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 lymphoblastoid cells

    SciTech Connect

    Hammerling, U.; Kosinski, S.

    1989-02-14

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

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

    PubMed

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2014-01-01

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

  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. Teaching methods: Tradition and innovation

    NASA Astrophysics Data System (ADS)

    Nikandrov, Nikolai D.

    1990-06-01

    It has always been difficult to distinguish between tradition and innovation in teaching methods, not least because of the absence of clear-cut criteria. Definitions of teaching methods are also loose rather than binding. Nevertheless a trend towards active participation by studients is noticeable and very often taken as marking `progressive' teaching. Starting from the basic relationship of method and objective, an attempt is made to further relate cognitive activity of students to specific levels of achievement which are considered as teaching objectives. It is suggested, too, that a loose notion of method can for practical purposes be replaced by a more reliable notion of teaching text whether presented orally or given in written form. Then the problem of innovation in teaching methods can be stated a bit more precisely as that of creating a good teaching text. Some suggestions of how this can be achieved are discussed.

  6. Technological Innovation and Teaching Methods

    NASA Astrophysics Data System (ADS)

    Riboli, Maria Cristina

    The wide diffusion of information technology brins generally changes in any field of society. In universities, the teaching approach must conform itself to these new needs adding the information technology tools into the learning process. A multi-disciplinary technical team of Florence University was set up a project involving technical staff developer, educational technologist and web developer and has pointed out issues concerning hardware, software, technical methods and knowledge required by these innovations.

  7. A modified version of the digestion-ligation cloning method for more efficient molecular cloning.

    PubMed

    Gao, Song; Li, Yanling; Zhang, Jiannan; Chen, Hongman; Ren, Daming; Zhang, Lijun; An, Yingfeng

    2014-05-15

    Here we describe a modified version of the digestion-ligation approach for efficient molecular cloning. In comparison with the original method, the modified method has the additional steps of gel purification and a second ligation after the first ligation of the linearized vector and DNA insert. During this process, the efficiency and reproducibility could be significantly improved for both stick-end cloning and blunt-end cloning. As an improvement of the very important molecular cloning technique, this method may find a wide range of applications in bioscience and biotechnology.

  8. 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.

  9. 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

  10. Cloning

    MedlinePlus

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

  11. Development of an in vitro cloning method for Cowdria ruminantium.

    PubMed Central

    Perez, J M; Martinez, D; Debus, A; Sheikboudou, C; Bensaid, A

    1997-01-01

    Cowdria ruminantium is a tick-borne rickettsia which causes severe disease in ruminants. All studies with C. ruminantium reported so far were carried out with stocks consisting of infective blood collected from reacting animals or from the same stocks propagated in vitro. Cloned isolates are needed to conduct studies on immune response of the host, on genetic diversity of the parasite, and on mechanisms of attenuation and the development of vaccines. A method of cloning based on the particular chlamydia life cycle of Cowdria was developed. Instead of cloning extracellular elementary bodies, it appeared more convenient to clone endothelial cells infected by one morula resulting from the infection of the cell by one elementary body of Cowdria. Two hundred and sixteen clones were obtained by limiting dilution of infected cells. The method was experimentally validated by comparing randomly amplified polymorphic DNA fingerprints from individual clones obtained from endothelial cell cultures coinfected with two different stocks of C. ruminantium. PMID:9302217

  12. Innovative methods speed soil cleanup

    SciTech Connect

    Alexander, T.G.

    1994-05-01

    Innovative technolgies for soil remediation are discussed, as well as hurdles towards their use. Technologies include soil washing, vapor extraction, bioremediation, chemical treatment, and other forms of themal treatment. Use at Superfund sites is described.

  13. To Clone or Not To Clone: Method Analysis for Retrieving Consensus Sequences In Ancient DNA Samples

    PubMed Central

    Winters, Misa; Barta, Jodi Lynn; Monroe, Cara; Kemp, Brian M.

    2011-01-01

    The challenges associated with the retrieval and authentication of ancient DNA (aDNA) evidence are principally due to post-mortem damage which makes ancient samples particularly prone to contamination from “modern” DNA sources. The necessity for authentication of results has led many aDNA researchers to adopt methods considered to be “gold standards” in the field, including cloning aDNA amplicons as opposed to directly sequencing them. However, no standardized protocol has emerged regarding the necessary number of clones to sequence, how a consensus sequence is most appropriately derived, or how results should be reported in the literature. In addition, there has been no systematic demonstration of the degree to which direct sequences are affected by damage or whether direct sequencing would provide disparate results from a consensus of clones. To address this issue, a comparative study was designed to examine both cloned and direct sequences amplified from ∼3,500 year-old ancient northern fur seal DNA extracts. Majority rules and the Consensus Confidence Program were used to generate consensus sequences for each individual from the cloned sequences, which exhibited damage at 31 of 139 base pairs across all clones. In no instance did the consensus of clones differ from the direct sequence. This study demonstrates that, when appropriate, cloning need not be the default method, but instead, should be used as a measure of authentication on a case-by-case basis, especially when this practice adds time and cost to studies where it may be superfluous. PMID:21738625

  14. USER-derived cloning methods and their primer design.

    PubMed

    Salomonsen, Bo; Mortensen, Uffe H; Halkier, Barbara A

    2014-01-01

    Uracil excision-based cloning through USER™ (Uracil-Specific Excision Reagent) is an efficient ligase-free cloning technique that comprises USER cloning, USER fusion, and USER cassette-free (UCF) USER fusion. These USER-derived cloning techniques enable seamless assembly of multiple DNA fragments in one construct. Though governed by a few simple rules primer design for USER-based fusion of PCR fragments can prove time-consuming for inexperienced users. The Primer Help for USER (PHUSER) software is an easy-to-use primer design tool for USER-based methods. In this chapter, we present a PHUSER software protocol for designing primers for USER-derived cloning techniques.

  15. Cloning

    MedlinePlus

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

  16. A method of grid element cloning

    NASA Astrophysics Data System (ADS)

    Bednyakov, I. V.; Dolbilov, A. G.; Ivanov, Yu. P.

    2010-11-01

    A simple, reliable, and fast method of putting a new computational facilities into operation in a grid environment is described. This method was successfully tested as part of the grid segment at Dzhelepov Laboratory of Nuclear Problems, Joint Institute for Nuclear Research (JINR), for work at the ATLAS collaboration.

  17. 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)

  18. 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.

  19. Innovative methods of popularizing technical education

    NASA Astrophysics Data System (ADS)

    Shkitsa, L. Y.; Panchuk, V. G.; Kornuta, V. A.

    2017-05-01

    There have been analyzed reasons of the loss of technical education’s popularity. Also, the analysis of known educational and production methods, oriented at the innovative model of development of society, was performed. It is stated that the acquisition of 21st century’s skills as a result of competition of technical education are natural for the DIY ideology, which was realized in the institutions like Fab Lab. The new educational strategy, based on project-based learning, is proposed to be implemented as a special laboratory with equipment, which would be a center of innovative development for students at the Technical University. Moreover, the list of projects planned for implementation, that includes not only projects, specific to a particular university, but also projects, demanded by society as a whole, is specified. It is worth to implement trendy projects in the laboratory, such as toy-like, ecological projects; projects of the energy dependence decrease or the energy efficiency increase, modern digital or innovative projects etc. The student should gain knowledge, skills and, possibly, equipment that are available for immediate usage on the labor market or for the realization of his own projects or the community’s projects in everyday life after the realization of the particular project at the laboratory

  20. 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. © The Author(s) 2015.

  1. 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.

  2. Recent advances in universal TA cloning methods for use in function studies.

    PubMed

    Yao, Shuo; Hart, Darren J; An, Yingfeng

    2016-08-29

    As one of the simplest and most efficient cloning methods, T-vector-based TA cloning has been widely used for cloning of single genes and construction of DNA libraries. This approach is especially suitable for high-throughput cloning of diverse DNA fragments since inserts can be cloned without knowledge of their sequence; it is therefore an ideal tool for high-throughput analysis of protein structure and function. Although most of the currently available T-vectors can only be used for cloning purposes, some novel variants with improved functions have be developed. This review focuses on recent developments of universal TA cloning methods and T-vectors constructed for function studies.

  3. 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.

  4. Methods for Studying Innovation Development in the Minnesota Innovation Research Program.

    ERIC Educational Resources Information Center

    Van de Ven, Andrew H.; Poole, Marshall Scott

    1990-01-01

    Focuses on the methods being used to examine processes of innovation development that pertain to the selection of cases and concepts, observing change, coding and analyzing event data to identify process patterns, and developing theories to explain observed innovation processes. (42 references) (MLF)

  5. An efficient method to prepare PCR cloning vectors.

    PubMed

    Hong, Soon Gyu; Choi, Ji Young; Pryor, Barry M; Lee, Hong Kum

    2009-09-01

    An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation of adapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by further digestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purification kits. Using this procedure, PCR cloning vectors with high ligation efficiencies and low blue or false-positive colonies were obtained.

  6. 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.

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

    PubMed

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

    2016-07-08

    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. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  8. 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

  9. 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...

  10. 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...

  11. A general method for saturation mutagenesis of cloned DNA fragments.

    PubMed

    Myers, R M; Lerman, L S; Maniatis, T

    1985-07-19

    A new procedure for generating and isolating random single-base substitutions in cloned DNA fragments is presented. The mutations are generated by treatment of single-stranded DNA with various chemicals, followed by the synthesis of the complementary strand with reverse transcriptase. Misincorporation frequently occurs when the enzyme encounters a damaged base in the mutagenized template DNA. The resulting duplex DNA fragments containing random single-base substitutions are cloned, amplified as a population, and isolated from wild-type DNA by preparative denaturing gradient gel electrophoresis. The physical separation of mutant DNA fragments makes it possible to isolate and characterize large numbers of site-directed single-base substitutions in the absence of a phenotypic selection. This procedure should be generally applicable to the fine-structure genetic analysis of regulatory and protein-coding sequences.

  12. 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.

  13. 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

  14. IRDL cloning: a one-tube, zero-background, easy-to-use, directional cloning method improves throughput in recombinant DNA preparation.

    PubMed

    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.

  15. Innovation

    EPA Pesticide Factsheets

    EPA frames innovation as critical to the protection of human health and the environment through initiatives such as sustainable practices, innovative research, prize competitions, innovation awards, partnerships, and community activities.

  16. Innovative Methods for High Resolution Imaging

    DTIC Science & Technology

    2012-08-02

    Space Telescope . A data volume of 177 × 193 × 100 was successfully processed. 5 Technology Transfer 5.1 Mitigating Pulsed Laser Attack on CCD-based...01 2007) 2012/07/16 12:20:07 3 Qiang Zhang, Han Wang, Robert J. Plemmons, V. Paul Pauca. Tensor Methods for Hyperspectral Data Analysis: A Space ...ISBN: 978-0-387- 73002-8), 2009. 2. P. Zhang, H. Wang, R.J. Plemmons, and V.P. Pauca. “Tensor Methods for Hyperspec- tral Analysis of Space Objects, A

  17. 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.

  18. Innovative methods for inorganic sample preparation

    SciTech Connect

    Essling, A.M.; Huff, E.A.; Graczyk, D.G.

    1992-04-01

    Procedures and guidelines are given for the dissolution of a variety of selected materials using fusion, microwave, and Parr bomb techniques. These materials include germanium glass, corium-concrete mixtures, and zeolites. Emphasis is placed on sample-preparation approaches that produce a single master solution suitable for complete multielement characterization of the sample. In addition, data are presented on the soil microwave digestion method approved by the Environmental Protection Agency (EPA). Advantages and disadvantages of each sample-preparation technique are summarized.

  19. A simple method for generating full length cDNA from low abundance partial genomic clones

    PubMed Central

    Wang, Yongxin; Fugaro, Joseph M; Siddiq, Fauzia; Goparaju, Chandra Mouli V; Lonardo, Fulvio; Wali, Anil; Lechner, John F; Pass, Harvey I

    2000-01-01

    Background PCR amplification of target molecules involves sequence specific primers that flank the region to be amplified. While this technique is generally routine, its applicability may not be sufficient to generate a desired target molecule from two separate regions involving intron /exon boundaries. For these situations, the generation of full-length complementary DNAs from two partial genomic clones becomes necessary for the family of low abundance genes. Results The first approach we used for the isolation of full-length cDNA from two known genomic clones of Hox genes was based on fusion PCR. Here we describe a simple and efficient method of amplification for homeobox D13 (HOXD13) full length cDNA from two partial genomic clones. Specific 5' and 3' untranslated region (UTR) primer pairs and website program (primer3_www.cgv0.2) were key steps involved in this process. Conclusions We have devised a simple, rapid and easy method for generating cDNA clone from genomic sequences. The full length HOXD13 clone (1.1 kb) generated with this technique was confirmed by sequence analysis. This simple approach can be utilized to generate full-length cDNA clones from available partial genomic sequences. PMID:11114844

  20. A rapid cloning method employing orthogonal end protection.

    PubMed

    Jakobi, Arjen J; Huizinga, Eric G

    2012-01-01

    We describe a novel in vitro cloning strategy that combines standard tools in molecular biology with a basic protecting group concept to create a versatile framework for the rapid and seamless assembly of modular DNA building blocks into functional open reading frames. Analogous to chemical synthesis strategies, our assembly design yields idempotent composite synthons amenable to iterative and recursive split-and-pool reaction cycles. As an example, we illustrate the simplicity, versatility and efficiency of the approach by constructing an open reading frame composed of tandem arrays of a human fibronectin type III (FNIII) domain and the von Willebrand Factor A2 domain (VWFA2), as well as chimeric (FNIII)(n)-VWFA2-(FNIII)(n) constructs. Although we primarily designed this strategy to accelerate assembly of repetitive constructs for single-molecule force spectroscopy, we anticipate that this approach is equally applicable to the reconstitution and modification of complex modular sequences including structural and functional analysis of multi-domain proteins, synthetic biology or the modular construction of episomal vectors.

  1. Cloning cattle.

    PubMed

    Oback, B; Wells, D N

    2003-01-01

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

  2. Carisolv- an innovative method of caries removal.

    PubMed

    Kathuria, Vartika; Ankola, Anil V; Hebbal, Mamata; Mocherla, Monica

    2013-12-01

    The use of minimal invasive dentistry and attention to patient comfort is of utmost importance especially for the school children and anxious and uncooperative patients. This demanded the newer patient friendly technique in dentistry amongst which is the Chemo mechanical caries removal (CMCR). CMCR involves the selective caries removal of carious dentine. As only the carious dentine is removed the painful removal of sound dentine is avoided and the anxiety due to the vibration of the handpiece is also reduced thus it appears to be more comfortable to the patient. CMCR although more prolonged procedure, is effective in caries removal. Thus CMCR is an effective alternate method for caries removal; it is more conservative and appears to be more comfortable for the patients.

  3. Innovative method and equipment for personalized ventilation.

    PubMed

    Kalmár, F

    2015-06-01

    At the University of Debrecen, a new method and equipment for personalized ventilation has been developed. This equipment makes it possible to change the airflow direction during operation with a time frequency chosen by the user. The developed office desk with integrated air ducts and control system permits ventilation with 100% outdoor air, 100% recirculated air, or a mix of outdoor and recirculated air in a relative proportion set by the user. It was shown that better comfort can be assured in hot environments if the fresh airflow direction is variable. Analyzing the time step of airflow direction changing, it was found that women prefer smaller time steps and their votes related to thermal comfort sensation are higher than men's votes.

  4. Implications of cloning for breed improvement strategies: are traditional methods of animal improvement obsolete?

    PubMed

    van Vleck, L D

    1999-01-01

    Can the optimum animal be defined? Will that definition change over time, by location, by market demand? First, assume what may be impossible, that the perfect animal can be defined or that only a limited number of definitions of "perfect" are needed. Then, can the "perfect" animal to match a definition be found? Suppose such an animal is found. Then the question to be answered before trying to clone as a method of genetic improvement becomes "Is the animal perfect because of phenotype or genotype?" In other words, the P = G + E problem exists, which requires traditional methods of genetic evaluation and testing to determine whether genotype (G) or random environmental (E) effects or a combination leads to the apparent perfection in the phenotype (P). For most traits, additive genetic variance accounts for 10 to 50% of total variance, a fraction denoted as heritability. With a simple model, the best prediction of genotypic value is to reduce the apparent phenotypic superiority by multiplying by heritability. Cloning the "perfect" animal also could capture optimum dominance and epistatic genetic effects that are otherwise difficult to select for. For some traits, maternal effects are important. In that case, clones as breeding animals must be "perfect" for both direct and maternal genotypes, or alternatively terminal and maternal clone lines would need to be developed. The use of clones to increase uniformity can be only partially successful. If heritability is 25%, then the standard deviation among clones would be 87% of that of uncloned animals. Only if heritability is 100% will clone mates have complete uniformity. Fixing the genotype could increase susceptibility to failure if environment changes or if the cloned genotype is susceptible to a new disease or if economic conditions change. Cloning, at best, is another tool for animal improvement that joins the list of previous biotechnological inventions, some of which have become cost-effective, such as artificial

  5. Innovative Methods and Applications in Mucoadhesion Research.

    PubMed

    Mackie, Alan R; Goycoolea, Francisco M; Menchicchi, Bianca; Caramella, Carla Marcella; Saporito, Francesca; Lee, Seunghwan; Stephansen, Karen; Chronakis, Ioannis S; Hiorth, Marianne; Adamczak, Malgorzata; Waldner, Max; Nielsen, Hanne Mørck; Marcelloni, Luciano

    2017-08-01

    The present review is aimed at elucidating relatively new aspects of mucoadhesion/mucus interaction and related phenomena that emerged from a Mucoadhesion workshop held in Munster on 2-3 September 2015 as a satellite event of the ICCC 13th-EUCHIS 12th. After a brief outline of the new issues, the focus is on mucus description, purification, and mucus/mucin characterization, all steps that are pivotal to the understanding of mucus related phenomena and the choice of the correct mucosal model for in vitro and ex vivo experiments, alternative bio/mucomimetic materials are also presented. Then a selection of preparative techniques and testing methods are described (at molecular as well as micro and macroscale) that may support the pharmaceutical development of mucus interactive systems and assist formulators in the scale-up and industrialization steps. Recent applications of mucoadhesive systems (including medical devices) intended for different routes of administration (oral, gastrointestinal, vaginal, nasal, ocular, and intravesical) and for the treatment of difficult to treat pathologies or the alleviation of symptoms are described. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. 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…

  7. A simple method for developing an infectious cDNA clone of Japanese encephalitis virus.

    PubMed

    Zheng, Hao; Zheng, Xuchen; Tong, Wu; Liu, Fei; Liang, Chao; Wang, Tao; Gao, Fei; Li, Liwei; Shan, Tongling; Li, Guoxin; Tong, Guangzhi

    2017-02-01

    Flavivirus cDNA clones frequently demonstrate genetic instability in transformed bacteria, which hampers the construction and manipulation of cDNAs for infectious flaviviruses. In this study, we developed a stable, full-length cDNA clone, pJEHEN, of a GI JEV strain HEN0701 using a medium-copy-number pBR322 vector and propagating cDNA clones at room temperature. The virus vJEHEN recovered from the infectious clone was indistinguishable from the parent virus HEN0701 with respect to plaque morphology, growth kinetics, and virulence characteristics. A T-to-A silent mutation of nucleotide 24 of the NS2a gene was introduced into the infectious cDNA clone to eliminate frameshifting. The rescued mutant virus vJETA did not express NS1' in infected cells and showed reduced growth and neurovirulence in mice. This convenient method for the construction and manipulation of infectious JEV cDNA clones may be of use in further studies to improve our understanding of the molecular mechanisms responsible for JEV replication and pathogenesis.

  8. Establishment of primary bovine intestinal epithelial cell culture and clone method.

    PubMed

    Zhan, Kang; Lin, Miao; Liu, Ming-Mei; Sui, Yang-Nan; Zhao, Guo-Qi

    2017-01-01

    The aim of this study was to establish bovine intestinal epithelial cell (BIEC) line and provide a novel clone cell method. Although various strategies of bovine cell culture and clone techniques have been reported, these methods remain not established. Here, we culture successfully primary BIECs and establish a novel clone cell method. Our result showed that BIECs could be successfully cultured and passaged about generation 5. These cellular aggregates and clusters were adherent loosely at day 2 of culture. Cell aggregates and clusters start to proliferate after approximately 4 d. The BIECs showed positive reaction against cytokeratin 18, E-cadherin, and characteristics of epithelial-like morphology. In addition, the fatty acid-binding proteins (FABPs), villin, and intestinal peptidase (IP) band were positive in BIECs. Our results suggest that the establishment of culturing and clone BIEC methods will apply to isolate and clone other primary cells. These BIECs could therefore contribute to the study of bovine intestinal nutrient absorption and regulation, immune regulation, and the pathogenesis of the bovine intestinal disease, which will provide intestinal cell model in vitro.

  9. Overview of post Cohen-Boyer methods for single segment cloning and for multisegment DNA assembly.

    PubMed

    Sands, Bryan; Brent, Roger

    2016-01-01

    In 1973, Cohen and coworkers published a foundational paper describing the cloning of DNA fragments into plasmid vectors. In it, they used DNA segments made by digestion with restriction enzymes and joined these in vitro with DNA ligase. These methods established working recombinant DNA technology and enabled the immediate start of the biotechnology industry. Since then, "classical" recombinant DNA technology using restriction enzymes and DNA ligase has matured. At the same time, researchers have developed numerous ways to generate large, complex, multisegment DNA constructions that offer advantages over classical techniques. Here, we provide an overview of "post-Cohen-Boyer" techniques used for cloning single segments into vectors (T/A, Topo cloning, Gateway and Recombineering) and for multisegment DNA assembly (Biobricks, Golden Gate, Gibson, Yeast homologous recombination in vivo, and Ligase Cycling Reaction). We compare and contrast these methods and also discuss issues that researchers should consider before choosing a particular multisegment DNA assembly method.

  10. Overview of post Cohen-Boyer methods for single segment cloning and for multisegment DNA assembly

    PubMed Central

    Sands, Bryan; Brent, Roger

    2016-01-01

    In 1973, Cohen and coworkers published a foundational paper describing the cloning of DNA fragments into plasmid vectors. In it, they used DNA segments made by digestion with restriction enzymes and joined these in vitro with DNA ligase. These methods established working recombinant DNA technology and enabled the immediate start of the biotechnology industry. Since then, “classical” recombinant DNA technology using restriction enzymes and DNA ligase has matured. At the same time, researchers have developed numerous ways to generate large, complex, multisegment DNA constructions that offer advantages over classical techniques. Here, we provide an overview of “post-Cohen-Boyer” techniques used for cloning single segments into vectors (T/A, Topo cloning, Gateway and Recombineering) and for multisegment DNA assembly (Biobricks, Golden Gate, Gibson, Yeast homologous recombination in vivo, and Ligase Cycling Reaction). We compare and contrast these methods and also discuss issues that researchers should consider before choosing a particular multisegment DNA assembly method. PMID:27152131

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

    PubMed

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2012-04-01

    We describe a novel cloning method termed SLiCE (Seamless Ligation Cloning Extract) that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (≥15 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from Bacteria Artificial Chromosomes (BACs) or other sources. SLiCE is highly cost effective as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. In addition, the cloning efficiencies and capabilities of these strains can be greatly improved by simple genetic modifications. As an example, we modified the DH10B Escherichia coli strain to express an optimized λ prophage Red recombination system. This strain, termed PPY, facilitates SLiCE with very high efficiencies and demonstrates the versatility of the method.

  12. A One Pot, One Step, Precision Cloning Method with High Throughput Capability

    PubMed Central

    Engler, Carola; Kandzia, Romy; Marillonnet, Sylvestre

    2008-01-01

    Current cloning technologies based on site-specific recombination are efficient, simple to use, and flexible, but have the drawback of leaving recombination site sequences in the final construct, adding an extra 8 to 13 amino acids to the expressed protein. We have devised a simple and rapid subcloning strategy to transfer any DNA fragment of interest from an entry clone into an expression vector, without this shortcoming. The strategy is based on the use of type IIs restriction enzymes, which cut outside of their recognition sequence. With proper design of the cleavage sites, two fragments cut by type IIs restriction enzymes can be ligated into a product lacking the original restriction site. Based on this property, a cloning strategy called ‘Golden Gate’ cloning was devised that allows to obtain in one tube and one step close to one hundred percent correct recombinant plasmids after just a 5 minute restriction-ligation. This method is therefore as efficient as currently used recombination-based cloning technologies but yields recombinant plasmids that do not contain unwanted sequences in the final construct, thus providing precision for this fundamental process of genetic manipulation. PMID:18985154

  13. A one pot, one step, precision cloning method with high throughput capability.

    PubMed

    Engler, Carola; Kandzia, Romy; Marillonnet, Sylvestre

    2008-01-01

    Current cloning technologies based on site-specific recombination are efficient, simple to use, and flexible, but have the drawback of leaving recombination site sequences in the final construct, adding an extra 8 to 13 amino acids to the expressed protein. We have devised a simple and rapid subcloning strategy to transfer any DNA fragment of interest from an entry clone into an expression vector, without this shortcoming. The strategy is based on the use of type IIs restriction enzymes, which cut outside of their recognition sequence. With proper design of the cleavage sites, two fragments cut by type IIs restriction enzymes can be ligated into a product lacking the original restriction site. Based on this property, a cloning strategy called 'Golden Gate' cloning was devised that allows to obtain in one tube and one step close to one hundred percent correct recombinant plasmids after just a 5 minute restriction-ligation. This method is therefore as efficient as currently used recombination-based cloning technologies but yields recombinant plasmids that do not contain unwanted sequences in the final construct, thus providing precision for this fundamental process of genetic manipulation.

  14. Review of enucleation methods and procedures used in animal cloning: state of the art.

    PubMed

    Li, Guang-Peng; White, Kenneth L; Bunch, Thomas D

    2004-01-01

    Enucleation of a recipient oocyte is a crucially important process for nuclear transfer efficiency. Several procedures have been developed and used in the production of nuclear transfer embryos. Although the use of excitable fluorochromes and ultraviolet (UV) light are commonly used for complete enucleation, they also pose the risk of damaging the maternal cytoplast. Telophase and chemically assisted enucleation have also been used for cloning, but the quality and quantity of the recipient cytoplasm varies with the procedure used. This paper reviews various methods used for enucleation, and discusses their benefits and limitations with respect to cloning efficiency.

  15. 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.

  16. 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

  17. A new method to customize protein expression vectors for fast, efficient and background free parallel cloning.

    PubMed

    Scholz, Judith; Besir, Hüseyin; Strasser, Claudia; Suppmann, Sabine

    2013-02-14

    Expression and purification of correctly folded proteins typically require screening of different parameters such as protein variants, solubility enhancing tags or expression hosts. Parallel vector series that cover all variations are available, but not without compromise. We have established a fast, efficient and absolutely background free cloning approach that can be applied to any selected vector. Here we describe a method to tailor selected expression vectors for parallel Sequence and Ligation Independent Cloning. SLIC cloning enables precise and sequence independent engineering and is based on joining vector and insert with 15-25 bp homologies on both DNA ends by homologous recombination. We modified expression vectors based on pET, pFastBac and pTT backbones for parallel PCR-based cloning and screening in E.coli, insect cells and HEK293E cells, respectively. We introduced the toxic ccdB gene under control of a strong constitutive promoter for counterselection of insert less vector. In contrast to DpnI treatment commonly used to reduce vector background, ccdB used in our vector series is 100% efficient in killing parental vector carrying cells and reduces vector background to zero. In addition, the 3' end of ccdB functions as a primer binding site common to all vectors. The second shared primer binding site is provided by a HRV 3C protease cleavage site located downstream of purification and solubility enhancing tags for tag removal. We have so far generated more than 30 different parallel expression vectors, and successfully cloned and expressed more than 250 genes with this vector series. There is no size restriction for gene insertion, clone efficiency is > 95% with clone numbers up to 200. The procedure is simple, fast, efficient and cost-effective. All expression vectors showed efficient expression of eGFP and different target proteins requested to be produced and purified at our Core Facility services. This new expression vector series allows efficient

  18. 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…

  19. An improved method for construction of directionally cloned cDNA libraries from microdissected cells.

    PubMed

    Peterson, L A; Brown, M R; Carlisle, A J; Kohn, E C; Liotta, L A; Emmert-Buck, M R; Krizman, D B

    1998-12-01

    Here, we developed an improved method for constructing microdissected cDNA libraries, based on strand-switching properties of reverse transcriptase, followed by PCR amplification with primers to mediate unidirectional insert cloning. Using RNA from microdissected ovarian carcinoma cells, we constructed a cDNA library consisting of 1.3 x 10(6) unidirectional recombinants with an average insert size of 500 bp. Single-pass sequencing of 100 clones with the T7 primer revealed 89 inserts derived from known genes, anonymous expressed sequence tags (ESTs), or novel sequences. Among these clones were known genes and ESTs previously found in cDNA libraries from bulk ovarian tissue RNA, sequences seen for the first time in an ovarian-derived library, and novel sequences not previously seen in any cDNA library. These results demonstrate a methodology for constructing quality cDNA libraries that are cloned in a unidirectional fashion, are complex and diverse, and reflect the tissue of origin.

  20. Successful completion of a semi-automated enzyme-free cloning method.

    PubMed

    Bonacci, Stefano; Buccato, Scilla; Maione, Domenico; Petracca, Roberto

    2016-09-01

    Nowadays, in scientific fields such as Structural Biology or Vaccinology, there is an increasing need of fast, effective and reproducible gene cloning and expression processes. Consequently, the implementation of robotic platforms enabling the automation of protocols is becoming a pressing demand. The main goal of our study was to set up a robotic platform devoted to the high-throughput automation of the polymerase incomplete primer extension cloning method, and to evaluate its efficiency compared to that achieved manually, by selecting a set of bacterial genes that were processed either in the automated platform (330) or manually (94). Here we show that we successfully set up a platform able to complete, with high efficiency, a wide range of molecular biology and biochemical steps. 329 gene targets (99 %) were effectively amplified using the automated procedure and 286 (87 %) of these PCR products were successfully cloned in expression vectors, with cloning success rates being higher for the automated protocols respect to the manual procedure (93.6 and 74.5 %, respectively).

  1. 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.

  2. 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.

  3. Innovative teaching methods for capacity building in knowledge translation

    PubMed Central

    2011-01-01

    Background In some current healthcare settings, there is a noticeable absence of national institutions committed to the synthesis and use of evidence in healthcare decision- and policy-making. This absence creates a need to broaden the responsibilities of healthcare providers to include knowledge brokering and advocacy in order to optimize knowledge translation to other stakeholders, especially policy-makers. However, this process requires practitioners and researchers to acquire certain types of knowledge and skills. This article introduces two innovative methods for capacity building in knowledge translation (KT). Methods During a workshop aimed at preparing 21 trainers in evidence-based medicine, two innovative methods were used: (1) debate and (2) a knowledge translation project (KTP). The main objective of the debates approach was to strengthen participants' critical thinking abilities by requiring them to search for and appraise evidence and defend their arguments. The KTP was used to introduce participants to the essential steps of knowledge translation and to suggest an extended role for healthcare practitioners, i.e., using evidence to manage not only individual patients but also to a community of patients. Participants' performances were assessed according to a pre-designed scheme. At the end of the workshop, participants' opinions and experiences with the innovative teaching methods were evaluated based on their answers to a questionnaire and the results of small-group discussions. Results The participants performed well in both the debate and KTP methods. During post-workshop group discussions, they indicated that the debate approach had added a new dimension to their evidence-based medicine skills by adding purpose and motivation. However, they felt that their performances would have been better if they had been offered practical demonstrations of how to conduct the debate. The participants indicated that the KTP enhanced their understanding of the

  4. QuickStep-Cloning: a sequence-independent, ligation-free method for rapid construction of recombinant plasmids.

    PubMed

    Jajesniak, Pawel; Wong, Tuck Seng

    2015-01-01

    Molecular cloning is an essential step in biological engineering. Methods involving megaprimer-based PCR of a whole plasmid are promising alternatives to the traditional restriction-ligation-based molecular cloning. Their widespread use, however, is hampered by some of their inherent characteristics, e.g., linear amplification, use of self-annealing megaprimers and difficulty with performing point insertion of DNA. These limitations result in low product yield and reduced flexibility in the design of a genetic construct. Here, we present a novel technique of directional cloning, which overcomes these problems yet retaining the simplicity of whole-plasmid amplification. QuickStep-Cloning utilizes asymmetric PCRs to create a megaprimer pair with 3'-overhangs, and hence, facilitates the subsequent exponential whole-plasmid amplification. QuickStep-Cloning generates nicked-circular plasmids, thereby permitting direct bacterial transformation without DNA ligation. It allows DNA fragment integration into any plasmid at any position, in an efficient, time- and cost-effective manner, without tedious intermediate DNA gel purification, modified oligonucleotides, specialty enzymes and ultra-competent cells. The method is compatible with competent E. coli cells prepared using the conventional calcium chloride method. QuickStep-Cloning expands the versatility of megaprimer-based cloning. It is an excellent addition to the cloning toolbox, for the benefit of protein engineers, metabolic engineers and synthetic biologists.

