were comparable to those by the paralog ratio test (PRT), but without PRT's limitations. These results suggest that mrcPCR is comparable to the currently available standard or the most sensitive methods. In addition, mrcPCR would be invaluable for measurement of CNVs in genes with variants of similar structures, because combination of the other methods is not necessary, along with its other advantages such as short assay time, small sample amount requirement, and applicability to all sequences and genes.

  10. Simple and Versatile Molecular Method of Copy-Number Measurement Using Cloned Competitors

    PubMed Central

    Kim, Hyun-Kyoung; Hwang, Hai-Li; Park, Seong-Yeol; Lee, Kwang Man; Park, Won Cheol; Kim, Han-Seong; Um, Tae-Hyun; Hong, Young Jun; Lee, Jin Kyung; Joo, Sun-Young; Seoh, Ju-Young; Song, Yeong-Wook; Kim, Soo-Youl; Kim, Yong-Nyun; Hong, Kyeong-Man

    2013-01-01

    Variations and alterations of copy numbers (CNVs and CNAs) carry disease susceptibility and drug responsiveness implications. Although there are many molecular methods to measure copy numbers, sensitivity, reproducibility, cost, and time issues remain. In the present study, we were able to solve those problems utilizing our modified real competitive PCR method with cloned competitors (mrcPCR). First, the mrcPCR for ERBB2 copy number was established, and the results were comparable to current standard methods but with a shorter assay time and a lower cost. Second, the mrcPCR assays for 24 drug-target genes were established, and the results in a panel of NCI-60 cells were comparable to those from real-time PCR and microarray. Third, the mrcPCR results for FCGR3A and the FCGR3B CNVs were comparable to those by the paralog ratio test (PRT), but without PRT's limitations. These results suggest that mrcPCR is comparable to the currently available standard or the most sensitive methods. In addition, mrcPCR would be invaluable for measurement of CNVs in genes with variants of similar structures, because combination of the other methods is not necessary, along with its other advantages such as short assay time, small sample amount requirement, and applicability to all sequences and genes. PMID:23936009

  11. Effect of preparation method on the glycaemic index of novel potato clones.

    PubMed

    Kinnear, Tara; Wolever, Thomas M S; Murphy, Agnes M; Sullivan, J Alan; Liu, Qiang; Bizimungu, Benoit

    2011-08-01

    The purpose of this study was to investigate whether the effects of cooling and reheating on the glycaemic index (GI) of novel potato clones (selections) differed depending on selection and whether cooling altered starch absorption in vivo. We conducted 3 experiments using 4 novel potato clones in healthy subjects. Experiment 1: the GI of 4 selections each prepared in 3 ways (freshly boiled, cooled, or cooled and reheated) was measured in 2 groups of 10 subjects (each group tested 2 selections). Experiment 2 (n=10): two selections from Experiment 1 were re-tested one year later, by a different subject group. Experiment 3 (n=10): two selections from Experiment 1 were tested by subjects from Experiment 2 to assess the rate and extent of starch absorption using the second-meal effect and the breath hydrogen method, respectively. Experiment 1 demonstrated a selection×treatment interaction for GI (p=0.024); cooling reduced the GI of two selections by 40-50% (p<0.05) but reduced GI of the other 2 by only 8-10% (ns). Experiment 2 confirmed the selection×treatment interaction (p=0.018) seen in Experiment 1. Experiment 3: cooling reduced the GI by an average of 37% (p<0.05) but only increased starch malabsorption in vivo from 3% to 5% (p=0.021); there was no significant second-meal effect. It is concluded that the effect of cooling on the GI of potatoes may vary from 0-50% depending on selection. However, the mechanism for the effect is not clear: the 2% increase in starch malabsorption seen upon cooling potatoes was not nearly enough to account for the 37% reduction in GI.

  12. Methods to Implement Innovation and Entrepreneurship in Physics

    NASA Astrophysics Data System (ADS)

    Arion, Douglas

    2015-03-01

    The physics community is beginning to become aware of the benefits of entrepreneurship and innovation education: greater enrollments, improved students satisfaction, a wider range of interesting research problems, and the potential for greater return from more successful alumni. This talk will suggest a variety of mechanisms by which physics departments can include entrepreneurship and innovation content within their programs - without necessarily requiring earth-shattering changes to the curriculum. These approaches will thus make it possible for departments to get involved with entrepreneurship and innovation, and grow those components into vibrant activities for students and faculty.

  13. INNOVATIVE METHODS FOR EMISSION INVENTORY DEVELOPMENT AND EVALUATION: WORKSHOP SYNTHESIS

    EPA Science Inventory

    Emission inventories are key databases for evaluating, managing, and regulating air pollutants. Refinements and innovations in instruments that measure air pollutants, models that calculate emissions, and techniques for data management and uncertainty assessment are critical to ...

  14. Influence of embryo handling and transfer method on pig cloning efficiency.

    PubMed

    Shi, Junsong; Zhou, Rong; Luo, Lvhua; Mai, Ranbiao; Zeng, Haiyu; He, Xiaoyan; Liu, Dewu; Zeng, Fang; Cai, Gengyuan; Ji, Hongmei; Tang, Fei; Wang, Qinglai; Wu, Zhenfang; Li, Zicong

    2015-03-01

    The somatic cell nuclear transfer (SCNT) technique could be used to produce genetically superior or genetically engineered cloned pigs that have wide application in agriculture and bioscience research. However, the efficiency of porcine SCNT currently is very low. Embryo transfer (ET) is a key step for the success of SCNT. In this study, the effects of several ET-related factors, including cloned embryo culture time, recipient's ovulation status, co-transferred helper embryos and ET position, on the success rate of pig cloning were investigated. The results indicated that transfer of cloned embryos cultured for a longer time (22-24h vs. 4-6h) into pre-ovulatory sows decreased recipient's pregnancy rate and farrowing rate, and use of pre-ovulatory and post-ovulatory sows as recipients for SCNT embryos cultured for 22-24h resulted in a similar porcine SCNT efficiency. Use of insemination-produced in vivo fertilized, parthenogenetically activated and in vitro fertilized embryos as helper embryos to establish and/or maintain pregnancy of SCNT embryos recipients could not improve the success rate of porcine SCNT. Transfer of cloned embryos into double oviducts of surrogates significantly increased pregnancy rate as well as farrowing rate of recipients, and the developmental rate of transferred cloned embryos, as compared to unilateral oviduct transfer. This study provided useful information for optimization of the embryo handling and transfer protocol, which will help to improve the ability to generate cloned pigs.

  15. An automated method for DNA preparation from thousands of YAC clones.

    PubMed Central

    MacMurray, A J; Weaver, A; Shin, H S; Lander, E S

    1991-01-01

    We describe an automated method for the preparation of yeast genomic DNA capable of preparing thousands of DNAs in parallel from a YAC library. Briefly, the protocol involves four steps: (1) Yeast clones are grown in the wells of 96-well microtiter plates with filter (rather than plastic) well-bottoms, which are embedded in solid growth media; (2) These yeast cultures are resuspended and their concentrations determined by optical density measurement; (3) Equal numbers of cells from each well are embedded in low-melting temperature agarose blocks in fresh 96-well plates, again with filter bottoms; and (4) DNA is prepared in the agarose blocks by a protocol similar to that used for preparing DNA for pulsed-field gels, with the reagents being dialyzed through the (filter) bottoms of the microtiter plate. The DNA produced by this method is suitable for pulsed-field gel electrophoresis, for restriction enzyme digestion, and for the polymerase chain reaction (PCR). Using this protocol, we produced 3000 YAC strain DNAs in three weeks. This automated procedure should be extremely useful in many genomic mapping projects. Images PMID:2014175

  16. Comparison of Molecular Typing Methods for Characterization of Staphylococcus epidermidis: Proposal for Clone Definition▿ †

    PubMed Central

    Miragaia, M.; Carriço, J. A.; Thomas, J. C.; Couto, I.; Enright, M. C.; de Lencastre, H.

    2008-01-01

    In the present study we give some direction on the selection of the most appropriate typing method(s) to be used for the characterization of Staphylococcus epidermidis, in view of the most recent findings on the evolution, population structure, and epidemiology of this species. In order to achieve this aim, quantitative assessment of the correlation of the results of three typing methods—pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and staphylococcal chromosomal cassette mec (SCCmec) typing, which target different regions of the chromosome that evolve at different rates—was performed. In order to evaluate the discriminatory ability and the strength and direction of the correlation of the different typing methods, Simpson's index of diversity (SID), the adjusted Rand coefficient (AR), and the Wallace coefficient (W) were calculated. PFGE was the most discriminatory method (SID = 99%), followed by MLST (SID = 90%) and SCCmec typing (SID = 75%). The values of AR and W (0.10 < AR < 0.30; 0.50 < W < 0.75) indicated that the partition of the same isolate collection by PFGE, MLST, and SCCmec typing provided results that had only a poor correlation with each other. However, the information provided by the combination of PFGE and SCCmec enabled the prediction of the results obtained by MLST at the level of the clonal complex with a high degree of precision (W > 0.90). We propose that clones of S. epidermidis be defined by the combination of the PFGE type followed by the SCCmec type, which provides reliable information on the short-term epidemiology and the ability to predict with consistency long-term clonal evolution. PMID:17989196

  17. Highly efficient and reliable chemically assisted enucleation method for handmade cloning in cattle.

    PubMed

    Vajta, Gábor; Maddox-Hyttel, Poul; Skou, Christina T; Tecirlioglu, R Tayfur; Peura, Teija T; Lai, Liangxue; Murphy, Clifton N; Prather, Randall S; Kragh, Peter M; Callesen, Henrik

    2005-01-01

    The purpose of the present study was to find an efficient and reliable chemically assisted procedure for enucleation related to the handmade cloning (HMC) technique. After in vitro maturation oocytes were incubated in 0.5 microg mL(-1) demecolcine for 2 h. Subsequently, zonae pellucidae were digested with pronase, and one-third of the cytoplasm connected to an extrusion cone was removed by hand using a microblade. The remaining two-thirds were used as recipients for HMC, and reconstructed and activated embryos were cultured for 7 days. The time-dependent manner of the development of extrusion cones, the efficiency (oriented bisection per oocyte; 94%), reliability (success per attempted enucleation; 98%), and the blastocyst per reconstructed embryo rates (48%) were measured. Ultrastructural analyses demonstrated that demecolcine treatment resulted in disoriented and haphazardly orientated microtubules. The general ultrastructure of the oocyte organelles, however, appeared to be unaltered by the treatments. Considering that no oocyte selection based on polar body presence was performed, this system seems to be more efficient and reliable than any other enucleation method. Moreover, expensive equipment (inverted fluorescence microscope) and a potentially harmful step (staining and ultraviolet illumination) can be eliminated from the HMC procedure without compromising the high in vitro efficiency.

  18. Changing Methods and Mindsets: Lessons from Innovate NYC

    ERIC Educational Resources Information Center

    Hodas, Steven

    2016-01-01

    As schools and classrooms explore technology-based, student-centered, personalized approaches to teaching and learning, their efforts to innovate can be hamstrung by archaic district operating systems that do not allow them to take advantage of new technologies, to work with smaller startup companies, or to quickly make and implement decisions.…

  19. Innovative Methods in Technological Education. Studies in Engineering Education 13.

    ERIC Educational Resources Information Center

    Fishwick, W., Ed.

    This book presents 16 papers on problems and innovations in postsecondary engineering education. An introduction discusses the major issues currently facing engineering education. Chapter 2 briefly reviews the papers which are grouped under topics. Three of thee papers comment on teaching non-technical matters and attitudes. Two papers point out…

  20. Analysis of flood modeling through innovative geomatic methods

    NASA Astrophysics Data System (ADS)

    Zazo, Santiago; Molina, José-Luis; Rodríguez-Gonzálvez, Pablo

    2015-05-01

    A suitable assessment and management of the exposure level to natural flood risks necessarily requires an exhaustive knowledge of the terrain. This study, primarily aimed to evaluate flood risk, firstly assesses the suitability of an innovative technique, called Reduced Cost Aerial Precision Photogrammetry (RC-APP), based on a motorized technology ultra-light aircraft ULM (Ultra-Light Motor), together with the hybridization of reduced costs sensors, for the acquisition of geospatial information. Consequently, this research generates the RC-APP technique which is found to be a more accurate-precise, economical and less time consuming geomatic product. This technique is applied in river engineering for the geometric modeling and risk assessment to floods. Through the application of RC-APP, a high spatial resolution image (orthophoto of 2.5 cm), and a Digital Elevation Model (DEM) of 0.10 m mesh size and high density points (about 100 points/m2), with altimetric accuracy of -0.02 ± 0.03 m have been obtained. These products have provided a detailed knowledge of the terrain, afterward used for the hydraulic simulation which has allowed a better definition of the inundated area, with important implications for flood risk assessment and management. In this sense, it should be noted that the achieved spatial resolution of DEM is 0.10 m which is especially interesting and useful in hydraulic simulations through 2D software. According to the results, the developed methodology and technology allows for a more accurate riverbed representation, compared with other traditional techniques such as Light Detection and Ranging (LiDAR), with a Root-Mean-Square Error (RMSE ± 0.50 m). This comparison has revealed that RC-APP has one lower magnitude order of error than the LiDAR method. Consequently, this technique arises as an efficient and appropriate tool, especially in areas with high exposure to risk of flooding. In hydraulic terms, the degree of detail achieved in the 3D model

  1. An Alternative Method to Facilitate cDNA Cloning for Expression Studies in Mammalian Cells by Introducing Positive Blue White Selection in Vaccinia Topoisomerase I-Mediated Recombination

    PubMed Central

    Udo, Hiroshi

    2015-01-01

    One of the most basic techniques in biomedical research is cDNA cloning for expression studies in mammalian cells. Vaccinia topoisomerase I-mediated cloning (TOPO cloning by Invitrogen) allows fast and efficient recombination of PCR-amplified DNAs. Among TOPO vectors, a pcDNA3.1 directional cloning vector is particularly convenient, since it can be used for expression analysis immediately after cloning. However, I found that the cloning efficiency was reduced when RT-PCR products were used as inserts (about one-quarter). Since TOPO vectors accept any PCR products, contaminating fragments in the insert DNA create negative clones. Therefore, I designed a new mammalian expression vector enabling positive blue white selection in Vaccinia topoisomerase I–mediated cloning. The method utilized a short nontoxic LacZα peptide as a linker for GFP fusion. When cDNAs were properly inserted into the vector, minimal expression of the fusion proteins in E. coli (harboring lacZΔM15) resulted in formation of blue colonies on X-gal plates. This method improved both cloning efficiency (75%) and directional cloning (99%) by distinguishing some of the negative clones having non-cording sequences, since these inserts often disturbed translation of lacZα. Recombinant plasmids were directly applied to expression studies using GFP as a reporter. Utilization of the P2A peptide allowed for separate expression of GFP. In addition, the preparation of Vaccinia topoisomerase I-linked vectors was streamlined, which consisted of successive enzymatic reactions with a single precipitation step, completing in 3 hr. The arrangement of unique restriction sites enabled further modification of vector components for specific applications. This system provides an alternative method for cDNA cloning and expression in mammalian cells. PMID:26422141

  2. An Alternative Method to Facilitate cDNA Cloning for Expression Studies in Mammalian Cells by Introducing Positive Blue White Selection in Vaccinia Topoisomerase I-Mediated Recombination.

    PubMed

    Udo, Hiroshi

    2015-01-01

    One of the most basic techniques in biomedical research is cDNA cloning for expression studies in mammalian cells. Vaccinia topoisomerase I-mediated cloning (TOPO cloning by Invitrogen) allows fast and efficient recombination of PCR-amplified DNAs. Among TOPO vectors, a pcDNA3.1 directional cloning vector is particularly convenient, since it can be used for expression analysis immediately after cloning. However, I found that the cloning efficiency was reduced when RT-PCR products were used as inserts (about one-quarter). Since TOPO vectors accept any PCR products, contaminating fragments in the insert DNA create negative clones. Therefore, I designed a new mammalian expression vector enabling positive blue white selection in Vaccinia topoisomerase I-mediated cloning. The method utilized a short nontoxic LacZα peptide as a linker for GFP fusion. When cDNAs were properly inserted into the vector, minimal expression of the fusion proteins in E. coli (harboring lacZΔM15) resulted in formation of blue colonies on X-gal plates. This method improved both cloning efficiency (75%) and directional cloning (99%) by distinguishing some of the negative clones having non-cording sequences, since these inserts often disturbed translation of lacZα. Recombinant plasmids were directly applied to expression studies using GFP as a reporter. Utilization of the P2A peptide allowed for separate expression of GFP. In addition, the preparation of Vaccinia topoisomerase I-linked vectors was streamlined, which consisted of successive enzymatic reactions with a single precipitation step, completing in 3 hr. The arrangement of unique restriction sites enabled further modification of vector components for specific applications. This system provides an alternative method for cDNA cloning and expression in mammalian cells. PMID:26422141

  3. General method for cloning Neurospora crassa nuclear genes by complementation of mutants.

    PubMed Central

    Akins, R A; Lambowitz, A M

    1985-01-01

    We have developed a sib selection procedure for cloning Neurospora crassa nuclear genes by complementation of mutants. This procedure takes advantage of a modified N. crassa transformation procedure that gives as many as 10,000 to 50,000 stable transformants per microgram of DNA with recombinant plasmids containing the N. crassa qa-2+ gene. Here, we describe the use of the sib selection procedure to clone genes corresponding to auxotrophic mutants, nic-1 and inl. The identities of the putative clones were confirmed by mapping their chromosomal locations in standard genetic crosses and using restriction site polymorphisms as genetic markers. Because we can obtain very high N. crassa transformation frequencies, cloning can be accomplished with as few as five subdivisions of an N. crassa genomic library. The sib selection procedure should, for the first time, permit the cloning of any gene corresponding to an N. crassa mutant for which an appropriate selection can be devised. Analogous procedures may be applicable to other filamentous fungi before the development of operational shuttle vectors. Images PMID:2942762

  4. An Innovative Cloning Platform Enables Large-Scale Production and Maturation of an Oxygen-Tolerant [NiFe]-Hydrogenase from Cupriavidus necator in Escherichia coli

    PubMed Central

    Schiffels, Johannes; Pinkenburg, Olaf; Schelden, Maximilian; Aboulnaga, El-Hussiny A. A.; Baumann, Marcus E. M.; Selmer, Thorsten

    2013-01-01

    Expression of multiple heterologous genes in a dedicated host is a prerequisite for approaches in synthetic biology, spanning from the production of recombinant multiprotein complexes to the transfer of tailor-made metabolic pathways. Such attempts are often exacerbated, due in most cases to a lack of proper directional, robust and readily accessible genetic tools. Here, we introduce an innovative system for cloning and expression of multiple genes in Escherichia coli BL21 (DE3). Using the novel methodology, genes are equipped with individual promoters and terminators and subsequently assembled. The resulting multiple gene cassettes may either be placed in one vector or alternatively distributed among a set of compatible plasmids. We demonstrate the effectiveness of the developed tool by production and maturation of the NAD+reducing soluble [NiFe]-hydrogenase (SH) from Cupriavidus necator H16 (formerly Ralstonia eutropha H16) in E. coli BL21Star™ (DE3). The SH (encoded in hoxFUYHI) was successfully matured by co-expression of a dedicated set of auxiliary genes, comprising seven hyp genes (hypC1D1E1A2B2F2X) along with hoxW, which encodes a specific endopeptidase. Deletion of genes involved in SH maturation reduced maturation efficiency substantially. Further addition of hoxN1, encoding a high-affinity nickel permease from C. necator, considerably increased maturation efficiency in E. coli. Carefully balanced growth conditions enabled hydrogenase production at high cell-densities, scoring mg·(Liter culture)−1 yields of purified functional SH. Specific activities of up to 7.2±1.15 U·mg−1 were obtained in cell-free extracts, which is in the range of the highest activities ever determined in C. necator extracts. The recombinant enzyme was isolated in equal purity and stability as previously achieved with the native form, yielding ultrapure preparations with anaerobic specific activities of up to 230 U·mg−1. Owing to the combinatorial power exhibited by the

  5. Study of the efficiency of chemically assisted enucleation method for handmade cloning in goat (Capra hircus).

    PubMed

    Akshey, Y S; Malakar, D; De, A K; Jena, M K; Sahu, S; Dutta, R

    2011-08-01

    The present investigation was carried out to find an efficient chemically assisted procedure for enucleation of goat oocytes related to handmade cloning (HMC) technique. After 22-h in vitro maturation, oocytes were incubated with 0.5 μg/ml demecolcine for 2 h. Cumulus cells were removed by pipetting and vortexing in 0.5 mg/ml hyaluronidase, and zona pellucida were digested with pronase. Oocytes with extrusion cones were subjected to oriented bisection. One-third of the cytoplasm with the extrusion cone was removed with a micro blade. The remaining cytoplasts were used as recipients in HMC. Goat foetal fibroblasts were used as nuclear donors. The overall efficiency measured as the number of cytoplasts obtained per total number of oocytes used was significantly (p < 0.05) higher in chemically assisted handmade enucleation (CAHE) than oriented handmade enucleation without demecolcine (OHE) (80.02 ± 1.292% vs. 72.9 ± 1.00%, respectively, mean ± SEM). The reconstructed and activated embryos were cultured in embryo development medium (EDM) for 7 days. Fusion, cleavage and blastocyst development rate were 71.63 ± 1.95%, 92.94 ± 0.91% and 23.78 ± 3.33% (mean ± SEM), respectively which did not differ significantly from those achieved with random handmade enucleation and OHE. In conclusion, chemically assisted enucleation is a highly efficient and reliable enucleation method for goat HMC which eliminates the need of expensive equipment (inverted fluorescence microscope) and potentially harmful chromatin staining and ultraviolet (UV) irradiation for cytoplast selection.

  6. A Rapid and Efficient Method for Construction of an Infectious Clone of Tomato yellow leaf curl virus.

    PubMed

    Bang, Bongjun; Lee, Jongyun; Kim, Sunyoung; Park, Jungwook; Nguyen, Thao Thi; Seo, Young-Su

    2014-09-01

    Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is responsible for one of the most devastating viral diseases in tomato-growing countries and is becoming a serious problem in many subtropical and tropical countries. The climate in Korea is getting warmer and developing subtropical features in response to global warming. These changes are being accompanied by TYLCV, which is now becoming a large problem in the Korean tomato industry. The most effective way to reduce damage caused by TYLCV is to breed resistant varieties of tomatoes. To accomplish this, it is necessary to establish a simple inoculation technique for the efficient evaluation of resistance to TYLCV. Here, we present the rolling circle amplification (RCA) method, which employs a bacteriophage using phi-29 DNA polymerase for construction of infectious TYLCV clones. The RCA method is simple, does not require sequence information for cloning, and is less expensive and time consuming than conventional PCR based-methods. Furthermore, RCA-based construction of an infectious clone can be very useful to other emerging and unknown geminiviruses in Korea. PMID:25289018

  7. A Rapid and Efficient Method for Construction of an Infectious Clone of Tomato yellow leaf curl virus

    PubMed Central

    Bang, Bongjun; Lee, Jongyun; Kim, Sunyoung; Park, Jungwook; Nguyen, Thao Thi; Seo, Young-Su

    2014-01-01

    Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is responsible for one of the most devastating viral diseases in tomato-growing countries and is becoming a serious problem in many subtropical and tropical countries. The climate in Korea is getting warmer and developing subtropical features in response to global warming. These changes are being accompanied by TYLCV, which is now becoming a large problem in the Korean tomato industry. The most effective way to reduce damage caused by TYLCV is to breed resistant varieties of tomatoes. To accomplish this, it is necessary to establish a simple inoculation technique for the efficient evaluation of resistance to TYLCV. Here, we present the rolling circle amplification (RCA) method, which employs a bacteriophage using phi-29 DNA polymerase for construction of infectious TYLCV clones. The RCA method is simple, does not require sequence information for cloning, and is less expensive and time consuming than conventional PCR based-methods. Furthermore, RCA-based construction of an infectious clone can be very useful to other emerging and unknown geminiviruses in Korea. PMID:25289018

  8. Generation of cloned and chimeric embryos/offspring using the new methods of animal biotechnology.

    PubMed

    Skrzyszowska, Maria; Karasiewicz, Jolanta; Bednarczyk, Marek; Samiec, Marcin; Smorag, Zdzisław; Waś, Bogusław; Guszkiewicz, Andrzej; Korwin-Kossakowski, Maciej; Górniewska, Maria; Szablisty, Ewa; Modliński, Jacek A; Łakota, Paweł; Wawrzyńska, Magdalena; Sechman, Andrzej; Wojtysiak, Dorota; Hrabia, Anna; Mika, Maria; Lisowski, Mirosław; Czekalski, Przemysław; Rzasa, Janusz; Kapkowska, Ewa

    2006-01-01

    The article summarizes results of studies concerning: 1/ qualitative evaluation of pig nuclear donor cells to somatic cell cloning, 2/ developmental potency of sheep somatic cells to create chimera, 3/ efficient production of chicken chimera. The quality of nuclear donor cells is one of the most important factors to determine the efficiency of somatic cell cloning. Morphological criteria commonly used for qualitative evaluation of somatic cells may be insufficient for practical application in the cloning. Therefore, different types of somatic cells being the source of genomic DNA in the cloning procedure were analyzed on apoptosis with the use of live-DNA or plasma membrane fluorescent markers. It has been found that morphological criteria are a sufficient selection factor for qualitative evaluation of nuclear donor cells to somatic cell cloning. Developmental potencies of sheep somatic cells in embryos and chimeric animals were studied using blastocyst complementation test. Fetal fibroblasts stained with vital fluorescent dye and microsurgically placed in morulae or blastocysts were later identified in embryos cultured in vitro. Transfer of Polish merino blastocysts harbouring Heatherhead fibroblasts to recipient ewes brought about normal births at term. Newly-born animals were of merino appearance with dark patches on their noses, near the mouth and on their clovens. This overt chimerism shows that fetal fibroblasts introduced to sheep morulae/blastocysts revealed full developmental plasticity. To achieve the efficient production of chicken chimeras, the blastodermal cells from embryos of the donor breeds, (Green-legged Partridgelike breed or GPxAraucana) were transferred into the embryos of the recipient breed (White Leghorn), and the effect of chimerism on the selected reproductive and physiological traits of recipients was examined. Using the model which allowed identification of the chimerism at many loci, it has been found that 93.9% of the examined birds

  9. Bovine oocytes vitrified by the open pulled straw method and used for somatic cell cloning supported development to term.

    PubMed

    Hou, Yun-Peng; Dai, Yun-Ping; Zhu, Shi-En; Zhu, Hua-Bin; Wu, Tong-Yi; Gong, Guo-Chun; Wang, Hai-Ping; Wang, Li-Li; Liu, Ying; Li, Rong; Wan, Rong; Li, Ning

    2005-10-01

    The objective of the present study was to determine if oocytes vitrified by the open pulled straw (OPS) method could subsequently be used to produce somatic cell cloned cattle. Post-thaw survival rates were 77.0, 79.1, 97.2 and 97.5% for oocytes vitrified with EDFS30 (15% ethylene glycol, 15% dimethyl sulfoxide, ficoll and sucrose), EDFS40 (20% ethylene glycol, 20% dimethyl sulfoxide, ficoll and sucrose), EDFSF30 (15% ethylene glycol, 15% dimethyl sulfoxide, ficoll, sucrose and FBS) and EDFSF40 (20% ethylene glycol, 20% dimethyl sulfoxide, ficoll, sucrose and FBS), respectively. The parthenogenetic blastocyst rates of the vitrified-thawed oocytes activated with 5 microM of the calcium ionophore A23187 for 5 min and 2 microM of 6-dimethylaminopurin (6-DMAP) for 4h ranged from 10.3 to 23.0%, with the highest group not significantly differing from that of the controls (33.2%). In total, 722 vitrified-thawed oocytes were used as recipients for nuclear transfer, of which 343 fused (47.6%). Fifty-six (16.3%) of the reconstructed embryos reached the blastocyst stage after 7d of in vitro culture. Twenty-four blastocysts derived from vitrified-thawed oocytes were transferred to six Luxi yellow cattle recipients. Two recipients (33%) were diagnosed pregnant; one aborted 97 d after transfer, whereas the other delivered a cloned calf after 263 d. As a control, 28 synchronous Luxi yellow cattle recipients each received a single blastocyst produced using a fresh oocyte as a nuclear recipient; 10 recipients were diagnosed pregnant, of which 6 (21.4% of the original 28) delivered cloned calves. In conclusion, bovine oocytes vitrified by the OPS method and subsequently thawed supported development (to term) of somatic cell cloned embryos.

  10. Transforming Elementary Science Teacher Education by Bridging Formal and Informal Science Education in an Innovative Science Methods Course

    ERIC Educational Resources Information Center

    Riedinger, Kelly; Marbach-Ad, Gili; McGinnis, J. Randy; Hestness, Emily; Pease, Rebecca

    2011-01-01

    We investigated curricular and pedagogical innovations in an undergraduate science methods course for elementary education majors at the University of Maryland. The goals of the innovative elementary science methods course included: improving students' attitudes toward and views of science and science teaching, to model innovative science teaching…

  11. Innovation, Innovation, Innovation

    ERIC Educational Resources Information Center

    Schuller, Tom

    2007-01-01

    Innovation, Universities and Skills. The new title of the department offers much food for thought. The title is indeed an intriguing and important one. Bringing the idea of innovation right to the fore is, to use an overworked term, challenging. Pinning down what innovation means is not at all easy. There are three different lines of argument. The…

  12. Comprehensive census of bacteria in clean rooms by using DNA microarray and cloning methods.

    PubMed

    La Duc, Myron T; Osman, Shariff; Vaishampayan, Parag; Piceno, Yvette; Andersen, Gary; Spry, J A; Venkateswaran, Kasthuri

    2009-10-01

    A census of clean room surface-associated bacterial populations was derived from the results of both the cloning and sequencing of 16S rRNA genes and DNA microarray (PhyloChip) analyses. Samples from the Lockheed Martin Aeronautics Multiple Testing Facility (LMA-MTF), the Kennedy Space Center Payload Hazard and Servicing Facility (KSC-PHSF), and the Jet Propulsion Laboratory Spacecraft Assembly Facility (JPL-SAF) clean rooms were collected during the various assembly phases of the Phoenix and Mars Science Laboratory (MSL) spacecraft. Clone library-derived analyses detected a larger bacterial diversity prior to the arrival of spacecraft hardware in these clean room facilities. PhyloChip results were in agreement with this trend but also unveiled the presence of anywhere from 9- to 70-fold more bacterial taxa than cloning approaches. Among the facilities sampled, the JPL-SAF (MSL mission) housed a significantly less diverse bacterial population than either the LMA-MTF or KSC-PHSF (Phoenix mission). Bacterial taxa known to thrive in arid conditions were frequently detected in MSL-associated JPL-SAF samples, whereas proteobacterial lineages dominated Phoenix-associated KSC-PHSF samples. Comprehensive bacterial censuses, such as that reported here, will help space-faring nations preemptively identify contaminant biomatter that may compromise extraterrestrial life detection experiments. The robust nature and high sensitivity of DNA microarray technologies should prove beneficial to a wide range of scientific, electronic, homeland security, medical, and pharmaceutical applications and to any other ventures with a vested interest in monitoring and controlling contamination in exceptionally clean environments. PMID:19700540

  13. Innovative methods in soil phosphorus research: A review

    PubMed Central

    Kruse, Jens; Abraham, Marion; Amelung, Wulf; Baum, Christel; Bol, Roland; Kühn, Oliver; Lewandowski, Hans; Niederberger, Jörg; Oelmann, Yvonne; Rüger, Christopher; Santner, Jakob; Siebers, Meike; Siebers, Nina; Spohn, Marie; Vestergren, Johan; Vogts, Angela; Leinweber, Peter

    2015-01-01

    Phosphorus (P) is an indispensable element for all life on Earth and, during the past decade, concerns about the future of its global supply have stimulated much research on soil P and method development. This review provides an overview of advanced state-of-the-art methods currently used in soil P research. These involve bulk and spatially resolved spectroscopic and spectrometric P speciation methods (1 and 2D NMR, IR, Raman, Q-TOF MS/MS, high resolution-MS, NanoSIMS, XRF, XPS, (µ)XAS) as well as methods for assessing soil P reactions (sorption isotherms, quantum-chemical modeling, microbial biomass P, enzymes activity, DGT, 33P isotopic exchange, 18O isotope ratios). Required experimental set-ups and the potentials and limitations of individual methods present a guide for the selection of most suitable methods or combinations. PMID:26167132

  14. Repeatable construction method for engineered zinc finger nuclease based on overlap extension PCR and TA-cloning.

    PubMed

    Fujii, Wataru; Kano, Kiyoshi; Sugiura, Koji; Naito, Kunihiko

    2013-01-01

    Zinc finger nuclease (ZFN) is a useful tool for endogenous site-directed genome modification. The development of an easier, less expensive and repeatedly usable construction method for various sequences of ZFNs should contribute to the further widespread use of this technology. Here, we establish a novel construction method for ZFNs. Zinc finger (ZF) fragments were synthesized by PCR using short primers coding DNA recognition helices of the ZF domain. DNA-binding domains composed of 4 to 6 ZFs were synthesized by overlap extension PCR of these PCR products, and the DNA-binding domains were joined with a nuclease vector by TA cloning. The short primers coding unique DNA recognition helices can be used repeatedly for other ZFN constructions. By using this novel OLTA (OverLap extension PCR and TA-cloning) method, arbitrary ZFN vectors were synthesized within 3 days, from the designing to the sequencing of the vector. Four different ZFN sets synthesized by OLTA showed nuclease activities at endogenous target loci. Genetically modified mice were successfully generated using ZFN vectors constructed by OLTA. This method, which enables the construction of intended ZFNs repeatedly and inexpensively in a short period of time, should contribute to the advancement of ZFN technology.

  15. INNOVATIVE METHODS FOR THE OPTIMIZATION OF GRAVITY STORM SEWER DESIGN

    EPA Science Inventory

    The purpose of this paper is to describe a new method for optimizing the design of urban storm sewer systems. Previous efforts to optimize gravity sewers have met with limited success because classical optimization methods require that the problem be well behaved, e.g. describ...

  16. FIELD ANALYTICAL SCREENING PROGRAM: PCP METHOD - INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    The Field Analytical Screening Program (FASP) pentachlorophenol (PCP) method uses a gas chromatograph (GC) equipped with a megabore capillary column and flame ionization detector (FID) and electron capture detector (ECD) to identify and quantify PCP. The FASP PCP method is design...

  17. Why Clone?

    MedlinePlus

    ... How might cloning be used in medicine? Cloning animal models of disease Much of what researchers learn about human disease comes from studying animal models such as mice. Often, animal models are ...

  18. What Discourages Students from Engaging with Innovative Instructional Methods: Creating a Barrier Framework

    ERIC Educational Resources Information Center

    Ellis, Donna E.

    2015-01-01

    When faculty members choose to implement instructional methods that are learning-centred, this may represent a change for students; and some resist engaging. In this exploratory case study research, 172 students shared what discourages them from being willing to engage with these innovative methods that aim to facilitate their learning.…

  19. Farmers' Preferences for Methods of Receiving Information on New or Innovative Farming Practices.

    ERIC Educational Resources Information Center

    Riesenberg, Lou E.; Gor, Christopher Obel

    1989-01-01

    Survey of 386 Idaho farmers (response rate 58 percent) identified preferred methods of receiving information on new or innovative farming practices. Analysis revealed preference for interpersonal methods (demonstrations, tours, and field trips) over mass media such as computer-assisted instruction (CAI) and home study, although younger farmers,…

  20. Single-step cloning-screening method: a new tool for developing and studying high-titer viral vector producer cells.

    PubMed

    Rodrigues, A F; Formas-Oliveira, A S; Guerreiro, M R; Tomás, H A; Alves, P M; Coroadinha, A S

    2015-09-01

    This article describes a novel method merging the cloning of viral vector producer cells with vector titer screening, allowing for screening 200-500 clones in 2 weeks. It makes use of a GFP separated into two fragments, S10 and S11 (Split GFP), fluorescing only upon transcomplementation. Producer cells carrying a S11 viral transgene are cloned in 96-well plates and co-cultured with target cells stably expressing S10. During the period of clone expansion, S11 viruses infect S10 target cells reconstituting the GFP signal. Transcomplemented fluorescence data provide direct estimation of the clone's productivity and can be analyzed in terms of density distribution, offering valuable information on the average productivity of the cell population and allowing the identification of high-producing clones. The method was validated by establishing a retrovirus producer from a nude cell line, in <3 months, inserting three vector constructs without clone selection or screening in between. Clones producing up to 10(8) infectious particles per ml were obtained, delivering optimal ratios of infectious-to-total particles (1 to 5). The method was additionally used to evaluate the production performance of HEK 293 and HEK 293T cell lines demonstrating that the latter sustains increased titers. Finally, it was used to study genetic manipulation of glutathione metabolism in retrovirus production showing that changing cell metabolism steers higher vector expression with titer increases of more than one order of magnitude.This method is a valuable tool not only for cell line development but also for genetic manipulation of viral vector and/or producer cells contributing to advancing the field of viral gene therapy.

  1. An Innovative Method to Study Stokes' Law in the Laboratory

    ERIC Educational Resources Information Center

    Wadhwa, Ajay

    2008-01-01

    A new method is introduced to study the behaviour of the falling spherical ball in a viscous liquid using the well known Stokes' law. Experimental results are compared with those obtained by numerical calculations. Upper limits on the size and mass of the spherical balls of different materials used in the experiment are presented. (Contains 5…

  2. Estimation of microbial cover distributions at Mammoth Hot Springs using a multiple clone library resampling method.

    PubMed

    Martín, Héctor García; Goldenfeld, Nigel

    2006-07-01

    We propose the use of cover as a quick, low-resolution proxy for the abundance of microbial species, which reduces polymerase chain reaction bias. We showcase this concept in a computation that uses clone library information from travertine-forming hot springs in Yellowstone National Park to provide estimates of relative covers at different locations within the spring system. Samples were used from two media: the water column and the travertine substrate. The cover distribution is found to approximate a power law for samples within the water column. Significant commonality of species with the highest cover is observed in the water column for all locations, but not for species present in the substrate at different locations or between media at the same location. PMID:16817923

  3. Implementation of optimal phase-covariant cloning machines

    SciTech Connect

    Sciarrino, Fabio; De Martini, Francesco

    2007-07-15

    The optimal phase-covariant quantum cloning machine (PQCM) broadcasts the information associated to an input qubit into a multiqubit system, exploiting a partial a priori knowledge of the input state. This additional a priori information leads to a higher fidelity than for the universal cloning. The present article first analyzes different innovative schemes to implement the 1{yields}3 PQCM. The method is then generalized to any 1{yields}M machine for an odd value of M by a theoretical approach based on the general angular momentum formalism. Finally different experimental schemes based either on linear or nonlinear methods and valid for single photon polarization encoded qubits are discussed.

  4. [Cloning - controversies].

    PubMed

    Twardowski, T; Michalska, A

    2001-01-01

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

  5. The Elnady Technique: An innovative, new method for tissue preservation.

    PubMed

    Elnady, Fawzy A

    2016-01-01

    At the Faculty of Veterinary Medicine, Cairo University, there is an increasing number of students but a limited availability of animal cadavers used for dissection, and student exposure to formalin is a known hazard. In order to address these challenges, a new method for tissue preservation was developed, the "Elnady Technique." This method is a modified form of plastination, where the chemicals used are not patented, are inexpensive and locally available, and the process is performed at room temperature. The produced specimens are realistic, durable, have no offensive odor, and are dry, soft and flexible. They can be used to replace the use of animals killed for teaching basic anatomy, embryology, pathology, parasitology and forensic medicine. They have great potential to support training in clinical skills and surgery, including for clinical examination, endoscopy, surgical sutures, and obstetrics simulation. PMID:26821550

  6. Educational research methods for researching innovations in teaching, learning and assessment: The nursing lecturer as researcher.

    PubMed

    Marks-Maran, Diane

    2015-11-01

    The author, who has had previous experience as a nurse researcher, has been engaged in helping nurse lecturers to undertake evaluation research studies into innovations in their teaching, learning and assessment methods. In order to undertake this work successfully, it was important to move from thinking like a nurse researcher to thinking like an educational researcher and developing the role of the nursing lecturer as researcher of their teaching. This article explores the difference between evaluation and evaluation research and argues for the need to use educational research methods when undertaking evaluation research into innovations in teaching, learning and assessment. A new model for educational evaluation research is presented together with two case examples of the model in use. The model has been tested on over 30 research studies into innovations in teaching, learning and assessment over the past 8 years.

  7. Educational research methods for researching innovations in teaching, learning and assessment: The nursing lecturer as researcher.

    PubMed

    Marks-Maran, Diane

    2015-11-01

    The author, who has had previous experience as a nurse researcher, has been engaged in helping nurse lecturers to undertake evaluation research studies into innovations in their teaching, learning and assessment methods. In order to undertake this work successfully, it was important to move from thinking like a nurse researcher to thinking like an educational researcher and developing the role of the nursing lecturer as researcher of their teaching. This article explores the difference between evaluation and evaluation research and argues for the need to use educational research methods when undertaking evaluation research into innovations in teaching, learning and assessment. A new model for educational evaluation research is presented together with two case examples of the model in use. The model has been tested on over 30 research studies into innovations in teaching, learning and assessment over the past 8 years. PMID:25630227

  8. Innovative research methods for studying treatments for rare diseases: methodological review

    PubMed Central

    Thompson, Lauren; O’Keefe, Kelly; Kesselheim, Aaron S

    2014-01-01

    Objective To examine methods for generating evidence on health outcomes in patients with rare diseases. Design Methodological review of existing literature. Setting PubMed, Embase, and Academic Search Premier searched for articles describing innovative approaches to randomized trial design and analysis methods and methods for conducting observational research in patients with rare diseases. Main outcome measures We assessed information related to the proposed methods, the specific rare disease being studied, and outcomes from the application of the methods. We summarize methods with respect to their advantages in studying health outcomes in rare diseases and provide examples of their application. Results We identified 46 articles that proposed or described methods for studying patient health outcomes in rare diseases. Articles covered a wide range of rare diseases and most (72%) were published in 2008 or later. We identified 16 research strategies for studying rare disease. Innovative clinical trial methods minimize sample size requirements (n=4) and maximize the proportion of patients who receive active treatment (n=2), strategies crucial to studying small populations of patients with limited treatment choices. No studies describing unique methods for conducting observational studies in patients with rare diseases were identified. Conclusions Though numerous studies apply unique clinical trial designs and considerations to assess patient health outcomes in rare diseases, less attention has been paid to innovative methods for studying rare diseases using observational data. PMID:25422272

  9. [Investigation of bacterial diversity in the biological desulfurization reactor for treating high salinity wastewater by the 16S rDNA cloning method].

    PubMed

    Liu, Wei-Guo; Liang, Cun-Zhen; Yang, Jin-Sheng; Wang, Gui-Ping; Liu, Miao-Miao

    2013-02-01

    The bacterial diversity in the biological desulfurization reactor operated continuously for 1 year was studied by the 16S rDNA cloning and sequencing method. Forty clones were randomly selected and their partial 16S rDNA genes (ca. 1,400 bp) were sequenced and blasted. The results indicated that there were dominant bacterias in the biological desulfurization reactor, where 33 clones belonged to 3 different published phyla, while 1 clone belonged to unknown phylum. The dominant bacterial community in the system was Proteobacteria, which accounted for 85.3%. The bacterial community succession was as follows: the gamma-Proteobacteria(55.9%), beta-Proteobacteria(17.6%), Actinobacteridae (8.8%), delta-Proteobacteria (5.9%) , alpha-Proteobacteria(5.9%), and Sphingobacteria (2.9%). Halothiobacillus sp. ST15 and Thiobacillus sp. UAM-I were the major desulfurization strains.

  10. An Innovative Operative Method for Correction of Tracheal Stenosis.

    PubMed

    Shah, Dakshesh R; Mungutwar, Varsha

    2016-09-01

    This is a case of failed attempt of hanging, resulted into cervical injury & injury to the trachea. Patient was put on the ventilator for respiratory support for long period. Poor recovery of the patient resulted into prolong tracheostomy tube. Combine effect of all resulted into suprastomal tracheal stenosis. Due to complexity of the problem a novel method was customized to correct the tracheal stenosis. Strap muscle (Sternohyoid & Sternothyroid) was raised as a flap; free cartilage was sandwiched in between and sutured to the created tracheal defect. Bovine collagen was placed over a stent and placed in the tracheal lumen. Good result was achieved post operatively in the form of adequate lumen, rigid anterior wall during respiration, good mucosal lining without fibrosis and normal speech. PMID:27508127

  11. Innovative methods of knowledge transfer by multimedia library

    NASA Astrophysics Data System (ADS)

    Goanta, A. M.

    2016-08-01

    The present situation of teaching and learning new knowledge taught in the classroom is highly variable depending on the specific topics concerned. If we analyze the manifold ways of teaching / learning at university level, we can notice a very good combination between classical and modern methods. The first category includes the classic chalk blackboard teaching, followed by the also classical learning based on paper reference material. The second category includes books published in PDF or PPT [1], which are printed on the type backing CD / DVD. Since 2006 the author was concerned about the transfer of information and knowledge through video files like AVI, FLV or MPEG using various means of transfer, from the free ones (via Internet) and continuing with those involving minimal costs, i.e. on CD / DVD support. Encouraged by the students’ interest in this kind of teaching material as proved by monitoring [2] the site http://www.cursuriuniversitarebraila.ugal.ro, the author has managed to publish with ISBN the first video book in Romania, which has a non conformist content in that the chapters are located not by paging but by the hour or minutes of shooting when they were made.

  12. New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

    DOE PAGESBeta

    Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; Kulkkarni, Gargi; Metcalf, William W.

    2008-01-01

    A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri P mcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline inmore » strains that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR -regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.« less

  13. A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep.

    PubMed

    Friis, Thor Einar; Stephenson, Sally; Xiao, Yin; Whitehead, Jon; Hutmacher, Dietmar W

    2014-10-01

    The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes.

  14. New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

    PubMed Central

    Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; Kulkkarni, Gargi; Metcalf, William W.

