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Sample records for raising cre strategic

  1. Raising financing through strategic timing

    NASA Astrophysics Data System (ADS)

    Maine, Elicia; Thomas, V. J.

    2017-02-01

    Strategic timing can be key for nano-drug-delivery ventures to get financing. Timely publications engage potential partners; early broad, blocking, relevant patents demonstrate the potential to appropriate value; and venture formation closer to clinical viability better aligns its timeline with that of venture capitalists.

  2. Essential Not Optional: A Strategic Approach to Fund-Raising for Endowments

    ERIC Educational Resources Information Center

    Griswold, John S.; Jarvis, William F.

    2012-01-01

    This overview of the state of endowment giving comes at a time of urgent need in the nonprofit sector. With market returns uncertain, and spending restraint difficult, the moderate but measurable increase in donations in the previous year invites institutions to consider elevating fund-raising to a more strategic position within the organization.…

  3. Strategic Planning, Marketing & Public Relations, and Fund-Raising in Higher Education: Perspectives, Readings, and Annotated Bibliography.

    ERIC Educational Resources Information Center

    Ryans, Cynthia C.; Shanklin, William L.

    Perspectives on college and university strategic planning, marketing and public relations, and fund-raising are offered. Also included are previously published journal articles by experts in this area, annotated bibliographies of books and journal articles on these subjects, author/title and subject indexes, and a directory of publishers. Three…

  4. [The WHO age-friendly cities program raises the issue of strategic planning, coordination and local political structure].

    PubMed

    Chapon, Pierre-Marie

    2011-06-01

    In 2005 the World Health Organization initiated the «global age-friendly cities project» which encourages cities to be more inclusive of older people and to develop seniors' involvement by setting up a continuous cycle of assessment and improvement of urban living. The conclusions of the first french audit, which was made in Lyon in 2011, raise the issue of the capacity of cities to implement projects in all the areas fostering active aging. The question of strategic planning and of the level of intervention is relevant for the topics selected, and especially that of health. Is the present organization between the different actors satisfactory, or should large cities have more power? Co-ordination is necessary, and the creation of «gerontopoles» could bring an answer.

  5. Safety assessment of cre recombinase.

    PubMed

    Hileman, Ronald E; Bonner, Heather K S; Kaempfe, Terry A; Hammond, Bruce G; Glenn, Kevin C

    2006-11-01

    Cre recombinase, when used as a tool in agricultural biotechnology, can precisely excise DNA sequences that may be useful in the introduction of a new trait but are not needed in the commercial product. Although the cre genetic material would not be present in the final product, the present studies were performed to assess the safety of Cre recombinase to provide confirmatory evidence of the safe use of Cre-lox technology in agricultural biotechnology. Cre recombinase shares no relevant sequence similarity to known allergens or toxins. When Cre recombinase was exposed to a pH 1.2 solution of simulated gastric fluid lacking pepsin, CD spectroscopy showed that there was a loss of secondary structure and that the protein was no longer active in a functional assay. Cre recombinase was degraded rapidly when exposed to pepsin in a standardized gastric digestion model; therefore, Cre recombinase would not survive the harsh gastric environment. When orally administered to mice as an acute dosage of 53 mg/kg of body weight, no treatment-related adverse findings were observed. These data support the conclusion that human and animal dietary exposure to Cre recombinase pose no known safety concerns; consistent with the fact that bacteriophage P1, the source of the cre gene and expressed protein, is commonly encountered in the environment and in normal enteric bacteria without reports of adverse consequences.

  6. Foxn1[Cre] Expression in the Male Germline

    PubMed Central

    Shi, Jianjun; Getun, Irina; Torres, Bivian; Petrie, Howard T.

    2016-01-01

    Foxn1 (forkhead box N1), also known as the nude gene or winged-helix nude (Whn), is a forkhead transcription factor thought to be restricted to keratinocytes in the skin and thymus. Consistent with this tissue distribution, spontaneous or targeted mutation of Foxn1 results in the absence of both hair and a thymus. Genetic manipulation of the Foxn1 locus thus represents a powerful tool for tissue specific gene control in the skin and thymus, and tools such as Cre recombinase under control of the Foxn1 locus are widely used for this purpose. Unexpectedly, we show that Foxn1[Cre] exhibits unexpected activity in male germ cells, resulting in ubiquitous targeting of loxP-flanked alleles in all tissues in offspring from Foxn1[Cre] expressing male mice. Inheritance of recombined loxP alleles occurs independently of Cre inheritance (i.e., offspring lacking Cre nonetheless exhibit recombined alleles), suggesting that Foxn1[Cre] induced recombination in male germ cells must occur prior to meiosis in diploid germ cells. Together with previously published data, our results show that Foxn1, and alleles under its control, are expressed in the pre-meiotic male germline, revealing a new tool for germline targeting of genes, and raising important concerns for gender selection when using Foxn1 regulatory elements. PMID:27880796

  7. Unmodified Cre recombinase crosses the membrane

    PubMed Central

    Will, Elke; Klump, Hannes; Heffner, Nicole; Schwieger, Maike; Schiedlmeier, Bernhard; Ostertag, Wolfram; Baum, Christopher; Stocking, Carol

    2002-01-01

    Site-specific recombination in genetically modified cells can be achieved by the activity of Cre recombinase from bacteriophage P1. Commonly an expression vector encoding Cre is introduced into cells; however, this can lead to undesired side-effects. Therefore, we tested whether cell-permeable Cre fusion proteins can be directly used for lox-specific recombination in a cell line tailored to shift from red to green fluorescence after loxP-specific recombination. Comparison of purified recombinant Cre proteins with and without a heterologous ‘protein transduction domain’ surprisingly showed that the unmodified Cre recombinase already possesses an intrinsic ability to cross the membrane border. Addition of purified recombinant Cre enyzme to primary bone marrow cells isolated from transgenic C/EBPαfl/fl mice also led to excision of the ‘floxed’ C/EBPα gene, thus demonstrating its potential for in vivo applications. We conclude that Cre enyzme itself or its intrinsic membrane-permeating moiety are attractive tools for direct manipulation of mammalian cells. PMID:12060697

  8. Skeletal defects in Osterix-Cre transgenic mice.

    PubMed

    Huang, Wei; Olsen, Bjorn R

    2015-02-01

    Cre/loxP recombination is a powerful strategy widely used for in vivo conditional gene targeting. This technique has made possible many important discoveries of gene function in normal and disease biology. However, due to the transgenic nature of most Cre mouse strains undesired phenotypes occasionally occur in Cre mice. Here we report skeletal defects in Osterix-Cre (Osx-Cre) transgenic mice including delayed calvarial ossification and fracture calluses at multiple skeletal sites. These data suggest that Osx-Cre containing controls should be used for both in vivo and in vitro skeletal analyses of conditional knockout mice generated with this Osx-Cre mouse strain.

  9. Raising consciousness.

    PubMed

    Hirsch, Joy

    2005-05-01

    The national debate over Terri Schiavo exposed a critical gap between emotional fervor about brain-injured patients and the medical science that informs standards of care for them. Some of the questions raised in the public and legal forums point to a need for research and enhanced understanding of the mechanisms of recovery from disorders of consciousness.

  10. Raising Feminists.

    ERIC Educational Resources Information Center

    Katz, Phyllis A.

    1996-01-01

    Considers reasons for raising feminist children, how this concept might be operationalized, and what the correlates of such gender-flexible patterns might be. Results from longitudinal data involving 200 children suggest that variation exists in the degree to which even young children subscribe to stereotypes. These variations seem to relate to…

  11. Beyond knockouts: cre resources for conditional mutagenesis

    PubMed Central

    Murray, Stephen A.; Eppig, Janan T.; Smedley, Damian; Simpson, Elizabeth M.; Rosenthal, Nadia

    2013-01-01

    With the effort of the International Phenotyping Consortium (IMPC) to produce thousands of strains with conditional potential gathering steam, there is growing recognition that it must be supported by a rich toolbox of cre driver strains. The approaches to build cre strains have evolved in both sophistication and reliability, replacing first generation strains with tools that can target individual cell populations with incredible precision and specificity. The modest set of cre drivers generated by individual labs over the past 15+ years is now growing rapidly, thanks to a number of large-scale projects to produce new cre strains for the community. The power of this growing resource, however, depends upon the proper deep characterization of strain function, as even the best designed strain can display a variety of undesirable features that must be considered in experimental design. This must be coupled with the parallel development of informatics tools to provide functional data to the user, and facilitated access to the strains through public repositories. We will discuss the current progress on all of these fronts and the challenges that remain to ensure the scientific community can capitalize on the tremendous number of mouse resources at their disposal. PMID:22926223

  12. Photo-activatable Cre recombinase regulates gene expression in vivo.

    PubMed

    Schindler, Suzanne E; McCall, Jordan G; Yan, Ping; Hyrc, Krzystof L; Li, Mingjie; Tucker, Chandra L; Lee, Jin-Moo; Bruchas, Michael R; Diamond, Marc I

    2015-09-09

    Techniques allowing precise spatial and temporal control of gene expression in the brain are needed. Herein we describe optogenetic approaches using a photo-activatable Cre recombinase (PA-Cre) to stably modify gene expression in the mouse brain. Blue light illumination for 12 hours via optical fibers activated PA-Cre in the hippocampus, a deep brain structure. Two-photon illumination through a thinned skull window for 100 minutes activated PA-Cre within a sub-millimeter region of cortex. Light activation of PA-Cre may allow permanent gene modification with improved spatiotemporal precision compared to standard methods.

  13. CreZOO—the European virtual repository of Cre and other targeted conditional driver strains

    PubMed Central

    Chandras, Christina; Zouberakis, Michael; Salimova, Ekaterina; Smedley, Damian; Rosenthal, Nadia; Aidinis, Vassilis

    2012-01-01

    The CreZOO (http://www.crezoo.org/) is the European virtual repository of Cre and other targeted conditional driver strains. These mice serve as tools for researchers to selectively ‘switch off’ gene expression in mouse models to examine gene function and disease pathology. CreZOO aims to capture and disseminate extant and new information on these Cre driver strains, such as genetic background and availability information, and details pertaining promoter, allele, inducibility and expression patterns, which are also presented. All transgenic strains carry detailed information according to MGI's official nomenclature, whereas their availability [e.g. live mice, cryopreserved embryos, sperm and embryonic stem (ES) cells] is clearly indicated with links to European and International databases and repositories (EMMA, MGI/IMSR, MMRRC, etc) and laboratories where the particular mouse strain is available together with the respective IDs. Each promoter/gene includes IDs and direct links to MGI, Entrez Gene, Ensembl, OMIM and RGD databases depending on their species origin, whereas allele information is presented with MGI IDs and active hyperlinks to redirect the user to the respective page in a new tab. The tissue/cell (special) and developmental (temporal) specificity expression patterns are clearly presented, whereas handling and genotyping details (in the form of documents or hyperlinks) together with all relevant publications are clearly presented with PMID(s) and direct PubMed links. CreZOO's design offers a user-friendly query interface and provides instant access to the list of conditional driver strains, promoters and inducibility details. Database access is free of charge and there are no registration requirements for data querying. CreZOO is being developed in the context of the CREATE consortium (http://www.creline.org/), a core of major European and international mouse database holders and research groups involved in conditional mutagenesis. Database URL

  14. A Study of Plazomicin Compared With Colistin in Patients With Infection Due to Carbapenem-Resistant Enterobacteriaceae (CRE)

    ClinicalTrials.gov

    2016-10-03

    Bloodstream Infections (BSI) Due to CRE; Hospital-Acquired Bacterial Pneumonia (HABP) Due to CRE; Ventilator-Associated Bacterial Pneumonia (VABP) Due to CRE; Complicated Urinary Tract Infection (cUTI) Due to CRE; Acute Pyelonephritis (AP) Due to CRE