  5. An evaluation of the geographic method for recognizing innovations in nature, using zoo orangutans.

    PubMed

    Lehner, Stephan R; Burkart, Judith M; van Schaik, Carel P

    2010-04-01

    Innovation and social learning are the raw materials for traditions and culture. Of these two, innovation has received far less scrutiny, largely because of difficulties assessing the innovation status of behaviors. A recent attempt proposes recognition of innovations in natural populations based on assessment of the behavior's properties and its geographic and local prevalence. Here we examine the validity of this approach and the list of 43 potential innovations it generated for wild orangutans by extending the comparison to zoo orangutans. First, we created an inventory of the behavioral repertoire in the zoo population. Four of ten putative innovations recognized in the field and potentially present in captivity did not occur despite appropriate conditions, suggesting they are indeed innovations. Second, we experimentally produced relevant conditions to evaluate whether another five potential innovations could be elicited. Based on their continued absence or on their latencies relative to known behaviors, four of the potential innovations could be assessed as innovations and one as a modification. Because 53% of relevant innovations recognized in the field could be confirmed in this analysis, and another 27% assigned possible innovation status, we conclude that the geographic method for detecting innovation in the wild is valid. However, the experiments also yielded up to 13 additional innovations, suggesting that zoo orangutans are far more innovative than wild ones. We discuss the implications of this latter finding with regard to limiting factors for the expansion of cultural repertoires in wild orangutans.

  6. 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…

  7. 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…

  8. A universal cloning method based on yeast homologous recombination that is simple, efficient, and versatile

    PubMed Central

    Joska, Tammy M.; Mashruwala, Ameya; Boyd, Jeffrey M.; Belden, William J.

    2014-01-01

    Cloning by homologous recombination (HR) in Saccharomyces cerevisiae is an extremely efficient and cost-effective alternative to other methods of recombinant DNA technologies. Unfortunately, it is incompatible with all the various specialized plasmids currently used in microbiology and biomedical research laboratories, and is therefore, not widely adopted. In an effort to dramatically improve the versatility of yeast gap-repair cloning and make it compatible with any DNA plasmid, we demonstrate that by simply including a yeast-cloning cassette (YCC) that contains the 2-micron origin of replication (2 μm ori) and the ura3 gene for selection, multiple DNA fragments can be assembled into any DNA vector. We show this has almost unlimited potential by building a variety of plasmid for different uses including: recombinant protein production, epitope tagging, site-directed mutagenesis, and expression of fluorescent fusion proteins. We demonstrate the use in a variety of plasmids for use in microbial systems and even demonstrate it can be used in a vertebrate model. This method is remarkably simple and extremely efficient, plus it provides a significant cost saving over commercially available kits. PMID:24418681

  9. A universal cloning method based on yeast homologous recombination that is simple, efficient, and versatile.

    PubMed

    Joska, Tammy M; Mashruwala, Ameya; Boyd, Jeffrey M; Belden, William J

    2014-05-01

    Cloning by homologous recombination (HR) in Saccharomyces cerevisiae is an extremely efficient and cost-effective alternative to other methods of recombinant DNA technologies. Unfortunately, it is incompatible with all the various specialized plasmids currently used in microbiology and biomedical research laboratories, and is therefore, not widely adopted. In an effort to dramatically improve the versatility of yeast gap-repair cloning and make it compatible with any DNA plasmid, we demonstrate that by simply including a yeast-cloning cassette (YCC) that contains the 2-micron origin of replication (2μm ori) and the ura3 gene for selection, multiple DNA fragments can be assembled into any DNA vector. We show this has almost unlimited potential by building a variety of plasmid for different uses including: recombinant protein production, epitope tagging, site-directed mutagenesis, and expression of fluorescent fusion proteins. We demonstrate the use in a variety of plasmids for use in microbial systems and even demonstrate it can be used in a vertebrate model. This method is remarkably simple and extremely efficient, plus it provides a significant cost saving over commercially available kits.

  10. Influence of ovulation status, seasonality and embryo transfer method on development of cloned porcine embryos.

    PubMed

    Koo, O J; Kang, J T; Kwon, D K; Park, H J; Lee, B C

    2010-10-01

    To improve pig cloning efficiency, the present study evaluated the effect of ovulation status, seasonality and embryo transfer (ET) method on in vivo development of cloned porcine embryos. Cloned embryos were transferred to surrogate mothers on the same day of somatic cell nuclear transfer. In pre-ovulation stage (PO), pregnancy rate (PR) and delivery rate (DR) were 36.3% and 9.4%, respectively. In post-ovulation stage, 22.7% PR and 2.1% DR were recorded (both PR and DR are significantly higher in PO). When ET was performed during winter (December-February), spring (March-May), summer (June-August) and autumn (September-November), the PRs were 13.4%, 37.3%, 24.6% and 51.0%, while DRs were 0%, 12.7%, 4.3% and 7.8%, respectively. The highest PRs were recorded in autumn groups. However, DRs were significantly lower in autumn (7.8%) group compared with spring (12.7%) group. The PR was the lowest and no piglets were born in winter group, which might be because of the effect of low temperature during ET. To overcome the low PR in winter group, 0.25 ml straws were used for ET to minimize exposure time of embryos to ambient temperature. The straw ET group showed significantly higher PR in the winter group (23. 9%) compared with the conventional catheter-loading group (7.7%). We suggest that using PO recipient and ET in spring is the best condition for pig cloning. In addition, alternative method to reduce cold shock during ET in winter is necessary. © 2009 Blackwell Verlag GmbH.

  11. Sequence analysis of zein cDNAs obtained by an efficient mRNA cloning method.

    PubMed Central

    Heidecker, G; Messing, J

    1983-01-01

    A cDNA library was generated from mRNA isolated from the developing endosperm of W22 maize inbred. cDNA clones for zein, the maize storage protein family, were isolated and analyzed by DNA sequencing. The DNA sequences of four clones containing cDNA copies of mRNAs belonging to one zein subfamily were determined. The data support the following conclusions: a) genes encoding the larger of the two zein species contain eleven instead of nine repeat units within the coding sequence of the gene; b) transcription can be terminated at either of the two polyadenlation signals and c) transcription starts 31 basepairs downstream from the first T in the TATA box. To facilitate this analysis a new method for the construction of cDNA libraries was developed. The mRNA was annealed to linearized and oligo-dT tailed pUC9 plasmid DNA, which then primed synthesis of the first strand of the cDNA. Oligo-dG tails were added to the cDNA-plasmid molecules, which were then centrifuged through an alkaline sucrose gradient. The gradient step removed small molecules and separated the two cDNAs which were formerly attached to the same double stranded plasmid molecule. An excess of oligo-dC tailed denatured pUC9 DNA was added and the DNA was renatured under conditions that favor the circularization of monomers by the oligo-dC and oligo-dG tails. The oligo-dC tail served as primer for the synthesis of the second strand of the cDNA. The library was screened by colony hybridization using 32P-labelled cDNA and DNA from genomic zein clones as probes. We obtained 20,000 clones hybridizing total cDNA starting with 1 microgram of plasmid DNA and 1 microgram of mRNA. Images PMID:6688299

  12. Dog cloning with in vivo matured oocytes obtained using electric chemiluminescence immunoassay-predicted ovulation method.

    PubMed

    Lee, Seunghoon; Zhao, Minghui; No, Jingu; Nam, Yoonseok; Im, Gi-Sun; Hur, Tai-Young

    2017-01-01

    Radioactive immunoassay (RIA) is a traditional serum hormone assay method, but the application of the method in reproductive studies is limited by the associated radioactivity. The aim of present study was to evaluate the reliability of RIA and to compare its canine serum progesterone concentration determination accuracy to that of the electric chemiluminescence immunoassay (ECLI). In vivo matured oocytes were utilized for canine somatic cell nuclear transfer (SCNT), and serum progesterone levels were assessed to accurately determine ovulation and oocyte maturation. Canine serum progesterone concentrations during both proestrus and estrus were analyzed by RIA and ECLI to determine the ovulation day. Although both methods detected similar progesterone levels before ovulation, the mean progesterone concentration determined using ECLI was significantly higher than of RIA three days before ovulation. Following ovulation, oocytes were collected by surgery, and a lower percentage of mature oocytes were observed using ECLI (39%) as compared to RIA (67%) if 4-8ng/ml of progesterone were used for determination of ovulation. A high percentage of mature oocytes was observed using ECLI when 6-15 ng/mL of progesterone was used for ovulation determination. To determine whether ECLI could be used for canine cloning, six canines were selected as oocyte donors, and two puppies were obtained after SCNT and embryo transfer. In conclusion, compared to the traditional RIA method, the ECLI method is a safe and reliable method for canine cloning.

  13. Dog cloning with in vivo matured oocytes obtained using electric chemiluminescence immunoassay-predicted ovulation method

    PubMed Central

    No, Jingu; Nam, Yoonseok; Im, Gi-Sun; Hur, Tai-Young

    2017-01-01

    Radioactive immunoassay (RIA) is a traditional serum hormone assay method, but the application of the method in reproductive studies is limited by the associated radioactivity. The aim of present study was to evaluate the reliability of RIA and to compare its canine serum progesterone concentration determination accuracy to that of the electric chemiluminescence immunoassay (ECLI). In vivo matured oocytes were utilized for canine somatic cell nuclear transfer (SCNT), and serum progesterone levels were assessed to accurately determine ovulation and oocyte maturation. Canine serum progesterone concentrations during both proestrus and estrus were analyzed by RIA and ECLI to determine the ovulation day. Although both methods detected similar progesterone levels before ovulation, the mean progesterone concentration determined using ECLI was significantly higher than of RIA three days before ovulation. Following ovulation, oocytes were collected by surgery, and a lower percentage of mature oocytes were observed using ECLI (39%) as compared to RIA (67%) if 4-8ng/ml of progesterone were used for determination of ovulation. A high percentage of mature oocytes was observed using ECLI when 6–15 ng/mL of progesterone was used for ovulation determination. To determine whether ECLI could be used for canine cloning, six canines were selected as oocyte donors, and two puppies were obtained after SCNT and embryo transfer. In conclusion, compared to the traditional RIA method, the ECLI method is a safe and reliable method for canine cloning. PMID:28288197

  14. 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…

  15. 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…

  16. Innovative methods to promote optics at school level

    NASA Astrophysics Data System (ADS)

    Garg, Amit; Bharadwaj, Sadashiv Raj; Kachru, Priyanka; Dutta, Subhajit Kumar; Singh, Shatakshi

    2016-09-01

    The conventional pedagogical techniques in Indian schools are predominantly limited to coursework and involve just necessary laboratory work. It lacks in inculcating the aptitude for practical applications relevant to the coursework amongst the students. This results in a major setback for the students as their mental approach remains constrained to books, where on the contrary, the future depends only on technologies and applications. Considering this, under Optics Outreach Education Program 2014, University of Delhi at Acharya Narendra Dev College SPIE Student Chapter conducted workshops in 22 schools of Delhi including various Government schools as well. The workshop zeroed into the concepts of optics and various innovative techniques developed by our Student chapter members to embark a new method of teaching.

  17. An innovative method of providing total breaker failure protection

    SciTech Connect

    Stringer, N.T.; Waser, D.

    1995-12-31

    Breaker failure relaying has been generally achieved through the use of a current monitoring relay to determine whether current continues to flow into a fault after a breaker has been instructed to interrupt the circuit. If current continues to flow after a predefined period of time, the circuit breaker is considered to have failed. Steps must then be taken to trip the next set of upstream breakers in the power system to remove the faulted circuit and prevent system damage. However, with industrial power systems, this may be the utility`s breakers on the feeding transmission line. Regardless, breaker failure schemes must be designed to isolate both the faulted circuit and the failed circuit breaker. This paper discusses a new and innovative method of protecting a circuit breaker from the failure described above, plus other failures that go unprotected with conventional schemes, thus providing ``total`` breaker failure protection.

  18. An innovative method of providing total breaker failure protection

    SciTech Connect

    Stringer, N.T.; Waser, D.

    1996-09-01

    Breaker failure relaying has been generally achieved through the use of a current monitoring relay to determine whether current continues to flow into a fault after a breaker has been instructed to interrupt the circuit. If current continues to flow after a predefined period of time, the circuit breaker is considered to have failed. Steps must then be taken to trip the next set of upstream breakers in the power system to remove the faulted circuit and prevent system damage. However, with industrial power systems, this may be the utility`s breakers on the feeding transmission line. Regardless, breaker failure schemes must be designed to isolate both the faulted circuit and the failed circuit breaker. This paper discusses a new and innovative method of protecting a circuit breaker from the failure described above, plus other failures that go unprotected with conventional schemes, this providing total breaker failure protection.

  19. Efficiency of two enucleation methods connected to handmade cloning to produce transgenic porcine embryos.

    PubMed

    Li, J; Villemoes, K; Zhang, Y; Du, Y; Kragh, P M; Purup, S; Xue, Q; Pedersen, A M; Jørgensen, A L; Jakobsen, J E; Bolund, L; Yang, H; Vajta, G

    2009-02-01

    The purpose of our work was to establish an efficient-oriented enucleation method to produce transgenic embryos with handmade cloning (HMC). After 41-42 h oocytes maturation, the oocytes were further cultured with or without 0.4 microg/ml demecolcine for 45 min [chemically assisted handmade enucleation (CAHE) group vs polar body (PB) oriented handmade enucleation (OHE) group respectively]. After removal of the cumulus cells and partial digestion of the zona pellucida, oocytes with visible extrusion cones and/or polar bodies attached to the surface were subjected to oriented bisection. Putative cytoplasts without extrusion cones or PB were selected as recipients. Two cytoplasts were electrofused with one transgenic fibroblasts expressing green fluorescent protein (GFP), while non-transgenic fibroblasts were used as controls. Reconstructed embryos were cultured in Well of Wells (WOWs) with porcine zygote medium 3 (PZM-3) after activation. Cleavage and blastocyst rates were registered on day 2 and day 7 of in vitro culture respectively. Meanwhile, the total blastocyst cell number was counted on day 7. We found that the difference was only observed between blastocyst rates (38.6 +/- 2% vs 48.1 +/- 3%) of cloned embryos with GFP transgenic fibroblast cells after CAHE vs OHE. With adjusted time-lapse for zonae-free cloned embryos cultured in WOWs with PZM-3, it was obvious that in vitro developmental competence after CAHE was compromised when compared with the OHE method. OHE enucleation method seems to be a potential superior alternative method used for somatic cell nuclear transfer (SCNT) with transgenic fibroblast cells.

  20. An innovative lossless compression method for discrete-color images.

    PubMed

    Alzahir, Saif; Borici, Arber

    2015-01-01

    In this paper, we present an innovative method for lossless compression of discrete-color images, such as map images, graphics, GIS, as well as binary images. This method comprises two main components. The first is a fixed-size codebook encompassing 8×8 bit blocks of two-tone data along with their corresponding Huffman codes and their relative probabilities of occurrence. The probabilities were obtained from a very large set of discrete color images which are also used for arithmetic coding. The second component is the row-column reduction coding, which will encode those blocks that are not in the codebook. The proposed method has been successfully applied on two major image categories: 1) images with a predetermined number of discrete colors, such as digital maps, graphs, and GIS images and 2) binary images. The results show that our method compresses images from both categories (discrete color and binary images) with 90% in most case and higher than the JBIG-2 by 5%-20% for binary images, and by 2%-6.3% for discrete color images on average.

  1. A genetic method for generating Drosophila eyes composed exclusively of mitotic clones of a single genotype.

    PubMed Central

    Stowers, R S; Schwarz, T L

    1999-01-01

    The genetic analysis of a gene at a late developmental stage can be impeded if the gene is required at an earlier developmental stage. The construction of mosaic animals, particularly in Drosophila, has been a means to overcome this obstacle. However, the phenotypic analysis of mitotic clones is often complicated because standard methods for generating mitotic clones render mosaic tissues that are a composite of both mutant and phenotypically normal cells. We describe here a genetic method (called EGUF/hid) that uses both the GAL4/UAS and FLP/FRT systems to overcome this limitation for the Drosophila eye by producing genetically mosaic flies that are otherwise heterozygous but in which the eye is composed exclusively of cells homozygous for one of the five major chromosome arms. These eyes are nearly wild type in size, morphology, and physiology. Applications of this genetic method include phenotypic analysis of existing mutations and F(1) genetic screens to identify as yet unknown genes involved in the biology of the fly eye. We illustrate the utility of the method by applying it to lethal mutations in the synaptic transmission genes synaptotagmin and syntaxin. PMID:10430588

  2. A method for construction, cloning and expression of intron-less gene from unannotated genomic DNA.

    PubMed

    Agrawal, Vineet; Gupta, Bharti; Banerjee, Uttam Chand; Roy, Nilanjan

    2008-11-01

    The present century has witnessed an unprecedented rise in genome sequences owing to various genome-sequencing programs. However, the same has not been replicated with cDNA or expressed sequence tags (ESTs). Hence, prediction of protein coding sequence of genes from this enormous collection of genomic sequences presents a significant challenge. While robust high throughput methods of cloning and expression could be used to meet protein requirements, lack of intron information creates a bottleneck. Computational programs designed for recognizing intron-exon boundaries for a particular organism or group of organisms have their own limitations. Keeping this in view, we describe here a method for construction of intron-less gene from genomic DNA in the absence of cDNA/EST information and organism-specific gene prediction program. The method outlined is a sequential application of bioinformatics to predict correct intron-exon boundaries and splicing by overlap extension PCR for spliced gene synthesis. The gene construct so obtained can then be cloned for protein expression. The method is simple and can be used for any eukaryotic gene expression.

  3. Hot Fusion: An Efficient Method to Clone Multiple DNA Fragments as Well as Inverted Repeats without Ligase

    PubMed Central

    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%. PMID:25551825

  4. 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.

  5. Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos.

    PubMed

    Ma, LiBing; Liu, XiYu; Wang, FengMei; He, XiaoYing; Chen, Shan; Li, WenDa

    2015-01-01

    It was proposed that arresting nuclear donor cells in G0/G1 phase facilitates the development of embryos that are derived from somatic cell nuclear transfer (SCNT). Full confluency or serum starvation is commonly used to arrest in vitro cultured somatic cells in G0/G1 phase. However, it is controversial as to whether these two methods have the same efficiency in arresting somatic cells in G0/G1 phase. Moreover, it is unclear whether the cloned embryos have comparable developmental ability after somatic cells are subjected to one of these methods and then used as nuclear donors in SCNT. In the present study, in vitro cultured sheep skin fibroblasts were divided into four groups: (1) cultured to 70-80% confluency (control group), (2) cultured to full confluency, (3) starved in low serum medium for 4 d, or (4) cultured to full confluency and then further starved for 4 d. Flow cytometry was used to assay the percentage of fibroblasts in G0/G1 phase, and cell counting was used to assay the viability of the fibroblasts. Then, real-time reverse transcription PCR was used to determine the levels of expression of several cell cycle-related genes. Subsequently, the four groups of fibroblasts were separately used as nuclear donors in SCNT, and the developmental ability and the quality of the cloned embryos were compared. The results showed that the percentage of fibroblasts in G0/G1 phase, the viability of fibroblasts, and the expression levels of cell cycle-related genes was different among the four groups of fibroblasts. Moreover, the quality of the cloned embryos was comparable after these four groups of fibroblasts were separately used as nuclear donors in SCNT. However, cloned embryos derived from fibroblasts that were cultured to full confluency combined with serum starvation had the highest developmental ability. The results of the present study indicate that there are synergistic effects of full confluency and serum starvation on arresting fibroblasts in G0/G1 phase

  6. 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

  7. 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.

  8. WasteWise Resource Management: Innovative Solid Waste Contracting Methods

    EPA Pesticide Factsheets

    Resource management is an innovative contractual partnership between a waste-generating organization and a qualified contractor that changes the nature of current disposal services to support waste minimization and recycling.

  9. 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 ...

  10. 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.

  11. 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

  12. Development of a Multilocus Sequence Typing Method for Analysis of Listeria monocytogenes Clones

    PubMed Central

    Salcedo, C.; Arreaza, L.; Alcalá, B.; de la Fuente, L.; Vázquez, J. A.

    2003-01-01

    This study is a first step in the development of multilocus sequence typing (MLST) method for Listeria monocytogenes. Nine housekeeping genes were analyzed in a set of 62 strains isolated from different sources and geographic locations in Spain. These strains were previously characterized by pulsed-field gel electrophoresis (PFGE). Because of low diversity, two loci were discarded from the study. The sequence analysis of the seven remaining genes showed 29 different allelic combinations, with 22 of them represented by only one strain. The results of this sequence analysis were generally consistent with those of PFGE. Because MLST allows the easy comparison and exchange of results obtained in different laboratories, the future application of this new molecular method could be a useful tool for the listeriosis surveillance systems that will allow the identification and distribution of analysis of L. monocytogenes clones in the environment. PMID:12574278

  13. Report of Symposium: Applications of Advanced and Innovative Computational Methods to Defense Science and Engineering.

    DTIC Science & Technology

    1994-11-01

    Problem Comments Machine and Software 1 Computational Fluid Dynamics PDE, FEM Turbulence Mesh Generation SIMD MIMD for irregular, adaptive...COVERED Final Report of Symposium-Applications of Advanced and Innovative Computational Methods to Defense Science and Engineering 6. AUTHOR(S...UNCLASSIFIED Copy 15 of 122 copies IDA DOCUMENT D-1657 REPORT OF SYMPOSIUM APPLICATIONS OF ADVANCED AND INNOVATIVE COMPUTATIONAL METHODS TO

  14. Data cloning: easy maximum likelihood estimation for complex ecological models using Bayesian Markov chain Monte Carlo methods.

    PubMed

    Lele, Subhash R; Dennis, Brian; Lutscher, Frithjof

    2007-07-01

    We introduce a new statistical computing method, called data cloning, to calculate maximum likelihood estimates and their standard errors for complex ecological models. Although the method uses the Bayesian framework and exploits the computational simplicity of the Markov chain Monte Carlo (MCMC) algorithms, it provides valid frequentist inferences such as the maximum likelihood estimates and their standard errors. The inferences are completely invariant to the choice of the prior distributions and therefore avoid the inherent subjectivity of the Bayesian approach. The data cloning method is easily implemented using standard MCMC software. Data cloning is particularly useful for analysing ecological situations in which hierarchical statistical models, such as state-space models and mixed effects models, are appropriate. We illustrate the method by fitting two nonlinear population dynamics models to data in the presence of process and observation noise.

  15. 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.

  16. Rapid Restriction Enzyme-Free Cloning of PCR Products: A High-Throughput Method Applicable for Library Construction

    PubMed Central

    Chaudhary, Vijay K.; Das, Shilpi; Kaur, Charanpreet; Grover, Payal; Gupta, Amita

    2014-01-01

    Herein, we describe a novel cloning strategy for PCR-amplified DNA which employs the type IIs restriction endonuclease BsaI to create a linearized vector with four base-long 5′-overhangs, and T4 DNA polymerase treatment of the insert in presence of a single dNTP to create vector-compatible four base-long overhangs. Notably, the insert preparation does not require any restriction enzyme treatment. The BsaI sites in the vector are oriented in such a manner that upon digestion with BsaI, a stuffer sequence along with both BsaI recognition sequences is removed. The sequence of the four base-long overhangs produced by BsaI cleavage were designed to be non-palindromic, non-compatible to each other. Therefore, only ligation of an insert carrying compatible ends allows directional cloning of the insert to the vector to generate a recombinant without recreating the BsaI sites. We also developed rapid protocols for insert preparation and cloning, by which the entire process from PCR to transformation can be completed in 6–8 h and DNA fragments ranging in size from 200 to 2200 bp can be cloned with equal efficiencies. One protocol uses a single tube for insert preparation if amplification is performed using polymerases with low 3′-exonuclease activity. The other protocol is compatible with any thermostable polymerase, including those with high 3′-exonuclease activity, and does not significantly increase the time required for cloning. The suitability of this method for high-throughput cloning was demonstrated by cloning batches of 24 PCR products with nearly 100% efficiency. The cloning strategy is also suitable for high efficiency cloning and was used to construct large libraries comprising more than 108 clones/µg vector. Additionally, based on this strategy, a variety of vectors were constructed for the expression of proteins in E. coli, enabling large number of different clones to be rapidly generated. PMID:25360695

  17. Method for cloning in vivo targets of the Egr-1 transcription factor.

    PubMed

    de Belle, I; Mercola, D; Adamson, E D

    2000-07-01

    A methodology is described that allows the in vivo trapping of transcription factors to their target regulatory elements in multiple genes simultaneously. Cross-linking using formaldehyde is the first of several steps to isolate, purify, clone and characterize multiple gene promoter DNA fragments. The example that we use indicates that the TGF beta 1 gene is a direct target induced by Egr-1 in HT1080 cells that express constitutive Egr-1, thus explaining the growth retardation that follows Egr-1 expression. The genes identified using this procedure reflect the specific activities of Egr-1 at that moment in the cell and provide a method for confirmation of genes that are the direct targets of Egr-1 action.

  18. New Research: Changing Schools, Innovation Up Close, Improving Research Methods.

    ERIC Educational Resources Information Center

    Hood, Paul, Ed.

    1983-01-01

    This issue of Research and Educational Practice Improvement Notes discusses three recent studies on schooling: (1) "Changing Our Schools; The Realities," by Kenneth Tye; (2) "Innovation Up Close: A Field Study in Twelve School Settings," Volume Four of a study conducted by David P. Crandall and associates; and (3) "Methodological Research on…

  19. 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.…

  20. New Research: Changing Schools, Innovation Up Close, Improving Research Methods.

    ERIC Educational Resources Information Center

    Hood, Paul, Ed.

    1983-01-01

    This issue of Research and Educational Practice Improvement Notes discusses three recent studies on schooling: (1) "Changing Our Schools; The Realities," by Kenneth Tye; (2) "Innovation Up Close: A Field Study in Twelve School Settings," Volume Four of a study conducted by David P. Crandall and associates; and (3) "Methodological Research on…

  1. 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.

  2. Methods of rapid, early selection of poplar clones for maximum yield potential: a manual of procedures.

    Treesearch

    USDA FS

    1982-01-01

    Instructions, illustrated with examples and experimental results, are given for the controlled-environment propagation and selection of poplar clones. Greenhouse and growth-room culture of poplar stock plants and scions are described, and statistical techniques for discriminating among clones on the basis of growth variables are emphasized.

  3. 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

  4. Innovative Methods for Corn Stover Collecting, Handling, Storing and Transporting

    DTIC Science & Technology

    2004-04-01

    stover feedstock to become a reality for large biorefineries , innovations are needed between the field and delivery to the processor in three areas: o...rail transport helps close the gap between petroleum refineries averaging over 100,000 barrels per day, Table D. Table D Biorefinery Feedstock...Circles with 50 mile Collection Radius Conclusions • Biorefineries with wet processes are most likely to use feedstock supplied by unit trains

  5. A clone screening method using mRNA levels to determine specific productivity and product quality for monoclonal antibodies.

    PubMed

    Lee, Christina J; Seth, Gargi; Tsukuda, Joni; Hamilton, Robert W

    2009-03-01

    To meet increasing demands for efficient and streamlined production processes of therapeutic antibodies, improved methods of screening clones are required. In this article, we examined the potential of using antibody transcript levels as criteria for clone screening. We evaluated the QuantiGene Plex, a commercially available, high-throughput assay for simultaneously measuring multiple transcripts from cell lysate. Using the development of stable Chinese hamster ovary cell lines as examples, we investigated the relationship between transcript and antibody levels through several rounds of screening. First, we observed that measured heavy chain transcript levels are generally correlated with specific productivity, enabling the identification of high-producing clones from mRNA. Second, we observed that low ratios (< 1.5) of light to heavy chain transcript levels may be indicative of high antibody aggregation levels, allowing for the rapid identification and elimination of clones of questionable product quality. Therefore, an efficient process of identifying high-producing clones of desirable product quality is possible by using QuantiGene Plex assay to measure antibody transcript levels.

  6. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Creation of DNA overhangs by using modified DNA overhang cloning method.

    PubMed

    Li, Wei; Gao, Feng

    2007-06-01

    DNA overhang cloning (DOC) method was invented by Kevin A. Jarrell and colleagues to create DNA overhangs at the ends of polymerase chain reaction (PCR) fragments. To increase the ligation products of DOC and make it possible to join the ligation products together to yield a larger DNA fragment without amplification by using the secondary PCR, we modified the original method by lengthening RNA-DNA hybrids to 8 bp, using RNase H to digest RNA chains, and so on. We found that the ligation of PCR fragments with DNA overhangs could be improved significantly by using the modified DOC. The ligation products of the modified DOC were rich enough to be separated by using agarose gel and joined together to yield a larger DNA fragment without amplification by using the secondary PCR. The modified DOC is simpler and much more effective than the original one. The creation of DNA overhangs by using complementary DNA oligonucleotides, T4 ligase, and RNase H is simpler and cheaper than that by using reverse transcriptase and RNase H.

  8. Comparison of molecular typing methods for characterization of Staphylococcus epidermidis: proposal for clone definition.

    PubMed

    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.

  9. An alternative method for cDNA cloning from surrogate eukaryotic cells transfected with the corresponding genomic DNA.

    PubMed

    Hu, Lin-Yong; Cui, Chen-Chen; Song, Yu-Jie; Wang, Xiang-Guo; Jin, Ya-Ping; Wang, Ai-Hua; Zhang, Yong

    2012-07-01

    cDNA is widely used in gene function elucidation and/or transgenics research but often suitable tissues or cells from which to isolate mRNA for reverse transcription are unavailable. Here, an alternative method for cDNA cloning is described and tested by cloning the cDNA of human LALBA (human alpha-lactalbumin) from genomic DNA. First, genomic DNA containing all of the coding exons was cloned from human peripheral blood and inserted into a eukaryotic expression vector. Next, by delivering the plasmids into either 293T or fibroblast cells, surrogate cells were constructed. Finally, the total RNA was extracted from the surrogate cells and cDNA was obtained by RT-PCR. The human LALBA cDNA that was obtained was compared with the corresponding mRNA published in GenBank. The comparison showed that the two sequences were identical. The novel method for cDNA cloning from surrogate eukaryotic cells described here uses well-established techniques that are feasible and simple to use. We anticipate that this alternative method will have widespread applications.

  10. 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.

  11. 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

  12. An innovative cloning platform enables large-scale production and maturation of an oxygen-tolerant [NiFe]-hydrogenase from Cupriavidus necator in Escherichia coli.

    PubMed

    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

  13. 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…

  14. 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…

  15. 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…

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

    PubMed

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

    2014-02-01

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

  17. Combining the polymerase incomplete primer extension method for cloning and mutagenesis with microscreening to accelerate structural genomics efforts.