    2008-01-01

    A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri PmcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strains that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR-regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion. PMID:19054746

  15. Key Competencies and Characteristics for Innovative Teaching among Secondary School Teachers: A Mixed-Methods Research

    ERIC Educational Resources Information Center

    Zhu, Chang; Wang, Di

    2014-01-01

    This research aims to understand the key competencies and characteristics for innovative teaching as perceived by Chinese secondary teachers. A mixed-methods research was used to investigate secondary teachers' views. First, a qualitative study was conducted with interviews of teachers to understand the perceived key competencies and…

  16. Building America Top Innovations 2014 Profile: HVAC Cabinet Air Leakage Test Method

    SciTech Connect

    none,

    2014-11-01

    This 2014 Top Innovation profile describes Building America-funded research by teams and national laboratories that resulted in the development of an ASHRAE standard and a standardized testing method for testing the air leakage of HVAC air handlers and furnace cabinets and has spurred equipment manufacturers to tighten the cabinets they use for residential HVAC systems.

  17. Co-Designing and Co-Teaching Graduate Qualitative Methods: An Innovative Ethnographic Workshop Model

    ERIC Educational Resources Information Center

    Cordner, Alissa; Klein, Peter T.; Baiocchi, Gianpaolo

    2012-01-01

    This article describes an innovative collaboration between graduate students and a faculty member to co-design and co-teach a graduate-level workshop-style qualitative methods course. The goal of co-designing and co-teaching the course was to involve advanced graduate students in all aspects of designing a syllabus and leading class discussions in…

  18. Modeling technology innovation: How science, engineering, and industry methods can combine to generate beneficial socioeconomic impacts

    PubMed Central

    2012-01-01

    Background Government-sponsored science, technology, and innovation (STI) programs support the socioeconomic aspects of public policies, in addition to expanding the knowledge base. For example, beneficial healthcare services and devices are expected to result from investments in research and development (R&D) programs, which assume a causal link to commercial innovation. Such programs are increasingly held accountable for evidence of impact—that is, innovative goods and services resulting from R&D activity. However, the absence of comprehensive models and metrics skews evidence gathering toward bibliometrics about research outputs (published discoveries), with less focus on transfer metrics about development outputs (patented prototypes) and almost none on econometrics related to production outputs (commercial innovations). This disparity is particularly problematic for the expressed intent of such programs, as most measurable socioeconomic benefits result from the last category of outputs. Methods This paper proposes a conceptual framework integrating all three knowledge-generating methods into a logic model, useful for planning, obtaining, and measuring the intended beneficial impacts through the implementation of knowledge in practice. Additionally, the integration of the Context-Input-Process-Product (CIPP) model of evaluation proactively builds relevance into STI policies and programs while sustaining rigor. Results The resulting logic model framework explicitly traces the progress of knowledge from inputs, following it through the three knowledge-generating processes and their respective knowledge outputs (discovery, invention, innovation), as it generates the intended socio-beneficial impacts. It is a hybrid model for generating technology-based innovations, where best practices in new product development merge with a widely accepted knowledge-translation approach. Given the emphasis on evidence-based practice in the medical and health fields and

  19. Comparative analysis of eukaryotic marine microbial assemblages from 18S rRNA gene and gene transcript clone libraries by using different methods of extraction.

    PubMed

    Koid, Amy; Nelson, William C; Mraz, Amy; Heidelberg, Karla B

    2012-06-01

    Eukaryotic marine microbes play pivotal roles in biogeochemical nutrient cycling and ecosystem function, but studies that focus on the protistan biogeography and genetic diversity lag-behind studies of other microbes. 18S rRNA PCR amplification and clone library sequencing are commonly used to assess diversity that is culture independent. However, molecular methods are not without potential biases and artifacts. In this study, we compare the community composition of clone libraries generated from the same water sample collected at the San Pedro Ocean Time Series (SPOTs) station in the northwest Pacific Ocean. Community composition was assessed using different cell lysis methods (chemical and mechanical) and the extraction of different nucleic acids (DNA and RNA reverse transcribed to cDNA) to build Sanger ABI clone libraries. We describe specific biases for ecologically important phylogenetic groups resulting from differences in nucleic acid extraction methods that will inform future designs of eukaryotic diversity studies, regardless of the target sequencing platform planned.

  20. Academic Cloning.

    ERIC Educational Resources Information Center

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

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

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

    PubMed Central

    Throop, Andrea L.; LaBaer, Joshua

    2015-01-01

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

  2. Plant Methods: putting the spotlight on technological innovation in the plant sciences.

    PubMed

    Forde, Brian G; Roberts, Michael R

    2005-01-01

    Plant Methods is a new journal for plant biologists, specialising in the rapid publication of peer-reviewed articles with a focus on technological innovation in the plant sciences. The aim of Plant Methods is to stimulate the development and adoption of new and improved techniques and research tools in plant biology. We hope to promote more consistent standards in the plant sciences, and make readily accessible laboratory and computer-based research tools available to the whole community. This will be achieved by publishing Research articles, Methodology papers and Reviews using the BioMed Central Open Access publishing model. The journal is supported by a prestigious editorial board, whose members all recognise the importance of technological innovation as a driver for basic science.

  3. Innovations in teaching/learning methods for medical students: research with mentoring.

    PubMed

    Kotwal, Atul

    2013-01-01

    In this era of evidence-based medicine, innovative teaching learning methods coupled with appropriate mentoring are required to make the undergraduate medical students interested in the speciality of Community Medicine. Teaching research methodology to medical students by a hands-on approach along with mentoring by involving them in small, feasible, sustainable, and replicable studies on a topic of interest clearly showed renewed interest among students toward the subject. PMID:24125928

  4. High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe.

    PubMed Central

    Okazaki, K; Okazaki, N; Kume, K; Jinno, S; Tanaka, K; Okayama, H

    1990-01-01

    We describe a highly efficient alkali cation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of fission yeast Schizosaccharomyces pombe mutants. cDNA libraries constructed with the pcD or pcD2 vector are transduced into yeast by cotransfection with a linearized vector, which allows an enhanced homologous recombination between the yeast vector and the library plasmid leading to the efficient formation of concatemers containing pcD molecules. The transformation frequencies obtained by the method are 10(6) colonies per 10(8) cells transfected with 2 micrograms of library and 1 microgram of vector, 50-60% of which contain pcD molecules. The high-efficiency alkali cation method circumvents many of the shortcomings of the spheroplast method generally used for Schiz. pombe transfection. The vectors are maximized for the efficiency of library transduction and minimized for the rearrangements of pcD molecules during propagation in yeast. This system allows rapid screening of multi-million cDNA clone libraries for rare cDNAs in a routine scale of experiments. Using this system, various mammalian cDNAs that are extremely difficult, time-consuming, or unclonable to clone by other methods have been cloned. Images PMID:2251111

  5. Methods of plate pexophagy monitoring and positive selection for ATG gene cloning in yeasts.

    PubMed

    Stasyk, Oleh V; Nazarko, Taras Y; Sibirny, Andriy A

    2008-01-01

    Methods for colony assay of peroxisomal oxidases in yeasts provide a convenient and fast approach for monitoring peroxisome status. They have been used in several laboratories for the isolation of yeast mutants deficient in selective autophagic peroxisome degradation (pexophagy), catabolite repression of peroxisomal enzymes or mutants deficient in oxidases themselves. In this chapter, protocols for monitoring peroxisomal alcohol oxidase and amine oxidase directly in yeast colonies and examples of their application for mutant isolation are described. These methods were successfully utilized in several methylotrophic yeasts and the alkane-utilizing yeast Yarrowia lipolytica.

  6. Molecular Profiling of Microbial Communities from Contaminated Sources: Use of Subtractive Cloning Methods and rDNA Spacer Sequences

    SciTech Connect

    Robb, Frank T.

    2001-04-10

    The major objective of this research was to provide appropriate sequences and assemble a DNA array of oligonucleotides to be used for rapid profiling of microbial populations from polluted areas and other areas of interest. The sequences to be assigned to the DNA array were chosen from cloned genomic DNA taken from groundwater sites having well characterized pollutant histories at Hanford Nuclear Plant and Lawrence Livermore Site 300. Glass-slide arrays were made and tested; and a new multiplexed, bead-based method was developed that uses nucleic acid hybridization on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences. The test data revealed considerable strain variation between sample sites showing a striking distribution of sequences. It also suggests that diversity varies greatly with bioremediation, and that there are many bacterial intergenic spacer region sequences that can indicate its effects. The bead method exhibited superior sequence discrimination and has features for easier and more accurate measurement.

  7. A PCR-based method for manipulation of the vaccinia virus genome that eliminates the need for cloning.

    PubMed

    Turner, P C; Moyer, R W

    1992-11-01

    A general method is described for altering specific genes of vaccinia virus (VV). We demonstrate and evaluate the procedure by gene inactivation, using a dominant selectable marker in conjunction with recombinant polymerase chain reaction (PCR). Primers based on the sequence of the target gene enable amplification of flanking arms and their subsequent attachment to the gpt cassette that confers resistance to mycophenolic acid. Linear PCR constructs are transfected into cells infected with wild-type vaccinia virus. Mutant viruses with gpt inserted into the target gene by homologous recombination are then selected by growth in the presence of MPA. This technique was applied to the vaccinia virus thymidine kinase gene and compared to the traditional method of constructing gpt-containing plasmids by cloning. The PCR scheme was found to be highly efficient and could theoretically be used to insert any foreign DNA element into any nonessential target gene for which partial or complete sequence information is available. The procedure can potentially be used for a wide variety of genetic modifications, including the insertion of foreign genes, with poxviruses and other DNA viruses. Genomes of microorganisms, such as bacteria and yeast that can be transformed with linear DNA, are also candidates for manipulation by this methodology.

  8. A Novel Cloning Template Designing Method by Using an Artificial Bee Colony Algorithm for Edge Detection of CNN Based Imaging Sensors

    PubMed Central

    Parmaksızoğlu, Selami; Alçı, Mustafa

    2011-01-01

    Cellular Neural Networks (CNNs) have been widely used recently in applications such as edge detection, noise reduction and object detection, which are among the main computer imaging processes. They can also be realized as hardware based imaging sensors. The fact that hardware CNN models produce robust and effective results has attracted the attention of researchers using these structures within image sensors. Realization of desired CNN behavior such as edge detection can be achieved by correctly setting a cloning template without changing the structure of the CNN. To achieve different behaviors effectively, designing a cloning template is one of the most important research topics in this field. In this study, the edge detecting process that is used as a preliminary process for segmentation, identification and coding applications is conducted by using CNN structures. In order to design the cloning template of goal-oriented CNN architecture, an Artificial Bee Colony (ABC) algorithm which is inspired from the foraging behavior of honeybees is used and the performance analysis of ABC for this application is examined with multiple runs. The CNN template generated by the ABC algorithm is tested by using artificial and real test images. The results are subjectively and quantitatively compared with well-known classical edge detection methods, and other CNN based edge detector cloning templates available in the imaging literature. The results show that the proposed method is more successful than other methods. PMID:22163903

  9. A novel multigene cloning method for the production of a motile ATPase.

    PubMed

    Jang, Min Su; Song, Woo Chul; Shin, Seung Won; Park, Kyung Soo; Kim, Jinseok; Kim, Dong-Ik; Kim, Byung Woo; Um, Soong Ho

    2015-08-10

    With the advent of nanotechnology, new functional modules (e.g., nanomotors, nanoprobes) have become essential in several medical fields. Generally, mechanical modulators systems are the principal components of most cutting-edge technologies in modern biomedical applications. However, the in vivo use of motile probes has raised many concerns due to their low sensitivity and non-biocompatibility. As an alternative, biological enzymatic engines have received increased attention. In particular, ATPases, which belong to a class of motile enzymes that catalyze chemical metabolic reactions, have emerged as a promising motor due to their improved biocompatibility and performance. However, ATPases usually suffer from lower functional activity and are difficult to express recombinantly in bacteria relative to their conventional and synthetic competitors. Here, we report a novel functional modified ATPase with both a simple purification protocol and enhanced motile activity. For this mutant ATPase, a new bacterial subcloning method was established. The ATPase-encoding sequence was redesigned so that the mutant ATPase could be easily produced in an Escherichia coli system. The modified thermophilic F1-ATPase (mTF1-ATPase) demonstrated 17.8unit/mg ATPase activity. We propose that derivatives of our ATPase may enable the development of novel in vitro and in vivo synthetic medical diagnostics, as well as therapeutics.

  10. Innovative Methods of Male Circumcision for HIV Prevention—Getting the Right Evidence

    PubMed Central

    Hargreave, Timothy; Ridzon, Renee; Farley, Tim

    2016-01-01

    Abstract: World Health Organization recommends that countries with hyperendemic and generalized HIV epidemics implement voluntary medical male circumcision programs for HIV prevention. Innovative methods of male circumcision including devices have the potential to simplify the procedure, reduce time and cost, increase client acceptability, enhance safety, and expand the numbers of providers who may perform circumcision. We describe work led by World Health Organization and supported by global partners to define a pathway for the evaluation of efficacy and safety of male circumcision devices, to set priority criteria, and to establish a process to guide the use of devices in publicly funded voluntary medical male circumcision programs for HIV prevention. A device classification scheme, an expert Technical Advisory Group on Innovations in Male Circumcision, and a formal prequalification program have also guided considerations on safe use of devices. A rigorous approach was deemed appropriate given the intervention is for use among healthy men for public health purposes. The pathway and processes led to coordinated research, better standardization in research outcomes, and guidance that informed the research, introduction and implementation phases. The lessons learnt from this case study can inform evaluation and use of future public health innovations. PMID:27331591

  11. Innovative Methods of Male Circumcision for HIV Prevention-Getting the Right Evidence.

    PubMed

    Samuelson, Julia; Hargreave, Timothy; Ridzon, Renee; Farley, Tim

    2016-06-01

    World Health Organization recommends that countries with hyperendemic and generalized HIV epidemics implement voluntary medical male circumcision programs for HIV prevention. Innovative methods of male circumcision including devices have the potential to simplify the procedure, reduce time and cost, increase client acceptability, enhance safety, and expand the numbers of providers who may perform circumcision. We describe work led by World Health Organization and supported by global partners to define a pathway for the evaluation of efficacy and safety of male circumcision devices, to set priority criteria, and to establish a process to guide the use of devices in publicly funded voluntary medical male circumcision programs for HIV prevention. A device classification scheme, an expert Technical Advisory Group on Innovations in Male Circumcision, and a formal prequalification program have also guided considerations on safe use of devices. A rigorous approach was deemed appropriate given the intervention is for use among healthy men for public health purposes. The pathway and processes led to coordinated research, better standardization in research outcomes, and guidance that informed the research, introduction and implementation phases. The lessons learnt from this case study can inform evaluation and use of future public health innovations.

  12. Innovative Methods of Male Circumcision for HIV Prevention-Getting the Right Evidence.

    PubMed

    Samuelson, Julia; Hargreave, Timothy; Ridzon, Renee; Farley, Tim

    2016-06-01

    World Health Organization recommends that countries with hyperendemic and generalized HIV epidemics implement voluntary medical male circumcision programs for HIV prevention. Innovative methods of male circumcision including devices have the potential to simplify the procedure, reduce time and cost, increase client acceptability, enhance safety, and expand the numbers of providers who may perform circumcision. We describe work led by World Health Organization and supported by global partners to define a pathway for the evaluation of efficacy and safety of male circumcision devices, to set priority criteria, and to establish a process to guide the use of devices in publicly funded voluntary medical male circumcision programs for HIV prevention. A device classification scheme, an expert Technical Advisory Group on Innovations in Male Circumcision, and a formal prequalification program have also guided considerations on safe use of devices. A rigorous approach was deemed appropriate given the intervention is for use among healthy men for public health purposes. The pathway and processes led to coordinated research, better standardization in research outcomes, and guidance that informed the research, introduction and implementation phases. The lessons learnt from this case study can inform evaluation and use of future public health innovations. PMID:27331591

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

    PubMed Central

    Lipschutz, J H

    1999-01-01

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

  14. An Innovative Method for Dynamic Characterization of Fan FilterUnit Operation.

    SciTech Connect

    Xu, Tengfang

    2006-12-21

    Fan filter units (FFU) are widely used to deliver re-circulated air while providing filtration control of particle concentration in controlled environments such as cleanrooms, minienvironments, and operating rooms in hospitals. The objective of this paper is to document an innovative method for characterizing operation and control of an individual fan filter unit within its operable conditions. Built upon the draft laboratory method previously published [1] , this paper presents an updated method including a testing procedure to characterize dynamic operation of fan filter units, i.e., steady-state operation conditions determined by varied control schemes, airflow rates, and pressure differential across the units. The parameters for dynamic characterization include total electric power demand, total pressure efficiency, airflow rate, pressure differential across fan filter units, and airflow uniformity.

  15. Innovative Method Using Magnetic Particle Tracking to Measure Solids Circulation in a Spouted Bed

    SciTech Connect

    Patterson, Ms. Emily; Halow, John; Daw, C Stuart

    2010-01-01

    We describe an innovative method for measuring particle motion inside spouted fluidized beds. The method uses a magnetic tracer particle, which follows the bulk particle flow and is continuously tracked by multiple magnetic field detectors located outside the bed. We analyze signals from the detectors to determine the tracer position at each instant in time. From statistical analysis of the tracer trajectory, characteristic measures of the bulk particle flow, such as the average recirculation frequency, can be determined as a function of operating conditions. For experiments with a range of particle sizes and densities in a 3.9-cm-diameter spouted bed, we find that average solids recirculation rates correlate with excess velocity (superficial minus minimum spouting velocity), particle density, and bed depth.

  16. Estimating the Impacts of Local Policy Innovation: The Synthetic Control Method Applied to Tropical Deforestation

    PubMed Central

    Sills, Erin O.; Herrera, Diego; Kirkpatrick, A. Justin; Brandão, Amintas; Dickson, Rebecca; Hall, Simon; Pattanayak, Subhrendu; Shoch, David; Vedoveto, Mariana; Young, Luisa; Pfaff, Alexander

    2015-01-01

    Quasi-experimental methods increasingly are used to evaluate the impacts of conservation interventions by generating credible estimates of counterfactual baselines. These methods generally require large samples for statistical comparisons, presenting a challenge for evaluating innovative policies implemented within a few pioneering jurisdictions. Single jurisdictions often are studied using comparative methods, which rely on analysts’ selection of best case comparisons. The synthetic control method (SCM) offers one systematic and transparent way to select cases for comparison, from a sizeable pool, by focusing upon similarity in outcomes before the intervention. We explain SCM, then apply it to one local initiative to limit deforestation in the Brazilian Amazon. The municipality of Paragominas launched a multi-pronged local initiative in 2008 to maintain low deforestation while restoring economic production. This was a response to having been placed, due to high deforestation, on a federal “blacklist” that increased enforcement of forest regulations and restricted access to credit and output markets. The local initiative included mapping and monitoring of rural land plus promotion of economic alternatives compatible with low deforestation. The key motivation for the program may have been to reduce the costs of blacklisting. However its stated purpose was to limit deforestation, and thus we apply SCM to estimate what deforestation would have been in a (counterfactual) scenario of no local initiative. We obtain a plausible estimate, in that deforestation patterns before the intervention were similar in Paragominas and the synthetic control, which suggests that after several years, the initiative did lower deforestation (significantly below the synthetic control in 2012). This demonstrates that SCM can yield helpful land-use counterfactuals for single units, with opportunities to integrate local and expert knowledge and to test innovations and permutations on

  17. Estimating the Impacts of Local Policy Innovation: The Synthetic Control Method Applied to Tropical Deforestation.

    PubMed

    Sills, Erin O; Herrera, Diego; Kirkpatrick, A Justin; Brandão, Amintas; Dickson, Rebecca; Hall, Simon; Pattanayak, Subhrendu; Shoch, David; Vedoveto, Mariana; Young, Luisa; Pfaff, Alexander

    2015-01-01

    Quasi-experimental methods increasingly are used to evaluate the impacts of conservation interventions by generating credible estimates of counterfactual baselines. These methods generally require large samples for statistical comparisons, presenting a challenge for evaluating innovative policies implemented within a few pioneering jurisdictions. Single jurisdictions often are studied using comparative methods, which rely on analysts' selection of best case comparisons. The synthetic control method (SCM) offers one systematic and transparent way to select cases for comparison, from a sizeable pool, by focusing upon similarity in outcomes before the intervention. We explain SCM, then apply it to one local initiative to limit deforestation in the Brazilian Amazon. The municipality of Paragominas launched a multi-pronged local initiative in 2008 to maintain low deforestation while restoring economic production. This was a response to having been placed, due to high deforestation, on a federal "blacklist" that increased enforcement of forest regulations and restricted access to credit and output markets. The local initiative included mapping and monitoring of rural land plus promotion of economic alternatives compatible with low deforestation. The key motivation for the program may have been to reduce the costs of blacklisting. However its stated purpose was to limit deforestation, and thus we apply SCM to estimate what deforestation would have been in a (counterfactual) scenario of no local initiative. We obtain a plausible estimate, in that deforestation patterns before the intervention were similar in Paragominas and the synthetic control, which suggests that after several years, the initiative did lower deforestation (significantly below the synthetic control in 2012). This demonstrates that SCM can yield helpful land-use counterfactuals for single units, with opportunities to integrate local and expert knowledge and to test innovations and permutations on policies

  18. Estimating the Impacts of Local Policy Innovation: The Synthetic Control Method Applied to Tropical Deforestation.

    PubMed

    Sills, Erin O; Herrera, Diego; Kirkpatrick, A Justin; Brandão, Amintas; Dickson, Rebecca; Hall, Simon; Pattanayak, Subhrendu; Shoch, David; Vedoveto, Mariana; Young, Luisa; Pfaff, Alexander

    2015-01-01

    Quasi-experimental methods increasingly are used to evaluate the impacts of conservation interventions by generating credible estimates of counterfactual baselines. These methods generally require large samples for statistical comparisons, presenting a challenge for evaluating innovative policies implemented within a few pioneering jurisdictions. Single jurisdictions often are studied using comparative methods, which rely on analysts' selection of best case comparisons. The synthetic control method (SCM) offers one systematic and transparent way to select cases for comparison, from a sizeable pool, by focusing upon similarity in outcomes before the intervention. We explain SCM, then apply it to one local initiative to limit deforestation in the Brazilian Amazon. The municipality of Paragominas launched a multi-pronged local initiative in 2008 to maintain low deforestation while restoring economic production. This was a response to having been placed, due to high deforestation, on a federal "blacklist" that increased enforcement of forest regulations and restricted access to credit and output markets. The local initiative included mapping and monitoring of rural land plus promotion of economic alternatives compatible with low deforestation. The key motivation for the program may have been to reduce the costs of blacklisting. However its stated purpose was to limit deforestation, and thus we apply SCM to estimate what deforestation would have been in a (counterfactual) scenario of no local initiative. We obtain a plausible estimate, in that deforestation patterns before the intervention were similar in Paragominas and the synthetic control, which suggests that after several years, the initiative did lower deforestation (significantly below the synthetic control in 2012). This demonstrates that SCM can yield helpful land-use counterfactuals for single units, with opportunities to integrate local and expert knowledge and to test innovations and permutations on policies

  19. Application of Innovative Methods to Optimize the Learning Process in Physics for Medical Students

    NASA Astrophysics Data System (ADS)

    Zlateva, Genoveva; Tsankova, Emilya

    2010-01-01

    Unlike chemistry and biology courses in the high schools which occupy the attention and interest of students as they need to achieve maximum results of examinations for admission in higher medical schools, physics remains away from their interest. Striving for awakening the interest of medical students to classes in physics and diversification of the learning process requires the continuous search of new forms of organization of this process in order to fulfill the main task of education: optimal development of each student, creating conditions for creative work with the highest possible productivity. Using innovations in teaching physics, aimed at the purpose of training in non-traditional way, transforms the passive learning in an active creative process. This allows rapid identification and compensation of gaps in the knowledge, which in turn leads to a rationalization and a more complete and lasting control of educational content. The aim of the study is analysis and evaluation of the effectiveness of the implementation of innovative educational methods to increase motivation and the quality of teaching physics to students of medicine. The discussion is based on the opinions expressed in surveys of students and results of various forms of feedback.

  20. [Students of today, physicians of tomorrow. Educational priorities for innovative teaching methods in medicine].

    PubMed

    Guicciardi, Stefano; Riforgiato, Chiara; Leopardi, Eleonora

    2015-02-01

    In the past three years there has been increasing debate regarding the educational system of physicians in Italy, mainly due to the implementation of radical changes in the selection and admission criteria to medical courses. From the students' point of view, it is possible to identify different educational challenges to be faced in order to develop a more adequate learning system.First of all, it is necessary to elaborate an accurate planning of the physicians based on the needs of the Italian Healthcare System with a proportionate regulation of access to medical and specialty courses. Secondly, there is the strong need to update the traditional curricula based on a biological reductionist perspective with innovative and integrated courses that could teach new essential topics in a radically changed and globalized world, such as global health or medical ethics. Thirdly, it is crucial to introduce innovative teaching methods that could help students improve and evaluate their practical skills and participate actively in their own education, developing the attitudes and competences needed in their future profession. PMID:25734594

  1. An Innovative, Effective and Cost Effective Survey Method Using a Survey-Check Response Format

    PubMed Central

    Feil, Edward G.; Severson, Herbert; Taylor, Ted; Boles, Shawn; Albert, David A.; Blair, Jason

    2007-01-01

    Maximizing the response rate to surveys involves thoughtful choices about survey design, sampling and collection methods. This paper describes an innovative survey method, to provide immediate reinforcement for responding and to minimize the response cost. This method involves using a questionnaire printed as checks on security (anti-fraud) paper with questions and responses separated using a perforated tear off section. Once a participant completes the survey, the response area is detached from the questions, thus protecting the confidentiality of the subject, and the check is returned via the banking system. This report describes the survey-check methodology, the survey flow process, and the results from four research studies which have used this method. These studies include (1) a technology accessibility survey of parents with children enrolled in a low-income preschool program; (2) a parent report of their child’s behavior used as screening criteria for inclusion in a computer-mediated parent education project; (3) a follow-up questionnaire as part of a longitudinal study of child behavior, covering home and classroom interventions, and service utilization, and; (4) a survey of dentists in support of efforts to recruit them to participate in a randomized control trial of tobacco cessation in dental offices. The results of using this method show great improvement in response rates over traditionally administered surveys for three of the four reported studies. Results are discussed in terms of future applications of this method, limitations, and potential cost savings. PMID:17180473

  2. Digestion-ligation-amplification (DLA): a simple genome walking method to amplify unknown sequences flanking mutator (Mu) transposons and thereby facilitate gene cloning.

    PubMed

    Liu, Sanzhen; Hsia, An-Ping; Schnable, Patrick S

    2013-01-01

    Digestion-ligation-amplification (DLA), a novel PCR-based genome walking method, was developed to amplify unknown sequences flanking known sequences of interest. DLA specifically overcomes the problems associated with amplifying genomic sequences flanking high copy number transposons in large genomes. Two DLA-based strategies, MuClone and DLA-454, were developed to isolate Mu-tagged alleles. MuClone allows for the amplification of DNA flanking subsets of the numerous Mu transposons in the genome using unique three-nucleotide tags at the 3'-ends of primers, simplifying the identification of flanking sequences that co-segregate with mutant phenotypes caused by Mu insertions. DLA-454, which combines DLA with 454 pyrosequencing, permits the efficient amplification and sequencing of Mu flanking regions in a high-throughput manner.

  3. An innovative method and experiment for fabricating bulgy shape nanochannel using AFM

    NASA Astrophysics Data System (ADS)

    Lin, Zone-Ching; Jheng, Hao-Yuan; Ding, Hao-Yang

    2015-08-01

    The paper proposes using atomic force microscopy (AFM) and the concept of specific down force energy (SDFE) to establish an innovative offset cycle cutting method for fabricating a bulgy shape nanochannel on a single-crystal silicon substrate. In the offset cycle cutting method, cutting is performed at a constant down force in all cutting passes. After the first cutting pass, the AFM probe is offset rightward for the second pass and subsequently offset leftward to the middle (i.e., between the positions of the first two cutting passes) for the third cutting pass. Applying a step-by-step method to modify the offset distance and approach the defined SDFE value, this study determined the depth of the middle cutting pass and smaller values of upward bulginess and downward indentation at the bottom of the nanochannel. The nanochannel width can be increased by increasing the number of offset cycle cutting passes. In addition, by applying the proposed method, this study involved a simulation and experiment concerning the cutting path plan of bulgy shape nanochannels. Furthermore, using a small down force along the burr path is proposed for reducing burr height. The results of the simulation and experiment were compared to verify the feasibility of the method.

  4. Bioelectrochemical removal of carbon dioxide (CO2): an innovative method for biogas upgrading.

    PubMed

    Xu, Heng; Wang, Kaijun; Holmes, Dawn E

    2014-12-01

    Innovative methods for biogas upgrading based on biological/in-situ concepts have started to arouse considerable interest. Bioelectrochemical removal of CO2 for biogas upgrading was proposed here and demonstrated in both batch and continuous experiments. The in-situ biogas upgrading system seemed to perform better than the ex-situ one, but CO2 content was kept below 10% in both systems. The in-situ system's performance was further enhanced under continuous operation. Hydrogenotrophic methanogenesis and alkali production with CO2 absorption could be major contributors to biogas upgrading. Molecular studies showed that all the biocathodes associated with biogas upgrading were dominated by sequences most similar to the same hydrogenotrophic methanogen species, Methanobacterium petrolearium (97-99% sequence identity). Conclusively, bioelectrochemical removal of CO2 showed great potential for biogas upgrading.

  5. Evaluation of an Innovative Method for Calculating Energy Intake of Hospitalized Patients

    PubMed Central

    Cox Sullivan, Sheila; Bopp, Melinda M.; Roberson, Paula K.; Lensing, Shelly; Sullivan, Dennis H.

    2016-01-01

    The purpose of this study was to evaluate a multi-component method for capturing nutrient intake, which used observation, photography, and an innovative computer program. To assess reliability and accuracy, multiple responsible employees (REs) independently conducted nutrient intake assessments on simulated meals; each RE’s results relating to energy intake were compared to those from the other REs and to those obtained by pre- and post-meal weighing of the food items. System efficiency was assessed by having REs perform independent assessments on the same set of simulated meals using either the new or traditional hospital method for which the REs had to document each food item served and then find the items in a computer database–steps that were automated in the new method. Interrater reliability for energy intake estimated on clinic wards was excellent (intraclass correlation coefficient = 0.975, 95% CI 0.958 to 0.992) and there was a high level of agreement between the REs’ estimates and the true values determined by food weighing; per the method of Bland and Altman the mean difference between the two types of estimates was 0.3 kcal (95% CI, −8.1 to 8.7 kcal) with limits of agreement of −79.5 kcal to 80.1 kcal. Compared to the traditional method, energy intake assessments could be completed using the multi-component method in less than a third of the time. These results indicate the multi-component method is an accurate, reliable, and efficient method of obtaining energy intake assessments for hospitalized patients. PMID:27618096

  6. Evaluation of an Innovative Method for Calculating Energy Intake of Hospitalized Patients.

    PubMed

    Cox Sullivan, Sheila; Bopp, Melinda M; Roberson, Paula K; Lensing, Shelly; Sullivan, Dennis H

    2016-01-01

    The purpose of this study was to evaluate a multi-component method for capturing nutrient intake, which used observation, photography, and an innovative computer program. To assess reliability and accuracy, multiple responsible employees (REs) independently conducted nutrient intake assessments on simulated meals; each RE's results relating to energy intake were compared to those from the other REs and to those obtained by pre- and post-meal weighing of the food items. System efficiency was assessed by having REs perform independent assessments on the same set of simulated meals using either the new or traditional hospital method for which the REs had to document each food item served and then find the items in a computer database-steps that were automated in the new method. Interrater reliability for energy intake estimated on clinic wards was excellent (intraclass correlation coefficient = 0.975, 95% CI 0.958 to 0.992) and there was a high level of agreement between the REs' estimates and the true values determined by food weighing; per the method of Bland and Altman the mean difference between the two types of estimates was 0.3 kcal (95% CI, -8.1 to 8.7 kcal) with limits of agreement of -79.5 kcal to 80.1 kcal. Compared to the traditional method, energy intake assessments could be completed using the multi-component method in less than a third of the time. These results indicate the multi-component method is an accurate, reliable, and efficient method of obtaining energy intake assessments for hospitalized patients. PMID:27618096

  7. Validation of two innovative methods to measure contaminant mass flux in groundwater.

    PubMed

    Goltz, Mark N; Close, Murray E; Yoon, Hyouk; Huang, Junqi; Flintoft, Mark J; Kim, Sehjong; Enfield, Carl

    2009-04-15

    The ability to quantify the mass flux of a groundwater contaminant that is leaching from a source area is critical to enable us to: (1) evaluate the risk posed by the contamination source and prioritize cleanup, (2) evaluate the effectiveness of source remediation technologies or natural attenuation processes, and (3) quantify a source term for use in models that may be applied to predict maximum contaminant concentrations in downstream wells. Recently, a number of new methods have been developed and subsequently applied to measure contaminant mass flux in groundwater in the field. However, none of these methods has been validated at larger than the laboratory-scale through a comparison of measured mass flux and a known flux that has been introduced into flowing groundwater. A couple of innovative flux measurement methods, the tandem circulation well (TCW) and modified integral pumping test (MIPT) methods, have recently been proposed. The TCW method can measure mass flux integrated over a large subsurface volume without extracting water. The TCW method may be implemented using two different techniques. One technique, the multi-dipole technique, is relatively simple and inexpensive, only requiring measurement of heads, while the second technique requires conducting a tracer test. The MIPT method is an easily implemented method of obtaining volume-integrated flux measurements. In the current study, flux measurements obtained using these two methods are compared with known mass fluxes in a three-dimensional, artificial aquifer. Experiments in the artificial aquifer show that the TCW multi-dipole and tracer test techniques accurately estimated flux, within 2% and 16%, respectively; although the good results obtained using the multi-dipole technique may be fortuitous. The MIPT method was not as accurate as the TCW method, underestimating flux by as much as 70%. MIPT method inaccuracies may be due to the fact that the method assumptions (two-dimensional steady

  8. New PCR-based open reading frame typing method for easy, rapid, and reliable identification of Acinetobacter baumannii international epidemic clones without performing multilocus sequence typing.

    PubMed

    Suzuki, Masahiro; Hosoba, Eriko; Matsui, Mari; Arakawa, Yoshichika

    2014-08-01

    Antimicrobial resistance issues have become a global health concern. The rapid identification of multidrug-resistant microbes, which depends on microbial genomic information, is essential for overcoming growing antimicrobial resistance challenges. However, genotyping methods, such as multilocus sequence typing (MLST), for identifying international epidemic clones of Acinetobacter baumannii are not easily performed as routine tests in ordinary clinical laboratories. In this study, we aimed to develop a novel genotyping method that can be performed in ordinary microbiology laboratories. Several open reading frames (ORFs) specific to certain bacterial genetic lineages or species, together with their unique distribution patterns on the chromosomes showing a good correlation with the results of MLST, were selected in A. baumannii and other Acinetobacter spp. by comparing their genomic data. The distribution patterns of the ORFs were visualized by agarose gel electrophoresis after multiplex PCR amplification and digitized. A. baumannii sequence types (STs) corresponding to international clones I and II were successfully discriminated from other STs and Acinetobacter species by detecting the distribution patterns of their ORFs using the multiplex PCR developed here. Since bacterial STs can be easily expressed as digitized numeric data with plus (+) expressed as 1 and minus (-) expressed as 0, the results of the method can be easily compared with those obtained by different tests or laboratories. This PCR-based ORF typing (POT) method can easily and rapidly identify international epidemic clones of A. baumannii and differentiate this microbe from other Acinetobacter spp. Since this POT method is easy enough to be performed even in ordinary clinical laboratories, it would also contribute to daily infection control measures and surveillance.

  9. An innovative high accuracy autonomous navigation method for the Mars rovers

    NASA Astrophysics Data System (ADS)

    Guan, Xujun; Wang, Xinlong; Fang, Jiancheng; Feng, Shaojun

    2014-11-01

    Autonomous navigation is an important function for a Mars rover to fulfill missions successfully. It is a critical technique to overcome the limitations of ground tracking and control traditionally used. This paper proposes an innovative method based on SINS (Strapdown Inertial Navigation System) with the aid of star sensors to accurately determine the rover's position and attitude. This method consists of two parts: the initial alignment and navigation. The alignment consists of a coarse position and attitude initial alignment approach and fine initial alignment approach. The coarse one is used to determine approximate position and attitude for the rover. This is followed by fine alignment to tune the approximate solution to accurate one. Upon the completion of initial alignment, the system can be used to provide real-time navigation solutions for the rover. An autonomous navigation algorithm is proposed to estimate and compensate the accumulated errors of SINS in real time. High accuracy attitude information from star sensor is used to correct errors in SINS. Simulation results demonstrate that the proposed methods can achieve a high precision autonomous navigation for Mars rovers.

  10. [Scientific ethics of human cloning].

    PubMed

    Valenzuela, Carlos Y

    2005-01-01

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

  11. Sequential cloning of chromosomes

    SciTech Connect

    Lacks, S.A.

    1991-12-31

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

  12. Comments on "Comparison of Mann-Kendall and innovative trend method for water quality parameters of the Kizilirmak River, Turkey (Kisi and Ay, 2014)" and "An innovative method for trend analysis of monthly pan evaporations (Kisi, 2015)"

    NASA Astrophysics Data System (ADS)

    Güçlü, Yavuz Selim

    2016-07-01

    Şen (2012) proposed the innovative trend analysis methodology and gave its fundamentals in a simple way based on logical deductions. Later, this methodology has been applied by many writers to different data as mentioned by Kisi and Ay (2014) and Kisi (2015), who also implemented the same methodology. In one of illustrations in their paper (Kisi and Ay, 2014) there are incorrectly scattered data and this comment includes the correction for the illustration. Additionally, Kisi's (2015) application of innovative trend method, as explained in detail by Şen (2012, 2014, 2015), has invalid confidence limits (bands) and the invalidation is explained in the comment.

  13. Human Cloning: Let's Discuss It.

    ERIC Educational Resources Information Center

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

    1999-01-01

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

  14. Innovative sampling and extraction methods for the determination of nonsteroidal anti-inflammatory drugs in water.

    PubMed

    Tanwar, Shivani; Di Carro, Marina; Magi, Emanuele

    2015-03-15

    Two different innovative approaches were used for the determination of nonsteroidal anti-inflammatory drugs (NSAIDs) in water: stir bar sorptive extraction (SBSE) and passive sampling, followed by electrospray ionization liquid chromatography-tandem mass spectrometry. SBSE was developed by comparing EG-Silicone and PDMS stir bars and optimizing main parameters to attain high preconcentration. Quantitative analysis was carried out by mass spectrometry in negative ionization mode and multiple reaction monitoring. The SBSE-LC-MS/MS method provided satisfactory figures of merit with LOD (7.5-71 ng L(-1)) and LOQ (22.5-213 ng L(-1)). The developed method was successfully applied to real samples collected from river water and wastewater effluents. The obtained results showed the presence of all analytes at trace levels, in a wide range of concentrations. The passive sampling approach was carried out by using Polar Organic Chemical Integrative Sampler (POCIS); samplers were deployed for 15 days in river and tap water, allowing to detect analytes at ultra-trace levels. Time-Weighted Average concentration of NSAIDs in river water was estimated in the range 0.33-0.46 ng L(-1), using the sampling rates previously obtained by means of a simple calibration system.

  15. The development of an innovative music therapy treatment method: trial competency through music.

    PubMed

    Sammons, Andrew

    2014-01-01

    Competence to stand trial is necessary for a defendant in criminal adjudication. Recent estimates indicate that between 50,000 and 60,000 defendants in the United States raise the question of competence each year, with approximately 20 percent found incompetent to stand trial (IST). Most of these defendants are committed to an inpatient facility for competence restoration. Although psychopharmacological intervention is a critical component of restoration, as most defendants are found incompetent because of a psychotic disorder, many other modalities of treatment are used. Traditional treatment methods include the use of standardized testing and psychoeducational group sessions. This article discusses the development of an innovative intervention using music therapy. Music as the catalyst provides a forum in which psychiatric patients are engaged and observed within a structured environment designed to address both their factual and rational knowledge and abilities to assist their attorneys in their defense. Trial competency training through a specific music therapy method called Competency Through Music (CTM) is presented, including examples of how music can be used to educate patients and assess trial competence.

  16. The development of an innovative music therapy treatment method: trial competency through music.

    PubMed

    Sammons, Andrew

    2014-01-01

    Competence to stand trial is necessary for a defendant in criminal adjudication. Recent estimates indicate that between 50,000 and 60,000 defendants in the United States raise the question of competence each year, with approximately 20 percent found incompetent to stand trial (IST). Most of these defendants are committed to an inpatient facility for competence restoration. Although psychopharmacological intervention is a critical component of restoration, as most defendants are found incompetent because of a psychotic disorder, many other modalities of treatment are used. Traditional treatment methods include the use of standardized testing and psychoeducational group sessions. This article discusses the development of an innovative intervention using music therapy. Music as the catalyst provides a forum in which psychiatric patients are engaged and observed within a structured environment designed to address both their factual and rational knowledge and abilities to assist their attorneys in their defense. Trial competency training through a specific music therapy method called Competency Through Music (CTM) is presented, including examples of how music can be used to educate patients and assess trial competence. PMID:25187289

  17. A rapid and efficient method for cloning genes of type II restriction-modification systems by use of a killer plasmid.

    PubMed

    Mruk, Iwona; Kaczorowski, Tadeusz

    2007-07-01

    We present a method for cloning restriction-modification (R-M) systems that is based on the use of a lethal plasmid (pKILLER). The plasmid carries a functional gene for a restriction endonuclease having the same DNA specificity as the R-M system of interest. The first step is the standard preparation of a representative, plasmid-borne genomic library. Then this library is transformed with the killer plasmid. The only surviving bacteria are those which carry the gene specifying a protective DNA methyltransferase. Conceptually, this in vivo selection approach resembles earlier methods in which a plasmid library was selected in vitro by digestion with a suitable restriction endonuclease, but it is much more efficient than those methods. The new method was successfully used to clone two R-M systems, BstZ1II from Bacillus stearothermophilus 14P and Csp231I from Citrobacter sp. strain RFL231, both isospecific to the prototype HindIII R-M system. PMID:17468281

  18. An Assessment of the Impact of Implementing Innovative Teaching Methods on Teaching Loads at Golden West College.

    ERIC Educational Resources Information Center

    Parsons, Gary L.

    This study examines the faculty workload policy of a community college that makes extensive use of non-traditional, innovative teaching methods. To measure workload, a mathematical equation whose sum was expressed as 100% was designed to include five factors: instructional hours, number of preparations, weekly student contact hours (WSCH), outside…

  19. Quality improvement of transgenic cloned bovine embryos using an aggregation method: Effects on cell number, cell ratio, embryo perimeter, mitochondrial distribution, and gene expression profile.

    PubMed

    Bang, J I; Jin, J I; Ghanem, N; Choi, B H; Fakruzzaman, M; Ha, A N; Lee, K L; Uhm, S J; Ko, D H; Koo, B C; Lee, J G; Kong, I K

    2015-09-01

    The production of cloned embryos using conventional methods has extremely low success rates owing to low embryo quality. To improve the quality of cloned bovine embryos expressing enhanced green fluorescent protein (EGFP), we applied an aggregation culture method. The EGFP gene was transfected into bovine fetal fibroblasts using a retroviral vector system. Somatic cell nuclear transfer was performed using these cells, and the resulting embryos were cultured in aggregates or individually. Gene expression was analyzed by a microarray, and differentially expressed genes were validated by quantitative real-time polymerase chain reaction. The total number of cells per blastocyst and the ratio of inner cell mass cells to trophectoderm cells were higher in aggregated transgenic cloned blastocysts (agBL; 368.7 ± 109.6 and 1:4.8, respectively) than in in vitro-fertilized blastocysts (ivfBL; 189.8 ± 65.8 and 1:2.6, respectively) and nonaggregated transgenic cloned blastocysts (sBL; 113.1 ± 36.3 and 1:1.5, respectively; P < 0.05 and P < 0.01, respectively). Moreover, the blastocyst perimeter was larger in the agBL group than in the ivfBL and sBL groups (1168.8 ± 200.23 vs. 887.33 ± 187.62 and 678 ± 226.1 μm; P < 0.05). In addition, mitochondrial fluorescence intensity was higher in the agBL group than in the ivfBL and sBL groups (P < 0.05). The number of apoptotic cells per blastocyst was lower in the ivfBL and agBL groups than in the sBL group (3.7 ± 2.2 and 3.4 ± 2.1 vs. 6.7 ± 6.8; P < 0.05). The genes identified in the microarray belonged to 18 categories. Expression of the Krüppel-like factor 4 gene, which is associated with cell proliferation, development, and transcription, was 7.2-fold higher in the agBL group than in the ivfBL group (P < 0.05) but did not differ between the sBL and ivfBL groups (P > 0.05). Expression of the heat shock 70-kDa protein 1A gene, which is associated with apoptosis, was 12-fold higher in the s

  20. Innovative interactive flexible docking method for multi-scale reconstruction elucidates dystrophin molecular assembly.