  15. Inducible control of gene expression with destabilized Cre

    PubMed Central

    Sando, Richard; Baumgaertel, Karsten; Pieraut, Simon; Torabi-Rander, Nina; Wandless, Thomas J.; Mayford, Mark; Maximov, Anton

    2014-01-01

    Acute manipulation of gene and protein function in the brain is essential for understanding the mechanisms of nervous system development, plasticity and information processing. Here we describe a technique based on a destabilized Cre recombinase (DD-Cre) whose activity is controlled by the antibiotic, TMP. We show that DD-Cre triggers rapid TMP-dependent recombination of “floxed” alleles in mouse neurons in vivo, and validate the use of this system for neurobehavioral research. PMID:24056874

  16. Identification of the CRE-1 Cellulolytic Regulon in Neurospora crassa

    PubMed Central

    Sun, Jianping; Glass, N. Louise

    2011-01-01

    Background In filamentous ascomycete fungi, the utilization of alternate carbon sources is influenced by the zinc finger transcription factor CreA/CRE-1, which encodes a carbon catabolite repressor protein homologous to Mig1 from Saccharomyces cerevisiae. In Neurospora crassa, deletion of cre-1 results in increased secretion of amylase and β-galactosidase. Methodology/Principal Findings Here we show that a strain carrying a deletion of cre-1 has increased cellulolytic activity and increased expression of cellulolytic genes during growth on crystalline cellulose (Avicel). Constitutive expression of cre-1 complements the phenotype of a N. crassa Δcre-1 strain grown on Avicel, and also results in stronger repression of cellulolytic protein secretion and enzyme activity. We determined the CRE-1 regulon by investigating the secretome and transcriptome of a Δcre-1 strain as compared to wild type when grown on Avicel versus minimal medium. Chromatin immunoprecipitation-PCR of putative target genes showed that CRE-1 binds to only some adjacent 5′-SYGGRG-3′ motifs, consistent with previous findings in other fungi, and suggests that unidentified additional regulatory factors affect CRE-1 binding to promoter regions. Characterization of 30 mutants containing deletions in genes whose expression level increased in a Δcre-1 strain under cellulolytic conditions identified novel genes that affect cellulase activity and protein secretion. Conclusions/Significance Our data provide comprehensive information on the CRE-1 regulon in N. crassa and contribute to deciphering the global role of carbon catabolite repression in filamentous ascomycete fungi during plant cell wall deconstruction. PMID:21980519

  17. Phenotypically silent Cre recombination within the postnatal ventricular conduction system

    PubMed Central

    Bhattacharyya, Samadrita; Bhakta, Minoti

    2017-01-01

    The cardiac conduction system (CCS) is composed of specialized cardiomyocytes that initiate and maintain cardiac rhythm. Any perturbation to the normal sequence of electrical events within the heart can result in cardiac arrhythmias. To understand how cardiac rhythm is established at the molecular level, several genetically modified mouse lines expressing Cre recombinase within specific CCS compartments have been created. In general, Cre driver lines have been generated either by homologous recombination of Cre into an endogenous locus or Cre expression driven by a randomly inserted transgene. However, haploinsufficiency of the endogenous gene compromises the former approach, while position effects negatively impact the latter. To address these limitations, we generated a Cre driver line for the ventricular conduction system (VCS) that preserves endogenous gene expression by targeting the Contactin2 (Cntn2) 3’ untranslated region (3’UTR). Here we show that Cntn23’UTR-IRES-Cre-EGFP/+ mice recombine floxed alleles within the VCS and that Cre expression faithfully recapitulates the spatial distribution of Cntn2 within the heart. We further demonstrate that Cre expression initiates after birth with preservation of native Cntn2 protein. Finally, we show that Cntn23’UTR-IRES-Cre-EGFP/+ mice maintain normal cardiac mechanical and electrical function. Taken together, our results establish a novel VCS-specific Cre driver line without the adverse consequences of haploinsufficiency or position effects. We expect that our new mouse line will add to the accumulating toolkit of CCS-specific mouse reagents and aid characterization of the cell-autonomous molecular circuitry that drives VCS maintenance and function. PMID:28358866

  18. Germ-Line Recombination Activity of the Widely Used hGFAP-Cre and Nestin-Cre Transgenes

    PubMed Central

    Zhang, Jiong; Dublin, Pavel; Griemsmann, Stephanie; Klein, Alexandra; Brehm, Ralph; Bedner, Peter; Fleischmann, Bernd K.; Steinhäuser, Christian; Theis, Martin

    2013-01-01

    Herein we demonstrate with PCR, immunodetection and reporter gene approaches that the widely used human Glial Fibrillary Acidic Protein (hGFAP)-Cre transgene exhibits spontaneous germ-line recombination activity in leading to deletion in brain, heart and tail tissue with high frequency. The ectopic activity of hGFAP-Cre requires a rigorous control. We likewise observed that a second widely used nestin-Cre transgene shows germ-line deletion. Here we describe procedures to identify mice with germ-line recombination mediated by the hGFAP-Cre and nestin-Cre transgenes. Such control is essential to avoid pleiotropic effects due to germ-line deletion of loxP-flanked target genes and to maintain the CNS-restricted deletion status in transgenic mouse colonies. PMID:24349371

  19. Gene deletion in urothelium by specific expression of Cre recombinase.

    PubMed

    Mo, Lan; Cheng, Jin; Lee, Eva Y-H P; Sun, Tung-Tien; Wu, Xue-Ru

    2005-09-01

    Urothelium that lines almost the entire urinary tract acts as a permeability barrier and is involved in the pathogenesis of major urinary diseases, including urothelial carcinoma, urinary tract infection, and interstitial cystitis. However, investigation of urothelial biology and diseases has been hampered by the lack of tissue-specific approaches. To address this deficiency, we sought to develop a urothelium-specific knockout system using the Cre/loxP strategy. Transgenic mouse lines were generated in which a 3.6-kb mouse uroplakin II (UPII) promoter was used to drive the expression of Cre recombinase (Cre). Among the multiple tissues analyzed, Cre was found to be expressed exclusively in the urothelia of the transgenic mice. Crossing a UPII-Cre transgenic line with a ROSA26-LacZ reporter line, in which LacZ expression depends on Cre-mediated deletion of a floxed "stop" sequence, led to LacZ expression only in the urothelium. Gene recombination was also observed when the UPII-Cre line was crossed to an independent line in which a part of the p53 gene was flanked by the loxP sequences (floxed p53). Truncation of the p53 gene and mRNA was observed exclusively in the urothelia of double transgenic mice harboring both the UPII-Cre transgene and the floxed p53 allele. These results demonstrate for the first time the feasibility and potentially wide applicability of the UPII-Cre transgenic mice to inactivate any genes of interest in the urothelium.

  20. Raise Your Profile: Build Your Program

    ERIC Educational Resources Information Center

    Nutefall, Jennifer E.; Gaspar, Deborah

    2008-01-01

    To raise the library's profile within the campus community, it is critical to create a strategic plan and align library goals with those of the university. At George Washington University's Gelman Library, the instruction librarians gained internal and external support to hire two new instruction librarians to better support collaboration with the…

  1. Generation of AQP2-Cre transgenic mini-pigs specifically expressing Cre recombinase in kidney collecting duct cells.

    PubMed

    Luo, Weiwei; Li, Zhanjun; Huang, Yongye; Han, Yang; Yao, Chaogang; Duan, Xinping; Ouyang, Hongsheng; Li, Li

    2014-04-01

    The important differences in physiological parameters and anatomical characteristics of the kidney between humans and mice make it difficult to replicate the precise progression of human renal cystic diseases in gene modification mouse models. In contrast to mice, pigs are a better animal model of human diseases, as they are more similar in terms of organ size, structure, and physiological parameters. Here, we report the generation and initial examination of an AQP2-Cre transgenic (Tg) Chinese miniature (mini)-pig line that expresses Cre recombinase exclusively in kidney collecting duct cells. An 8-kb fragment of the mini-pig aquaporin 2 (AQP2) 5'-flanking region was utilized to direct Cre expression in Tg mini-pigs. Two Tg mini-pigs were generated by pig somatic cell nuclear transfer and both carried the entire coding sequence of Cre recombinase. RT-PCR and western blotting analysis revealed that Cre recombinase was uniquely expressed in the kidney, while immunohistochemical studies located its expression in kidney collecting duct cells. Furthermore, six integration sites and 12-14 copies of the Cre gene were detected in various tissues by high-efficiency thermal asymmetric interlaced PCR and absolute quantitative real-time PCR, respectively. Combined with previous studies of Cre recombinase activity, we believe that this AQP2-Cre Tg mini-pig line will be a useful tool to generate kidney collecting duct cell-specific gene knockout mini-pig models, thereby allowing the investigation of gene functions in kidney development and the mechanisms of human renal cystic disease.

  2. Tissue Specific Expression of Cre in Rat Tyrosine Hydroxylase and Dopamine Active Transporter-Positive Neurons

    PubMed Central

    Liu, Zhenyi; Brown, Andrew; Fisher, Dan; Wu, Yumei; Warren, Joe; Cui, Xiaoxia

    2016-01-01

    The rat is a preferred model system over the mouse for neurological studies, and cell type-specific Cre expression in the rat enables precise ablation of gene function in neurons of interest, which is especially valuable for neurodegenerative disease modeling and optogenetics. Yet, few such Cre rats are available. Here we report the characterization of two Cre rats, tyrosine hydroxylase (TH)-Cre and dopamine active transporter (DAT or Slc6a3)-Cre, by using a combination of immunohistochemistry (IHC) and mRNA fluorescence in situ hybridization (FISH) as well as a fluorescent reporter for Cre activity. We detected Cre expression in expected neurons in both Cre lines. Interestingly, we also found that in Th-Cre rats, but not DAT-Cre rats, Cre is expressed in female germ cells, allowing germline excision of the floxed allele and hence the generation of whole-body knockout rats. In summary, our data demonstrate that targeted integration of Cre cassette lead to faithful recapitulation of expression pattern of the endogenous promoter, and mRNA FISH, in addition to IHC, is an effective method for the analysis of the spatiotemporal gene expression patterns in the rat brain, alleviating the dependence on high quality antibodies that are often not available against rat proteins. The Th-Cre and the DAT-Cre rat lines express Cre in selective subsets of dopaminergic neurons and should be particularly useful for researches on Parkinson’s disease. PMID:26886559

  3. Thinking Strategically.

    ERIC Educational Resources Information Center

    Jeffress, Conway

    2000-01-01

    Asserts that community college leaders must think strategically and understand the difference between what is important and immediate, and what is strategic and essential to the long-term survival of a college. States that thinking strategically aligns decision-making and actions with the core purpose of the college; produces core competencies in…

  4. Infection control implications of heterogeneous resistance mechanisms in carbapenem-resistant Enterobacteriaceae (CRE).

    PubMed

    Goodman, K E; Simner, P J; Tamma, P D; Milstone, A M

    2016-01-01

    The Centers for Disease Control and Prevention (CDC) defines carbapenem-resistant Enterobacteriaceae (CRE) based upon a phenotypic demonstration of carbapenem resistance. However, considerable heterogeneity exists within this definitional umbrella. CRE may mechanistically differ by whether they do or do not produce carbapenemases. Moreover, patients can acquire CRE through multiple pathways: endogenously through antibiotic selective pressure on intestinal microbiota, exogenously through horizontal transmission or through a combination of these factors. Some evidence suggests that non-carbapenemase-producing CRE may be more frequently acquired by antibiotic exposure and carbapenemase-producing CRE via horizontal transmission, but definitive data are lacking. This review examines types of CRE resistance mechanisms, antibiotic exposure and horizontal transmission pathways of CRE acquisition, and the implications of these heterogeneities to the development of evidence-based CRE healthcare epidemiology policies. In our Expert Commentary & Five-Year View, we outline specific nosocomial CRE knowledge gaps and potential methodological approaches for their resolution.