    PubMed

    Klock, Heath E; Koesema, Eric J; Knuth, Mark W; Lesley, Scott A

    2008-05-01

    Successful protein expression, purification, and crystallization for challenging targets typically requires evaluation of a multitude of expression constructs. Often many iterations of truncations and point mutations are required to identify a suitable derivative for recombinant expression. Making and characterizing these variants is a significant barrier to success. We have developed a rapid and efficient cloning process and combined it with a protein microscreening approach to characterize protein suitability for structural studies. The Polymerase Incomplete Primer Extension (PIPE) cloning method was used to rapidly clone 448 protein targets and then to generate 2143 truncations from 96 targets with minimal effort. Proteins were expressed, purified, and characterized via a microscreening protocol, which incorporates protein quantification, liquid chromatography mass spectrometry and analytical size exclusion chromatography (AnSEC) to evaluate suitability of the protein products for X-ray crystallography. The results suggest that selecting expression constructs for crystal trials based primarily on expression solubility is insufficient. Instead, AnSEC scoring as a measure of protein polydispersity was found to be predictive of ultimate structure determination success and essential for identifying appropriate boundaries for truncation series. Overall structure determination success was increased by at least 38% by applying this combined PIPE cloning and microscreening approach to recalcitrant targets.

  18. Easy assessment of ES cell clone potency for chimeric development and germ-line competency by an optimized aggregation method.

    PubMed

    Kondoh, G; Yamamoto, Y; Yoshida, K; Suzuki, Y; Osuka, S; Nakano, Y; Morita, T; Takeda, J

    1999-05-13

    Production of germ-line competent chimeric mice from embryonic stem (ES) cells is an inevitable step in establishing gene-manipulated mouse lineages. A common method used for creating chimeric mice is the injection of ES cells into the blastocoelic cavity (blastocyst injection). The aggregation method is an alternative way to introduce ES cells to the host embryo which is less difficult than blastocyst injection. Here we re-examined the condition of embryo-ES cell coculture on the aggregation method and found improvement of germ-line competent chimeric production by a simple modification of the coculture medium. Moreover, R1 ES cell and its 10 gene-manipulated subclones were tested by this method. Although all ES cell clones showed good morphology and a normal karyotype, the efficiency of chimeric development and germ-line transmission varied among clones and were classified into three grades according to germ-line competency. In the first group (class A), both the incidence of chimera with high ES cell contribution and the rate of germ-line transmission were fairly high. Germ-line competent chimeras were obtained but with rather low efficiency in the second group (class B), while another group (class C) showed an absence of high ES cell-contributed chimeras and no germ-line transmission. These results suggest the usefulness of this modified aggregation method to predict the potency of ES cell clones for germ-line competency.

  19. Room for improvement? Leadership, innovation culture and uptake of quality improvement methods in general practice.

    PubMed

    Apekey, Tanefa A; McSorley, Gerry; Tilling, Michelle; Siriwardena, A Niroshan

    2011-04-01

    Leadership and innovation are currently seen as essential elements for the development and maintenance of high-quality care. Little is known about the relationship between leadership and culture of innovation and the extent to which quality improvement methods are used in general practice. This study aimed to assess the relationship between leadership behaviour, culture of innovation and adoption of quality improvement methods in general practice. Self-administered postal questionnaires were sent to general practitioner quality improvement leads in one county in the UK between June and December 2007. The questionnaire consisted of background information, a 12-item scale to assess leadership behaviour, a seven-dimension self-rating scale for culture of innovation and questions on current use of quality improvement tools and techniques. Sixty-three completed questionnaires (62%) were returned. Leadership behaviours were not commonly reported. Most practices reported a positive culture of innovation, featuring relationship most strongly, followed by targets and information but rated lower on other dimensions of rewards, risk and resources. There was a significant positive correlation between leadership behaviour and the culture of innovation (r = 0.57; P < 0.001). Apart from clinical audit and significant event analysis, quality improvement methods were not adopted by most participating practices. Leadership behaviours were infrequently reported and this was associated with a limited culture of innovation in participating general practices. There was little use of quality improvement methods beyond clinical and significant event audit. Practices need support to enhance leadership skills, encourage innovation and develop quality improvement skills if improvements in health care are to accelerate. © 2010 Blackwell Publishing Ltd.

  20. Student Characteristics, Prior Experiences, and the Perception of Mixed Methods as an Innovation

    ERIC Educational Resources Information Center

    Brown, Sydney E.

    2014-01-01

    There are persistent challenges to teaching mixed methods and innovative solutions are sought in order to address the needs of an increasingly diverse global audience seeking mixed methods instruction. This mixed methods study was conducted to gain insights to course design by more fully understanding the relationships among graduate student…

  1. Application of Innovative P&E Method at Technical Universities in Slovakia

    ERIC Educational Resources Information Center

    Nemec, Miroslav; Krišták, Luboš; Hockicko, Peter; Danihelová, Zuzana; Velmovská, Klára

    2017-01-01

    The paper deals with innovative teaching methods at universities. The result of this effort is the interactive P&E method, whose main idea is the interactive work with students while solving problem tasks. The main aim of the given method is to change the students' position, by means of experiment analyses and qualitative tasks, from a passive…

  2. Student Characteristics, Prior Experiences, and the Perception of Mixed Methods as an Innovation

    ERIC Educational Resources Information Center

    Brown, Sydney E.

    2014-01-01

    There are persistent challenges to teaching mixed methods and innovative solutions are sought in order to address the needs of an increasingly diverse global audience seeking mixed methods instruction. This mixed methods study was conducted to gain insights to course design by more fully understanding the relationships among graduate student…

  3. 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.

  4. 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

  5. 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

  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.

  7. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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,…

  12. 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,…

  13. 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.…

  14. 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.

  15. Feasibility study for an innovative industrial red mud utilisation method.

    PubMed

    Kounalakis, Petros; Aravossis, Konstantinos; Karayianni, ChS

    2016-02-01

    Red mud is a high volume industrial waste, and its management poses a unique challenge. For the utilisation of red mud, an economical, energy saving, environmental friendly and widely applicable method has been found. The proposed novel method is purely chemical, and achieves the recovery of all the oxides contained in red mud totally and transforms them into high value added products. The present work shows that an investment in an industrial plant, treating red mud and turning a toxic industrial waste in commercial products, is safe and viable. © The Author(s) 2015.

  16. Innovative Teaching Practice: Traditional and Alternative Methods (Challenges and Implications)

    ERIC Educational Resources Information Center

    Nurutdinova, Aida R.; Perchatkina, Veronika G.; Zinatullina, Liliya M.; Zubkova, Guzel I.; Galeeva, Farida T.

    2016-01-01

    The relevance of the present issue is caused be the strong need in alternative methods of learning foreign language and the need in language training and retraining for the modern professionals. The aim of the article is to identify the basic techniques and skills in using various modern techniques in the context of modern educational tasks. The…

  17. Innovative Teaching Methods for the 21st Century Students

    ERIC Educational Resources Information Center

    Deepak

    2005-01-01

    Futurum in Sweden is described here as a school that does not have any classrooms, there is nothing like a timetable, no school bells can be heard, the students do not have any school bags, and teachers are without desks. Futurum is a radically new school that merges creative architecture with modern learning and teaching methods. This article…

  18. Chemical Poker: An Innovative Testing Method Using Elements of Randomness.

    ERIC Educational Resources Information Center

    Benvenuto, Mark A.; Ferruzzi, Arthur

    2002-01-01

    Introduces a testing method for general and organic chemistry in which students receive prepared randomized cards with a portion of a question on them to complete and then use them to answer specific test questions. Discusses the effects of this technique on each individual student's learning process. (KHR)

  19. Training in Reading Skills: An Innovative Method from College Instruction.

    ERIC Educational Resources Information Center

    Shaw, Victor N.

    2001-01-01

    Presents a method that involves the execution of a make-the-quiz exercise to inspire students to read for major facts and ideas in academic publications. Concludes that students take the instructor's perspective, internalize class performance standards, and hence voluntarily follow those standards in their attitudes and behavior toward learning…

  20. Innovative Teaching Methods for the 21st Century Students

    ERIC Educational Resources Information Center

    Deepak

    2005-01-01

    Futurum in Sweden is described here as a school that does not have any classrooms, there is nothing like a timetable, no school bells can be heard, the students do not have any school bags, and teachers are without desks. Futurum is a radically new school that merges creative architecture with modern learning and teaching methods. This article…

  1. 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.

  2. [Innovations in cardiology. We are too fast with new methods].

    PubMed

    Diegeler, A

    2016-03-01

    Cardiology is rapidly developing on many levels. New treatment methods are introduced at ever decreasing intervals. Against the background of economization of other areas in medicine, dangers are lurking here for patients if safety, usefulness and sustainability of the treatment methods cannot be sufficiently proven. The German Federal Ministry of Health (Bundesministerium für Gesundheit, BMG) aims to adjust the regulatory framework for the approval of new medical products to the legal requirements of the European Union. With the establishment of the Institute for Quality Assessment and Transparency in Health Care (Institut für Qualitätssicherung und Transparenz im Gesundheitswesen, IQTIG) more precise quality controls should be carried out. Implantation registers will be soon implemented and the routinely performed quality control of different interventions will be coordinated across different healthcare sectors in order to achieve a better understanding of long-term results. Medicine in general and the safety of patients in particular, ultimately benefit from more stringent controls, neutrality and transparency in the assessment of new methods.

  3. Innovative machining method on CVD diamond thin film

    NASA Astrophysics Data System (ADS)

    Tsai, Hung-Yin; Cheng, Chih-Yung; Liu, Pin-Yin; Wu, Tung-Chuan

    2002-11-01

    Chemical vapor deposition (CVD) diamond has outstanding properties, including low thermal expansion, high chemical resistance and high acoustic propagation and has been widely used in optical, electrical, mechanical, chemical and thermal applications. Since synthesized diamond film results in large surface roughness, the surface treatment or polishing should be applied to expand the applications. Although reducing surface roughness by mechanical and etching methods have been investigated, the cost on the complicated equipment and on the long processing time is the most particular important issue. A new method is developed in the present study to approach the smooth surface by a catalytic grinding wheel. As grinding, the catalytic reaction occurs at the contact area between the grinding wheel and the diamond surface, and sp3 structure of diamond can be converted to sp2 structure with lower bonding energy; therefore, the lower surface roughness. Consequently, the average surface roughness is extremely improved from 230 nm to 20 nm, and the processing time can be shortened 10 times more than conventional methods, either lapping, or chemical assisted lapping.

  4. 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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Innovative antimicrobial susceptibility testing method using surface plasmon resonance.

    PubMed

    Chiang, Ya-Ling; Lin, Chi-Hung; Yen, Muh-Yong; Su, Yuan-Deng; Chen, Shean-Jen; Chen, How-Foo

    2009-03-15

    Utilizing the ultra sensitivity of surface plasmon resonance (SPR) biosensor to examine drug resistance of bacteria was studied in this research. Susceptible and resistant strains of Escherichia coli JM109 to ampicillin and those of Staphylococcus epidermidis to tetracycline, served as a blind test, were examined. The bacteria adhered on the Au thin film was treated by the injection of antibiotic flow. The optical property change of the bacteria responded to antibiotics were recorded through SPR mechanism. As a result, the susceptible strain of E. coli generally revealed more than three times of SPR angle shift when compared to the resistant one; the susceptible strain of S. epidermidis revealed irregular SPR angle shift while the resistant strain kept the SPR angle almost unchanged. The new SPR method took less than 2h of antibiotic treatment time to complete the antimicrobial susceptibility test. Different from conventional applications of SPR, specific antibodies is not required in this method. As compared to the conventional assays, Kirby-Bauer disk diffusion and variations of broth microdilution usually take 1 day to weeks to issue the report. Using this SPR assay can greatly reduce the waiting period for laboratory tests, and can therefore benefit patients who need proper antibiotic treatments to control bacterial infections. The sensitivity of the SPR biosensor built for the application is around 1.4 x 10(-4) on the refractive index.

  6. Slurm: An innovative Particle-in-Cell Method for Magnetohydrodynamics

    NASA Astrophysics Data System (ADS)

    Bacchini, Fabio; Olshevsky, Vyacheslav; Lapenta, Giovanni

    2016-10-01

    We present a new Particle-in-Cell method for plasma simulations. This is based on the original algorithm of FLIP-MHD, which uses a Lagrangian formulation of the macroscopic equations. A finite-difference approximation of the equations of motion is solved on a fixed (non-moving) grid, while convection of the quantities is modelled with the support of Lagrangian particles. Interpolation with first-order b-splines is used to project the conserved quantities from particles to the grid and back. In this work, we introduce two modifications of the original scheme. A particle volume evolution procedure is adopted to reduce the computational error, based on the Material Point Method for solid mechanics. The additional step introduces little to none computational diffusion and efficiently suppresses the so-called ringing instability, allowing the use of explicit time differencing. Furthermore, we eliminate the need for a Poisson solver in the magnetic field computation with the use of a vector potential. The vector potential evolution is modelled with a moving grid and interpolated to the fixed grid points to obtain a solenoidal magnetic field. The results of a number of HD and MHD tests show good agreement with the reference solutions and rather fast time and space convergence. Air Force Office of Scientific Research, Air Force Materiel Command, USAF under Award No. FA9550-14-1-0375. European Community's Seventh Framework Programme (FP7/2007-2013) via the DEEP-ER project under Grant Agreement No. 610476.

  7. TQWT and WDGA: innovative methods for ground roll attenuation

    NASA Astrophysics Data System (ADS)

    Goudarzi, Alireza; Riahi, Mohammad Ali

    2013-12-01

    The existence of noise in seismic data has an undesirable effect on seismic interpretations. An important type of coherent noise is ground roll, in which time and frequency overlap with useful seismic signals. The tunable Q-factor wavelet transform is a new discrete wavelet transform method and is considered as a new tool for ground roll attenuation. The simple tunable Q-factor wavelet transform by using a single wavelet with explicit oscillatory behavior is utilized for each trace. The enhanced tunable Q-factor wavelet transform (for seismic signals), which benefits from several mother wavelets with distinct Q-factors (for each part of a signal), is presented here and it is shown that it can adjust reliably to the natural non-stationary behavior of a seismic signal in time and space. Random noise has an undesirable effect on Q-factor ranges, with extreme consequences. Wavelet domain ground roll analysis is used as a substitute approach to thresholding for ground roll attenuation. The simple tunable Q-factor wavelet transform and the enhanced tunable Q-factor wavelet transform are compared with each other and with the f-k method by using synthetic and real data samples.

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

    PubMed

    Gagne, Joshua J; Thompson, Lauren; O'Keefe, Kelly; Kesselheim, Aaron S

    2014-11-24

    To examine methods for generating evidence on health outcomes in patients with rare diseases. Methodological review of existing literature. 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. 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. 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. 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. © Gagne et al 2014.

  9. Facile and innovative method for bioglass surface modification: Optimization studies.

    PubMed

    Lopes, João Henrique; Fonseca, Emanuella Maria Barreto; Mazali, Italo O; Magalhães, Alviclér; Landers, Richard; Bertran, Celso Aparecido

    2017-03-01

    In this work it is presented a facile and novel method for modification of bioglass surface based on (Camolten salt bath(2+)|Naglass(+)) ion exchange by immersion in molten salt bath. This method allows changing selectively the chemical composition of a surface layer of glass, creating a new and more reactive bioglass in a shell that surrounds the unchanged bulk of the original BG45S5 bioglass (core-shell type system). The modified bioglass conserves the non-crystalline structure of BG45S5 bioglass and presents a significant increase of surface reactivity in comparison with BG45S5. Melt-derived bioactive glasses BG45S5 with the nominal composition of 46.1mol% SiO2, 24.4mol% Na2O, 26.9mol% CaO, and 2.6mol% P2O5 have been subjected to ion exchange at 480°C in molten mixture of Ca(NO3)2 and NaNO3 with molar ratio of 70:30 for different time periods ranging from 0 to 60min. The optimization studies by using XRF and XRD showed that ion exchange time of 30min is enough to achieve higher changes on the glass surface without alters its non-crystalline structure. The chemical composition, morphology and structure of BG45S5 and bioglass with modified surface were studied by using several analytical techniques. FTIR and O1s XPS results showed that the modification of glass surface favors the formation of Si-ONBO groups at the expense of SiOBOSi bonds. (29)Si MAS-NMR studies showed that the connectivity of SiQ(n) species decreases from cross-linked SiQ(3) units to chain-like SiQ(2) units and finally to depolymerized SiQ(1) and SiQ(0) units after ion exchange. This result is consistent with the chemical model based on the enrichment with calcium ions of the bioglass surface such that the excess of positive charges is balanced by depolymerization of silicate network. The pH changes in the early steps of reaction of bioactive glasses BG45S5 and BG45Ca30, in deionized water or solutions buffered with HEPES were investigated. BG45Ca30 bioactive glass exhibited a significant

  10. 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.

  11. 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.

  12. 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

  13. Developing Critical Understanding in HRM Students: Using Innovative Teaching Methods to Encourage Deep Approaches to Study

    ERIC Educational Resources Information Center

    Butler, Michael J. R.; Reddy, Peter

    2010-01-01

    Purpose: This paper aims to focus on developing critical understanding in human resource management (HRM) students in Aston Business School, UK. The paper reveals that innovative teaching methods encourage deep approaches to study, an indicator of students reaching their own understanding of material and ideas. This improves student employability…

  14. 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.

  15. Developing Critical Understanding in HRM Students: Using Innovative Teaching Methods to Encourage Deep Approaches to Study

    ERIC Educational Resources Information Center

    Butler, Michael J. R.; Reddy, Peter

    2010-01-01

    Purpose: This paper aims to focus on developing critical understanding in human resource management (HRM) students in Aston Business School, UK. The paper reveals that innovative teaching methods encourage deep approaches to study, an indicator of students reaching their own understanding of material and ideas. This improves student employability…

  16. 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…

  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. 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…

  19. 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…

  20. Innovative methods for soil DNA purification tested in soils with widely differing characteristics.

    PubMed

    Sagova-Mareckova, Marketa; Cermak, Ladislav; Novotna, Jitka; Plhackova, Kamila; Forstova, Jana; Kopecky, Jan

    2008-05-01

    Seven methods of soil DNA extraction and purification were tested in a set of 14 soils differing in bedrock, texture, pH, salinity, moisture, organic matter content, and vegetation cover. The methods introduced in this study included pretreatment of soil with CaCO(3) or purification of extracted DNA by CaCl(2). The performance of innovated methods was compared to that of the commercial kit Mo Bio PowerSoil and the phenol-chloroform-based method of D. N. Miller, J. E. Bryant, E. L. Madsen, and W. C. Ghiorse (Appl. Environ. Microbiol. 65:4715-4724, 1999). This study demonstrated significant differences between the tested methods in terms of DNA yield, PCR performance, and recovered bacterial diversity. The differences in DNA yields were correlated to vegetation cover, soil pH, and clay content. The differences in PCR performances were correlated to vegetation cover and soil pH. The innovative methods improved PCR performance in our set of soils, in particular for forest acidic soils. PCR was successful in 95% of cases by the method using CaCl(2) purification and in 93% of cases by the method based on CaCO(3) pretreatment, but only in 79% by Mo Bio PowerSoil, for our range of soils. Also, the innovative methods recovered a higher percentage of actinomycete diversity from a subset of three soils. Recommendations include the assessment of soil characteristics prior to selecting the optimal protocol for soil DNA extraction and purification.

  1. [Maintenance of a male and a female clone of Schistosoma mansoni by microsurgical transplantation of sporocysts. Reliability of the method].

    PubMed

    Jourdane, J

    1984-01-01

    The long term maintenance (2 years) of a male clone and a female clone of Schistosoma mansoni by microsurgical transplantation of sporocysts allowed us to show that this technique is of great reliability: the postoperative survival of recipient snails at 60 days is greater than 75%; the mean of infection rate of the snails is 75% by the male clone sporocysts and 45% by the female clone sporocysts; the cercarial production of recipient snails is on average 441 cercariae per snail per day for the male clone and 530 cercariae for the female clone; the average worm recovery rate is 24,5%.

  2. Repeatable Construction Method for Engineered Zinc Finger Nuclease Based on Overlap Extension PCR and TA-Cloning

    PubMed Central

    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. PMID:23536890

  3. A novel sandwich hybridization method for selecting cDNAs from large genomic regions: Identification of cDNAs from the cloned genomic DNA spanning the XLRP locus

    SciTech Connect

    Yan, D.; McHenry, C.; Fujita, R.

    1994-09-01

    We have developed an efficient hybridization-based cDNA-selection method. A sandwich of three species - single-stranded cDNA, tagged RNA derived from genomic DNA, and biotinylated RNA complementary to the tag - allows specific retention of hybrids on an avidin-matrix. Previously, using model experiments, we demonstrated highly specific and efficient selection of a retinal gene, NRL, from complex mixtures of cDNA clones, using a sub-library from a 5 kb NRL genomic clone. We have now applied this selection strategy to isolate cDNAs from human adult retina and fetal eye libraries, with the {open_quotes}genomic RNA{close_quotes} derived from two YAC clones (OTC-C and 55B) spanning the region of X-linked retinitis pigmentosa (XLRP) locus RP3 at Xp21.1. Effectiveness of the selection-method was monitored by enrichment of TCTEX-1L gene that maps within the 55B YAC. Of the 15 selected cDNA clones that hybridized to the 55B YAC DNA, five appear to the map to specific cosmid clones derived from the 55B YAC. Inserts in these selected cDNA clones range from 0.5 to 2.3 kb in size. Additional clones are now being isolated and characterized. This procedure should be independent of the size or complexity of genomic DNA being used for selection, allow for the isolation of full-length cDNAs, and may have wider application.

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

    PubMed

    Stone, Vathsala I; Lane, Joseph P

    2012-05-16

    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. 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. 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 "bench to bedside" expectations for

  5. 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.

  6. 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.

  7. [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.

  8. [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.

  9. An Innovative Teaching Method To Promote Active Learning: Team-Based Learning

    NASA Astrophysics Data System (ADS)

    Balasubramanian, R.

    2007-12-01

    Traditional teaching practice based on the textbook-whiteboard- lecture-homework-test paradigm is not very effective in helping students with diverse academic backgrounds achieve higher-order critical thinking skills such as analysis, synthesis, and evaluation. Consequently, there is a critical need for developing a new pedagogical approach to create a collaborative and interactive learning environment in which students with complementary academic backgrounds and learning skills can work together to enhance their learning outcomes. In this presentation, I will discuss an innovative teaching method ('Team-Based Learning (TBL)") which I recently developed at National University of Singapore to promote active learning among students in the environmental engineering program with learning abilities. I implemented this new educational activity in a graduate course. Student feedback indicates that this pedagogical approach is appealing to most students, and promotes active & interactive learning in class. Data will be presented to show that the innovative teaching method has contributed to improved student learning and achievement.

  10. A Polymerase Chain Reaction-Based Method for Isolating Clones from a Complimentary DNA Library in Sheep

    PubMed Central

    Friis, Thor Einar; Stephenson, Sally; Xiao, Yin; Whitehead, Jon

    2014-01-01

    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. PMID:24447069

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

    DOE PAGES

    Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; ...

    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

  12. An efficient and high fidelity method for amplification, cloning and sequencing of complete tospovirus genomic RNA segments.

    PubMed

    Marshall, Spencer H; Adegbola, Raphael O; Adkins, Scott; Naidu, Rayapati A

    2017-04-01

    Tospoviruses (genus Tospovirus, family Bunyaviridae) are responsible for major losses in an extensive range of crops worldwide. New species of these single-stranded, ambisense RNA viruses regularly emerge and have been shown to maintain heterogeneous populations with individual isolates having quite variable biological and virulence characteristics. Most tospovirus phylogenetic studies have focused on analysis of a single gene, most often the nucleocapsid protein gene. Complete genomic RNA segment amplification as a single fragment would facilitate more detailed analyses of genome-wide sequence variability, but obtaining such sequences for a large number of tospovirus isolates using traditional methods of amplification and cloning of small overlapping fragments is tedious, time consuming and expensive. In this study, protocols were optimized to amplify, clone and sequence full-length M- and S-RNA genome segments of Tomato spotted wilt virus and Impatiens necrotic spot virus. The strategy presented here is straightforward, scalable and offers several advantages over the previously commonplace and overlapping amplicon-based approach. Use of whole genome segments, instead of individual gene sequences or defined portions of genome segments, will facilitate a better understanding of the underlying molecular diversity of tospoviruses in mixed infections.

  13. Rapid and efficient method for cloning of blunt-ended DNA fragments.

    PubMed

    Upcroft, P; Healey, A

    1987-01-01

    We describe a system to generate cDNA or genomic libraries from DNA segments that have blunt termini. Background and rearrangement levels are low, but efficiencies are high and the procedural times very short. T4 ligase in the presence of polyethylene glycol produces high Mr oligomers of vector and insert. These concatemers are reduced to vector-insert monomers at a high frequency by subsequent cleavage with a restriction endonuclease, which recognises the insert rarely, if at all, and the vector once. The monomers are recircularised under standard ligation conditions prior to transformation. Thus insertion conditions are optimised independently of those for recircularisation. All reading frames for expression libraries are generated by short BAL 31 cleavage followed by the blunt-end cloning procedure. Similarly, genomic expression libraries can be made by BAL 31 or mung-bean nuclease treatment after cleavage with DNase I is the presence of Mn2+. The technique is suitable for any DNA segment that is blunt-ended or can be made so. When the vector is treated with alkaline phosphatase, recombinants are generated at a frequency greater than 90% and have single inserts. Yields are 3-5 X 10(6) colony-forming units per micrograms of insert.

  14. Methods for Practising Ethics in Research and Innovation: A Literature Review, Critical Analysis and Recommendations.

    PubMed

    Reijers, Wessel; Wright, David; Brey, Philip; Weber, Karsten; Rodrigues, Rowena; O'Sullivan, Declan; Gordijn, Bert

    2017-09-12

    This paper provides a systematic literature review, analysis and discussion of methods that are proposed to practise ethics in research and innovation (R&I). Ethical considerations concerning the impacts of R&I are increasingly important, due to the quickening pace of technological innovation and the ubiquitous use of the outcomes of R&I processes in society. For this reason, several methods for practising ethics have been developed in different fields of R&I. The paper first of all presents a systematic search of academic sources that present and discuss such methods. Secondly, it provides a categorisation of these methods according to three main kinds: (1) ex ante methods, dealing with emerging technologies, (2) intra methods, dealing with technology design, and (3) ex post methods, dealing with ethical analysis of existing technologies. Thirdly, it discusses the methods by considering problems in the way they deal with the uncertainty of technological change, ethical technology design, the identification, analysis and resolving of ethical impacts of technologies and stakeholder participation. The results and discussion of our literature review are valuable for gaining an overview of the state of the art and serve as an outline of a future research agenda of methods for practising ethics in R&I.

  15. Effectiveness of various innovative learning methods in health science classrooms: a meta-analysis.

    PubMed

    Kalaian, Sema A; Kasim, Rafa M

    2017-01-13

    This study reports the results of a meta-analysis of the available literature on the effectiveness of various forms of innovative small-group learning methods on student achievement in undergraduate college health science classrooms. The results of the analysis revealed that most of the primary studies supported the effectiveness of the small-group learning methods in improving students' academic achievement with an overall weighted average effect-size of 0.59 in standard deviation units favoring small-group learning methods. The subgroup analysis showed that the various forms of innovative and reform-based small-group learning interventions appeared to be significantly more effective for students in higher levels of college classes (sophomore, junior, and senior levels), students in other countries (non-U.S.) worldwide, students in groups of four or less, and students who choose their own group. The random-effects meta-regression results revealed that the effect sizes were influenced significantly by the instructional duration of the primary studies. This means that studies with longer hours of instruction yielded higher effect sizes and on average every 1 h increase in instruction, the predicted increase in effect size was 0.009 standard deviation units, which is considered as a small effect. These results may help health science and nursing educators by providing guidance in identifying the conditions under which various forms of innovative small-group learning pedagogies are collectively more effective than the traditional lecture-based teaching instruction.

  16. The Business Engineering Surgical Technologies (BEST) teaching method: incubating talents for surgical innovation.

    PubMed

    de Ruijter, V; Halvax, P; Dallemagne, B; Swanström, L; Marescaux, J; Perretta, S

    2015-01-01

    Technological innovation in surgical science and healthcare is vital and calls for close collaboration between engineering and surgery. To meet this objective, BEST was designed as a free sustainable innovative teaching method for young professionals, combining surgery, engineering, and business in a multidisciplinary, high-quality, low-cost, and learning-by-doing philosophy. This paper reviews the initial outcomes of the program and discusses lessons learned and future directions of this innovative educational method. BEST educational method is delivered in two parts: the first component consisting of live streaming or pre-recorded online lectures, with an interdisciplinary profile focused on surgery, engineering, and business. The second component is an annual 5-day on-site course, organized at IRCAD-IHU, France. The program includes workshops in engineering, entrepreneurship team projects, and in-depth hands-on experience in laparoscopy, robotic surgery, interventional radiology, and flexible endoscopy with special emphasis on the interdisciplinary aspect of the training. A panel of surgeons, engineers, well-established entrepreneurs, and scientists assessed the team projects for potential patent application. From November 2011 till September 2013, 803 individual and institutional users from 79 different countries attended the online course. In total, 134 young professionals from 32 different countries applied to the onsite course. Sixty participants were selected each year for the onsite course. In addition, five participants were selected for a web-based team. Thirteen provisional patents were filed for the most promising projects. BEST proved to be a global talent incubator connecting students to high-quality education despite institutional and economical boundaries. Viable and innovative ideas arose from this revolutionary approach which is likely to spin-off significant technology transfer and lead the way for future interdisciplinary hybrid surgical

  17. Human Cloning

    DTIC Science & Technology

    2006-07-20

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

  18. An A-T linker adapter polymerase chain reaction method for chromosome walking without restriction site cloning bias.

    PubMed

    Trinh, Quoclinh; Xu, Wentao; Shi, Hui; Luo, Yunbo; Huang, Kunlun

    2012-06-01

    A-T linker adapter polymerase chain reaction (PCR) was modified and employed for the isolation of genomic fragments adjacent to a known DNA sequence. The improvements in the method focus on two points. The first is the modification of the PO(4) and NH(2) groups in the adapter to inhibit the self-ligation of the adapter or the generation of nonspecific products. The second improvement is the use of the capacity of rTaq DNA polymerase to add an adenosine overhang at the 3' ends of digested DNA to suppress self-ligation in the digested DNA and simultaneously resolve restriction site clone bias. The combination of modifications in the adapter and in the digested DNA leads to T/A-specific ligation, which enhances the flexibility of this method and makes it feasible to use many different restriction enzymes with a single adapter. This novel A-T linker adapter PCR overcomes the inherent limitations of the original ligation-mediated PCR method such as low specificity and a lack of restriction enzyme choice. Moreover, this method also offers higher amplification efficiency, greater flexibility, and easier manipulation compared with other PCR methods for chromosome walking. Experimental results from 143 Arabidopsis mutants illustrate that this method is reliable and efficient in high-throughput experiments.

  19. An "instant gene bank" method for heterologous gene cloning: complementation of two Aspergillus nidulans mutants with Gaeumannomyces graminis DNA.

    PubMed

    Bowyer, P; Osbourn, A E; Daniels, M J

    1994-02-01

    We present a novel technique for gene cloning by complementation of mutations in Aspergillus nidulans with DNA from a heterologous organism, Gaeumannomyces graminis. This technique bypasses the time-consuming and difficult construction of gene libraries, making it both rapid and simple. The method relies on recombination between a fungal replicating vector pHELP1 and linear G. graminis genomic DNA during co-transformation. We were able to complement two out of seven A. nidulans mutants tested and to rescue transforming DNA from both in Escherichia coli. Complementation of the A. nidulans argB mutation resulted from integration of 8-10 kb segments of G. graminis DNA into pHELP1. The complementation of the A. nidulans pyrG mutation resulted from a complex rearrangement. Complementing DNA was shown to originate from G. graminis, and was capable of retransforming the original mutants to give the expected phenotype.

  20. 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.