    PubMed

    Molza, A-E; Férey, N; Czjzek, M; Le Rumeur, E; Hubert, J-F; Tek, A; Laurent, B; Baaden, M; Delalande, O

    2014-01-01

    At present, our molecular knowledge of dystrophin, the protein encoded by the DMD gene and mutated in myopathy patients, remains limited. To get around the absence of its atomic structure, we have developed an innovative interactive docking method based on the BioSpring software in combination with Small-angle X-ray Scattering (SAXS) data. BioSpring allows interactive handling of biological macromolecules thanks to an augmented Elastic Network Model (aENM) that combines the spring network with non-bonded terms between atoms or pseudo-atoms. This approach can be used for building molecular assemblies even on a desktop or a laptop computer thanks to code optimizations including parallel computing and GPU programming. By combining atomistic and coarse-grained models, the approach significantly simplifies the set-up of multi-scale scenarios. BioSpring is remarkably efficient for the preparation of numeric simulations or for the design of biomolecular models integrating qualitative experimental data restraints. The combination of this program and SAXS allowed us to propose the first high-resolution models of the filamentous central domain of dystrophin, covering repeats 11 to 17. Low-resolution interactive docking experiments driven by a potential grid enabled us to propose how dystrophin may associate with F-actin and nNOS. This information provides an insight into medically relevant discoveries to come. PMID:25340652

  1. Recent innovations in protein separation on microchips by electrophoretic methods: an update.

    PubMed

    Tran, Nguyet Thuy; Ayed, Ichraf; Pallandre, Antoine; Taverna, Myriam

    2010-01-01

    Lab-on-a-chip electrophoresis is becoming increasingly useful for protein analysis, thanks to recent developments in this field. This review is an update of the review we published at the start of 2008 [Peng, Y., Pallandre, A., Tran, N. T., Taverna, M., Electrophoresis 2008, 29, 156-177]. The superiority of polymers for the manufacture of analytical microchips has been confirmed. This trend implies several modifications to the processes previously used with glass/silicon chips and requires a better understanding of the interfacial phenomena of these materials. Significant progress in chip-based techniques for protein analysis has been made in the last 2 years. In addition to advances in traditional electrokinetic modes, counter-flow gradient focusing techniques have emerged as useful methods not only for separation, but also for the online preconcentration of samples. This review, with more than 175 references, presents recent advances and novel strategies for EOF measurement, surface treatment, sample pretreatment, detection and innovations relating to the different modes of separation. PMID:20014053

  2. Innovations and trends in meat consumption: an application of the Delphi method in Spain.

    PubMed

    Chamorro, Antonio; Miranda, Francisco J; Rubio, Sergio; Valero, Víctor

    2012-12-01

    The meat sector in Spain is an important industry. However, traditional consumption is changing as a result of the country's economic crisis and the new structure of households. The objective of the present study was to identify trends in meat consumption in Spain extrapolated to 2016, and the main innovations that should be of interest to firms in the sector. The study was conducted in 2011 using the Delphi method with the participation of 26 experts. The results showed that, while the demand for meat will not vary significantly in amount, it will do so in composition, with chicken replacing beef as the meat of most importance in the shopping basket. In addition, significant growth is expected in certified meat, but the demand for organic meat will not take off. Neither will there be no significant changes in end purchase formats, but there will be a clear trend in consumers' purchasing decision criteria away from price, external appearance and origin towards quality certification and the attributes of the packaging. With respect to end purchase channels, the experts estimate that the current trend will be accentuated with increasing market share for large supermarkets and major distribution brands.

  3. Innovations and trends in meat consumption: an application of the Delphi method in Spain.

    PubMed

    Chamorro, Antonio; Miranda, Francisco J; Rubio, Sergio; Valero, Víctor

    2012-12-01

    The meat sector in Spain is an important industry. However, traditional consumption is changing as a result of the country's economic crisis and the new structure of households. The objective of the present study was to identify trends in meat consumption in Spain extrapolated to 2016, and the main innovations that should be of interest to firms in the sector. The study was conducted in 2011 using the Delphi method with the participation of 26 experts. The results showed that, while the demand for meat will not vary significantly in amount, it will do so in composition, with chicken replacing beef as the meat of most importance in the shopping basket. In addition, significant growth is expected in certified meat, but the demand for organic meat will not take off. Neither will there be no significant changes in end purchase formats, but there will be a clear trend in consumers' purchasing decision criteria away from price, external appearance and origin towards quality certification and the attributes of the packaging. With respect to end purchase channels, the experts estimate that the current trend will be accentuated with increasing market share for large supermarkets and major distribution brands. PMID:22878051

  4. Innovative interactive flexible docking method for multi-scale reconstruction elucidates dystrophin molecular assembly.

    PubMed

    Molza, A-E; Férey, N; Czjzek, M; Le Rumeur, E; Hubert, J-F; Tek, A; Laurent, B; Baaden, M; Delalande, O

    2014-01-01

    At present, our molecular knowledge of dystrophin, the protein encoded by the DMD gene and mutated in myopathy patients, remains limited. To get around the absence of its atomic structure, we have developed an innovative interactive docking method based on the BioSpring software in combination with Small-angle X-ray Scattering (SAXS) data. BioSpring allows interactive handling of biological macromolecules thanks to an augmented Elastic Network Model (aENM) that combines the spring network with non-bonded terms between atoms or pseudo-atoms. This approach can be used for building molecular assemblies even on a desktop or a laptop computer thanks to code optimizations including parallel computing and GPU programming. By combining atomistic and coarse-grained models, the approach significantly simplifies the set-up of multi-scale scenarios. BioSpring is remarkably efficient for the preparation of numeric simulations or for the design of biomolecular models integrating qualitative experimental data restraints. The combination of this program and SAXS allowed us to propose the first high-resolution models of the filamentous central domain of dystrophin, covering repeats 11 to 17. Low-resolution interactive docking experiments driven by a potential grid enabled us to propose how dystrophin may associate with F-actin and nNOS. This information provides an insight into medically relevant discoveries to come.

  5. Innovative Field Methods for Characterizing the Hydraulic Properties of a Complex Fractured Rock Aquifer (Ploemeur, Brittany)

    NASA Astrophysics Data System (ADS)

    Bour, O.; Le Borgne, T.; Longuevergne, L.; Lavenant, N.; Jimenez-Martinez, J.; De Dreuzy, J. R.; Schuite, J.; Boudin, F.; Labasque, T.; Aquilina, L.

    2014-12-01

    Characterizing the hydraulic properties of heterogeneous and complex aquifers often requires field scale investigations at multiple space and time scales to better constrain hydraulic property estimates. Here, we present and discuss results from the site of Ploemeur (Brittany, France) where complementary hydrological and geophysical approaches have been combined to characterize the hydrogeological functioning of this highly fractured crystalline rock aquifer. In particular, we show how cross-borehole flowmeter tests, pumping tests and frequency domain analysis of groundwater levels allow quantifying the hydraulic properties of the aquifer at different scales. In complement, we used groundwater temperature as an excellent tracer for characterizing groundwater flow. At the site scale, measurements of ground surface deformation through long-base tiltmeters provide robust estimates of aquifer storage and allow identifying the active structures where groundwater pressure changes occur, including those acting during recharge process. Finally, a numerical model of the site that combines hydraulic data and groundwater ages confirms the geometry of this complex aquifer and the consistency of the different datasets. The Ploemeur site, which has been used for water supply at a rate of about 106 m3 per year since 1991, belongs to the French network of hydrogeological sites H+ and is currently used for monitoring groundwater changes and testing innovative field methods.

  6. INNOVATIVE METHODS FOR EMISSION-INVENTORY DEVELOPMENT AND EVALUATION: WORKSHOP SUMMARY

    EPA Science Inventory

    Emission inventories are an essential tool for evaluating, managing, and regulating air pollution. Refinements and innovations in instruments that measure air pollutants, models that calculate emissions as well as techniques for data management and uncertainty assessment are nee...

  7. Genes galore: a summary of methods for accessing results from large-scale partial sequencing of anonymous Arabidopsis cDNA clones.

    PubMed Central

    Newman, T; de Bruijn, F J; Green, P; Keegstra, K; Kende, H; McIntosh, L; Ohlrogge, J; Raikhel, N; Somerville, S; Thomashow, M

    1994-01-01

    High-throughput automated partial sequencing of anonymous cDNA clones provides a method to survey the repertoire of expressed genes from an organism. Comparison of the coding capacity of these expressed sequence tags (ESTs) with the sequences in the public data bases results in assignment of putative function to a significant proportion of the ESTs. Thus, the more than 13,400 plant ESTs that are currently available provide a new resource that will facilitate progress in many areas of plant biology. These opportunities are illustrated by a description of the results obtained from analysis of 1500 Arabidopsis ESTs from a cDNA library prepared from equal portions of poly(A+) mRNA from etiolated seedlings, roots, leaves, and flowering inflorescences. More than 900 different sequences were represented, 32% of which showed significant nucleotide or deduced amino acid sequences similarity to previously characterized genes or proteins from a wide range of organisms. At least 165 of the clones had significant deduced amino acid sequence homology to proteins or gene products that have not been previously characterized from higher plants. A summary of methods for accessing the information and materials generated by the Arabidopsis cDNA sequencing project is provided. PMID:7846151

  8. Managing ground and surface water resources using innovative methods in rural and urban areas of Sub-Saharan Africa

    NASA Astrophysics Data System (ADS)

    Mapani, Benjamin; Magole, Lapologang; Makurira, Hodson; Mazvimavi, Dominic; Meck, Maideyi; Mul, Maloes

    2016-06-01

    Management of water resources across the world is becoming more challenging as a result of population increase and the changes in climatic patterns that are now evident across the globe, especially so in Southern Africa. It is therefore imperative that water researchers begin to apply innovative methods that are accurate and reliable. In this editorial we highlight some of the methods that have been applied in this changing environment.

  9. Comparative quantitative analysis of BCR-ABL transcripts with the T315I mutant clone by polymerase chain reaction (PCR)-Invader method.

    PubMed

    Tadokoro, Kenichi; Ishikawa, Maho; Suzuki, Makoto; Saito, Tomoyoshi; Suzuki, Yoshie; Yamaguchi, Toshikazu; Yagasaki, Fumiharu

    2011-09-01

    Drug resistance is a serious complication in the treatment of chronic myeloid leukemia (CML). The most common and best-characterized mechanism of secondary imatinib resistance in CML is the development of kinase domain mutations in the BCR-ABL gene. Second-generation tyrosine kinase inhibitors, such as dasatinib or nilotinib, overcome most of these mutations, but they are not effective against the T315I mutant. To determine whether these mutations contribute to clinical resistance, it is necessary to monitor the ratio of the mutant and wild-type forms. Here, we developed a polymerase chain reaction (PCR)-Invader assay for comparative quantitative analysis (qPI assay) of BCR-ABL transcripts with the T315I mutant clone. T315I ratios were calculated for the wild-type and mutant fold-over-zero (FOZ) values. In examination with 2 kinds of plasmids containing wild-type or T315I mutant PCR amplicons, mutant FOZ values were detected down to 1% of the total. The results of 12 serial samples from 2 patients (case A: Philadelphia-positive acute lymphoblastic leukemia and case B: CML) with the T315I mutant clone were compared with those of direct sequencing or 2 kinds of allele-specific oligonucleotide (ASO)-PCR. All samples showed the T315I mutation by qPI assay and ASO-PCR, and 10 samples showed it by direct sequencing. Significant correlation (correlation coefficient; r2 = 0.951) was noted between the qPI assay and quantitative ASO-PCR to analyze T315I mutant ratios. Thus, the qPI assay is a useful method for evaluating the T315I mutant clone in BCR-ABL transcripts.

  10. Comparative quantitative analysis of BCR-ABL transcripts with the T315I mutant clone by polymerase chain reaction (PCR)-Invader method.

    PubMed

    Tadokoro, Kenichi; Ishikawa, Maho; Suzuki, Makoto; Saito, Tomoyoshi; Suzuki, Yoshie; Yamaguchi, Toshikazu; Yagasaki, Fumiharu

    2011-09-01

    Drug resistance is a serious complication in the treatment of chronic myeloid leukemia (CML). The most common and best-characterized mechanism of secondary imatinib resistance in CML is the development of kinase domain mutations in the BCR-ABL gene. Second-generation tyrosine kinase inhibitors, such as dasatinib or nilotinib, overcome most of these mutations, but they are not effective against the T315I mutant. To determine whether these mutations contribute to clinical resistance, it is necessary to monitor the ratio of the mutant and wild-type forms. Here, we developed a polymerase chain reaction (PCR)-Invader assay for comparative quantitative analysis (qPI assay) of BCR-ABL transcripts with the T315I mutant clone. T315I ratios were calculated for the wild-type and mutant fold-over-zero (FOZ) values. In examination with 2 kinds of plasmids containing wild-type or T315I mutant PCR amplicons, mutant FOZ values were detected down to 1% of the total. The results of 12 serial samples from 2 patients (case A: Philadelphia-positive acute lymphoblastic leukemia and case B: CML) with the T315I mutant clone were compared with those of direct sequencing or 2 kinds of allele-specific oligonucleotide (ASO)-PCR. All samples showed the T315I mutation by qPI assay and ASO-PCR, and 10 samples showed it by direct sequencing. Significant correlation (correlation coefficient; r2 = 0.951) was noted between the qPI assay and quantitative ASO-PCR to analyze T315I mutant ratios. Thus, the qPI assay is a useful method for evaluating the T315I mutant clone in BCR-ABL transcripts. PMID:21867983

  11. Steel Casing Resistivity Technology (SCRT): Innovative Applications of Electrical Methods for Buried Tank Leak Detection

    NASA Astrophysics Data System (ADS)

    Fink, J. B.; Levitt, M. T.; Gee, G. W.

    2002-12-01

    The need for non-invasive leak detection methods is extremely important for monitoring cleanup efforts of nuclear waste contained in underground storage tanks at the Hanford Nuclear Facility in Washington. Drilling is both very expensive and undesirable in the tank farms. Various geophysical imaging methods were evaluated over the past two years at two "cold" sites but within geologic conditions similar to the tank farms. The "cold" sites consisted of 1. a dense array of 32 steel casings, and 2. a "mock tank" in which various controlled leaks (injections) of a saturated aqueous solution of Na2S2O35H20 were metered. Nearly all methods required invasive drilling for subsurface placement of sensors. An innovative direct-current electrical method using existing infrastructure as grounding electrodes, such as steel casings and steel tanks, has shown very promising results and is undergoing further testing. The most useful results have been obtained by using multiple grounding points for spatial determinations and continuous time-series monitoring for temporal variations. Although the large size of tanks and lengths of casings make discrete volume estimations difficult, data acquired for test leaks to date have shown a surprising correlation between leak rates and the rate-of-change of specific electrical measurements. First order volume approximations can be made based on existing knowledge of the geologic environment and hydraulic parameters. Spatial data provide general leak location and gross flow characteristics, whereas temporal data indicate test leak commencement, cessation, and approximate leak rates. On-going testing is providing quantitative calibration information that is expected to transfer to the tank farm environments. Procedures are being developed that will simplify the installation and operation of the system in the tank-farm environments. Implementation of the technology necessitated remote operation and monitoring of the electrical system

  12. On classical cloning and no-cloning

    NASA Astrophysics Data System (ADS)

    Teh, Nicholas J.

    2012-02-01

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

  13. Overlap extension PCR cloning.

    PubMed

    Bryksin, Anton; Matsumura, Ichiro

    2013-01-01

    Rising demand for recombinant proteins has motivated the development of efficient and reliable cloning methods. Here we show how a beginner can clone virtually any DNA insert into a plasmid of choice without the use of restriction endonucleases or T4 DNA ligase. Chimeric primers encoding plasmid sequence at the 5' ends and insert sequence at the 3' ends are designed and synthesized. Phusion(®) DNA polymerase is utilized to amplify the desired insert by PCR. The double-stranded product is subsequently employed as a pair of mega-primers in a PCR-like reaction with circular plasmids. The original plasmids are then destroyed in restriction digests with Dpn I. The product of the overlap extension PCR is used to transform competent Escherichia coli cells. Phusion(®) DNA polymerase is used for both the amplification and fusion reactions, so both steps can be monitored and optimized in the same way. PMID:23996437

  14. Fluorescence in situ hybridization of 16S rRNA gene clones (Clone-FISH) for probe validation and screening of clone libraries.

    PubMed

    Schramm, Andreas; Fuchs, Bernhard M; Nielsen, Jeppe L; Tonolla, Mauro; Stahl, David A

    2002-11-01

    A method is presented for fluorescence in situ hybridization (FISH) of 16S rRNA gene clones targeting in vivo transcribed plasmid inserts (Clone-FISH). Several different cloning approaches and treatments to generate target-rRNA in the clones were compared. Highest signal intensities of Clone-FISH were obtained using plasmids with a T7 RNA polymerase promoter and host cells with an IPTG-inducible T7 RNA polymerase. Combined IPTG-induction and chloramphenicol treatment of those clones resulted in FISH signals up to 2.8-fold higher than signals of FISH with probe EUB338 to cells of Escherichia coli. Probe dissociation curves for three oligonucleotide probes were compared for reference cells containing native (FISH) or cloned (Clone-FISH) target sequences. Melting behaviour and calculated T(d) values were virtually identical for clones and cells, providing a format to use 16S rRNA gene clones instead of pure cultures for probe validation and optimization of hybridization conditions. The optimized Clone-FISH protocol was also used to screen an environmental clone library for insert sequences of interest. In this application format, 13 out of 82 clones examined were identified to contain sulphate-reducing bacterial rRNA genes. In summary, Clone-FISH is a simple and fast technique, compatible with a wide variety of cloning vectors and hosts, that should have general utility for probe validation and screening of clone libraries. PMID:12460279

  15. Innovative method and apparatus for the deep cleaning of soluble salts from mortars and lithic materials

    NASA Astrophysics Data System (ADS)

    Gaggero, Laura; Ferretti, Maurizio; Torrielli, Giulia; Caratto, Valentina

    2016-04-01

    demonstrated by conducibility tests on the juxtaposed Japanese paper. In addition, after the conventional treatment, a considerable amount of soluble salts was further extracted demonstrating that traditional wraps operate just a shallow cleaning, and soluble salts are liable to emerge later as efflorescence affecting the conservation after restoration. The optimum cleaning was obtained by finishing the innovative extraction with sepiolite/cellulose wraps. As a whole, the novel method and apparatus enhance the time for restoration and the final quality before consolidation and protection. [1] "Apparatus and method for treating porous materials" - M. Ferretti, L. Gaggero, G. Torrielli, PCT/IB2015/055129 (2015)

  16. a Simplified Parameter Design Method for Transformation Optics-Based Metamaterial Innovative Cloak

    NASA Astrophysics Data System (ADS)

    Li, Ting-Hua; Huang, Ming; Yang, Jing-Jing; Lu, Jin; Cao, Hui-Lu

    2013-10-01

    Transformation optics-based innovative cloak which combines the virtues of both internal and external cloaks to enable arbitrary multi-objects hidden with visions and movements was first proposed by Huang et al. [Appl. Phys. Lett.101, 151901 (2012)]. But it is rather difficult to implement in practice, for the required material parameters vary with radius and even have singular values. To accelerate its practical realization but still keep good performance of invisibility, a simplified innovative cloak with only spatially varying axial parameter is developed via choosing appropriate transformation function. The advantage of such a cloak is that both radial and azimuthal parameters are constants, and all three components are nonsingular and finite. Full-wave simulation confirms the perfect cloaking effect of the cloak. Besides, the influences of metamaterials loss and parameter deviation on the performance of cloak are also investigated. This work provides a simple and feasible solution to push metamaterial-assisted innovative cloak more closely to the practice.

  17. The Clone Factory

    ERIC Educational Resources Information Center

    Stoddard, Beryl

    2005-01-01

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

  18. Human cloning 2001.

    PubMed

    Healy, David L; Weston, Gareth; Pera, Martin F; Rombauts, Luk; Trounson, Alan O

    2002-05-01

    This review summaries human cloning from a clinical perspective. Natural human clones, that is, monozygotic twins, are increasing in the general community. Iatrogenic human clones have been produced for decades in infertile couples given fertility treatment such as ovulation induction. A clear distinction must be made between therapeutic cloning using embryonic stem cells and reproductive cloning attempts. Unlike the early clinical years of in vitro fertilization, with cloning there is no animal model that is safe and dependable. Until there is such a model, 'Dolly'-style human cloning is medically unacceptable.

  19. An Innovative Method for Preparing Semiconductor Change Used in Crystal Growth and Shear Cell Diffusion Experiments

    NASA Technical Reports Server (NTRS)

    Anrold, William A.; Matthiesen, David; Benett, Robert J.; Jayne, Douglas T.

    1997-01-01

    An innovative technique for machining semiconductors has been developed. This technique was used to prepare semiconductor charges for crystal growth and shear cell diffusion experiments. The technique allows brittle semiconductor materials to be quickly and accurately machined. Lightly doping the semiconductor material increases the conductivity enough to allow the material to be shaped by an electrical discharge machine (EDM).

  20. Integrating Teacher- and Peer-Assessments of Group Coursework Assignments in Business Education: Some Innovative Methods

    ERIC Educational Resources Information Center

    Onyia, Okey Peter

    2014-01-01

    This paper is a sequel to an earlier one that examines "the efficacy of two innovative peer-assessment templates ("PET" and "PACT") introduced to enable students provide evidence of their fairness in evaluating peer contributions to group project work" (Onyia, O. P. and Allen, S., 2012). In the present paper, three…

  1. Building America Top Innovations 2012: Building Energy Optimization Analysis Method (BEopt)

    SciTech Connect

    none,

    2013-01-01

    This Building America Top Innovations profile describes the DOE-sponsored BEopt software, which ensures a consistent analysis platform and accurate simulations. Many BEopt algorithms have been adopted by private-sector HERS software tools that have helped improve the energy efficiency of tens-of-thousands of ENERGY STAR-certified homes.

  2. Innovation and a Return to the Status: A Mixed-Methods Study of School Reconstitution

    ERIC Educational Resources Information Center

    Strunk, Katharine O.; Marsh, Julie A.; Hashim, Ayesha K.; Bush-Mecenas, Susan

    2016-01-01

    School reconstitution, a turnaround strategy that prescribes massive staffing turnover, is expected to result in more committed and capable school staff and innovative practices. However, little evidence supports this assumption. We use quasi-experimental designs to assess the impact of reconstitution on student achievement and teacher mobility,…

  3. Innovation in Postgraduate Teaching: Mixed Methods to Enhance Learning and Learning about Learning

    ERIC Educational Resources Information Center

    Dickie, Carolyn; Jay, Leighton

    2010-01-01

    Growing pressure to restructure and reform tertiary education is encouraging university academics to use innovative practices that assist students to develop "employable" skills. The hybrid approach described in this paper stimulated students to be self-directed adult learners who maximized their learning of content and skills by means of…

  4. Innovative methods to stabilize liquid membranes for removal of radionuclides from groundwater

    SciTech Connect

    Lokhandwala, K.

    1997-10-01

    In this Phase I Small Business Innovation Research program, Membrane Technology Research, Inc., is developing a stable liquid membrane for extracting uranium and other radionuclides from groundwater. The improved membrane can also be applied to separation of other metal ions from aqueous streams in industrial operations.

  5. Sequential cloning of chromosomes

    DOEpatents

    Lacks, Sanford A.

    1995-07-18

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

  6. Sequential cloning of chromosomes

    DOEpatents

    Lacks, S.A.

    1995-07-18

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

  7. Methods for open innovation on a genome-design platform associating scientific, commercial, and educational communities in synthetic biology.

    PubMed

    Toyoda, Tetsuro

    2011-01-01

    Synthetic biology requires both engineering efficiency and compliance with safety guidelines and ethics. Focusing on the rational construction of biological systems based on engineering principles, synthetic biology depends on a genome-design platform to explore the combinations of multiple biological components or BIO bricks for quickly producing innovative devices. This chapter explains the differences among various platform models and details a methodology for promoting open innovation within the scope of the statutory exemption of patent laws. The detailed platform adopts a centralized evaluation model (CEM), computer-aided design (CAD) bricks, and a freemium model. It is also important for the platform to support the legal aspects of copyrights as well as patent and safety guidelines because intellectual work including DNA sequences designed rationally by human intelligence is basically copyrightable. An informational platform with high traceability, transparency, auditability, and security is required for copyright proof, safety compliance, and incentive management for open innovation in synthetic biology. GenoCon, which we have organized and explained here, is a competition-styled, open-innovation method involving worldwide participants from scientific, commercial, and educational communities that aims to improve the designs of genomic sequences that confer a desired function on an organism. Using only a Web browser, a participating contributor proposes a design expressed with CAD bricks that generate a relevant DNA sequence, which is then experimentally and intensively evaluated by the GenoCon organizers. The CAD bricks that comprise programs and databases as a Semantic Web are developed, executed, shared, reused, and well stocked on the secure Semantic Web platform called the Scientists' Networking System or SciNetS/SciNeS, based on which a CEM research center for synthetic biology and open innovation should be established.

  8. Cloned plasmid DNA fragments as calibrators for controlling GMOs: different real-time duplex quantitative PCR methods.

    PubMed

    Taverniers, Isabel; Van Bockstaele, Erik; De Loose, Marc

    2004-03-01

    Analytical real-time PCR technology is a powerful tool for implementation of the GMO labeling regulations enforced in the EU. The quality of analytical measurement data obtained by quantitative real-time PCR depends on the correct use of calibrator and reference materials (RMs). For GMO methods of analysis, the choice of appropriate RMs is currently under debate. So far, genomic DNA solutions from certified reference materials (CRMs) are most often used as calibrators for GMO quantification by means of real-time PCR. However, due to some intrinsic features of these CRMs, errors may be expected in the estimations of DNA sequence quantities. In this paper, two new real-time PCR methods are presented for Roundup Ready soybean, in which two types of plasmid DNA fragments are used as calibrators. Single-target plasmids (STPs) diluted in a background of genomic DNA were used in the first method. Multiple-target plasmids (MTPs) containing both sequences in one molecule were used as calibrators for the second method. Both methods simultaneously detect a promoter 35S sequence as GMO-specific target and a lectin gene sequence as endogenous reference target in a duplex PCR. For the estimation of relative GMO percentages both "delta C(T)" and "standard curve" approaches are tested. Delta C(T) methods are based on direct comparison of measured C(T) values of both the GMO-specific target and the endogenous target. Standard curve methods measure absolute amounts of target copies or haploid genome equivalents. A duplex delta C(T) method with STP calibrators performed at least as well as a similar method with genomic DNA calibrators from commercial CRMs. Besides this, high quality results were obtained with a standard curve method using MTP calibrators. This paper demonstrates that plasmid DNA molecules containing either one or multiple target sequences form perfect alternative calibrators for GMO quantification and are especially suitable for duplex PCR reactions.

  9. Multipartite asymmetric quantum cloning

    SciTech Connect

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

    2005-10-15

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

  10. Ethical issues in cloning.

    PubMed

    Satris, S

    2000-01-01

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

  11. The identity of clones.

    PubMed

    Evers, K

    1999-02-01

    A common concern with respect to cloning is based on the belief that cloning produces identical individuals. This is a fundamental misunderstanding of what type of identity-relation cloning involves. The concept "identity" is ambiguous, and the statement that cloning produces "identical" individuals is not meaningful unless the notion of identity is clarified. This paper distinguishes between numerical and qualitative; relational and intrinsic: logical and empirical identity, and discusses the empirical individuation of clones in terms of genetics, physiology, perception, cognition and personality. I argue that the only relation of identity cloning involves is qualitative, intrinsic and empirical: genetic indiscernibility, unlikely to include identity under other aspects mentioned. A popular argument against cloning claims our "right" to a "unique identity". This objection either implies (absurdly) the right not to be an identical twin, or assumes (incorrectly) that cloning involves identity other than genetic. Either way, the argument is untenable.

  12. Aristotle and headless clones.

    PubMed

    Mosteller, Timothy

    2005-01-01

    Cloned organisms can be genetically altered so that they do not exhibit higher brain functioning. This form of therapeutic cloning allows for genetically identical organs and tissues to be harvested from the clone for the use of the organism that is cloned. "Spare parts" cloning promises many opportunities for future medical advances. What is the ontological and ethical status of spare parts, headless clones? This paper attempts to answer this question from the perspective of Aristotle's view of the soul. Aristotle's metaphysics as applied to his view of biological essences generates an ethic that can contribute to moral reasoning regarding the use of headless spare parts clones. The task of this paper is to show the implications that Aristotle's view of the soul, if it is true, would have on the ethics of headless, spare parts cloning. PMID:16180113

  13. Aristotle and headless clones.

    PubMed

    Mosteller, Timothy

    2005-01-01

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

  14. Best practices: implementation of a glucose screening program based on diffusion of innovation theory methods.

    PubMed

    Nicol, Ginger E; Morrato, Elaine H; Johnson, Mark C; Campagna, Elizabeth; Yingling, Michael D; Pham, Victor; Newcomer, John W

    2011-01-01

    There is public health interest in the identification and treatment of modifiable cardiometabolic risk factors among patients treated with antipsychotic medications. However, best-practice screening recommendations endorsed by multiple medical organizations have not translated into real-world clinical practice. Quality improvement strategies may help to address the gap between policy and implementation. This column describes the successful implementation of a best-practice glucose screening program in a large network of community mental health centers that was based on Six Sigma and diffusion of innovation theory.

  15. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    PubMed Central

    Beyer, Hannes M.; Gonschorek, Patrick; Samodelov, Sophia L.; Meier, Matthias; Weber, Wilfried; Zurbriggen, Matias D.

    2015-01-01

    Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly), a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date. PMID:26360249

  16. Incorporating café design principles into End-of-Life discussions: an innovative method for continuing education.

    PubMed

    Kanaskie, Mary Louise

    2011-04-01

    Café design provides an innovative method for conducting continuing education activities. This method was chosen to elicit meaningful conversation based on issues related to End-of-Life care. Café design principles incorporate the following: setting the context, creating hospitable space, exploring questions that matter, encouraging everyone's contributions, connecting diverse perspectives, listening together for insights, and sharing collective discoveries. Key discussion questions were identified from the End-of Life Nursing Education Consortium Core Curriculum. Questions were revised to incorporate the principles of appreciative inquiry, which encourage a shift from traditional methods of problem identification to creation of a positive vision. Participants rated the café design method as an effective way to share their ideas and to stimulate conversation.

  17. Incorporating café design principles into End-of-Life discussions: an innovative method for continuing education.

    PubMed

    Kanaskie, Mary Louise

    2011-04-01

    Café design provides an innovative method for conducting continuing education activities. This method was chosen to elicit meaningful conversation based on issues related to End-of-Life care. Café design principles incorporate the following: setting the context, creating hospitable space, exploring questions that matter, encouraging everyone's contributions, connecting diverse perspectives, listening together for insights, and sharing collective discoveries. Key discussion questions were identified from the End-of Life Nursing Education Consortium Core Curriculum. Questions were revised to incorporate the principles of appreciative inquiry, which encourage a shift from traditional methods of problem identification to creation of a positive vision. Participants rated the café design method as an effective way to share their ideas and to stimulate conversation. PMID:21656936

  18. Cloning. Pigs is pigs.

    PubMed

    Prather, R S

    2000-09-15

    Since the first report of a cloned animal (Dolly the sheep) 3 years ago, cloning mammals has become something of a cottage industry. As Prather discusses in his Perspective, pigs can now be added to the august list of cloned animals, which includes cows, goats, and mice. This is a particularly spectacular achievement because pig cloning has turned out to be notoriously difficult. The pig is also a valuable domestic animal to have cloned because, being physiologically close to humans, its organs can be used in xenotransplantation.

  19. Cloning. Pigs is pigs.

    PubMed

    Prather, R S

    2000-09-15

    Since the first report of a cloned animal (Dolly the sheep) 3 years ago, cloning mammals has become something of a cottage industry. As Prather discusses in his Perspective, pigs can now be added to the august list of cloned animals, which includes cows, goats, and mice. This is a particularly spectacular achievement because pig cloning has turned out to be notoriously difficult. The pig is also a valuable domestic animal to have cloned because, being physiologically close to humans, its organs can be used in xenotransplantation. PMID:11012362

  20. [Mystery and problems of cloning].

    PubMed

    Nikitin, V A

    2010-01-01

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

  1. Innovative test method for the estimation of the foaming tendency of substrates for biogas plants.

    PubMed

    Moeller, Lucie; Eismann, Frank; Wißmann, Daniel; Nägele, Hans-Joachim; Zielonka, Simon; Müller, Roland A; Zehnsdorf, Andreas

    2015-07-01

    Excessive foaming in anaerobic digestion occurs at many biogas plants and can cause problems including plugged gas pipes. Unfortunately, the majority of biogas plant operators are unable to identify the causes of foaming in their biogas reactor. The occurrence of foaming is often related to the chemical composition of substrates fed to the reactor. The consistency of the digestate itself is also a crucial part of the foam formation process. Thus, no specific recommendations concerning substrates can be given in order to prevent foam formation in biogas plants. The safest way to avoid foaming is to test the foaming tendency of substrates on-site. A possible solution is offered by an innovative foaming test. With the help of this tool, biogas plant operators can evaluate the foaming disposition of new substrates prior to use in order to adjust the composition of substrate mixes.

  2. An Innovative Hybrid Loop-Pool SFR Design and Safety Analysis Methods: Today and Tomorrow

    SciTech Connect

    Hongbin Zhang; Haihua Zhao; Vincent Mousseau

    2008-04-01

    Investment in commercial sodium cooled fast reactor (SFR) power plants will become possible only if SFRs achieve economic competitiveness as compared to light water reactors and other Generation IV reactors. Toward that end, we have launched efforts to improve the economics and safety of SFRs from the thermal design and safety analyses perspectives at Idaho National Laboratory. From the thermal design perspective, an innovative hybrid loop-pool SFR design has been proposed. This design takes advantage of the inherent safety of a pool design and the compactness of a loop design to further improve economics and safety. From the safety analyses perspective, we have initiated an effort to develop a high fidelity reactor system safety code.

  3. The Facial Contouring and Support System: An Innovative Method for Midfacial Fat Repositioning

    PubMed Central

    2014-01-01

    Summary: Minimally invasive rejuvenation procedures are increasingly popular with patients. In the midface, these might involve the introduction of sutures to lift and secure the malar tissue, fat grafts, and fillers to increase volume. This article describes a new facial contouring and support system, which uses an innovative hollow, double-beveled needle to which a 2/0 polypropylene suture may be anchored. Among 102 patients there were no complications, and follow-up at 3–5 years indicates little or no loss of satisfaction with the outcome. The procedure can be combined with other modalities—including fillers and skin peels—to achieve an overall, balanced, natural look for the patient. Potential adjustability and reversibility of the procedure are reassuring for both patient and surgeon and add to the technique’s versatility. PMID:25426398

  4. Self-Cloning CRISPR.

    PubMed

    Arbab, Mandana; Sherwood, Richard 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 recombines with short PCR-amplified site-specific sgRNA sequences within the target cell by homologous recombination to circumvent the process of sgRNA plasmid construction. Through this mechanism, scCRISPR enables gene editing within 2 hr once sgRNA oligos are available, with high efficiency equivalent to conventional sgRNA targeting: >90% gene knockout in both mouse and human embryonic stem cells and cancer cell lines. Furthermore, using PCR-based addition of short homology arms, we achieve efficient site-specific knock-in of transgenes such as GFP without traditional plasmid cloning or genome-integrated selection cassette (2% to 4% knock-in rate). The methods in this paper describe the most rapid and efficient means of CRISPR gene editing. © 2016 by John Wiley & Sons, Inc. PMID:27532819

  5. Self-Cloning CRISPR.

    PubMed

    Arbab, Mandana; Sherwood, Richard 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 recombines with short PCR-amplified site-specific sgRNA sequences within the target cell by homologous recombination to circumvent the process of sgRNA plasmid construction. Through this mechanism, scCRISPR enables gene editing within 2 hr once sgRNA oligos are available, with high efficiency equivalent to conventional sgRNA targeting: >90% gene knockout in both mouse and human embryonic stem cells and cancer cell lines. Furthermore, using PCR-based addition of short homology arms, we achieve efficient site-specific knock-in of transgenes such as GFP without traditional plasmid cloning or genome-integrated selection cassette (2% to 4% knock-in rate). The methods in this paper describe the most rapid and efficient means of CRISPR gene editing. © 2016 by John Wiley & Sons, Inc.

  6. Cloning, expression, purification and characterization of his-tagged human glucose-6-phosphate dehydrogenase: a simplified method for protein yield.

    PubMed

    Gómez-Manzo, Saúl; Terrón-Hernández, Jessica; de la Mora-de la Mora, Ignacio; García-Torres, Itzhel; López-Velázquez, Gabriel; Reyes-Vivas, Horacio; Oria-Hernández, Jesús

    2013-10-01

    Glucose-6-phosphate dehydrogenase (G6PD) catalyzes the first step of the pentose phosphate pathway. In erythrocytes, the functionality of the pathway is crucial to protect these cells against oxidative damage. G6PD deficiency is the most frequent enzymopathy in humans with a global prevalence of 4.9 %. The clinical picture is characterized by chronic or acute hemolysis in response to oxidative stress, which is related to the low cellular activity of G6PD in red blood cells. The disease is heterogeneous at genetic level with around 160 mutations described, mostly point mutations causing single amino acid substitutions. The biochemical studies aimed to describe the detrimental effects of mutations on the functional and structural properties of human G6PD are indispensable to understand the molecular physiopathology of this disease. Therefore, reliable systems for efficient expression and purification of the protein are highly desirable. In this work, human G6PD was heterologously expressed in Escherichia coli and purified by immobilized metal affinity chromatography in a single chromatographic step. The structural and functional characterization indicates that His-tagged G6PD resembles previous preparations of recombinant G6PD. In contrast with previous protein yield systems, our method is based on commonly available resources and fully accessible laboratory equipment; therefore, it can be readily implemented.

  7. Archaeal Diversity of Upland Rice Field Soils Assessed by the Terminal Restriction Fragment Length Polymorphism Method Combined with Real Time Quantitative-PCR and a Clone Library Analysis.

    PubMed

    Nishizawa, Tomoyasu; Komatsuzaki, Masakazu; Kaneko, Nobuhiro; Ohta, Hiroyuki

    2008-01-01

    The PCR amplification-based analysis of microbial diversity is subject to potential problems. In this study, to minimize the bias toward a 1:1 ratio in multitemplate PCR, a real-time PCR assay was carried out using a quenching fluorescence dye primer and amplification efficiency was monitored. Then terminal-restriction fragment length polymorphism (T-RFLP) profiling was performed using the PCR product with minimized PCR bias. This method was applied to an analysis of the diversity of the archaeal community in an upland rice field under different tillage systems and winter cover cropping. Terminal restriction fragments (T-RFs) of PCR-amplified archaeal 16S rRNA genes were assigned to the gene sequences recovered from the same soil by using an archaeal 16S rRNA gene clone library. Our results indicated that soil archaeal members were not influenced but the relative abundance of archaeal species particularly those belonging to Crenarchaeota which changed between the tillage and non-tillage treatments.

  8. The Trial of Drug Discovery using the In-Silico Screening Methods Developed by Pharmaceutical Innovation Value Chain

    NASA Astrophysics Data System (ADS)

    Inoue, Tsuyoshi; Matsumura, Hiroyoshi; Adachi, Hiroaki; Mori, Yusuke; Takano, Kazufumi; Murakami, Satoshi; Fukunishi, Yoshifumi; Nakamura, Haruki; Kinoshita, Takayoshi; Nakanishi, Isao; Okuno, Yasushi; Minakata, Satoshi; Mikami, Yoshiaki; Sakuma, Toshihiro; Kitajima, Masato; Fukuoka, Yoshitada; Takada, Toshikazu; Sakata, Tsuneaki

    We have recently established Pharmaceutical Innovation Value Chain collaborated by The SOSHO project (http://www.so-sho.jp) and The BioGrid Project (http://www.biogrid.jp/) to accelerate new drug development. The in-silico group calculated the matrices on the interaction between the proteins and chemical compounds, and developed the novel in-silico screening methods, Multiple Target Screening (MTS) and Docking score index (DSI), improving the hit rate of screening a lead compound. We have applied these methods for the two target enzymes; human hematopoietic prostaglandin D synthase (H-PGDS) and orotidine 5’-monophosphate decarboxylase from human malaria parasite plasmodium falciparum (PfOMPDC). The optimizing of HQL-79, one of the known inhibitors for human H-PGDS and the screening of lead compounds for both enzymes are in study.

  9. The orbits--anatomical features in view of innovative surgical methods.

    PubMed

    Cornelius, Carl-Peter; Mayer, Peter; Ehrenfeld, Michael; Metzger, Marc Christian

    2014-10-01

    The aim of this article is to update on anatomical key elements of the orbits in reference to surgical innovations. This is a selective literature review supplemented with the personal experience of the authors, using illustrations and photographs of anatomical dissections. The seven osseous components of the orbit can be conceptualized into a simple geometrical layout of a four-sided pyramid with the anterior aditus as a base and the posterior cone as apex. All neurovascular structures pass through bony openings in the sphenoid bone before diversification in the mid and anterior orbit. A set of landmarks such as the optic and maxillary strut comes into new focus. Within the topographical surfaces of the internal orbit the lazy S-shaped floor and the posteromedial bulge are principal determinants for the ocular globe position. The inferomedial orbital strut represents a discernible sagittal buttress. The periorbita and orbital soft tissue contents--extraocular muscles, septae, neurovasculature--are detailed and put into context with periorbital dissection.

  10. Innovative test method for the estimation of the foaming tendency of substrates for biogas plants

    SciTech Connect

    Moeller, Lucie; Eismann, Frank; Wißmann, Daniel; Nägele, Hans-Joachim; Zielonka, Simon; Müller, Roland A.; Zehnsdorf, Andreas

    2015-07-15

    Graphical abstract: Display Omitted - Highlights: • Foaming in biogas plants depends on the interactions between substrate and digestate. • Foaming tests enable the evaluation of substrate foaming tendency in biogas plants. • Leipzig foam tester enables foaming tests of substrates prior to use. - Abstract: Excessive foaming in anaerobic digestion occurs at many biogas plants and can cause problems including plugged gas pipes. Unfortunately, the majority of biogas plant operators are unable to identify the causes of foaming in their biogas reactor. The occurrence of foaming is often related to the chemical composition of substrates fed to the reactor. The consistency of the digestate itself is also a crucial part of the foam formation process. Thus, no specific recommendations concerning substrates can be given in order to prevent foam formation in biogas plants. The safest way to avoid foaming is to test the foaming tendency of substrates on-site. A possible solution is offered by an innovative foaming test. With the help of this tool, biogas plant operators can evaluate the foaming disposition of new substrates prior to use in order to adjust the composition of substrate mixes.

  11. The orbits--anatomical features in view of innovative surgical methods.

    PubMed

    Cornelius, Carl-Peter; Mayer, Peter; Ehrenfeld, Michael; Metzger, Marc Christian

    2014-10-01

    The aim of this article is to update on anatomical key elements of the orbits in reference to surgical innovations. This is a selective literature review supplemented with the personal experience of the authors, using illustrations and photographs of anatomical dissections. The seven osseous components of the orbit can be conceptualized into a simple geometrical layout of a four-sided pyramid with the anterior aditus as a base and the posterior cone as apex. All neurovascular structures pass through bony openings in the sphenoid bone before diversification in the mid and anterior orbit. A set of landmarks such as the optic and maxillary strut comes into new focus. Within the topographical surfaces of the internal orbit the lazy S-shaped floor and the posteromedial bulge are principal determinants for the ocular globe position. The inferomedial orbital strut represents a discernible sagittal buttress. The periorbita and orbital soft tissue contents--extraocular muscles, septae, neurovasculature--are detailed and put into context with periorbital dissection. PMID:25397705

  12. Teaching innovation.

    PubMed

    Lachman, Vicki D; Glasgow, Mary Ellen Smith; Donnelly, Gloria F

    2009-01-01

    Innovation in healthcare is essential to solve the "wicked problems" currently facing healthcare. This article focuses on nature of innovation and how it operates, how innovators think and view problems, how the theory and practice of innovation can be taught in novel ways, and how organizational cultures foster or suppress innovation. Examples of teaching strategies and nurse-driven innovation illustrate the theory and practice of innovation.

  13. Comparative assessment of next-generation sequencing, denaturing gradient gel electrophoresis, clonal restriction fragment length polymorphism and cloning-sequencing as methods for characterizing commercial microbial consortia.

    PubMed

    Samarajeewa, A D; Hammad, A; Masson, L; Khan, I U H; Scroggins, R; Beaudette, L A

    2015-01-01

    Characterization of commercial microbial consortia products for human and environmental health risk assessment is a major challenge for regulatory agencies. As a means to develop an approach to assess the potential environmental risk of these products, research was conducted to compare four genomics methods for characterizing bacterial communities; (i) Denaturing Gradient Gel Electrophoresis (DGGE), (ii) Clonal-Restriction Fragment Length Polymorphism (C/RFLP), (iii) partial 16S rDNA amplification, cloning followed by Sanger sequencing (PRACS) and (iv) Next-Generation Sequencing (NGS) based on Ion Torrent technology. A commercially available microbial consortium, marketed as a remediation agent for degrading petroleum hydrocarbon contamination in soil and water, was assessed. The bacterial composition of the commercial microbial product was characterized using the above four methods. PCR amplification of 16S rDNA was performed targeting the variable region V6 for DGGE, C/RFLP and PRACS and V5 for Ion Torrent sequencing. Ion Torrent technology was shown to be a promising tool for initial screening by detecting the majority of bacteria in the consortium that were also detected by DGGE, C/RFLP and PRACS. Additionally, Ion Torrent sequencing detected some of the bacteria that were claimed to be in the product, while three other methods failed to detect these specific bacteria. However, the relative proportions of the microbial composition detected by Ion Torrent were found to be different from DGGE, C/RFLP and PRACS, which gave comparable results across these three methods. The discrepancy of the Ion Torrent results may be due to the short read length generated by this technique and the targeting of different variable regions on the 16S rRNA gene used in this study. Arcobacter spp. a potential pathogenic bacteria was detected in the product by all methods, which was further confirmed using genus and species-specific PCR, RFLP and DNA-based sequence analyses. However

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

    PubMed

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

    2011-05-01

    We have developed a Single-Tube Restriction-based Ultrafiltration (STRU) cloning procedure that updates traditional ligation-dependent cloning to challenge the newer, faster and more efficient ligation-free techniques and could make it the method of choice. STRU-cloning employs centrifugal filter units with membrane of suitable cut off to remove small unwanted DNA fragments created during restriction of plasmids or PCR products. Heat inactivation, of restriction enzymes, followed by DNA ligation is then performed on the filtrate. By removing the agarose gel electrophoresis DNA purification step from the traditional protocol, which is time consuming and is known to be the cause of ligation problems, STRU-cloning becomes fast, very efficient, inexpensive and offers the highest degree of cloning flexibility by using restriction sites and can be performed in a single tube. This novel agarose gel-free cloning procedure provides benefits for both small and large scale cloning projects. Unlike traditional cloning it can be easily implemented as a fully automated process at very low costs. PMID:21052867

  15. Concept design and hydrodynamic optimization of an innovative SWATH USV by CFD methods

    NASA Astrophysics Data System (ADS)

    Brizzolara, Stefano; Curtin, Tom; Bovio, Marco; Vernengo, Giuliano

    2012-02-01

    The paper presents the main characteristics of an innovative platform which has been conceived and designed to extend the operational capabilities of current unmanned surface vehicles in terms of platform stability in waves and of powering requirement at a relatively high speed. The main idea which rules the project is the realization of a small autonomous surface unit (about 6 m in length) capable of undertaking several tasks in the marine environment even with moderate rough sea conditions. The designed vessel has the ability to locate, recover, and launch other members of the autonomous fleet (like AUVs or other underwater devices) and at the same time could carry out a surveillance service of the surrounding areas. To manage these tasks, the vehicle is designed to provide a fairly good autonomy which is needed to face intermediate-range missions (100 nautical miles). The choice of a small waterplane area twin hull (SWATH) form has been motivated by its excellent properties of seakeeping qualities, combined with a non-conventional low resistance underwater hull shape, currently under patenting process, which is able to reduce to a minimum the resistance of the vessel especially at higher speeds. To obtain the most efficient profile of the underwater bodies, a systematic optimization with an automatic procedure based on a parametric definition of the geometry, a state-of-the-art computational fluid dynamics (CFD) flow solver, and a differential evolution global minimization algorithm have been created and used. As expected, all the final CFD computations on the best design have demonstrated the superior efficiency of the developed unconventional SWATH technology with respect to different alternatives of current hull typologies.