  5. Heterogeneous transgene expression in the retinas of the TH-RFP, TH-Cre, TH-BAC-Cre and DAT-Cre mouse lines

    PubMed Central

    Vuong, Helen E.; de Sevilla Müller, Luis Pérez; Hardi, Claudia N.; McMahon, Douglas G.; Brecha, Nicholas C.

    2015-01-01

    Transgenic mouse lines are essential tools for understanding the connectivity, physiology and function of neuronal circuits, including those in the retina. This report compares transgene expression in the retina of a tyrosine hydroxylase (TH)-red fluorescent protein (RFP) line with three catecholamine-related Cre recombinase lines [TH-bacterial artificial chromosome (BAC)-, TH-, and dopamine transporter (DAT)-Cre] that were crossed with a ROSA26-tdTomato reporter line. Retinas were evaluated and immunostained with commonly used antibodies including those directed to TH, GABA and glycine to characterize the RFP or tdTomato fluorescent-labeled amacrine cells, and an antibody directed to RNA-binding protein with multiple splicing to identify ganglion cells. In TH-RFP retinas, types 1 and 2 dopamine (DA) amacrine cells were identified by their characteristic cellular morphology and type 1 DA cells by their expression of TH immunoreactivity. In the TH-BAC-, TH-, and DAT-tdTomato retinas, less than 1%, ~6%, and 0%, respectively, of the fluorescent cells were the expected type 1 DA amacrine cells. Instead, in the TH-BAC-tdTomato retinas, fluorescently labeled AII amacrine cells were predominant, with some medium somal diameter ganglion cells. In TH-tdTomato retinas, fluorescence was in multiple neurochemical amacrine cell types, including four types of polyaxonal amacrine cells. In DAT-tdTomato retinas, fluorescence was in GABA immunoreactive amacrine cells, including two types of bistratified and two types of monostratified amacrine cells. Although each of the Cre lines were generated with the intent to specifically label DA cells, our findings show a cellular diversity in Cre expression in the adult retina and indicate the importance of careful characterization of transgene labeling patterns. These mouse lines with their distinctive cellular labeling patterns will be useful tools for future studies of retinal function and visual processing. PMID:26335381

  6. Growth inhibition and DNA damage induced by Cre recombinase in mammalian cells

    PubMed Central

    Loonstra, Ate; Vooijs, Marc; Beverloo, H. Berna; Allak, Bushra Al; van Drunen, Ellen; Kanaar, Roland; Berns, Anton; Jonkers, Jos

    2001-01-01

    The use of Cre/loxP recombination in mammalian cells has expanded rapidly. We describe here that Cre expression in cultured mammalian cells may result in a markedly reduced proliferation and that this effect is dependent on the endonuclease activity of Cre. Chromosome analysis after Cre expression revealed numerous chromosomal aberrations and an increased number of sister chromatid exchanges. Titration experiments in mouse embryo fibroblasts with a ligand-regulatable Cre-ERT show that toxicity is dependent on the level of Cre activity. Prolonged, low levels of Cre activity permit recombination without concomitant toxicity. This urges for a careful titration of Cre activity in conditional gene modification in mammalian cells. PMID:11481484

  7. Cre-mediated recombination in cell lineages that express the progesterone receptor.

    PubMed

    Soyal, Selma M; Mukherjee, Atish; Lee, Kevin Y-S; Li, Jie; Li, Huaiguang; DeMayo, Francesco J; Lydon, John P

    2005-02-01

    Using gene-targeting methods, a progesterone receptor Cre knockin (PR-Cre) mouse was generated in which Cre recombinase was inserted into exon 1 of the PR gene. The insertion positions the Cre gene downstream (and under the specific control) of the endogenous PR promoter. As for heterozygotes for the progesterone receptor knockout (PRKO) mutation, mice heterozygous for the Cre knockin insertion are phenotypically indistinguishable from wildtype. Crossing the PR-Cre with the ROSA26R reporter revealed that Cre excision activity is restricted to cells that express PR in progesterone-responsive tissues such as the uterus, ovary, oviduct, pituitary gland, and mammary gland. Initial characterization of the PR-Cre mouse underscores the utility of this model to precisely ablate floxed target genes specifically in cell lineages that express the PR. In the wider context of female reproductive tissue ontology, this model will be indispensable in tracing the developmental fate of cell lineages that descend from PR positive progenitors.

  8. Strategic weapons

    SciTech Connect

    Not Available

    1991-09-01

    This paper describes the Defense Department's process for formulating its strategic weapons targeting policy and translating that policy into a nuclear war plan-the Single Integrated Operational Plan. GAO provides information on the relationship between the strategic nuclear targeting process and the determination of requirements for nuclear weapons and related delivery systems, level of civilian oversight, and categories and types of targets. These strategic nuclear weapons systems, commonly known as the triad, include land-based intercontinental ballistic missiles, submarine-launched ballistic missiles, and strategic bombers armed with nuclear bombs and missiles.

  9. The effect of conditional inactivation of beta 1 integrins using twist 2 Cre, Osterix Cre and osteocalcin Cre lines on skeletal phenotype.

    PubMed

    Shekaran, Asha; Shoemaker, James T; Kavanaugh, Taylor E; Lin, Angela S; LaPlaca, Michelle C; Fan, Yuhong; Guldberg, Robert E; García, Andrés J

    2014-11-01

    Skeletal development and growth are complex processes regulated by multiple microenvironmental cues, including integrin-ECM interactions. The β1 sub-family of integrins is the largest integrin sub-family and constitutes the main integrin binding partners of collagen I, the major ECM component of bone. As complete β1 integrin knockout results in embryonic lethality, studies of β1 integrin function in vivo rely on tissue-specific gene deletions. While multiple in vitro studies indicate that β1 integrins are crucial regulators of osteogenesis and mineralization, in vivo osteoblast-specific perturbations of β1 integrins have resulted in mild and sometimes contradictory skeletal phenotypes. To further investigate the role of β1 integrins on skeletal phenotype, we used the Twist2-Cre, Osterix-Cre and osteocalcin-Cre lines to generate conditional β1 integrin deletions, where Cre is expressed primarily in mesenchymal condensation, pre-osteoblast, and mature osteoblast lineage cells respectively within these lines. Mice with Twist2-specific β1 integrin disruption were smaller, had impaired skeletal development, especially in the craniofacial and vertebral tissues at E19.5, and did not survive beyond birth. Osterix-specific β1 integrin deficiency resulted in viable mice which were normal at birth but displayed early defects in calvarial ossification, incisor eruption and growth as well as femoral bone mineral density, structure, and mechanical properties. Although these defects persisted into adulthood, they became milder with age. Finally, a lack of β1 integrins in mature osteoblasts and osteocytes resulted in minor alterations to femur structure but had no effect on mineral density, biomechanics or fracture healing. Taken together, our data indicate that β1 integrin expression in early mesenchymal condensations play an important role in skeletal ossification, while β1 integrin-ECM interactions in pre-osteoblast, odontoblast- and hypertrophic chondryocyte

  10. The Effect of Conditional Inactivation of Beta 1 Integrins using Twist 2 Cre, Osterix Cre and Osteocalcin Cre Lines on Skeletal Phenotype

    PubMed Central

    Shekaran, Asha; Shoemaker, James T.; Kavanaugh, Taylor E.; Lin, Angela S.; LaPlaca, Michelle C.; Fan, Yuhong; Guldberg, Robert E.; García, Andrés J.

    2014-01-01

    Skeletal development and growth are complex processes regulated by multiple microenvironmental cues, including integrin-ECM interactions. The β1 sub-family of integrins is the largest integrin sub-family and constitutes the main integrin binding partners of collagen I, the major ECM component of bone. As complete β1 integrin integrin knockout results in embryonic lethality, studies of β1 integrin function in vivo rely on tissue-specific gene deletions. While multiple in vitro studies indicate that β1 integrins are crucial regulators of osteogenesis and mineralization, in vivo osteoblast-specific perturbations of β1 integrins have resulted in mild and sometimes contradictory skeletal phenotypes. To further investigate the role of β1 integrins on skeletal phenotype, we used the Twist2-Cre, Osterix-Cre and Osteocalcin-Cre lines to generate conditional β1 integrin deletions, where cre is expressed primarily in mesenchymal condensation, pre-osteoblast, and mature osteoblast lineage cells respectively within these lines. Mice with Twist2-specific β1 integrin disruption were smaller, had impaired skeletal development, especially in the craniofacial and vertebral tissues at E19.5, and did not survive beyond birth. Osterix-specific β1 integrin deficiency resulted in viable mice which were normal at birth but displayed early defects in calvarial ossification, incisor eruption and growth as well as femoral bone mineral density, structure, and mechanical properties. Although these defects persisted into adulthood, they became milder with age. Finally, a lack of β1 integrins in mature osteoblasts and osteocytes resulted in minor alterations to femur structure but had no effect on mineral density, biomechanics or fracture healing. Taken together, our data indicate that β1 integrin expression in early mesenchymal condensations play an important role in skeletal ossification, while β1 integrin-ECM interactions in pre-osteoblast, odontoblast- and hypertrophic

  11. Raising Children Again

    MedlinePlus

    ... email address Submit Home > Healthy Aging > Caregiving Healthy Aging Raising children again Benefits available More information on ... financial issues. Connect with other organizations Administration on Aging, HHS American Association of Retired Persons (AARP) Caring ...

  12. Raising the Titanic.

    ERIC Educational Resources Information Center

    Baker, Romona

    1990-01-01

    Described is an activity in which groups of students investigate engineering principles by writing a feasibility study to raise the luxury liner, Titanic. The problem statement and directions, and suggestions for problem solutions are included. (CW)

  13. Fund Raising with Panache.

    ERIC Educational Resources Information Center

    Dedman, Robert

    1985-01-01

    The key to fund raising is the donor, and fund raisers should learn to cultivate potential donors, approach them with goals compatible with their own, supplement their thinking, get them involved, and swamp them with gratitude. (MSE)

  14. Unexpected severe consequences of Pikfyve deletion by aP2- or Aq-promoter-driven Cre expression for glucose homeostasis and mammary gland development.