  1. Clone clustering by hybridization

    SciTech Connect

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

    1995-05-01

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

  2. Method-centered digital communities on protocols.io for fast-paced scientific innovation.

    PubMed

    Kindler, Lori; Stoliartchouk, Alexei; Teytelman, Leonid; Hurwitz, Bonnie L

    2016-01-01

    The Internet has enabled online social interaction for scientists beyond physical meetings and conferences. Yet despite these innovations in communication, dissemination of methods is often relegated to just academic publishing. Further, these methods remain static, with subsequent advances published elsewhere and unlinked. For communities undergoing fast-paced innovation, researchers need new capabilities to share, obtain feedback, and publish methods at the forefront of scientific development. For example, a renaissance in virology is now underway given the new metagenomic methods to sequence viral DNA directly from an environment. Metagenomics makes it possible to "see" natural viral communities that could not be previously studied through culturing methods. Yet, the knowledge of specialized techniques for the production and analysis of viral metagenomes remains in a subset of labs.  This problem is common to any community using and developing emerging technologies and techniques. We developed new capabilities to create virtual communities in protocols.io, an open access platform, for disseminating protocols and knowledge at the forefront of scientific development. To demonstrate these capabilities, we present a virology community forum called VERVENet. These new features allow virology researchers to share protocols and their annotations and optimizations, connect with the broader virtual community to share knowledge, job postings, conference announcements through a common online forum, and discover the current literature through personalized recommendations to promote discussion of cutting edge research. Virtual communities in protocols.io enhance a researcher's ability to: discuss and share protocols, connect with fellow community members, and learn about new and innovative research in the field.  The web-based software for developing virtual communities is free to use on protocols.io. Data are available through public APIs at protocols.io.

  3. Method-centered digital communities on protocols.io for fast-paced scientific innovation

    PubMed Central

    Kindler, Lori; Stoliartchouk, Alexei; Teytelman, Leonid; Hurwitz, Bonnie L.

    2017-01-01

    The Internet has enabled online social interaction for scientists beyond physical meetings and conferences. Yet despite these innovations in communication, dissemination of methods is often relegated to just academic publishing. Further, these methods remain static, with subsequent advances published elsewhere and unlinked. For communities undergoing fast-paced innovation, researchers need new capabilities to share, obtain feedback, and publish methods at the forefront of scientific development. For example, a renaissance in virology is now underway given the new metagenomic methods to sequence viral DNA directly from an environment. Metagenomics makes it possible to “see” natural viral communities that could not be previously studied through culturing methods. Yet, the knowledge of specialized techniques for the production and analysis of viral metagenomes remains in a subset of labs.  This problem is common to any community using and developing emerging technologies and techniques. We developed new capabilities to create virtual communities in protocols.io, an open access platform, for disseminating protocols and knowledge at the forefront of scientific development. To demonstrate these capabilities, we present a virology community forum called VERVENet. These new features allow virology researchers to share protocols and their annotations and optimizations, connect with the broader virtual community to share knowledge, job postings, conference announcements through a common online forum, and discover the current literature through personalized recommendations to promote discussion of cutting edge research. Virtual communities in protocols.io enhance a researcher’s ability to: discuss and share protocols, connect with fellow community members, and learn about new and innovative research in the field.  The web-based software for developing virtual communities is free to use on protocols.io. Data are available through public APIs at protocols.io. PMID

  4. 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…

  5. 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…

  6. 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

  7. 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.

  8. 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

  9. Innovation of floating time domain electromagnetic method in the case of environmental geophysics

    NASA Astrophysics Data System (ADS)

    Nurjanah, Siti; Widodo

    2017-07-01

    Geophysics has some methods that can be used to reveal the subsurface structure of the earth. The physical features obtained from the acquisition then analyzed and interpreted, so that it can be a great lead to interpret the physical contents, determine its position and its distribution. Geophysical methods also can be used to help the environment contamination survey which is referred to environmental geophysics. There are many sources of pollution that can harm the nature, for example, the source in the form of solid waste, liquid waste containing heavy metals, or radioactive, and etc. As time passes, these sources might settle in any sedimentary area and become sediments. Time Domain Electromagnetic (TDEM) is a trustworthy method to detect the presence of conductive anomaly due to sediment accumulation. Innovation of floating TDEM created to maximize the potential of the method, so that it can be used in aquatic environments. The configuration of TDEM modified using pipes and tires during the process of measurements. We conducted numerical simulation using Marquardt and Occam Algorithms towards synthetic model to ensure the capability of the proposed design. The development of this innovation is expected to be very useful to repair the natural conditions, especially in the water.

  10. Effect of roscovitine treated donor cells and different activation methods on development of handmade cloned goat (Capra hircus) embryos.

    PubMed

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

    2011-05-01

    The aim of the present investigation was to find out the effects of roscovitine treatment of donor cells and different activation methods on development of HMC goat embryos. Goat fetal fibroblast cells were cultured and divided into three treatment groups-contact inhibition group, roscovitine treatment group and serum starvation group. There was a significant decrease in blastocyst yield in serum starvation group (6.82%) compared to roscovitine treatment group (19.31%) and contact inhibition group (18.52%), however, no significant difference was found between roscovitine treatment group and contact inhibition group. To see the effect of different methods of activation, the reconstructed embryos were randomly divided into two groups and activated by two methods-one half by 2 μM Ca ionophore and another half by 2.31 kV/cm for 15 μSec electrical pulse. Subsequently, cloned embryos were cultured in TCM-199 based embryo development medium supplemented with 10 mg/mL bovine serum albumin in WOW culture system. There was a significant increase in the rate of cleavage and blastocyst production in electric pulse activation of 78.57% and 21.43% than Ca ionophore activation of 62.63% and 10.61% respectively. In conclusion, treatment of donor cells with roscovitine yields a significantly increased blastocyst than serum starved donor cells but equivalent blastocyst to contact inhibition group and electrical pulse activation (EPA) improves the production of HMC goat embryos. Copyright © 2011 Elsevier Inc. All rights reserved.

  11. Method to screen for relevant yeast two-hybrid-derived clones by coimmunoprecipitation and colocalization of epitope-tagged fragments--application to Bcl-xL.

    PubMed

    Wong, C; Naumovski, L

    1997-10-01

    The yeast two-hybrid system is a powerful genetic method to screen cDNA libraries to identify protein-protein interactions. A screen with a typical bait may yield many clones, including ones that are not biologically relevant which need to be eliminated by evaluating each clone in an alternative assay to confirm the interaction with the bait protein. We have developed an efficient assay to quickly screen two-hybrid-derived clones for coimmunoprecipitation and colocalization with the bait protein in mammalian cells. Gene fragments derived from a two-hybrid screen are cloned into an epitope tagging vector that can generate high levels of epitope-tagged protein in mammalian cells. The vector expressing an epitope-tagged protein is then cotransfected into mammalian cells with an expression vector for the bait protein. Interaction between the bait protein and epitope-tagged protein is evaluated by coimmunoprecipitation and colocalization. We demonstrate the utility of this approach by applying it to clones isolated in a two-hybrid screen using Bcl-xL as bait, showing that two-hybrid-derived fragments of Bad and Bax, previously known to interact with Bcl-xL, both colocalize and coimmunoprecipitate with Bcl-xL.

  12. [Human cloning or cannibalism].

    PubMed

    Sokolowski, L M

    2001-01-01

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

  13. Methods for assessment of innovative medical technologies during early stages of development.

    PubMed

    Bartelmes, Marc; Neumann, Ulrike; Lühmann, Dagmar; Schönermark, Matthias P; Hagen, Anja

    2009-11-05

    Conventional Health Technology Assessment (HTA) is usually conducted at a point in time at which the development of the respective technology may no longer be influenced. By this time developers and/or purchasers may have misinvested resources. Thus the demand for Technology Assessment (TA) which incorporates appropriate methods during early development stages of a technology becomes apparent. Against this health political background, the present report describes methods for a development-accompanying assessment of innovative medical technologies. Furthermore, international research programmes set out to identify or apply such methods will be outlined. A systematic literature search as well as an extensive manual literature search are carried out in order to obtain literature and information. The greatest units of the identified methods consist of assessment concepts, decision support methods, modelling approaches and methods focusing on users and their knowledge. Additionally, several general-purpose concepts have been identified. The identified research programmes INNO-HTA and MATCH (Multidisciplinary-Assessment-of-Technology-Centre-for-Healthcare) are to be seen as pilot projects which so far have not been able to generate final results. MATCH focuses almost entirely on the incorporation of the user-perspective regarding the development of non-pharmaceutical technologies, whereas INNO-HTA is basically concerned with the identification and possible advancement of methods for the early, socially-oriented technology assessment. Most references offer only very vague descriptions of the respective method and the application of greatly differing methods seldom exceeds the character of a pilot implementation. A standardisation much less an institutionalisation of development-accompanying assessment cannot be recognized. It must be noted that there is no singular method with which development-accompanying assessment should be carried out. Instead, a technology and

  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. Homopolymer tail-mediated ligation PCR: a streamlined and highly efficient method for DNA cloning and library construction

    PubMed Central

    Lazinski, David W.; Camilli, Andrew

    2013-01-01

    The amplification of DNA fragments, cloned between user-defined 5′ and 3′ end sequences, is a prerequisite step in the use of many current applications including massively parallel sequencing (MPS). Here we describe an improved method, called homopolymer tail-mediated ligation PCR (HTML-PCR), that requires very little starting template, minimal hands-on effort, is cost-effective, and is suited for use in high-throughput and robotic methodologies. HTML-PCR starts with the addition of homopolymer tails of controlled lengths to the 3′ termini of a double-stranded genomic template. The homopolymer tails enable the annealing-assisted ligation of a hybrid oligonucleotide to the template's recessed 5′ ends. The hybrid oligonucleotide has a user-defined sequence at its 5′ end. This primer, together with a second primer composed of a longer region complementary to the homopolymer tail and fused to a second 5′ user-defined sequence, are used in a PCR reaction to generate the final product. The user-defined sequences can be varied to enable compatibility with a wide variety of downstream applications. We demonstrate our new method by constructing MPS libraries starting from nanogram and sub-nanogram quantities of Vibrio cholerae and Streptococcus pneumoniae genomic DNA. PMID:23311318

  16. 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.

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

    PubMed

    Koniukhov, B V

    1997-12-01

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

  18. Why Clone?

    MedlinePlus

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

  19. A mixed methods study of emotional exhaustion: Energizing and depleting work within an innovative healthcare team.

    PubMed

    Cain, Cindy L; Taborda-Whitt, Caitlin; Frazer, Monica; Schellinger, Sandra; White, Katie M; Kaasovic, Jason; Nelson, Brenda; Chant, Allison

    2017-09-18

    This mixed methods study documents emotional exhaustion experiences among care team members during the development of an innovative team approach for caring for adults with serious illness. A mixed methods study design was employed to examine depleting work experiences that may produce emotional exhaustion, and energizing aspects of the work that may increase meaningfulness of work, thus reducing emotional exhaustion. The population studied included team members involved in care for adults with serious illness (n = 18). Team members were surveyed quarterly over an 18-month period using the Maslach Burnout Inventory (MBI). The MBI measures burnout, defined as the inability to continue work because of the interactional toll of the work. Analyses of MBI data show that although overall levels of burnout are low, 89% of team members reported moderate/high levels of emotional exhaustion during at least one survey period. In order to understand the kinds of work experiences that may produce or ameliorate emotional exhaustion, qualitative interviews were also conducted with team members at the end of the 18-month period. Major qualitative findings indicate that disputes within the team, environmental pressures, and standardisation of meaningful work leave team members feeling depleted. Having authentic relationships with patients, working as a team, believing in the care model, and practicing autonomy and creativity help team members to restore their emotional energy. Supports for team members' well-being are critical for continued innovation. We conclude with recommendations for improving team members' well-being.

  20. A novel cloning template designing method by using an artificial bee colony algorithm for edge detection of CNN based imaging sensors.

    PubMed

    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.

  1. 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

  2. Application of the ligation-independent cloning (LIC) method for rapid construction of a minigenome rescue system for Newcastle disease virus VG/GA strain.

    PubMed

    Zhao, Wei; Hu, Haixia; Zsak, Laszlo; Yu, Qingzhong; Yang, Zengqi

    2013-11-01

    Newcastle disease virus (NDV) can cause serious diseases and substantial economic losses to the poultry industry. To gain a better understanding of NDV pathogenesis, several reverse genetics systems for different NDV strains have been established. However, the construction of infectious cDNA clone by conventional restriction digestion/ligation cloning methods is a time-consuming process and has many drawbacks by its nature. To address the problems, we employed a novel and robust ligation-independent cloning (LIC) method for efficient assembly of multiple DNA fragments. Using this method, we successfully generated a NDV minigenome construct within three weeks by assembling RT-PCR products of the VG/GA strain genomic termini and a cDNA coding for the green fluorescence protein (GFP), as a reporter, into a modified pBluescript vector. Co-transfection of the NDV minigenome with three supporting plasmids expressing the N, P, and L proteins into MVA-T7 infected HEp-2 cells and followed by infection with NDV VG/GA resulted in the minigenome replication, transcription, and packaging as evidenced by the reporter gene GFP expression. These results suggest that this LIC approach is a powerful tool for all sequence-independent DNA cloning and multi-DNA fragment assembly, which has a potential application for rapid development of gene therapy and recombinant vaccines. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. 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.

  4. Topical Negative Pressure on Burns: An Innovative Method for Wound Exudate Collection

    PubMed Central

    Baudoin, Julien; Jafari, Paris; Meuli, Joachim; Raffoul, Wassim

    2016-01-01

    Summary: Burn wound exudate is an important source of information on the wound-healing process and systemic improvement of burn patients. Identification of biomarkers for the prediagnosis of local or systemic complications in patients will have a great impact on adapting personalized procedures in burn treatment. No efficient exudate collection method exists that offers a direct and continuous collection over time. We developed an innovative system based on the negative pressure wound therapy technique to directly collect exudate from burn wounds over several days after burn. This method did not cause any complication or pain for patients, and positive influence on wound healing was seen. Exudate samples were further used in different projects for studying biochemical profile, trace element content, kinetics of bacterial growth, and cell cytotoxicity. PMID:27975022

  5. Innovative method using magnetic particle tracking to measure solids circulation in a spouted fluidized bed

    SciTech Connect

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

    2009-01-01

    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.

  6. 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.

  7. Excision of DNA segments introduced into cloning vectors by the poly(dA-dT) joining method.

    PubMed Central

    Goff, S P; Berg, P

    1978-01-01

    A method is described for excising cloned DNA segments that have been inserted into their vectors by poly(dA-dT) joins. The recombinant DNA is cleaved within the vector DNA portion by one or more restriction endonucleases to generate a linear DNA molecule with the insert DNA sequence flanked by the poly(dA-dT) joins. After denaturation, the single strands "snap back" because of the intrastrand poly(dA) and poly(dT) sequences to form circular structures with "tails" of vector DNA. The vector portion of the DNA is then digested by Escherichia coli exonuclease VII, while the insert portion remains resistant to attack. The resistant strands are annealed and purified by electrophoresis in agarose. The insert DNA segment free of contaminating vector sequences can be used as a hybridization probe and for insertion into a new vector since suitable cohesive termini are generated from the retained poly(dA) and poly(dT) tails by an appropriate exonuclease. Images PMID:347445

  8. 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

  9. 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

  10. 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.

  11. [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.

  12. Diffusion of Innovations model helps interpret the comparative uptake of two methodological innovations: co-authorship network analysis and recommendations for the integration of novel methods in practice.

    PubMed

    Cadarette, Suzanne M; Ban, Joann K; Consiglio, Giulia P; Black, Cody D; Dubins, David; Marin, Alexandra; Tadrous, Mina

    2017-04-01

    The objective of this study was to characterize the diffusion of methodological innovation. Comparative case study analysis of the diffusion of two methods that summarize confounder information into a single score: disease risk score (DRS) and high-dimensional propensity score (hdPS). We completed systematic searches to identify DRS and hdPS papers in the field of pharmacoepidemiology through to the end of 2013, plotted the number of papers and unique authors over time, and created sociograms and animations to visualize co-authorship networks. First and last author affiliations were used to ascribe institutional contributions to each paper and network. We identified 43 DRS papers by 153 authors since 1981, reflecting slow uptake during initial periods of uncertainty and broader diffusion since 2001 linked to early adopters from Vanderbilt. We identified 44 hdPS papers by 147 authors since 2009, reflecting rapid and integrated diffusion, likely facilitated by opinion leaders, early presentation at conferences, easily accessible statistical code, and improvement in funding. Most contributions (87% DRS, 96% hdPS) were from North America. When proposing new methods, authors are encouraged to consider innovation attributes and early evaluation to improve knowledge translation of their innovations for integration into practice, and we provide recommendations for consideration. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  13. 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.

  14. 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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. 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

    2016-09-09

    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.

  16. 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

  17. Guide to molecular cloning techniques

    SciTech Connect

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

    1987-01-01

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

  18. 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

  19. Inferring cell type innovations by phylogenetic methods-concepts, methods, and limitations.

    PubMed

    Kin, Koryu

    2015-12-01

    Multicellular organisms are composed of distinct cell types that have specific roles in the body. Each cell type is a product of two kinds of historical processes-development and evolution. Although the concept of a cell type is difficult to define, the cell type concept based on the idea of the core regulatory network (CRN), a gene regulatory network that determines the identity of a cell type, illustrates the essential aspects of the cell type concept. The first step toward elucidating cell type evolution is to reconstruct the evolutionary relationships of cell types, or the cell type tree. The sister cell type model assumes that a new cell type evolves through divergence from a multifunctional ancestral cell type, creating tree-like evolutionary relationships between cell types. The process of generating a cell type tree can also be understood as the sequential addition of a new branching point on an ancestral cell differentiation hierarchy in evolution. A cell type tree thus represents an intertwined history of cell type evolution and development. Cell type trees can be reconstructed from high-throughput sequencing data, and the reconstruction of a cell type tree leads to the discovery of genes that are functionally important for a cell type. Although many issues including the lack of cross-species comparisons and the lack of a proper model for cell type evolution remain, the study of the origin of a new cell type using phylogenetic methods offers a promising new research avenue in developmental evolution. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 653-661, 2015. © 2015 Wiley Periodicals, Inc.

  20. Molecular cloning.

    PubMed

    Lessard, Juliane C

    2013-01-01

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

  1. Learning Evaluation: blending quality improvement and implementation research methods to study healthcare innovations.

    PubMed

    Balasubramanian, Bijal A; Cohen, Deborah J; Davis, Melinda M; Gunn, Rose; Dickinson, L Miriam; Miller, William L; Crabtree, Benjamin F; Stange, Kurt C

    2015-03-10

    In healthcare change interventions, on-the-ground learning about the implementation process is often lost because of a primary focus on outcome improvements. This paper describes the Learning Evaluation, a methodological approach that blends quality improvement and implementation research methods to study healthcare innovations. Learning Evaluation is an approach to multi-organization assessment. Qualitative and quantitative data are collected to conduct real-time assessment of implementation processes while also assessing changes in context, facilitating quality improvement using run charts and audit and feedback, and generating transportable lessons. Five principles are the foundation of this approach: (1) gather data to describe changes made by healthcare organizations and how changes are implemented; (2) collect process and outcome data relevant to healthcare organizations and to the research team; (3) assess multi-level contextual factors that affect implementation, process, outcome, and transportability; (4) assist healthcare organizations in using data for continuous quality improvement; and (5) operationalize common measurement strategies to generate transportable results. Learning Evaluation principles are applied across organizations by the following: (1) establishing a detailed understanding of the baseline implementation plan; (2) identifying target populations and tracking relevant process measures; (3) collecting and analyzing real-time quantitative and qualitative data on important contextual factors; (4) synthesizing data and emerging findings and sharing with stakeholders on an ongoing basis; and (5) harmonizing and fostering learning from process and outcome data. Application to a multi-site program focused on primary care and behavioral health integration shows the feasibility and utility of Learning Evaluation for generating real-time insights into evolving implementation processes. Learning Evaluation generates systematic and rigorous cross

  2. The model and the planning method of volume and variety assessment of innovative products in an industrial enterprise

    NASA Astrophysics Data System (ADS)

    Anisimov, V. G.; Anisimov, E. G.; Saurenko, T. N.; Sonkin, M. A.

    2017-01-01

    In the long term, the innovative development strategy efficiency is considered as the most crucial condition for assurance of economic system competitiveness in market conditions. It determines the problem relevance of such justification strategies with regard to specific systems features and conditions of their operation. The problem solution for industrial enterprises can be based on mathematical models of supporting the decision-making on the elements of the innovative manufacturing program. An optimization model and the planning method of innovative products volume and variety are suggested. The feature of the suggested model lies in the nonlinear nature of the objective function. It allows taking into consideration the law of diminishing marginal utility. The suggested method of optimization takes into account the system features and enables the effective implementation of manufacturing capabilities in modern conditions of production organization and sales in terms of market saturation.

  3. Alloreactive T cell clones.

    PubMed

    Fitch, F W

    1984-01-01

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

  4. An efficient method to enrich for knock-out and knock-in cellular clones using the CRISPR/Cas9 system.

    PubMed

    Niccheri, Francesca; Pecori, Riccardo; Conticello, Silvestro G

    2017-09-01

    Clustered Regularly Interspaced Short Palindromic Repeats-associated protein 9 nuclease (CRISPR/Cas9) and Transcription Activator-Like Effector Nucleases (TALENs) are versatile tools for genome editing. Here we report a method to increase the frequency of Cas9-targeted cellular clones. Our method is based on a chimeric construct with a Blasticidin S Resistance gene (bsr) placed out-of-frame by a surrogate target sequence. End joining of the CRISPR/Cas9-induced double-strand break on the surrogate target can place the bsr in frame, thus providing temporary resistance to Blasticidin S: this is used to enrich for cells where Cas9 is active. By this approach, in a real experimental setting, we disrupted the Aicda gene in ~70% of clones from CH12F3 lymphoma cells (>40% biallelically). With the same approach we knocked in a single nucleotide to reconstruct the frame of Aicda in these null cells, restoring the function in ~37% of the clones (less than 10% by the standard approach). Targeting of single nucleotide changes in other genes yielded analogous results. These results support our enrichment method as an efficient tool in genome editing.

  5. Challenges in Assessing Mobile Health App Quality: A Systematic Review of Prevalent and Innovative Methods.

    PubMed

    Grundy, Quinn H; Wang, Zhicheng; Bero, Lisa A

    2016-12-01

    Mobile apps are poised to become a major source of health guidance; however, concerns about their quality and safety remain. Governments, clinicians, and health researchers are struggling to determine how to adequately evaluate the content and function of mobile health apps to guide consumers toward apps that will effectively and safely support their health. Researchers are both adapting existing research methods, such as those for systematic reviews, and exploring novel methods toward this aim. The purpose of this systematic review was to describe prevalent and emerging methods for searching, data extraction, and analysis in the context of mobile health apps and commercial app stores. A systematic search was conducted on April 6, 2016, according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, for published studies that evaluated health-related apps targeted at consumers in a commercial app store, and a descriptive content analysis of their methods was performed. Although the majority of studies self-identified as "reviews" or "systematic reviews," their claims to have performed an exhaustive, replicable, and systematic search and data extraction are unsupported, given the nature of commercial app stores' search engines and personalized app content. Currently, studies of health-related app content are largely descriptive, and most use surrogate and one-dimensional outcome measures for app content quality. This field is rapidly developing, and innovative methods for systematic searches, multidimensional quality assessment, and contextualizing content assessment are emerging. Areas for future methodologic development are highlighted, including expanding definitions of app "content" and assessing risk of bias. Copyright © 2016 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

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

    NASA Astrophysics Data System (ADS)

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

    2011-02-01

    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 methods and to encourage students to continue in teacher education. We redesigned the elementary science methods course to include aspects of informal science education. The informal science education course features included informal science educator guest speakers, a live animal demonstration and a virtual field trip. We compared data from a treatment course ( n = 72) and a comparison course ( n = 26). Data collection included: researchers' observations, instructors' reflections, and teacher candidates' feedback. Teacher candidate feedback involved interviews and results on a reliable and valid Attitudes and Beliefs about the Nature of and the Teaching of Science instrument. We used complementary methods to analyze the data collected. A key finding of the study was that while benefits were found in both types of courses, the difference in results underscores the need of identifying the primary purpose for innovation as a vital component of consideration.

  7. Social networking and young adults' drinking practices: innovative qualitative methods for health behavior research.

    PubMed

    Lyons, Antonia C; Goodwin, Ian; McCreanor, Tim; Griffin, Christine

    2015-04-01

    Understandings of health behaviors can be enriched by using innovative qualitative research designs. We illustrate this with a project that used multiple qualitative methods to explore the confluence of young adults' drinking behaviors and social networking practices in Aotearoa, New Zealand. Participants were 18-25 year old males and females from diverse ethnic, class, and occupational backgrounds. In Stage 1, 34 friendship focus group discussions were video-recorded with 141 young adults who talked about their drinking and social networking practices. In Stage 2, 23 individual interviews were conducted using screen-capture software and video to record participants showing and discussing their Facebook pages. In Stage 3, a database of Web-based material regarding drinking and alcohol was developed and analyzed. In friendship group data, young adults co-constructed accounts of drinking practices and networking about drinking via Facebook as intensely social and pleasurable. However, this pleasure was less prominent in individual interviews, where there was greater explication of unpleasant or problematic experiences and practices. The pleasure derived from drinking and social networking practices was also differentiated by ethnicity, gender, and social class. Juxtaposing the Web-based data with participants' talk about their drinking and social media use showed the deep penetration of online alcohol marketing into young people's social worlds. Multiple qualitative methods, generating multimodal datasets, allowed valuable nuanced insights into young adults' drinking practices and social networking behaviors. This knowledge can usefully inform health policy, health promotion strategies, and targeted health interventions. (c) 2015 APA, all rights reserved).

  8. 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. © 2014 American Academy of Psychiatry and the Law.

  9. Microelectro discharge machining: an innovative method for the fabrication of 3D microdevices

    NASA Astrophysics Data System (ADS)

    Lesche, Claudia; Krah, Thomas; Büttgenbach, Stephanus

    2011-06-01

    This paper reports on the potential of microelectro discharge machining (μEDM) as an innovative method for the fabrication of 3D microdevices. To demonstrate the wide capabilities of μEDM two different high-potential 3D microsystems - a microfluidic device for the dispersion of nanoparticles and a star probe for microcoordinate metrology - are presented. For the fabrication of these microdevices a μEDM-milling machine with integrated microwire electro discharge grinding (μWEDG) module is utilized. To gain optimized process conditions as well as a high surface quality an adequate adaption of the single erosion parameters such as energy, pulse frequency and spark gap has to be carried out and are discussed below. The dispersion micromodule is used for pharmaceutical screening applications in a high pressure range up to 2000 bar. At the channel bottom a surface roughness of Ra = 80 nm is achieved. In case of the star probe it is possible to produce shaft and sphere out of one piece. The fabricated stylus elements have sphere diameters of 40-200 μm. For both applications μEDM offers a flexible, precise, effective and cost-efficient fabrication method for the machining of hard and resistant materials.

  10. An Innovative Method for Evaluating Strategic Goals in a Public Agency: Conservation Leadership in the U.S. Forest Service.

    ERIC Educational Resources Information Center

    Bengston, David N.; Fan, David P.

    1999-01-01

    Presents an innovative methodology for evaluating strategic goals in a public agency. The method involves computer content analysis of online news media text to evaluate expressed attitudes. Provides a way to assess the views of a wide range of stakeholders quickly and efficiently. (SLD)

  11. Potential for cloning dogs.

    PubMed

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

    2001-01-01

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

  12. A new innovative spectrophotometric method for the simultaneous determination of sofosbuvir and ledipasvir.

    PubMed

    Mansour, Fotouh R

    2017-07-31

    A new innovative spectrophotometric method is developed to determine the concentration ratios in binary mixtures by determining the zero crossing point in the first derivative of the gross curve. This relationship can be applied if the part of the UV spectrum of substance Y, that intersects with the overlaid spectra of substance X is straight. By plotting the intersection wavelength against the concentration ratio (CxCy), a straight line was obtained with a co-efficient of determination equals 0.9999. As an application, simultaneous determination of sofosbuvir and ledipasvir in their binary mixtures was performed using two methods; a direct UV spectrophotometric method for determination of ledipasvir at 333nm, and the new "wavelength-intersection ratio" method for determination of sofosbuvir. In the wavelength-intersection ratio method, different mixtures of sofosbuvir and ledipasvir containing different concentration ratios were prepared; the zero crossing point of the first derivative curve in the range 285 to 295nm were determined for each mixture. An absorbance shift in the intersection was obtained with the change in the concentration ratio (sofosbuvir/ledipasvir). When the concentration ratio was plotted against the intersection wavelength, a straight line was obtained with a coefficient of determination of 0.9992. The direct method was linear in the range 3 to 18μg/mL while the wavelength-intersection ratio method was linear in the range 11-110μg/mL. The limits of detection were determined and found to be 0.5 and 3μg/mL for ledipasvir and sofosbuvir, respectively. The accuracy and repeatability of the two methods were tested. The mean %recovery was found to be 100.19% and 100.75% for ledipasvir and sofosbuvir, respectively. The relative standard deviation was 0.57 for ledipasvir and 1.79 for sofosbuvir. The intermediate precision was also checked, the coefficients of variation for sofosbuvir and ledipasvir measurements between days did not exceed 1

  13. Development of large DNA methods for plants: molecular cloning of large segments of Arabidopsis and carrot DNA into yeast.

    PubMed Central

    Guzmán, P; Ecker, J R

    1988-01-01

    Procedures for the preparation, analysis and cloning of large DNA molecules from two different plant species are described. Arabidopsis and carrot protoplasts were used for the preparation of large DNA molecules in agarose "plugs" or in solution. Pulsed-field gel electrophoresis (PFGE) analysis of large plant DNA preparations using a contour-clamped homogeneous field (CHEF) apparatus indicated that the size of the DNA was at least 12 Mb. Large DNA preparations were shown to be useful for restriction enzyme analysis of the Arabidopsis genome using both frequent and infrequent cutting enzymes and for the molecular cloning of large segments of DNA into yeast using artificial chromosome (YAC) vectors. PFGE and blot hybridization analysis of Arabidopsis and carrot DNA-containing YACs indicated that both unique and highly repeated DNA sequences were represented in these libraries. Images PMID:3060856

  14. Using qualitative research methods in biomedical innovation: the case of cultured red blood cells for transfusion.

    PubMed

    Lyall, Catherine; King, Emma

    2016-05-11

    Qualitative research has a key role to play in biomedical innovation projects. This article focuses on the appropriate use of robust social science methodologies (primarily focus group studies) for identifying the public's willingness and preference for emerging medical technologies. Our study was part of the BloodPharma project (now known as the Novosang project) to deliver industrially generated red blood cells for transfusion. Previous work on blood substitutes shows that the public prefers donated human blood. However, no research has been conducted concerning attitudes to stem cell derived red blood cells. Qualitative research methods including interviews and focus groups provide the methodological context for this paper. Focus groups were used to elicit views from sub-sections of the UK population about the potential use of such cultured red blood cells. We reflect on the appropriateness of that methodology in the context of the BloodPharma project. Findings are in the form of lessons transferable to other interdisciplinary, science-led teams about what a social science dimension can bring; why qualitative research should be included; and how it can be used effectively. Qualitative data collection offers the strength of exploring ambivalence and investigating the reasons for views, but not necessarily their prevalence in wider society. The inherent value of a qualitative method, such as focus groups, therefore lies in its ability to uncover new information. This contrasts with a quantitative approach to simply 'measuring' public opinion on a topic about which participants may have little prior knowledge. We discuss a number of challenges including: appropriate roles for embedded social scientists and the intricacies of doing upstream engagement as well as some of the design issues and limitations associated with the focus group method.

  15. 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.

  16. 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.

  17. 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...

  18. Secure the Clones

    NASA Astrophysics Data System (ADS)

    Jensen, Thomas; Kirchner, Florent; Pichardie, David

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

  19. Molecular cloning of Indian tomato leaf curl virus genome following a simple method of concentrating the supercoiled replicative form of viral DNA.