  16. Structural characterization of low level degradants in aztreonam injection and an innovative approach to aid HPLC method validation.

    PubMed

    Ye, Qingmei; Ding, Wei; Rinaldi, Frank; Huang, Yande; Miller, Scott A; Bolgar, Mark

    2016-05-30

    Three new degradants have been identified from drug product and active pharmaceutical ingredient stability samples of aztreonam, a marketed synthetic monocyclic beta-lactam antibiotic. The degradants were detected following the implementation of a new, more selective HPLC method for the determination of impurities and degradants. The new method was developed in response to changes in the regulatory requirement for mature products. Two of the new unknown Degradants (I and II) were observed in chromatograms from stability samples of aztreonam injection. The third new Degradant (III) was observed during a stability study of the aztreonam active pharmaceutical ingredient. These degradants were structurally characterized. A small amount (ca. 1-3mg) of each degradant was isolated via preparative HPLC for structure elucidation using accurate MS, one and two-dimensional NMR spectroscopy. The small amount of each NMR sample was then reused as a standard for HPLC purity/impurity method validation. Their exact concentrations were determined using quantitative NMR which enabled the execution of the quantitative elements of the HPLC method validation. This innovative approach eliminated the need to isolate or synthesize larger quantities of markers for HPLC/UV method validation, thus saving significant time and reducing costs. PMID:26991056

  17. EVALUATION OF THE CARBON FOOTPRINT OF AN INNOVATIVE SEWER REHABILITATION METHOD

    EPA Science Inventory

    A benefit of trenchless methods touted by many practitioners when compared to open cut construction is lower carbon dioxide emissions. In an attempt to verify these claims, tools have been developed that calculate the environmental impact of traditional open cut methods and commo...

  18. EVALUATION OF THE CARBON FOOTPRINT OF AN INNOVATIVE SEWER REHABILITATION METHOD - abstract

    EPA Science Inventory

    A benefit of trenchless methods touted by many practitioners when compared to open cut construction is lower carbon dioxide emissions. In an attempt to verify these claims, tools have been developed that calculate the environmental impact of traditional open cut methods and commo...

  19. The World Café: An Innovative Method to Facilitate Reflections on Internationalisation in Higher Education

    ERIC Educational Resources Information Center

    Estacio, Emee Vida; Karic, Toni

    2016-01-01

    This article provides an example of how the World Café method can be used as a platform to encourage reflection on internationalisation in higher education. The World Café is a community engagement method that encourages participants to engage in reflection and dialogue in a relaxed and comfortable atmosphere. It promotes the use of multiple…

  20. Development of an Innovative Algorithm for Aerodynamics-Structure Interaction Using Lattice Boltzmann Method

    NASA Technical Reports Server (NTRS)

    Mei, Ren-Wei; Shyy, Wei; Yu, Da-Zhi; Luo, Li-Shi; Rudy, David (Technical Monitor)

    2001-01-01

    The lattice Boltzmann equation (LBE) is a kinetic formulation which offers an alternative computational method capable of solving fluid dynamics for various systems. Major advantages of the method are owing to the fact that the solution for the particle distribution functions is explicit, easy to implement, and the algorithm is natural to parallelize. In this final report, we summarize the works accomplished in the past three years. Since most works have been published, the technical details can be found in the literature. Brief summary will be provided in this report. In this project, a second-order accurate treatment of boundary condition in the LBE method is developed for a curved boundary and tested successfully in various 2-D and 3-D configurations. To evaluate the aerodynamic force on a body in the context of LBE method, several force evaluation schemes have been investigated. A simple momentum exchange method is shown to give reliable and accurate values for the force on a body in both 2-D and 3-D cases. Various 3-D LBE models have been assessed in terms of efficiency, accuracy, and robustness. In general, accurate 3-D results can be obtained using LBE methods. The 3-D 19-bit model is found to be the best one among the 15-bit, 19-bit, and 27-bit LBE models. To achieve desired grid resolution and to accommodate the far field boundary conditions in aerodynamics computations, a multi-block LBE method is developed by dividing the flow field into various blocks each having constant lattice spacing. Substantial contribution to the LBE method is also made through the development of a new, generalized lattice Boltzmann equation constructed in the moment space in order to improve the computational stability, detailed theoretical analysis on the stability, dispersion, and dissipation characteristics of the LBE method, and computational studies of high Reynolds number flows with singular gradients. Finally, a finite difference-based lattice Boltzmann method is

  1. Cloning mice and men: prohibiting the use of iPS cells for human reproductive cloning.

    PubMed

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2010-01-01

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation.

  2. An Innovative Method for Rapid Identification and Detection of Vibrio alginolyticus in Different Infection Models

    PubMed Central

    Fu, Kaifei; Li, Jun; Wang, Yuxiao; Liu, Jianfei; Yan, He; Shi, Lei; Zhou, Lijun

    2016-01-01

    Vibrio alginolyticus is one of the most common pathogenic marine Vibrio species, and has been found to cause serious seafood-poisoning or fatal extra-intestinal infections in humans, such as necrotizing soft-tissue infections, bacteremia, septic shock, and multiple organ failures. Delayed accurate diagnosis and treatment of most Vibrio infections usually result to high mortality rates. The objective of this study was to establish a rapid diagnostic method to detect and identify the presence of V. alginolyticus in different samples, so as to facilitate timely treatment. The widely employed conventional methods for detection of V. alginolyticus include biochemical identification and a variety of PCR methods. The former is of low specificity and time-consuming (2–3 days), while the latter has improved accuracy and processing time. Despite such advancements, these methods are still complicated, time-consuming, expensive, require expertise and advanced laboratory systems, and are not optimal for field use. With the goal of providing a simple and efficient way to detect V. alginolyticus, we established a rapid diagnostic method based on loop-mediated Isothermal amplification (LAMP) technology that is feasible to use in both experimental and field environments. Three primer pairs targeting the toxR gene of V. alginolyticus were designed, and amplification was carried out in an ESE tube scanner and Real-Time PCR device. We successfully identified 93 V. alginolyticus strains from a total of 105 different bacterial isolates and confirmed their identity by 16s rDNA sequencing. We also applied this method on infected mouse blood and contaminated scallop samples, and accurate results were both easily and rapidly (20–60 min) obtained. Therefore, the RT-LAMP assay we developed can be conveniently used to detect the presence of V. alginolyticus in different samples. Furthermore, this method will also fulfill the gap for real-time screening of V. alginolyticus infections

  3. Review of established and innovative detection methods for carbapenemase-producing Gram-negative bacteria.

    PubMed

    Osei Sekyere, J; Govinden, U; Essack, S Y

    2015-11-01

    The minimal antibiotic options for carbapenemase-producing Gram-negative bacteria necessitate their rapid detection. A literature review of a variety of phenotypic and genotypic methods is presented. Advances in culture methods and screening media are still subject to long incubation hours. Biochemical methods have shorter turnaround times and higher sensitivities and specificities, but cannot differentiate between various types and variants. Spectrophotometric methods are cheap and efficient, but are uncommon in many clinical settings, while the MALDI-TOF MS is promising for species identification, typing and resistance gene determination. Although next generation sequencing (NGS) technologies provide a better platform to detect, type and characterize carbapenem-resistant bacteria, the different NGS platforms, the large computer memories and space needed to process and store genomic data and the nonuniformity in data analysis platforms are still a challenge. The sensitivities, specificities and turnaround times recorded in the various studies reviewed favours the use of the biochemical tests (Carba NP or Rapid Carb screen tests) for the detection of putative carbapenemase-producing isolates. MALDI-TOF MS and/or molecular methods like microarray, loop-mediated isothermal amplification and real-time multiplex PCR assays could be used for further characterization in a reference laboratory. NGS may be used for advanced epidemiological and molecular studies.

  4. An Innovative Method of Assessing the Mechanical Axis Deviation in the Lower Limb in Standing Position

    PubMed Central

    Kamath, Jagannath; Jayasheelan, Nikil; Singh, Rohit

    2016-01-01

    Introduction Various methods of measuring mechanical axis deviation of lower limb have been described including radiographic and CT scanogram, intraoperative fluoroscopy with the use of an electrocautery cord. These methods determine the mechanical axis in a supine, non-weight bearing position. Although long cassette standing radiographic view is used for the purpose but is not available at most centres. A dynamic method of determining the mechanical axis in a weight bearing position was devised in this study. Aim The aim of the study was to describe a simpler and newer method in quantifying the mechanical axis deviation in places where full length cassettes for standing X rays are not available. Materials and Methods A pilot study was conducted on 15 patients. The deviation from the mechanical axis was measured using a manually operated, hydraulic mechanism based, elevating scissor lift table. Patient was asked to stand erect over the elevating lift table with both patellae facing forward and C-arm image intensifier was positioned horizontally. Radiological markers were tied to a radio-opaque thread and placed at the centre of head of the femur and another at the centre of the tibio-talar joint. C-arm views of the hip, ankle and knee joint were taken to confirm the correct position of the marker by varying the height of the lift table. Results The mechanical axis deviation values were recorded by measuring distance between the centre of the knee and radio-opaque thread in cm. This was measured in each case both clinically and from the image on the monitor. The two values were found to be statistically same. Pain was measured on VAS. Mechanical axis deviation values and VAS score were found to be positively significantly correlated. Conclusion This technique is dynamic, unique and accurate as compared to other methods for assessing mechanical axis deviation in a weight bearing position. PMID:27504362

  5. An innovative exercise method to simulate orbital EVA work - Applications to PLSS automatic controls

    NASA Technical Reports Server (NTRS)

    Lantz, Renee; Vykukal, H.; Webbon, Bruce

    1987-01-01

    An exercise method has been proposed which may satisfy the current need for a laboratory simulation representative of muscular, cardiovascular, respiratory, and thermoregulatory responses to work during orbital extravehicular activity (EVA). The simulation incorporates arm crank ergometry with a unique body support mechanism that allows all body position stabilization forces to be reacted at the feet. By instituting this exercise method in laboratory experimentation, an advanced portable life support system (PLSS) thermoregulatory control system can be designed to more accurately reflect the specific work requirements of orbital EVA.

  6. Efficient computational methods to study new and innovative signal detection techniques in SETI

    NASA Technical Reports Server (NTRS)

    Deans, Stanley R.

    1991-01-01

    The purpose of the research reported here is to provide a rapid computational method for computing various statistical parameters associated with overlapped Hann spectra. These results are important for the Targeted Search part of the Search for ExtraTerrestrial Intelligence (SETI) Microwave Observing Project.

  7. Pedagogical Innovation and Music Education in Spain: Introducing the Dalcroze Method in Catalonia

    ERIC Educational Resources Information Center

    Comas Rubí, Francesca; Motilla-Salas, Xavier; Sureda-Garcia, Bernat

    2014-01-01

    The aim of this paper is to analyse how the Dalcroze method was introduced to Spain and became known there, more specifically in the Catalonia of the "Noucentisme" movement, and why it made the greatest impact and was more widely disseminated in this particular region of Spain. Following a summary of Dalcroze's contributions to…

  8. EVALUATION OF THE CARBON FOOTPRINT OF INNOVATIVE WATER MAIN REHABILITATION TECHNOLOGIES VS OPEN CUT METHODS

    EPA Science Inventory

    A major benefit of trenchless rehabilitation technologies touted by many practitioners when comparing their products with traditional open cut construction methods is lower carbon dioxide (CO2) emissions. In an attempt to verify these claims, multiple tools have been d...

  9. Innovative Solutions for Words with Emphasis: Alternative Methods of Braille Transcription

    ERIC Educational Resources Information Center

    Kamei-Hannan, Cheryl

    2009-01-01

    The author of this study proposed two alternative methods for transcribing words with emphasis into braille and compared the use of the symbols for emphasis with the current braille code. The results showed that students were faster at locating words presented in one of the alternate formats, but that there was no difference in students' accuracy…

  10. Citric acid facilitated thermal treatment: An innovative method for the remediation of mercury contaminated soil.

    PubMed

    Ma, Fujun; Peng, Changsheng; Hou, Deyi; Wu, Bin; Zhang, Qian; Li, Fasheng; Gu, Qingbao

    2015-12-30

    Thermal treatment is a promising technology for the remediation of mercury contaminated soils, but it often requires high energy input at heating temperatures above 600°C, and the treated soil is not suitable for agricultural reuse. The present study developed a novel method for the thermal treatment of mercury contaminated soils with the facilitation of citric acid (CA). A CA/Hg molar ratio of 15 was adopted as the optimum dosage. The mercury concentration in soils was successfully reduced from 134 mg/kg to 1.1mg/kg when treated at 400°C for 60 min and the treated soil retained most of its original soil physiochemical properties. During the treatment process, CA was found to provide an acidic environment which enhanced the volatilization of mercury. This method is expected to reduce energy input by 35% comparing to the traditional thermal treatment method, and lead to agricultural soil reuse, thus providing a greener and more sustainable remediation method for treating mercury contaminated soil in future engineering applications. PMID:26253234

  11. Innovative Mixed-Methods Research: Moving beyond Design Technicalities to Epistemological and Methodological Realizations

    ERIC Educational Resources Information Center

    Riazi, A. Mehdi

    2016-01-01

    Mixed-methods research (MMR), as an inter-discourse (quantitative and qualitative) methodology, can provide applied linguistics researchers the opportunity to draw on and integrate the strengths of the two research methodological approaches in favour of making more rigorous inferences about research problems. In this article, the argument is made…

  12. EVALUATION OF THE CARBON FOOTPRINT OF INNOVATIVE WATER MAIN REHABILITATION TECHNOLOGIES VS. OPEN CUT METHODS

    EPA Science Inventory

    A major benefit of trenchless rehabilitation technologies touted by many practitioners when comparing their products with tradition open cut construction methods is lower carbon dioxide (CO2) emissions. In an attempt to verify these claims, multiple tools have been dev...

  13. Medical innovation laws: an unnecessary innovation.

    PubMed

    Richards, Bernadette

    2016-06-01

    Objective This paper aims to demonstrate that any suggestion that there is a need for specific innovation laws is flawed. Innovation is central to good medical practice and is adequately supported by current law. Methods The paper reviews the nature of medical innovation and outlines recent attempts in the UK to introduce specific laws aimed at 'encouraging' and 'supporting' innovation. The current legal framework is outlined and the role of the law in relation to medical innovation explored. Results The analysis demonstrates the cyclic relationship between medical advancement and the law and concludes that there is no requirement for specific innovation laws. Conclusions The law not only supports innovation and development in medical treatment but encourages it as central to a functioning medical system. There is no need to introduce specific laws aimed at medical innovation; to do so represents an unnecessary legal innovation and serves to complicate matters. What is known about the topic? Over recent months, there has been a great deal of discussion surrounding the law in the context of medical innovation. This was driven by the attempts in the UK to introduce specific laws in the Medical Innovation Bill. The general subject matter - negligence and the expected standard of care in the provision of treatment - is very well understood, but not in cases where the treatment can be described as innovative. The general rhetoric in both the UK and Australia around the Medical Innovation Bill demonstrates a lack of understanding of the position of the law with regards to innovative treatment. What does this paper add? This paper adds clarity to the debate. It presents the law and explains the manner in which the law can operate around innovative treatment. The paper asserts that medical innovation is both supported and encouraged by existing legal principles. What are the implications for practitioners? The paper presents an argument that can guide the policy position

  14. DIFFUSION OF INNOVATION: ENHANCING THE DISSEMINATION OF THE PONSETI METHOD IN LATIN AMERICA THROUGH VIRTUAL FORUMS

    PubMed Central

    Jayawardena, Asitha; Boardman, Allison; Cook, Thomas; Oprescu, Florin; Morcuende, Jose A

    2011-01-01

    This ethnographic study evaluated the use of low-bandwidth web-conferencing to enhance diffusion of a specific best practice, the Ponseti method to treat clubfoot, in three economically diverse countries in Latin America. A “Ponseti Virtual Forum” (PVF) was organized in Guatemala, Peru and Chile to examine the influences of economic level and telecommunication infrastructure on the effectiveness of tins approach. Across the three countries, a total of 14 different sites participated in the PVFs. Thirty-three Ponseti-trained practitioners were interviewed before and after each PVF, which included interactions with a Spanish-speaking Ponseti method expert. Semi-structured interviews, observations, and IP address data were triangulated and analyzed. The results demonstrated that 100% of the practitioners rated the sessions as very useful and that they would use this approach again. The largest obstacles to using PVFs were financial (7 out of 9 practitioners) in Guatemala; a lack of equipment and network access (6 out of 11) in Peru; and the organization and implementation of the conferences themselves (7 out of 9) in Chile. This study illustrates the usefulness of Ponseti Virtual Forums in Latin America. Health officials in Peru are currently developing a large-scale information session for traumatologists about the Ponseti method, while practitioners in Guatemala and Chile are organizing monthly scholarly meetings for physicians in remote areas. This initial feedback suggests that low-bandwidth web-conferencing can be an important vehicle for the dissemination of best practices, such as the Ponseti method, in developing countries. PMID:22096417

  15. John Locke's "new method of making common-place-books": tradition, innovation and epistemic effects.

    PubMed

    Stolberg, Michael

    2014-01-01

    In 1676, the English physician and philosopher John Locke published a new method of commonplacing. He had developed this method and, in particular, a new approach to organizing and indexing the entries, in the course of 25 years of personal note-taking and it proved quite influential. This paper presents the three major approaches to commonplacing as practiced by physicians and other scholars before Locke--the systematic or textbook approach, the alphabetical approach and the sequential or index-based approach--and it analyzes the ways in which Locke himself applied them in his own commonplace books. In comparison with established approaches, his new method offered a maximum degree of flexibilitywhile facilitating the later retrieval of notes and minimising waste of space and paper. Thanks to these features, it was particularly well suited for physicians and natural philosophers who were interested in the infinite variety of natural particulars rather than in elegant quotes on a very limited set of classical topics. In conclusion, the potential epistemic impact of commonplacing on early modern medicine and natural philosophy is discussed, in particular its importance for contemporary debates about species and disease entities and for the emergence of the notion of "facts"!

  16. An Innovative Method to Involve Community Health Workers as Partners in Evaluation Research

    PubMed Central

    Issel, L. Michele; Townsell, Stephanie J.; Chapple-McGruder, Theresa; Handler, Arden

    2011-01-01

    Objectives. We developed a process through which community outreach workers, whose role is not typically that of a trained researcher, could actively participate in collection of qualitative evaluation data. Methods. Outreach workers for a community-based intervention project received training in qualitative research methodology and certification in research ethics. They used a Voice over Internet Protocol phone-in system to provide narrative reports about challenges faced by women they encountered in their outreach activities as well as their own experiences as outreach workers. Results. Qualitative data contributed by outreach workers provided insights not otherwise available to the evaluation team, including details about the complex lives of underserved women at risk for poor pregnancy outcomes and the challenges and rewards of the outreach worker role. Conclusions. Lay health workers can be a valuable asset as part of a research team. Training in research ethics and methods can be tailored to their educational level and preferences, and their insights provide important information and perspectives that may not be accessible via other data collection methods. Challenges encountered in the dual roles of researcher and lay health worker can be addressed in training. PMID:22021290

  17. Closed vessel microwave assisted extraction - An innovative method for determination of trace metals in plant materials

    NASA Astrophysics Data System (ADS)

    Oeztan, S.; Duering, R.-A.

    2012-04-01

    Determination of metal concentrations in plant samples is important for better understanding of effects of toxic metals that are biologically magnified through the food chain and compose a great danger to all living beings. In recent years the use of microwave assisted extraction for plant samples has shown tremendous research interest which will probably substitute conventional procedures in the future. Generally conventional procedures have disadvantages including consuming of time and solvents. The objective of this study is to investigate and compare a new closed vessel microwave extraction (MAE) method with the combination of EDTA (MAE-EDTA) for the determination of metal contents (Cd, Mn, Pb, Zn) in plant samples (Lolio-Cynosuretum) by ICP-OES. Validation of the method was done by comparison of the results with another MAE procedure (MAE-H) which is applied with the mixture of 69% nitric acid (HNO3) and 30% hydrogen peroxide (H2O2). Moreover, conventional plant extraction (CE) method, for which the dissolution of plant samples were handled in HNO3 after dry ashing at 420° C, was used as a reference method. Approximately 0.5 g of sample was digested in 5 ml HNO3, 3 ml H2O2, and 5 ml deionized H2O for MAE-H and in 8 ml EDTA solution for MAE-EDTA. Certified plant reference materials (CRMs) were used for comparison of recovery rates from different extraction protocols. Thereby, the applicability of both MAE-H and MAE-EDTA procedures could be demonstrated. For 58 plant samples MAE-H showed the same extraction yields as CE in the determination of trace metal contents of the investigated elements in plant samples. MAE-EDTA gave similar values when compared to MAE-H and highly linear relationships were found for determination of Cd, Mn, Pb and Zn amounts. The recoveries for the CRMs were within the range 89.6-115%. Finally, strategic characteristics of MAE-EDTA for determination metal contents (Cd, Mn, Pb, Zn) in plant samples are: (i) applicability to a large set

  18. Investigation of an innovative method for DC flow suppression of double-inlet pulse tube coolers

    NASA Astrophysics Data System (ADS)

    Hu, J. Y.; Luo, E. C.; Wu, Z. H.; Dai, W.; Zhu, S. L.

    2007-05-01

    The use of double-inlet mode in the pulse tube cooler opens up a possibility of DC flow circulating around the regenerator and the pulse tube. The DC flow sometimes deteriorates the performance of the cryocooler because such a steady flow adds an unwanted thermal load to the cold heat exchanger. It seems that this problem is still not well solved although a lot of effort has been made. Here we introduce a membrane-barrier method for DC flow suppression in double-inlet pulse tube coolers. An elastic membrane is installed between the pulse tube cooler inlet and the double-inlet valve to break the closed-loop flow path of DC flow. The membrane is acoustically transparent, but would block the DC flow completely. Thus the DC flow is thoroughly suppressed and the merit of double-inlet mode is remained. With this method, a temperature reduction of tens of Kelvin was obtained in our single-stage pulse tube cooler and the lowest temperature reached 29.8 K.

  19. On cloning human beings.

    PubMed

    de Melo-Martin, Inmaculada

    2002-06-01

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

  20. Innovative self-calibration method for accelerometer scale factor of the missile-borne RINS with fiber optic gyro.

    PubMed

    Zhang, Qian; Wang, Lei; Liu, Zengjun; Zhang, Yiming

    2016-09-19

    The calibration of an inertial measurement unit (IMU) is a key technique to improve the preciseness of the inertial navigation system (INS) for missile, especially for the calibration of accelerometer scale factor. Traditional calibration method is generally based on the high accuracy turntable, however, it leads to expensive costs and the calibration results are not suitable to the actual operating environment. In the wake of developments in multi-axis rotational INS (RINS) with optical inertial sensors, self-calibration is utilized as an effective way to calibrate IMU on missile and the calibration results are more accurate in practical application. However, the introduction of multi-axis RINS causes additional calibration errors, including non-orthogonality errors of mechanical processing and non-horizontal errors of operating environment, it means that the multi-axis gimbals could not be regarded as a high accuracy turntable. As for its application on missiles, in this paper, after analyzing the relationship between the calibration error of accelerometer scale factor and non-orthogonality and non-horizontal angles, an innovative calibration procedure using the signals of fiber optic gyro and photoelectric encoder is proposed. The laboratory and vehicle experiment results validate the theory and prove that the proposed method relaxes the orthogonality requirement of rotation axes and eliminates the strict application condition of the system. PMID:27661867

  1. Technologic Advances in Aural Rehabilitation: Applications and Innovative Methods of Service Delivery

    PubMed Central

    Sweetow, Robert W.; Sabes, Jennifer Henderson

    2007-01-01

    The level of interest in aural rehabilitation has increased recently, both in clinical use and in research presentations and publications. Advances in aural rehabilitation have seen previous techniques such as speech tracking and analytic auditory training reappear in computerized forms. These new delivery methods allow for a consistent, cost-effective, and convenient training program. Several computerized aural rehabilitation programs for hearing aid wearers and cochlear implant recipients have recently been developed and were reported on at the 2006 State of the Science Conference of the Rehabilitation Engineering Research Center on Hearing Enhancement at Gallaudet University. This article reviews these programs and outlines the similarities and differences in their design. Another promising area of aural rehabilitation research is the use of pharmaceuticals in the rehabilitation process. The results from a study of the effect of d-amphetamine in conjunction with intensive aural rehabilitation with cochlear implant patients are also described. PMID:17494876

  2. Innovative Adaptive Control Method Demonstrated for Active Suppression of Instabilities in Engine Combustors

    NASA Technical Reports Server (NTRS)

    Kopasakis, George

    2005-01-01

    This year, an improved adaptive-feedback control method was demonstrated that suppresses thermoacoustic instabilities in a liquid-fueled combustor of a type used in aircraft engines. Extensive research has been done to develop lean-burning (low fuel-to-air ratio) combustors that can reduce emissions throughout the mission cycle to reduce the environmental impact of aerospace propulsion systems. However, these lean-burning combustors are susceptible to thermoacoustic instabilities (high-frequency pressure waves), which can fatigue combustor components and even downstream turbine blades. This can significantly decrease the safe operating life of the combustor and turbine. Thus, suppressing the thermoacoustic combustor instabilities is an enabling technology for meeting the low-emission goals of the NASA Ultra-Efficient Engine Technology (UEET) Project.

  3. Innovative computer-aided methods for the discovery of new kinase ligands.

    PubMed

    Abuhammad, Areej; Taha, Mutasem

    2016-04-01

    Recent evidence points to significant roles played by protein kinases in cell signaling and cellular proliferation. Faulty protein kinases are involved in cancer, diabetes and chronic inflammation. Efforts are continuously carried out to discover new inhibitors for selected protein kinases. In this review, we discuss two new computer-aided methodologies we developed to mine virtual databases for new bioactive compounds. One method is ligand-based exploration of the pharmacophoric space of inhibitors of any particular biotarget followed by quantitative structure-activity relationship-based selection of the best pharmacophore(s). The second approach is structure-based assuming that potent ligands come into contact with binding site spots distinct from those contacted by weakly potent ligands. Both approaches yield pharmacophores useful as 3D search queries for the discovery of new bioactive (kinase) inhibitors. PMID:27105126

  4. Innovative educational methods and technologies applicable to continuing professional development in periodontology.

    PubMed

    Mattheos, N; Schoonheim-Klein, M; Walmsley, A D; Chapple, I L C

    2010-05-01

    Continuous professional development (CPD) in Periodontology refers to the overall framework of opportunities that facilitate a life-long learning practice, driven by the learner-practitioner and supported by a variety of institutions and individuals. CPD must address different needs for a great diversity of practitioners. It is clear that no particular methodology or technology is able to successfully accommodate the entire spectrum of CPD in Periodontology. Course designers must choose from and combine a wide array of methodologies and technologies, depending upon the needs of the learners and the objectives of the intended education. Research suggests that 'interactivity', 'flexibility', 'continuity' and 'relevance to learners' practice' are major characteristics of successful CPD. Various methods of mentoring, peer-learning environments and work-based learning have been combined with reflective practice and self-study to form the methodological backbone of CPD courses. Blended learning encompasses a wide array of technologies and methodologies and has been successfully used in CPD courses. Internet-based content learning management systems, portable Internet devices, powerful databases and search engines, together with initiatives such as 'open access' and 'open courseware' provide an array of effective instructional and communication tools. Assessment remains a key issue in CPD, providing learners with valuable feedback and it ensures the credibility and effectiveness of the learning process. Assessment is a multi-level process using different methods for different learning outcomes, as directed by current evidence and best practices. Finally, quality assurance of the education provided must follow CPD courses at all times through a structured and credible process. PMID:20415976

  5. New methods as alternative or corrective measures for the pitfalls and artifacts of reverse transcription and polymerase chain reactions (RT-PCR) in cloning chimeric or antisense-accompanied RNA.

    PubMed

    Yuan, Chengfu; Liu, Yongming; Yang, Min; Liao, D Joshua

    2013-06-01

    We established new methods for cloning cDNA ends that start with reverse transcription (RT) and soon proceed with the synthesis of the second cDNA strand, avoiding manipulations of fragile RNA. Our 3'-end cloning method does not involve poly-dT primers and polymerase chain reactions (PCR), is low in efficiency but high in fidelity and can clone those RNAs without a poly-A tail. We also established a cDNA protection assay to supersede RNA protection assay. The protected cDNA can be amplified, cloned and sequenced, enhancing sensitivity and fidelity. We report that RT product using gene-specific primer (GSP) cannot be gene- or strand-specific because RNA sample contains endogenous random primers (ERP). The gene-specificity may be improved by adding a linker sequence at the 5'-end of the GSP to prime RT and using the linker as a primer in the ensuing PCR. The strand-specificity may be improved by using strand-specific DNA oligos in our protection assay. The CDK4 mRNA and TSPAN31 mRNA are transcribed from the opposite DNA strands and overlap at their 3' ends. Using this relationship as a model, we found that the overlapped sequence might serve as a primer with its antisense as the template to create a wrong-template extension in RT or PCR. We infer that two unrelated RNAs or cDNAs overlapping at the 5'- or 3'-end might create a spurious chimera in this way, and many chimeras with a homologous sequence may be such artifacts. The ERP and overlapping antisense together set complex pitfalls, which one should be aware of.

  6. Realization of a universal and phase-covariant quantum cloning machine in separate cavities

    SciTech Connect

    Fang Baolong; Song Qingming; Ye Liu

    2011-04-15

    We present a scheme to realize a special quantum cloning machine in separate cavities. The quantum cloning machine can copy the quantum information from a photon pulse to two distant atoms. Choosing the different parameters, the method can perform optimal symmetric (asymmetric) universal quantum cloning and optimal symmetric (asymmetric) phase-covariant cloning.

  7. Innovative method for recovery and valorization of hydroxytyrosol from olive mill wastewaters.

    PubMed

    Bonetti, A; Venturini, S; Ena, A; Faraloni, C

    2016-01-01

    The nutritional properties of olive oil can be attributed to its oleic acid and phenolic compounds content, acting as natural oxidants to prevent human diseases. In particular, hydroxytyrosol has an anti-inflammatory action similar to omega 3 fatty acids from fish oil. The olive oil production was conducted by two extraction procedures: first, a two-phase extraction giving extra-virgin olive oil and humid pomace, second, a three-phase working process of humid pomace, obtaining another minimum quantity of extra-virgin olive oil, 'dry' pomace devoid of polyphenols, and mill wastewaters rich in anti-oxidant compounds. The aim of this processing was to employ water to extract the highest concentration of polyphenols from humid pomace and convey them in oil mill wastewaters for extraction. Processed olives were 37,200 kg, pomace deprived of polyphenols was equal to 20,400 kg and processing was performed with 500 kg of olives per hour. This method offers advantages of using cheap equipment and technical simplicity. PMID:27386985

  8. Improved dengue fever prevention through innovative intervention methods in the city of Salto, Uruguay

    PubMed Central

    Basso, César; García da Rosa, Elsa; Romero, Sonnia; González, Cristina; Lairihoy, Rosario; Roche, Ingrid; Caffera, Ruben M.; da Rosa, Ricardo; Calfani, Marisel; Alfonso-Sierra, Eduardo; Petzold, Max; Kroeger, Axel; Sommerfeld, Johannes

    2015-01-01

    Background Uruguay is located at the southern border of Aedes aegypti distribution on the South American sub-continent. The reported dengue cases in the country are all imported from surrounding countries. One of the cities at higher risk of local dengue transmission is Salto, a border city with heavy traffic from dengue endemic areas. Methods We completed an intervention study using a cluster randomized trial design in 20 randomly selected ‘clusters’ in Salto. The clusters were located in neighborhoods of differing geography and economic, cultural and social aspects. Results Entomological surveys were carried out to measure the impact of the intervention on vector densities. Through participatory processes of all stakeholders, an appropriate ecosystem management intervention was defined. Residents collected the abundant small water holding containers and the Ministry of Public Health and the Municipality of Salto were responsible for collecting and eliminating them. Additional vector breeding places were large water tanks; they were either altered so that they could not hold water any more or covered so that oviposition by mosquitoes could not take place. Conclusions The response from the community and national programme managers was encouraging. The intervention evidenced opportunities for cost savings and reducing dengue vector densities (although not to statistically significant levels). The observed low vector density limits the potential reduction due to the intervention. A larger sample size is needed to obtain a statistically significant difference. PMID:25604764

  9. Cement Calcaneoplasty: An Innovative Method for Treating Nonunion in Calcaneal Insufficiency Fracture.

    PubMed

    Godavitarne, Charles; Fawzy, Ernest; Giancola, Giorgio; Louette, Luc

    2016-01-01

    Insufficiency type stress fractures are common in older patients with osteoporosis. Persistent pain after nonunion of these fractures can be disabling, with the management options often limited. We aimed to assess the suitability of fluoroscopic-guided injection of bone cement into a persistently symptomatic nonuniting calcaneal insufficiency fracture. To the best of our knowledge, this technique has not previously been described in the published data. After local subcutaneous anesthesia, the midpoint of the fracture site was accessed by trocar insertion under radiographic guidance, and bone cement was injected directly into the site. A preprocedure visual analog scale pain score of 90 of 100 was recorded. This had improved to 0 of 100 at the 12-month follow-up point after the procedure. The aim of the present case report was to raise awareness of percutaneous calcaneoplasty, which we believe to be a safe and well-tolerated technique for the management of osteoporotic insufficiency fracture of the calcaneus. We propose that this technique be considered when conservative methods aimed at promoting fracture healing have failed. PMID:26875768

  10. Cloning in America: constitutional rights and limits.

    PubMed

    Erwin, C

    2000-01-01

    As readers of science fiction are well aware, the term "clone" refers to asexually produced offspring, that is, produced by a process of cell-division which does not begin with the union of two sex cells. A clone would be the genetic twin of the cell donor. Propagation of plants by this method is, of course, commonplace, but mammalian reproduction in this fashion would be indeed a revolutionary accomplishment, with profound and disturbing implications. PMID:14627028

  11. "The Sweetness of Struggle": Innovation in Physical Education Teacher Education through "Student-Centered Inquiry as Curriculum" in a Physical Education Methods Course

    ERIC Educational Resources Information Center

    Oliver, Kimberly L.; Oesterreich, Heather A.; Aranda, Raquel; Archeleta, Jarrod; Blazer, Casey; de la Cruz, Kandy; Martinez, Daniel; McConnell, Jenn; Osta, Maggee; Parks, Lacie; Robinson, Rinalldo

    2015-01-01

    Purpose: The purpose of this study was to examine the challenges and benefits that emerged while using an innovative field-based student-centered inquiry as curriculum model in a secondary physical education methods course. Participants and setting: This study took place in the Southwest USA. Participants included 11 pre-service teachers who were…

  12. An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae.

    PubMed

    Wang, Sheng; Xing, Haiying; Hua, Chenlei; Guo, Hui-Shan; Zhang, Jie

    2016-06-01

    The soilborne fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)-based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae. PMID:26780432

  13. An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae.

    PubMed

    Wang, Sheng; Xing, Haiying; Hua, Chenlei; Guo, Hui-Shan; Zhang, Jie

    2016-06-01

    The soilborne fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)-based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae.

  14. Do Managers Clone Themselves?

    ERIC Educational Resources Information Center

    Baron, Alma S.

    1981-01-01

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

  15. Statement on Human Cloning

    MedlinePlus

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

  16. Twins: A cloning experience.

    PubMed

    Prainsack, Barbara; Spector, Tim D

    2006-11-01

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

  17. Transposon-containing DNA cloning vector and uses thereof

    DOEpatents

    Berg, Claire M.; Berg, Douglas E.; Wang, Gan

    1997-01-01

    The present invention discloses a rapid method of restriction mapping, sequencing or localizing genetic features in a segment of deoxyribonucleic acid (DNA) that is up to 42 kb in size. The method in part comprises cloning of the DNA segment in a specialized cloning vector and then isolating nested deletions in either direction in vivo by intramolecular transposition into the cloned DNA. A plasmid has been prepared and disclosed.

  18. Transposon-containing DNA cloning vector and uses thereof

    DOEpatents

    Berg, C.M.; Berg, D.E.; Wang, G.

    1997-07-08

    The present invention discloses a rapid method of restriction mapping, sequencing or localizing genetic features in a segment of deoxyribonucleic acid (DNA) that is up to 42 kb in size. The method in part comprises cloning of the DNA segment in a specialized cloning vector and then isolating nested deletions in either direction in vivo by intramolecular transposition into the cloned DNA. A plasmid has been prepared and disclosed. 4 figs.

  19. Impermanence of bacterial clones

    PubMed Central

    Bobay, Louis-Marie; Traverse, Charles C.; Ochman, Howard

    2015-01-01

    Bacteria reproduce asexually and pass on a single genome copied from the parent, a reproductive mode that assures the clonal descent of progeny; however, a truly clonal bacterial species is extremely rare. The signal of clonality can be interrupted by gene uptake and exchange, initiating homologous recombination that results in the unique sequence of one clone being incorporated into another. Because recombination occurs sporadically and on local scales, these events are often difficult to recognize, even when considering large samples of completely sequenced genomes. Moreover, several processes can produce the appearance of clonality in populations that undergo frequent recombination. The rates and consequences of recombination have been studied in Escherichia coli for over 40 y, and, during this time, there have been several shifting views of its clonal status, population structure, and rates of gene exchange. We reexamine the studies and retrace the evolution of the methods that have assessed the extent of DNA flux, largely focusing on its impact on the E. coli genome. PMID:26195749

  20. Exponential megapriming PCR (EMP) cloning--seamless DNA insertion into any target plasmid without sequence constraints.

    PubMed

    Ulrich, Alexander; Andersen, Kasper R; Schwartz, Thomas U

    2012-01-01

    We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP further enables simultaneous cloning of multiple inserts.

  1. A novel method for the production of transgenic cloned pigs: electroporation-mediated gene transfer to non-cultured cells and subsequent selection with puromycin.

    PubMed

    Watanabe, Satoshi; Iwamoto, Masaki; Suzuki, Shun-ichi; Fuchimoto, Daiichiro; Honma, Daisuke; Nagai, Takashi; Hashimoto, Michiko; Yazaki, Satoko; Sato, Masahiro; Onishi, Akira

    2005-02-01

    Puromycin N-acetyl transferase gene (pac), of which the gene product catalyzes antibiotic puromycin (an effective inhibitor of protein synthesis), has been widely used as a dominant selection marker in embryonic stem (ES) cell-mediated transgenesis. The present study is the first to report on the usefulness of puromycin for production of enhanced green fluorescent protein (EGFP) transgenic piglets after somatic cell cloning and embryo transfer. Somatic cells isolated from porcine fetuses at 73 days of gestation were immediately electroporated with a transgene (pCAG-EGFPac) carrying both EGFP cDNA and pac. This procedure aims to avoid aging effects thought to be generated during cell culture. The recombinant cells were selected with puromycin at a low concentration (2 microg/ml), cultured for 7 days, and then screened for EGFP expression before somatic cell cloning. The manipulated embryos were transplanted into the oviducts of 14 foster mother sows. Four of the foster sows became pregnant and nine piglets were delivered. Of the nine piglets, eight died shortly after birth and one grew healthy after weaning. Results indicate that puromycin can be used for the selection of recombinant cells from noncultured cells, and moreover, may confer the production of genetically engineered newborns via nuclear transfer techniques in pigs. PMID:15385422

  2. Innovative non-destructive evaluation methods on HTR fuel at AREVA NP: towards a 100% non invasive control strategy

    SciTech Connect

    Banchet, J.; Tisseur, D.; Hermosilla Lara, S.; Piriou, M.; Bargain, R.; Guillermier, P.

    2007-07-01

    High Temperature Reactor (HTR) fuel consists in millimetric multilayered particles called TRISO, embedded, depending on the reactor design, in a pebble or cylinder-shaped graphite matrix called compact. Particles are typically composed of a 500 {mu}m fissile material kernel, a 95 {mu}m porous carbon layer called buffer, a 40 {mu}m dense pyrolytic carbon layer, a 35 {mu}m silicon carbide layer and another 40 {mu}m dense pyrolytic carbon layer. In order to ensure fuel qualification, as well as reactor safety, particles and compacts need to satisfy specifications concerning their physical characteristics and their integrity. In particular, geometrical parameters such as particle diameter and sphericity as well as layers thickness, but also layers density and the absence of structural defects such as cracks or de-cohesions need to be detected and characterized. In the past, a huge R and D work was carried out to build a TRISO particle characterization quality control plan, mainly based on particle sampling as well as destructive characterization methods. However, since then, development of industrial non-destructive evaluation techniques and devices contributed to envisage not only a non invasive control of HTR fuel, but also a 100% production control strategy. Since 2004, AREVA NP is engaged in a R and D program aiming at the development of innovative industrial nondestructive evaluation methods for HTR fuel. After investigating a number of potential techniques, some of them were selected based on their performances and/or their industrial potential. In particular, development has been carried out on high resolution X-Ray imaging allowing accurate layer thickness, layer density and structural defects characterization, X-Ray tomography offering the possibility to characterize fuel element homogeneity and determine the number of in-contact particles contained in a fuel element, infrared thermal imaging (ITI) allowing cracks detection, eddy currents (EC) enabling

  3. Innovative Universities.

    ERIC Educational Resources Information Center

    Barsi, Louis M.; Kaebnick, Gweneth W.

    1989-01-01

    The phenomenon of innovation within the university is examined, noting the possibility of innovation as a key to college vitality. A study was conducted using a group of institutions that demonstrated recent innovative spirit. Members of the American Association of State Colleges and Universities (AASCU), each has been recognized in an annual…

  4. Encouraging innovation.

    PubMed

    Hyman, Anthony A

    2014-02-01

    Innovation is central to the scientific endeavor, and yet the current system of funding in the United States discourages innovation, especially in the young. Subtle alterations to the funding system, guided in part by the success of the European Research Council, could have major effects on encouraging innovation.

  5. Developing Communities of Innovation by Identifying Innovation Champions

    ERIC Educational Resources Information Center

    Coakes, Elayne; Smith, Peter

    2007-01-01

    Purpose: The purpose of this paper is to propose that a form of communities of practice (CoP), a community of innovation (CoInv), is the best support for sustainable innovation. It aims to outline a method for identifying champions of innovation in organisation. Design/methodology/approach: The paper draws on extant research to argue that…

  6. Extremal quantum cloning machines

    SciTech Connect

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

    2005-10-15

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

  7. Cloning the laboratory mouse.

    PubMed

    Wakayama, T; Yanagimachi, R

    1999-06-01

    A brief account is given of early attempts to clone mammals (mice) by transferring cells (nuclei) of preimplantation embryos into enucleated oocytes, zygotes or blastomeres of two-cell embryos. This is followed by a brief review of recent successes using adult somatic cells: mammary gland cells for sheep, muscle cells for cattle and cumulus cells for mice. We have developed a technique for cloning the laboratory mouse by transferring cumulus cell nuclei into enucleated oocytes. With this technique, we have produced a population of over 80 cloned animals, and have carried the process over four generations. Development and fertility of these appear normal. However, the yield is very low; only approximately 1% of injected oocytes are carried to term. The challenge is now to understand the reason for this high loss. Is it a problem of technique, genomic reprogramming, somatic mutation, imprinting or incompatible cell cycle phases?

  8. Innovation network

    PubMed Central

    Acemoglu, Daron; Akcigit, Ufuk; Kerr, William R.

    2016-01-01

    Technological progress builds upon itself, with the expansion of invention in one domain propelling future work in linked fields. Our analysis uses 1.8 million US patents and their citation properties to map the innovation network and its strength. Past innovation network structures are calculated using citation patterns across technology classes during 1975–1994. The interaction of this preexisting network structure with patent growth in upstream technology fields has strong predictive power on future innovation after 1995. This pattern is consistent with the idea that when there is more past upstream innovation for a particular technology class to build on, then that technology class innovates more. PMID:27681628

  9. Human cloning: category, dignity, and the role of bioethics.

    PubMed

    Shuster, Evelyne

    2003-10-01

    Human cloning has been simultaneously a running joke for massive worldwide publicity of fringe groups like the Raelians, and the core issue of an international movement at the United Nations in support of a treaty to ban the use of cloning techniques to produce a child (so called reproductive cloning). Yet, even though debates on human cloning have greatly increased since the birth of Dolly, the clone sheep, in 1997, we continue to wonder whether cloning is after all any different from other methods of medically assisted reproduction, and what exactly makes cloning an 'affront to the dignity of humans.' Categories we adopt matter mightily as they inform but can also misinform and lead to mistaken and unproductive decisions. And thus bioethicists have a responsibility to ensure that the proper categories are used in the cloning debates and denounce those who try to win the ethical debate through well-crafted labels rather than well-reasoned argumentations. But it is as important for bioethicists to take a position on broad issues such as human cloning and species altering interventions. One 'natural question' would be, for example, should there be an international treaty to ban human reproductive cloning?