    PubMed

    Ikonomov, Ognian C; Sbrissa, Diego; Delvecchio, Khortnal; Rillema, James A; Shisheva, Assia

    2016-06-01

    Systemic deficiency of PIKfyve, the evolutionarily conserved phosphoinositide kinase synthesizing cellular PtdIns5P and PtdIns(3,5)P2 and implicated in insulin signaling, causes early embryonic death in mice. In contrast, mice with muscle-specific Pikfyve disruption have normal lifespan but exhibit early-age whole-body glucose intolerance and muscle insulin resistance, thus establishing the key role of muscle PIKfyve in glucose homeostasis. Fat and muscle tissues control postprandial glucose clearance through different mechanisms, raising questions as to whether adipose Pikfyve disruption will also trigger whole-body metabolic abnormalities, and if so, what the mechanism might be. To clarify these issues, here we have characterized two new mouse models with adipose tissue disruption of Pikfyve through Cre recombinase expression driven by adipose-specific aP2- or adiponectin (Aq) promoters. Whereas both mouse lines were ostensibly normal until adulthood, their glucose homeostasis and systemic insulin sensitivity were severely dysregulated. These abnormalities stemmed in part from accelerated fat-cell lipolysis and elevated serum FFA Intriguingly, aP2-Cre-PIKfyve(fl/fl) but not Aq-Cre-PIKfyve(fl/fl) females had severely impaired pregnancy-induced mammary gland differentiation and lactogenesis, consistent with aP2-Cre-mediated Pikfyve excision in nonadipogenic tissues underlying this defect. Intriguingly, whereas mammary glands from postpartum control and Aq-Cre-PIKfyve(fl/fl) mice or ex vivo mammary gland explants showed profound upregulation of PIKfyve protein levels subsequent to prolactin receptor activation, such increases were not apparent in aP2-Cre-PIKfyve(fl/fl) females. Collectively, our data identify for the first time that adipose tissue Pikfyve plays a key role in the mechanisms regulating glucose homeostasis and that the PIKfyve pathway is critical in mammary epithelial differentiation during pregnancy and lactogenesis downstream of prolactin receptor

  15. Raising Better Boys.

    ERIC Educational Resources Information Center

    Canada, Geoffrey

    2000-01-01

    The author of "Reaching Up For Manhood" discusses troubling social/environmental conditions confronting boys. Raising better boys requires caring adults, safer risk-taking situations, positive reinforcement, and role models. Parents should monitor boys' media exposure, provide moral education, broaden their cultural and natural-world…

  16. Raising Responsible Teenagers.

    ERIC Educational Resources Information Center

    Myers, Bob

    Some parents raise children without knowing anything about child development and are thus bewildered about how to deal with difficult behavior. This books addresses three important areas of parenting--child development, parenting style, and discipline--and tries to help parents gain the self-control needed to become competent adults. Chapters 1,…

  17. Raise Funds, Not Hackles.

    ERIC Educational Resources Information Center

    Zaidel, Lisa Brusman

    1996-01-01

    Ten strategies for fundraising include setting start/stop dates, dividing the workload, stating a purpose for the funds, establishing financial goals, choosing quality products to sell, remembering the sellers' ages, learning about the fund-raising companies, motivating the volunteers, considering a business sponsorship, and conducting a…

  18. Strategic Supply

    DTIC Science & Technology

    2006-01-01

    context of Supply Chain Management ( SCM ), it is quite apparent that Strategic Supply cannot be classified as a particular industry; but rather, as an...Management ( SCM ), it is quite apparent that Strategic Supply cannot be classified as a particular industry; but rather, as an enabler across all...advantage in the global marketplace. The Council of Supply Chain Management Professionals (CSCMP) has defined SCM as, “…encompassing the planning

  19. Strategic Materials

    DTIC Science & Technology

    2006-01-01

    Sears, US Navy Lt Col Mark Williams, US Air Force Dr. Sylvia Babus , Faculty Dr. Maureen Crandall, Faculty Mr. Bill Jones, Faculty Strategic...graying rapidly, populated predominantly with baby- boomers over 40 years of age. Aggravating this demographic shift, the number of workforce S&E...Transnational Environment. [Student Essay from Strategic Materials Industry Study 2005, available from Dr. Sylvia Babus , Industrial College of the Armed

  20. System's flips in climate-related energy (CRE) systems

    NASA Astrophysics Data System (ADS)

    Ramos, Maria-Helena; Creutin, Jean-Dominique; Engeland, Kolbjørn; François, Baptiste; Renard, Benjamin

    2014-05-01

    Several modern environmental questions invite to explore the complex relationships between natural phenomena and human behaviour at a range of space and time scales. This usually involves a number of cause-effect (causal) relationships, linking actions and events. In lay terms, 'effect' can be defined as 'what happened' and 'cause', 'why something happened.' In a changing world or merely moving from one scale to another, shifts in perspective are expected, bringing some phenomena into the foreground and putting others to the background. Systems can thus flip from one set of causal structures to another in response to environmental perturbations and human innovations or behaviors, for instance, as space-time signatures are modified. The identification of these flips helps in better understanding and predicting how societies and stakeholders react to a shift in perspective. In this study, our motivation is to investigate possible consequences of the shift to a low carbon economy in terms of socio-technico systems' flips. The focus is on the regional production of Climate-Related Energy (CRE) (hydro-, wind- and solar-power). We search for information on historic shifts that may help defining the forcing conditions of abrupt changes and extreme situations. We identify and present a series of examples in which we try to distinguish the various tipping points, thresholds, breakpoints and regime shifts that are characteristic of complex systems in the CRE production domain. We expect that with these examples our comprehension of the question will be enriched, providing us the elements needed to better validate modeling attempts, to predict and manage flips of complex CRE production systems. The work presented is part of the FP7 project COMPLEX (Knowledge based climate mitigation systems for a low carbon economy; http://www.complex.ac.uk/).

  1. Risk Factors for Carbapenemase-Producing Carbapenem-Resistant Enterobacteriaceae (CP-CRE) Acquisition Among Contacts of Newly Diagnosed CP-CRE Patients.

    PubMed

    Schwartz-Neiderman, Anat; Braun, Tali; Fallach, Noga; Schwartz, David; Carmeli, Yehuda; Schechner, Vered

    2016-10-01

    OBJECTIVE Carbapenemase-producing carbapenem-resistant Enterobacteriaceae (CP-CRE) are extremely drug-resistant pathogens. Screening of contacts of newly identified CP-CRE patients is an important step to limit further transmission. We aimed to determine the risk factors for CP-CRE acquisition among patients exposed to a CP-CRE index patient. METHODS A matched case-control study was performed in a tertiary care hospital in Israel. The study population was comprised of patients who underwent rectal screening for CP-CRE following close contact with a newly identified CP-CRE index patient. Cases were defined as positive tests for CP-CRE. For each case patient, 2 matched controls were randomly selected from the pool of contacts who tested negative for CP-CRE following exposure to the same index case. Bivariate and multivariate analyses were conducted using conditional logistic regression. RESULTS In total, 53 positive contacts were identified in 40 unique investigations (896 tests performed on 735 contacts) between October 6, 2008, and June 7, 2012. bla KPC was the only carbapenemase identified. In multivariate analysis, risk factors for CP-CRE acquisition among contacts were (1) contact with an index patient for ≥3 days (odds ratio [OR], 9.8; 95% confidence interval [CI], 2.0-48.9), (2) mechanical ventilation (OR, 4.1; 95% CI, 1.4-11.9), and (3) carriage or infection with another multidrug-resistant organism (MDRO; OR, 2.6; 95% CI, 1.0-7.1). Among patients who received antibiotics, cephalosporins were associated with a lower risk of acquisition. CONCLUSIONS Patient characteristics (ventilation and carriage of another MDRO) as well as duration of contact are risk factors for CP-CRE acquisition among contacts. The role of cephalosporins requires further study. Infect Control Hosp Epidemiol 2016;1-7.

  2. Generation of Cre Transgenic Mice with Postnatal RPE-Specific Ocular Expression

    PubMed Central

    Iacovelli, Jared; Zhao, Chen; Wolkow, Natalie; Veldman, Peter; Gollomp, Kandace; Ojha, Pallavi; Lukinova, Nina; King, Ayala; Feiner, Leonard; Esumi, Noriko; Zack, Donald J.; Pierce, Eric A.; Vollrath, Douglas

    2011-01-01

    Purpose. To generate and characterize a constitutively active, RPE-specific, cre-expressing transgenic mouse line. This line can be used to create RPE-specific knockouts by crossing with mice harboring loxP-flanked (floxed) genes. Methods. A transgene construct was assembled with the BEST1 promoter driving cre expression. Transgenic mice were generated on a C57BL/6 background. Cre expression was assessed by immunofluorescence and Western blot analysis. Cre enzymatic activity was tested by crossing to three lines with floxed DNA regions and detecting deletion of the intervening sequences or through histochemical detection of lacZ activity. Potential cre-mediated toxicity was assessed by retinal histology up to 24 months of age and by electroretinography. Results. The BEST1-cre line with expression in the highest percentage of RPE cells displayed a patchy mosaic expression pattern, with 50% to 90% of RPE cells expressing cre. In mice outcrossed to a mixed B6/129 background, expression was consistently found in 90% of RPE cells. Within the eye, only the RPE cells were immunoreactive with an anti-cre antibody. Maximum cre expression quantified by Western blot analysis occurred at P28. Crosses with three lines containing floxed sequences revealed RPE-specific cre activity in the eye and extraocular expression limited to the testes. Histology and electroretinography showed no cre-mediated RPE toxicity. Conclusions. This BEST1-cre transgenic line enables generation of RPE-specific knockout mice. The mosaic expression pattern provides an internal control; the non–cre-expressing RPE cells continue to express the floxed genes. These mice should facilitate study of the multifunctional RPE and the generation of mouse models of human retinal disease. PMID:21212186

  3. Cre-Mediated Recombination in Rhombic Lip Derivatives

    PubMed Central

    Fünfschilling, Ursula; Reichardt, Louis F.

    2009-01-01

    Summary To study the development of the cerebellum, we generated a transgenicmouse line Tg(mα6-cre)B1LFR that expresses CRE recombinase under the GABAA receptor α6 subunit promoter. In this line, recombination of an R26R reporter allele occurred postnatally in granule cells of the cerebellum and dorsal cochlear nucleus, as well as in a subset of precerebellar nuclei in the brainstem. All neurons in which recombination occurred originated during embryogenesis from the rhombic lip. This might be explained by a very early specification event at the rhombic lip that primes cells derived from this structure to express the transgene during neuronal maturation. As no recombination occurred in the inferior olive, it may be derived from a distinct subset of precursors at the rhombic lip. No recombination occurred in any of the interneurons in the cerebellum (stellate cells, basket cells, and Golgi cells), consistent with the hypothesis that they are not derived from the same embryonic tissue as the granule cells. genesis 33:160–169, 2002. PMID:12203913

  4. The Legacy of Nat Sternberg: The Genesis of Cre-lox Technology.

    PubMed

    Yarmolinsky, Michael; Hoess, Ronald

    2015-11-01

    Cre-lox of bacteriophage P1 has become one of the most widely used tools for genetic engineering in eukaryotes. The origins of this tool date to more than 30 years ago when Nat L. Sternberg discovered the recombinase, Cre, and its specific locus of crossover, lox, while studying the maintenance of bacteriophage P1 as a stable plasmid. Recombinations mediated by Cre assist in cyclization of the DNA of infecting phage and in resolution of prophage multimers created by generalized recombination. Early in vitro work demonstrated that, although it shares similarities with the well-characterized bacteriophage λ integration, Cre-lox is in many ways far simpler in its requirements for carrying out recombination. These features would prove critical for its development as a powerful and versatile tool in genetic engineering. We review the history of the discovery and characterization of Cre-lox and touch upon the present direction of Cre-lox research.

  5. Cre recombinase induces DNA damage and tetraploidy in the absence of loxP sites.

    PubMed

    Janbandhu, Vaibhao C; Moik, Daniel; Fässler, Reinhard

    2014-01-01

    The spatiotemporal manipulations of gene expression by the Cre recombinase (Cre) of bacteriophage P1 has become an essential asset to understanding mammalian genetics. Accumulating evidence suggests that Cre activity can, in addition to excising targeted loxP sites, induce cytotoxic effects, including abnormal cell cycle progression, genomic instability, and apoptosis, which can accelerate cancer progression. It is speculated that these defects are caused by Cre-induced DNA damage at off-target sites. Here we report the formation of tetraploid keratinocytes in the epidermis of keratin 5 and/or keratin 14 promoter-driven Cre (KRT5- and KRT14-Cre) expressing mouse skin. Biochemical analyses and flow cytometry demonstrated that Cre expression also induces DNA damage, genomic instability, and tetraploidy in HCT116 cells, and live-cell imaging revealed an extension of the G 2 cell cycle phase followed by defective or skipping of mitosis as cause for the tetraploidy. Since tetraploidy eventually leads to aneuploidy, a hallmark of cancer, our findings highlight the importance of distinguishing non-specific cytopathic effects from specific Cre/loxP-driven genetic manipulations when using Cre-mediated gene deletions.

  6. Tamoxifen-Containing Eye Drops Successfully Trigger Cre-Mediated Recombination in the Entire Eye.

    PubMed

    Schlecht, Anja; Leimbeck, Sarah V; Tamm, Ernst R; Braunger, Barbara M

    2016-01-01

    Embryonic lethality in mice with targeted gene deletion is a major issue that can be circumvented by using Cre-loxP-based animal models. Various inducible Cre systems are available, e.g. such that are activated following tamoxifen treatment, and allow deletion of a specific target gene at any desired time point during the life span of the animal. In this study, we describe the efficiency of topical tamoxifen administration by eye drops using a Cre- reporter mouse strain (R26R). We report that tamoxifen-responsive CAGGCre-ER (TM) mice show a robust Cre- mediated recombination throughout the entire eye.