    PubMed

    Srivastava, K M; Hallan, V; Raizada, R K; Chandra, G; Singh, B P; Sane, P V

    1995-02-01

    DNA-A and DNA-B components of the genome of a whitefly transmitted virus causing yellowing and leaf curl in tomato (ITLCV) were cloned following a simple procedure for isolation of the double stranded replicative form of viral DNA from infected tomato plants. The method is based on extraction of total DNA from infected plants followed by concentration of the double stranded replicative form of viral DNA by an alkaline denaturation procedure identical to that used for isolation of plasmid DNA from Escherichia coli. The attempted cloning of DNA showed that 95% of the transformants contained plasmids with an insert of either DNA-A (2.75 kb) or DNA-B (2.55 kb). Cloned DNA-A and DNA-B when used as probes could detect DNA-A/DNA-B in total nucleic acid obtained from fresh diseased tissue. Both DNA-A and DNA-B are needed for infection and they have a common region of 166 bases with about 94% nucleotide sequence homology, a characteristic of all bipartite geminiviruses. Comparison of the amino acid sequence of the putative coat protein product of ITLCV with some other mono- and bipartite geminiviruses revealed a maximum of 86% homology with Indian cassava mosaic virus.

  20. 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.

  1. 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

  2. Evaluation of Students' Perceptions Towards An Innovative Teaching-Learning Method During Pharmacology Revision Classes: Autobiography of Drugs.

    PubMed

    Joshi, Anuradha; Ganjiwale, Jaishree

    2015-07-01

    Various studies in medical education have shown that active learning strategies should be incorporated into the teaching-learning process to make learning more effective, efficient and meaningful. The aim of this study was to evaluate student's perceptions on an innovative revision method conducted in Pharmacology i.e. in form of Autobiography of Drugs. The main objective of study was to help students revise the core topics in Pharmacology in an interesting way. Questionnaire based survey on a newer method of pharmacology revision in two batches of second year MBBS students of a tertiary care teaching medical college. Various sessions on Autobiography of Drugs were conducted amongst two batches of second year MBBS students, during their Pharmacology revision classes. Student's perceptions were documented with the help of a five point likert scale through a questionnaire regarding quality, content and usefulness of this method. Descriptive analysis. Students of both the batches appreciated the innovative method taken up for revision. The median scores in most of the domains in both batches were four out of five, indicative of good response. Feedback from open-ended questions also revealed that the innovative module on "Autobiography of Drugs" was taken as a positive learning experience by students. Autobiography of drugs has been used to help students recall topics that they have learnt through other teachings methods. Autobiography sessions in Pharmacology during revision slots, can be one of the interesting ways in helping students revise and recall topics which have already been taught in theory classes.

  3. 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.

  4. Reduction of membrane fouling by innovative method (injection of air jet).

    PubMed

    Amiraftabi, Maryam-Sadat; Mostoufi, Navid; Hosseinzadeh, Mostafa; Mehrnia, Mohammad-Reza

    2014-01-01

    One of the most important challenges about the Membrane Bio Reactors is membrane fouling. Fouling has been at the centre of a globe debate for more recent years. It leads to high operational and maintenance costs such as membrane damage and replacement of membrane. Membrane fouling is attributed to the physicochemical interactions between the bio fluid and membrane. In order to decrease the fouling in bioreactors there are common anti fouling strategies such as operation at low flux, Optimization of aeration flow-rate and Physical and chemical cleanings. However, often they are not effective. This work deal with fouling crisis by a new and innovative method in order to reduce of fouling on membrane surface by injection of parallel air jet on membrane bio reactor. This is a new idea and fundamental study about the influence of wall jet on fouling of membrane surface. This study is included both experimental and numerical investigations. In order to polarize the stream path on the surface of the membrane, four symmetric nozzles were implemented at the bottom of the membrane surface upon the sparger. The changes in the fouling resistance were experimentally measured at five various jet velocities and all of them recorded by a computer system. In addition the effect of air jet velocity and shear stress on fouling resistances was also investigated by computational fluid dynamics at the similar conditions. It was revealed that the permeate flux and resistance of fouling can be related to shear stress of air flow at the membrane surface. When the velocity of air jets increase, the permeate flux increase too. Also, results illustrate that jet injection can partially remove the cake which was formed on the surface of the membrane. Correlations were developed for estimating each resistance of the membrane surface via the shear stress. The resistances of the cake are removed by the jet velocity changes, from 20% in lower jet velocity up to 40% in higher jet velocity.

  5. Making the cut: Innovative methods for optimizing perfusion-based migration assays.

    PubMed

    Holt, Andrew W; Howard, William E; Ables, Elizabeth T; George, Stephanie M; Kukoly, Cindy A; Rabidou, Jake E; Francisco, Jake T; Chukwu, Angel N; Tulis, David A

    2017-03-01

    Application of fluid shear stress to adherent cells dramatically influences their cytoskeletal makeup and differentially regulates their migratory phenotype. Because cytoskeletal rearrangements are necessary for cell motility and migration, preserving these adaptations under in vitro conditions and in the presence of fluid flow are physiologically essential. With this in mind, parallel plate flow chambers and microchannels are often used to conduct in vitro perfusion experiments. However, both of these systems currently lack capacity to accurately study cell migration in the same location where cells were perfused. The most common perfusion/migration assays involve cell perfusion followed by trypsinization which can compromise adaptive cytoskeletal geometry and lead to misleading phenotypic conclusions. The purpose of this study was to quantitatively highlight some limitations commonly found with currently used cell migration approaches and to introduce two new advances which use additive manufacturing (3D printing) or laser capture microdissection (LCM) technology. The residue-free 3D printed insert allows accurate cell seeding within defined areas, increases cell yield for downstream analyses, and more closely resembles the reported levels of fluid shear stress calculated with computational fluid dynamics as compared to other residue-free cell seeding techniques. The LCM approach uses an ultraviolet laser for "touchless technology" to rapidly and accurately introduce a custom-sized wound area in otherwise inaccessible perfusion microchannels. The wound area introduced by LCM elicits comparable migration characteristics compared to traditional pipette tip-induced injuries. When used in perfusion experiments, both of these newly characterized tools were effective in yielding similar results yet without the limitations of the traditional modalities. These innovative methods provide valuable tools for exploring mechanisms of clinically important aspects of cell

  6. [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.

  7. 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)

  8. 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)

  9. 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)

  10. [Eugenics and human cloning].

    PubMed

    Boloz, W

    2001-01-01

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

  11. Innovative Teaching Ideas. A Sharing of Teaching Tips, Ideas and Methods.

    ERIC Educational Resources Information Center

    Tenney, Richard W., Ed.; And Others

    Compiled from materials collected from 150 teachers of agriculture across New York State who participated in a series of 11 regional agriculture inservice workshops, this resource guide contains innovative techniques for classroom operation as well as suggestions for using available resources. The ideas have been organized into the following eight…

  12. 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.

  13. 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.

  14. 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…

  15. 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,…

  16. 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,…

  17. 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).

  18. 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. Copyright © 2011 Elsevier Inc

  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. Do International Health Partnerships contribute to reverse innovation? a mixed methods study of THET-supported partnerships in the UK.

    PubMed

    Kulasabanathan, Kavian; Issa, Hamdi; Bhatti, Yasser; Prime, Matthew; Del Castillo, Jacqueline; Darzi, Ara; Harris, Matthew

    2017-04-18

    International health partnerships (IHPs) are changing, with an increased emphasis on mutual accountability and joint agenda setting for both the high- and the low- or middle-income country (LMIC) partners. There is now an important focus on the bi-directionality of learning however for the UK partners, this typically focuses on learning at the individual level, through personal and professional development. We sought to evaluate whether this learning also takes the shape of 'Reverse Innovation' -when an idea conceived in a low-income country is subsequently adopted in a higher-income country. This mixed methods study used an initial scoping survey of all the UK-leads of the Tropical Health Education Trust (THET)-supported International Health Partnerships (n = 114) to ascertain the extent to which the IHPs are or have been vehicles for Reverse Innovation. The survey formed the sampling frame for further deep-dive interviews to focus on volunteers' experiences and attitudes to learning from LMICs. Interviews of IHP leads (n = 12) were audio-recorded and transcribed verbatim. Survey data was analysed descriptively. Interview transcripts were coded thematically, using an inductive approach. Survey response rate was 27% (n = 34). The majority (70%) strongly agreed that supporting LMIC partners best described the mission of the partnership but only 13% of respondents strongly agreed that learning about new innovations and models was a primary mission of their partnership. Although more than half of respondents reported having observed innovative practice in the LMIC, only one IHP respondent indicated that this has led to Reverse Innovation. Interviews with a sample of survey respondents revealed themes primarily around how learning is conceptualised, but also a central power imbalance between the UK and LMIC partners. Paternalistic notions of knowledge could be traced to partnership power dynamics and latent attitudes to LMICs. Given the global flow of

  1. Field analytical screening program: PCP method. Innovative technology evaluation report. Report for 1 July-31 August 1993

    SciTech Connect

    Hess, E.; Hamilton, D.

    1995-08-01

    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 pentachlorophenol (PCP) contamination in soil and water. This method was demonstrated in Norrisville, North Carolina, in August 1993. The FASP PCP Method was developed by the EPA Superfund Branch for use at Superfund sites. The method uses a gas chromatograph (GC) equipped with a megabore capillary column and either a flame ionization detector (FID) or an electron capture detector (ECD). Gas chromatography is an EPA-approved method for determining PCP concentrations in soil, water, and waste samples. The FASP PCP Method is an abbrevaited, modified version of approved methods. Soil and water samples require extraction before GC analysis. To remove interferences caused by petroleum hydrocarbons, an acid-base partition cleanup step is used during the FASP PCP Method.

  2. CRISPR/Cas9-mediated gene manipulation to create single-amino-acid-substituted and floxed mice with a cloning-free method.

    PubMed

    Ma, Xiaolong; Chen, Chao; Veevers, Jennifer; Zhou, XinMin; Ross, Robert S; Feng, Wei; Chen, Ju

    2017-02-08

    Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology is a powerful tool to manipulate the genome with extraordinary simplicity and speed. To generate genetically modified animals, CRISPR/Cas9-mediated genome editing is typically accomplished by microinjection of a mixture of Cas9 DNA/mRNA and single-guide RNA (sgRNA) into zygotes. However, sgRNAs used for this approach require manipulation via molecular cloning as well as in vitro transcription. Beyond these complexities, most mutants obtained with this traditional approach are genetically mosaic, yielding several types of cells with different genetic mutations. Recently, a growing body of studies has utilized commercially available Cas9 protein together with sgRNA and a targeting construct to introduce desired mutations. Here, we report a cloning-free method to target the mouse genome by pronuclear injection of a commercial Cas9 protein:crRNA:tracrRNA:single-strand oligodeoxynucleotide (ssODN) complex into mouse zygotes. As illustration of this method, we report the successful generation of global gene-knockout, single-amino-acid-substituted, as well as floxed mice that can be used for conditional gene-targeting. These models were produced with high efficiency to generate non-mosaic mutant mice with a high germline transmission rate.

  3. CRISPR/Cas9-mediated gene manipulation to create single-amino-acid-substituted and floxed mice with a cloning-free method

    PubMed Central

    Ma, Xiaolong; Chen, Chao; Veevers, Jennifer; Zhou, XinMin; Ross, Robert S.; Feng, Wei; Chen, Ju

    2017-01-01

    Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology is a powerful tool to manipulate the genome with extraordinary simplicity and speed. To generate genetically modified animals, CRISPR/Cas9-mediated genome editing is typically accomplished by microinjection of a mixture of Cas9 DNA/mRNA and single-guide RNA (sgRNA) into zygotes. However, sgRNAs used for this approach require manipulation via molecular cloning as well as in vitro transcription. Beyond these complexities, most mutants obtained with this traditional approach are genetically mosaic, yielding several types of cells with different genetic mutations. Recently, a growing body of studies has utilized commercially available Cas9 protein together with sgRNA and a targeting construct to introduce desired mutations. Here, we report a cloning-free method to target the mouse genome by pronuclear injection of a commercial Cas9 protein:crRNA:tracrRNA:single-strand oligodeoxynucleotide (ssODN) complex into mouse zygotes. As illustration of this method, we report the successful generation of global gene-knockout, single-amino-acid-substituted, as well as floxed mice that can be used for conditional gene-targeting. These models were produced with high efficiency to generate non-mosaic mutant mice with a high germline transmission rate. PMID:28176880

  4. Characterization of the regulatory region of the dopa decarboxylase gene in Medaka: an in vivo green fluorescent protein reporter assay combined with a simple TA-cloning method.

    PubMed

    Fujimori, Kazuhiro E

    2009-03-01

    The mechanism by which differentiated cells cooperatively express specific sets of genes in multicellular organisms is a fundamental question for biologists. Currently, the mechanism is primarily attributed to complex regulation of transcriptional machinery. Here, I provide a method for studying spatiotemporal characteristics of promoters in vivo by rapid construction of reporter gene-expression vectors based on simple TA-cloning using an in vivo eGFP reporter assay in Medaka (Oryzias latipes). As an application of this method, I focused on the dopa decarboxylase (Ddc) gene, an essential enzyme for production of neurotransmitters, dopamine, and serotonin. Based on the known structure of the Medaka genome, I predicted and cloned the approximately 3 kbp fragment flanking the Ddc gene. Using an eGFP reporter assay in vivo, I showed that it functions as a promoter, directing reporter gene expression in the brain, retina, epiphysis, and gut, but not in sympathetic ganglia, kidney, or liver. Thus, the procedure presented here provides a useful tool for rapid screening of possible promoter regions and for establishing germ line-transmitted transgenic lines of Medaka.

  5. 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

  6. Rehabilitation of gingival architecture by a conservative method: An innovative approach

    PubMed Central

    Parolia, Abhishek; Thomas, Manuel S; Kundabala, M; Shetty, Neeta; Gautam, Suman; Kumar, Senthil

    2008-01-01

    Periodontal attachment loss in the maxillary anterior region can often lead to esthetic and functional clinical problems. Lifelong motivation is essential to the supportive therapy for these patients, and the maintenance of good esthetics, combined with conducive to maintaining long term dental and professional health. This paper aims to demonstrate an innovative treatment option for dealing with aesthetic challenges posed by a number of patients who have undergone initial cause related therapy for aggressive periodontitis. PMID:20142901

  7. A method for identification of the peptides that bind to a clone of thyroid-stimulating antibodies in the serum of Graves' disease patients.

    PubMed

    Na, Chan Hyun; Lee, Mi Hwa; Cho, Bo Youn; Chae, Chi-Bom

    2003-04-01

    A method was developed for identification of the peptide sequences that bind to thyroid-stimulating antibody (TSAb) clones from phage-displayed peptide library. Immunoglobulin G (IgG) was purified from the serum of a Graves' disease patient that stimulates the synthesis of cAMP in the cells that express TSH receptor (TSHR). The IgG that binds to TSHR was purified by an affinity column packed with the resin cross-linked with the extracellular domain of human TSHR. The receptor-binding IgG was then mixed with phages that display linear or cyclic peptides at the end of tail protein pIII. The bound phages were eluted with acidic glycine after extensive washing. From sequencing of the pIII gene of the bound phages, one can deduce the sequences of the peptides that bind to the receptor-binding IgG. Each peptide sequence was then tested for inhibition of the synthesis of cAMP from thyroid cells induced by the serum of a Graves' patient. In this way, one can obtain the peptides that bind to a clone of TSAb. We obtained a peptide sequence that inhibits the action of TSAb at an extremely low concentration (<10(-14) M). Such a peptide will be useful for various studies on TSAb.

  8. 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.

  9. 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…

  10. 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…

  11. An Innovative Method for Low Cost, Autonomous Navigation for Low Earth Orbit Satellites

    NASA Technical Reports Server (NTRS)

    Deutschmann, Julie; Harman, Rick; Bar-Itzhack, Itzhack

    1998-01-01

    An innovative approach to autonomous attitude, trajectory, and rate estimation is presented for low earth orbit (LEO) satellites which relies on magnetometers and sun sensors. These two sensors are reliable, inexpensive, and are used routinely in LEO missions for attitude determination and control. An extended Kalman filter (EKF) is developed from two existing systems, one which uses an EKF to estimate attitude and trajectory using magnetometer and gyro data and a second pseudo-linear filter which estimates rotation rate using magnetometer and sun sensor data. The theoretical background of the combined system is presented along with test results from noisy, simulated sensor data.

  12. An innovative method of generating current and thermoelectric equipment for its realization

    NASA Astrophysics Data System (ADS)

    Kušnerová, M.; Valíček, J.; Harničárová, M.; Koštial, P.; Jančíková, Z.

    2015-02-01

    The paper assesses an innovative thermoelectric device used to generate a current by the conversion of thermal energy into electrical energy. The device has been created and verified. Until now, the efficiency of conventional thermoelectric batteries is for technical practice not sufficiently appreciated, because it does not exceed 3 %. A necessary condition needed for the implementation of the Seebeck thermoelectric effect is a sufficient and stable source of heat that provides an optimum temperature difference, but there are two other sufficient conditions for a significant increase in efficiency of thermoelectric batteries. These are concerned with the development of new materials for thermoelectric batteries and with the development of new construction of thermoelectric batteries.

  13. New methods for innovation: the development of a toolbox for natural orifice translumenal endoscopic surgery (NOTES) procedures.

    PubMed

    Swain, C Paul; Bally, Kurt; Park, Per-Ola; Mosse, C Alexander; Rothstein, Richard I

    2012-04-01

    Devices used for flexible intralumenal procedures are inadequate when used for intraperitoneal surgical procedures such as cholecystectomy. To assess/address limitations of flexible endoscopic devices in intraperitoneal surgery. To describe processes used to invent new devices to facilitate this new surgical genre. Engineering laboratory. None. INTERVENTIONS AND INVENTIONS: Reviews of the limitations of flexible endoscopic instruments and instrumentation/invention needs for a "NOTES cholecystectomy" were completed. The appropriateness of existing methods of device innovation was evaluated against an inventory of new technologies necessary to perform NOTES. The deficiencies in traditional innovation methods led to the creation of a novel process for invention of new medical devices: the "Inventorama." Cooperation between clinicians and industry to develop device concepts to enable NOTES. The devices included: (1) steerable flex trocar, (2) rotary access needle, (3) bipolar hemostasis forceps, (4) Maryland dissectors, (5) articulating hook knife, (6) rotating hook knife, (7) articulating graspers, (8) scissors, (9) ligating clip applier, and (10) tissue apposition system. Six of these ten were built and tested as initial crude prototypes in the Inventorama process; two underwent major modifications. Three were invented via alternate methods, including by independent clinicians. A new method for efficient medical device invention and development was created to address key technology needs for NOTES. The result was a "toolbox" of devices designed to address the key surgical activities necessary for advanced intralumenal and translumenal flexible endoscopic procedures.

  14. 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.

  15. Networked Innovation in Innovation Networks: A Home Appliances Case Study

    NASA Astrophysics Data System (ADS)

    Berasategi, Luis; Arana, Joseba; Castellano, Eduardo

    Amongst different types of Collaborative Networked Organizations it is possible to highlight those created to develop and market product, process or business model innovation. In this type of innovation network, which has special characteristics, the challenge is to introduce effective networked innovation in the very same innovation network. This paper presents the main features of TALAI-SAREA © methodology that includes a reference model, a set of analysis tools and a method for implementing networked innovation in innovation networks.

  16. 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.

  17. 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.

  18. 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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Innovative methods and tools for professionals working in supported living services for intellectually disabled persons.

    PubMed

    Gruiz, Katalin

    2015-01-01

    Autonomy of mid-seriously and seriously intellectually disabled persons is encouraged both by legislations on human rights and the modern social care and services. The process leading to the maximum possible autonomy is illustrated by a developmental spiral in our model. Specialty of the development is that the personal educational projects are realized during everyday activities. The process requires conscious professionals with an empowering and motivating attitude, with adult relationship to the intellectually disabled persons and versatile skills and tools. In this educational relationship the social professional and the supported person are equal partners moving together along the spiral of human development. An innovative tool-battery has been developed aiding support-staff in the 'pedagogical' task embedded into everyday social services. The tool-battery and its first application in supported living services of the Hungarian Down Foundation are introduced in this paper.

  20. 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

  1. 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.

  2. Innovation in Surgery

    PubMed Central

    Riskin, Daniel J.; Longaker, Michael T.; Gertner, Michael; Krummel, Thomas M.

    2006-01-01

    Objective: To describe the field of surgical innovation from a historical perspective, applying new findings from research in technology innovation. Background: While surgical innovation has a rich tradition, as a field of study it is embryonic. Only a handful of academic centers of surgical innovation exist, all of which have arisen within the last 5 years. To this point, the field has not been well defined, nor have future options to promote surgical innovation been thoroughly explored. It is clear that surgical innovation is fundamental to surgical progress and has significant health policy implications. A process of systematically evaluating and promoting innovation in surgery may be critical in the evolving practice of medicine. Methods: A review of the academic literature in technology innovation was undertaken. Articles and books were identified through technical, medical, and business sources. Luminaries in surgical innovation were interviewed to develop further relevance to surgical history. The concepts in technology innovation were then applied to innovation in surgery, using the historical example of surgical endoscopy as a representative area, which encompasses millennia of learning and spans multiple specialties of care. Results: The history of surgery is comprised largely of individual, widely respected surgeon innovators. While respecting individual accomplishments, surgeons as a group have at times hindered critical innovation to the detriment of our profession and patients. As a clinical discipline, surgery relies on a tradition of research and attracting the brightest young minds. Innovation in surgery to date has been impressive, but inconsistently supported. Conclusion: A body of knowledge on technology innovation has been developed over the last decade but has largely not been applied to surgery. New surgical innovation centers are working to define the field and identify critical aspects of surgical innovation promotion. It is our

  3. From deep sequencing to actual clones.

    PubMed

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

    2014-10-01

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

  4. 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.

  5. An efficient and rapid method for cDNA cloning from difficult templates using codon optimization and SOE-PCR: with human RANK and TIMP2 gene as examples.

    PubMed

    Huang, Gang; Wen, Qianjun; Gao, Qiangguo; Zhang, Fang; Bai, Yun

    2011-10-01

    As gene cloning from difficult templates with regionalized high GC content is a long recognized problem, we have developed a novel and reliable method to clone such genes. Firstly, the high GC content region of the target cDNA was synthesized directly after codon optimization and the remaining cDNA fragment without high GC content was generated by routine RT-PCR. Then the entire redesigned coding sequence of the target gene was obtained by fusing the above available two cDNA fragments with SOE-PCR (splicing by overlapping extension-PCR). We have cloned the human RANK gene (ten exons; CDS 1851 bp) using this strategy. The redesigned cDNA was transfected into an eukaryotic expression system (A459 cells) to verify its expression. RT-PCR and western blotting confirmed this. To validate our method, we also successfully cloned human TIMP2 gene (five exons; CDS 660 bp) also having a regionalized high GC content. Our strategy for combining codon optimization and SOE-PCR to clone difficult genes is thus feasible and potentially universally applicable.

  6. The Diabetes Evaluation Framework for Innovative National Evaluations (DEFINE): Construct and Content Validation Using a Modified Delphi Method.

    PubMed

    Paquette-Warren, Jann; Tyler, Marie; Fournie, Meghan; Harris, Stewart B

    2017-06-01

    In order to scale-up successful innovations, more evidence is needed to evaluate programs that attempt to address the rising prevalence of diabetes and the associated burdens on patients and the healthcare system. This study aimed to assess the construct and content validity of the Diabetes Evaluation Framework for Innovative National Evaluations (DEFINE), a tool developed to guide the evaluation, design and implementation with built-in knowledge translation principles. A modified Delphi method, including 3 individual rounds (questionnaire with 7-point agreement/importance Likert scales and/or open-ended questions) and 1 group round (open discussion) were conducted. Twelve experts in diabetes, research, knowledge translation, evaluation and policy from Canada (Ontario, Quebec and British Columbia) and Australia participated. Quantitative consensus criteria were an interquartile range of ≤1. Qualitative data were analyzed thematically and confirmed by participants. An importance scale was used to determine a priority multi-level indicator set. Items rated very or extremely important by 80% or more of the experts were reviewed in the final group round to build the final set. Participants reached consensus on the content and construct validity of DEFINE, including its title, overall goal, 5-step evaluation approach, medical and nonmedical determinants of health schematics, full list of indicators and associated measurement tools, priority multi-level indicator set and next steps in DEFINE's development. Validated by experts, DEFINE has the right theoretic components to evaluate comprehensively diabetes prevention and management programs and to support acquisition of evidence that could influence the knowledge translation of innovations to reduce the burden of diabetes. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  7. IMF-Therapy (Intention controlled Myo-Feedback)--an innovative method in the treatment of peripheral nerve lesions.

    PubMed

    Hall, K; Schmidt, U; Schmidhammer, R

    2007-01-01

    Physiotherapy is a well established part of the rehabilitation of peripheral nerve paralysis. The aim of this type of treatment is to re-establish arbitrary functions by improving the patients' active and passive mobility as well as their strength and stamina. IMF-Therapy (Intention controlled Myo-Feedback) is an innovative method in the treatment of peripheral nerve lesions that goes beyond the purely neuro-scientific framework and also takes into account methods and concepts of the psychology of learning. The essential assumption is that things learnt in the past are firmly established in the long term motor memory and can be reactivated by the patient. From results achieved in 32 patients treated with this therapy it can be concluded that IMF-Therapy may be a promising additional rehabilitation tool in peripheral nerve lesion.

  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. 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.

  10. An innovative method for preparation for rural practice: the high school rural scholars program.

    PubMed

    Crump, William J; Fricker, R Steve; Moore, Amanda N; Coakley, Virginia L

    2002-11-01

    Recent studies have shown that the most effective strategy to increase the number of practicing rural physicians is to admit more students with affinity for rural practice to medical school. A significant limitation of this strategy is the generally lower scores made on standardized tests by those from smaller high schools in rural areas. Although once admitted these students perform on a par with their urban counterparts, admission remains an obstacle for many. This report summarizes the second year of an innovative program that placed rising high school seniors in shadowing opportunities in their home towns while providing preparation for the ACT exam in a virtual classroom environment. Results show that the 20 students involved increased their knowledge of the strengths and weaknesses of health care in their towns as well as their understanding of how various health professionals function. The ACT composite score increased by 1.2 points, to 23. The reading and science subscores showed the largest increase. Although labor-intensive for the coordinators, the program was successful in meeting the stated goals.

  11. Strategies for Longitudinal Research with Youth in Foster Care: A Demonstration of Methods, Barriers, and Innovations.

    PubMed

    Jackson, Yo; Gabrielli, Joy; Tunno, Angela M; Hambrick, Erin P

    2012-07-01

    Longitudinal research on youth in foster care is important, but often challenging to accomplish. To assist the field, a thorough description of the development of the SPARK (Studying Pathways to Adjustment and Resilience in Kids) project, a longitudinal research project on the mechanisms of resilience for foster youth and their caregivers, is presented. Authors explain the difficult task for researchers in accessing youth in foster care and suggest strategies for success. Recruitment approaches for foster youth and their families are also provided along with examples of effective techniques. Data collection concerns are discussed, and the authors provide recommendations for researchers to consider when asking youth sensitive questions. Finally, data collection on academic information from teachers and how the SPARK project works with the academic community to gain information on school functioning for youth in the project is described. Suggestions for methodology utilized in future research along with examples of innovative adjustments to typical research procedures are provided as guidance for how research on maltreated youth can be conducted.

  12. Strategies for Longitudinal Research with Youth in Foster Care: A Demonstration of Methods, Barriers, and Innovations

    PubMed Central

    Jackson, Yo; Gabrielli, Joy; Tunno, Angela M.; Hambrick, Erin P.

    2012-01-01

    Longitudinal research on youth in foster care is important, but often challenging to accomplish. To assist the field, a thorough description of the development of the SPARK (Studying Pathways to Adjustment and Resilience in Kids) project, a longitudinal research project on the mechanisms of resilience for foster youth and their caregivers, is presented. Authors explain the difficult task for researchers in accessing youth in foster care and suggest strategies for success. Recruitment approaches for foster youth and their families are also provided along with examples of effective techniques. Data collection concerns are discussed, and the authors provide recommendations for researchers to consider when asking youth sensitive questions. Finally, data collection on academic information from teachers and how the SPARK project works with the academic community to gain information on school functioning for youth in the project is described. Suggestions for methodology utilized in future research along with examples of innovative adjustments to typical research procedures are provided as guidance for how research on maltreated youth can be conducted. PMID:22773879

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Tradition and Innovation: A New Way To Explore a TESOL Method.

    ERIC Educational Resources Information Center

    Liao, Xiaoqing

    This paper discusses a Teaching English to Speakers of Other Languages (TESOL) method appropriate for use in classrooms in China and offers a methodological framework with teaching principles. This method combines the communicative approach to language teaching with the more traditional grammar method. TESOL approaches in China are influenced by…

  18. Cloning in reproductive medicine.

    PubMed

    Illmensee, K

    2001-08-01

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

  19. National study of excellence and innovation in physical therapist education, Part 1-design, method, and results.

    PubMed

    Jensen, Gail M; Nordstrom, Terrence; Mostrom, Elizabeth; Hack, Laurita M; Gwyer, Janet

    2017-06-06

    The Carnegie Foundation for the Advancement of Teaching commissioned the Preparation for the Professions Program, a qualitative study of professional education in 5 professions: medicine, nursing, law, engineering and clergy. These studies identified curricular structures, instructional practices, assessment approaches and environmental characteristics that support the preparation of professionals and led to educational reforms. The physical therapy profession has not had any in-depth, national investigation of physical therapist education since the Catherine Worthingham studies conducted more than 50 years ago. This research was a Carnegie-type study, investigating elements of excellence and innovation in academic and clinical physical therapist education in the United States. Five physical therapist education researchers from across the United States used a qualitative multiple-case study design. Six academic and 5 clinical programs were selected for the study. The academic institutions and clinical agencies studied were diverse in size, institutional setting, geography, and role in residency education. Qualitative case studies were generated from review of artifacts, field observations, and interviews (individual and focus group), and they provided the data for the study. A conceptual framework grounded in 3 major dimensions was generated, with 8 supporting elements: (1) culture of excellence (shared beliefs and values, leadership and vision, drive for excellence, and partnerships), (2) praxis of learning (signature pedagogy, practice-based learning, creating adaptive learners, and professional formation); and (3) organizational structures and resources. Building on the work of the Carnegie's Preparation for the Professions Program, a conceptual model was developed, representing the dimensions and elements of excellence in physical therapist education that is centered on the foundational importance of a nexus of linked and highly valued aims of being learner

  20. Preparing Nurse Leaders to Innovate: Iowa's Innovation Seminar.

    PubMed

    Joseph, M Lindell; Rhodes, Ann; Watson, Carol A

    2016-02-01

    Currently, no doctoral guidelines to teach innovation exist. Using Christensen's theory of disruptive innovation, the five discovery skills used by disruptive innovators provide the framework for designing a leadership development approach to enable and support a mindset to innovate. Executive leadership students were provided with didactic content on innovation, were assigned to non-healthcare settings for an anthropological dig to uncover innovative activities, and were provided with reflective prompts to enable a new context for innovation. Faculty collaborated with other fields and took risks to provide new contexts to innovate. Students identified and proposed innovations for current health care issues. Some of the innovations included processes, evaluation methods, data analytics for care design, and patient engagement solutions. Faculty crossed borders for field experiences, as well as disciplinary borders. This collaborative seminar demonstrated that it is possible to develop executive nurse leaders to innovate. Copyright 2016, SLACK Incorporated.

  1. 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

  2. A Simple Method for the Extraction, PCR-amplification, Cloning, and Sequencing of Pasteuria 16S rDNA from Small Numbers of Endospores

    PubMed Central

    Atibalentja, N.; Noel, G. R.; Ciancio, A.