  10. Human cloning: category, dignity, and the role of bioethics.

    PubMed

    Shuster, Evelyne

    2003-10-01

    Human cloning has been simultaneously a running joke for massive worldwide publicity of fringe groups like the Raelians, and the core issue of an international movement at the United Nations in support of a treaty to ban the use of cloning techniques to produce a child (so called reproductive cloning). Yet, even though debates on human cloning have greatly increased since the birth of Dolly, the clone sheep, in 1997, we continue to wonder whether cloning is after all any different from other methods of medically assisted reproduction, and what exactly makes cloning an 'affront to the dignity of humans.' Categories we adopt matter mightily as they inform but can also misinform and lead to mistaken and unproductive decisions. And thus bioethicists have a responsibility to ensure that the proper categories are used in the cloning debates and denounce those who try to win the ethical debate through well-crafted labels rather than well-reasoned argumentations. But it is as important for bioethicists to take a position on broad issues such as human cloning and species altering interventions. One 'natural question' would be, for example, should there be an international treaty to ban human reproductive cloning? PMID:14959720

  11. PCR Duplication: A One-Step Cloning-Free Method to Generate Duplicated Chromosomal Loci and Interference-Free Expression Reporters in Yeast

    PubMed Central

    Huber, Florian; Meurer, Matthias; Bunina, Daria; Kats, Ilia; Maeder, Céline I.; Štefl, Martin; Mongis, Cyril; Knop, Michael

    2014-01-01

    Here, we report on a novel PCR targeting-based strategy called ‘PCR duplication’ that enables targeted duplications of genomic regions in the yeast genome using a simple PCR-based approach. To demonstrate its application we first duplicated the promoter of the FAR1 gene in yeast and simultaneously inserted a GFP downstream of it. This created a reporter for promoter activity while leaving the FAR1 gene fully intact. In another experiment, we used PCR duplication to increase the dosage of a gene in a discrete manner, from 1× to 2x. Using TUB4, the gene encoding for the yeast γ-tubulin, we validated that this led to corresponding increases in the levels of mRNA and protein. PCR duplication is an easy one-step procedure that can be adapted in different ways to permit rapid, disturbance-free investigation of various genomic regulatory elements without the need for ex vivo cloning. PMID:25493941

  12. Organizational Agility and Complex Enterprise System Innovations: A Mixed Methods Study of the Effects of Enterprise Systems on Organizational Agility

    ERIC Educational Resources Information Center

    Kharabe, Amol T.

    2012-01-01

    Over the last two decades, firms have operated in "increasingly" accelerated "high-velocity" dynamic markets, which require them to become "agile." During the same time frame, firms have increasingly deployed complex enterprise systems--large-scale packaged software "innovations" that integrate and automate…

  13. A method for the identification of promoters recognized by RNA polymerase containing a particular sigma factor: cloning of a developmentally regulated promoter and corresponding gene directed by the Streptomyces aureofaciens sigma factor RpoZ.

    PubMed

    Nováková, R; Sevcíková, B; Kormanec, J

    1998-02-16

    We have developed a method for the identification of promoters recognized by a particular sigma factor of RNA polymerase, based on a two-compatible plasmid system in Escherichia coli (Ec). Using the method, a DNA fragment containing the promoter, PREN40, recognized by sporulation-specific Streptomyces aureofaciens (Sa) sigma factor RpoZ, was cloned. High-resolution S1 nuclease mapping using RNA prepared from Ec, and Sa from various developmental stages has shown a high degree of similarity of PREN40 to consensus sequence of flagellar and chemotaxis promoters. The promoter was induced at the time of aerial mycelium formation, and was off in the Sa strain with the rpoZ-disrupted gene. A promoter-bearing DNA fragment was inserted into the promoter-probe plasmid pARC1 to give expression patterns consistent with the results of direct RNA analysis. The region downstream of the promoter was cloned in Sa. Sequence analysis revealed an open reading frame (ORF) of 283 amino acids (Mr 30006), encoding a highly basic (pI 12.35) protein with high percentage of serine, threonine and alanine (41.8%). PMID:9479043

  14. Applications of quantum cloning

    NASA Astrophysics Data System (ADS)

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

    2011-10-01

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

  15. The Cloning of America.

    ERIC Educational Resources Information Center

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

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

  16. Human therapeutic cloning.

    PubMed

    Lanza, R P; Cibelli, J B; West, M D

    1999-09-01

    Somatic cell nuclear 'reprogramming' in livestock species is now routine in many laboratories. Here, Robert Lanza, Jose Cibelli and Michael West discuss how these techniques may soon be used to clone genetically matched cells and tissues for transplantation into patients suffering from a wide range of disorders that result from tissue loss or dysfunction.

  17. Detectability of Plasmodium falciparum clones

    PubMed Central

    2010-01-01

    Background In areas of high transmission people often harbour multiple clones of Plasmodium falciparum, but even PCR-based diagnostic methods can only detect a fraction (the detectability, q) of all clones present in a host. Accurate measurements of detectability are desirable since it affects estimates of multiplicity of infection, prevalence, and frequency of breakthrough infections in clinical drug trials. Detectability can be estimated by typing repeated samples from the same host but it has been unclear what should be the time interval between the samples and how the data should be analysed. Methods A longitudinal molecular study was conducted in the Kassena-Nankana district in northern Ghana. From each of the 80 participants, four finger prick samples were collected over a period of 8 days, and tested for presence of different Merozoite Surface Protein (msp) 2 genotypes. Implications for estimating q were derived from these data by comparing the fit of statistical models of serial dependence and over-dispersion. Results The distribution of the frequencies of detection for msp2 genotypes was close to binomial if the time span between consecutive blood samples was at least 7 days. For shorter intervals the probabilities of detection were positively correlated, i.e. the shorter the interval between two blood collections, the more likely the diagnostic results matched for a particular genotype. Estimates of q were rather insensitive to the statistical model fitted. Conclusions A simple algorithm based on analysing blood samples collected 7 days apart is justified for generating robust estimates of detectability. The finding of positive correlation of detection probabilities for short time intervals argues against imperfect detection being directly linked to the 48-hour periodicity of P. falciparum. The results suggest that the detectability of a given parasite clone changes over time, at an unknown rate, but fast enough to regard blood samples taken one week

  18. [Nuclear transfer and therapeutic cloning].

    PubMed

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

    2005-03-01

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

  19. The First Human Cloned Embryo.

    ERIC Educational Resources Information Center

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

    2002-01-01

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

  20. Cloning of Plasmodium falciparum by single-cell sorting.

    PubMed

    Miao, Jun; Li, Xiaolian; Cui, Liwang

    2010-10-01

    Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two-dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples.

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

    PubMed Central

    Fowler, Daniel K.; Stewart, Scott; Seredick, Steve; Eisen, Judith S.

    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. PMID:27500400

  2. Probabilistic Cloning and Quantum Computation

    NASA Astrophysics Data System (ADS)

    Gao, Ting; Yan, Feng-Li; Wang, Zhi-Xi

    2004-06-01

    We discuss the usefulness of quantum cloning and present examples of quantum computation tasks for which the cloning offers an advantage which cannot be matched by any approach that does not resort to quantum cloning. In these quantum computations, we need to distribute quantum information contained in the states about which we have some partial information. To perform quantum computations, we use a state-dependent probabilistic quantum cloning procedure to distribute quantum information in the middle of a quantum computation.

  3. IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly.

    PubMed

    García-Nafría, Javier; Watson, Jake F; Greger, Ingo H

    2016-06-06

    In vivo homologous recombination holds the potential for optimal molecular cloning, however, current strategies require specialised bacterial strains or laborious protocols. Here, we exploit a recA-independent recombination pathway, present in widespread laboratory E.coli strains, to develop IVA (In Vivo Assembly) cloning. This system eliminates the need for enzymatic assembly and reduces all molecular cloning procedures to a single-tube, single-step PCR, performed in <2 hours from setup to transformation. Unlike other methods, IVA is a complete system, and offers significant advantages over alternative methods for all cloning procedures (insertions, deletions, site-directed mutagenesis and sub-cloning). Significantly, IVA allows unprecedented simplification of complex cloning procedures: five simultaneous modifications of any kind, multi-fragment assembly and library construction are performed in approximately half the time of current protocols, still in a single-step fashion. This system is efficient, seamless and sequence-independent, and requires no special kits, enzymes or proprietary bacteria, which will allow its immediate adoption by the academic and industrial molecular biology community.

  4. IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly

    PubMed Central

    García-Nafría, Javier; Watson, Jake F.; Greger, Ingo H.

    2016-01-01

    In vivo homologous recombination holds the potential for optimal molecular cloning, however, current strategies require specialised bacterial strains or laborious protocols. Here, we exploit a recA-independent recombination pathway, present in widespread laboratory E.coli strains, to develop IVA (In Vivo Assembly) cloning. This system eliminates the need for enzymatic assembly and reduces all molecular cloning procedures to a single-tube, single-step PCR, performed in <2 hours from setup to transformation. Unlike other methods, IVA is a complete system, and offers significant advantages over alternative methods for all cloning procedures (insertions, deletions, site-directed mutagenesis and sub-cloning). Significantly, IVA allows unprecedented simplification of complex cloning procedures: five simultaneous modifications of any kind, multi-fragment assembly and library construction are performed in approximately half the time of current protocols, still in a single-step fashion. This system is efficient, seamless and sequence-independent, and requires no special kits, enzymes or proprietary bacteria, which will allow its immediate adoption by the academic and industrial molecular biology community. PMID:27264908

  5. IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly.

    PubMed

    García-Nafría, Javier; Watson, Jake F; Greger, Ingo H

    2016-01-01

    In vivo homologous recombination holds the potential for optimal molecular cloning, however, current strategies require specialised bacterial strains or laborious protocols. Here, we exploit a recA-independent recombination pathway, present in widespread laboratory E.coli strains, to develop IVA (In Vivo Assembly) cloning. This system eliminates the need for enzymatic assembly and reduces all molecular cloning procedures to a single-tube, single-step PCR, performed in <2 hours from setup to transformation. Unlike other methods, IVA is a complete system, and offers significant advantages over alternative methods for all cloning procedures (insertions, deletions, site-directed mutagenesis and sub-cloning). Significantly, IVA allows unprecedented simplification of complex cloning procedures: five simultaneous modifications of any kind, multi-fragment assembly and library construction are performed in approximately half the time of current protocols, still in a single-step fashion. This system is efficient, seamless and sequence-independent, and requires no special kits, enzymes or proprietary bacteria, which will allow its immediate adoption by the academic and industrial molecular biology community. PMID:27264908

  6. Innovation @ NASA

    NASA Technical Reports Server (NTRS)

    Roman, Juan A.

    2014-01-01

    This presentation provides an overview of the activities National Aeronautics and Space Administration (NASA) is doing to encourage innovation across the agency. All information provided is available publicly.

  7. Reply to the comments on "Comparison of Mann-Kendall and innovative trend method for water quality parameters of the Kizilirmak River, Turkey" by Kisi, O. and Ay, M. [J. Hydrol. 513 (2014) 362-375] and "An innovative method for trend analysis of monthly pan evaporations" by Kisi, O. [J. Hydrol. 527 (2015) 1123-1129

    NASA Astrophysics Data System (ADS)

    Kisi, Ozgur; Ay, Murat

    2016-07-01

    We thank discusser for his comments on our recent papers about application of innovative trend method (Şen, 2012) for water quality parameters (Kisi and Ay, 2014) and evaporation (Kisi, 2015) and are grateful for having the opportunity to explain and discuss the concerns raised. Illustration of the innovative Şen's trend method with some trend possibilities in the comments by discusser (in Fig. 3 in his discussion) has simple data seem to have normal distribution. However, some data may have highly skewed distribution and indicating trends in these data similar to the Figure 3 as suggested by discusser is impossible. Innovative trend analysis (Şen, 2012) of annual low flows for the duration 14-day (Q14) and 60-day (Q60) in Peterek and Dutdere stations are illustrated in Fig. 1 as an example. In Fig. 1, both half time series are sorted in ascending order. Therefore, Kisi and Ay (2014) provided "a general view" or "imaginative illustration" for the innovative Şen's trend method in Fig. 2 in their study. By this figure, general trend possibilities for any data having different distributions can be shown.

  8. Molecular profiling of microbial communities from contaminated sources: Use of subtractive cloning methods and rDNA spacer sequences. 1998 annual progress report

    SciTech Connect

    Robb, F.T.

    1998-06-01

    'The major objective of the research is to provide appropriate sequences and to assemble a high-density DNA array of oligonucleotides that can be used for rapid profiling of microbial populations from polluted areas. The sequences to be assigned to the DNA array are chosen from from cloned genomic DNA sequences (the ribosomal operon, described below) from groundwater at DOE sites containing organic solvents. The sites, Hanford Nuclear Plant and Lawrence Livermore Site 300, have well characterized pollutant histories, which have been provided by the collaborators. At this mid-point of the project, over 60 unique sequence classes of intergenic spacer region have been idedntified from the first sample site. The use of these sequences as hybridization probes, and their frequency of occurrence, allow a clear distinction between bacterial communities before and after remediation by acetate/nitrate pumping. The authors have developed the hybridization conditions for identifying PCR products in a 96 well format, a versatile alignment and visualization program (acronym: MALIGN) developed by Dr. Dennis Maeder, has been used to align the ISRs, which are variable in length and sometimes in position of the tRNAs. Finally, in collaboration with Dr. W. Chen and Dr. J. Zhou at ORNL, they have significant evidence that mass spectrometer analysis can be used to determine the lengths of PCR amplified intergenic spacer DNA.'

  9. Cloning of Homo sapiens? No!

    PubMed

    McKinnell, Robert G

    2002-01-01

    Animal cloning by nuclear transplantation was first developed in the northern leopard frog, Rana pipiens. It was soon extended to other amphibian species and within time, to various mammalian species. The production of a cloned sheep (Dolly) from an adult nuclear donor reawakened interest in human cloning. Nuclear transfer for the production of animal clones has served experimental biology well. Nonetheless, the potential burden of developmental hazards, scientists and funds diverted from more needy causes, as well as the potential assault on the concept of family has led the author to oppose human cloning.

  10. Cloning of Homo sapiens? No!

    PubMed

    McKinnell, Robert G

    2002-01-01

    Animal cloning by nuclear transplantation was first developed in the northern leopard frog, Rana pipiens. It was soon extended to other amphibian species and within time, to various mammalian species. The production of a cloned sheep (Dolly) from an adult nuclear donor reawakened interest in human cloning. Nuclear transfer for the production of animal clones has served experimental biology well. Nonetheless, the potential burden of developmental hazards, scientists and funds diverted from more needy causes, as well as the potential assault on the concept of family has led the author to oppose human cloning. PMID:11841468

  11. [Media, cloning, and bioethics].

    PubMed

    Costa, S I; Diniz, D

    2000-01-01

    This article was based on an analysis of three hundred articles from mainstream Brazilian periodicals over a period of eighteen months, beginning with the announcement of the Dolly case in February 1997. There were two main objectives: to outline the moral constants in the press associated with the possibility of cloning human beings and to identify some of the moral assumptions concerning scientific research with non-human animals that were published carelessly by the media. The authors conclude that there was a haphazard spread of fear concerning the cloning of human beings rather than an ethical debate on the issue, and that there is a serious gap between bioethical reflections and the Brazilian media.

  12. Towards an understanding of British public attitudes concerning human cloning.

    PubMed

    Shepherd, Richard; Barnett, Julie; Cooper, Helen; Coyle, Adrian; Moran-Ellis, Jo; Senior, Victoria; Walton, Chris

    2007-07-01

    The ability of scientists to apply cloning technology to humans has provoked public discussion and media coverage. The present paper reports on a series of studies examining public attitudes to human cloning in the UK, bringing together a range of quantitative and qualitative methods to address this question. These included a nationally representative survey, an experimental vignette study, focus groups and analyses of media coverage. Overall the research presents a complex picture of attitude to and constructions of human cloning. In all of the analyses, therapeutic cloning was viewed more favourably than reproductive cloning. However, while participants in the focus groups were generally negative about both forms of cloning, and this was also reflected in the media analyses, quantitative results showed more positive responses. In the quantitative research, therapeutic cloning was generally accepted when the benefits of such procedures were clear, and although reproductive cloning was less accepted there was still substantial support. Participants in the focus groups only differentiated between therapeutic and reproductive cloning after the issue of therapeutic cloning was explicitly raised; initially they saw cloning as being reproductive cloning and saw no real benefits. Attitudes were shown to be associated with underlying values associated with scientific progress rather than with age, gender or education, and although there were a few differences in the quantitative data based on religious affiliation, these tended to be small effects. Likewise in the focus groups there was little direct appeal to religion, but the main themes were 'interfering with nature' and the 'status of the embryo', with the latter being used more effectively to try to close down further discussion. In general there was a close correspondence between the media analysis and focus group responses, possibly demonstrating the importance of media as a resource, or that the media reflect

  13. Factors influencing the commercialisation of cloning in the pork industry.

    PubMed

    Pratt, S L; Sherrer, E S; Reeves, D E; Stice, S L

    2006-01-01

    Production of cloned pigs using somatic cell nuclear transfer (SCNT) is a repeatable and predictable procedure and multiple labs around the world have generated cloned pigs and genetically modified cloned pigs. Due to the integrated nature of the pork production industry, pork producers are the most likely to benefit and are in the best position to introduce cloning in to production systems. Cloning can be used to amplify superior genetics or be used in conjunction with genetic modifications to produce animals with superior economic traits. Though unproven, cloning could add value by reducing pig-to-pig variability in economically significant traits such as growth rate, feed efficiency, and carcass characteristics. However, cloning efficiencies using SCNT are low, but predictable. The inefficiencies are due to the intrusive nature of the procedure, the quality of oocytes and/or the somatic cells used in the procedure, the quality of the nuclear transfer embryos transferred into recipients, pregnancy rates of the recipients, and neonatal survival of the clones. Furthermore, in commercial animal agriculture, clones produced must be able to grow and thrive under normal management conditions, which include attainment of puberty and subsequent capability to reproduce. To integrate SCNT into the pork industry, inefficiencies at each step of the procedure must be overcome. In addition, it is likely that non-surgical embryo transfer will be required to deliver cloned embryos, and/or additional methods to generate high health clones will need to be developed. This review will focus on the state-of-the-art for SCNT in pigs and the steps required for practical implementation of pig cloning in animal agriculture.

  14. Probabilistic cloning of equidistant states

    SciTech Connect

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

    2010-08-15

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

  15. Ethical issues in livestock cloning.

    PubMed

    Thompson, P B

    1999-01-01

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

  16. Ethical issues in livestock cloning.

    PubMed

    Thompson, P B

    1999-01-01

    Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity. PMID:15719505

  17. Bounds for state-dependent quantum cloning

    SciTech Connect

    Han Yongjian; Zhang Yongsheng; Guo Guangcan

    2002-11-01

    Due to the no-cloning theorem, the unknown quantum state can only be cloned approximately or exactly with some probability. There are two types of cloners: universal and state-dependent cloner. The optimal universal cloner has been found and can be viewed as a special state-dependent quantum cloner that has no information about the states. In this paper, we investigate the state-dependent cloning when the state set contains more than two states. We get some bounds of the global fidelity for these processes. This method is not dependent on the number of the states contained in the state set. It is also independent of the numbers of copying.

  18. Cloning-free CRISPR

    PubMed Central

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

    2015-01-01

    Summary 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 target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate) at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate) through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis. PMID:26527385

  19. Cloning: Learning to Replay the Genetic Tape.

    ERIC Educational Resources Information Center

    Holden, David J.

    1979-01-01

    Describes how plants can be produced by cloning by using tissue culture methods to mass-produce rare native prairie plants and trying to transfer some of the genetic characteristics of native grasses into cultivated cereals. The experiment was conducted at South Dakota State University. (HM)

  20. Robert Koch: the grandfather of cloning?

    PubMed

    Weiss, Robin A

    2005-11-18

    This year marks the centenary of Robert Koch's Nobel Prize for discovering the cause of tuberculosis. Koch was also the first scientist to isolate the anthrax and cholera microbes. Yet perhaps one of his greatest contributions to biology is the least appreciated: his method for propagating individual colonies of bacteria on plates, a technique that came to be called cloning.

  1. Managing Innovation.

    ERIC Educational Resources Information Center

    Mason, Marilyn Gell

    1991-01-01

    Describes one library's use of technological innovation to provide Associated Press updates over the library's 24-hour dial-up service during the Persian Gulf War. Suggests five rules for innovation: (1) develop a shared vision; (2) foster frequent, formal, and informal communication; (3) empower employees; (4) take limited risks; and (5) use, but…

  2. Innovative Clinical Trial Designs

    PubMed Central

    Lavori, Philip W.

    2015-01-01

    Whereas the 20th-century health care system sometimes seemed to be inhospitable to and unmoved by experimental research, its inefficiency and unaffordability have led to reforms that foreshadow a new health care system. We point out certain opportunities and transformational needs for innovations in study design offered by the 21st-century health care system, and describe some innovative clinical trial designs and novel design methods to address these needs and challenges. PMID:26140056

  3. Use of an Innovative Simple Method for Anaerobiosis in the Diagnosis and Management of Infections in Two Unusual Cases.

    PubMed

    Haldar, J; Pal, N; Ray, R; Maiti, P K

    2016-01-01

    Technical limitations restrict routine anaerobe isolation from clinical materials in resource-limited laboratories. An innovative two steps combustion candle jar technique may be suitable for such setup. This system was tried with one case of chronic osteomyelitis developed on supracondyler compound fracture. Porphyromonas spp. was isolated and identified. Vancomycin was recommended based on in vitro sensitivity test, but the leg was amputed after receiving a resistant drug gentamycin. While in another child with hydrocephalous, V-P shunt associated infection by Peptostreptococcus anaerobius was successfully controlled by sensitive drug vancomycin. These two eye-opener cases insisted us for large scale application of the technique. PMID:27144078

  4. Mean-field methods in evolutionary duplication-innovation-loss models for the genome-level repertoire of protein domains

    NASA Astrophysics Data System (ADS)

    Angelini, A.; Amato, A.; Bianconi, G.; Bassetti, B.; Cosentino Lagomarsino, M.

    2010-02-01

    We present a combined mean-field and simulation approach to different models describing the dynamics of classes formed by elements that can appear, disappear, or copy themselves. These models, related to a paradigm duplication-innovation model known as Chinese restaurant process, are devised to reproduce the scaling behavior observed in the genome-wide repertoire of protein domains of all known species. In view of these data, we discuss the qualitative and quantitative differences of the alternative model formulations, focusing in particular on the roles of element loss and of the specificity of empirical domain classes.

  5. Interactive Methods for Teaching Action Potentials, an Example of Teaching Innovation from Neuroscience Postdoctoral Fellows in the Fellowships in Research and Science Teaching (FIRST) Program

    PubMed Central

    Keen-Rhinehart, E.; Eisen, A.; Eaton, D.; McCormack, K.

    2009-01-01

    Acquiring a faculty position in academia is extremely competitive and now typically requires more than just solid research skills and knowledge of one’s field. Recruiting institutions currently desire new faculty that can teach effectively, but few postdoctoral positions provide any training in teaching methods. Fellowships in Research and Science Teaching (FIRST) is a successful postdoctoral training program funded by the National Institutes of Health (NIH) providing training in both research and teaching methodology. The FIRST program provides fellows with outstanding interdisciplinary biomedical research training in fields such as neuroscience. The postdoctoral research experience is integrated with a teaching program which includes a How to Teach course, instruction in classroom technology and course development and mentored teaching. During their mentored teaching experiences, fellows are encouraged to explore innovative teaching methodologies and to perform science teaching research to improve classroom learning. FIRST fellows teaching neuroscience to undergraduates have observed that many of these students have difficulty with the topic of neuroscience. Therefore, we investigated the effects of interactive teaching methods for this topic. We tested two interactive teaching methodologies to determine if they would improve learning and retention of this information when compared with standard lectures. The interactive methods for teaching action potentials increased understanding and retention. Therefore, FIRST provides excellent teaching training, partly by enhancing the ability of fellows to integrate innovative teaching methods into their instruction. This training in turn provides fellows that matriculate from this program more of the characteristics that hiring institutions desire in their new faculty. PMID:23493377

  6. Rapid Construction of Recombinant Plasmids by QuickStep-Cloning.

    PubMed

    Jajesniak, Pawel; Wong, Tuck Seng

    2017-01-01

    QuickStep-Cloning is a novel molecular cloning technique that builds upon the concepts of asymmetric PCR and megaprimer-based amplification of whole plasmid. It was designed specifically to address the major drawbacks of previously reported cloning methods. The fully optimized protocol allows for a seamless integration of a long DNA fragment into any position within a plasmid of choice, in a time-efficient and cost-effective manner, without the need of a tedious DNA gel purification, a restriction digestion, and an enzymatic ligation. QuickStep-Cloning can be completed in less than 6 h, significantly faster than most of the existing cloning methods, while retaining high efficiency. PMID:27671943

  7. Cloned mice derived from somatic cell nuclei.

    PubMed

    Hosaka, K; Ohi, S; Ando, A; Kobayashi, M; Sato, K

    2000-12-01

    In 1997, a cloned sheep "Dolly" was produced by nuclear transfer of somatic cell. The first birth of cloned mice derived from some somatic cells were succeeded in 1998. At present, it is shown that somatic cells, cumulus cells, fibroblasts and Sertoli cells can be used to the study of cloned animal as nuclear donor. In this study investigation was designed to compare with efficiency on the production of cloned embryos by using the microinjection and the electrofusion methods for nuclear transfer. Oocyte enucleation was performed with a micromanipulator. The oocyte was held by holding pipette, and was enucleated using a beveled pipette. Microinjection method: Cell's nucleus injection was carried out by piezo-micromanipulator. Cytochalasin B treated cumulus cell was aspirated into a injection pipette, and was broken its plasma membrane using the injection pipette. Then, the cumulus cell was injected into the enucleated ooplasm directly. Electrofusion method: The cell was aspirated into a beveled pipette, and then an aspirated cell was inserted into perivitelline space. Then, the pair of enucleated oocyte and cell was fused using electrical cell fusion apparatus. The reconstituted embryos were activated after nuclear transfer using St2+. Reconstituted embryos had been produced by the microinjection showed the embryonic development to over 8-cell stages. But, the rate of fragmentation of reconstituted embryos by the microinjection showed a little high rate in comparison with the electrofusion. When some reconstituted embryos by the microinjection were transplanted to pseudopregnant females' oviduct, 9 fetuses were observed at 14 days post coitum. PMID:11329940

  8. Ethical issues in animal cloning.

    PubMed

    Fiester, Autumn

    2005-01-01

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

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

    PubMed

    Lupton, M L

    1999-01-01

    The cloning of Dolly the lamb from adult cells by scientists at the Roslin Laboratories near Edinburgh in February 1997 has startled the world because it now opens the way to clone adult human beings. The reaction to Ian Wilmut's breakthrough has been instant and largely negative. Bills were rushed into both the US Senate and House of Representatives aimed at banning the cloning of human beings. Human cloning is premature at this stage, but there are many positive spin-offs of cloning in the field of genetic engineering, such as the production of human proteins such as blood clotting factors which aid in healing wounds. Progress by means of cloning can also be made into devising a cure for Parkinson's Disease amongst others. No lesser ethicist than John C. Fletcher of the University of Virginia foresees circumstances in which human cloning is acceptable e.g. to enable a couple to replace a dying child, to enable a couple, one of whom is infertile, to clone a child from either partner. Extensive regulation of cloning by the law is inevitable but, in doing so, the legislation should be careful not to outlaw research in this area which could be beneficial to mankind. PMID:10436743

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

    PubMed

    Lupton, M L

    1999-01-01

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

  11. Lessons learned from cloning dogs.

    PubMed

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

    2012-08-01

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

  12. Therapeutic cloning and reproductive liberty.

    PubMed

    Sparrow, Robert

    2009-04-01

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

  13. Positional Cloning by Linkage Disequilibrium

    PubMed Central

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

    2004-01-01

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

  14. Cloning to reproduce desired genotypes.

    PubMed

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

    2001-01-01

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

  15. Human cloning and child welfare.

    PubMed Central

    Burley, J; Harris, J

    1999-01-01

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

  16. Therapeutic cloning: The ethical limits

    SciTech Connect

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

    2005-09-01

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

  17. Experimental reversion of the optimal quantum cloning and flipping processes

    SciTech Connect

    Sciarrino, Fabio; Secondi, Veronica; De Martini, Francesco

    2006-04-15

    The quantum cloner machine maps an unknown arbitrary input qubit into two optimal clones and one optimal flipped qubit. By combining linear and nonlinear optical methods we experimentally implement a scheme that, after the cloning transformation, restores the original input qubit in one of the output channels, by using local measurements, classical communication, and feedforward. This nonlocal method demonstrates how the information on the input qubit can be restored after the cloning process. The realization of the reversion process is expected to find useful applications in the field of modern multipartite quantum cryptography.

  18. Efficient preparation of shuffled DNA libraries through recombination (Gateway) cloning.

    PubMed

    Lehtonen, Soili I; Taskinen, Barbara; Ojala, Elina; Kukkurainen, Sampo; Rahikainen, Rolle; Riihimäki, Tiina A; Laitinen, Olli H; Kulomaa, Markku S; Hytönen, Vesa P

    2015-01-01

    Efficient and robust subcloning is essential for the construction of high-diversity DNA libraries in the field of directed evolution. We have developed a more efficient method for the subcloning of DNA-shuffled libraries by employing recombination cloning (Gateway). The Gateway cloning procedure was performed directly after the gene reassembly reaction, without additional purification and amplification steps, thus simplifying the conventional DNA shuffling protocols. Recombination-based cloning, directly from the heterologous reassembly reaction, conserved the high quality of the library and reduced the time required for the library construction. The described method is generally compatible for the construction of DNA-shuffled gene libraries.

  19. GENERAL: Linear Optical Scheme for Implementing Optimal Real State Cloning

    NASA Astrophysics Data System (ADS)

    Wan, Hong-Bo; Ye, Liu

    2010-06-01

    We propose an experimental scheme for implementing the optimal 1 → 3 real state cloning via linear optical elements. This method relies on one polarized qubit and two location qubits and is feasible with current experimental technology.

  20. [Placental developmental defects in cloned mammalian animals].

    PubMed

    Ao, Zheng; Liu, Dewu; Cai, Gengyuan; Wu, Zhenfang; Li, Zicong

    2016-05-01

    The cloning technique, also called somatic cell nuclear transfer (SCNT), has been successfully established and gradually applied to various mammalian species. However, the developmental rate of SCNT mammalian embryos is very low, usually at 1% to 5%, which limits the application of SCNT. Placental developmental defects are considered as the main cause of SCNT embryo development inhibition. Almost all of SCNT-derived mammalian placentas exhibit various abnormalities, such as placental hyperplasia, vascular defects and umbilical cord malformation. Mechanistically, these abnormalities result from failure of establishment of correct epigenetic modification in the trophectoderm genome, which leads to erroneous expression of important genes for placenta development-related, particularly imprinted genes. Consequently, aberrant imprinted gene expression gives rise to placental morphologic abnormalities and functional defects, therefore decreases developmental competence of cloned embryos. Currently, although numerous methods that can improve the developmental ability of SCNT-derived embryos have been reported, most of them are unable to substantially enhance the success rate of SCNT due to failure to eliminate the placental development defects. In this review, we summarize placental abnormalities and imprinted gene expression in mammalian cloning, and propose directions for the future research aiming to improve the cloning efficiency. PMID:27232488

  1. [The discrete horror of cloning].

    PubMed

    Guibourg, Ricardo A

    2009-01-01

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

  2. CATO: The Clone Alignment Tool.

    PubMed

    Henstock, Peter V; LaPan, Peter

    2016-01-01

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

  3. Animal Cloning and Food Safety

    MedlinePlus

    ... from clones and their offspring out of the food chain until CVM could further evaluate the issue. back to top FDA Studies Cloning For more than five years, CVM ... evaluate the safety of food from these animals. The resulting report, called a ...

  4. CATO: The Clone Alignment Tool.

    PubMed

    Henstock, Peter V; LaPan, Peter

    2016-01-01

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

  5. CATO: The Clone Alignment Tool

    PubMed Central

    Henstock, Peter V.; LaPan, Peter

    2016-01-01

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

  6. [The discrete horror of cloning].

    PubMed

    Guibourg, Ricardo A

    2009-01-01

    The author raises the topic of cloning after the decision of the Argentine government, which concerned for the "dignity of the human person", passed a decree of need and urgency, No. 200/97 (Annex), prohibiting cloning experiments with human beings. Therefore, considering that the topic is so terribly urgent and necessary, the author feels it is timely to consider it. PMID:19860340

  7. Therapeutic cloning: promises and issues

    PubMed Central

    Kfoury, Charlotte

    2007-01-01

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

  8. Course Management Systems and Blended Learning: An Innovative Learning Approach

    ERIC Educational Resources Information Center

    Chou, Amy Y.; Chou, David C.

    2011-01-01

    This article utilizes Rogers' innovation-decision process model (2003) and Beckman and Berry's innovation process model (2007) to create an innovative learning map that illustrates three learning methods (i.e., face-to-face learning, online learning, and blended learning) in two types of innovation (i.e., incremental innovation and radical…

  9. Animal cloning: problems and prospects.

    PubMed

    Wells, D N

    2005-04-01

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

  10. Network cloning unfolds the effect of clustering on dynamical processes

    NASA Astrophysics Data System (ADS)

    Faqeeh, Ali; Melnik, Sergey; Gleeson, James P.

    2015-05-01

    We introduce network L -cloning, a technique for creating ensembles of random networks from any given real-world or artificial network. Each member of the ensemble is an L -cloned network constructed from L copies of the original network. The degree distribution of an L -cloned network and, more importantly, the degree-degree correlation between and beyond nearest neighbors are identical to those of the original network. The density of triangles in an L -cloned network, and hence its clustering coefficient, is reduced by a factor of L compared to those of the original network. Furthermore, the density of loops of any fixed length approaches zero for sufficiently large values of L . Other variants of L -cloning allow us to keep intact the short loops of certain lengths. As an application, we employ these network cloning methods to investigate the effect of short loops on dynamical processes running on networks and to inspect the accuracy of corresponding tree-based theories. We demonstrate that dynamics on L -cloned networks (with sufficiently large L ) are accurately described by the so-called adjacency tree-based theories, examples of which include the message passing technique, some pair approximation methods, and the belief propagation algorithm used respectively to study bond percolation, SI epidemics, and the Ising model.

  11. Human cloning: can it be made safe?

    PubMed

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

    2003-11-01

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

  12. Real-time polymerase chain reaction method for detection of toxigenic Clostridium difficile from stools and presumptive identification of NAP1 clone.

    PubMed

    Jayaratne, Padman A; Monkman, Lori; Broukhanski, George; Pillai, Dillan R; Lee, Christine

    2013-02-01

    This study describes the development of a cost-effective, multiplex real-time polymerase chain reaction (RTPCR) method for detection of toxigenic Clostridium difficile from stools and presumptive identification of the NAP-1 strain. The diagnostic value of the new method is for the detection of toxigenic C. difficile which has the following performance characteristics: 99.8% specificity, 95.1% sensitivity, 97.5% positive predictive value, and 99.5% negative predictive value. Examination of 24 specimens presumptively identified as NAP1 strain by RTPCR with Pulsed-field gel electrophoresis performed on C. difficile isolated from those specimens showed 100% agreement. This RTPCR showed equivalent test performance characteristics as the 2 commercially available assays which were evaluated. The estimated cost per test is CAD$9.50 and which is significantly less than the commercial assays. The average turnaround time from setup to detection is 3.5 h. The RTPCR method described here is a cost-effective and highly sensitive test which can be implemented in a clinical laboratory to assist clinicians in establishing the diagnosis of C. difficile infection and indirectly determine the presence of the hypervirulent epidemic binary toxin (BI)/NAP 1 strain for prompt infection control interventions. PMID:23182075

  13. Innovation Networks

    NASA Astrophysics Data System (ADS)

    Pyka, Andreas; Scharnhorst, Andrea

    The idea for this book started when we organized a topical workshop entitled "Innovation Networks - New Approaches in Modeling and Analyzing" (held in Augsburg, Germany in October 2005), under the auspices of Exystence, a network of excellence funded in the European Union's Fifth Framework Program. Unlike other conferences on innovation and networks, however, this workshop brought together scientists from economics, sociology, communication science, science and technology studies, and physics. With this book we aim to build further on a bridge connecting the bodies of knowledge on networks in economics, the social sciences and, more recently, statistical physics.

  14. Hydrometry's classical and Innovative methods and tools comparison for Stara river flows at Agios Germanos monitoring station in north-west Greece.

    NASA Astrophysics Data System (ADS)

    Filintas, Agathos, , Dr; Hatzigiannakis, Evagellos, , Dr; Arampatzis, George, , Dr; Ilias, Andreas; Panagopoulos, Andreas, , Dr; Hatzispiroglou, Ioannis

    2015-04-01

    The aim of the present study is a thorough comparison of hydrometry's conventional and innovative methods-tools for river flow monitoring. A case study was conducted in Stara river at Agios Germanos monitoring station (northwest Greece), in order to investigate possible deviations between conventional and innovative methods-tools on river flow velocity and discharge. For this study, two flowmeters were used, which manufac-tured in 2013 (OTT Messtechnik Gmbh, 2013), as follows: a) A conventional propeller flow velocity meter (OTT-Model C2) which is a me-chanical current flow meter with a certification of calibration BARGO, operated with a rod and a relocating device, along with a digital measuring device including an elec-tronic flow calculator, data logger and real time control display unit. The flowmeter has a measurement velocity range 0.025-4.000 m/s. b) An innovative electromagnetic flowmeter (OTT-Model MF pro) which it is con-sisted of a compact and light-weight sensor and a robust handheld unit. Both system components are designed to be attached to conventional wading rods. The electromag-netic flowmeter uses Faraday's Law of electromagnetic induction to measure the process flow. When an electrically conductive fluid flows along the meter, an electrode voltage is induced between a pair of electrodes placed at right angles to the direction of mag-netic field. The electrode voltage is directly proportional to the average fluid velocity. The electromagnetic flowmeter was operated with a rod and relocating device, along with a digital measuring device with various logging and graphical capabilities and vari-ous methods of velocity measurement (ISO/USGS standards). The flowmeter has a measurement velocity range 0.000-6.000 m/s. The river flow data were averaged over a pair measurement of 60+60 seconds and the measured river water flow velocity, depths and widths of the segments were used for the estimation of cross-section's mean flow velocity in each measured

  15. Using an innovative combination of quality-by-design and green analytical chemistry approaches for the development of a stability indicating UHPLC method in pharmaceutical products.

    PubMed

    Boussès, Christine; Ferey, Ludivine; Vedrines, Elodie; Gaudin, Karen

    2015-11-10

    An innovative combination of green chemistry and quality by design (QbD) approach is presented through the development of an UHPLC method for the analysis of the main degradation products of dextromethorphan hydrobromide. QbD strategy was integrated to the field of green analytical chemistry to improve method understanding while assuring quality and minimizing environmental impacts, and analyst exposure. This analytical method was thoroughly evaluated by applying risk assessment and multivariate analysis tools. After a scouting phase aimed at selecting a suitable stationary phase and an organic solvent in accordance with green chemistry principles, quality risk assessment tools were applied to determine the critical process parameters (CPPs). The effects of the CPPs on critical quality attributes (CQAs), i.e., resolutions, efficiencies, and solvent consumption were further evaluated by means of a screening design. A response surface methodology was then carried out to model CQAs as function of the selected CPPs and the optimal separation conditions were determined through a desirability analysis. Resulting contour plots enabled to establish the design space (DS) (method operable design region) where all CQAs fulfilled the requirements. An experimental validation of the DS proved that quality within the DS was guaranteed; therefore no more robustness study was required before the validation. Finally, this UHPLC method was validated using the concept of total error and was used to analyze a pharmaceutical drug product. PMID:26183807

  16. Using an innovative combination of quality-by-design and green analytical chemistry approaches for the development of a stability indicating UHPLC method in pharmaceutical products.

    PubMed

    Boussès, Christine; Ferey, Ludivine; Vedrines, Elodie; Gaudin, Karen

    2015-11-10

    An innovative combination of green chemistry and quality by design (QbD) approach is presented through the development of an UHPLC method for the analysis of the main degradation products of dextromethorphan hydrobromide. QbD strategy was integrated to the field of green analytical chemistry to improve method understanding while assuring quality and minimizing environmental impacts, and analyst exposure. This analytical method was thoroughly evaluated by applying risk assessment and multivariate analysis tools. After a scouting phase aimed at selecting a suitable stationary phase and an organic solvent in accordance with green chemistry principles, quality risk assessment tools were applied to determine the critical process parameters (CPPs). The effects of the CPPs on critical quality attributes (CQAs), i.e., resolutions, efficiencies, and solvent consumption were further evaluated by means of a screening design. A response surface methodology was then carried out to model CQAs as function of the selected CPPs and the optimal separation conditions were determined through a desirability analysis. Resulting contour plots enabled to establish the design space (DS) (method operable design region) where all CQAs fulfilled the requirements. An experimental validation of the DS proved that quality within the DS was guaranteed; therefore no more robustness study was required before the validation. Finally, this UHPLC method was validated using the concept of total error and was used to analyze a pharmaceutical drug product.

  17. [Eros, Thanatos and a cloned child].

    PubMed

    Szewczyk, K

    2001-01-01

    The paper discusses and confirms the opinion that modern Western European culture is characterised by a high level of fear of death, which shows all features of a thanatic crisis. This is a consequence of wearing-out of culture-made means used to alleviate the fear induced by human finity. In this situation, modern societies put more and more hope in supported procreation and cloning of Homo sapiens as methods of reducing thanatic fears. PMID:11684774

  18. [Eros, Thanatos and a cloned child].

    PubMed

    Szewczyk, K

    2001-01-01

    The paper discusses and confirms the opinion that modern Western European culture is characterised by a high level of fear of death, which shows all features of a thanatic crisis. This is a consequence of wearing-out of culture-made means used to alleviate the fear induced by human finity. In this situation, modern societies put more and more hope in supported procreation and cloning of Homo sapiens as methods of reducing thanatic fears.

  19. Expanding the Aperture of Psychological Assessment: Introduction to the Special Section on Innovative Clinical Assessment Technologies and Methods

    ERIC Educational Resources Information Center

    Trull, Timothy J.

    2007-01-01

    Contemporary psychological assessment is dominated by tried-and-true methods like clinical interviewing, self-report questionnaires, intellectual assessment, and behavioral observation. These approaches have served as the mainstays of psychological assessment for decades. To be sure, these methods have survived over the years because clinicians…

  20. Wildlife conservation and reproductive cloning.

    PubMed

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

    2004-03-01

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

  1. Mathematical Innovation

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Through Small Business Innovation Research funds from the Stennis Space Center, MathSoft, Inc., developed a system that can provide the building blocks for signal analysis and rapid prototyping. The product is the result of work to help NASA develop a complete understanding propulsion test data by using time frequency displays, automatic estimation and denoising, and data analysis plots for wavelet decomposition.

  2. Accelerating Innovation

    ERIC Educational Resources Information Center

    Education Week, 2012

    2012-01-01

    The word "innovation" seems to be in everyone's lexicon these days; it's even turning up as part of new education job titles in school districts and states. The ideas that undergird it are animating a growing movement that's spurring new policies, programs, and products that carry with them the potential to transform how students learn and how…

  3. Therapeutic cloning in the mouse.

    PubMed

    Mombaerts, Peter

    2003-09-30

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

  4. Cloning: revisiting an old debate.

    PubMed

    Verhey, Allen D

    1994-09-01

    The debate about cloning that took place 25 years ago, although directed toward a different sort of cloning, elucidates fundamental issues currently at stake in reproductive technologies and research. Paul Ramsey and Joseph Fletcher were participants in this early debate. The differences between Ramsey and Fletcher about the meaning and sufficiency of freedom, the understanding and weighing of good and evil, the connection between embodiment and personhood, the relationship of humans with nature, and the meaning of parenthood suggest both a broader agenda for the debate about cloning and a cautious move forward in the development of embryo-splitting.

  5. Large Clones on Cliff Faces: Expanding by Rhizomes through Crevices

    PubMed Central

    Liu, Feng-Hong; Yu, Fei-Hai; Liu, Wen-Sheng; Krüsi, Bertil O.; Cai, Xiao-Hu; Schneller, Jakob J.; Dong, Ming

    2007-01-01

    Background and Aims Large clones of rhizomatous plants are found in many habitats, but little is known about whether such clones also occur on cliff faces where environmental conditions are extremely harsh and heterogeneous. Methods Using molecular (intersimple sequence repeat, ISSR) markers, the genotypic composition of a cliff-face population of Oxyria sinensis in Sichuan, China, was investigated. Key Results The 98 O. sinensis ramets sampled belonged to 12 different genotypes (clones). The three most frequent clones were represented with 45, 22 and 12 ramets, respectively; the remaining nine were represented with only one to five ramets. The three largest clones spanned at least 2·7 m in the vertical direction and 4·6–6·9 m in the horizontal direction on the cliff face. Conclusions On the cliff face, large clones of O. sinensis are formed by rhizomes growing along the crevices. Expansion by rhizomes may help O. sinensis to exploit the patchy resources and support establishment and growth of new ramets. Moreover, rooted ramets connected by rhizomes may effectively reduce the susceptibility of O. sinensis to rock fall and erosion and thus greatly improve the chances for long-term survival. The multi-clone structure indicates that sexual reproduction is also important for the long-term persistence of O. sinensis populations on cliffs. PMID:17525100

  6. Tradition and Innovation.

    ERIC Educational Resources Information Center

    Katter, Eldon, Ed.

    1995-01-01

    "The articles in this issue were selected because, in one way or another, they all touched on the notion of tradition and innovation." Storytelling and tribal dances are examples of past, traditional methods of passing cultural knowledge from elders to youth. Contemporary youth have replaced tradtional rites of passage with their own inventions…

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

    PubMed

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

    2006-01-01

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

  8. The Development and Use of an Innovative Laboratory Method for Measuring Arsenic in Drinking Water from Western Bangladesh

    PubMed Central

    Frisbie, Seth H.; Mitchell, Erika J.; Yusuf, Ahmad Zaki; Siddiq, Mohammad Yusuf; Sanchez, Raul E.; Ortega, Richard; Maynard, Donald M.; Sarkar, Bibudhendra

    2005-01-01

    All of Bangladesh’s approximately 10 million drinking-water tube wells must be periodically tested for arsenic. The magnitude of this task and the limited resources of Bangladesh have led to the use of low-cost, semiquantitative field kits that measure As to a relatively high 50 μg/L national drinking water standard. However, there is an urgent need to supplement and ultimately replace these field kits with an inexpensive laboratory method that can measure As to the more protective 10 μg/L World Health Organization (WHO) health-based drinking water guideline. Unfortunately, Bangladesh has limited access to atomic absorption spectrometers or other expensive instruments that can measure As to the WHO guideline of 10 μg/L. In response to this need, an inexpensive and highly sensitive laboratory method for measuring As has been developed. This new method is the only accurate, precise, and safe way to quantify As < 10 μg/L without expensive or highly specialized laboratory equipment. In this method, As is removed from the sample by reduction to arsine gas, collected in an absorber by oxidation to arsenic acid, colorized by a sequential reaction to arsenomolybdate, and quantified by spectrophotometry. We compared this method with the silver diethyldithiocarbamate [AgSCSN(CH2CH3)2] and graphite furnace atomic absorption spectroscopy (GFAAS) methods for measuring As. Our method is more accurate, precise, and environmentally safe than the AgSCSN(CH2CH3)2 method, and it is more accurate and affordable than GFAAS. Finally, this study suggests that Bangladeshis will readily share drinking water with their neighbors to meet the more protective WHO guideline for As of 10 μg/L. PMID:16140627

  9. An innovative method to measure the peripheral arterial elasticity: spring constant modeling based on the arterial pressure wave with radial vibration.