  7. Cre/ lox site-specific recombination controls the excision of a transgene from the rice genome.

    PubMed

    Hoa, T. T. C.; Bong, B. B.; Huq, E.; Hodges, T. K.

    2002-03-01

    The Cre/ lox site-specific recombination controls the excision of a target DNA segment by recombination between two loxsites flanking it, mediated by the Cre recombinase. We have studied the functional expression of the Cre/ lox system to excise a transgene from the rice genome. We developed transgenic plants carrying the target gene, hygromycin phosphotransferase ( hpt) flanked by two lox sites and transgenic plants harboring the Cre gene. Each lox plant was crossed with each Cre plant reciprocally. In the Cre /lox hybrid plants, the Cre recombinase mediates recombination between two lox sites, resulting in excision of the hpt gene. The recombination event could be detected because it places the CaMV 35S promoter of the hpt gene adjacent to a promoterless gusA gene; as a result the gusA gene is activated and its expression could be visualized. In 73 Cre /lox hybrid plants from various crosses of T0 transgenic plants, 19 expressed GUS, and in 132 Cre /lox hybrid plants from crosses of T2 transgenic plants, 77 showed GUS expression. Molecular data proved the excision event occurred in all the GUS(+) plants. Recombination occurred with high efficiency at the early germinal stage, or randomly during somatic development stages.

  8. Chromosomal localisation of the CD4cre transgene in B6·Cg-Tg(Cd4-cre)1Cwi mice.

    PubMed

    Westendorf, Kerstin; Durek, Pawel; Ayew, Samia; Mashreghi, Mir-Farzin; Radbruch, Andreas

    2016-09-01

    The B6·Cg-Tg(Cd4-cre)1Cwi line expresses CRE recombinase under the control of the promoter and regulatory elements of the Cd4 gene. Here we show that CRE recombinase expression reduces the number and frequencies of CD4 positive subsets in a dose-dependent manner and localize the integration site of the transgenic construct to position 60335693-60341285 (qD) of chromosome 3. The insert contains at least 15 complete sequential copies of the transgenic construct. Based on this information we describe a novel PCR assay for genetic typing of transgenic mice.

  9. Strategic Staffing

    ERIC Educational Resources Information Center

    Clark, Ann B.

    2012-01-01

    Business and industry leaders do not flinch at the idea of placing top talent in struggling departments and divisions. This is not always the case in public education. The Charlotte-Mecklenburg Schools made a bold statement to its community in its strategic plan by identifying two key reform levers--(1) an effective principal leading each school;…

  10. Strategic HRD.

    ERIC Educational Resources Information Center

    1997

    This document contains four papers from a symposium on strategic human resource development (HRD). "Examination of the Use of Scenarios as Learning and Decision-Making Tools for Human Resource Development" (Roger F. Miller, Susan A. Lynham, Joanne Provo, Jeanne M. St. Claire) explores the potential value of scenarios as learning and…

  11. Fund-raising fiasco.

    PubMed

    1997-05-16

    The Pennsylvania Attorney General's office was prepared to file a civil suit against three local AIDS agencies and a fund-raiser organization that they hired to solicit donations. The suit would have charged the fund-raiser, Pallotta and Associates of Los Angeles, with misrepresenting to donors that 60 percent of the Philadelphia-DC AIDS Ride would go to beneficiaries. The organizations agreed to pay $134,000 to settle claims that they violated the law. Organizers had hoped that 1,600 cyclists would participate in the AIDS Ride event; only 790 actually took part. The riders raised $1.6 million, yielding only a 16 percent profit for the AIDS groups. The proceeds of an out-of-court settlement will be disbursed to charities.

  12. Retina-specific gene excision by targeted expression of Cre recombinase.

    PubMed

    Ding, Yuzhi; Li, Jianmin; Liu, Qinghuai

    2013-11-29

    The use of Cre recombinase for conditional targeting permits the controlled removal or activation of genes in specific tissues and at specific times of development. The Rho-Cre mice provide an improved tool for studying gene ablation in rod photoreceptor cells. To establish a robust expression of Rho-Cre transgenic mice that would be useful for the study of various protein functions in photoreceptor cells, a total 11,987 kb fragment (pNCHS4 Rho-NLS-cre) containing human rhodopsin promoter was cloned. The Rho-Cre plasmid was digested with EcoR1 and I Ceu-1, and the 9.316 kb fragment containing the hRho promoter and Cre recombinase gel was purified. To generate transgenic mice, the purified DNA fragment was injected into fertilized oocytes according to standard protocols. ROSA26R reported the steady expression of Rho-Cre especially in photoreceptor cells, allowing further excising proteins in rod photoreceptors across the retina. This Rho-Cre transgenic line should thus prove useful as a general deletor line for genetic analysis of diverse aspects of retinopathy.

  13. Effects of CreERT2, 4-OH Tamoxifen, and Gender on CFU-F Assays

    PubMed Central

    McHaffie, Sophie L.; Hastie, Nicholas D.; Chau, You-Ying

    2016-01-01

    Gene function in stem cell maintenance is often tested by inducing deletion via the Cre-loxP system. However, controls for Cre and other variables are frequently not included. Here we show that when cultured in the presence of 4-OH tamoxifen, bone and marrow cells containing the CreERT2 construct have a reduced colony forming ability. Inactive CreERT2 recombinase, however, has the opposite effect. Young female marrow cells containing the inactive CreERT2 construct grew more colonies than cells lacking the construct altogether. Young female control marrow cells (i.e., negative for CreERT2) also produced significantly greater colony numbers when cultured with 4-OH tamoxifen, compared with the ethanol vehicle control. In conclusion, we report that the use of the Cre-loxP system is inadvisable in combination with CFU-F assays, and that appropriate controls should be in place to extend the future use of Cre-loxP in alternate assays. PMID:26828722

  14. Effects of CreERT2, 4-OH Tamoxifen, and Gender on CFU-F Assays.

    PubMed

    McHaffie, Sophie L; Hastie, Nicholas D; Chau, You-Ying

    2016-01-01

    Gene function in stem cell maintenance is often tested by inducing deletion via the Cre-loxP system. However, controls for Cre and other variables are frequently not included. Here we show that when cultured in the presence of 4-OH tamoxifen, bone and marrow cells containing the CreERT2 construct have a reduced colony forming ability. Inactive CreERT2 recombinase, however, has the opposite effect. Young female marrow cells containing the inactive CreERT2 construct grew more colonies than cells lacking the construct altogether. Young female control marrow cells (i.e., negative for CreERT2) also produced significantly greater colony numbers when cultured with 4-OH tamoxifen, compared with the ethanol vehicle control. In conclusion, we report that the use of the Cre-loxP system is inadvisable in combination with CFU-F assays, and that appropriate controls should be in place to extend the future use of Cre-loxP in alternate assays.

  15. Efficient BLG-Cre mediated gene deletion in the mammary gland.

    PubMed

    Selbert, S; Bentley, D J; Melton, D W; Rannie, D; Lourenço, P; Watson, C J; Clarke, A R

    1998-09-01

    Using the phage P1-derived Cre/loxP recombination system, we have developed a strategy for efficient mammary tissue specific inactivation of floxed genes. Transgenic mice were generated which express Cre DNA-recombinase under the control of the mammary gland specific promoter of the ovine beta-lactoglobulin (BLG) gene. To test the specificity of Cre mediated recombination, we crossed these mice to animals harbouring a floxed DNA ligase I allele. We show that the BLG-Cre construct specifies mammary specific gene deletion, and furthermore that it is temporally regulated, predominantly occurring during lactation. We fully characterised the extent of gene deletion in one line (line 74). In this strain the virgin gland is characterised by low levels (7%) of Cre mediated deletion, whereas 70-80% of cells within the lactating mammary gland have undergone recombination. Immunohistochemistry and indirect in situ PCR were used respectively to demonstrate that both Cre protein and Cre activity were evenly distributed throughout the population of secretory epithelial cells. The level of background recombination in non-mammary tissues was found to be < or = 1.1%, irrespective of mammary gland developmental status. Crossing the transgenic BLG-Cre strain described here to mice harbouring other floxed alleles will facilitate the functional analysis of those genes during differentiation and development of the mammary gland.

  16. Dual Fluorescent Reporter Pig for Cre Recombination: Transgene Placement at the ROSA26 Locus

    PubMed Central

    Li, Shun; Flisikowska, Tatiana; Kurome, Mayuko; Zakhartchenko, Valeri; Kessler, Barbara; Saur, Dieter; Kind, Alexander; Wolf, Eckhard; Flisikowski, Krzysztof; Schnieke, Angelika

    2014-01-01

    We are extending the Cre/loxP site-specific recombination system to pigs, focussing on conditional and tissue-specific expression of oncogenic mutations to model human cancers. Identifying the location, pattern and extent of Cre recombination in vivo is an important aspect of this technology. Here we report pigs with a dual fluorochrome cassette under the control of the strong CAG promoter that switches expression after Cre-recombination, from membrane-targeted tandem dimer Tomato to membrane-targeted green fluorescent protein. The reporter cassette was placed at the porcine ROSA26 locus by conventional gene targeting using primary mesenchymal stem cells, and animals generated by nuclear transfer. Gene targeting efficiency was high, and analysis of foetal organs and primary cells indicated that the reporter is highly expressed and functional. Cre reporter pigs will provide a multipurpose indicator of Cre recombinase activity, an important new tool for the rapidly expanding field of porcine genetic modification. PMID:25025770

  17. Raising the continental crust

    NASA Astrophysics Data System (ADS)

    Campbell, Ian H.; Davies, D. Rhodri

    2017-02-01

    that the removal of 40 km of the amphibole-garnet-pyroxenite root would have raised the average level of the continental crust by ∼3 km. The emergence of the continental crust was an essential precursor to the rise of oxygen, which started some 200 Myr later.

  18. Spinal Hb9::Cre-derived excitatory interneurons contribute to rhythm generation in the mouse

    PubMed Central

    Caldeira, Vanessa; Dougherty, Kimberly J.; Borgius, Lotta; Kiehn, Ole

    2017-01-01

    Rhythm generating neurons are thought to be ipsilaterally-projecting excitatory neurons in the thoracolumbar mammalian spinal cord. Recently, a subset of Shox2 interneurons (Shox2 non-V2a INs) was found to fulfill these criteria and make up a fraction of the rhythm-generating population. Here we use Hb9::Cre mice to genetically manipulate Hb9::Cre-derived excitatory interneurons (INs) in order to determine the role of these INs in rhythm generation. We demonstrate that this line captures a consistent population of spinal INs which is mixed with respect to neurotransmitter phenotype and progenitor domain, but does not overlap with the Shox2 non-V2a population. We also show that Hb9::Cre-derived INs include the comparatively small medial population of INs which continues to express Hb9 postnatally. When excitatory neurotransmission is selectively blocked by deleting Vglut2 from Hb9::Cre-derived INs, there is no difference in left-right and/or flexor-extensor phasing between these cords and controls, suggesting that excitatory Hb9::Cre-derived INs do not affect pattern generation. In contrast, the frequencies of locomotor activity are significantly lower in cords from Hb9::Cre-Vglut2Δ/Δ mice than in cords from controls. Collectively, our findings indicate that excitatory Hb9::Cre-derived INs constitute a distinct population of neurons that participates in the rhythm generating kernel for spinal locomotion. PMID:28128321

  19. Cardiac-specific activation of Cre expression at late fetal development

    SciTech Connect

    Opherk, Jan P.; Yampolsky, Peter; Hardt, Stefan E.; Schoels, Wolfgang; Katus, Hugo A.; Koenen, Michael . E-mail: koenen@mpimf-heidelberg.mpg.de; Zehelein, Joerg

    2007-07-27

    In a first step towards dissecting molecular mechanisms that contribute to the development of cardiac diseases, we have generated transgenic mice that express a Cre-GFP fusion protein under the transcriptional control of a 4.3 kb murine cardiac Troponin I gene (cTnI) promoter. Cre-GFP expression, similar in three transgenic lines, is described in one line. In mouse embryos, transgenic for the Cre-GFP and ROSA lacZ reporter allele, first Cre-mediated recombination appeared at 16.5 dpc selectively at the heart. Like the endogenous cTnI gene, transgenic Cre expression showed a slow rise through fetal development that increased neonatally. Bitransgenic hearts, stained at 30 days of age, showed intense signals in ventricular and atrial myocytes while no recombination occurred in other tissues. The delayed onset of Cre activity in cTnI-Cre mice could provide a useful genetic tool to evaluate the function of loxP targeted cardiac genes without interference of recombination during early heart development.