    2004-01-01

    For many years the taxonomy of the genus Pasteuria has been marred with confusion because the bacterium could not be cultured in vitro and, therefore, descriptions were based solely on morphological, developmental, and pathological characteristics. The current study sought to devise a simple method for PCR-amplification, cloning, and sequencing of Pasteuria 16S rDNA from small numbers of endospores, with no need for prior DNA purification. Results show that DNA extracts from plain glass bead-beating of crude suspensions containing 10,000 endospores at 0.2 × 10⁶ endospores ml-1 were sufficient for PCR-amplification of Pasteuria 16S rDNA, when used in conjunction with specific primers. These results imply that for P. penetrans and P. nishizawae only one parasitized female of Meloidogyne spp. and Heterodera glycines, respectively, should be sufficient, and as few as eight cadavers of Belonolaimus longicaudatus with an average number of 1,250 endospores of "Candidatus Pasteuria usgae" are needed for PCR-amplification of Pasteuria 16S rDNA. The method described in this paper should facilitate the sequencing of the 16S rDNA of the many Pasteuria isolates that have been reported on nematodes and, consequently, expedite the classification of those isolates through comparative sequence analysis. PMID:19262793

  3. An innovative method for extracting isotopic information from low-resolution gamma spectra

    SciTech Connect

    Miko, D.; Estep, R.J.; Rawool-Sullivan, M.W.

    1998-12-01

    A method is described for the extraction of isotopic information from attenuated gamma ray spectra using the gross-count material basis set (GC-MBS) model. This method solves for the isotopic composition of an unknown mixture of isotopes attenuated through an absorber of unknown material. For binary isotopic combinations the problem is nonlinear in only one variable and is easily solved using standard line optimization techniques. Results are presented for NaI spectrum analyses of various binary combinations of enriched uranium, depleted uranium, low burnup Pu, {sup 137}Cs, and {sup 133}Ba attenuated through a suite of absorbers ranging in Z from polyethylene through lead. The GC-MBS method results are compared to those computed using ordinary response function fitting and with a simple net peak area method. The GC-MBS method was found to be significantly more accurate than the other methods over the range of absorbers and isotopic blends studied.

  4. A restriction enzyme based cloning method to assess the in vitro replication capacity of HIV-1 subtype C Gag-MJ4 chimeric viruses.

    PubMed

    Claiborne, Daniel T; Prince, Jessica L; Hunter, Eric

    2014-08-31

    The protective effect of many HLA class I alleles on HIV-1 pathogenesis and disease progression is, in part, attributed to their ability to target conserved portions of the HIV-1 genome that escape with difficulty. Sequence changes attributed to cellular immune pressure arise across the genome during infection, and if found within conserved regions of the genome such as Gag, can affect the ability of the virus to replicate in vitro. Transmission of HLA-linked polymorphisms in Gag to HLA-mismatched recipients has been associated with reduced set point viral loads. We hypothesized this may be due to a reduced replication capacity of the virus. Here we present a novel method for assessing the in vitro replication of HIV-1 as influenced by the gag gene isolated from acute time points from subtype C infected Zambians. This method uses restriction enzyme based cloning to insert the gag gene into a common subtype C HIV-1 proviral backbone, MJ4. This makes it more appropriate to the study of subtype C sequences than previous recombination based methods that have assessed the in vitro replication of chronically derived gag-pro sequences. Nevertheless, the protocol could be readily modified for studies of viruses from other subtypes. Moreover, this protocol details a robust and reproducible method for assessing the replication capacity of the Gag-MJ4 chimeric viruses on a CEM-based T cell line. This method was utilized for the study of Gag-MJ4 chimeric viruses derived from 149 subtype C acutely infected Zambians, and has allowed for the identification of residues in Gag that affect replication. More importantly, the implementation of this technique has facilitated a deeper understanding of how viral replication defines parameters of early HIV-1 pathogenesis such as set point viral load and longitudinal CD4+ T cell decline.

  5. 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.

  6. A model for scale up of family health innovations in low-income and middle-income settings: a mixed methods study

    PubMed Central

    Bradley, Elizabeth H; Curry, Leslie A; Taylor, Lauren A; Pallas, Sarah Wood; Talbert-Slagle, Kristina; Yuan, Christina; Fox, Ashley; Minhas, Dilpreet; Ciccone, Dana Karen; Berg, David; Pérez-Escamilla, Rafael

    2012-01-01

    Background Many family health innovations that have been shown to be both efficacious and cost-effective fail to scale up for widespread use particularly in low-income and middle-income countries (LMIC). Although individual cases of successful scale-up, in which widespread take up occurs, have been described, we lack an integrated and practical model of scale-up that may be applicable to a wide range of public health innovations in LMIC. Objective To develop an integrated and practical model of scale-up that synthesises experiences of family health programmes in LMICs. Data sources We conducted a mixed methods study that included in-depth interviews with 33 key informants and a systematic review of peer-reviewed and grey literature from 11 electronic databases and 20 global health agency web sites. Study eligibility criteria, participants and interventions We included key informants and studies that reported on the scale up of several family health innovations including Depo-Provera as an example of a product innovation, exclusive breastfeeding as an example of a health behaviour innovation, community health workers (CHWs) as an example of an organisational innovation and social marketing as an example of a business model innovation. Key informants were drawn from non-governmental, government and international organisations using snowball sampling. An article was excluded if the article: did not meet the study's definition of the innovation; did not address dissemination, diffusion, scale up or sustainability of the innovation; did not address low-income or middle-income countries; was superficial in its discussion and/or did not provide empirical evidence about scale-up of the innovation; was not available online in full text; or was not available in English, French, Spanish or Portuguese, resulting in a final sample of 41 peer-reviewed articles and 30 grey literature sources. Study appraisal and synthesis methods We used the constant comparative method of

  7. 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…

  8. 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...

  9. 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...

  10. 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…

  11. 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...

  12. 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...

  13. 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

  14. A method for determining the unitary functional capacity of cloned channels and transporters expressed in Xenopus laevis oocytes.

    PubMed

    Zampighi, G A; Kreman, M; Boorer, K J; Loo, D D; Bezanilla, F; Chandy, G; Hall, J E; Wright, E M

    1995-11-01

    The Xenopus laevis oocyte is widely used to express exogenous channels and transporters and is well suited for functional measurements including currents, electrolyte and nonelectrolyte fluxes, water permeability and even enzymatic activity. It is difficult, however, to transform functional measurements recorded in whole oocytes into the capacity of a single channel or transporter because their number often cannot be estimated accurately. We describe here a method of estimating the number of exogenously expressed channels and transporters inserted in the plasma membrane of oocytes. The method is based on the facts that the P (protoplasmic) face in water-injected control oocytes exhibit an extremely low density of endogenous particles (212 +/- 48 particles/microns2, mean, SD) and that exogenously expressed channels and transporters increased the density of particles (up to 5,000/microns2) only on the P face. The utility and generality of the method were demonstrated by estimating the "gating charge" per particle of the Na+/glucose cotransporter (SGLT1) and a nonconducting mutant of the Shaker K+ channel proteins, and the single molecule water permeability of CHIP (Channel-like In-tramembrane Protein) and MIP (Major Intrinsic Protein). We estimated a "gating charge" of approximately 3.5 electronic charges for SGLT1 and approximately 9 for the mutant Shaker K+ channel from the ratio of Qmax to density of particles measured on the same oocytes. The "gating charges" were 3-fold larger than the "effective valences" calculated by fitting a Boltzmann equation to the same charge transfer data suggesting that the charge movement in the channel and cotransporter occur in several steps. Single molecule water permeabilities (pfs) of 1.4 x 10(-14) cm3/sec for CHIP and of 1.5 x 10(-16) cm3/sec for MIP were estimated from the ratio of the whole-oocyte water permeability (Pf) to the density of particles. Therefore, MIP is a water transporter in oocytes, albeit approximately 100

  15. Recombination-assisted megaprimer (RAM) cloning

    PubMed Central

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

    2014-01-01

    No molecular cloning technique is considered universally reliable, and many suffer from being too laborious, complex, or expensive. Restriction-free cloning is among the simplest, most rapid, and cost-effective methods, but does not always provide successful results. We modified this method to enhance its success rate through the use of exponential amplification coupled with homologous end-joining. This new method, recombination-assisted megaprimer (RAM) cloning, significantly extends the application of restriction-free cloning, and allows efficient vector construction with much less time and effort when restriction-free cloning fails to provide satisfactory results. The following modifications were made to the protocol:•Limited number of PCR cycles for both megaprimer synthesis and the cloning reaction to reduce error propagation.•Elimination of phosphorylation and ligation steps previously reported for cloning methods that used exponential amplification, through the inclusion of a reverse primer in the cloning reaction with a 20 base pair region of homology to the forward primer.•The inclusion of 1 M betaine to enhance both reaction specificity and yield. PMID:26150930

  16. 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

  17. [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.

  18. Quick and clean cloning.

    PubMed

    Thieme, Frank; Marillonnet, Sylvestre

    2014-01-01

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

  19. Generation of stable infectious clones of plant viruses by using Rhizobium radiobacter for both cloning and inoculation.

    PubMed

    Tuo, Decai; Fu, Lanlan; Shen, Wentao; Li, Xiaoying; Zhou, Peng; Yan, Pu

    2017-10-01

    A novel Rhizobium radiobacter (synonym Agrobacterium tumefaciens)-mediated approach was developed to generate stable infectious clones of plant viruses. This method uses R. radiobacter for both cloning and inoculation of infectious clones, bypassing the requirement of cloning in E. coli to avoid the instability. Only three steps are included in this method: (i) construct viral genome-encoding plasmids in vitro by one-step Gibson assembly; (ii) transform the assembled DNA products into R. radiobacter; (iii) inoculate plants with the R. radiobacter clones containing the viral genome. Stable infectious clones were obtained from two potyviruses papaya ringspot virus (PRSV) and papaya leaf distortion mosaic virus (PLDMV) using this method, whereas attempts utilizing "classical" E. coli cloning system failed repeatedly. This method is simple and efficient, and is promising for a wide application in generation of infectious clones of plant virus, especially for those which are instable in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. A model for scale up of family health innovations in low-income and middle-income settings: a mixed methods study.

    PubMed

    Bradley, Elizabeth H; Curry, Leslie A; Taylor, Lauren A; Pallas, Sarah Wood; Talbert-Slagle, Kristina; Yuan, Christina; Fox, Ashley; Minhas, Dilpreet; Ciccone, Dana Karen; Berg, David; Pérez-Escamilla, Rafael

    2012-01-01

    Many family health innovations that have been shown to be both efficacious and cost-effective fail to scale up for widespread use particularly in low-income and middle-income countries (LMIC). Although individual cases of successful scale-up, in which widespread take up occurs, have been described, we lack an integrated and practical model of scale-up that may be applicable to a wide range of public health innovations in LMIC. To develop an integrated and practical model of scale-up that synthesises experiences of family health programmes in LMICs. We conducted a mixed methods study that included in-depth interviews with 33 key informants and a systematic review of peer-reviewed and grey literature from 11 electronic databases and 20 global health agency web sites. We included key informants and studies that reported on the scale up of several family health innovations including Depo-Provera as an example of a product innovation, exclusive breastfeeding as an example of a health behaviour innovation, community health workers (CHWs) as an example of an organisational innovation and social marketing as an example of a business model innovation. Key informants were drawn from non-governmental, government and international organisations using snowball sampling. An article was excluded if the article: did not meet the study's definition of the innovation; did not address dissemination, diffusion, scale up or sustainability of the innovation; did not address low-income or middle-income countries; was superficial in its discussion and/or did not provide empirical evidence about scale-up of the innovation; was not available online in full text; or was not available in English, French, Spanish or Portuguese, resulting in a final sample of 41 peer-reviewed articles and 30 grey literature sources. We used the constant comparative method of qualitative data analysis to extract recurrent themes from the interviews, and we integrated these themes with findings from the

  1. An innovative method for coordinate measuring machine one-dimensional self-calibration with simplified experimental process.

    PubMed

    Fang, Cheng; Butler, David Lee

    2013-05-01

    In this paper, an innovative method for CMM (Coordinate Measuring Machine) self-calibration is proposed. In contrast to conventional CMM calibration that relies heavily on a high precision reference standard such as a laser interferometer, the proposed calibration method is based on a low-cost artefact which is fabricated with commercially available precision ball bearings. By optimizing the mathematical model and rearranging the data sampling positions, the experimental process and data analysis can be simplified. In mathematical expression, the samples can be minimized by eliminating the redundant equations among those configured by the experimental data array. The section lengths of the artefact are measured at arranged positions, with which an equation set can be configured to determine the measurement errors at the corresponding positions. With the proposed method, the equation set is short of one equation, which can be supplemented by either measuring the total length of the artefact with a higher-precision CMM or calibrating the single point error at the extreme position with a laser interferometer. In this paper, the latter is selected. With spline interpolation, the error compensation curve can be determined. To verify the proposed method, a simple calibration system was set up on a commercial CMM. Experimental results showed that with the error compensation curve uncertainty of the measurement can be reduced to 50%.

  2. 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.

  3. Human cloning and human dignity.

    PubMed

    Birnbacher, Dieter

    2005-03-01

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

  4. 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

  5. Influence of menstruation on the microbiota of healthy women's labia minora as analyzed using a 16S rRNA gene-based clone library method.

    PubMed

    Shiraishi, Tsukasa; Fukuda, Kazumasa; Morotomi, Nobuo; Imamura, Yuri; Mishima, Junko; Imai, Shigeo; Miyazawa, Kiyoshi; Taniguchi, Hatsumi

    2011-01-01

    The aim of this study was to determine the influence of menstruation on the bacterial population of healthy Japanese women's vulvas, especially the labia minora. Labia minora swabs were obtained from 10 premenopausal, nonpregnant Japanese women at premenstruation and on day 2 of menstruation. Vaginal swabs were also obtained from 3 out of the 10 women. No significant difference was found in the average bacterial cell count between the menstruation and premenstruation samples. Molecular analysis using a 16S rRNA gene-based clone library method detected 22 genera from the labia minora swabs (total 20), with the genus Lactobacillus being predominant at both premenstruation and during menstruation in 7 out of the 10 women. Of the other 3 women, 2 showed various kinds of bacterial species, including oral and fecal bacteria, with Atopobium vaginae and Gardnerella vaginalis predominating in the remaining woman's vulva in both conditions. In total, 6 out of 10 cases (60%) showed significantly different microbiota of the labia minora between the two conditions. These results imply that menstruation may promote a distortion of the bacterial flora around the vulva, although it causes no significant increase of the bacterial count.

  6. 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.

  7. An Innovative Method for Flood Peak Event Separation from Discharge Time Series

    NASA Astrophysics Data System (ADS)

    Borzì, Iolanda; Bonaccorso, Brunella; Viglione, Alberto; Tito Aronica, Giuseppe

    2017-04-01

    In extreme value theory, Peak Over Threshold (POT) method is recognized to make a better use of available data with respect to Block Maxima (BM) method, as it allows to retain all relevant observations for frequency analysis, provided that an appropriate threshold is selected. Definition of a proper threshold, able to identify independent events, is in general a critical issue. In addition, in many hydrological applications, such as flood frequency analysis, threshold levels are usually site-specific. This could be a strong limitation when comparing the flood responses of different river basins within a large geographical area. In the present study, an attempt is made to develop a relatively simple criterion for the determination of different thresholds to separate relevant flood events from a large dataset of daily discharge time series recorded at European scale. The proposed method arises from the theory of stochastic processes in the frequency domain, which can provide a meaningful description of the characteristics of the series. In particular, from the analysis of the power spectral density function and the signal of the process (e.g. narrowband or broadband), it is possible to define crossings (i.e. peaks) in clusters or independent crossings of the threshold. More specifically, such a selection process of significant events may help to understand whether relatively close peaks can be referred to a single flood event or independent events. Also in the frequency domain, a mean frequency of the random process can be defined for each series to be used as a possible common criterion for the definition of the different thresholds for flood peaks selection. The advantages of this method compared to other methods, such as traditional POT, lie in the fact that it establishes a robust procedure for the separation of independent events in time series, by means of the analysis in the frequency domain. Furthermore, the choice of the threshold for the selection of

  8. 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.

  9. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. 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 music…

  11. 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 music…

  12. 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…

  13. 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...

  14. 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…

  15. 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…

  16. An innovative SNP genotyping method adapting to multiple platforms and throughputs

    USDA-ARS?s Scientific Manuscript database

    Single nucleotide polymorphisms (SNPs) are highly abundant, distributed throughout the genome in various species, and therefore they are widely used as genetic markers. However, the usefulness of this genetic tool relies heavily on the availability of user-friendly SNP genotyping methods. We have d...

  17. 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...

  18. 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…

  19. Dilemmas in Educational Innovation: A Problem for Anthropology as Clinical Method.

    ERIC Educational Resources Information Center

    Smith, Louis M.

    Four school buildings encompassing a total of 11 open instructional areas were subjected to the clinical method or participant observation mode of inquiry. Two interrelated conclusions regarding the open space design resulted: (1) a distinguishable variation in administrative strategies existed when each group of teachers developed its own style,…

  20. Dry coating: an innovative enteric coating method using a cellulose derivative.

    PubMed

    Obara, S; Maruyama, N; Nishiyama, Y; Kokubo, H

    1999-01-01

    A novel enteric coating method was developed. This method involves direct feeding of coating polymer powder and simultaneous spraying of plasticizing agent, without either organic solvent or water, using a centrifugal granulator, fluidized bed, or tablet-coating machine. For film formation, a curing step was then necessary; this involved spraying a small amount (3-8% of core weight) of water or hydroxypropyl methylcellulose solution, followed by heating. Hydroxypropyl methylcellulose acetate succinate was used as the enteric coating polymer, and a combination of triethyl citrate and acetylated monoglyceride was used for plasticization. The coated beads and tablets were evaluated for gastric resistance, intestinal disintegration, and stability, in comparison with beads and tablets from a conventional aqueous coating with the same enteric polymer. The new method required a higher coating amount for gastric resistance compared with the conventional coating, but the processing time was dramatically reduced. The results show that this dry coating method is applicable to the preparation of enteric-coated beads and tablets using commercially available lab-scale apparatus.

  1. 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...

  2. 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...

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

    PubMed

    Kamath, Jagannath; Danda, Raja Shekar; Jayasheelan, Nikil; Singh, Rohit

    2016-06-01

    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. 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. 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. 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. This technique is dynamic, unique and accurate as compared to other methods for assessing mechanical axis deviation in a weight bearing position.

  4. Innovative method for the enrichment of high-polarity bioactive molecules present at low concentrations in complex matrices.

    PubMed

    Liu, Qing-Shan; He, Jie; Zhou, Wen-Bin; Gu, Yu-Long; Huang, Huoqiang; Li, Ke-Qin; Yin, Xiao-Ying

    2017-02-01

    Ginsenoside Rg1 is a valuable bioactive molecule but its high polarity and low concentration in complex mixtures makes it a challenge to separate Ginsenoside Rg1 from other saponins with similar structures, resulting in low extraction efficiency. The successful development of effective Rg1 molecularly imprinted polymers that exhibit high selectivity and adsorption may offer an improved method for the enrichment of active compounds. In this work, molecularly imprinted polymers were prepared with two different methods, precipitation polymerization or surface imprinted polymerization. Comparison of the adsorption abilities showed higher adsorption of the surface molecularly imprinted polymers prepared by surface imprinted polymerization, 46.80 mg/g, compared to the 27.74 mg/g observed for the molecularly imprinted polymers prepared by precipitation polymerization. Therefore, for higher adsorption of the highly polar Rg1, surface imprinted polymerization is a superior technique to make Rg1 molecularly imprinted polymers. The prepared surface molecularly imprinted polymers were tested as a solid-phase extraction column to directionally enrich Rg1 and its analogues from ginseng tea and total ginseng extracts. The column with surface molecularly imprinted polymers showed higher enrichment efficiency and better selectivity than a C18 solid-phase extraction column. Overall, a new, innovative method was developed to efficiently enrich high-polarity bioactive molecules present at low concentrations in complex matrices.

  5. An Innovative Non-Destructive and Computational Method for Uranium Activity and Enrichment Verification of UF{sub 6} Cylinder

    SciTech Connect

    El-Mongy, Sayed A.; Allam, K.M.; Farid, Osama M.

    2006-07-01

    Verification of {sup 235}U enrichment in uranium hexafluoride (UF{sub 6}) cylinders is often achieved by destructive and non-destructive assay techniques. These techniques are time consuming, need suitable and similar standard, in addition to loss of the nuclear material in the case of destructive analysis. This paper introduce an innovative approach for verifying of {sup 235}U enrichment in UF{sub 6} cylinder. The approach is based on measuring dose rate ({mu}Sv/h) resulted from the emitted gamma rays of {sup 235}U at the surface of the cylinder and then calculating the activity of uranium and enrichment percentage inside the cylinder by a three dimensional model. Attenuation of the main {sup 235}U gamma transitions due to the cylinder wall (5A Type of Ni alloy) was also calculated and corrected for. The method was applied on UF{sub 6} cylinders enriched with 19.75% of {sup 235}U. The calculated enrichment was found to be 18% with 9% uncertainty. By the suggested method, the calculated total uranium activity inside one of the investigated UF{sub 6} cylinder was found close to the target (certified) value (5.6 GBq) with 9% uncertainty. The method is being developed by taking into consideration other parameters. (authors)

  6. Investigation of innovative radiation imaging method and system for radiological environments

    NASA Astrophysics Data System (ADS)

    Nguyen, H.; Joung, J.; Kim, Y.; Lee, K.

    2017-03-01

    We have developed a novel imaging method that can be applied to most applications in the field of radiological environment imaging. It resolves either two-dimensional (2D) or three-dimensional (3D) distributions of radioactive sources in applications for homeland security, environmental monitoring, radiation contamination monitoring, baggage inspection, nuclear power plant monitoring, and more. The proposed imaging method uses a simple detector configured as a radiation-counting detector with spectroscopic capabilities. The detector module consists of two components: a flat field-of-view (FOV) collimator with a 30° FOV opening and a typical single-channel radiation detector made of a 2 in.×2 in. NaI(Tl) scintillator coupled to a 2 in photomultiplier tube (PMT). This simple detector module makes it possible to develop a cost-effective imaging system and provide design freedom in extending the system configuration to include one-dimensional (1D) or 2D detector-array shapes to meet the needs of various applications. One of most distinctive features of the new imaging method is that it uses only a pair of 2D projections to obtain a 3D reconstruction. The projections are measured by the proposed detector module at two positions orthogonal to one another; the measured projections are manipulated to enhance the resolution of the reconstructed 3D image. The imaging method comprises several steps performed consecutively: projection measurement, energy re-binning, projection separation, resolution and attenuation recovery, image reconstruction, and image consolidation and quantitative analysis. The resolution and attenuation recovery step provides the most distinctive and important processing by which the poor quality of projection data is enhanced. Such poor quality is mainly due to the use of a simple detector with a wide-opening flat FOV collimator. Simulation and experimental studies have been conducted to validate the proposed method. In this investigation, we

  7. Innovative Methods to Acquire and Adapt Soldier Skills (INMASS) in the Operational Environment

    DTIC Science & Technology

    2009-08-01

    multimedia exemplars included in the report are demonstrations of pilot approaches for Army consideration and support. Procedure: The research focused...their respect. 2) During a communication with members of the local government or HUMINT sources, Soldiers must empathize with community members to... pilot approaches for Army consideration and support. The exemplars provide a detailed look at each training method and explore how it might be used to

  8. Diffusion of innovation: enhancing the dissemination of the Ponseti method in Latin America through virtual forums.

    PubMed

    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.

  9. An Innovative Method for Monitoring Food Quality and the Healthfulness of Consumers’ Grocery Purchases

    PubMed Central

    Tran, Le-Thuy T.; Brewster, Philip J.; Chidambaram, Valliammai; Hurdle, John F.

    2017-01-01

    This study presents a method laying the groundwork for systematically monitoring food quality and the healthfulness of consumers’ point-of-sale grocery purchases. The method automates the process of identifying United States Department of Agriculture (USDA) Food Patterns Equivalent Database (FPED) components of grocery food items. The input to the process is the compact abbreviated descriptions of food items that are similar to those appearing on the point-of-sale sales receipts of most food retailers. The FPED components of grocery food items are identified using Natural Language Processing techniques combined with a collection of food concept maps and relationships that are manually built using the USDA Food and Nutrient Database for Dietary Studies, the USDA National Nutrient Database for Standard Reference, the What We Eat In America food categories, and the hierarchical organization of food items used by many grocery stores. We have established the construct validity of the method using data from the National Health and Nutrition Examination Survey, but further evaluation of validity and reliability will require a large-scale reference standard with known grocery food quality measures. Here we evaluate the method’s utility in identifying the FPED components of grocery food items available in a large sample of retail grocery sales data (~190 million transaction records). PMID:28475153

  10. Investigation of trend analysis of monthly total precipitation by an innovative method

    NASA Astrophysics Data System (ADS)

    Ay, Murat; Kisi, Ozgur

    2015-05-01

    The purpose of this study is to detect possible trend in monthly total precipitation rate on the basis of Mann-Kendall (MK) and a new method recently proposed by Şen trend tests. The new method is used for trend analysis of monthly total precipitation data recorded at Samsun, Trabzon, Ankara, Yozgat, Adana, and Antalya provinces in Turkey. For comparison purposes, the well-known MK trend test is also applied to the same data. According to the MK trend test, Samsun and Trabzon generally have statistically significant increasing trend. There is no statistically significant trend (trendless time series) for Ankara, Yozgat, and Adana provinces. While the February month of Antalya shows statistically significant decreasing trend, April month of Antalya shows statistically significant increasing trend. According to the Şen's trend test, peak and low values of monthly total precipitation of the six provinces show different trends (increasing, decreasing, or trendless time series) from the MK trend test. The study shows that the Şen method could be successfully used in evaluation of peak and low values of data for trend analysis of monthly total precipitation variable.

  11. Innovative method to suppress local geometry distortions for fabrication of interdigitated electrode arrays with nano gaps

    NASA Astrophysics Data System (ADS)

    Partel, S.; Urban, G.

    2016-03-01

    In this paper we present a method to optimize the lithography process for the fabrication of interdigitated electrode arrays (IDA) for a lift-off free electrochemical biosensor. The biosensor is based on amperometric method to allow a signal amplification by redox cycling. We already demonstrated a method to fabricate IDAs with nano gaps with conventional mask aligner lithography and two subsequent deposition processes. By decreasing the distance down to the nanometer range the linewidth variation is becoming the most critical factor and can result in a short circuit of the electrodes. Therefore, the light propagation and the resist pattern of the mask aligner lithography process are simulated to optimize the lithography process. To optimize the outer finger structure assistant features (AsFe) were introduced. The AsFe allow an optimization of the intensity distribution at the electrode fingers. Hence, the periodicity is expanded and the outer structure of the IDA is practically a part of the periodic array. The better CD uniformity can be obtained by adding three assistant features which generate an equal intensity distributions for the complete finger pattern. Considering a mask optimization of the outer structures would also be feasible. However, due to the strong impact of the gap between mask and wafer at contact lithography it is not practicable. The better choice is to create the same intensity distribution for all finger structures. With the introduction of the assistant features large areas with electrode gap sizes in the sub 100 nm region are demonstrated.

  12. An Innovative Method for Obtaining Consistent Images and Quantification of Histochemically Stained Specimens

    PubMed Central

    Sedgewick, Gerald J.; Ericson, Marna

    2015-01-01

    Obtaining digital images of color brightfield microscopy is an important aspect of biomedical research and the clinical practice of diagnostic pathology. Although the field of digital pathology has had tremendous advances in whole-slide imaging systems, little effort has been directed toward standardizing color brightfield digital imaging to maintain image-to-image consistency and tonal linearity. Using a single camera and microscope to obtain digital images of three stains, we show that microscope and camera systems inherently produce image-to-image variation. Moreover, we demonstrate that post-processing with a widely used raster graphics editor software program does not completely correct for session-to-session inconsistency. We introduce a reliable method for creating consistent images with a hardware/software solution (ChromaCal™; Datacolor Inc., NJ) along with its features for creating color standardization, preserving linear tonal levels, providing automated white balancing and setting automated brightness to consistent levels. The resulting image consistency using this method will also streamline mean density and morphometry measurements, as images are easily segmented and single thresholds can be used. We suggest that this is a superior method for color brightfield imaging, which can be used for quantification and can be readily incorporated into workflows. PMID:25575568

  13. 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"!

  14. 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

  15. 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

  16. Measuring tree height and preparation volume table using an innovative method.

    PubMed

    Lotfalian, Majid; Nouri, Zahra; Kooch, Yahya; Zobeiri, Mahmoud

    2007-10-15

    Zarbin (Cupressus sempervirence var. horizontalis) with its unique characteristics is one of the worthiest species which can be found in the central area of Alborz in the North of Iran. Especially in the Roodbar-manjil area, Chaloos-Hassanabad valley as well as it extends from Zarringol area to Gorgan. Although the distribution areas of this species have been protected, these forests have been invaded by the villagers who use this useful wood. For this reason in the Roodbar area, trees with DBH>30 cm are extremely rare. To recognize and to be aware of the stand quantity, the current research tries to calculate the species volume table in Roodbar area, to be the basis for any calculation of the volume of stand in the region. For this purpose, trees have been sampled using the line sampling method. After estimating the form factor, Tarif table have been prepared. In this study, a new method for measuring tree height is presented, in which, instead of measuring slope distance from observer to tree (which is difficult in young conifers because of existence branches in lower height) distance between the eye level of observer to tree butt is measured. Which doing of it is easier, time of field work is decreased and accuracy of measurement and calculation is increased.

  17. 'Twin2twin' an innovative method of empowering midwives to strengthen their professional midwifery organisations.

    PubMed

    Cadée, Franka; Perdok, Hilde; Sam, Betty; de Geus, Myrte; Kweekel, Liselotte

    2013-10-01

    midwives need professional support from a national midwifery organisation to be able to provide the services that are by regulatory mechanisms and accreditation expected of them. Not all midwives in the world are united in a professional organisation. The aim of this project was to strengthen the midwifery organisations of Sierra Leone and the Netherlands. During the process of the project it was realised that the development of a platform of exchange at organisational level would be enhanced by introducing personal exchange between individual midwives. In response to this new insight the original project plan was adjusted by incorporating the twin2twin method. twin2twin is a feminist methodology of mutual exchange between twenty pairs of midwives from different organisations (in this case Sierra Leone and the Netherlands). The method can be distinguished by 10 specific steps. It was developed, used and (re)evaluated through focus group discussions, storytelling and written evaluations. twinning of organisations was strengthened by adding a human component to the process. With the use of the 'twin2twin' method, midwives were encouraged to invested in a professional and personal bond with their 'twin sister'. This bond was independent and went beyond the relatively short four year project period. Through personal engagement and mutual exchange of knowledge and skills, midwives empowered each other to build and strengthen their midwifery organisations both in Sierra Leone and the Netherlands. (Empowerment refers to the expansion in people's ability to make strategic life choices in a context where this ability was previously denied to them (Narayan, 2005); organisational empowerment includes processes and structures that enhance members' skills and provides them with the mutual support necessary to effect community level change (Zimmerman, 1995).). despite challenges we are convinced that twin2twin can be of additional benefit for the success of other projects

  18. 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.

  19. An innovative method for automatic determination of time of arrival for Lamb waves excited by impact events

    NASA Astrophysics Data System (ADS)

    Zhu, Junxiao; Parvasi, Seyed Mohammad; Ho, Siu Chun Michael; Patil, Devendra; Ge, Maochen; Li, Hongnan; Song, Gangbing

    2017-05-01

    Lamb waves have great potential as a diagnostic tool in the application of structural health monitoring. Propagation properties of Lamb waves are affected by the state of the structure that the waves are traveling upon. Thus Lamb waves can carry information about the structure as they travel across a structure. However, the dispersive, multimodal and attenuation characteristics of Lamb waves make it difficult to determine the time of arrival of Lamb waves. To deal with these characteristics, an innovative method to automatically determine the time of arrival for impact-induced Lamb waves without human intervention is proposed in this paper. Lead zirconate titanate sensors mounted on the surface of an aluminum plate were used to measure the Lamb waves excited by an impact. The time of arrival was determined based on wavelet decomposition, Hilbert transform and statistics (Grubbs’ test and maximum likelihood estimation). Both of numerical analysis and physical measurements have verified the accuracy of this method for impacts on an aluminum plate.

  20. 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.

  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 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.