    PubMed

    Wei, Ching-Chuan

    2011-11-01

    In this study, we propose an innovative method for the direct measurement of the peripheral artery elasticity using a spring constant model, based on the arterial pressure wave equation, vibrating in a radial direction. By means of the boundary condition of the pressure wave equation at the maximum peak, we can derive the spring constant used for evaluating peripheral arterial elasticity. The calculated spring constants of six typical subjects show a coincidence with their proper arterial elasticities. Furthermore, the comparison between the spring constant method and pulse wave velocity (PWV) was investigated in 70 subjects (21-64 years, 47 normotensives and 23 hypertensives). The results reveal a significant negative correlation for the spring constant vs. PWV (correlation coefficient = -0.663, p < 0.001). Multivariate analysis also indicates the same close relationship. Furthermore, within-operator and between-operator analyses show significantly high reproducibility. Therefore, the use of the spring constant method to assess the arterial elasticity is carefully verified, and it is shown to be effective as well as fast. This method should be useful for healthcare, not only in improving clinical diagnosis of arterial stiffness but also in screening subjects for early evidence of cardio-vascular diseases and in monitoring responses to therapy in the future.

  10. Molecular profiling of microbial communities from contaminated sources: Use of substractive cloning methods and rDNA spacer sequences. 1997 annual progress report

    SciTech Connect

    1997-12-01

    'This project is to develop molecular methods for rapid characterization of microbial communities in contaminated ecosystems. The authors are exploring the use of {sup 16}s ribosomal DNA intergenic spacer regions (ISRs) to profile community composition. The choice proves to be a good one: there are 200--550 bases of 1 to 3 variable regions from which to choose species-specific probes, as well as 2--4 stretches of conserved sequence from which to develop universal PCR (polymerase chain reaction) primers. Preliminary community characterization is complete, and several types of arrays are under development to determine the types of bacteria present and the status of the ground water. Profiling the community composition of polluted groundwater will impact the broad field of microbial ecology as well as mixed-waste bioremediation. Results The samples the authors have been analysing were provided by Dr. Fred Brockman from Pacific Northwest Laboratory, and were collected at the US DOE Hanford site, Washington state. The samples were microbial filtrates from ground water polluted with 2 mg/L carbon tetrachloride and 250 mg/L nitrate and subjected to enrichment (acetate + nitrate) and recirculation. This project is described in some detail in PNNL-11113, Accelerated In Situ Bioremediation of Groundwater, by M.J. Truex, B.S. Hooker, and D.B. Anderson, July 1996.'

  11. Cloning and characterization of a cDNA clone encoding calreticulin from Haemaphysalis qinghaiensis (Acari: Ixodidae).

    PubMed

    Gao, Jinliang; Luo, Jianxun; Fan, Ruiquan; Fingerle, Volker; Guan, Guiquan; Liu, Zhijie; Li, Youquan; Zhao, Haiping; Ma, Miling; Liu, Junlong; Liu, Aihong; Ren, Qiaoyun; Dang, Zhisheng; Sugimoto, Chihiro; Yin, Hong

    2008-03-01

    The application of anti-tick vaccine has been shown to be the most promising alternative strategy compared to the current use of acaricides that suffer from a number of serious limitations. The success of this method is dependent upon identification and cloning of potential tick vaccine antigens. Previously, we have cloned 21 positive clones (named from Hq02 to Hq22) by immunoscreening complimentary DNA (cDNA) libraries of Haemaphysalis qinghaiensis; however, some of those clones did not contain open reading frames (ORF). In this study, we amplified the entire sequence of Hq07 by using rapid amplification of the cDNA ends. Hq07 contains an ORF of 1,233 bp that encodes for 410 amino acid residues with a coding capacity of 47 kDa. Search of the cloned sequences against GenBank revealed that Hq07 is a calreticulin (CRT)-similar clone and designated HqCRT. Expression analysis by reverse transcription-polymerase chain reaction showed that this gene is ubiquitously expressed at different developmental stages and in different tissues of H. qinghaiensis. The gene was expressed as glutathione S-transferase-fused proteins in a prokaryotic system. Western blot analysis revealed that native HqCRT was secreted into their hosts by ticks during blood sucking. Vaccination of sheep with rHqCRT conferred protective immunity against ticks, resulting in 54.3% mortality in adult ticks, compared to the 38.7% death rate in the control group. These results demonstrated that rHqCRT might be a useful vaccine candidate antigen for biological control of H. qinghaiensis.

  12. Methylotroph cloning vehicle

    DOEpatents

    Hanson, Richard S.; Allen, Larry N.

    1989-04-25

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

  13. Evaluation of a pooled strategy for high-throughput sequencing of cosmid clones from metagenomic libraries.

    PubMed

    Lam, Kathy N; Hall, Michael W; Engel, Katja; Vey, Gregory; Cheng, Jiujun; Neufeld, Josh D; Charles, Trevor C

    2014-01-01

    High-throughput sequencing methods have been instrumental in the growing field of metagenomics, with technological improvements enabling greater throughput at decreased costs. Nonetheless, the economy of high-throughput sequencing cannot be fully leveraged in the subdiscipline of functional metagenomics. In this area of research, environmental DNA is typically cloned to generate large-insert libraries from which individual clones are isolated, based on specific activities of interest. Sequence data are required for complete characterization of such clones, but the sequencing of a large set of clones requires individual barcode-based sample preparation; this can become costly, as the cost of clone barcoding scales linearly with the number of clones processed, and thus sequencing a large number of metagenomic clones often remains cost-prohibitive. We investigated a hybrid Sanger/Illumina pooled sequencing strategy that omits barcoding altogether, and we evaluated this strategy by comparing the pooled sequencing results to reference sequence data obtained from traditional barcode-based sequencing of the same set of clones. Using identity and coverage metrics in our evaluation, we show that pooled sequencing can generate high-quality sequence data, without producing problematic chimeras. Though caveats of a pooled strategy exist and further optimization of the method is required to improve recovery of complete clone sequences and to avoid circumstances that generate unrecoverable clone sequences, our results demonstrate that pooled sequencing represents an effective and low-cost alternative for sequencing large sets of metagenomic clones. PMID:24911009

  14. More than just fibers: an aqueous method for the production of innovative recombinant spider silk protein materials.

    PubMed

    Jones, Justin A; Harris, Thomas I; Tucker, Chauncey L; Berg, Kyle R; Christy, Stacia Y; Day, Breton A; Gaztambide, Danielle A; Needham, Nate J C; Ruben, Ashley L; Oliveira, Paula F; Decker, Richard E; Lewis, Randolph V

    2015-04-13

    Spider silk is a striking and robust natural material that has an unrivaled combination of strength and elasticity. There are two major problems in creating materials from recombinant spider silk proteins (rSSps): expressing sufficient quantities of the large, highly repetitive proteins and solvating the naturally self-assembling proteins once produced. To address the second problem, we have developed a method to rapidly dissolve rSSps in water in lieu of traditional organic solvents and accomplish nearly 100% solvation and recovery of the protein. Our method involves generating pressure and temperature in a sealed vial by using short, repetitive bursts from a conventional microwave. The method is scalable and has been successful with all rSSps used to date. From these easily generated aqueous solutions of rSSps, a wide variety of materials have been produced. Production of fibers, films, hydrogels, lyogels, sponges, and adhesives and studies of their mechanical and structural properties are reported. To our knowledge, ours is the only method that is cost-effective and scalable for mass production. This solvation method allows a choice of the physical form of product to take advantage of spider silks' mechanical properties without using costly and problematic organic solvents. PMID:25789668

  15. Innovative real-time and non-destructive method of beam profile measurement under large beam current irradiation for BNCT

    NASA Astrophysics Data System (ADS)

    Takada, M.; Kamada, S.; Suda, M.; Fujii, R.; Nakamura, M.; Hoshi, M.; Sato, H.; Endo, S.; Hamano, T.; Arai, S.; Higashimata, A.

    2012-10-01

    We developed a real-time and non-destructive method of beam profile measurement on a target under large beam current irradiation, and without any complex radiation detectors or electrical circuits. We measured the beam profiles on a target by observing the target temperature using an infrared-radiation thermometer camera. The target temperatures were increased and decreased quickly by starting and stopping the beam irradiation within 1 s in response speed. Our method could trace beam movements rapidly. The beam size and position were calibrated by measuring O-ring heat on the target. Our method has the potential to measure beam profiles at beam current over 1 mA for proton and deuteron with the energy around 3 MeV and allows accelerator operators to adjust the beam location during beam irradiation experiments without decreasing the beam current.

  16. Characterizing seamless ligation cloning extract for synthetic biological applications.

    PubMed

    Messerschmidt, Katrin; Hochrein, Lena; Dehm, Daniel; Schulz, Karina; Mueller-Roeber, Bernd

    2016-09-15

    Synthetic biology aims at designing and engineering organisms. The engineering process typically requires the establishment of suitable DNA constructs generated through fusion of multiple protein coding and regulatory sequences. Conventional cloning techniques, including those involving restriction enzymes and ligases, are often of limited scope, in particular when many DNA fragments must be joined or scar-free fusions are mandatory. Overlap-based-cloning methods have the potential to overcome such limitations. One such method uses seamless ligation cloning extract (SLiCE) prepared from Escherichia coli cells for straightforward and efficient in vitro fusion of DNA fragments. Here, we systematically characterized extracts prepared from the unmodified E. coli strain DH10B for SLiCE-mediated cloning and determined DNA sequence-associated parameters that affect cloning efficiency. Our data revealed the virtual absence of length restrictions for vector backbone (up to 13.5 kbp) and insert (90 bp to 1.6 kbp). Furthermore, differences in GC content in homology regions are easily tolerated and the deletion of unwanted vector sequences concomitant with targeted fragment insertion is straightforward. Thus, SLiCE represents a highly versatile DNA fusion method suitable for cloning projects in virtually all molecular and synthetic biology projects. PMID:27311554

  17. Innovating Firms and Aggregate Innovation.

    ERIC Educational Resources Information Center

    Klette, Tor Jakob; Kortum, Samuel

    2004-01-01

    We develop a parsimonious model of innovation to confront firm-level evidence. It captures the dynamics of individual heterogeneous firms, describes the behavior of an industry with firm entry and exit, and delivers a general equilibrium model of technological change. While unifying the theoretical analysis of firms, industries, and the aggregate…

  18. A Clone of Your Own.

    ERIC Educational Resources Information Center

    Bilodeau, Kirsten

    1997-01-01

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

  19. Cloning of a quantum measurement

    SciTech Connect

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

    2011-10-15

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

  20. Human cloning and 'posthuman' society.

    PubMed

    Blackford, Russell

    2005-01-01

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

  1. Are cloned quantum states macroscopic?

    PubMed

    Fröwis, F; Dür, W

    2012-10-26

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

  2. Human cloning and 'posthuman' society.

    PubMed

    Blackford, Russell

    2005-01-01

    Since early 1997, when the creation of Dolly the sheep by somatic cell nuclear transfer was announced in Nature, numerous government reports, essays, articles and books have considered the ethical problems and policy issues surrounding human reproductive cloning. In this article, I consider what response a modern liberal society should give to the prospect of human cloning, if it became safe and practical. Some opponents of human cloning have argued that permitting it would place us on a slippery slope to a repugnant future society, comparable to that portrayed in Aldous Huxley's novel, Brave New World. I conclude that, leaving aside concerns about safety, none of the psychological or social considerations discussed in this article provides an adequate policy justification for invoking the state's coercive powers to prevent human cloning. PMID:16007753

  3. Handmade somatic cell cloning in cattle.

    PubMed

    Vajta, Gàbor; Lewis, Ian M; Tecirlioglu, R Tayfur

    2006-01-01

    Apart from the biological and ethical problems, technical difficulties also hamper the improvement and widespread application of somatic cell nuclear transfer (NT). Recently introduced zona-free procedures may offer a solution for the latter problem. The most radical approach of these techniques is the so-called handmade cloning (HMC). It does not require micromanipulators because the manipulations required for both enucleation and nucleus transfer are performed by hand. The HMC technique includes manual bisection of zona-free oocytes, selection of cytoplasts by staining, and the simultaneous fusion of the somatic cell with two cytoplasts to produce a cloned embryo. HMC is a rapid and efficient technique that suits large-scale NT programs. It requires less expertise and time than traditional NT methods and the cost of equipment is significantly less. Production efficiency is high and embryo quality, in terms of pregnancy rates and live births, is not compromised. Although HMC has been developed particularly for bovine NT, the technique is applicable to other species. The method may become a useful tool for both experimental and commercial somatic cell cloning because it allows for standardization of procedures and provides the possibility of automation.

  4. Mapping genomic library clones using oligonucleotide arrays

    SciTech Connect

    Sapolsky, R.J.; Lipshutz, R.J.

    1996-05-01

    We have developed a high-density DNA probe array and accompanying biochemical and informatic methods to order clones from genomic libraries. This approach involves a series of enzymatic steps for capturing a set of short dispersed sequence markers scattered throughout a high-molecular-weight DNA. By this process, all the ambiguous sequences lying adjacent to a given Type IIS restriction site are ligated between two DNA adaptors. These markers, once amplified and labeled by PCR, can be hybridized and detected on a high-density olligonucleotide array bearing probes complementary to all possible markers. The array is synthesized using light-directed combinatorial chemistry. For each clone in a genomic library, a characteristic set of sequence markers can be determined. On the basis of the similarity between the marker sets for each pair of clones, their relative overlap can be measured. The library can be sequentially ordered into a contig map using this overlap information. This new methodology does not require gel-based methods or prior sequence information and involves manipulations that should allow for easy adaptation to automated processing and data collection. 28 refs., 9 figs., 2 tabs.

  5. Transport Phenomena Projects: Natural Convection between Porous, Concentric Cylinders--A Method to Learn and to Innovate

    ERIC Educational Resources Information Center

    Saatadjian, Esteban; Lesage, Francois; Mota, Jose Paulo B.

    2013-01-01

    A project that involves the numerical simulation of transport phenomena is an excellent method to teach this subject to senior/graduate chemical engineering students. The subject presented here has been used in our senior/graduate course, it concerns the study of natural convection heat transfer between two concentric, horizontal, saturated porous…

  6. Headspace Theater: An Innovative Method for Experiential Learning of Psychiatric Symptomatology Using Modified Role-Playing and Improvisational Theater Techniques

    ERIC Educational Resources Information Center

    Ballon, Bruce C.; Silver, Ivan; Fidler, Donald

    2007-01-01

    Objective: Headspace Theater has been developed to allow small group learning of psychiatric conditions by creating role-play situations in which participants are placed in a scenario that simulates the experience of the condition. Method: The authors conducted a literature review of role-playing techniques, interactive teaching, and experiential…

  7. Local cloning of entangled qubits

    SciTech Connect

    Choudhary, Sujit K.; Kunkri, Samir; Rahaman, Ramij; Roy, Anirban

    2007-11-15

    We discuss the exact cloning of orthogonal but entangled qubits under local operations and classical communication. The amount of entanglement necessary in a blank copy is obtained for various cases. Surprisingly, this amount is more than 1 ebit for certain sets of two nonmaximal but equally entangled states of two qubits. To clone any three Bell states, at least log{sub 2} 3 ebit is necessary.

  8. Cloning: questions answered and unsolved.

    PubMed

    Latham, Keith E

    2004-02-01

    Cloning by the transfer of adult somatic cell nuclei to oocytes has produced viable offspring in a variety of mammalian species. The technology is still in its initial stages of development. Studies to date have answered several basic questions related to such issues as genome potency, life expectancy of clones, mitochondrial fates, and feasibility of inter-species nuclear transfer. They have also raised new questions related to the control of nuclear reprogramming and function. These questions are reviewed here.

  9. Genomic mapping by end-characterized random clones: A mathematical analysis

    SciTech Connect

    Port, E.; Sun, F.; Martin, D.

    1995-03-01

    Physical maps can be constructed by {open_quotes}fingerprinting{close_quotes} a large number of random clones and inferring overlap between clones when the fingerprints are sufficiently similar. E. Lander and M. Waterman gave a mathematical analysis of such mapping strategies. The analysis is useful for comparing various fingerprinting methods. Recently it has been proposed that ends of clones rather than the entire clone be fingerprinted or characterized. Such fingerprints, which include sequenced clone ends, require a mathematical analysis deeper than that of Lander-Waterman. This paper studies clone islands, which can include uncharacterized regions, and also the islands that are formed entirely from the ends of clones. 23 refs., 11 figs., 4 tabs.

  10. Amphibian and mammal somatic-cell cloning: different species, common results?

    PubMed

    Loi, Pasqualino; Fulka, Josef; Ptak, Grazyna

    2003-11-01

    Since the production of Dolly the sheep cloning methods for somatic cells have been thoroughly described and are becoming routine. However, the rate at which live clones are produced remains low in all mammalian species tested so far. Remarkably, irrespective of the cloning protocol or the donor-cell type, all clones display common abnormalities, particularly in the placenta. The process is also complicated by early mortality of somatic-cell clones and the founder mammalian clone, Dolly the sheep, died in February 2003 aged six years. Based on published data and on our own experience, our view is that mammalian somatic-cell cloning and the pioneer nuclear-transfer data from amphibians have much in common. We suggest that the only way to improve nuclear reprogramming is to modify the chromatin structure of somatic cells before nuclear transfer, to provide the oocyte with a chromosomal structure that is more compatible with the natural reprogramming machinery of the oocyte.

  11. Reproductive ability of a cloned male detector dog and behavioral traits of its offspring

    PubMed Central

    Lee, Ji Hyun; Kim, Geon A; Kim, Rak Seung; Lee, Jong Su; Oh, Hyun Ju; Kim, Min Jung; Hong, Do Kyo

    2016-01-01

    In 2007, seven detector dogs were produced by somatic cell nuclear transfer using one nuclear donor dog, then trained and certified as excellent detector dogs, similar to their donor. In 2011, we crossed a cloned male and normal female by natural breeding and produced ten offspring. In this study, we investigated the puppies' temperaments, which we later compared with those of the cloned parent male. The results show that the cloned male had normal reproductive abilities and produced healthy offspring. All puppies completed narcotic detector dog training with a success rate for selection of 60%. Although the litter of cloned males was small in this study, a cloned male dog bred by natural mating produced puppies that later successfully completed the training course for drug detection. In conclusion, cloning an elite dog with superior genetic factors and breeding of the cloned dog was found to be a useful method to efficiently procure detector dogs. PMID:26435541

  12. Reproductive ability of a cloned male detector dog and behavioral traits of its offspring.

    PubMed

    Lee, Ji Hyun; Kim, Geon A; Kim, Rak Seung; Lee, Jong Su; Oh, Hyun Ju; Kim, Min Jung; Hong, Do Kyo; Lee, Byeong Chun

    2016-09-30

    In 2007, seven detector dogs were produced by somatic cell nuclear transfer using one nuclear donor dog, then trained and certified as excellent detector dogs, similar to their donor. In 2011, we crossed a cloned male and normal female by natural breeding and produced ten offspring. In this study, we investigated the puppies' temperaments, which we later compared with those of the cloned parent male. The results show that the cloned male had normal reproductive abilities and produced healthy offspring. All puppies completed narcotic detector dog training with a success rate for selection of 60%. Although the litter of cloned males was small in this study, a cloned male dog bred by natural mating produced puppies that later successfully completed the training course for drug detection. In conclusion, cloning an elite dog with superior genetic factors and breeding of the cloned dog was found to be a useful method to efficiently procure detector dogs.

  13. Amphibian and mammal somatic-cell cloning: different species, common results?

    PubMed

    Loi, Pasqualino; Fulka, Josef; Ptak, Grazyna

    2003-11-01

    Since the production of Dolly the sheep cloning methods for somatic cells have been thoroughly described and are becoming routine. However, the rate at which live clones are produced remains low in all mammalian species tested so far. Remarkably, irrespective of the cloning protocol or the donor-cell type, all clones display common abnormalities, particularly in the placenta. The process is also complicated by early mortality of somatic-cell clones and the founder mammalian clone, Dolly the sheep, died in February 2003 aged six years. Based on published data and on our own experience, our view is that mammalian somatic-cell cloning and the pioneer nuclear-transfer data from amphibians have much in common. We suggest that the only way to improve nuclear reprogramming is to modify the chromatin structure of somatic cells before nuclear transfer, to provide the oocyte with a chromosomal structure that is more compatible with the natural reprogramming machinery of the oocyte. PMID:14573358

  14. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

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

  15. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

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

  16. Artificial cloning of domestic animals

    PubMed Central

    Keefer, Carol L.

    2015-01-01

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

  17. Cloning goes to the movies.

    PubMed

    Cormick, Craig

    2006-10-01

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

  18. Islamic perspectives on human cloning.

    PubMed

    Sadeghi, Mahmoud

    2007-01-01

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

  19. Cloning goes to the movies.

    PubMed

    Cormick, Craig

    2006-10-01

    Public attitude research conducted by Biotechnology Australia shows that one of the major sources of information on human reproductive cloning is movies. Traditionally, understanding of new and emerging technologies has come through the mass media but human cloning, being so widely addressed through the popular culture of movies, is more effectively defined by Hollywood than the news media or science media. But how well are the science and social issues of cloning portrayed in box office hits such as The Island, Multiplicity, Star Wars: Attack of the Clones and Jurassic Park? These movies have enormous reach and undoubted influence, and are therefore worth analyzing in some detail. This study looks at 33 movies made between 1971 and 2005 that address human reproductive cloning, and it categorizes the films based on their genre and potential influence. Yet rather than simply rating the quality of the science portrayed, the study compares the key messages in these movies with public attitudes towards cloning, to examine the correlations. PMID:17214211

  20. Methylotroph cloning vehicle

    DOEpatents

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

    1989-04-25

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

  1. An Innovative Method of Measuring Changes in Access to Healthful Foods in School Lunch Programs: Findings from a Pilot Evaluation

    PubMed Central

    Hawkes, Allison P.; Weinberg, Stacy L.; Janusz, Ruth; Demont-Heinrich, Christine; Vogt, Richard L.

    2016-01-01

    Introduction A large local health department in Colorado partnered with 15 school districts to develop an approach to evaluate changes in access to healthy foods in reimbursable school lunches and a la carte offerings. Materials and Methods School district nutrition managers were engaged at the start of this project. Health department dietitians developed criteria to classify food items as “Lower Fat and less added Sugar” (LFS) and “Higher Fat and more added Sugar” (HFS) based on the percentage of calories from fat and grams of added sugar. Lunch production sheets were obtained for two time periods, food items and the number of planned servings recorded. LFS and HFS planned servings were summed for each time period, and a LFS to HFS ratio calculated by dividing LFS planned servings by HFS planned servings. Additional analyses included calculating LFS: HFS ratios by school district, and for a la carte offerings. Results In 2009, the LFS: HFS ratio was 2.08, in 2011, 3.71 (P<0.0001). The method also detected changes in ratios at the school district level. For a la carte items, in 2009 the ratio of LFS: HFS was 0.53, and in 2011, 0.61 (not statistically significant). Conclusions This method detected an increase in the LFS: HFS ratio over time and demonstrated that the school districts improved access to healthful food/drink by changing the contents of reimbursable school lunches. The evaluation method discussed here can generate information that districts can use in helping sustain and expand their efforts to create healthier environments for children and adults. Although federal regulations now cover all food and beverages served during the school day, there are still opportunities to improve and measure changes in food served in other settings such as child care centers, youth correction facilities, or in schools not participating in the National School Lunch Program. PMID:26800523

  2. Imperfect Cloning Operations in Algebraic Quantum Theory

    NASA Astrophysics Data System (ADS)

    Kitajima, Yuichiro

    2015-01-01

    No-cloning theorem says that there is no unitary operation that makes perfect clones of non-orthogonal quantum states. The objective of the present paper is to examine whether an imperfect cloning operation exists or not in a C*-algebraic framework. We define a universal -imperfect cloning operation which tolerates a finite loss of fidelity in the cloned state, and show that an individual system's algebra of observables is abelian if and only if there is a universal -imperfect cloning operation in the case where the loss of fidelity is less than . Therefore in this case no universal -imperfect cloning operation is possible in algebraic quantum theory.

  3. DNA cloning: A personal view after 40 years

    PubMed Central

    Cohen, Stanley N.

    2013-01-01

    In November 1973, my colleagues A. C. Y. Chang, H. W. Boyer, R. B. Helling, and I reported in PNAS that individual genes can be cloned and isolated by enzymatically cleaving DNA molecules into fragments, linking the fragments to an autonomously replicating plasmid, and introducing the resulting recombinant DNA molecules into bacteria. A few months later, Chang and I reported that genes from unrelated bacterial species can be combined and propagated using the same approach and that interspecies recombinant DNA molecules can produce a biologically functional protein in a foreign host. Soon afterward, Boyer’s laboratory and mine published our collaborative discovery that even genes from animal cells can be cloned in bacteria. These three PNAS papers quickly led to the use of DNA cloning methods in multiple areas of the biological and chemical sciences. They also resulted in a highly public controversy about the potential hazards of laboratory manipulation of genetic material, a decision by Stanford University and the University of California to seek patents on the technology that Boyer and I had invented, and the application of DNA cloning methods for commercial purposes. In the 40 years that have passed since publication of our findings, use of DNA cloning has produced insights about the workings of genes and cells in health and disease and has altered the nature of the biotechnology and biopharmaceutical industries. Here, I provide a personal perspective of the events that led to, and followed, our report of DNA cloning. PMID:24043817

  4. DNA cloning: a personal view after 40 years.

    PubMed

    Cohen, Stanley N

    2013-09-24

    In November 1973, my colleagues A. C. Y. Chang, H. W. Boyer, R. B. Helling, and I reported in PNAS that individual genes can be cloned and isolated by enzymatically cleaving DNA molecules into fragments, linking the fragments to an autonomously replicating plasmid, and introducing the resulting recombinant DNA molecules into bacteria. A few months later, Chang and I reported that genes from unrelated bacterial species can be combined and propagated using the same approach and that interspecies recombinant DNA molecules can produce a biologically functional protein in a foreign host. Soon afterward, Boyer's laboratory and mine published our collaborative discovery that even genes from animal cells can be cloned in bacteria. These three PNAS papers quickly led to the use of DNA cloning methods in multiple areas of the biological and chemical sciences. They also resulted in a highly public controversy about the potential hazards of laboratory manipulation of genetic material, a decision by Stanford University and the University of California to seek patents on the technology that Boyer and I had invented, and the application of DNA cloning methods for commercial purposes. In the 40 years that have passed since publication of our findings, use of DNA cloning has produced insights about the workings of genes and cells in health and disease and has altered the nature of the biotechnology and biopharmaceutical industries. Here, I provide a personal perspective of the events that led to, and followed, our report of DNA cloning.

  5. Improvement of the embalming perfusion method: the innovation and the results by light and scanning electron microscopy.

    PubMed

    Goyri-O'Neill, João; Pais, Diogo; Freire de Andrade, Francisco; Ribeiro, Paulo; Belo, Ana; O'Neill, Assunção; Ramos, Samuel; Neves Marques, Cláudia

    2013-01-01

    Embalming is a chemical process that aims the preservation and sanitization of the human body indefinitely. The technique of embalming is an important tool in teaching and research in anatomy enabling the preservation of cadaveric material in good conditions (lessening any significant structural changes and maintaining the natural appearance). This article presents the results of embalmed cadavers in the course of arterial perfusion, through the use of a perfusion machine, particularly designed to this objective, and which allows the control of the embalming fluid injection process. The influence of this technique and the optimization of its parameters on the final quality of embalming were evaluated by sequential histological analysis of the cadaveric tissues using an original method of classification of samples collected from 17 deceased corpses of the Corpses Donation Office of the Department of Anatomy of Faculdade de Ciências Médicas from Universidade Nova de Lisboa, subject to the embalming technique developed in the Department. We concluded that, with this method, there is a decrease of the decomposition process at the time of embalming, which is effective at long term (over a year), requiring merely the maintenance of the body at low temperatures (4° C) and it is possible to observe that the tissue best preserved over time is muscle, showing a conservation considered optimal.

  6. Innovative Method for Separating Phosphorus and Iron from High-Phosphorus Oolitic Hematite by Iron Nugget Process

    NASA Astrophysics Data System (ADS)

    Han, Hongliang; Duan, Dongping; Wang, Xing; Chen, Siming

    2014-10-01

    This study puts forward a new method to separate phosphorus and iron from high-phosphorus oolitic hematite through iron nuggets process. Firstly, the physical, chemical, and microscopic characteristics of high-phosphorus oolitic hematite are investigated. Then, the reaction mechanisms of high-phosphorus hematite together with feasibility to separating phosphorus and iron by iron nugget process are discussed. Meanwhile, the experiments of high-phosphorus hematite used in rotary hearth furnace iron nugget processes are studied as well. The results indicate that the iron nugget process is a feasible and efficient method for iron and phosphorus separation of high-phosphorus oolitic hematite. The phosphorus content in iron nuggets is relatively low. Through the optimization of process parameters, the lowest of phosphorus in iron nuggets is 0.22 pct, the dephosphorization rate is above 86 pct, and the recovery of Fe is above 85 pct by the iron nugget process. This study aims to provide a theoretical and technical basis for economical and rational use of high-phosphorus oolitic hematite.

  7. An innovative method for quality control of conjugated Haemophilus influenzae vaccines: A short review of two-dimensional nanoparticle electrophoresis.

    PubMed

    Tietz, Dietmar

    2009-12-25

    This article provides an overview of a 2D agarose electrophoretic procedure for the characterization of semi-synthetic Haemophilus influenzae type b meningitis vaccines that were prepared for the immunization of small children. The analysis of such vaccines has been particularly challenging because the vaccine particles (i) are highly negatively charged, (ii) are as large as or even larger than intact viruses, and (iii) have a continuous (polydisperse) size distribution because of randomizing steps in the vaccine production (sonification and crosslinking). As a result of these characteristics, 1D electrophoresis of the vaccines produced smears without discernable peaks, but with a second dimension of separation a characteristic vaccine fingerprint was obtained. Whereas O'Farrell gels can accomplish a 2D separation according to size and charge for samples with protein-sized particles, nondenaturing 2D agarose electrophoresis achieves a similar result for much larger virus-sized particles. The separation principle, however, is different. Even though the 2D electrophoretic method was developed from 1983 to 1995, it remains a promising tool for vaccine quality control and for predicting vaccine effectiveness. Modern technology makes the analysis significantly more practical and affordable than it was more than 10 years ago, and the method is applicable to a variety of conjugated vaccines and complex mixtures of virus-sized particles.

  8. Local cloning of two product states

    SciTech Connect

    Ji Zhengfeng; Feng Yuan; Ying Mingsheng

    2005-09-15

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

  9. Email-Based Informed Consent: Innovative Method for Reaching Large Numbers of Subjects for Data Mining Research

    NASA Technical Reports Server (NTRS)

    Lee, Lesley R.; Mason, Sara S.; Babiak-Vazquez, Adriana; Ray, Stacie L.; Van Baalen, Mary

    2015-01-01

    Since the 2010 NASA authorization to make the Life Sciences Data Archive (LSDA) and Lifetime Surveillance of Astronaut Health (LSAH) data archives more accessible by the research and operational communities, demand for data has greatly increased. Correspondingly, both the number and scope of requests have increased, from 142 requests fulfilled in 2011 to 224 in 2014, and with some datasets comprising up to 1 million data points. To meet the demand, the LSAH and LSDA Repositories project was launched, which allows active and retired astronauts to authorize full, partial, or no access to their data for research without individual, study-specific informed consent. A one-on-one personal informed consent briefing is required to fully communicate the implications of the several tiers of consent. Due to the need for personal contact to conduct Repositories consent meetings, the rate of consenting has not kept up with demand for individualized, possibly attributable data. As a result, other methods had to be implemented to allow the release of large datasets, such as release of only de-identified data. However the compilation of large, de-identified data sets places a significant resource burden on LSAH and LSDA and may result in diminished scientific usefulness of the dataset. As a result, LSAH and LSDA worked with the JSC Institutional Review Board Chair, Astronaut Office physicians, and NASA Office of General Counsel personnel to develop a "Remote Consenting" process for retrospective data mining studies. This is particularly useful since the majority of the astronaut cohort is retired from the agency and living outside the Houston area. Originally planned as a method to send informed consent briefing slides and consent forms only by mail, Remote Consenting has evolved into a means to accept crewmember decisions on individual studies via their method of choice: email or paper copy by mail. To date, 100 emails have been sent to request participation in eight HRP

  10. Predictability of some innovative behaviour in agricultural innovativeness.

    PubMed

    Das, P; Chakraborti, S; Gupta, T K

    1973-07-01

    An attempt is made to determine the relationship between a farmer's innovative behavior in the field of health, home, and family planning and his adoption of agricultural innovations. The study was carried out among all married farmers of the Muslim village of Padmerat in South Bengal territory. To measure the adoption behavior of the 196 farmers, 70 agricultural innovations were taken into account, 17 health practices, 10 family planning methods, and 22 home innovations. It was found that innovations in all 3 areas contributed toward agricultural innovative behavior, with family planning being the highest contributory, followed by home and health innovativeness. Together, these three can explain 62.15% of the total variation due to agricutural innovativeness. Income can explain another 16.91% of the variation, with the remaining part begin explained by other socioeconomic factors. It is recommended that agencies initiating programs to introduce technological change should work together to develop an integrated plan. This would bring better results at less cost.

  11. An Innovative Model: Undergraduate Poster Sessions by Health Professional Majors as a Method for Communicating Chemistry in Context

    NASA Astrophysics Data System (ADS)

    Dunstan, Marie; Bassinger, Pat

    1997-09-01

    This paper develops the methods used to conduct poster sessions for undergraduate health professional majors. There are two main goals of the project. The first goal is to learn chemistry in the context of health professions, everyday activities and contemporary issues. The second goal is to continue to develop research and written and oral communication skills. The topics are chosen after a classroom discussion of organic chemistry. The students then attend a library instruction class for introduction to appropriate research tools. Students are also taught the mechanics of poster preparation. The sessions are presented during a laboratory class period and are open to the college and public at large. Evaluation criteria include comprehension of the chemistry, the level and accuracy of the discussion and the appropriateness of the poster design for information exchange, All levels of the college community are invited to participate and greatly contribute to the success of the poster sessions.

  12. An Innovative Rapid Method for Analysis of 10 Organophosphorus Pesticide Residues in Wheat by HS-SPME-GC-FPD/MSD.

    PubMed

    Du, Xin; Ren, YongLin; Beckett, Stephen J

    2016-01-01

    The rapid detection of pesticide residues in wheat has become a top food security priority. A solvent-free headspace solid-phase microextraction (HS-SPME) has been evaluated for rapid screening of organophosphorus pesticide (OPP) residues in wheat with high sensitivity. Individual wheat samples (1.7 g), spiked with 10 OPPs, were placed in a 4 mL sealed amber glass vial and heated at 60°C for 45 min. During this time, the OPP residues were extracted with a 50 μm/30 μm divinylbenzene (DVB)/carboxen (CAR)/plasma desorption mass spectroscopy polydimethylsiloxane (PDMS) fiber from the headspace above the sample. The fiber was then removed and injected into the GC injection port at 250°C for desorption of the extracted chemicals. The multiple residues were identified by a GC mass spectrometer detector (GC-MSD) and quantified with a GC flame photometric detector (GC-FPD). Seven spiked levels of 10 OPPs on wheat were analyzed. The GC responses for a 50 μm/30 μm DVB/CAR/PDMS fiber increased with increasing spiking levels, yielding significant (R(2) > 0.98) linear regressions. The lowest LODs of the multiple pesticide standards were evaluated under the conditions of the validation study in a range of levels from 0 (control) to 100 ng of pesticide residue per g of wheat that separated on a low-polar GC capillary column (Agilent DB-35UI). The results of the HS-SPME method were compared with the QuEChERS AOAC 2007.01 method and they showed several advantages over the latter. These included improved sensitivity, selectivity, and simplicity. PMID:26964527

  13. Local cloning of entangled states

    SciTech Connect

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

    2010-08-15

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

  14. An innovative epitaxial growth method for minimizing dislocations in thin-film quantum-dot optoelectronic and photovoltaic device applications

    NASA Astrophysics Data System (ADS)

    Gandhi, Jateen S.

    A new buffer layer method for epitaxial growth of lattice-mismatched semiconductor quantum-dots based p-i-n structures is presented. To our knowledge this is the first instance of a dislocation-reduction approach that has shown reduced dark current behavior in a quantum-dot device compared to its counterpart homojunction p-n device consisted of the barrier material. The present work compared a lattice misfit strain build-up behavior between an In0.15Ga0.85As (p) / InAs ( i) / In0.15Ga0.85As (n) (QD) device to an In0.15Ga0.85As (p) / In 0.15Ga0.85As (n) (HOM) device, as both were grown on an un-doped gallium arsenide (GaAs) (100) substrate. The intrinsic region of QD device incorporated 5 layers of 2.1 ML indium arsenide quantum dots that were fabricated using self-assembly via Stranski-Krastanov strain release mechanism. Atomic force microscopy measurements exhibited 35 +/- 3 nm sized pyramidal islands with a narrow distribution and a density of 2.5 x 1010 per cm2. A low temperature (6K) photoluminescence characterization of the QD sample revealed an activity at 1400 nm wavelength that was attributed to optical pumping of carriers, which experienced a 3-dimensional quantum confinement due to a potential well formed by In0.15Ga0.85As matrix, and their subsequent radiative recombination. Both of the QD and HOM samples were characterized using x-ray diffractometer (XRD) and a high resolution transmission electron microscopy (HRTEM) method. The XRD data recorded a signature of the biaxially strained pseudomorphic section of In0.15Ga0.85As buffer layer that absorbed a lattice misfit due to epitaxial growth on GaAs substrate. This signature consisted of a set of twin peaks at higher and lower 2theta degrees resembling elastically strained and plastically relaxed sections, respectively, of the buffer layer residing in the vicinity of buffer-GaAs interface. A comparison of those peaks between QD and HOM samples exhibited an increase in the volume of the plastically relaxed

  15. Characterization of chlorinated solvent contamination in limestone using innovative FLUTe® technologies in combination with other methods in a line of evidence approach

    NASA Astrophysics Data System (ADS)

    Broholm, Mette M.; Janniche, Gry S.; Mosthaf, Klaus; Fjordbøge, Annika S.; Binning, Philip J.; Christensen, Anders G.; Grosen, Bernt; Jørgensen, Torben H.; Keller, Carl; Wealthall, Gary; Kerrn-Jespersen, Henriette

    2016-06-01

    Characterization of dense non-aqueous phase liquid (DNAPL) source zones in limestone aquifers/bedrock is essential to develop accurate site-specific conceptual models and perform risk assessment. Here innovative field methods were combined to improve determination of source zone architecture, hydrogeology and contaminant distribution. The FACT™ is a new technology and it was applied and tested at a contaminated site with a limestone aquifer, together with a number of existing methods including wire-line coring with core subsampling, FLUTe® transmissivity profiling and multilevel water sampling. Laboratory sorption studies were combined with a model of contaminant uptake on the FACT™ for data interpretation. Limestone aquifers were found particularly difficult to sample with existing methods because of core loss, particularly from soft zones in contact with chert beds. Water FLUTe™ multilevel groundwater sampling (under two flow conditions) and FACT™ sampling and analysis combined with FLUTe® transmissivity profiling and modeling were used to provide a line of evidence for the presence of DNAPL, dissolved and sorbed phase contamination in the limestone fractures and matrix. The combined methods were able to provide detailed vertical profiles of DNAPL and contaminant distributions, water flows and fracture zones in the aquifer and are therefore a powerful tool for site investigation. For the limestone aquifer the results indicate horizontal spreading in the upper crushed zone, vertical migration through fractures in the bryozoan limestone down to about 16-18 m depth with some horizontal migrations along horizontal fractures within the limestone. Documentation of the DNAPL source in the limestone aquifer was significantly improved by the use of FACT™ and Water FLUTe™ data.

  16. Characterization of chlorinated solvent contamination in limestone using innovative FLUTe® technologies in combination with other methods in a line of evidence approach.

    PubMed

    Broholm, Mette M; Janniche, Gry S; Mosthaf, Klaus; Fjordbøge, Annika S; Binning, Philip J; Christensen, Anders G; Grosen, Bernt; Jørgensen, Torben H; Keller, Carl; Wealthall, Gary; Kerrn-Jespersen, Henriette

    2016-06-01

    Characterization of dense non-aqueous phase liquid (DNAPL) source zones in limestone aquifers/bedrock is essential to develop accurate site-specific conceptual models and perform risk assessment. Here innovative field methods were combined to improve determination of source zone architecture, hydrogeology and contaminant distribution. The FACT™ is a new technology and it was applied and tested at a contaminated site with a limestone aquifer, together with a number of existing methods including wire-line coring with core subsampling, FLUTe® transmissivity profiling and multilevel water sampling. Laboratory sorption studies were combined with a model of contaminant uptake on the FACT™ for data interpretation. Limestone aquifers were found particularly difficult to sample with existing methods because of core loss, particularly from soft zones in contact with chert beds. Water FLUTe™ multilevel groundwater sampling (under two flow conditions) and FACT™ sampling and analysis combined with FLUTe® transmissivity profiling and modeling were used to provide a line of evidence for the presence of DNAPL, dissolved and sorbed phase contamination in the limestone fractures and matrix. The combined methods were able to provide detailed vertical profiles of DNAPL and contaminant distributions, water flows and fracture zones in the aquifer and are therefore a powerful tool for site investigation. For the limestone aquifer the results indicate horizontal spreading in the upper crushed zone, vertical migration through fractures in the bryozoan limestone down to about 16-18m depth with some horizontal migrations along horizontal fractures within the limestone. Documentation of the DNAPL source in the limestone aquifer was significantly improved by the use of FACT™ and Water FLUTe™ data. PMID:27116640

  17. Characterization of chlorinated solvent contamination in limestone using innovative FLUTe® technologies in combination with other methods in a line of evidence approach.

    PubMed

    Broholm, Mette M; Janniche, Gry S; Mosthaf, Klaus; Fjordbøge, Annika S; Binning, Philip J; Christensen, Anders G; Grosen, Bernt; Jørgensen, Torben H; Keller, Carl; Wealthall, Gary; Kerrn-Jespersen, Henriette

    2016-06-01

    Characterization of dense non-aqueous phase liquid (DNAPL) source zones in limestone aquifers/bedrock is essential to develop accurate site-specific conceptual models and perform risk assessment. Here innovative field methods were combined to improve determination of source zone architecture, hydrogeology and contaminant distribution. The FACT™ is a new technology and it was applied and tested at a contaminated site with a limestone aquifer, together with a number of existing methods including wire-line coring with core subsampling, FLUTe® transmissivity profiling and multilevel water sampling. Laboratory sorption studies were combined with a model of contaminant uptake on the FACT™ for data interpretation. Limestone aquifers were found particularly difficult to sample with existing methods because of core loss, particularly from soft zones in contact with chert beds. Water FLUTe™ multilevel groundwater sampling (under two flow conditions) and FACT™ sampling and analysis combined with FLUTe® transmissivity profiling and modeling were used to provide a line of evidence for the presence of DNAPL, dissolved and sorbed phase contamination in the limestone fractures and matrix. The combined methods were able to provide detailed vertical profiles of DNAPL and contaminant distributions, water flows and fracture zones in the aquifer and are therefore a powerful tool for site investigation. For the limestone aquifer the results indicate horizontal spreading in the upper crushed zone, vertical migration through fractures in the bryozoan limestone down to about 16-18m depth with some horizontal migrations along horizontal fractures within the limestone. Documentation of the DNAPL source in the limestone aquifer was significantly improved by the use of FACT™ and Water FLUTe™ data.