  20. Advanced zebrafish transgenesis with Tol2 and application for Cre/lox recombination experiments.

    PubMed

    Mosimann, Christian; Zon, Leonard I

    2011-01-01

    Spatio-temporal transgene regulation by transgenic DNA recombinases is a central tool for genetic research in multicellular organisms, with excellent applications for misexpression and lineage tracing experiments. Cre recombinase-controlled lox site recombination is a cornerstone of contemporary mouse genetics, and Cre/lox techniques therefore attract increasing interest in the zebrafish field. Tol2-mediated zebrafish transgenesis now provides a stable platform for lox cassette transgenes, while the ease of drug treatments in zebrafish makes the model an ideal candidate for Tamoxifen/4-hydroxytamoxifen-inducible CreER(T2) experiments. In this chapter, we will first introduce the basics of Cre/lox methodology, CreER(T2) regulation by Tamoxifen/4-hydroxytamoxifen, as well as the benefits of Tol2 transgenesis for Cre/lox experiments. We will then in detail outline practical experimental steps for Tol2 transgenesis toward the creation of single-insertion transgenes. Lastly, we will introduce protocols for 4-hydroxytamoxifen-mediated CreER(T2) induction to perform spatio-temporal lox transgene regulation experiments in zebrafish embryos.

  1. Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach.

    PubMed

    Jungke, Peggy; Hammer, Juliane; Hans, Stefan; Brand, Michael

    2015-01-01

    Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre-dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome.

  2. Strategic Petroleum Reserves

    SciTech Connect

    Not Available

    1989-01-01

    GAO examined approximately 40 alternative, nontraditional methods of acquiring oil for the Strategic Petroleum Reserve. The alternatives identified were compared to the current method of acquiring and financing SPR oil through congressional appropriations that are reported in the budget. When compared to the conventional method of financing oil, most of the proposals have certain benefits or advantages, but all of them have economic or other disadvantages. For example, some proposals would reduce the budget deficit by increasing government revenues but would raise prices to the consumer. Other proposals would reduce short-term expenditures (asset sales, leasing oil or indexed bonds). However, the proposals might increase long-term expenditures by more than the initial reduction in outlays.

  3. Strategic petroleum reserves

    SciTech Connect

    Not Available

    1989-04-01

    This report has examined approximately 40 alternative, nontraditional methods of acquiring oil for the Strategic Petroleum Reserve. The alternatives identified are compared to the current method of acquiring and financing SPR oil through congressional appropriations that are reported in the budget. When compared to the conventional method of financing oil, most of the proposals have certain benefits or advantages, but all of them have economic or other disadvantages. For example, some proposals would reduce the budget deficit by increasing government revenues but would raise prices to the consumer. Other proposals would reduce short-term expenditures (asset sales, leasing oil or indexed bonds). However, the proposals might increase long-term expenditures by more than the initial reduction in outlays.

  4. On Strategic Management: A Conversation with George Wilkinson.

    ERIC Educational Resources Information Center

    Brandt, Ron

    1991-01-01

    George Wilkinson, who has managed planning for United Way of America since 1980, observes that local planners using strategic planning raise more money and solve more community problems than those without such a process. Marketing is identical to strategic planning; both involve looking at an organization, doing and analysis, examining the future,…

  5. aP2-Cre-Mediated Inactivation of Estrogen Receptor Alpha Causes Hydrometra

    PubMed Central

    Antonson, Per; Matic, Marko; Portwood, Neil; Kuiper, Raoul V.; Bryzgalova, Galyna; Gao, Hui; Windahl, Sara H.; Humire, Patricia; Ohlsson, Claes; Berggren, Per-Olof; Gustafsson, Jan-Åke; Dahlman-Wright, Karin

    2014-01-01

    In this study we describe the reproductive phenotypes of a novel mouse model in which Cre-mediated deletion of ERα is regulated by the aP2 (fatty acid binding protein 4) promoter. ERα-floxed mice were crossed with transgenic mice expressing Cre-recombinase under the control of the aP2 promoter to generate aP2-Cre/ERαflox/flox mice. As expected, ERα mRNA levels were reduced in adipose tissue, but in addition we also detected an 80% reduction of ERα levels in the hypothalamus of aP2-Cre/ERαflox/flox mice. Phenotypic analysis revealed that aP2-Cre/ERαflox/flox female mice were infertile. In line with this, aP2-Cre/ERαflox/flox female mice did not cycle and presented 3.8-fold elevated estrogen levels. That elevated estrogen levels were associated with increased estrogen signaling was evidenced by increased mRNA levels of the estrogen-regulated genes lactoferrin and aquaporin 5 in the uterus. Furthermore, aP2-Cre/ERαflox/flox female mice showed an accumulation of intra-uterine fluid, hydrometra, without overt indications for causative anatomical anomalies. However, the vagina and cervix displayed advanced keratosis with abnormal quantities of accumulating squamous epithelial cells suggesting functional obstruction by keratin plugs. Importantly, treatment of aP2-Cre/ERαflox/flox mice with the aromatase inhibitor Letrozole caused regression of the hydrometra phenotype linking increased estrogen levels to the observed phenotype. We propose that in aP2-Cre/ERαflox/flox mice, increased serum estrogen levels cause over-stimulation in the uterus and genital tracts resulting in hydrometra and vaginal obstruction. PMID:24416430

  6. Thinking strategically.

    PubMed

    Goree, Michael

    2002-01-01

    Over the course of the past 20 years, human resources has tried a variety of strategic initiatives to add value to the working environment, from the alphabets of TQM, CQI, EVA, ROI, ISO, QS, Theory X, Y, Z, Generation X and Y to re-engineering, balanced scorecard, lean, hoshin, six sigma, to Margaret Wheatley's "The Simpler Way" and finally to cheese and fish. The problem is that none of these is a strategy. They are all tactics to accomplish or achieve a strategy.

  7. Expression of Cre recombinase during transient phage infection permits efficient marker removal in Streptomyces

    PubMed Central

    Khodakaramian, Gholam; Lissenden, Sarah; Gust, Bertolt; Moir, Laura; Hoskisson, Paul A.; Chater, Keith F.; Smith, Margaret C. M.

    2006-01-01

    We report a system for the efficient removal of a marker flanked by two loxP sites in Streptomyces coelicolor, using a derivative of the temperate phage φC31 that expresses Cre recombinase during a transient infection. As the test case for this recombinant phage (called Cre-phage), we present the construction of an in-frame deletion of a gene, pglW, required for phage growth limitation or Pgl in S.coelicolor. Cre-phage was also used for marker deletion in other strains of S.coelicolor. PMID:16473843

  8. Bigenic Cre/loxP, puΔtk conditional genetic ablation

    PubMed Central

    Chen, You-Tzung; Levasseur, Regis; Vaishnav, Sukeshi; Karsenty, Gerard; Bradley, Allan

    2004-01-01

    Genetic ablation experiments are used to resolve problems regarding cell lineages and the in vivo function of certain groups of cells. We describe a two-component conditional ablation technology using a mouse carrying an X-linked puΔtk transgene, which is only activated in cells expressing Cre. Ablation of the Cre-expressing cells can be temporally regulated by the time of ganciclovir (GCV) administration. This strategy was demonstrated using a Col2Cre transgenic line. Differentiating chondrocytes in bigenic animals could be ablated at different developmental stages resulting in disorganized growth plates and dwarfism. Macrocephaly, macroglossia and umbilical hernia were also observed in ablated 18.5 dpc embryos. Crosses between the puΔtk selector transgenic line and existing cre lines will facilitate numerous temporally regulated tissue-specific ablation experiments. PMID:15561996

  9. CDC Vital Signs: Making Health Care Safer -- Stop Infections from Lethal CRE Germs Now

    MedlinePlus

    ... 62 MB] Read the MMWR Science Clips Making Health Care Safer Stop Infections from Lethal CRE Germs Now ... to otherwise healthy people outside of medical facilities. Health Care Providers can Know if patients in your facility ...

  10. Screening for novel human genes associated with CRE pathway activation with cell microarray.

    PubMed

    Tian, Linjie; Wang, Pingzhang; Guo, Jinhai; Wang, Xinyu; Deng, Weiwei; Zhang, Chenying; Fu, Dongxu; Gao, Xia; Shi, Taiping; Ma, Dalong

    2007-07-01

    In this study, cell microarray technology is used to identify novel human genes associated with CRE pathway activation. By reverse transfection, expression plasmids containing full-length cDNAs were cotransfected with the reporter plasmid pCRE-d2EGFP to monitor the activation of the CRE pathway via enhanced green fluorescence protein (EGFP) expression. Of the 575 predominantly novel genes screened, 22 exhibited relatively higher EGFP fluorescence compared with a negative control. After a functional validation with a dual luciferase reporter system that included both cis- and trans-luciferase assays, 4 of the 22 genes (RNF41, C8orf32, C6orf208, and MEIS3P1) were confirmed as CRE-pathway activators. Western blot analysis revealed that RNF41 can promote CREB phosphorylation. These results demonstrate the successful combination of cell microarray technology with this reporting system and the potential of this tool to characterize functions of novel genes in a highly parallel format.