  3. Innovative Methods of Anthropogenic Landscape Reconstruction in the Urbanized Oil and Gas Region Environment

    NASA Astrophysics Data System (ADS)

    Shabatura, L. N.; Bauer, N. V.; Speranskaya, N. I.; Iatsevich, O. E.

    2016-10-01

    The article deals with the problems of the urbanized environment appearing as a result of intensive region developing. The state neglect towards people affects the population life quality of the of oil and gas extraction areas as well as problems resolving, and it provokes enormous losses for manufacturing and the whole region. The environment influences the person behaviour, one's perception and space understanding. The city environment is considered as the human existence space influencing on it directly, so it is necessary to renovate it. The authentic region developing cannot be reduced to "pure" economics (for not to be deserted), but needs to be fully mastered. To renovate the destroyed landscapes, it is necessary to use the landscape design methods making a cultural landscape. They help to increase the natural components of the city environment and to make it more harmonious, more harmless, more comfortable for residents.

  4. Innovative Method for Developing a Helium Pressurant Tank Suitable for the Upper Stage Flight Experiment

    NASA Technical Reports Server (NTRS)

    DeLay, Tom K.; Munafo, Paul (Technical Monitor)

    2001-01-01

    The AFRL USFE project is an experimental test bed for new propulsion technologies. It will utilize ambient temperature fuel and oxidizers (Kerosene and Hydrogen peroxide). The system is pressure fed, not pump fed, and will utilize a helium pressurant tank to drive the system. Mr. DeLay has developed a method for cost effectively producing a unique, large pressurant tank that is not commercially available. The pressure vessel is a layered composite structure with an electroformed metallic permeation barrier. The design/process is scalable and easily adaptable to different configurations with minimal cost in tooling development 1/3 scale tanks have already been fabricated and are scheduled for testing. The full-scale pressure vessel (50" diameter) design will be refined based on the performance of the sub-scale tank. The pressure vessels have been designed to operate at 6,000 psi. a PV/W of 1.92 million is anticipated.

  5. 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.

  6. Non-invasive evaluation of skin cytokines secretion: an innovative complementary method for monitoring skin disorders.

    PubMed

    Portugal-Cohen, Meital; Kohen, Ron

    2013-05-15

    In this review, a novel non-invasive approach based on skin surface wash sampling is described. Since the epidermis possesses a high metabolic activity, the secretion of various biomarkers can be exploited to develop non-invasive procedures for skin measurement to monitor disorders and to define a therapeutic strategy. Thus, we developed a method for the quantification of skin surface compounds. In this procedure, a well is placed on skin surface and is attached using an adhesive pad. Extraction buffer is introduced into the well for 30 min incubation period and the secretion of different biomarkers on skin surface can be measured: cytokines, antioxidants, peptides, RNA, DNA volatile organic compounds etc. Here, the focus is on cytokine measurement. After collecting skin samples cytokines can be quantified using ELISA assay. Since so far cytokine levels in skin have been evaluated mostly by invasive and prolonged procedures (punch biopsy, blister fluid and scrapping), employing this method has important implications, because it allows assessing cytokine amount with minimal invasion and high accuracy. We have already applied skin surface wash sampling for cytokine quantification in different clinical conditions: psoriasis, atopic dermatitis and chronic renal failure. A distinct pattern of cytokine secretion has been demonstrated for each disorder. Differences were also observed between lesional and non-lesional areas. The obtained results shed a new light on cutaneous cytokine expression in different clinical conditions. Moreover, the interplay between cytokines and other soluble compounds can give an added value in understanding the mechanism of skin pathologies. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Innovative method for optimizing Side-Scan Sonar mapping: The blind band unveiled

    NASA Astrophysics Data System (ADS)

    Pergent, Gérard; Monnier, Briac; Clabaut, Philippe; Gascon, Gilles; Pergent-Martini, Christine; Valette-Sansevin, Audrey

    2017-07-01

    Over the past few years, the mapping of Mediterranean marine habitats has become a priority for scientists, environment managers and stakeholders, in particular in order to comply with European directives (Water Framework Directive and Marine Strategy Framework Directive) and to implement legislation to ensure their conservation. Side-scan sonar (SSS) is recognised as one of the most effective tool for underwater mapping. However, interpretation of acoustic data (sonograms) requires extensive field calibration and the ground-truthing process remains essential. Several techniques are commonly used, with sampling methods involving grabs, scuba diving observations or Remotely Operated Vehicle (ROV) underwater video recordings. All these techniques are time consuming, expensive and only provide sporadic informations. In the present study, the possibility of coupling a camera with a SSS and acquiring underwater videos in a continuous way has been tested. During the 'PosidCorse' oceanographic survey carried out along the eastern coast of Corsica, optical and acoustic data were respectively obtained using a GoPro™ camera and a Klein 3000™ SSS. Thereby, five profiles were performed between 10 and 50 m depth, corresponding to more than 20 km of data acquisition. The vertical images recorded with the camera fixed under the SSS and positioned facing downwards provided photo mosaics of very good quality corresponding to the entire sonograms's blind band. From the photo mosaics, 94% of the different bottom types and main habitats have been identified; specific structures linked to hydrodynamics conditions, anthropic and biological activities have also been observed as well as the substrate on which the Posidonia oceanica meadow grows. The association between acoustic data and underwater videos has proved to be a non-destructive and cost-effective method for ground-truthing in marine habitats mapping. Nevertheless, in order to optimize the results over the next surveys

  8. 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.

  9. Nuclear transfer and cloning.

    PubMed

    Wolf, D P

    2001-10-01

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

  10. [Cloning--ethical aspects].

    PubMed

    Munzarová, M

    2004-01-01

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

  11. Innovative methods for parents and clinics to create tools for kids’ care (IMPACCT Kids’ Care) study protocol

    PubMed Central

    Angier, Heather; Marino, Miguel; Sumic, Aleksandra; O’Malley, Jean; Likumahuwa-Ackman, Sonja; Hoopes, Megan; Nelson, Christine; Gold, Rachel; Cohen, Deborah; Dickerson, Kristin; DeVoe, Jennifer E.

    2015-01-01

    Background Despite expansions in public health insurance, many children remain uninsured or experience gaps in coverage. Community health centers (CHCs) provide primary care to many children at risk for uninsurance and are well-positioned to help families obtain and retain children’s coverage. Recent advances in health information technology (HIT) capabilities provide the means to create tools that could enhance CHCs’ insurance outreach efforts. Objective To present the study design, baseline patient characteristics, variables, and statistical methods for the Innovative Methods for Parents And Clinics to Create Tools for Kids’ Care (IMPACCT Kids’ Care) study. Methods/design In this mixed methods study, we will design, test and refine health insurance outreach HIT tools through a user-centered process. We will then implement the tools in four CHCs and evaluate their effectiveness and barriers and facilitators to their implementation. To measure effectiveness, we will quantitatively assess health insurance coverage continuity and utilization of healthcare services for pediatric patients in intervention CHCs compared to matched control sites using electronic health record (EHR) and Oregon Medicaid administrative data over 18 months pre- and 18 months post-implementation (n = 34,867 children). We will also qualitatively assess the implementation process to understand how the tools fit into the clinics’ workflows and the CHC staff experiences with the tools. Conclusions This study creates, implements, and evaluates health insurance outreach HIT tools. The use of such tools will likely improve care delivery and health outcomes, reduce healthcare disparities for vulnerable populations, and enhance overall healthcare system performance. PMID:26291916

  12. 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

  13. Animal models of asthma: innovative methods of lung research and new pharmacological targets.

    PubMed

    Braun, Armin; Tschernig, Thomas

    2006-06-01

    Allergic diseases like bronchial asthma are increasing in societies with western lifestyle. In the last years substantial progress was made in the understanding of the underlying mechanisms and explanations like the hygiene hypothesis were developed. However the exact mechanisms of the physiological and immunological events in the lung leading to bronchial asthma are still not fully understood. Therefore, animal models of asthma have been established and improved to study the complex cellular interactions in vivo. Since mice became the most frequently used animal species the methods for detecting lung physiology, e.g. lung function measurements were adapted to the small size of the murine lung. Laser-dissection and precision cut lung slices have become common techniques to get a view into distinct lung compartments and cells. In addition genomic and proteomic approaches are now used widely. On the other hand a major conclusion of the workshop stated that more than one species is necessary in research and for pharmacological screening in asthma and COPD. The resulting new understanding in the mechanisms of asthma pathogenesis has lead to a rapid identification of novel pharmaceutical targets for treatment of the disease.

  14. Blogging as an Innovative Method of Peer-to-Peer Educational Sharing.

    PubMed

    Nedder, Melanie M; Levine, Sharon A; Galligan, Caroline; Avery, Kathleen Ryan; Eagan-Bengston, Elizabeth; Reilly, Karen M

    2017-02-01

    Nurses in the cardiac intensive care unit often attend professional education opportunities. However, it is difficult to share this information among nursing staff. Varying schedules, different shifts, and patient acuity limit the amount of time available for peer-to-peer sharing of educational information. A review of the literature revealed scant research on blogging for peer-to-peer education in general and particularly in nursing. To explore nurses' perception of the effectiveness of using a blog as a forum to provide peer-to-peer sharing of relevant professional education. Using a simple, free blogging website, the unit's nursing practice council developed a private blog for educational information sharing among the nursing staff. An online survey was administered to the unit's staff 15 months after the blog was implemented. Most respondents indicated that they thought the blog is an effective way to share professional education (86%), keeps them abreast of evidence-based practice (81%), and has led to practice change (59%). Nearly 80% of respondents agreed that they are more likely to attend professional conferences, and 62% would consider contributing blog posts. The survey results suggest that blogging may be an effective method of peer-to-peer sharing of education, although more rigorous research is required in this area. ©2017 American Association of Critical-Care Nurses.

  15. [Minimally invasive brush-biopsy: innovative method for early diagnosis of oral squamous cell carcinoma].

    PubMed

    Remmerbach, Torsten W; Hemprich, Alexander; Böcking, Alfred

    2007-01-01

    The aim of this prospective and blinded study was to investigate the diagnostic accuracy of conventional cytopathology of oral brush biopsies taken from suspicious oral lesions. In addition we checked slide based DNA image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA aneuploidy is a sensitive and specific marker for earliest detection of oral cancer using brush biopsies. Therefore the nuclear DNA contents were measured after Feulgen re-staining using a TV image analysis system. DNA aneuploidy was assumed if abnormal DNA stemlines or cells with DNA content greater 9c were observed. Sensitivity of our cytological diagnosis in oral smears for the detection of cancer cells thus was 91.3%, specificity for the detection of non-neoplastic cells was 95.1%, positive predictive value 94.4% and negative predictive value 92.3%. The adjuvant DNA image cytometry reached a sensitivity of 97.8%, the specificity and the positive predictive value were 100% and negative predictive value was 98.1%, respectively. Smears from oral brush biopsies of all visible oral lesions are an easily practicable, cheap, minimal invasive, painless and safe screening method for detection of oral precancerous lesions and squamous cell carcinomas in all stages. We conclude that DNA image cytometry is a very sensitive and highly specific, objective and reproducible adjuvant tool for identification of neoplastic cells in oral smears.

  16. An innovative strategy to clone positive modifier genes of defects caused by mtDNA mutations: MRPS18C as suppressor gene of m.3946G>A mutation in MT-ND1 gene.

    PubMed

    Rodríguez-García, María Elena; Cotrina-Vinagre, Francisco Javier; Carnicero-Rodríguez, Patricia; Martínez-Azorín, Francisco

    2017-07-01

    We have developed a new functional complementation approach to clone modifier genes which overexpression is able to suppress the biochemical defects caused by mtDNA mutations (suppressor genes). This strategy consists in transferring human genes into respiratory chain-deficient fibroblasts, followed by a metabolic selection in a highly selective medium. We used a normalized expression cDNA library in an episomal vector (pREP4) to transfect the fibroblasts, and a medium with glutamine and devoid of any carbohydrate source to select metabolically. Growing the patient's fibroblasts in this selective medium, the deficient cells rapidly disappear unless they are rescued by the cDNA of a suppressor gene. The use of an episomal vector allows us to carry out several rounds of transfection/selection (cyclical phenotypic rescue) to enrich the rescue with true clones of suppressor genes. Using fibroblasts from a patient with epileptic encephalopathy with the m.3946G>A (p.E214K) mutation in the MT-ND1 gene, several candidate genes were identified and one of them was characterized functionally. Thus, overexpression of MRPS18C gene (that encode for bS18m protein) suppressed the molecular defects produced by this mtDNA mutation, recovering the complex I activity and reducing the ROS produced by this complex to normal levels. We suggest that modulation of bS18m expression may be an effective therapeutic strategy for the patients with this mutation.

  17. Understanding clinical prediction models as 'innovations': a mixed methods study in UK family practice.

    PubMed

    Brown, Benjamin; Cheraghi-Sohi, Sudeh; Jaki, Thomas; Su, Ting-Li; Buchan, Iain; Sperrin, Matthew

    2016-08-09

    Well-designed clinical prediction models (CPMs) often out-perform clinicians at estimating probabilities of clinical outcomes, though their adoption by family physicians is variable. How family physicians interact with CPMs is poorly understood, therefore a better understanding and framing within a context-sensitive theoretical framework may improve CPM development and implementation. The aim of this study was to investigate why family physicians do or do not use CPMs, interpreting these findings within a theoretical framework to provide recommendations for the development and implementation of future CPMs. Mixed methods study in North West England that comprised an online survey and focus groups. One hundred thirty eight respondents completed the survey, which found the main perceived advantages to using CPMs were that they guided appropriate treatment (weighted rank [r] = 299; maximum r = 414 throughout), justified treatment decisions (r = 217), and incorporated a large body of evidence (r = 156). The most commonly reported barriers to using CPMs were lack of time (r = 163), irrelevance to some patients (r = 161), and poor integration with electronic health records (r = 147). Eighteen clinicians participated in two focus groups (i.e. nine in each), which revealed 13 interdependent themes affecting CPM use under three overarching domains: clinician factors, CPM factors and contextual factors. Themes were interdependent, indicating the tensions family physicians experience in providing evidence-based care for individual patients. The survey and focus groups showed that CPMs were valued when they supported clinical decision making and were robust. Barriers to their use related to their being time-consuming, difficult to use and not always adding value. Therefore, to be successful, CPMs should offer a relative advantage to current working, be easy to implement, be supported by training, policy and guidelines, and fit within the

  18. Transformation of anesthesia for ambulatory orthopedic surgery: A mixed-methods study of a diffusion of innovation in healthcare.

    PubMed

    Leggott, Kyle T; Martin, Matthew; Sklar, David; Helitzer, Deborah; Rosett, Randy; Crandall, Cameron; Vagh, Firoz; Mercer, Deana

    2016-09-01

    To provide insight into how an innovation in healthcare is implemented and diffused, we studied the transition from routine use of general anesthesia to peripheral nerve blocks (PNBs) for ambulatory orthopedic extremity surgery. Rogers' diffusion of innovations (DOI) theory was used as our theoretical framework. We identified themes that would be helpful for others attempting to diffuse innovations into healthcare settings. A mixed quantitative and qualitative methodology was used. We retrospectively reviewed operative and anesthesia records of patients who underwent ambulatory repair of distal radius fractures or arthroscopic knee meniscus procedures from 1998 to 2012 to identify whether general anesthetics or PNBs were used and the time course of the innovation. We interviewed orthopedic surgeons, anesthesiologists, and a nursing administrator working in the ambulatory surgery unit during the innovation to identify key themes associated with the adoption of PNBs. From 2003 to 2012, use of PNBs increased from less than 10% to 70% of cases studied. The adoption timeframe followed an S-shaped curve. Key themes included improved safety, quality, efficiency, physician leadership and trust, organizational structure, and technological change. The innovation involved an optional decision-making process and took root in a satellite facility and generally fit with Rogers DOI theory. The adoption and diffusion of PNBs provides a useful model for understanding innovations with optional decision-making in healthcare. Critical elements in our case were the characteristics of the innovation, which facilitated the decision-making process, and the positioning of the innovation in a peripheral structure away from core clinical facilities. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. An innovative method to produce green table olives based on "pied de cuve" technology.

    PubMed

    Martorana, Alessandra; Alfonzo, Antonio; Settanni, Luca; Corona, Onofrio; La Croce, Francesco; Caruso, Tiziano; Moschetti, Giancarlo; Francesca, Nicola

    2015-09-01

    The technology of "pied de cuve" (PdC) is applied in food process only to produce wines with an enriched community of pro-technological yeasts. PdC promotes the growth of the desirable microbial strains in a small volume of grape must acting as a starter inoculums for higher volumes. The aim of the present work was to investigate the use of partially fermented brines, a technology known as PdC, developed with lactic acid bacteria (LAB) on the microbiological, chemical and sensory characteristics of green fermented table olives during two consecutive campaigns. The experimental plan included two trials based on different PdCs: trial A, PdC obtained with Lactobacillus pentosus OM13; trial B, PdC obtained through a spontaneous fermentation. Two control additional trials without PdC were included for comparison: trial C, spontaneous fermentation; trial D, direct inoculation of L. pentosus OM13. The use of PdCs favoured the rapid increase of LAB concentrations in both trials A and B. These trials showed levels of LAB higher than trial C and almost superimposable to that of trial D. Trial B was characterized by a certain diversity of L. pentosus strains and some of them dominated the manufacturing process. These results indicated PdC as a valuable method to favour the growth of autochthonous L. pentosus strains. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) visibly discriminated olive processes fermented with the two experimental PdCs. Interestingly, on the basis of microbial and pH variables, both approaches showed that the olives produced with PdC technology are closely related to those of trial D, with the advantages of reducing the amount of starter to inoculate (trial A) and a higher LAB biodiversity (trial B). Volatile organic compound (VOC) composition and sensory analysis showed trials A and B different from the trials with no PdC added, in both years. Furthermore, the trial B showed the highest scores of green olive aroma and

  20. An innovative method to evaluate the suture compliance in sealing the surgical wound lips

    PubMed Central

    Saleh, Farid; Palmieri, Beniamino; Lodi, Danielle; Al-Sebeih, Khalid

    2008-01-01

    Background and aim: The increasing number of surgical procedures performed with local anesthesia, followed by immediate patient discharge from the hospital, emphasizes the need for a tight waterproof suture that is capable of maintaining its tensile strength in the postoperative phase when the wound tumescence, edema due to the anesthetic drug, and surgical trauma disappear. Moreover, the issue of having an accurate surgical wound closure is very relevant in vivo in order to prevent hemorrhage and exogenous microbial infections. This study aimed at designing a new a lab technique that could be used for evaluating the best surgical material. Using such a technique, we compared the wound-lip-sealing properties of three commonly-used suture threads, namely polyurethane, polypropylene, and polyamide. Materials and methods: The mechanical properties of same-size suture threads made from polyurethane, polypropylene, and polyamide, were compared in order to define the one that possess the best elastic properties by being able to counteract the tension-relaxation process in the first 12 hours following surgery. The tension holding capacity of the suture materials was measured in both in vivo and in vitro experiments. The surface area of the scar associated with the three different suture threads was measured and compared, and the permeability of the three different suture threads was assessed at 0 minute, 2 minute, 4 minute, 6 minute, and 8 minute- interval. Results: Results showed that polyurethane suture threads had significantly (P < 0.05) better tensile strength, elongation endurance before breakage, and better elasticity coefficient as compared to polypropylene and polyamide suture threads. Moreover, polyurethane suture threads were significantly (P < 0.05) more impermeable as compared to the other two suture thread types (polypropylene and polyamide). This impermeability was also associated with a tighter wound-lip-sealing ability, and with significantly (P < 0

  1. Multiple phase estimation in quantum cloning machines

    NASA Astrophysics Data System (ADS)

    Yao, Yao; Ge, Li; Xiao, Xing; Wang, Xiao-guang; Sun, Chang-pu

    2014-08-01

    Since the initial discovery of the Wootters-Zurek no-cloning theorem, a wide variety of quantum cloning machines have been proposed aiming at imperfect but optimal cloning of quantum states within its own context. Remarkably, most previous studies have employed the Bures fidelity or the Hilbert-Schmidt norm as the figure of merit to characterize the quality of the corresponding cloning scenarios. However, in many situations, what we truly care about is the relevant information about certain parameters encoded in quantum states. In this work, we investigate the multiple phase estimation problem in the framework of quantum cloning machines, from the perspective of quantum Fisher information matrix (QFIM). Focusing on the generalized d-dimensional equatorial states, we obtain the analytical formulas of QFIM for both universal quantum cloning machine (UQCM) and phase-covariant quantum cloning machine (PQCM), and prove that PQCM indeed performs better than UQCM in terms of QFIM. We highlight that our method can be generalized to arbitrary cloning schemes where the fidelity between the single-copy input and output states is input-state independent. Furthermore, the attainability of the quantum Cramér-Rao bound is also explicitly discussed.

  2. Innovative methods for parents and clinics to create tools for kids' care (IMPACCT Kids' Care) study protocol.

    PubMed

    Angier, Heather; Marino, Miguel; Sumic, Aleksandra; O'Malley, Jean; Likumahuwa-Ackman, Sonja; Hoopes, Megan; Nelson, Christine; Gold, Rachel; Cohen, Deborah; Dickerson, Kristin; DeVoe, Jennifer E

    2015-09-01

    Despite expansions in public health insurance, many children remain uninsured or experience gaps in coverage. Community health centers (CHCs) provide primary care to many children at risk for uninsurance and are well-positioned to help families obtain and retain children's coverage. Recent advances in health information technology (HIT) capabilities provide the means to create tools that could enhance CHCs' insurance outreach efforts. To present the study design, baseline patient characteristics, variables, and statistical methods for the Innovative Methods for Parents And Clinics to Create Tools for Kids' Care (IMPACCT Kids' Care) study. In this mixed methods study, we will design, test and refine health insurance outreach HIT tools through a user-centered process. We will then implement the tools in four CHCs and evaluate their effectiveness and barriers and facilitators to their implementation. To measure effectiveness, we will quantitatively assess health insurance coverage continuity and utilization of healthcare services for pediatric patients in intervention CHCs compared to matched control sites using electronic health record (EHR) and Oregon Medicaid administrative data over 18months pre- and 18months post-implementation (n=34,867 children). We will also qualitatively assess the implementation process to understand how the tools fit into the clinics' workflows and the CHC staff experiences with the tools. This study creates, implements, and evaluates health insurance outreach HIT tools. The use of such tools will likely improve care delivery and health outcomes, reduce healthcare disparities for vulnerable populations, and enhance overall healthcare system performance. ClinicalTrials.gov Identifier: NCT02298361. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. DESI-MS2: a rapid and innovative method for trace analysis of six cytostatic drugs in health care setting.

    PubMed

    Fabrizi, Giovanni; Fioretti, Marzia; Rocca, Lucia Mainero; Curini, Roberta

    2012-05-01

    With the aim of establishing exposure levels for hospital personnel preparing and administering cytostatic drugs (CDs), here, we present an innovative screening method based on the use of the desorption electrospray ionization (DESI) interface coupled with a hybrid quadrupole linear ion trap mass spectrometer. A rapid, simple, and sensitive procedure was developed for the simultaneous surface monitoring of cyclophosphamide, dacarbazine, methotrexate, vincristine, gemcitabine, and cytarabine. Since analytes were in the solid state, a novel approach based on the use of passive samplers was combined with the direct analysis of wipes. A PTFE-printed glass slide was used as a passive sampler, while hydrophobic centers of Swiffer® cloths were judged extremely efficient as wipe samplers. After the sampling period, the CD collectors were directly processed with the DESI-MS system without any further treatment. MS/MS confirmatory analysis was conducted using selected reaction monitoring in the positive ion mode and detection limits were evaluated. Values were at the picograms per square millimeter levels on the passive collector and at the picograms per square centimeter levels for the wipe ones. Direct determination on solid-state samples combined with mass spectrometry selectivity provided a powerful tool so far unapplied to occupational hygiene.

  4. Application of the ligation-independent cloning (LIC) method for rapid construction of a minigenome rescue system for Newcastle disease virus VG/GA strain

    USDA-ARS?s Scientific Manuscript database

    Newcastle disease virus (NDV) can cause serious diseases and substantial economic losses to the poultry industry. To gain a better understanding of NDV pathogenesis, several reverse genetics systems for different NDV strains have been established. However, the construction of infectious cDNA clone b...

  5. Technology Innovation

    EPA Pesticide Factsheets

    EPA produces innovative technologies and facilitates their creation in line with the Agency mission to create products such as the stormwater calculator, remote sensing, innovation clusters, and low-cost air sensors.

  6. Science Innovation

    EPA Pesticide Factsheets

    EPA provides innovative research activities that help transform the protection of human health and the environment with high-risk, high-reward Pathfinder Innovation Projects, the P3 student competition, and low-cost air monitoring.

  7. Research Innovation

    EPA Pesticide Factsheets

    EPA provides innovative research activities that help transform the protection of human health and the environment with high-risk, high-reward Pathfinder Innovation Projects, the P3 student competition, and low-cost air monitoring.

  8. Quick and Clean Cloning: A Ligation-Independent Cloning Strategy for Selective Cloning of Specific PCR Products from Non-Specific Mixes

    PubMed Central

    Thieme, Frank; Engler, Carola; Kandzia, Romy; Marillonnet, Sylvestre

    2011-01-01

    We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplification of the insert. The other side of the linearized cloning vector has homology with a sequence present in the insert, but nested and non-overlapping with the gene-specific primer used for amplification. Since only specific products contain this sequence, but none of the non-specific products, only specific products can be cloned. Cloning is performed using a one-step reaction that only requires incubation for 10 minutes at room temperature in the presence of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. The reaction mix is then directly transformed into E. coli where the annealed vector-insert complex is repaired and ligated. We have tested this method, which we call quick and clean cloning (QC cloning), for cloning of the variable regions of immunoglobulins expressed in non-Hodgkin lymphoma tumor samples. This method can also be applied to identify the flanking sequence of DNA elements such as T-DNA or transposon insertions, or be used for cloning of any PCR product with high specificity. PMID:21655102

  9. Quick and clean cloning: a ligation-independent cloning strategy for selective cloning of specific PCR products from non-specific mixes.

    PubMed

    Thieme, Frank; Engler, Carola; Kandzia, Romy; Marillonnet, Sylvestre

    2011-01-01

    We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplification of the insert. The other side of the linearized cloning vector has homology with a sequence present in the insert, but nested and non-overlapping with the gene-specific primer used for amplification. Since only specific products contain this sequence, but none of the non-specific products, only specific products can be cloned. Cloning is performed using a one-step reaction that only requires incubation for 10 minutes at room temperature in the presence of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. The reaction mix is then directly transformed into E. coli where the annealed vector-insert complex is repaired and ligated. We have tested this method, which we call quick and clean cloning (QC cloning), for cloning of the variable regions of immunoglobulins expressed in non-Hodgkin lymphoma tumor samples. This method can also be applied to identify the flanking sequence of DNA elements such as T-DNA or transposon insertions, or be used for cloning of any PCR product with high specificity.

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

    PubMed

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

    2010-01-08

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation. Copyright 2010 Elsevier Inc. All rights reserved.

  11. Cloning Mice and Men: Prohibiting the Use of iPS Cells for Human Reproductive Cloning

    PubMed Central

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

    2014-01-01

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation. PMID:20085739

  12. Cloning, killing, and identity.

    PubMed Central

    McMahan, J

    1999-01-01

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

  13. 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…

  14. 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…

  15. 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

  16. Innovation network.

    PubMed

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

    2016-10-11

    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.

  17. 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.

  18. 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 Central

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

    2013-01-01

    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. PMID:23618925

  19. 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

  20. Tracking the Evolution of Code Clones

    NASA Astrophysics Data System (ADS)

    Bakota, Tibor

    It is believed by many academic and industrial experts, that source code cloning (copy&paste programming) represents a significant threat to maintainability in an evolving software system. The real threat does not lie in the existence of duplications, but the fears are in connection with their evolution. There exist an abundance of algorithms for finding code clones in one particular version of a software system, but eliminating or even evaluating these clones often seems hopeless, as there may exist several thousands of them. Tracking the evolution of individual clones might solve the problem, as it could help identifying the inconsistently changing duplications: the clones which are really dangerous at a particular moment. In this paper we present an approach for mapping code duplications from one particular version of the software to another one, based on a similarity distance function. For the suspicious evolution patterns we introduce the term of "clone smells". By defining the relevant categories of the possible evolution patterns, the proposed method also gives a clue about why the reported code fragments might be dangerous. In the case study, clone smells were extracted, evaluated, and manually categorized throughout many versions of the jEdit system. The findings suggest that roughly half of the reported smells refer to inconsistent changes in the code.

  1. 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.

  2. 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)

  3. 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)

  4. Statement on Human Cloning

    MedlinePlus

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

  5. Statement on Human Cloning

    MedlinePlus

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

  6. 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.

  7. Advancing Clouds Lifecycle Representation in Numerical Models Using Innovative Analysis Methods that Bridge ARM Observations and Models Over a Breadth of Scales

    SciTech Connect

    Kollias, Pavlos

    2016-09-06

    This the final report for the DE-SC0007096 - Advancing Clouds Lifecycle Representation in Numerical Models Using Innovative Analysis Methods that Bridge ARM Observations and Models Over a Breadth of Scales - PI: Pavlos Kollias. The final report outline the main findings of the research conducted using the aforementioned award in the area of cloud research from the cloud scale (10-100 m) to the mesoscale (20-50 km).

  8. [A method using long primers for cloning the upstream sequence of delta-6 fatty acid desaturases gene of Thamnidium elegans by nested inverse PCR].

    PubMed

    Wang, De-Pei; Sun, Wei; Li, Ming-Chun; Wei, Dong-Sheng; Zhang, Ying-Hui; Xing, Lai-Jun

    2006-07-01

    Thamnidium elegans is a kind of phycomycete that produces essential unsaturated fatty acids, particularly y-linolenic acid. In this process, delta6-Fatty acid desaturase (D6D) plays a key role due to its enzymatic properties that catalyze the delta6 site dehydrogenation of precursor linoleic acid (18:2delta(9, 12) n-6) and a-linolenic acid (18:3delta(9, 12, 15) n-3). This reaction is the first and rate-limiting step of highly unsaturated fatty acids (HUFA) synthesis pathways. After we have isolated and cloned the gene coding delta6-fatty acid desaturase from Thamnidium elegans As3.2806 (GenBank accession number DQ099380), our interest focuses on the promotion and regulation of the gene transcription. To achieve this aim, we designed long primers and used nested inverse PCR to amplify DNA flanking sequences. First, genome of Thamnidium elegans was extracted and digested with restriction enzymes EcoR I and Kpn I , respectively. Then we ligated the digested DNA with T4 ligase at low concentration which is propitious for linear DNA to joint intromolecule. According to the sequence of delta6-fatty acid desaturase gene of Thamnidium elegans, we designed a couple of 35nt long inverse primers and two couples of shorter inverse primers for inverse PCR. Three rounds of PCR reactions were performed. In the primary reaction, the ligated DNA was used as a template, and the product was used as the template of the secondary reaction, the tertiary reaction was achieved in the same way. After all the three rounds of reactions, we got a nice product about 4 kb from the EcoR I digested sample, in which a 1.3kb 5' upstream sequence (GenBank accession number DQ309425) of delta6-fatty acid desaturase gene containing several putative regulatory elements including TATA. box, FSE-2, AP-1 sites, CCAAT cis-element site and STRE-binding site was derived after sequencing. All of these implied intensely that this 1.3kb fragment is a condition-regulated promoter. It is the first report about

  9. 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…

  10. An Innovative Method for Evaluating Strategic Goals in a Public Agency: Conservation Leadership in the U.S. Forest Service

    Treesearch

    David N. Bengston; David P. Fan

    1999-01-01

    This article presents an innovative methodology for evaluating strategic planning goals in a public agency. Computer-coded content analysis was used to evaluate attitudes expressed in about 28,000 on-line news media stories about the U.S. Department of Agriculture Forest Service and its strategic goal of conservation leadership. Three dimensions of conservation...

  11. 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…

  12. Comparison of irradiated 15Kh2MFA material mechanical properties using conventional testing methods and innovative approach of small punch testing (SPT) and automated ball indentation (ABIT)

    NASA Astrophysics Data System (ADS)

    Kopriva, R.; Petelova, P.; Eliasova, I.; Kytka, M.; Culek, M.