  18. Transforming beliefs and practices: Elementary teacher candidates' development through shared authentic teaching and reflection experiences within an innovative science methods course

    NASA Astrophysics Data System (ADS)

    Naidoo, Kara

    Elementary teachers are criticized for failing to incorporate meaningful science instruction in their classrooms or avoiding science instruction altogether. The lack of adequate science instruction in elementary schools is partially attributed to teacher candidates' anxiety, poor content and pedagogical preparation, and low science teaching self-efficacy. The central premise of this study was that many of these issues could be alleviated through course modifications designed to address these issues. The design tested and presented here provided prospective elementary educators' authentic science teaching experiences with elementary students in a low-stakes environment with the collaboration of peers and science teacher educators. The process of comprehensive reflection was developed for and tested in this study. Comprehensive reflection is individual and collective, written and set in dialogic discourse, focused on past and future behavior, and utilizes video recordings from shared teaching experiences. To test the central premise, an innovative science methods course was designed, implemented and evaluated using a one-group mixed-method design. The focus of the analysis was on changes in self-efficacy, identity and teaching practices as a function of authentic science teaching experiences and comprehensive reflection. The quantitative tools for analysis were t-tests and repeated-measures ANOVA on the Science Teaching Efficacy Belief Instrument-B (STEBI-B) and weekly self-rating on confidence as a learner and a teacher of science, respectively. The tools used to analyze qualitative data included thematic analysis and interpretative phenomenological analysis. In addition, theoretically grounded tools were developed and used in a case study to determine the ways one prospective educator's science teaching identity was influenced by experiences in the course. The innovative course structure led the development of teacher candidates' science teaching identity

  19. Classification of methods in transcranial Electrical Stimulation (tES) and evolving strategy from historical approaches to contemporary innovations

    PubMed Central

    Guleyupoglu, Berkan; Schestatsky, Pedro; Edwards, Dylan; Fregni, Felipe; Bikson, Marom

    2013-01-01

    Transcranial Electrical Stimulation (tES) encompasses all methods of non-invasive current application to the brain used in research and clinical practice. We present the first comprehensive and technical review, explaining the evolution of tES in both terminology and dosage over the past 100 years of research to present day. Current transcranial Pulsed Current Stimulation (tPCS) approaches such as Cranial Electrotherapy Stimulation (CES) descended from Electrosleep (ES) through Cranial Electro-stimulation Therapy (CET), Transcerebral Electrotherapy (TCET), and NeuroElectric Therapy (NET) while others like Transcutaneous Cranial Electrical Stimulation (TCES) descended from Electroanesthesia (EA) through Limoge, and Interferential Stimulation. Prior to a contemporary resurgence in interest, variations of trans-cranial Direct Current Stimulation were explored intermittently, including Polarizing current, Galvanic Vestibular Stimulation (GVS), and Transcranial Micropolarization. The development of these approaches alongside Electroconvulsive Therapy (ECT) and pharmacological developments are considered. Both the roots and unique features of contemporary approaches such as transcranial Alternating Current Stimulation (tACS) and transcranial Random Noise Stimulation (tRNS) are discussed. Trends and incremental developments in electrode montage and waveform spanning decades are presented leading to the present day. Commercial devices, seminal conferences, and regulatory decisions are noted. We conclude with six rules on how increasing medical and technological sophistication may now be leveraged for broader success and adoption of tES. PMID:23954780

  20. Integrated effect of gamma radiation and biocontrol agent on quality parameters of apple fruit: An innovative commercial preservation method

    NASA Astrophysics Data System (ADS)

    Ahari Mostafavi, Hossein; Mahyar Mirmajlessi, Seyed; Fathollahi, Hadi; Shahbazi, Samira; Mohammad Mirjalili, Seyed

    2013-10-01

    Effects of gamma irradiation and biocontrol agent (Pseudomonas fluorescens) on the physico-chemical parameters (including moisture, total soluble solids, antioxidant activity, phenolic content and firmness) of cv. Golden Delicious apples were investigated for their ability to avoid the post-harvest blue mold caused by Penicillium expansum during cold storage. Freshly harvested apples were inoculated with P. expansum. Treated fruits were irradiated at doses of 0, 200, 400, 600 and 800 Gy and then inoculated with P. fluorescens suspension. Samples were evaluated at 3 month intervals. The results demonstrated a clear link between antioxidant activity and phenolic content, so that dose range of 200-400 Gy significantly increased phenolic content and antioxidant activity. Effect of P. fluorescens was similar to irradiation at 200 and 400 Gy that could prevent lesion diameter in pathogen-treated apples. As dose and storage time increased firmness decreased but, combination of P. fluorescens as well as irradiation (at 200-400 Gy) could decrease softening apple fruits during storage. In all parameters, P. fluorescens (as biocontrol agent) inhibited P. expansum similar to irradiation at 200-400 Gy. So, integrated treatment of irradiation and biocontrol agent explored the potential dual benefit of low doses (200 and 400 Gy) as a suitable method to sustain physico-chemical quality and conclusively reduce apple fruits losses during post-harvest preservation.

  1. Classification of methods in transcranial electrical stimulation (tES) and evolving strategy from historical approaches to contemporary innovations.

    PubMed

    Guleyupoglu, Berkan; Schestatsky, Pedro; Edwards, Dylan; Fregni, Felipe; Bikson, Marom

    2013-10-15

    Transcranial Electrical Stimulation (tES) encompasses all methods of non-invasive current application to the brain used in research and clinical practice. We present the first comprehensive and technical review, explaining the evolution of tES in both terminology and dosage over the past 100 years of research to present day. Current transcranial Pulsed Current Stimulation (tPCS) approaches such as Cranial Electrotherapy Stimulation (CES) descended from Electrosleep (ES) through Cranial Electro-stimulation Therapy (CET), Transcerebral Electrotherapy (TCET), and NeuroElectric Therapy (NET) while others like Transcutaneous Cranial Electrical Stimulation (TCES) descended from Electroanesthesia (EA) through Limoge, and Interferential Stimulation. Prior to a contemporary resurgence in interest, variations of transcranial Direct Current Stimulation were explored intermittently, including Polarizing current, Galvanic Vestibular Stimulation (GVS), and Transcranial Micropolarization. The development of these approaches alongside Electroconvulsive Therapy (ECT) and pharmacological developments are considered. Both the roots and unique features of contemporary approaches such as transcranial Alternating Current Stimulation (tACS) and transcranial Random Noise Stimulation (tRNS) are discussed. Trends and incremental developments in electrode montage and waveform spanning decades are presented leading to the present day. Commercial devices, seminal conferences, and regulatory decisions are noted. We conclude with six rules on how increasing medical and technological sophistication may now be leveraged for broader success and adoption of tES.

  2. Optimal quantum cloning based on the maximin principle by using a priori information

    NASA Astrophysics Data System (ADS)

    Kang, Peng; Dai, Hong-Yi; Wei, Jia-Hua; Zhang, Ming

    2016-10-01

    We propose an optimal 1 →2 quantum cloning method based on the maximin principle by making full use of a priori information of amplitude and phase about the general cloned qubit input set, which is a simply connected region enclosed by a "longitude-latitude grid" on the Bloch sphere. Theoretically, the fidelity of the optimal quantum cloning machine derived from this method is the largest in terms of the maximin principle compared with that of any other machine. The problem solving is an optimization process that involves six unknown complex variables, six vectors in an uncertain-dimensional complex vector space, and four equality constraints. Moreover, by restricting the structure of the quantum cloning machine, the optimization problem is simplified as a three-real-parameter suboptimization problem with only one equality constraint. We obtain the explicit formula for a suboptimal quantum cloning machine. Additionally, the fidelity of our suboptimal quantum cloning machine is higher than or at least equal to that of universal quantum cloning machines and phase-covariant quantum cloning machines. It is also underlined that the suboptimal cloning machine outperforms the "belt quantum cloning machine" for some cases.

  3. [Cloning and law in Hungary].

    PubMed

    Julesz, Máté

    2015-03-01

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

  4. Upper-Extremity Dual-Task Function: An Innovative Method to Assess Cognitive Impairment in Older Adults

    PubMed Central

    Toosizadeh, Nima; Najafi, Bijan; Reiman, Eric M.; Mager, Reine M.; Veldhuizen, Jaimeson K.; O’Connor, Kathy; Zamrini, Edward; Mohler, Jane

    2016-01-01

    Background: Difficulties in orchestrating simultaneous tasks (i.e., dual-tasking) have been associated with cognitive impairments in older adults. Gait tests have been commonly used as the motor task component for dual-task assessments; however, many older adults have mobility impairments or there is a lack of space in busy clinical settings. We assessed an upper-extremity function (UEF) test as an alternative motor task to study the dual-task motor performance in older adults. Methods: Older adults (≥65 years) were recruited, and cognitive ability was measured using the Montreal cognitive assessment (MoCA). Participants performed repetitive elbow flexion with their maximum pace, once single-task, and once while counting backward by one (dual-task). Single- and dual-task gait tests were also performed with normal speed. Three-dimensional kinematics was measured both from upper-extremity and lower-extremity using wearable sensors to determine UEF and gait parameters. Parameters were compared between the cognitively impaired and healthy groups using analysis of variance tests, while controlling for age, gender, and body mass index (BMI). Correlations between UEF and gait parameters for dual-task and dual-task cost were assessed using linear regression models. Results: Sixty-seven older adults were recruited (age = 83 ± 10 years). Based on MoCA, 10 (15%) were cognitively impaired. While no significant differences were observed in the single-task condition, within the dual-task condition, the cognitively impaired group showed significantly less arm flexion speed (62%, d = 1.51, p = 0.02) and range of motion (27%, d = 0.93, p = 0.04), and higher speed variability (88%, d = 1.82, p < 0.0001) compared to the cognitively intact group, when adjusted with age, gender, and BMI. Significant correlations were observed between UEF speed parameters and gait stride velocity for dual-task condition (r = 0.55, p < 0.0001) and dual-task cost (r = 0.28, p = 0.03). Conclusion: We

  5. How to improve the success rate of mouse cloning technology.

    PubMed

    Thuan, Nguyen Van; Kishigami, Satoshi; Wakayama, Teruhiko

    2010-02-01

    It has now been 13 years since the first cloned mammal Dolly the sheep was generated from somatic cells using nuclear transfer (SCNT). Since then, this technique has been considered an important tool not only for animal reproduction but also for regenerative medicine. However, the success rate is still very low and the mechanisms involved in genomic reprogramming are not yet clear. Moreover, the NT technique requires donated fresh oocyte, which raises ethical problems for production of human cloned embryo. For this reason, the use of induced pluripotent stem cells for genomic reprogramming and for regenerative medicine is currently a hot topic in this field. However, we believe that the NT approach remains the only valid way for the study of reproduction and basic biology. For example, only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification, and it can generate offspring from a single cell or even a frozen dead body. Thanks to much hard work by many groups, cloning success rates are increasing slightly year by year, and NT cloning is now becoming a more applicable method. This review describes how to improve the efficiency of cloning, the establishment of clone-derived embryonic stem cells and further applications.

  6. Generating West Nile Virus from an Infectious Clone.

    PubMed

    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.

  7. How to improve the success rate of mouse cloning technology.

    PubMed

    Thuan, Nguyen Van; Kishigami, Satoshi; Wakayama, Teruhiko

    2010-02-01

    It has now been 13 years since the first cloned mammal Dolly the sheep was generated from somatic cells using nuclear transfer (SCNT). Since then, this technique has been considered an important tool not only for animal reproduction but also for regenerative medicine. However, the success rate is still very low and the mechanisms involved in genomic reprogramming are not yet clear. Moreover, the NT technique requires donated fresh oocyte, which raises ethical problems for production of human cloned embryo. For this reason, the use of induced pluripotent stem cells for genomic reprogramming and for regenerative medicine is currently a hot topic in this field. However, we believe that the NT approach remains the only valid way for the study of reproduction and basic biology. For example, only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification, and it can generate offspring from a single cell or even a frozen dead body. Thanks to much hard work by many groups, cloning success rates are increasing slightly year by year, and NT cloning is now becoming a more applicable method. This review describes how to improve the efficiency of cloning, the establishment of clone-derived embryonic stem cells and further applications. PMID:20203432

  8. The topsy-turvy cloning law.

    PubMed

    Brassington, Iain; Oultram, Stuart

    2011-03-01

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

  9. Innovative method to avoid the reduction of silver ions to silver nanoparticles \\left( A{{g}^{+}}\\to Ag{}^\\circ \\right) in silver ion conducting based polymer electrolytes

    NASA Astrophysics Data System (ADS)

    Aziz, Shujahadeen B.; Abidin, Zul Hazrin Z.; Kadir, M. F. Z.

    2015-03-01

    In this research work an innovative method is used to prevent the silver ion reduction in solid polymer electrolytes. The x-ray diffraction (XRD) results reveal the disruption of the crystalline nature of chitosan (CS) and formation of silver nanoparticles upon addition of silver triflate (AgTf) salt. The UV-vis measurement confirms the existence of silver nanoparticles via the broad surface plasmon resonance (SPR) peak. Upon the addition of Al2O3 nanoparticles the SPR peak intensity is greatly reduced. The amorphous domain of the CS:silver triflate (CS:AgTf) system increases with the addition of Al2O3 nanoparticles up to 4 wt.%. Deconvolution of the XRD results reveals that a larger crystallite size is obtained for higher Al2O3 concentrations and the peaks due to silver nanoparticles almost disappear. Scanning electron microscope (SEM) analyses show that Al2O3 nanoparticles are well dispersed at low concentrations and the leakage of chains of silver nanoparticles to the membrane surface almost disappear. The XRD, UV-vis, SEM and energy-dispersive x-ray (EDX) results strongly support that the reduction of silver ions to silver nanoparticles (Ag+ → Ag°) in the CS:silver triflate system is significantly avoided upon the addition of an Al2O3 filler.

  10. Development of an innovative 9 GHz EPR surface detection method and its application to non-invasive human fingers and nails investigation.

    PubMed

    Nakagawa, Kouichi

    2015-01-01

    We developed an innovative surface-type cavity for 9 GHz electron paramagnetic resonance (EPR) and used it to non-invasively measure human fingers and nails. This surface-type cavity measures a sample on the top of the cavity instead of a sample inserted into the cavity. To verify the performance of this method, 5-10 μL of 0.1 mM 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL) aqueous solution in a 1.0-mm (i.d.) glass capillary was used. Although the detectable radical concentration of the surface-type cavity is lower than that of a commercial cavity, the surface-type cavity can measure biomedical samples. In addition, the cavity is capable of measuring human fingers and nails. The dynamics of a topically applied 1.0-mM TEMPOL solution with a commercial lotion (5:1 by weight) on a human finger and nail were investigated. The present EPR results suggest that TEMPOL in solution may not easily permeate into the finger and nail during the measurements. Therefore, 9 GHz surface-type detection exhibits the good potential to study paramagnetic species in bulky biomedical samples.

  11. Development of an innovative 9 GHz EPR surface detection method and its application to non-invasive human fingers and nails investigation.

    PubMed

    Nakagawa, Kouichi

    2015-01-01

    We developed an innovative surface-type cavity for 9 GHz electron paramagnetic resonance (EPR) and used it to non-invasively measure human fingers and nails. This surface-type cavity measures a sample on the top of the cavity instead of a sample inserted into the cavity. To verify the performance of this method, 5-10 μL of 0.1 mM 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL) aqueous solution in a 1.0-mm (i.d.) glass capillary was used. Although the detectable radical concentration of the surface-type cavity is lower than that of a commercial cavity, the surface-type cavity can measure biomedical samples. In addition, the cavity is capable of measuring human fingers and nails. The dynamics of a topically applied 1.0-mM TEMPOL solution with a commercial lotion (5:1 by weight) on a human finger and nail were investigated. The present EPR results suggest that TEMPOL in solution may not easily permeate into the finger and nail during the measurements. Therefore, 9 GHz surface-type detection exhibits the good potential to study paramagnetic species in bulky biomedical samples. PMID:26067935

  12. The discipline of innovation. 1985.

    PubMed

    Drucker, Peter F

    2002-08-01

    How much of innovation is inspiration, and how much is hard work? The answer lies somewhere in the middle, says management thinker Peter Drucker. In this HBR classic from 1985, he argues that innovation is real work that can and should be managed like any other corporate function. Success is more likely to result from the systematic pursuit of opportunities than from a flash of genius. Indeed, most innovative business ideas arise through the methodical analysis of seven areas of opportunity. Within a company or industry, opportunities can be found in unexpected occurrences, incongruities of various kinds, process needs, or changes in an industry or market. Outside a company, opportunities arise from demographic changes, changes in perception, or new knowledge. There is some overlap among the sources, and the potential for innovation may well lie in more than one area at a time. Innovations based on new knowledge tend to have the greatest effect on the marketplace, but it often takes decades before the ideas are translated into actual products, processes, or services. The other sources of innovation are easier and simpler to handle, yet they still require managers to look beyond established practices, Drucker explains. The author emphasizes that innovators need to look for simple, focused solutions to real problems. The greatest praise an innovation can receive is for people to say, "That's so obvious!" Grandiose ideas designed to revolutionize an industry rarely work. Innovation, like any other endeavor, takes talent, ingenuity, and knowledge. But Drucker cautions that if diligence, persistence, and commitment are lacking, companies are unlikely to succeed at the business of innovation. PMID:12195923

  13. The discipline of innovation. 1985.

    PubMed

    Drucker, Peter F

    2002-08-01

    How much of innovation is inspiration, and how much is hard work? The answer lies somewhere in the middle, says management thinker Peter Drucker. In this HBR classic from 1985, he argues that innovation is real work that can and should be managed like any other corporate function. Success is more likely to result from the systematic pursuit of opportunities than from a flash of genius. Indeed, most innovative business ideas arise through the methodical analysis of seven areas of opportunity. Within a company or industry, opportunities can be found in unexpected occurrences, incongruities of various kinds, process needs, or changes in an industry or market. Outside a company, opportunities arise from demographic changes, changes in perception, or new knowledge. There is some overlap among the sources, and the potential for innovation may well lie in more than one area at a time. Innovations based on new knowledge tend to have the greatest effect on the marketplace, but it often takes decades before the ideas are translated into actual products, processes, or services. The other sources of innovation are easier and simpler to handle, yet they still require managers to look beyond established practices, Drucker explains. The author emphasizes that innovators need to look for simple, focused solutions to real problems. The greatest praise an innovation can receive is for people to say, "That's so obvious!" Grandiose ideas designed to revolutionize an industry rarely work. Innovation, like any other endeavor, takes talent, ingenuity, and knowledge. But Drucker cautions that if diligence, persistence, and commitment are lacking, companies are unlikely to succeed at the business of innovation.

  14. Genetic epidemiology, genetic maps and positional cloning.

    PubMed Central

    Morton, Newton E

    2003-01-01

    Genetic epidemiology developed in the middle of the last century, focused on inherited causes of disease but with methods and results applicable to other traits and even forensics. Early success with linkage led to the localization of genes contributing to disease, and ultimately to the Human Genome Project. The discovery of millions of DNA markers has encouraged more efficient positional cloning by linkage disequilibrium (LD), using LD maps and haplotypes in ways that are rapidly evolving. This has led to large international programmes, some promising and others alarming, with laws about DNA patenting and ethical guidelines for responsible research still struggling to be born. PMID:14561327

  15. In Defense of Innovative Methods.

    ERIC Educational Resources Information Center

    Parish, Charles

    A discussion of the growing assumption that the theories and approaches of the Silent Way, Counseling-Learning, Total Physical Response, Comprehension Training, and Suggestopedia deal with trivial issues rather than with the primary substance of language teaching and learning examines the question of rigorous scientific experimentation with…

  16. Quantum dot-based molecular imaging of cancer cell growth using a clone formation assay

    PubMed Central

    Geng, Xia-Fei; Fang, Min; Liu, Shao-Ping; Li, Yan

    2016-01-01

    This aim of the present study was to investigate clonal growth behavior and analyze the proliferation characteristics of cancer cells. The MCF-7 human breast cancer cell line, SW480 human colon cancer cell line and SGC7901 human gastric cancer cell line were selected to investigate the morphology of cell clones. Quantum dot-based molecular targeted imaging techniques (which stained pan-cytokeratin in the cytoplasm green and Ki67 in the cell nucleus yellow or red) were used to investigate the clone formation rate, cell morphology, discrete tendency, and Ki67 expression and distribution in clones. From the cell clone formation assay, the MCF-7, SW480 and SGC7901 cells were observed to form clones on days 6, 8 and 12 of cell culture, respectively. These three types of cells had heterogeneous morphology, large nuclear:cytoplasmic ratios, and conspicuous pathological mitotic features. The cells at the clone periphery formed multiple pseudopodium. In certain clones, cancer cells at the borderline were separated from the central cell clusters or presented a discrete tendency. With quantum dot-based molecular targeted imaging techniques, cells with strong Ki67 expression were predominantly shown to be distributed at the clone periphery, or concentrated on one side of the clones. In conclusion, cancer cell clones showed asymmetric growth behavior, and Ki67 was widely expressed in clones of these three cell lines, with strong expression around the clones, or aggregated at one side. Cell clone formation assay based on quantum dots molecular imaging offered a novel method to study the proliferative features of cancer cells, thus providing a further insight into tumor biology. PMID:27572664

  17. Quantum dot-based molecular imaging of cancer cell growth using a clone formation assay.

    PubMed

    Geng, Xia-Fei; Fang, Min; Liu, Shao-Ping; Li, Yan

    2016-10-01

    This aim of the present study was to investigate clonal growth behavior and analyze the proliferation characteristics of cancer cells. The MCF‑7 human breast cancer cell line, SW480 human colon cancer cell line and SGC7901 human gastric cancer cell line were selected to investigate the morphology of cell clones. Quantum dot‑based molecular targeted imaging techniques (which stained pan‑cytokeratin in the cytoplasm green and Ki67 in the cell nucleus yellow or red) were used to investigate the clone formation rate, cell morphology, discrete tendency, and Ki67 expression and distribution in clones. From the cell clone formation assay, the MCF‑7, SW480 and SGC7901 cells were observed to form clones on days 6, 8 and 12 of cell culture, respectively. These three types of cells had heterogeneous morphology, large nuclear:cytoplasmic ratios, and conspicuous pathological mitotic features. The cells at the clone periphery formed multiple pseudopodium. In certain clones, cancer cells at the borderline were separated from the central cell clusters or presented a discrete tendency. With quantum dot‑based molecular targeted imaging techniques, cells with strong Ki67 expression were predominantly shown to be distributed at the clone periphery, or concentrated on one side of the clones. In conclusion, cancer cell clones showed asymmetric growth behavior, and Ki67 was widely expressed in clones of these three cell lines, with strong expression around the clones, or aggregated at one side. Cell clone formation assay based on quantum dots molecular imaging offered a novel method to study the proliferative features of cancer cells, thus providing a further insight into tumor biology.

  18. Quantum dot-based molecular imaging of cancer cell growth using a clone formation assay.

    PubMed

    Geng, Xia-Fei; Fang, Min; Liu, Shao-Ping; Li, Yan

    2016-10-01

    This aim of the present study was to investigate clonal growth behavior and analyze the proliferation characteristics of cancer cells. The MCF‑7 human breast cancer cell line, SW480 human colon cancer cell line and SGC7901 human gastric cancer cell line were selected to investigate the morphology of cell clones. Quantum dot‑based molecular targeted imaging techniques (which stained pan‑cytokeratin in the cytoplasm green and Ki67 in the cell nucleus yellow or red) were used to investigate the clone formation rate, cell morphology, discrete tendency, and Ki67 expression and distribution in clones. From the cell clone formation assay, the MCF‑7, SW480 and SGC7901 cells were observed to form clones on days 6, 8 and 12 of cell culture, respectively. These three types of cells had heterogeneous morphology, large nuclear:cytoplasmic ratios, and conspicuous pathological mitotic features. The cells at the clone periphery formed multiple pseudopodium. In certain clones, cancer cells at the borderline were separated from the central cell clusters or presented a discrete tendency. With quantum dot‑based molecular targeted imaging techniques, cells with strong Ki67 expression were predominantly shown to be distributed at the clone periphery, or concentrated on one side of the clones. In conclusion, cancer cell clones showed asymmetric growth behavior, and Ki67 was widely expressed in clones of these three cell lines, with strong expression around the clones, or aggregated at one side. Cell clone formation assay based on quantum dots molecular imaging offered a novel method to study the proliferative features of cancer cells, thus providing a further insight into tumor biology. PMID:27572664

  19. Mammalian cloning: possibilities and threats.

    PubMed

    Mitalipov, S M; Wolf, D P

    2000-10-01

    The cloning of mammals originated with the production of limited numbers of genetically identical offspring by blastomere separation or embryo splitting. In the past few years, remarkable progress has been reported in cloning by nuclear transfer (NT) with donor nuclei recovered from embryonic, fetal or adult cells. Factors that contribute to the successful reprogramming of the transferred nucleus and the normal term development of the newly reconstructed embryo include the cell cycle stage of both the donor nucleus and recipient cytoplast, the timing of fusion and cytoplast activation, and the source of donor nuclei. The possibility of producing live offspring by somatic cell NT carries potential applications in animal husbandry, biotechnology, transgenic and pharmaceutical production, biomedical research, and the preservation of endangered species. However, the low efficiencies of cloning by NT coupled with high embryonic, fetal and neonatal losses may restrict immediate commercial applications in agriculture. These limitations notwithstanding, the greatest benefits and practical implications of this new technology could be in transplantation medicine and therapeutic cloning.

  20. Clone Poems and the Microcomputer.

    ERIC Educational Resources Information Center

    Irizarry, Estelle

    1989-01-01

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

  1. Healthy ageing of cloned sheep

    PubMed Central

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

    2016-01-01

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

  2. Healthy ageing of cloned sheep.

    PubMed

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

    2016-01-01

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

  3. Healthy ageing of cloned sheep.

    PubMed

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

    2016-01-01

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

  4. Human reproductive cloning: a conflict of liberties.

    PubMed

    Havstad, Joyce C

    2010-02-01

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

  5. Energy values of nine Populus clones

    SciTech Connect

    Strong, T.F.

    1980-01-01

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

  6. Race quickens for the first human clone.

    PubMed

    Gross, M

    2001-04-01

    The dazzling creation of Dolly, the cloned sheep, led many states to legislate against the possibility of using similar technology to create human clones. But for some, this prize is proving too tempting to ignore. Michael Gross reports. PMID:11413008

  7. Genome-wide analysis of methylation in bovine clones by methylated DNA immunoprecipitation (MeDIP).

    PubMed

    Kiefer, Hélène

    2015-01-01

    Methylated DNA immunoprecipitation (MeDIP), when coupled to high-throughput sequencing or microarray hybridization, allows for the identification of methylated loci at a genome-wide scale. Genomic regions affected by incomplete reprogramming after nuclear transfer can potentially be delineated by comparing the MeDIP profiles of bovine clones and non-clones. This chapter presents a MeDIP protocol largely inspired from Mohn and colleagues (Mohn et al., Methods Mol Biol 507:55-64, 2009), with PCR primers specific for cattle, and when possible, overviews of experimental designs adapted to the comparison between clones and non-clones.

  8. Probabilistic cloning of three symmetric states

    SciTech Connect

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

    2010-12-15

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

  9. Phase-covariant quantum cloning of qudits

    SciTech Connect

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

    2003-02-01

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

  10. [Cloning: reproductive medicine or breeding program?].

    PubMed

    Zülicke, F

    1998-01-01

    The presentation of clone-sheep Dolly in February 1997 which was the result of a long and costly research process by Ian Wilmut's team at Roslin Institute near Edinburgh brought world-wide headlines and a continuous debate. But neither cloning and cloning experiments nor the debates about it and the possible application on humans are as new as it is shown in the media. The following article gives some facts and arguments to the field of cloning.

  11. Evaluation of Mutual Drug-Drug Interaction within Geneva Cocktail for Cytochrome P450 Phenotyping using Innovative Dried Blood Sampling Method.

    PubMed

    Bosilkovska, Marija; Samer, Caroline; Déglon, Julien; Thomas, Aurélien; Walder, Bernhard; Desmeules, Jules; Daali, Youssef

    2016-09-01

    Cytochrome P450 (CYP) activity can be assessed using a 'cocktail' phenotyping approach. Recently, we have developed a cocktail (Geneva cocktail) which combines the use of low-dose probes with a low-invasiveness dried blood spots (DBS) sampling technique and a single analytical method for the phenotyping of six major CYP isoforms. We have previously demonstrated that modulation of CYP activity after pre-treatment with CYP inhibitors/inducer could be reliably predicted using Geneva cocktail. To further validate this cocktail, in this study, we have verified whether probe drugs contained in the latter cause mutual drug-drug interactions. In a randomized, four-way, Latin-square crossover study, 30 healthy volunteers received low-dose caffeine, flurbiprofen, omeprazole, dextromethorphan and midazolam (a previously validated combination with no mutual drug-drug interactions); fexofenadine alone; bupropion alone; or all seven drugs simultaneously (Geneva cocktail). Pharmacokinetic profiles of the probe drugs and their metabolites were determined in DBS samples using both conventional micropipette sampling and new microfluidic device allowing for self-sampling. The 90% confidence intervals for the geometric mean ratios of AUC metabolite/AUC probe for CYP probes administered alone or within Geneva cocktail fell within the 0.8-1.25 bioequivalence range indicating the absence of pharmacokinetic interaction. The same result was observed for the chosen phenotyping indices, that is metabolic ratios at 2 hr (CYP1A2, CYP3A) or 3 hr (CYP2B6, CYP2C9, CYP2C19, CYP2D6) post-cocktail administration. DBS sampling could successfully be performed using a new microfluidic device. In conclusion, Geneva cocktail combined with an innovative DBS sampling device can be used routinely as a test for simultaneous CYP phenotyping.

  12. Economical phase-covariant cloning of qudits

    SciTech Connect

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

    2005-04-01

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

  13. Local cloning of arbitrarily entangled multipartite states

    SciTech Connect

    Kay, Alastair; Ericsson, Marie

    2006-01-15

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

  14. Cloning humans, cloning literature: genetics and the imagination deficit.

    PubMed

    Van Dijck, J

    1999-01-01

    After the birth of Dolly, media stories on cloning were replete with references to well-known science fiction plots. This essay criticizes the 'imagination deficit' of scientists and journalists, first by problematizing the uncritical adoption of attentuated science fiction plots in the media coverage of Dolly, and second, by proposing to look at more expansive science fiction novels that carefully examine issues such as uniqueness and identity in relation to the new genetics.

  15. Positional Gene Cloning in Experimental Populations.

    PubMed

    Jagodic, Maja; Stridh, Pernilla

    2016-01-01

    Positional cloning is a technique that identifies a trait-associated gene based on its location in the genome and involves methods such as linkage analysis, association mapping, and bioinformatics. This approach can be used for gene identification even when little is known about the molecular basis of the trait. Vast majority of traits are regulated by multiple genomic loci called quantitative trait loci (QTL). We describe experimental populations and designs that can be used for positional cloning, including backcrosses, intercrosses, and heterogeneous stocks, and advantages and disadvantages of different approaches. Once the phenotype and genotype of each individual in an experimental population have been determined, QTL identification can be accomplished. We describe the statistical tools used to identify the existence, location, and significance of QTLs. These different methods have advantages and disadvantages to consider when selecting the appropriate model to be used, which is briefly discussed.Although the objective of QTL mapping is to identify genomic regions associated with a trait, the ultimate goal is to identify the gene and the genetic variation (which is often quantitative trait nucleotide, QTN) or haplotype that is responsible for the phenotype. By discovering the function of causative variants or haplotypes we can understand the molecular changes that lead to the phenotype. We briefly describe how the genomic sequences can be exploited to identify QTNs and how these can be validated in congenic strains and functionally tested to understand their influence on phenotype expression. PMID:25103675

  16. Computational analysis of small RNA cloning data.

    PubMed

    Berninger, Philipp; Gaidatzis, Dimos; van Nimwegen, Erik; Zavolan, Mihaela

    2008-01-01

    Cloning and sequencing is the method of choice for small regulatory RNA identification. Using deep sequencing technologies one can now obtain up to a billion nucleotides--and tens of millions of small RNAs--from a single library. Careful computational analyses of such libraries enabled the discovery of miRNAs, rasiRNAs, piRNAs, and 21U RNAs. Given the large number of sequences that can be obtained from each individual sample, deep sequencing may soon become an alternative to oligonucleotide microarray technology for mRNA expression profiling. In this report we present the methods that we developed for the annotation and expression profiling of small RNAs obtained through large-scale sequencing. These include a fast algorithm for finding nearly perfect matches of small RNAs in sequence databases, a web-accessible software system for the annotation of small RNA libraries, and a Bayesian method for comparing small RNA expression across samples.

  17. Predators induce cloning in echinoderm larvae.

    PubMed

    Vaughn, Dawn; Strathmann, Richard R

    2008-03-14

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

  18. Probabilistic cloning of three nonorthogonal states

    NASA Astrophysics Data System (ADS)

    Zhang, Wen; Rui, Pinshu; Yang, Qun; Zhao, Yan; Zhang, Ziyun

    2015-04-01

    We study the probabilistic cloning of three nonorthogonal states with equal success probabilities. For simplicity, we assume that the three states belong to a special set. Analytical form of the maximal success probability for probabilistic cloning is calculated. With the maximal success probability, we deduce the explicit form of probabilistic quantum cloning machine. In the case of cloning, we get the unambiguous form of the unitary operation. It is demonstrated that the upper bound for probabilistic quantum cloning machine in (Qiu in J Phys A 35:6931, 2002) can be reached only if the three states are equidistant.

  19. Optimal quantum cloning via spin networks

    SciTech Connect

    Chen Qing; Cheng Jianhua; Wang Kelin; Du Jiangfeng

    2006-09-15

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

  20. No-cloning theorem on quantum logics

    SciTech Connect

    Miyadera, Takayuki; Imai, Hideki

    2009-10-15

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

  1. Therapeutic and reproductive cloning: a critique.

    PubMed

    Bowring, Finn

    2004-01-01

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

  2. Cloning transformations in spin networks without external control

    SciTech Connect

    De Chiara, Gabriele; Fazio, Rosario; Montangero, Simone; Macchiavello, Chiara; Palma, G. Massimo

    2005-07-15

    In this paper 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{yields}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{yields}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.

  3. Implementing phase-covariant cloning in circuit quantum electrodynamics

    NASA Astrophysics Data System (ADS)

    Zhu, Meng-Zheng; Ye, Liu

    2016-10-01

    An efficient scheme is proposed to implement phase-covariant quantum cloning by using a superconducting transmon qubit coupled to a microwave cavity resonator in the strong dispersive limit of circuit quantum electrodynamics (QED). By solving the master equation numerically, we plot the Wigner function and Poisson distribution of the cavity mode after each operation in the cloning transformation sequence according to two logic circuits proposed. The visualizations of the quasi-probability distribution in phase-space for the cavity mode and the occupation probability distribution in the Fock basis enable us to penetrate the evolution process of cavity mode during the phase-covariant cloning (PCC) transformation. With the help of numerical simulation method, we find out that the present cloning machine is not the isotropic model because its output fidelity depends on the polar angle and the azimuthal angle of the initial input state on the Bloch sphere. The fidelity for the actual output clone of the present scheme is slightly smaller than one in the theoretical case. The simulation results are consistent with the theoretical ones. This further corroborates our scheme based on circuit QED can implement efficiently PCC transformation.

  4. Chromatin remodeling in nuclear cloning.

    PubMed

    Wade, Paul A; Kikyo, Nobuaki

    2002-05-01

    Nuclear cloning is a procedure to create new animals by injecting somatic nuclei into unfertilized oocytes. Recent successes in mammalian cloning with differentiated adult nuclei strongly indicate that oocyte cytoplasm contains unidentified remarkable reprogramming activities with the capacity to erase the previous memory of cell differentiation. At the heart of this nuclear reprogramming lies chromatin remodeling as chromatin structure and function define cell differentiation through regulation of the transcriptional activities of the cells. Studies involving the modification of chromatin elements such as selective uptake or release of binding proteins, covalent histone modifications including acetylation and methylation, and DNA methylation should provide significant insight into the molecular mechanisms of nuclear dedifferentiation and redifferentiation in oocyte cytoplasm.

  5. Small Business Innovations

    NASA Technical Reports Server (NTRS)

    1994-01-01

    A Small Business Innovation Research (SBIR) contract resulted in a series of commercially available lasers, which have application in fiber optic communications, difference frequency generation, fiber optic sensing and general laboratory use. Developed under a Small Business Innovation Research (SBIR) contract, the Phase Doppler Particles Analyzer is a non-disruptive, highly accurate laser-based method of determining particle size, number density, trajectory, turbulence and other information about particles passing through a measurement probe volume. The system consists of an optical transmitter and receiver, signal processor and computer with data acquisition and analysis software. A variety of systems are offered for applications including spray characterization for paint, and agricultural and other sprays. The Microsizer, a related product, is used in medical equipment manufacturing and analysis of contained flows. High frequency components and subsystems produced by Millitech Corporation are marketed for both research and commercial use. These systems, which operate in the upper portion of the millimeter wave, resulted from a number of Small Business Innovation Research (SBIR) projects. By developing very high performance mixers and multipliers, the company has advanced the state of the art in sensitive receiver technology. Components are used in receivers and transceivers for monitoring chlorine monoxides, ozone, in plasma characterization and in material properties characterization.

  6. Single cell-derived clones from human adipose stem cells present different immunomodulatory properties.

    PubMed

    Sempere, J M; Martinez-Peinado, P; Arribas, M I; Reig, J A; De La Sen, M L; Zubcoff, J J; Fraga, M F; Fernández, A F; Santana, A; Roche, E

    2014-05-01

    Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single-cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, five single-cell clones were isolated (generally called 1.X and 3.X) from two volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1·10 and 1·22 expressed the lowest amounts, while clones 3·10 and 3·5 expressed more CD105 than the rest and clone 1·7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of interleukin (IL)-6 and low to moderate levels of IL-8. These differences can be explained in part by the distinct methylation profile exhibited by the clones. Furthermore, and after lipopolysaccharide stimulation, clone 3.X produced the highest amounts of proinflammatory cytokines such as IL-1β, while clones 1·10 and 1·22 highly expressed IL-4 and IL-5. In co-culture experiments, clones 1.X are, together, more potent inhibitors than clones 3.X for proliferation of total, CD3(+) T, CD4(+) T and CD8(+) T lymphocytes and natural killer (NK) cells. The results of this work indicate that the adipose stem cell population is heterogeneous in cytokine production profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies.

  7. Telomere elongation facilitated by trichostatin a in cloned embryos and pigs by somatic cell nuclear transfer.

    PubMed

    Kong, Qingran; Ji, Guangzhen; Xie, Bingteng; Li, Jingyu; Mao, Jian; Wang, Juan; Liu, Shichao; Liu, Lin; Liu, Zhonghua

    2014-06-01

    Telomere attrition and genomic instability are associated with organism aging. Concerns still exist regarding telomere length resetting in cloned embryos and ntES cells, and possibilities of premature aging of cloned animals achieved by somatic cell nuclear transfer (SCNT). Trichostatin A (TSA), a histone deacetylase inhibitor, effectively improves the developmental competence of cloned embryos and animals, and recently contributes to successful generation of human ntES cells by SCNT. To test the function of TSA on resetting telomere length, we analyzed telomeres in cloned blastocysts and pigs following treatment of SCNT embryos with TSA. Here, we show that telomeres of cloned pigs generated by standard SCNT methods are not effectively restored, compared with those of donor cells, however TSA significantly increases telomere lengths in cloned pigs. Telomeres elongate in cloned porcine embryos during early cleavage from one-cell to four-cell stages. Notably, TSA facilitates telomere lengthening of cloned embryos mainly at morula-blastocyst stages. Knockdown of pTert by shRNA in donor cells reduces telomerase activity in cloned blastocysts but does not abrogate telomere elongation in the TSA-treated embryos (p > 0.05). However, genes associated with recombination or telomerase-independent mechanism of alternative lengthening of telomeres (ALT) Rad50 and BLM show increased expression in TSA-treated embryos. These data suggest that TSA may promote telomere elongation of cloned porcine embryos by ALT. Together, TSA can elongate telomeres in cloned embryos and piglets, and this could be one of the mechanisms underlying improved development of cloned embryos and animals treated with TSA. PMID:24510582

  8. Single cell-derived clones from human adipose stem cells present different immunomodulatory properties

    PubMed Central

    Sempere, J M; Martinez-Peinado, P; Arribas, M I; Reig, J A; De La Sen, M L; Zubcoff, J J; Fraga, M F; Fernández, A F; Santana, A; Roche, E

    2014-01-01

    Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single-cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, five single-cell clones were isolated (generally called 1.X and 3.X) from two volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1·10 and 1·22 expressed the lowest amounts, while clones 3·10 and 3·5 expressed more CD105 than the rest and clone 1·7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of interleukin (IL)-6 and low to moderate levels of IL-8. These differences can be explained in part by the distinct methylation profile exhibited by the clones. Furthermore, and after lipopolysaccharide stimulation, clone 3.X produced the highest amounts of proinflammatory cytokines such as IL-1β, while clones 1·10 and 1·22 highly expressed IL-4 and IL-5. In co-culture experiments, clones 1.X are, together, more potent inhibitors than clones 3.X for proliferation of total, CD3+T, CD4+T and CD8+T lymphocytes and natural killer (NK) cells. The results of this work indicate that the adipose stem cell population is heterogeneous in cytokine production profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies. PMID:24666184

  9. The Need for Innovative Methods of Teaching and Learning Chemistry in Higher Education--Reflections from a Project of the European Chemistry Thematic Network

    ERIC Educational Resources Information Center

    Eilks, Ingo; Byers, Bill

    2010-01-01

    This paper summarizes the work and conclusions of a working group established by the European Chemistry Thematic Network (ECTN). The aim of the working group was to identify potential areas for innovative approaches to the teaching and learning of chemistry in Higher Education, and to survey good practice throughout the EU. The paper starts by…

  10. Clone-preventive technique that features magnetic microfibers and cryptography

    NASA Astrophysics Data System (ADS)

    Matsumoto, Hiroyuki; Suzuki, Keiichi; Matsumoto, Tsutomu

    1998-04-01

    We have used the term 'clone' to refer to those things which are produced by methods such as counterfeiting, alteration, duplication or simulation. To satisfy the requirements of secure and low-cost techniques for preventing card fraud, we have recently developed a clone preventive system called 'FibeCrypt (Fiber Cryptosystem)' which utilizes physical characteristics. Each card has a canonical domain (i.e. a distinctive part), similar to fingerprints as the biometric measurement, made up of magnetic micro-fibers scattered randomly inside. We have applied cryptosystems to the system. FibeCrypt examines and authenticates the unique pattern of the canonical domain using pre-stored reference data and a digital signature. In our paper, the schemes and the features of this system are described in detail. The results of our examinations show the accuracy of authentication of the system. We conclude that this authentication technique which utilizes physical characteristics can be very effective for clone prevention in various fields.

  11. Assembly of Multigene Constructs Using Golden Gate Cloning.

    PubMed

    Marillonnet, Sylvestre; Werner, Stefan

    2015-01-01

    Efficient DNA assembly methods are required for synthetic biology. Standardization of DNA parts is an essential element that not only facilitates reuse of the same parts for various constructs but also allows standardization of the assembly strategy. We provide here a protocol for assembly of multigene constructs from standard biological parts using the modular cloning system MoClo. Making constructs using this system requires to first define the structure of the final construct and to identify all basic parts and vectors required for the construction strategy. The cloning strategy is in large part determined by the structure of the final construct, which is then made using a series of one-pot Golden Gate cloning reactions. PMID:26082229

  12. Cloning Expeditions: Risky but Rewarding

    PubMed Central

    2013-01-01

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

  13. Cloning expeditions: risky but rewarding.

    PubMed

    Lodish, Harvey

    2013-12-01

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

  14. Production of Cloned Mice by Nuclear Transfer of Cumulus Cells

    PubMed Central

    Kurd, Soleiman; Zarei, Mohammad Ali; Fathi, Fardin; Ghadimi, Tayyeb; Hakhamaneshi, Mohammad Saeed; Jalili, Ali

    2013-01-01

    Background Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells (NT) into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator. Methods B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice. Results Of 367 oocytes collected, 131 (69%) developed into 2-cell stage embryos. Of these, 5 (1%) live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring. Conclusion For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit. PMID:23919122

  15. Expression cloning of human B cell immunoglobulins.

    PubMed

    Wardemann, Hedda; Kofer, Juliane

    2013-01-01

    The majority of lymphomas originate from B cells at the germinal center stage or beyond. Preferential selection of B cell clones by a limited set of antigens has been suggested to drive lymphoma development. However, little is known about the specificity of the antibodies expressed by lymphoma cells, and the role of antibody-specificity in lymphomagenesis remains elusive. Here, we describe a strategy to characterize the antibody reactivity of human B cells. The approach allows the unbiased characterization of the human antibody repertoire on a single cell level through the generation of recombinant monoclonal antibodies from single primary human B cells of defined origin. This protocol offers a detailed description of the method starting from the flow cytometric isolation of single human B cells, to the RT-PCR-based amplification of the expressed Igh, Igκ, and Igλ chain genes, and Ig gene expression vector cloning for the in vitro production of monoclonal antibodies. The strategy may be used to obtain information on the clonal evolution of B cell lymphomas by single cell Ig gene sequencing and on the antibody reactivity of human lymphoma B cells.

  16. Spinning the Innovation and Entrepreneurship Mindset: A Modern Physics Approach

    NASA Astrophysics Data System (ADS)

    Roughani, Bahram

    2010-04-01

    Topics in Modern Physics course from relativity to quantum mechanics were examined in the context of innovation as part of the recent Kettering University program on ``Entrepreneurship Across Curriculum-EAC.'' The main goals were (a) to introduce innovation and entrepreneurship without eliminating any topics from this course, (b) to use EAC as a vehicle for intentional education that produces graduates with innovative mindsets, (c) to enrich the students learning experience aligned with the desired educational outcomes, and (d) to highlight the impact of scientific innovation in the society, while encouraging students to re-think how entrepreneurship mindset could maximize their impact in the society through innovation. Ideas such as principles behind innovation and innovative ideas, disciplines of innovations, formation of innovation teams, and effective methods for analyzing innovative value propositions were introduced in this course. Most of the implementation were achieved through out of class activities, and communicated through in class presentations, papers or weekly laboratory reports.

  17. An Innovative Collagen-Based Cell-Printing Method for Obtaining Human Adipose Stem Cell-Laden Structures Consisting of Core-Sheath Structures for Tissue Engineering.

    PubMed

    Yeo, MyungGu; Lee, Ji-Seon; Chun, Wook; Kim, Geun Hyung

    2016-04-11

    Three-dimensional (3D) cell printing processes have been used widely in various tissue engineering applications due to the efficient embedding of living cells in appropriately designed micro- or macro-structures. However, there are several issues to overcome, such as the limited choice of bioinks and tailor-made fabricating strategies. Here, we suggest a new, innovative cell-printing process, supplemented with a core-sheath nozzle and an aerosol cross-linking method, to obtain multilayered cell-laden mesh structure and a newly considered collagen-based cell-laden bioink. To obtain a mechanically and biologically enhanced cell-laden structure, we used collagen-bioink in the core region, and also used pure alginate in the sheath region to protect the cells in the collagen during the printing and cross-linking process and support the 3D cell-laden mesh structure. To achieve the most appropriate conditions for fabricating cell-embedded cylindrical core-sheath struts, various processing conditions, including weight fractions of the cross-linking agent and pneumatic pressure in the core region, were tested. The fabricated 3D MG63-laden mesh structure showed significantly higher cell viability (92 ± 3%) compared with that (83 ± 4%) of the control, obtained using a general alginate-based cell-printing process. To expand the feasibility to stem cell-embedded structures, we fabricated a cell-laden mesh structure consisting of core (cell-laden collagen)/sheath (pure alginate) using human adipose stem cells (hASCs). Using the selected processing conditions, we could achieve a stable 3D hASC-laden mesh structure. The fabricated cell-laden 3D core-sheath structure exhibited outstanding cell viability (91%) compared to that (83%) of an alginate-based hASC-laden mesh structure (control), and more efficient hepatogenic differentiations (albumin: ∼ 1.7-fold, TDO-2: ∼ 7.6-fold) were observed versus the control. The selection of collagen-bioink and the new printing strategy

  18. An Innovative Collagen-Based Cell-Printing Method for Obtaining Human Adipose Stem Cell-Laden Structures Consisting of Core-Sheath Structures for Tissue Engineering.