  11. Inducible Cre transgenic mouse strain for skeletal muscle-specific gene targeting

    PubMed Central

    2012-01-01

    Background The use of the Cre/loxP system for gene targeting has been proven to be a powerful tool for understanding gene function. The purpose of this study was to create and characterize an inducible, skeletal muscle-specific Cre transgenic mouse strain. Methods To achieve skeletal muscle-specific expression, the human α-skeletal actin promoter was used to drive expression of a chimeric Cre recombinase containing two mutated estrogen receptor ligand-binding domains. Results Western blot analysis, PCR and β-galactosidase staining confirmed that Cre-mediated recombination was restricted to limb and craniofacial skeletal muscles only after tamoxifen administration. Conclusions A transgenic mouse was created that allows inducible, gene targeting of floxed genes in adult skeletal muscle of different developmental origins. This new mouse will be of great utility to the skeletal muscle community. PMID:22564549

  12. Strategic Planning: What's so Strategic about It?

    ERIC Educational Resources Information Center

    Strong, Bart

    2005-01-01

    The words "strategic" and "planning" used together can lead to confusion unless one spent the early years of his career in never-ending, team-oriented, corporate training sessions. Doesn't "strategic" have something to do with extremely accurate bombing or a defensive missile system or Star Wars or something? Don't "strategic" and "planning" both…

  13. Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach

    PubMed Central

    Jungke, Peggy; Hammer, Juliane; Hans, Stefan; Brand, Michael

    2015-01-01

    Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre–dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome. PMID:26083735

  14. Inducible targeting of CNS astrocytes in Aldh1l1-CreERT2 BAC transgenic mice

    PubMed Central

    Winchenbach, Jan; Düking, Tim; Berghoff, Stefan A.; Stumpf, Sina K.; Hülsmann, Swen; Nave, Klaus-Armin; Saher, Gesine

    2016-01-01

    Background: Studying astrocytes in higher brain functions has been hampered by the lack of genetic tools for the efficient expression of inducible Cre recombinase throughout the CNS, including the neocortex. Methods: Therefore, we generated BAC transgenic mice, in which CreERT2 is expressed under control of the Aldh1l1 regulatory region. Results: When crossbred to Cre reporter mice, adult Aldh1l1-CreERT2 mice show efficient gene targeting in astrocytes. No such Cre-mediated recombination was detectable in CNS neurons, oligodendrocytes, and microglia. As expected, Aldh1l1-CreERT2 expression was evident in several peripheral organs, including liver and kidney. Conclusions: Taken together, Aldh1l1-CreERT2 mice are a useful tool for studying astrocytes in neurovascular coupling, brain metabolism, synaptic plasticity and other aspects of neuron-glia interactions. PMID:28149504

  15. Generation of a tamoxifen inducible Tnnt2MerCreMer knock-in mouse model for cardiac studies.

    PubMed

    Yan, Jianyun; Sultana, Nishat; Zhang, Lu; Park, David S; Shekhar, Akshay; Hu, Jun; Bu, Lei; Cai, Chen-Leng

    2015-06-01

    Tnnt2, encoding thin-filament sarcomeric protein cardiac troponin T, plays critical roles in heart development and function in mammals. To develop an inducible genetic deletion strategy in myocardial cells, we generated a new Tnnt2:MerCreMer (Tnnt2(MerCreMer/+)) knock-in mouse. Rosa26 reporter lines were used to examine the specificity and efficiency of the inducible Cre recombinase. We found that Cre was specifically and robustly expressed in the cardiomyocytes at embryonic and adult stages following tamoxifen induction. The knock-in allele on Tnnt2 locus does not impact cardiac function. These results suggest that this new Tnnt2(MerCreMer/+) mouse could be applied towards the temporal genetic deletion of genes of interests in cardiomyocytes with Cre-LoxP technology. The Tnnt2(MerCreMer/+) mouse model also provides a useful tool to trace myocardial lineage during development and repair after cardiac injury.

  16. Cre Recombinase: You Can't Live with It, and You Can't Live Without It.

    PubMed

    Le, Yun-Zheng; Zhu, Meili; Anderson, Robert E

    2016-01-01

    The development of conditional gene targeting has greatly advanced our knowledge of human retinal diseases, but issues have arisen related to the use of some Cre-expressing mouse lines. In this article, we discuss potential problems associated with transgenic Cre expression-induced degeneration and alteration of rod photoreceptors and retinal pigment epithelium (RPE). Our strategy for circumventing RPE degeneration by induced transient Cre expression uses a single intravitreal doxycycline injection in a tetracycline-inducible RPE-specific Cre mouse line, which results in productive Cre-mediated recombination efficiently in the RPE. As constitutive expression of Cre in the RPE alters RPE biology, this inducible Cre/lox system provides an opportunity for conditional gene targeting in the RPE, a tissue that is closely related to photoreceptor degeneration, age-related macular degeneration, and diabetic retinopathy.

  17. Genetic inducible fate mapping in adult mice using tamoxifen-dependent Cre recombinases.

    PubMed

    Feil, Susanne; Krauss, Jana; Thunemann, Martin; Feil, Robert

    2014-01-01

    The Cre/lox site-specific recombination system allows the control of gene activity in space and time in almost any tissue of the mouse. A major technical advance was the development of tamoxifen-dependent Cre recombinases, such as CreER(T2), that can be activated by administration of tamoxifen to the animal. This powerful tool greatly facilitates the study of gene functions and the generation of more realistic animal models of sporadic human diseases. Another important application of tamoxifen-dependent Cre recombinases is genetic inducible fate mapping (GIFM). In GIFM studies, the inducible Cre/lox system is used to genetically label a defined cell population at a selected time by irreversible activation of the expression of a Cre-responsive reporter transgene. Then, marked cells are detected at later time points to determine how the originally labeled progenitors contribute to specific structures and cell types during pre- and postnatal development. GIFM was initially applied during mouse embryogenesis, but is now increasingly used for cell lineage tracing in adult mice under physiological and pathophysiological conditions. Here we describe the design of GIFM experiments in adult mice as exemplified by CreER(T2)-assisted tracing of vascular smooth muscle cells during the development of atherosclerotic lesions. First, we give an overview of reporter transgenes available for genetic cell marking that are expressed from the Rosa26 locus, such as β-galactosidase and fluorescent proteins. Then we present detailed protocols for the generation of experimental mice for GIFM studies, the induction of cell labeling by tamoxifen treatment, and the detection of marked cells in fixed and live tissues. Each section also provides a discussion of limitations and common pitfalls of GIFM experiments. Most of the protocols can be easily adapted to other developmental stages, cell types, Cre recombinases, and reporter transgenes and, thus, can be used as general guidelines for GIFM

  18. String Vacuous Overt Verb Raising.

    ERIC Educational Resources Information Center

    Koisumi, Masatoshi

    2000-01-01

    Presents evidence for overt verb raising in Japanese, thereby defending the validity of the head parameter with its two values (head initial and head final). Explores consequences of the overt verb raising analysis of Japanese to various aspects of syntactic theory. (Author/VWL)

  19. ELT and Consciousness-Raising

    ERIC Educational Resources Information Center

    Al-Jardani, Khalid Salim Saif

    2012-01-01

    The paper highlights the concept of consciousness-raising. It relates it to different aspects of ELT such as explicit teaching, language awareness, language acquisition and practice. How these terms are related to the concept of consciousness-raising within the English Language teaching. Its main aim is to help learners to notice for themselves…

  20. Regulation of Cre recombinase by ligand-induced complementation of inactive fragments

    PubMed Central

    Jullien, Nicolas; Sampieri, François; Enjalbert, Alain; Herman, Jean-Paul

    2003-01-01

    Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. To overcome this, we have developed DiCre, a regulatable fragment complementation system for Cre. The enzyme was split into two moieties that were fused to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12–rapamycin-associated protein), respectively. These can be efficiently heterodimerized by rapamycin. Several variants, based on splitting Cre at different sites and using different linker peptides, were tested in an indicator cell line. The fusion proteins, taken separately, had no recombinase activity. Stable transformants, co-expressing complementing fragments based on splitting Cre between Asn59 and Asn60, displayed low background activity affecting 0.05–0.4% of the cells. Rapamycin induced a rapid recombination, reaching 100% by 48–72 h, with an EC50 of 0.02 nM. Thus, ligand-induced dimerization can efficiently regulate Cre, and should be useful to achieve a tight temporal control of its activity, such as in the case of the creation of conditional knock-out animals. PMID:14576331

  1. Regulation of Cre recombinase by ligand-induced complementation of inactive fragments.

    PubMed

    Jullien, Nicolas; Sampieri, François; Enjalbert, Alain; Herman, Jean-Paul

    2003-11-01

    Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. To overcome this, we have developed DiCre, a regulatable fragment complementation system for Cre. The enzyme was split into two moieties that were fused to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12-rapamycin-associated protein), respectively. These can be efficiently heterodimerized by rapamycin. Several variants, based on splitting Cre at different sites and using different linker peptides, were tested in an indicator cell line. The fusion proteins, taken separately, had no recombinase activity. Stable transformants, co-expressing complementing fragments based on splitting Cre between Asn59 and Asn60, displayed low background activity affecting 0.05-0.4% of the cells. Rapamycin induced a rapid recombination, reaching 100% by 48-72 h, with an EC50 of 0.02 nM. Thus, ligand-induced dimerization can efficiently regulate Cre, and should be useful to achieve a tight temporal control of its activity, such as in the case of the creation of conditional knock-out animals.

  2. Evaluating Tissue-Specific Recombination in a Pdgfrα-CreERT2 Transgenic Mouse Line

    PubMed Central

    O’Rourke, Megan; Cullen, Carlie L.; Auderset, Loic; Pitman, Kimberley A.; Achatz, Daniela; Gasperini, Robert; Young, Kaylene M.

    2016-01-01

    In the central nervous system (CNS) platelet derived growth factor receptor alpha (PDGFRα) is expressed exclusively by oligodendrocyte progenitor cells (OPCs), making the Pdgfrα promoter an ideal tool for directing transgene expression in this cell type. Two Pdgfrα-CreERT2 mouse lines have been generated for this purpose which, when crossed with cre-sensitive reporter mice, allow the temporally restricted labelling of OPCs for lineage-tracing studies. These mice have also been used to achieve the deletion of CNS-specific genes from OPCs. However the ability of Pdgfrα-CreERT2 mice to induce cre-mediated recombination in PDGFRα+ cell populations located outside of the CNS has not been examined. Herein we quantify the proportion of PDGFRα+ cells that become YFP-labelled following Tamoxifen administration to adult Pdgfrα-CreERT2::Rosa26-YFP transgenic mice. We report that the vast majority (>90%) of PDGFRα+ OPCs in the CNS, and a significant proportion of PDGFRα+ stromal cells within the bone marrow (~38%) undergo recombination and become YFP-labelled. However, only a small proportion of the PDGFRα+ cell populations found in the sciatic nerve, adrenal gland, pituitary gland, heart, gastrocnemius muscle, kidney, lung, liver or intestine become YFP-labelled. These data suggest that Pdgfrα-CreERT2 transgenic mice can be used to achieve robust recombination in OPCs, while having a minimal effect on most PDGFRα+ cell populations outside of the CNS. PMID:27626928

  3. Targeted viral delivery of Cre recombinase induces conditional gene deletion in cardiovascular circuits of the mouse brain.

    PubMed

    Sinnayah, Puspha; Lindley, Timothy E; Staber, Patrick D; Davidson, Beverly L; Cassell, Martin D; Davisson, Robin L

    2004-06-17

    The Cre/loxP system has shown promise for investigating genes involved in nervous system function and pathology, although its application for studying central neural regulation of cardiovascular function and disease has not been explored. Here, we report for the first time that recombination of loxP-flanked genes can be achieved in discrete cardiovascular regulatory nuclei of adult mouse brain using targeted delivery of adenovirus (Ad) or feline immunodeficiency virus (FIV) bearing Cre recombinase (Ad-Cre, FIV-Cre). Single stereotaxic microinjections of Ad-Cre or FIV-Cre into specific nuclei along the subfornical organ-hypothalamic-hypophysial and brain stem-parabrachial axes resulted in robust and highly localized gene deletion as early as 7 days and for as long as 3 wk in a reporter mouse model in which Cre recombinase activates beta-galactosidase expression. An even greater selectivity in Cre-mediated gene deletion could be achieved in unique subpopulations of cells, such as vasopressin-synthesizing magnocellular neurons, by delivering Ad-Cre via retrograde transport. Moreover, Ad-Cre and FIV-Cre induced gene recombination in differential cell populations within these cardiovascular nuclei. FIV-Cre infection resulted in LacZ activation selectively in neurons, whereas both neuronal and glial cell types underwent gene recombination upon infection with Ad-Cre. These results establish the feasibility of using a combination of viral and Cre/loxP technologies to target specific cardiovascular nuclei in the brain for conditional gene modification and suggest the potential of this approach for determining the functional role of genes within these sites.

  4. Raising Awareness in Science Education for Women (RAISE-W)

    NASA Astrophysics Data System (ADS)

    Faherty, Jacqueline K.; Holford, M.

    2014-01-01

    Raising Awareness in Science Education for Women (RAISE-W) is a 501c non profit corporation whose mission is to aid in increasing and retaining the number of women - especially underrepresented females - engaged in scientific teaching and research. Initiated by a Protein Chemist and an Astronomer, our ultimate goal has been to develop informational tools and create innovative outreach programs for women across all STEM fields. At present RAISE-W is recruiting women at the undergraduate, graduate, and early career stages to participate in a unique, 1-year, executive coaching program modeled after those used in the business sector.