    2017-02-01

    Article describes two innovative testing methods - Small Punch Testing (SPT) and Automated Ball Indentation Test (ABIT) - which are based on the determination and evaluation of material properties from miniaturized testing specimens. These methods are very promising due to minimum material needed for testing and also in case of testing highly irradiated materials of components that are not included in standard surveillance programs. The test results were obtained for reactor pressure vessel (RPV) base material 15Ch2MFA in both states - initial unirradiated and irradiated. Subsequently results were compared with standard tensile tests to prove applicability of these testing methods for the evaluation of degradation of irradiated structural materials of nuclear power plants.

  13. Creating a testing field where delta technology and water innovations are tested and demonstrated with the help of citizen science methods

    NASA Astrophysics Data System (ADS)

    de Vries, Sandra; Rutten, Martine; de Vries, Liselotte; Anema, Kim; Klop, Tanja; Kaspersma, Judith

    2017-04-01

    In highly populated deltas, much work is to be done. Complex problems ask for new and knowledge driven solutions. Innovations in delta technology and water can bring relief to managing the water rich urban areas. Testing fields form a fundamental part of the knowledge valorisation for such innovations. In such testing fields, product development by start-ups is coupled with researchers, thus supplying new scientific insights. With the help of tests, demonstrations and large-scale applications by the end-users, these innovations find their way to the daily practices of delta management. More and more cities embrace the concept of Smart Cities to tackle the ongoing complexity of urban problems and to manage the city's assets - such as its water supply networks and other water management infrastructure. Through the use of new technologies and innovative systems, data are collected from and with citizens and devices - then processed and analysed. The information and knowledge gathered are keys to enabling a better quality of life. By testing water innovations together with citizens in order to find solutions for water management problems, not only highly spatial amounts of data are provided by and/or about these innovations, they are also improved and demonstrated to the public. A consortium consisting of a water authority, a science centre, a valorisation program and two universities have joined forces to create a testing field for delta technology and water innovations using citizen science methods. In this testing field, the use of citizen science for water technologies is researched and validated by facilitating pilot projects. In these projects, researchers, start-ups and citizens work together to find the answer to present-day water management problems. The above mentioned testing field tests the use of crowd-sourcing data as for example hydrological model inputs, or to validate remote sensing applications, or improve water management decisions. Currently the

  14. OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments

    PubMed Central

    Liu, Chang-Jun; Jiang, Hui; Wu, Lei; Zhu, Ling-Yun; Meng, Er; Zhang, Dong-Yi

    2017-01-01

    Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on Overlap Extension PCR and Recombination in vivo). OEPR cloning is a seamless, restriction- and ligation-independent method. The method takes advantage of both homologous recombination enzymes in E. coli and overlap PCR. Using OEPR cloning, a long fragment (1–6 kb) or multiple fragments (2–4 fragments) can be easily constructed and simultaneously assembled into a target vector. PMID:28300166

  15. 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.

  16. Isolation of exons from cloned DNA by exon trapping.

    PubMed

    Nisson, P E; Watkins, P C; Krizman, D B

    2001-05-01

    Exon trapping is an RNA polymerase chain reaction (PCR) method to clone expressed sequences or exons directly from mammalian genomic DNA. The basic protocol in this unit describes the method for trapping internal exons from cosmid clones and the second basic protocol describes trapping of 3 terminal exons. An describes 3 terminal exon trapping, which avoids subcloning of target DNA by ligating it to the vector for direct transfection. A describes a rapid cloning procedure using uracil DNA glycosylase.

  17. 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.

  18. 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.

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

    PubMed

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

    1989-09-01

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

  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. 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. Extremal quantum cloning machines

    NASA Astrophysics Data System (ADS)

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

    2005-10-01

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

  3. 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.

  4. Emotional reactions to human reproductive cloning.

    PubMed

    May, Joshua

    2016-01-01

    Extant surveys of people's attitudes towards human reproductive cloning focus on moral judgements alone, not emotional reactions or sentiments. This is especially important given that some (especially Leon Kass) have argued against such cloning on the ground that it engenders widespread negative emotions, like disgust, that provide a moral guide. To provide some data on emotional reactions to human cloning, with a focus on repugnance, given its prominence in the literature. This brief mixed-method study measures the self-reported attitudes and emotions (positive or negative) towards cloning from a sample of participants in the USA. Most participants condemned cloning as immoral and said it should be illegal. The most commonly reported positive sentiment was by far interest/curiosity. Negative emotions were much more varied, but anxiety was the most common. Only about a third of participants selected disgust or repugnance as something they felt, and an even smaller portion had this emotion come to mind prior to seeing a list of options. Participants felt primarily interested and anxious about human reproductive cloning. They did not primarily feel disgust or repugnance. This provides initial empirical evidence that such a reaction is not appropriately widespread. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  5. 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

  6. Surpassing the no-cloning limit with a heralded hybrid linear amplifier for coherent states

    NASA Astrophysics Data System (ADS)

    Haw, Jing Yan; Zhao, Jie; Dias, Josephine; Assad, Syed M.; Bradshaw, Mark; Blandino, Rémi; Symul, Thomas; Ralph, Timothy C.; Lam, Ping Koy

    2016-10-01

    The no-cloning theorem states that an unknown quantum state cannot be cloned exactly and deterministically due to the linearity of quantum mechanics. Associated with this theorem is the quantitative no-cloning limit that sets an upper bound to the quality of the generated clones. However, this limit can be circumvented by abandoning determinism and using probabilistic methods. Here, we report an experimental demonstration of probabilistic cloning of arbitrary coherent states that clearly surpasses the no-cloning limit. Our scheme is based on a hybrid linear amplifier that combines an ideal deterministic linear amplifier with a heralded measurement-based noiseless amplifier. We demonstrate the production of up to five clones with the fidelity of each clone clearly exceeding the corresponding no-cloning limit. Moreover, since successful cloning events are heralded, our scheme has the potential to be adopted in quantum repeater, teleportation and computing applications.

  7. Construction of an agroinfectious clone of bean rugose mosaic virus using Gibson Assembly.

    PubMed

    Bijora, Taise; Blawid, Rosana; Costa, Danielle K T; Aragão, Francisco J L; Souto, Eliezer R; Nagata, Tatsuya

    2017-03-18

    Construction of agroinfectious viral clones usually requires many steps of cloning and sub-cloning and also a binary vector, which makes the process laborious, time-consuming, and frequently susceptible to some degree of plasmid instability. Nowadays, novel methods have been applied to the assembly of infectious viral clones, and here we have applied isothermal, single-step Gibson Assembly (GA) to construct an agroinfectious clone of Bean rugose mosaic virus (BRMV) using a small binary vector. The procedure has drastically reduced the cloning steps, and BRMV could be recovered from agroinfiltrated common bean twenty days after inoculation, indicating that the infectious clone could spread in the plant tissues and efficiently generate a systemic infection. The virus was also recovered from leaves of common bean and soybean cultivars mechanically inoculated with infectious clone two weeks after inoculation, confirming the efficiency of GA cloning procedure to produce the first BRMV agroinfectious clone to bean and soybean.

  8. Surpassing the no-cloning limit with a heralded hybrid linear amplifier for coherent states

    PubMed Central

    Haw, Jing Yan; Zhao, Jie; Dias, Josephine; Assad, Syed M.; Bradshaw, Mark; Blandino, Rémi; Symul, Thomas; Ralph, Timothy C.; Lam, Ping Koy

    2016-01-01

    The no-cloning theorem states that an unknown quantum state cannot be cloned exactly and deterministically due to the linearity of quantum mechanics. Associated with this theorem is the quantitative no-cloning limit that sets an upper bound to the quality of the generated clones. However, this limit can be circumvented by abandoning determinism and using probabilistic methods. Here, we report an experimental demonstration of probabilistic cloning of arbitrary coherent states that clearly surpasses the no-cloning limit. Our scheme is based on a hybrid linear amplifier that combines an ideal deterministic linear amplifier with a heralded measurement-based noiseless amplifier. We demonstrate the production of up to five clones with the fidelity of each clone clearly exceeding the corresponding no-cloning limit. Moreover, since successful cloning events are heralded, our scheme has the potential to be adopted in quantum repeater, teleportation and computing applications. PMID:27782135

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

    PubMed

    Isasi, Rosario M; Annas, George J

    2006-01-01

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

  10. Experimental validation of an innovative deflection compensation method in a multi-beamlet negative-ion accelerator

    NASA Astrophysics Data System (ADS)

    Chitarin, G.; Kojima, A.; Agostinetti, P.; Aprile, D.; Baltador, C.; Hiratsuka, J.; Ichikawa, M.; Marconato, N.; Sartori, E.; Serianni, G.; Veltri, P.; Yoshida, M.; Kashiwagi, M.; Hanada, M.; Antoni, V.

    2017-08-01

    An innovative magnetic configuration called ADCM (Asymmetric Deflection Compensation Magnets) has been recently proposed for the cancellation of the "criss-cross" deflection of ion trajectories in multi-beamlet negative ion accelerators. Such undesired deflection occurs due to the transverse magnetic field, which is necessary for the suppression of co-extracted electrons. In view of the application of this concept to the ITER Heating Neutral Beam Injector, the new configuration has been experimentally tested for the first time in 2016, in the framework of a research collaboration between Consorzio RFX and QST.

  11. 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.

  12. 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

    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.

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

    PubMed Central

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

    2017-01-01

    Attempts at cloning a quantum system result in the introduction of imperfections in the state of the copies. This is a consequence of the no-cloning theorem, which is a fundamental law of quantum physics and the backbone of security for quantum communications. Although perfect copies are prohibited, a quantum state may be copied with maximal accuracy via various optimal cloning schemes. Optimal quantum cloning, which lies at the border of the physical limit imposed by the no-signaling theorem and the Heisenberg uncertainty principle, has been experimentally realized for low-dimensional photonic states. However, an increase in the dimensionality of quantum systems is greatly beneficial to quantum computation and communication protocols. Nonetheless, no experimental demonstration of optimal cloning machines has hitherto been shown for high-dimensional quantum systems. We perform optimal cloning of high-dimensional photonic states by means of the symmetrization method. We show the universality of our technique by conducting cloning of numerous arbitrary input states and fully characterize our cloning machine by performing quantum state tomography on cloned photons. In addition, a cloning attack on a Bennett and Brassard (BB84) quantum key distribution protocol is experimentally demonstrated to reveal the robustness of high-dimensional states in quantum cryptography. PMID:28168219

  14. 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?

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

    PubMed

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

    2017-02-01

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

  16. 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)

  17. 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.

  18. 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)

  19. Unit cloning and amplification as novel and universal strategies for complex vector construction and small DNA fragment preparation.

    PubMed

    Ye, Chunjiang; Gu, Jingsong; Chen, Sunxiao; Deng, Anmei; Li, Yue Zhong; Li, Dianxiang

    2010-09-01

    With a novel and universal strategy for the cloning of multiple DNA fragments, a complex synthetic vector (pVEC100), harboring the target DNA fragments in conventional 100-bp DNA ladder, was constructed for efficient and large-scale production of 100-bp DNA marker through bacteria fermentation, plasmid extraction and restrictive digestion. Since the restrictive digestion of complex vectors yields insufficient small DNA fragments, an innovative PCR model was developed as an alternative. The PCR model comprised a specially designed template vector and a unit amplification model for producing groups of small DNA fragments. The unit amplification model improved the efficiency of the PCR protocol and made it more economical and easier for small DNA fragment amplification. The approach presented in this paper--a unit cloning model for constructing complex synthetic vectors combined with the modular design of unit amplification by PCR--is a powerful method for preparing small DNA fragments of DNA molecular weight standards.

  20. 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

  1. Innovation Awards

    EPA Pesticide Factsheets

    EPA promotes environmental stewardship by recognizing innovators in schools, communities and businesses in categories such as environmental education, green chemistry, smart growth, green power, and reducing air pollution and climate change impacts.

  2. 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)

  3. 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)

  4. [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.

  5. An innovative method to reduce count loss from pulse pile-up in a photon-counting pixel for high flux X-ray applications

    NASA Astrophysics Data System (ADS)

    Lee, D.; Lim, K.; Park, K.; Lee, C.; Alexander, S.; Cho, G.

    2017-03-01

    In this study, an innovative fast X-ray photon-counting pixel for high X-ray flux applications is proposed. A computed tomography system typically uses X-ray fluxes up to 108 photons/mm2/sec at the detector and thus a fast read-out is required in order to process individual X-ray photons. Otherwise, pulse pile-up can occur at the output of the signal processing unit. These superimposed signals can distort the number of incident X-ray photons leading to count loss. To minimize such losses, a cross detection method was implemented in the photon-counting pixel. A maximum count rate under X-ray tube voltage of 90 kV was acquired which reflect electrical test results of the proposed photon counting pixel. A maximum count of 780 kcps was achieved with a conventional photon-counting pixel at the pulse processing time of 500 ns, which is the time for a pulse to return to the baseline from the initial rise. In contrast, the maximum count of about 8.1 Mcps was achieved with the proposed photon-counting pixel. From these results, it was clear that the maximum count rate was increased by approximately a factor 10 times by adopting the cross detection method. Therefore, it is an innovative method to reduce count loss from pulse pile-up in a photon-counting pixel while maintaining the pulse processing time.

  6. The influence of alternative plant propagation and stand establishment techniques on survival and growth of eastern cottonwood (Populus deltoids Bartr.) clones

    Treesearch

    Donald J. Kaczmarek; Randall Rousseau; Jeff A. Wright; Brian Wachelka

    2014-01-01

    Four eastern cottonwood clones, including standard operational clone ST66 and three advanced clonal selections were produced and included in a test utilizing five different plant propagation methods. Despite relatively large first-year growth differences among clones, all clones demonstrated similar responses to the treatments and clone × cutting treatment interactions...

  7. Cloning Components of Human Telomerase.

    DTIC Science & Technology

    1998-07-01

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

  8. Water relations of populus clones

    SciTech Connect

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

    1981-02-01

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

  9. 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.'

  10. [Cloning and sequence analysis of the DHBV genome of the brown ducks in Guilin region and establishment of the quantitative method for detecting DHBV].

    PubMed

    Su, He-Ling; Huang, Ri-Dong; He, Song-Qing; Xu, Qing; Zhu, Hua; Mo, Zhi-Jing; Liu, Qing-Bo; Liu, Yong-Ming

    2013-03-01

    Brown ducks carrying DHBV were widely used as hepatitis B animal model in the research of the activity and toxicity of anti-HBV dugs. Studies showed that the ratio of DHBV carriers in the brown ducks in Guilin region was relatively high. Nevertheless, the characters of the DHBV genome of Guilin brown duck remain unknown. Here we report the cloning of the genome of Guilin brown duck DHBV and the sequence analysis of the genome. The full length of the DHBV genome of Guilin brown duck was 3 027bp. Analysis using ORF finder found that there was an ORF for an unknown peptide other than S-ORF, PORF and C-ORF in the genome of the DHBV. Vector NTI 8. 0 analysis revealed that the unknown peptide contained a motif which binded to HLA * 0201. Aligning with the DHBV sequences from different countries and regions indicated that there were no obvious differences of regional distribution among the sequences. A fluorescence quantitative PCR for detecting DHBV was establishment based on the recombinant plasmid pGEM-DHBV-S constructed. This study laid the groundwork for using Guilin brown duck as a hepatitis B animal model.

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

    PubMed

    Mieszczanek, J; Kofta, W; Wedrychowicz, H

    2000-12-01

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

  12. Three concepts of cloning in human beings.

    PubMed

    Cui, Ke-Hui

    2005-07-01

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

  13. Quantifying innovation in surgery.

    PubMed

    Hughes-Hallett, Archie; Mayer, Erik K; Marcus, Hani J; Cundy, Thomas P; Pratt, Philip J; Parston, Greg; Vale, Justin A; Darzi, Ara W

    2014-08-01

    The objectives of this study were to assess the applicability of patents and publications as metrics of surgical technology and innovation; evaluate the historical relationship between patents and publications; develop a methodology that can be used to determine the rate of innovation growth in any given health care technology. The study of health care innovation represents an emerging academic field, yet it is limited by a lack of valid scientific methods for quantitative analysis. This article explores and cross-validates 2 innovation metrics using surgical technology as an exemplar. Electronic patenting databases and the MEDLINE database were searched between 1980 and 2010 for "surgeon" OR "surgical" OR "surgery." Resulting patent codes were grouped into technology clusters. Growth curves were plotted for these technology clusters to establish the rate and characteristics of growth. The initial search retrieved 52,046 patents and 1,801,075 publications. The top performing technology cluster of the last 30 years was minimally invasive surgery. Robotic surgery, surgical staplers, and image guidance were the most emergent technology clusters. When examining the growth curves for these clusters they were found to follow an S-shaped pattern of growth, with the emergent technologies lying on the exponential phases of their respective growth curves. In addition, publication and patent counts were closely correlated in areas of technology expansion. This article demonstrates the utility of publically available patent and publication data to quantify innovations within surgical technology and proposes a novel methodology for assessing and forecasting areas of technological innovation.

  14. 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

  15. 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.

  16. Mammalian cloning: advances and limitations.

    PubMed

    Solter, D

    2000-12-01

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

  17. 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.

  18. 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.

  19. Cloning whole bacterial genomes in yeast

    PubMed Central

    Benders, Gwynedd A.; Noskov, Vladimir N.; Denisova, Evgeniya A.; Lartigue, Carole; Gibson, Daniel G.; Assad-Garcia, Nacyra; Chuang, Ray-Yuan; Carrera, William; Moodie, Monzia; Algire, Mikkel A.; Phan, Quang; Alperovich, Nina; Vashee, Sanjay; Merryman, Chuck; Venter, J. Craig; Smith, Hamilton O.; Glass, John I.; Hutchison, Clyde A.

    2010-01-01

    Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation. PMID:20211840

  20. Cloning whole bacterial genomes in yeast.

    PubMed

    Benders, Gwynedd A; Noskov, Vladimir N; Denisova, Evgeniya A; Lartigue, Carole; Gibson, Daniel G; Assad-Garcia, Nacyra; Chuang, Ray-Yuan; Carrera, William; Moodie, Monzia; Algire, Mikkel A; Phan, Quang; Alperovich, Nina; Vashee, Sanjay; Merryman, Chuck; Venter, J Craig; Smith, Hamilton O; Glass, John I; Hutchison, Clyde A

    2010-05-01

    Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation.

  1. 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

  2. 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)

  3. 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.

  4. 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)

  5. 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…

  6. 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…

  7. 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.

  8. Drawing from Freirian empowerment methods to develop and use innovative learning maps: increasing enrollment of uninsured children on Detroit's eastside.

    PubMed

    Lopez, Ellen D S; Lichtenstein, Richard; Lewis, Alonzo; Banaszak-Holl, Jane; Lewis, Cheryl; Johnson, Penni; Riley, Scherry; Baum, Nancy M

    2007-04-01

    In 2001, virtually every child on Detroit's eastside was eligible for health coverage, yet approximately 3,000 children remained uninsured. The primary aim of the Eastside Access Partnership (EAP), a community-based participatory research collaboration, was to increase enrollment of uninsured children in state programs. To achieve this aim, one of the approaches that EAP is using is the innovative Learning Map titled Choosing the Healthy Path, which was developed in collaboration with Root Learning, Inc. Although Learning Maps were originally developed to assist corporations in implementing strategic change, their integration of visualization and interactive dialogue incorporates Freirian principles of empowerment education, making them a viable option for providing meaningful learning opportunities for community residents. This article presents the collaborative process involving the University of Michigan, local community-based organizations, community members, and Root Learning consultants to develop a visual map that enables community residents to understand and overcome the barriers that prevent them from obtaining health insurance for their children.

  9. Autologous osteoblast transplantation, an innovative method of bone defect treatment: role of a tissue and cell bank in the process.

    PubMed

    Olender, E; Brubaker, S; Uhrynowska-Tyszkiewicz, I; Wojtowicz, A; Kaminski, A

    2014-10-01

    The idea of cell treatment of various diseases and medical conditions has become very popular. Some procedures are well established, as is autologous chondrocyte implantation, whereas others are still in the process of early development, laboratory experiments, and some clinical trials. This report is devoted to an example of an emerging cell treatment: bone augmentation with the use of autologous cells and its legal and technical background. Various requirements set by law must be met by tissue banks performing cell seeding of grafts. In Europe, the requirements are described in directives 2004/23/EC, 2006/17/EC, 2006/86/EC, and in the regulation 2007/1394/EC. Revitalization of biostatic allografts gives new, promising tools for creation of functional parts of organs; brings the methodology used in tissue banks closer to tissue engineering; places the enterprise in the mainstream of advanced biotechnology; allows the full potential of tissue allografts; and opens a new, large area for clinical and laboratory research. Cell and tissue processing also have a financial impact on the treatment: it produces additional expenditures. Clinical effectiveness will be the most decisive factor of whether this innovative treatment will be applied in a particular type of medical condition. From a tissue establishment perspective, the most important issue is to develop a procedure that ensures safety for the patient in graft quality terms.

  10. Cloning of fish enzymes and other fish protein genes.

    PubMed

    Macouzet, M; Simpson, B K; Lee, B H

    1999-01-01

    Fish metabolism needs special enzymes that have maximum activity at very different conditions than their mammalian counterparts. Due to the differences in activity, these enzymes, especially cold-adapted proteases, could be used advantageously for the production of some foods. In addition to the enzymes, this review describes some other unique fish polypeptides such as antifreeze proteins, fluorescent proteins, antitumor peptides, antibiotics, and hormones, that have already been cloned and used in food processing, genetic engineering, medicine, and aquaculture. Recombinant DNA technology, which allows these biological molecules to be cloned and overexpressed in microorganisms is also described, highlighting innovative applications. The expected impact of cloning fish proteins in different fields of technology is discussed.

  11. 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.

  12. 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…

  13. 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.

  14. Innovation Expo

    NASA Image and Video Library

    2016-11-03

    Representatives from Parker Brothers, known for designing and building outrageous custom vehicles, spoke to employees at NASA's Kennedy Space Center in Florida about their creative thinking processes. The event took place during the center's 2016 Innovation Expo. Employees also were given an up-close look at their Neutron bike, featured in the movie “Tron.”

  15. Innovation Expo

    NASA Image and Video Library

    2016-11-03

    During the 2016 Innovation Expo at NASA's Kennedy Space Center in Florida, representatives from Parker Brothers, known for designing and building outrageous custom vehicles, spoke to Kennedy employees about their creative thinking processes. Employees also were given an up-close look at their Neutron bike, featured in the movie “Tron.”

  16. 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…

  17. 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.

  18. 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

  19. 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…

  20. Lessons learned from cloning dogs.

    PubMed

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

    2012-08-01

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

  1. 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.

  2. 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.

  3. Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material.

    PubMed

    De Backer, Benjamin; Debrus, Benjamin; Lebrun, Pierre; Theunis, Laetitia; Dubois, Nathalie; Decock, Lies; Verstraete, Alain; Hubert, Philippe; Charlier, Corinne

    2009-12-15

    GC is commonly used for the analysis of cannabis samples, e.g. in forensic chemistry. However, as this method is based on heating of the sample, acidic forms of cannabinoids are decarboxylated into their neutral counterparts. Conversely, HPLC permits the determination of the original composition of plant cannabinoids by direct analysis. Several HPLC methods have been described in the literature, but most of them failed to separate efficiently all the cannabinoids or were not validated according to general guidelines. By use of an innovative methodology for modelling chromatographic responses, a simple and accurate HPLC/DAD method was developed for the quantification of major neutral and acidic cannabinoids present in cannabis plant material: Delta9-tetrahydrocannabinol (THC), THC acid (THCA), cannabidiol (CBD), CBD acid (CBDA), cannabigerol (CBG), CBG acid (CBGA) and cannabinol (CBN). Delta8-Tetrahydrocannabinol (Delta8-THC) was determined qualitatively. Following the practice of design of experiments, predictive multilinear models were developed and used in order to find optimal chromatographic analytical conditions. The method was validated following an approach using accuracy profiles based on beta-expectation tolerance intervals for the total error measurement, and assessing the measurements uncertainty. This analytical method can be used for diverse applications, e.g. plant phenotype determination, evaluation of psychoactive potency and control of material quality.

  4. Positional cloning by linkage disequilibrium.

    PubMed

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

    2004-05-01

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

  5. 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…

  6. 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…

  7. Equine cloning: applications and outcomes.

    PubMed

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

    2006-01-01

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

  8. Membrane Innovation in Dialysis.

    PubMed

    Boschetti-de-Fierro, Adriana; Beck, Werner; Hildwein, Helmut; Krause, Bernd; Storr, Markus; Zweigart, Carina

    2017-01-01

    Despite advances in renal replacement therapy, the adequate removal of uremic toxins over a broad molecular weight range remains one of the unmet needs in hemodialysis. Therefore, membrane innovation is currently directed towards enhanced removal of uremic toxins and increased membrane permeability. This chapter presents a variety of opportunities where innovation is brought into dialysis membranes. It covers the membrane formation from solution, describing different approaches to control the phase inversion process through additives that either swell in the polymer solution or influence the pore shrinkage during the membrane drying process. Additionally, large-scale manufacturing is described, and the influence of raw materials, spinning, and drying processes on membrane selectivity are presented. Finally, new characterization methods developed for the latest innovations around the application of membranes in dialysis are discussed, which allow the membrane performance for removal of a broad range of uremic toxins and the expected albumin loss in clinical use. © 2017 S. Karger AG, Basel.

  9. 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.

  10. 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.

  11. 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

  12. Positional cloning in maize (Zea mays subsp. mays, Poaceae)1

    PubMed Central

    Gallavotti, Andrea; Whipple, Clinton J.

    2015-01-01

    • Premise of the study: Positional (or map-based) cloning is a common approach to identify the molecular lesions causing mutant phenotypes. Despite its large and complex genome, positional cloning has been recently shown to be feasible in maize, opening up a diverse collection of mutants to molecular characterization. • Methods and Results: Here we outline a general protocol for positional cloning in maize. While the general strategy is similar to that used in other plant species, we focus on the unique resources and approaches that should be considered when applied to maize mutants. • Conclusions: Positional cloning approaches are appropriate for maize mutants and quantitative traits, opening up to molecular characterization the large array of genetic diversity in this agronomically important species. The cloning approach described should be broadly applicable to other species as more plant genomes become available. PMID:25606355

  13. The development and use of an innovative laboratory method for measuring arsenic in drinking water from western Bangladesh.

    PubMed

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

    2005-09-01

    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 microg/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 microg/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 microg/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 microg/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 microg/L.

  14. 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

  15. Application of an innovative design space optimization strategy to the development of LC methods for the simultaneous screening of antibiotics to combat poor quality medicines.

    PubMed

    Mbinze, J K; Dispas, A; Lebrun, P; Mavar Tayey Mbay, J; Habyalimana, V; Kalenda, N; Rozet, E; Hubert, Ph; Marini, R D

    2013-11-01

    The poor quality of medicines is a crucial problem of public health. Therefore, it is important to have analytical tools to attend decisions of the legal authorities while combating this offense. In this context, the main objective of this study was to develop generic methods able to trace, screen and determine several antibiotics and common associated molecules by mean of liquid chromatographic techniques. For that purpose, an innovative Design Space optimization strategy was applied, targeting 16 antibiotics and 3 beta-lactamase inhibitors. The robustness of the developed method allowed using its use in an environment where operational factors such as temperature are not easy to control and eased its transfer to Ultra High Performance Liquid Chromatography. To demonstrate its ability to quantify the targeted molecules, the developed and transferred method was fully validated for two active ingredients commonly used in association, sulbactam and ceftriaxone, using the accuracy profile as decision tool. Based on this successful step, the method was then used for the quantitative determination of these two active ingredients in three pharmaceutical brands marketed in the Democratic Republic of Congo. Two out of the three pharmaceutical products did not comply with the specifications.

  16. Delivery Innovations.

    PubMed

    2017-03-01

    The need for innovations in care delivery is recognized by providers, payers, and patients alike. Hospitals, physicians, and other clinicians are experimenting with new models of care designed to better meet patients' needs, reduce administrative burdens, and lower costs. The Affordable Care Act placed the Medicare and Medicaid programs at the center of a national effort to experiment with delivery and payment models designed to improve care and contain costs. These public-sector efforts have often aligned with private initiatives, such as the use of reference pricing-in which an insurer will only pay for a service at the price available from the lowest-cost provider. Employers in the public and private sectors have adopted value-based insurance design, in which copayments and deductibles are calibrated to the clinical benefit obtained from different services. Patients have the most to gain-or lose-from delivery innovations. Better, more efficient care should translate into better health and lower costs, but payment models designed to encourage innovation may have the unintended effect of limiting access to care.

  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. An innovative method to acquire the location of point A for cervical cancer treatment by HDR brachytherapy.

    PubMed

    Chang, Liyun; Ho, Sheng-Yow; Yeh, Shyh-An; Lee, Tsair-Fwu; Chen, Pang-Yu

    2016-11-01

    Brachytherapy of local cervical cancer is generally accomplished through film-based treatment planning with the prescription directed to point A, which is invisible on images and is located at a high-dose gradient area. Through a standard reconstruction method by digitizing film points, the location error for point A would be 3 mm with a condition of 30° curvature tandem, which is 10° away from the gantry rotation axis of a simulator, and has an 8.7 cm interval between the flange and the isocenter. To reduce the location error of the reconstructed point A, this paper proposes a method and demonstrates its accuracy. The Cartesian coordinates of point A were derived by acquiring the locations of the cervical os (tandem flange) and a dummy seed located in the tandem above the flange. To verify this analytical method, ball marks in a commercial "Isocentric Beam Checker" were selected to simulate the two points A, the os, and the dummies. The Checker was placed on the simulator couch with its center ball coincident with the simulator isocenter and its rotation axis perpendicular to the gantry rotation axis. With different combinations of the Checker and couch rotation angles, the orthogonal films were shot and all coordinates of the selected points were reconstructed through the treatment planning system and compared with that calculated through the analytical method. The position uncertainty and the deviation prediction of point A were also evaluated. With a good choice of the reference dummy point, the position deviations of point A obtained through this analytical method were found to be generally within 1 mm, with the standard uncertainty less than 0.5 mm. In summary, this new method is a practical and accurate tool for clinical usage to acquire the accurate location of point A for the treatment of cervical cancer patient. PACS number(s): 87.55.km. © 2016 The Authors.

  20. An innovative method to acquire the location of point A for cervical cancer treatment by HDR brachytherapy.

    PubMed

    Chang, Liyun; Ho, Sheng-Yow; Yeh, Shyh-An; Lee, Tsair-Fwu; Chen, Pang-Yu

    2016-11-08

    Brachytherapy of local cervical cancer is generally accomplished through film-based treatment planning with the prescription directed to point A, which is invisible on images and is located at a high-dose gradient area. Through a standard reconstruction method by digitizing film points, the location error for point A would be 3mm with a condition of 30° curvature tandem, which is 10° away from the gantry rotation axis of a simulator, and has an 8.7 cm interval between the flange and the isocenter. To reduce the location error of the reconstructed point A, this paper proposes a method and demonstrates its accuracy. The Cartesian coordinates of point A were derived by acquiring the locations of the cervical os (tandem flange) and a dummy seed located in the tandem above the flange. To verify this analytical method, ball marks in a commercial "Isocentric Beam Checker" were selected to simulate the two points A, the os, and the dummies. The Checker was placed on the simulator couch with its center ball coincident with the simulator isocenter and its rotation axis perpendicular to the gantry rotation axis. With different combinations of the Checker and couch rotation angles, the orthogonal films were shot and all coor-dinates of the selected points were reconstructed through the treatment planning system and compared with that calculated through the analytical method. The position uncertainty and the deviation prediction of point A were also evaluated. With a good choice of the reference dummy point, the position deviations of point A obtained through this analytical method were found to be generally within 1 mm, with the standard uncertainty less than 0.5 mm. In summary, this new method is a practical and accurate tool for clinical usage to acquire the accurate location of point A for the treatment of cervical cancer patient.