    PubMed

    Yeo, MyungGu; Lee, Ji-Seon; Chun, Wook; Kim, Geun Hyung

    2016-04-11

    Three-dimensional (3D) cell printing processes have been used widely in various tissue engineering applications due to the efficient embedding of living cells in appropriately designed micro- or macro-structures. However, there are several issues to overcome, such as the limited choice of bioinks and tailor-made fabricating strategies. Here, we suggest a new, innovative cell-printing process, supplemented with a core-sheath nozzle and an aerosol cross-linking method, to obtain multilayered cell-laden mesh structure and a newly considered collagen-based cell-laden bioink. To obtain a mechanically and biologically enhanced cell-laden structure, we used collagen-bioink in the core region, and also used pure alginate in the sheath region to protect the cells in the collagen during the printing and cross-linking process and support the 3D cell-laden mesh structure. To achieve the most appropriate conditions for fabricating cell-embedded cylindrical core-sheath struts, various processing conditions, including weight fractions of the cross-linking agent and pneumatic pressure in the core region, were tested. The fabricated 3D MG63-laden mesh structure showed significantly higher cell viability (92 ± 3%) compared with that (83 ± 4%) of the control, obtained using a general alginate-based cell-printing process. To expand the feasibility to stem cell-embedded structures, we fabricated a cell-laden mesh structure consisting of core (cell-laden collagen)/sheath (pure alginate) using human adipose stem cells (hASCs). Using the selected processing conditions, we could achieve a stable 3D hASC-laden mesh structure. The fabricated cell-laden 3D core-sheath structure exhibited outstanding cell viability (91%) compared to that (83%) of an alginate-based hASC-laden mesh structure (control), and more efficient hepatogenic differentiations (albumin: ∼ 1.7-fold, TDO-2: ∼ 7.6-fold) were observed versus the control. The selection of collagen-bioink and the new printing strategy

  19. Implementing innovations in health care settings.

    PubMed

    MacDonald, V; Muir, J

    1996-10-01

    Innovations in health care settings are occurring at an unprecedented rate. New methods and ideas include computerized pumps, computer systems for documentation and communication, and alternative approaches to patient care. To be successfully adopted by nurses, innovations require well-planned administrative, educational and clinical support. A multi-agency research study has revealed factors that should be considered when planning innovations in health care settings. PMID:9118058

  20. Bricks and blueprints: methods and standards for DNA assembly.

    PubMed

    Casini, Arturo; Storch, Marko; Baldwin, Geoffrey S; Ellis, Tom

    2015-09-01

    DNA assembly is a key part of constructing gene expression systems and even whole chromosomes. In the past decade, a plethora of powerful new DNA assembly methods - including Gibson Assembly, Golden Gate and ligase cycling reaction (LCR) - have been developed. In this Innovation article, we discuss these methods as well as standards such as the modular cloning (MoClo) system, GoldenBraid, modular overlap-directed assembly with linkers (MODAL) and PaperClip, which have been developed to facilitate a streamlined assembly workflow, to aid the exchange of material between research groups and to create modular reusable DNA parts.

  1. Accelerated Innovation Pilot

    NASA Technical Reports Server (NTRS)

    Davis, Jeffrey

    2012-01-01

    Opportunities: I. Engage NASA team (examples) a) Research and technology calls . provide suggestions to AES, HRP, OCT. b) Use NASA@Work to solicit other ideas; (possibly before R+D calls). II. Stimulate collaboration (examples) a) NHHPC. b) Wharton Mack Center for Technological Innovation (Feb 2013). c) International ] DLR ] :envihab (July 2013). d) Accelerated research models . NSF, Myelin Repair Foundation. III. Engage public Prizes (open platform: InnoCentive, yet2.com, NTL; Rice Business Plan, etc.) IV. Use same methods to engage STEM.

  2. Innovations in bereavement education.

    PubMed

    Wright, Patricia Moyle

    2011-08-01

    Advanced practice nursing students provide care for clients and families in numerous settings where they will encounter end-of-life issues. Thus, graduate nursing education should include information on current trends in thanatology, such as the debate over the proposed complicated grief criteria and the paradigmatic shift toward evidence-based grief theory. In this article, an innovative approach to teaching bereavement content to graduate nursing students during a 3-hour class is presented. The assignments were developed specifically for adult learners with clinical experience. Students' responses to the learning activities and recommendations for modifications of the teaching methods are presented.

  3. Innovation in healthcare: a systematic review of recent research.

    PubMed

    Länsisalmi, Hannakaisa; Kivimäki, Mika; Aalto, Pirjo; Ruoranen, Raija

    2006-01-01

    Research on innovations in healthcare organizations published between 1994 and 2004 are here reviewed and summarized. The majority of the 31 identified studies dealt with the adoption of innovations and new practices and were cross-sectional designs applying quantitative methods, or multiple case studies applying qualitative methods. Five pathways for future research are recommended: (a) Multilevel approaches studying innovation simultaneously on individual, group, and organizational levels; (b) a combination of quantitative and qualitative data; (c) use of longitudinal designs (innovation both as the dependent and independent variable); (d) application of experimental designs in interventions; and (e) exploration of innovation generation and structural innovations.

  4. Agro-economic impact of cattle cloning.

    PubMed

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

    2004-01-01

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

  5. Agro-economic impact of cattle cloning.

    PubMed

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

    2004-01-01

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

  6. A rapid screening procedure for the identification of high-titer retrovirus packaging clones.

    PubMed

    Murdoch, B; Pereira, D S; Wu, X; Dick, J E; Ellis, J

    1997-07-01

    We have developed a viral RNA (vRNA) dot blot assay for rapid identification of high-titer retrovirus vector production by packaging cell clones. The procedure employs Trizol LS reagent to purify vRNA from packaging cell supernatants, a sensitive dot blot assay, and Phosphorlmager technology to quantify packaged viral genomes in 2 days. Experiments performed on viral supernatants of known biological titer demonstrated that the vRNA dot blot assay was extremely sensitive and that dot intensity correlated directly with viral titer. It is often necessary to analyze approximately 100 virus producing cell clones, making this method useful as a rapid screen to identify the highest virus producing clones. The vRNA dot blot assay consistently identified a subset of candidate high-titer producer cell clones. In three independent screens the supernatant with the highest biological titer was produced by one of the previously defined candidate high-titer producer clones. Our procedure greatly facilitates virus titration by: (1) rapidly eliminating the vast majority of low-titer producer cell clones; (2) accurately identifying the subset of candidate high-titer producer clones for further biological titration and assessment of the proviral genomic structure; and (3) reducing laborious tissue culture manipulations to a minimum. Furthermore, the reliance of this method on molecular detection makes it ideally suited for the isolation of high-titer clones lacking a drug selection marker.

  7. The innovative moments coding system and the assimilation of problematic experiences scale: a case study comparing two methods to track change in psychotherapy.

    PubMed

    Gonçalves, Miguel M; Gabalda, Isabel Caro; Ribeiro, António P; Pinheiro, PatrÍcia; Borges, Raquel; Sousa, Inês; Stiles, William B

    2014-01-01

    The Assimilation of Problematic Experiences Scale (APES) and the Innovative Moments Coding System were applied to transcripts of a successful case of linguistic therapy of evaluation independently by different research groups. Assimilation theory and research suggest that higher APES scores reflect therapeutic gains, with a level of approximately 4.0 separating good from poor outcome cases. The innovative moments (IMs) model suggests that IMs classified as reconceptualization and performing change occur mainly in good outcome cases, whereas action, reflection and protest occur in both good and poor outcome cases. Passages coded as reconceptualization and performing change were rare in this case, but 100% of them were rated at or above APES 4. By contrast, 63% passages coded as action, reflection or protest were rated below APES 4 (Chi-square = 28.62, p < .001). Implications for research are discussed. PMID:24099105

  8. Genetic analysis of yeast strains lacking negative feedback control: a one-step method for positive selection and cloning of carbamoylphosphate synthetase-aspartate transcarbamoylase mutants unable to respond to UTP.

    PubMed

    Jaquet, L; Lollier, M; Souciet, J L; Potier, S

    1993-10-01

    We have undertaken an in vivo genetic approach to the analysis of negative feedback control by uridine triphosphate (UTP) of the yeast carbamoylphosphate synthetase-aspartate transcarbamoylase multifunctional protein (CPSase-ATCase). Using an analog of uracil, 5-fluorouracil, we have constructed a screening system leading, in one step, to selection and cloning of a functional aspartate transcarbamoylase that is defective in negative feedback control by UTP. Due to the nature of the screen, spontaneous or UV-induced mutants could be recovered. Well-characterized cloned mutants have been sequenced and reveal one or two modifications in single codons leading to single amino acid replacements. These amino acid changes occurred either in the CPSase or ATCase domains, abolishing their sensitivity to regulation but not their catalytic activities. Hence the regulatory and catalytic sites are distinct. With the same screening system, it may also be possible to enlarge the scope of the molecular study of the feedback processes to include equivalent proteins in fungi as well as higher eukaryotes. PMID:8232215

  9. Cloning

    MedlinePlus

    ... mammals. These twins are produced when a fertilized egg splits, creating two or more embryos that carry ... of the donor animal's somatic cell into an egg cell, or oocyte, that has had its own ...

  10. Unified universal quantum cloning machine and fidelities

    SciTech Connect

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

    2011-09-15

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

  11. Nuclear transfer technology in mammalian cloning.

    PubMed

    Wolf, D P; Mitalipov, S; Norgren, R B

    2001-01-01

    The past several years have witnessed remarkable progress in mammalian cloning using nuclear transfer (NT). Until 1997 and the announcement of the successful cloning of sheep from adult mammary gland or fetal fibroblast cells, our working assumption was that cloning by NT could only be accomplished with relatively undifferentiated embryonic cells. Indeed, live offspring were first produced by NT over 15 years ago from totipotent, embryonic blastomeres derived from early cleavage-stage embryos. However, once begun, the progression to somatic cell cloning or NT employing differentiated cells as the source of donor nuclei was meteoric, initially involving differentiated embryonic cell cultures in sheep in 1996 and quickly thereafter, fetal or adult somatic cells in sheep, cow, mouse, goat, and pig. Several recent reviews provide a background for and discussion of these successes. Here we will focus on the potential uses of reproductive cloning along with recent activities in the field and a discussion concerning current interests in human reproductive and therapeutic cloning.

  12. No human cloning: a social ethics perspective.

    PubMed

    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. PMID:12650145

  13. Cloning: pathways to a pluripotent future.

    PubMed

    McLaren, A

    2000-06-01

    In this month's essay, Anne McLaren traces the winding and pitted pathways that connect the early days of the cell theory of biology in the 1830s to the new and unfolding era of cloning science and technology that came to worldwide attention in 1997 with the announcement of the birth of Dolly, the Scottish cloned sheep. The possibilities, including the potential for new medical treatments and perhaps even human cloning, are fantastic ... and ethically charged.

  14. Telomeres and the ethics of human cloning.

    PubMed

    Allhoff, Fritz

    2004-01-01

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

  15. Cloning: pathways to a pluripotent future.

    PubMed

    McLaren, A

    2000-06-01

    In this month's essay, Anne McLaren traces the winding and pitted pathways that connect the early days of the cell theory of biology in the 1830s to the new and unfolding era of cloning science and technology that came to worldwide attention in 1997 with the announcement of the birth of Dolly, the Scottish cloned sheep. The possibilities, including the potential for new medical treatments and perhaps even human cloning, are fantastic ... and ethically charged. PMID:10877698

  16. Cloning of spin-coherent states

    SciTech Connect

    Demkowicz-Dobrzanski, Rafal; Kus, Marek; Wodkiewicz, Krzysztof

    2004-01-01

    We consider optimal cloning of the spin coherent states in Hilbert spaces of different dimensionality d. We give explicit form of optimal cloning transformation for spin coherent states in the three-dimensional space, analytical results for the fidelity of the optimal cloning in d=3 and d=4 as well as numerical results for higher dimensions. In the low-dimensional case we construct the corresponding completely positive maps and exhibit their structure with the help of Jamiolkowski isomorphism. This allows us to formulate some conjectures about the form of optimal coherent cloning completely positive maps in arbitrary dimension.

  17. Quantum cloning disturbed by thermal Davies environment

    NASA Astrophysics Data System (ADS)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-06-01

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

  18. Species-specific challenges in dog cloning.

    PubMed

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

    2012-12-01

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

  19. Human cloning: Eastern Mediterranean Region perspective.

    PubMed

    Abdur Rab, M; Khayat, M H

    2006-01-01

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

  20. [Human cloning: judicial and legislative framework].

    PubMed

    Shenfield, F

    2000-09-01

    The application, either theoretical or fantastical, of the somatic cloning which resulted in the birth of Dolly the sheep to human reproduction has led to international uproar. A paragraph specifically banning reproductive cloning (defined as "any intervention seeking to produce genetically identical human individuals in the sense of individuals sharing the same nuclear gene set") was added to the Council of Europe Convention on Human Rights and Biomedicine in January 1998. In several responses both at national and international level, reproductive cloning is banned either directly or indirectly. The challenge for the future is the consideration of integrating therapeutic cloning in legislations both nationally and in international declarations.

  1. No end in sight to cloning debate.

    PubMed

    Graumann, Sigrid; Poltermann, Andreas

    2005-01-01

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

  2. Cloning by somatic cell nuclear transfer.

    PubMed

    Fulka, J; First, N L; Loi, P; Moor, R M

    1998-10-01

    The birth of the first cloned mammals, produced by the introduction of somatic cell nuclei into enucleated oocytes, was an impressive and surprising development. Although the ethical debate has been intense, the important scientific questions raised by this work have been inadequately discussed and are still unresolved. In this essay we address three questions about nuclear transplantation in the eggs of mice and domestic animals. First, why were the recent experiments on somatic cell cloning successful, when so many others have failed? Second, were these exceptional cases, or is somatic cloning now open to all? Third, what are the future possibilities for increasing the efficiency and wider applicability of the cloning process?

  3. [Human cloning: judicial and legislative framework].

    PubMed

    Shenfield, F

    2000-09-01

    The application, either theoretical or fantastical, of the somatic cloning which resulted in the birth of Dolly the sheep to human reproduction has led to international uproar. A paragraph specifically banning reproductive cloning (defined as "any intervention seeking to produce genetically identical human individuals in the sense of individuals sharing the same nuclear gene set") was added to the Council of Europe Convention on Human Rights and Biomedicine in January 1998. In several responses both at national and international level, reproductive cloning is banned either directly or indirectly. The challenge for the future is the consideration of integrating therapeutic cloning in legislations both nationally and in international declarations. PMID:11075502

  4. Innovations and Innovation Processes in Higher Education.

    ERIC Educational Resources Information Center

    Berg, Barbro; Ostergren, Bertil

    Included in this report are seven comparative case studies of innovation processes in higher education in Sweden. The purpose of the study is to chart dysfunctions in the system in planning for change, and also to gain more knowledge of the organization by studying it when it has been "disturbed" through introduction of an innovative process. The…

  5. Optimization of embryo culture conditions for increasing efficiency of cloning in buffalo (Bubalus bubalis) and generation of transgenic embryos via cloning.

    PubMed

    Wadhwa, Neerja; Kunj, Neetu; Tiwari, Shuchita; Saraiya, Megha; Majumdar, Subeer S

    2009-09-01

    Cloning in bovine species is marred by low efficiency of blastocyst formation. Any increase in the efficiency of blastocyst formation upon nuclear transfer will greatly enhance the efficiency of cloning. In the present study, the effect of various media, protein sources, and growth factors on the development of cloned buffalo embryos was evaluated. Among various combinations tested, culture of cloned embryos in TCM-199 media on the feeder layer of Buffalo Oviductal Epithelial Cells (BOEC) in the presence of bovine serum albumin-free fatty acid (BSA-FFA) and leukemia inhibitory factor (LIF) provided most suitable environment for efficient development of cloned blastocysts. Under these conditions, we achieved a blastocyst formation rate of 43%, which is better than those reported previously. Because preimplantation embryonic development, in vivo, occurs in an environment of oviductal cells, the blastocysts generated by this method may presumably be more suitable for implantation and further development. Additionally, we generated green blastocysts from enucleated oocytes by transfer of nuclei from cells transfected with EGFP transgene, showing possibility of transgenesis via cloning in this species. To our knowledge, this is the first report regarding the production of transgenic cloned buffalo embryos and their developmental competence with respect to various media, cocultures, and supplements.

  6. Cloning of expansin genes in ramie (Boehmeria nivea L.) based on universal fast walking.

    PubMed

    Chen, Jie; Dai, Lunjin; Wang, Bo; Liu, Lijun; Peng, Dingxiang

    2015-09-10

    Gene cloning is the first step to study the expression profiles and functions of a particular gene; considerable cloning methods have been developed. Expansin, thought to involve in the cell-wall modification events, was not cloned in ramie (Boehmeria nivea L.), which is one of the most important bast fiber crops with little conducted molecular research, especially on its fiber development. Studying the expansin gene family will uncover its possible relationship with ramie fiber development and other growth events. As a result, five expansin genes were cloned with full-length and their sequence information was investigated. Additionally, the phylogenetic analysis was conducted, which suggested that the cloned genes belong to the α-subfamily, and these genes expressed differently during ramie fiber developmental process. In this study, we aimed to apply a strategy for cloning novel full-length genes from genomic DNA of ramie, based on using degenerate primers, touchdown polymerase chain reaction and universal fast walking protocols. By cloning five full-length expansin genes, we believe the polymerase chain reaction-based gene cloning strategy could be applied to general gene studies in ramie and other crops.

  7. Innovation and STEM Schools

    ERIC Educational Resources Information Center

    Roberts, Julia Link

    2015-01-01

    How do schools with a focus on science, technology, engineering, and mathematics (STEM) fit in with state goals to increase innovation and to boost the economy? This article briefly discusses how educators can encourage creativity and innovation.

  8. Innovations: Scientific, Technological, and Social.

    ERIC Educational Resources Information Center

    Gabor, Dennis

    Dr. Gabor, the inventor of holography (lenseless photography), defines "innovation" as a methodical creation of the human spirit, a novelty that once created can be usefully and repeatedly applied. He describes and evaluates 100 important technological and biological inventions that can probably be expected within the next 50 years. He also…

  9. Mitochondrial DNA heteroplasmy in cloned cattle produced by fetal and adult cell cloning.

    PubMed

    Steinborn, R; Schinogl, P; Zakhartchenko, V; Achmann, R; Schernthaner, W; Stojkovic, M; Wolf, E; Müller, M; Brem, G

    2000-07-01

    Mammals have been cloned from adult donor cells. Here we report the first cases of mitochondrial DNA (mtDNA) heteroplasmy in adult mammalian clones generated from fetal and adult donor cells. The heteroplasmic clones included a healthy cattle equivalent of the sheep Dolly, for which a lack of heteroplasmy was reported.

  10. Mitochondrial DNA heteroplasmy in cloned cattle produced by fetal and adult cell cloning.

    PubMed

    Steinborn, R; Schinogl, P; Zakhartchenko, V; Achmann, R; Schernthaner, W; Stojkovic, M; Wolf, E; Müller, M; Brem, G

    2000-07-01

    Mammals have been cloned from adult donor cells. Here we report the first cases of mitochondrial DNA (mtDNA) heteroplasmy in adult mammalian clones generated from fetal and adult donor cells. The heteroplasmic clones included a healthy cattle equivalent of the sheep Dolly, for which a lack of heteroplasmy was reported. PMID:10888867

  11. China's Innovation Paradox

    ERIC Educational Resources Information Center

    Chan, Jeremy

    2015-01-01

    China aims to become an innovation-led nation by 2020, but its leadership is generally sceptical--and oftentimes hostile--to the market forces, open exchange of ideas, and creative destruction that have unlocked innovation in other countries. Instead, Beijing hopes to promote innovation in China through a massive expansion in higher education,…

  12. The Impact of Innovation

    ERIC Educational Resources Information Center

    O'Banion, Terry; Weidner, Laura; Wilson, Cynthia

    2012-01-01

    With funding from MetLife Foundation, the League for Innovation in the Community College engaged in a yearlong study in 2009 of the nature of innovation in the community college. Using recipients of the League's Innovation of the Year Award at 19 community colleges during the period from 1999 through 2008 as a data set, the authors used document…

  13. Who Are the Innovators?

    ERIC Educational Resources Information Center

    O'Banion, Terry; Weidner, Laura; Wilson, Cynthia

    2010-01-01

    As we approach the second decade of the new millennium, there is a renaissance of innovation in education, a resurgence of interest and experimentation that begs for analysis and review. To that end, the League for Innovation in the Community College proposed to conduct a national study on the nature of innovation in the community college using…

  14. JPL Innovation Foundry

    NASA Technical Reports Server (NTRS)

    Sherwood, Brent; McCleese, Daniel J.

    2012-01-01

    NASA supports the community of mission principal investigators by helping them ideate, mature, and propose concepts for new missions. As NASA's Federally Funded Research and Development Center (FFRDC), JPL is a primary resource for providing this service. The environmental context for the formulation lifecycle evolves continuously. Contemporary trends include: more competitors; more-complex mission ideas; scarcer formulation resources; and higher standards for technical evaluation. Derived requirements for formulation support include: stable, clear, reliable methods tailored for each stage of the formulation lifecycle; on-demand access to standout technical and programmatic subject-matter experts; optimized, outfitted facilities; smart access to learning embodied in a vast oeuvre of prior formulation work; hands-on method coaching. JPL has retooled its provision of integrated formulation lifecycle support to PIs, teams, and program offices in response to this need. This mission formulation enterprise is the JPL Innovation Foundry.

  15. Innovations in national nutrition surveys.

    PubMed

    Stephen, Alison M; Mak, Tsz Ning; Fitt, Emily; Nicholson, Sonja; Roberts, Caireen; Sommerville, Jill

    2013-02-01

    The aim of this paper is to describe innovations taking place in national nutrition surveys in the UK and the challenges of undertaking innovations in such settings. National nutrition surveys must be representative of the overall population in characteristics such as socio-economic circumstances, age, sex and region. High response rates are critical. Dietary assessment innovations must therefore be suitable for all types of individuals, from the very young to the very old, for variable literacy and/or technical skills, different ethnic backgrounds and life circumstances, such as multiple carers and frequent travel. At the same time, national surveys need details on foods consumed. Current advances in dietary assessment use either technological innovations or simplified methods; neither lend themselves to national surveys. The National Diet and Nutrition Survey (NDNS) rolling programme, and the Diet and Nutrition Survey of Infants and Young Children (DNSIYC), currently use the 4-d estimated diary, a compromise for detail and respondent burden. Collection of food packaging enables identification of specific products. Providing space for location of eating, others eating, the television being on and eating at a table, adds to eating context information. Disaggregation of mixed dishes enables determination of true intakes of meat and fruit and vegetables. Measurement of nutritional status requires blood sampling and processing in DNSIYC clinics throughout the country and mobile units were used to optimise response. Hence, innovations in national surveys can and are being made but must take into account the paramount concerns of detail and response rate.

  16. Particle infectivity of HIV-1 full-length genome infectious molecular clones in a subtype C heterosexual transmission pair following high fidelity amplification and unbiased cloning

    SciTech Connect

    Deymier, Martin J.; Claiborne, Daniel T.; Ende, Zachary; Ratner, Hannah K.; Kilembe, William; Hunter, Eric

    2014-11-15

    The high genetic diversity of HIV-1 impedes high throughput, large-scale sequencing and full-length genome cloning by common restriction enzyme based methods. Applying novel methods that employ a high-fidelity polymerase for amplification and an unbiased fusion-based cloning strategy, we have generated several HIV-1 full-length genome infectious molecular clones from an epidemiologically linked transmission pair. These clones represent the transmitted/founder virus and phylogenetically diverse non-transmitted variants from the chronically infected individual's diverse quasispecies near the time of transmission. We demonstrate that, using this approach, PCR-induced mutations in full-length clones derived from their cognate single genome amplicons are rare. Furthermore, all eight non-transmitted genomes tested produced functional virus with a range of infectivities, belying the previous assumption that a majority of circulating viruses in chronic HIV-1 infection are defective. Thus, these methods provide important tools to update protocols in molecular biology that can be universally applied to the study of human viral pathogens. - Highlights: • Our novel methodology demonstrates accurate amplification and cloning of full-length HIV-1 genomes. • A majority of plasma derived HIV variants from a chronically infected individual are infectious. • The transmitted/founder was more infectious than the majority of the variants from the chronically infected donor.

  17. Innovation and design approaches within prospective ergonomics.

    PubMed

    Liem, André; Brangier, Eric

    2012-01-01

    In this conceptual article the topic of "Prospective Ergonomics" will be discussed within the context of innovation, design thinking and design processes & methods. Design thinking is essentially a human-centred innovation process that emphasises observation, collaboration, interpretation, visualisation of ideas, rapid concept prototyping and concurrent business analysis, which ultimately influences innovation and business strategy. The objective of this project is to develop a roadmap for innovation, involving consumers, designers and business people in an integrative process, which can be applied to product, service and business design. A theoretical structure comprising of Innovation perspectives (1), Worldviews supported by rationalist-historicist and empirical-idealistic dimensions (2) and Models of "design" reasoning (3) precedes the development and classification of existing methods as well as the introduction of new ones.

  18. Innovation and design approaches within prospective ergonomics.

    PubMed

    Liem, André; Brangier, Eric

    2012-01-01

    In this conceptual article the topic of "Prospective Ergonomics" will be discussed within the context of innovation, design thinking and design processes & methods. Design thinking is essentially a human-centred innovation process that emphasises observation, collaboration, interpretation, visualisation of ideas, rapid concept prototyping and concurrent business analysis, which ultimately influences innovation and business strategy. The objective of this project is to develop a roadmap for innovation, involving consumers, designers and business people in an integrative process, which can be applied to product, service and business design. A theoretical structure comprising of Innovation perspectives (1), Worldviews supported by rationalist-historicist and empirical-idealistic dimensions (2) and Models of "design" reasoning (3) precedes the development and classification of existing methods as well as the introduction of new ones. PMID:22317532

  19. Molecular cloning of gluconobacter oxydans DSM 2003 xylitol dehydrogenase gene

    PubMed Central

    Sadeghi, H. Mir Mohammad; Ahmadi, R.; Aghaabdollahian, S.; Mofid, M.R.; Ghaemi, Y.; Abedi, D.

    2011-01-01

    Due to the widespread applications of xylitol dehydrogenase, an enzyme used for the production of xylitol, the present study was designed for the cloning of xylitol dehydrogenase gene from Glcunobacter oxydans DSM 2003. After extraction of genomic DNA from this bacterium, xylitol dehydrogenase gene was replicated using polymerase chain reaction (PCR). The amplified product was entered into pTZ57R cloning vector by T/A cloning method and transformation was performed by heat shocking of the E. coli XL1-blue competent cells. Following plasmid preparation, the cloned gene was digested out and ligated into the expression vector pET-22b(+). Electrophoresis of PCR product showed a 789 bp band. Recombinant plasmid (rpTZ57R) was then constructed. This plasmid was double digested with XhoI and EcoRI resulting in 800 bp and 2900 bp bands. The obtained insert was ligated into pET-22b(+) vector and its orientation was confirmed with XhoI and BamHI restriction enzymes. In conclusion, in the present study the recombinant expression vector containing xylitol dehydrogenase gene has been constructed and can be used for the production of this enzyme in high quantities. PMID:22110522

  20. Cloning and expression of special F protein from human liver

    PubMed Central

    Liu, Shu-Ye; Yu, Xin-Da; Song, Chun-Juan; Lu, Wei; Zhang, Jian-Dong; Shi, Xin-Rong; Duan, Ying; Zhang, Ju

    2007-01-01

    AIM: To clone human liver special F protein and to express it in a prokaryotic system. METHODS: Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse primer. Following this, cDNA of the F protein was ligated into the clone vector pUCm-T. The segment of F protein’s cDNA was subcloned into the expression vector pET-15b and transformed into E. coli BL21 (DE3) pLyss. Isopropy-β-D-thiogalactoside (IPTG) was then used to induce expression of the target protein. RESULTS: The cDNA clone of human liver special F protein (1134bp) was successfully produced, with the cDNA sequence being published in Gene-bank: DQ188836. We confirmed the expression of F protein by Western blot with a molecular weight of 43 kDa. The expressed protein accounted for 40% of the total protein extracted. CONCLUSION: F protein expresses cDNA clone in a prokaryotic system, which offers a relatively simple way of producing sufficient quantities of F protein and contributes to understanding the principal biological functions of this protein. PMID:17465469

  1. Molecular cloning of protein-based polymers.

    PubMed

    Mi, Lixin

    2006-07-01

    Protein-based biopolymers have become a promising class of materials for both biomedical and pharmaceutical applications, as they have well-defined molecular weights, monomer compositions, as well as tunable chemical, biological, and mechanical properties. Using standard molecular biology tools, it is possible to design and construct genes encoding artificial proteins or protein-based polymers containing multiple repeats of amino acid sequences. This article reviews some of the traditional methods used for constructing DNA duplexes encoding these repeat-containing genes, including monomer generation, concatemerization, iterative oligomerization, and seamless cloning. A facile and versatile method, called modules of degenerate codons (MDC), which uses PCR and codon degeneracy to overcome some of the disadvantages of traditional methods, is introduced. Re-engineering of the random coil spacer domain of a bioactive protein, WPT2-3R, is used to demonstrate the utility of the MDC method. MDC re-constructed coding sequences facilitate further manipulations, such as insertion, deletion, and swapping of various sequence modules. A summary of some promising emerging techniques for synthesizing repetitive sequence-containing artificial proteins is also provided. PMID:16827576

  2. Reversibility of continuous-variable quantum cloning

    SciTech Connect

    Filip, Radim; Marek, Petr; Fiurasek, Jaromir

    2004-01-01

    We analyze a reversibility of optimal Gaussian 1{yields}2 quantum cloning of a coherent state using only local operations on the clones and classical communication between them and propose a feasible experimental test of this feature. Performing Bell-type homodyne measurement on one clone and anticlone, an arbitrary unknown input state (not only a coherent state) can be restored in the other clone by applying appropriate local unitary displacement operation. We generalize this concept to a partial reversal of the cloning using only local operations and classical communication (LOCC) and we show that this procedure converts the symmetric cloner to an asymmetric cloner. Further, we discuss a distributed LOCC reversal in optimal 1{yields}M Gaussian cloning of coherent states which transforms it to optimal 1{yields}M{sup '} cloning for M{sup '}cloning as a possible eavesdropping attack on quantum communication link, the reversibility can be utilized to improve the security of the link even after the attack.

  3. Cloning of endangered mammalian species: any progress?

    PubMed

    Loi, Pasqualino; Galli, Cesare; Ptak, Grazyna

    2007-05-01

    Attempts through somatic cell nuclear transfer to expand wild populations that have shrunk to critical numbers is a logical extension of the successful cloning of mammals. However, although the first mammal was cloned 10 years ago, nuclear reprogramming remains phenomenological, with abnormal gene expression and epigenetic deregulation being associated with the cloning process. In addition, although cloning of wild animals using host oocytes from different species has been successful, little is known about the implication of partial or total mitochondrial DNA heteroplasmy in cloned embryos, fetuses and offspring. Finally, there is a need for suitable foster mothers for inter-intra specific cloned embryos. Considering these issues, the limited success achieved in cloning endangered animals is not surprising. However, optimism comes from the rapid gain in the understanding of the molecular clues underlying nuclear reprogramming. If it is possible to achieve a controlled reversal of the differentiated state of a cell then it is probable that other issues that impair the cloning of endangered animals, such as the inter-intra species oocyte or womb donor, will be overcome in the medium term.

  4. Impact of cloning on cattle breeding systems.

    PubMed

    McClintock, A E

    1998-01-01

    The concept of clone-family testing is compared with existing progeny testing systems. The critical factors that will decide how cloning is utilized are the potential size of cloned families, and the cost per embryo (or per calf born). If family sizes of 100,000 become routinely achievable (cheaply), then clone testing becomes viable. In rough figures, cloned embryos costing $30 with a 50% calving rate would be attractive to farmers and would be cheap enough that farmers would buy more (crossbred) embryos in order to breed further replacement cows. At $300 per embryo, farmers would be more inclined to buy a number of cloned pure-bred female embryos and then to use conventional artificial insemination to breed further replacements from these superior cows. At $3000 per embryo, farmers would probably only be interested in very small numbers of cloned animals, most of which would be males. The relative importance of adult versus fetal cloning is discussed. The need for gene banks to preserve genetic variation is emphasized; both gametes and somatic tissue cultures should be considered.

  5. The ethics of human reproductive cloning.

    PubMed

    Strong, Carson

    2005-03-01

    This article addresses the question of whether human reproductive cloning could be ethically justifiable in at least some cases involving infertile couples who would choose cloning as a way to have a genetically related child. At present, the risk of congenital anomalies constitutes a compelling argument against human reproductive cloning. The article explores whether reproductive cloning could be ethically justifiable if, at some future time, cloning becomes possible without an elevated risk of anomalies. It is argued that freedom to use cloning is a form of procreative freedom and, as such, deserves respect. All of the objections that have been raised against human reproductive cloning fall under three main categories: those that appeal to the interests of the child, those based on consequences for society, and those arising from teleological views. Objections that appeal to the child's interests are, in turn, of two main kinds: consequentialist and deontological. All of these types of objections are examined, and it is found that each involves serious problems that prevent it from being a reasonable objection in the context of the infertility cases considered. It is concluded that human reproductive cloning would be ethically justifiable in at least some cases involving infertile couples, provided that it could be performed without an elevated risk of anomalies.

  6. Cloning pigs: advances and applications.

    PubMed

    Polejaeva, I A

    2001-01-01

    Although mouse embryonic stem cells have been used widely for over a decade as an important tool for introducing precise genetic modification into the genome, demonstrating the great value of this technology in a range of biomedical applications, similar technology does not exist for domestic animals. However, the development of somatic cell nuclear transfer has bypassed the need for embryonic stem cells from livestock. The production of offspring from differentiated cell nuclei provides information and opportunities in a number of areas including cellular differentiation, early development and ageing. However, the primary significance of cloning is probably in the opportunities that this technology brings to genetic manipulation. Potential applications of gene targeting in livestock species are described with particular emphasis on the generation of pigs that can be used for xenotransplantation, and the production of improved models for human physiology and disease. The development of techniques for somatic cell nuclear transfer in pigs and the challenges associated with this technology are also reviewed.

  7. Future and applications of cloning.

    PubMed

    Trounson, Alan O

    2006-01-01

    The birth of viable offspring from somatic cell nuclear transfer (SCNT) in mammals caused a major re-examination of the understanding of the commitment of cells to specific tissue lineages during differentiation. The questions of whether cells undergo dedifferentiation or transdifferentiation during the development of offspring and how these changes are controlled is a source of ongoing debate that is yet to be resolved. Irrespective of the outcome of this debate, it is clear that cloning using SCNT has a place and purpose in the future of research and animal breeding. The future uses of SCNT could include the production of transgenic mice, the production of transgenic livestock and assisting with the re-establishment of endangered species. Human medicine also would benefit from future use of SCNT because it would allow the production of patient-specific embryonic stem cells.

  8. Meat and milk compositions of bovine clones.

    PubMed

    Tian, X Cindy; Kubota, Chikara; Sakashita, Kunihito; Izaike, Yoshiaki; Okano, Ryoichi; Tabara, Norio; Curchoe, Carol; Jacob, Lavina; Zhang, Yuqin; Smith, Sadie; Bormann, Charles; Xu, Jie; Sato, Masumi; Andrew, Sheila; Yang, Xiangzhong

    2005-05-01

    The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones.

  9. [Human clone or a delayed twin?].

    PubMed

    Szybalski, W

    2001-01-01

    Cloning is a natural mode of asexual reproduction for many organisms, which results in nearly identical copies of cells or organisms. In animals, including humans, identical twins are an example of natural cloning. In the case of sheep, scientists succeeded to produce the "delayed" identical twin. Dolly, of a mature animal by a rather complex and inefficient procedure. However, if this procedure is perfected, it will be useful to clone beloved pets and important laboratory animals. It will be much less suited for making (cloning) "delayed twin" of mature persons because of high costs together with present experimental uncertainties. The only required regulation for human cloning is that somebody must be legally, including financially, responsible for the results of such novel reproductive technique.

  10. [Human cloning in Muslim and Arab law].

    PubMed

    Aldeeb Abu-Sahlieh, Sami A

    2009-01-01

    Cloning is a modern medical procedure that Muslim religious authorities treat en resorting to the general principles established by classical Muslim law based on the Koran and the Sunnah of Muhhamad as the messenger of God. In this regard, human beings are not capable of deciding what is or what is not lawful without resorting to divine norms. Cloning clashes with several principles. Firstly, the principle of the respect for life in relation to surpernumeraries, but Muslim authors are not in unanimous agreement on the determination of the moment at which life begins. Secondly, is the respect of progeny: cloning could only take place between a married couple. But even if these two principles are respected, cloning poses two major problems: the diversity of species expounded by the Koran and the Sunnah and a lack of interest. Which explains the quasi-unanimous opposition of Muslim writings regarding cloning.

  11. "Goodbye Dolly?" The ethics of human cloning.

    PubMed

    Harris, J

    1997-12-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity.

  12. Chorioallantoic placenta defects in cloned mice

    SciTech Connect

    Wakisaka-Saito, Noriko; Kohda, Takashi . E-mail: tkhoda.epgn@tmd.ac.jp; Inoue, Kimiko; Ogonuki, Narumi; Miki, Hiromi; Hikichi, Takafusa; Mizutani, Eiji; Wakayama, Teruhiko; Kaneko-Ishino, Tomoko; Ogura, Atsuo; Ishino, Fumitoshi

    2006-10-13

    Somatic cell nuclear transfer technology has been applied to produce live clones successfully in several mammalian species, but the success rates are very low. In mice, about half of the nuclear transfer embryos undergo implantation, but very few survive to term. We undertook detailed histological analyses of placentas from cloned mouse embryos generated from cumulus cells at 10.5 dpc of pregnancy, by which stage most clones have terminated their development. At 10.5 dpc, the extraembryonic tissues displayed several defined histological patterns, each reflecting their stage of developmental arrest. The most notable abnormality was the poor development of the spongiotrophoblast layer of diploid cells. This is in contrast to the placental hyperplasia frequently observed in somatic clones at 12.5 dpc or later stages. A variety of structural abnormalities were also observed in the embryos. Both placental and embryonic defects likely contribute to the low success rate of the mouse clones.

  13. "Goodbye Dolly?" The ethics of human cloning.

    PubMed Central

    Harris, J

    1997-01-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity. PMID:9451604

  14. Normal telomere lengths found in cloned cattle.

    PubMed

    Tian, X C; Xu, J; Yang, X

    2000-11-01

    Success of cloning using adult somatic cells has been reported in sheep, mice and cattle. The report that 'Dolly' the sheep, the first clone from an adult mammal, inherited shortened telomeres from her cell donor and that her telomeres were further shortened by the brief culture of donor cells has raised serious scientific and public concerns about the 'genetic age' and potential developmental problems of cloned animals. This observation was challenged by a recent report that showed calves cloned from fetal cells have longer telomeres than their age-matched controls. The question remains whether Dolly's short telomeres were an exception or a general fact, which would differ from the telomeres of fetal-derived clones. PMID:11062462

  15. [Human clone or a delayed twin?].

    PubMed

    Szybalski, W

    2001-01-01

    Cloning is a natural mode of asexual reproduction for many organisms, which results in nearly identical copies of cells or organisms. In animals, including humans, identical twins are an example of natural cloning. In the case of sheep, scientists succeeded to produce the "delayed" identical twin. Dolly, of a mature animal by a rather complex and inefficient procedure. However, if this procedure is perfected, it will be useful to clone beloved pets and important laboratory animals. It will be much less suited for making (cloning) "delayed twin" of mature persons because of high costs together with present experimental uncertainties. The only required regulation for human cloning is that somebody must be legally, including financially, responsible for the results of such novel reproductive technique. PMID:11684762

  16. Crops Demand-Side Irrigation Principle and Practice-The Innovation Irrigation Method to Save World's Water Supply in 21st Century

    NASA Astrophysics Data System (ADS)

    Fok, Y.

    2001-12-01

    More than 75% of the world's water suypply is used for irrigation. Currently the world's irrigation water use efficiency is less than 40%. If irrigation water use efficiency can be improved to 60% and higher, large quantity of water supply would be saved. In order to attend a higher irrigation water use efficiency in 21st century, an exammination of current irrigation principle and practice is needed. The current irrigation practice is based on the public water supply principle which uses the supply-side principle. Irrigation systems are designed by engineers, who design the irrigation systems to supply water to irrigation the farmlands at a scheduled frequency to support plant growth, they disregard the demand of water or not by the crops. This is the short-coming of the supply-side irrigation principle. Because it often over-irrigate the farmlands and frequently causes water logging and groundwater contammination. The principle of the demand-side crop irrigation is to satify the water demand of the crops when they need it and turn-off the water when their demand have been satisfied automatically. The advances in computer technology have given us the needed tools to fully accommodate the innovation and automation of the demand-side irrigation systems. The author expect to see a large amount of new hardwares on the market in 21st century. For example, the drip irrigation systems are in a position for changing into the demand-side irrigation systems readily. The emitters of the drip irrigation systems can be modified into the non-plugging and soil-misture tension sensing outlets to suit the demand-side irrigation principle to satisfy crops water demand and turn-off when the demand had been satified. Other innovative invention patents may be developed for sub-irrigation, sprinkler irrigation, border strip irrigation and furrow irrigation to solve the world's water supply shortage problems by means of the innovations in crop demand-side irrigation principle and

  17. Cas9-Assisted Targeting of CHromosome segments CATCH enables one-step targeted cloning of large gene clusters.

    PubMed

    Jiang, Wenjun; Zhao, Xuejin; Gabrieli, Tslil; Lou, Chunbo; Ebenstein, Yuval; Zhu, Ting F

    2015-09-01

    The cloning of long DNA segments, especially those containing large gene clusters, is of particular importance to synthetic and chemical biology efforts for engineering organisms. While cloning has been a defining tool in molecular biology, the cloning of long genome segments has been challenging. Here we describe a technique that allows the targeted cloning of near-arbitrary, long bacterial genomic sequences of up to 100 kb to be accomplished in a single step. The target genome segment is excised from bacterial chromosomes in vitro by the RNA-guided Cas9 nuclease at two designated loci, and ligated to the cloning vector by Gibson assembly. This technique can be an effective molecular tool for the targeted cloning of large gene clusters that are often expensive to synthesize by gene synthesis or difficult to obtain directly by traditional PCR and restriction-enzyme-based methods.

  18. Subtractive cloning: new genes for studying inflammatory disorders.

    PubMed

    Rebrikov, Denis; Desai, Sejal; Kogan, Yakov N; Thornton, Angela M; Diatchenko, Luda

    2002-12-01

    Understanding of the biology of interaction between pathogens and host is the central question in studying inflammatory disorders. Subtractive DNA cloning is one of the most efficient and comprehensive methods available for identifying eukaryotic genes regulated under specific physiological conditions, including inflammation and host response. Here we explore the utility of subtractive DNA cloning and describe suppression subtractive hybridization (SSH), a polymerase chain reaction (PCR)-based DNA subtraction method that has been developed and evolved in our labs over several years. The SSH method possesses a number of advantages as compared to other subtractive cloning techniques, making it one of the most adventitious methods for cloning differentially expressed genes. Besides isolation of differentially expressed eukaryotic mRNAs, subtractive DNA cloning can be used to identify genes that are differentially expressed between diverse bacterial species. These genes can be of great interest, as some may encode strain-specific traits such as drug resistance, or bacterial surface proteins involved in determining the virulence of a particular strain. Other genes may be useful as markers for epidemiological or evolutionary studies. To demonstrate the potential of the SSH technique, we describe here the comprehensive characterization of 2 SSH subtracted libraries constructed in our laboratories. One library was created using eukaryotic cDNA subtraction and is specific for mRNAs up-regulated in CD25 positive cells from mouse lymph nodes as compared to CD25 negative cells. The second subtracted library is specific for a methicillin-resistant Staphylococcus aureus bacterial strain, but not in a methicillin-sensitive strain. The bacterial genomes of these 2 strains have been completely sequenced and this second library provides an excellent reference for testing the ability of SSH to recover all strain-specific gene content. The analysis of these 2 subtracted libraries

  19. Using Educational Innovations to Meet Student Needs

    ERIC Educational Resources Information Center

    McSweeney, Jean, Ed.

    1977-01-01

    Summarizes 18 papers given at the Fourth Biennial Conference on Chemical Education that dealt with innovative instructional methods or theories, including instructional objectives, audiotape-slide programs, and criterion-referenced evaluation. (MLH)

  20. Measuring the style of innovative thinking among engineering students

    NASA Astrophysics Data System (ADS)

    Passig, David; Cohen, Lizi

    2014-01-01

    Background: Many tools have been developed to measure the ability of workers to innovate. However, all of them are based on self-reporting questionnaires, which raises questions about their validity Purpose: The aim was to develop and validate a tool, called Ideas Generation Implementation (IGI), to objectively measure the style and potential of engineering students in generating innovative technological ideas. The cognitive framework of IGI is based on the Architectural Innovation Model (AIM). Tool description: The IGI tool was designed to measure the level of innovation in generating technological ideas and their potential to be implemented. These variables rely on the definition of innovation as 'creativity, implemented in a high degree of success'. The levels of innovative thinking are based on the AIM and consist of four levels: incremental innovation, modular innovation, architectural innovation and radical innovation. Sample: Sixty experts in technological innovation developed the tool. We checked its face validity and calculated its reliability in a pilot study (kappa = 0.73). Then, 145 undergraduate students were sampled at random from the seven Israeli universities offering engineering programs and asked to complete the questionnaire. Design and methods: We examined the construct validity of the tool by conducting a variance analysis and measuring the correlations between the innovator's style of each student, as suggested by the AIM, and the three subscale factors of creative styles (efficient, conformist and original), as suggested by the Kirton Adaptors and Innovators (KAI) questionnaire. Results: Students with a radical innovator's style inclined more than those with an incremental innovator's style towards the three creative cognitive styles. Students with an architectural innovator's style inclined moderately, but not significantly, towards the three creative styles. Conclusions: The IGI tool objectively measures innovative thinking among students