  5. The Strategic Communication Plan: Effective Communication for Strategic Leaders.

    DTIC Science & Technology

    2007-11-02

    PLAN? 5 WHO SHOULD HAVE A STRATEGIC COMMUNICATIONS PLAN? 7 SUCCESSFUL STRATEGIC PLANNING BEGINS WITH STRATEGIC THINKING . 8 HOW IS THE STRATEGIC...communicate internally and externally.9 SUCCESSFUL STRATEGIC PLANNING BEGINS WITH STRATEGIC THINKING A key objective of strategic thinking is to formulate and...vision should be captured in the strategic communication plan. Yet, in many organizations, prescribed strategic planning systems are the death knell

  6. Strategic forces

    SciTech Connect

    Not Available

    1990-10-01

    The Air Force now plans to retain the Minuteman II and III missile force through fiscal year 2008. Introduced about 25 years ago, these missiles have served as a nuclear deterrence for longer than initially envisioned. Over the extended lives of the systems, questions have arisen over their continued reliability and operational effectiveness, particularly the Minuteman II system. Limited flight testing, due to a shortage of test missiles, and reduced reliability caused by age-related deterioration of guidance computers and propulsion motors are two factors undermining confidence in the Minuteman II. GAO believes that the Minuteman II could be retired before 1998 as presently contemplated under an assumption of a Strategic Arms Reduction Talks agreement. An alternative would be to reinstate the Air Force's plans to replace deteriorated missile components and acquire the assets needed to resume flight testing at rates necessary to restore and sustain confidence in the system's performance through fiscal year 2008. However, on the basis of current test schedules, GAO is concerned that components to test the missile's warheads will be depleted by about 1999.

  7. Conditional Wwox Deletion in Mouse Mammary Gland by Means of Two Cre Recombinase Approaches

    PubMed Central

    Ferguson, Brent W.; Gao, Xinsheng; Kil, Hyunsuk; Lee, Jaeho; Benavides, Fernando; Abba, Martin C.; Aldaz, C. Marcelo

    2012-01-01

    Loss of WWOX expression has been reported in many different cancers including breast cancer. Elucidating the function of this gene in adult tissues has not been possible with full Wwox knockout models. Here we characterize the first conditional models of Wwox ablation in mouse mammary epithelium utilizing two transgenic lines expressing Cre recombinase, keratin 5-Cre (BK5-Cre) and MMTV-Cre. In the BK5-Cre model we observed very efficient Wwox ablation in KO mammary glands. However, BK5-Cre Wwox KO animals die prematurely for unknown reasons. In the MMTV-Cre model we observed significant ablation of Wwox in mammary epithelium with no effect on survival. In both of these models we found that Wwox deletion resulted in impaired mammary branching morphogenesis. We demonstrate that loss of Wwox is not carcinogenic in our KO models. Furthermore, no evidence of increase proliferation or development of premalignant lesions was observed. In none of the models did loss of a single Wwox allele (i.e. haploinsufficiency) have any observable phenotypic effect in mammary gland. To better understand the function of Wwox in the mammary gland, transcriptome profiling was performed. We observed that Wwox ablation results in the deregulation of genes involved in various cellular processes. We found that expression of the non-canonical Wnt ligand, Wnt5a, was significantly upregulated in Wwox KO mammary epithelium. Interestingly, we also determined that components of the Jak/Stat3 signaling pathway were upregulated in KO mice and this correlated with a very robust increase in phospho-Stat3 signaling, which warrants further testing. Even though the loss of Wwox expression in breast and other cancers is very well documented, our findings suggest that Wwox does not act as a classical tumor suppressor as previously thought. PMID:22574198

  8. Generation of an estrogen receptor beta-iCre knock-in mouse

    PubMed Central

    Cacioppo, Joseph A; Koo, Yongbum; Patrick Lin, Po-Ching; Osmulski, Sarah A; Ko, Chunjoo D; Ko, CheMyong

    2015-01-01

    A novel knock-in mouse that expresses codon-improved Cre recombinase (iCre) under regulation of the estrogen receptor beta (Esr2) promoter was developed for conditional deletion of genes and for the spatial and/or temporal localization of Esr2 expression. ESR2 is one of two classical nuclear estrogen receptors and displays a spatio-temporal expression pattern and functions that are different from the other estrogen receptor, ESR1. A cassette was constructed that contained iCre, a polyadenylation sequence, and a neomycin selection marker. This construct was used to insert iCre in front of the endogenous start codon of the Esr2 gene of a C57BL/6J embryonic stem cell line via homologous recombination. Resulting Esr2-iCre mice were bred with ROSA26-lacZ and Ai9-RFP reporter mice to visualize cells of functional iCre expression. Strong expression was observed in the ovary, the pituitary, the interstitium of the testes, the head and tail but not body of the epididymis, skeletal muscle, the coagulation gland (anterior prostate), the lung, and the preputial gland. Additional diffuse or patchy expression was observed in the cerebrum, the hypothalamus, the heart, the adrenal gland, the colon, the bladder, and the pads of the paws. Overall, Esr2-iCre mice will serve as a novel line for conditionally ablating genes in Esr2-expressing tissues, identifying novel Esr2-expressing cells, and differentiating the functions of ESR2 and ESR1. PMID:26663382

  9. Heme oxygenase-1 expression protects the heart from acute injury caused by inducible Cre recombinase.

    PubMed

    Hull, Travis D; Bolisetty, Subhashini; DeAlmeida, Angela C; Litovsky, Silvio H; Prabhu, Sumanth D; Agarwal, Anupam; George, James F

    2013-08-01

    The protective effect of heme oxygenase-1 (HO-1) expression in cardiovascular disease has been previously demonstrated using transgenic animal models in which HO-1 is constitutively overexpressed in the heart. However, the temporal requirements for protection by HO-1 induction relative to injury have not been investigated, but are essential to employ HO-1 as a therapeutic strategy in human cardiovascular disease states. Therefore, we generated mice with cardiac-specific, tamoxifen (TAM)-inducible overexpression of a human HO-1 (hHO-1) transgene (myosin heavy chain (MHC)-HO-1 mice) by breeding mice with cardiac-specific expression of a TAM-inducible Cre recombinase (MHC-Cre mice), with mice containing an hHO-1 transgene preceded by a floxed-stop signal. MHC-HO-1 mice overexpress HO-1 mRNA and the enzymatically active protein following TAM administration (40 mg/kg body weight on 2 consecutive days). In MHC-Cre controls, TAM administration leads to severe, acute cardiac toxicity, cardiomyocyte necrosis, and 80% mortality by day 3. This cardiac toxicity is accompanied by a significant increase in inflammatory cells in the heart that are predominantly neutrophils. In MHC-HO-1 mice, HO-1 overexpression ameliorates the depression of cardiac function and high mortality rate observed in MHC-Cre mice following TAM administration and attenuates cardiomyocyte necrosis and neutrophil infiltration. These results highlight that HO-1 induction is sufficient to prevent the depression of cardiac function observed in mice with TAM-inducible Cre recombinase expression by protecting the heart from necrosis and neutrophil infiltration. These findings are important because MHC-Cre mice are widely used in cardiovascular research despite the limitations imposed by Cre-induced cardiac toxicity, and also because inflammation is an important pathological component of many human cardiovascular diseases.

  10. In Vivo Performance and Properties of Tamoxifen Metabolites for CreERT2 Control

    PubMed Central

    Dolbois, Aymeric; Blazkova, Kristyna; Hess, Christopher; Low, Larry W. L.; Burger, Sibylle; Samson, Natasha; Carney, Tom J.; Bartunek, Petr; Nevado, Cristina

    2016-01-01

    Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. CreERT2 is efficiently activated in a concentration-dependent manner by the Tamoxifen metabolite trans-4-OH-Tamoxifen (trans-4-OHT). Reproducible and efficient Cre/lox experimentation is hindered by the gradual loss of CreERT2 induction potency upon prolonged storage of dissolved trans-4-OHT, which potentially results from gradual trans-to-cis isomerization or degradation. Here, we combined zebrafish CreERT2 recombination experiments and cell culture assays to document the gradual activity loss of trans-4-OHT and describe the alternative Tamoxifen metabolite Endoxifen as more stable alternative compound. Endoxifen retains potent activation upon prolonged storage (3 months), yet consistently induces half the ERT2 domain fusion activity compared to fresh trans-4-OHT. Using 1H-NMR analysis, we reveal that trans-4-OHT isomerization is undetectable upon prolonged storage in either DMSO or Ethanol, ruling out isomer transformation as cause for the gradual loss of trans-4-OHT activity. We further establish that both trans-4-OHT and Endoxifen are insensitive to light exposure under regular laboratory handling conditions. We attribute the gradual loss of trans-4-OHT potency to precipitation over time, and show that heating of aged trans-4-OHT aliquots reinstates their CreERT2 induction potential. Our data establish Endoxifen as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination experiments. PMID:27077909

  11. In Vivo Performance and Properties of Tamoxifen Metabolites for CreERT2 Control.

    PubMed

    Felker, Anastasia; Nieuwenhuize, Susan; Dolbois, Aymeric; Blazkova, Kristyna; Hess, Christopher; Low, Larry W L; Burger, Sibylle; Samson, Natasha; Carney, Tom J; Bartunek, Petr; Nevado, Cristina; Mosimann, Christian

    2016-01-01

    Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. CreERT2 is efficiently activated in a concentration-dependent manner by the Tamoxifen metabolite trans-4-OH-Tamoxifen (trans-4-OHT). Reproducible and efficient Cre/lox experimentation is hindered by the gradual loss of CreERT2 induction potency upon prolonged storage of dissolved trans-4-OHT, which potentially results from gradual trans-to-cis isomerization or degradation. Here, we combined zebrafish CreERT2 recombination experiments and cell culture assays to document the gradual activity loss of trans-4-OHT and describe the alternative Tamoxifen metabolite Endoxifen as more stable alternative compound. Endoxifen retains potent activation upon prolonged storage (3 months), yet consistently induces half the ERT2 domain fusion activity compared to fresh trans-4-OHT. Using 1H-NMR analysis, we reveal that trans-4-OHT isomerization is undetectable upon prolonged storage in either DMSO or Ethanol, ruling out isomer transformation as cause for the gradual loss of trans-4-OHT activity. We further establish that both trans-4-OHT and Endoxifen are insensitive to light exposure under regular laboratory handling conditions. We attribute the gradual loss of trans-4-OHT potency to precipitation over time, and show that heating of aged trans-4-OHT aliquots reinstates their CreERT2 induction potential. Our data establish Endoxifen as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination experiments.

  12. Molecular switch of Cre/loxP for radiation modulated gene therapy on hepatoma

    NASA Astrophysics Data System (ADS)

    Hsieh, Ya-Ju; Chen, Fu-Du; Wang, Fu Hui; Ke, Chien Chih; Wang, Hsin-Ell; Liu, Ren-Shyan

    2007-02-01

    For the purpose of enhancement of AFP promoter for the use of radiation modulated gene therapy for hepatocellular carcinoma (HCC), we combined hepatitis B virus (HBV) enhancer II with AFP promoter which shows the selectivity to the target cells to control the Cre/loxP system. Different gene constructs, pE4luc, pE4Tk, EIIAPA-Cre, E4CMV-STOP-Tk and chimeric promoters combined with HBV enhancer were constructed and transfected into HepG2, HeLa and NIH-3T3 cell lines. Cell experiments revealed that E4 enhancer responses to radiation best after 60 h irradiation at a dose range of 5-7 Gy in HepG2 stable clone. The EIIAPA promoter provided high specificity to hepatoma and activated the Cre downstream and removed the stop cassette only in hepatoma cells. After removal of the stop cassette, the E4 response to radiation could encode more Tk protein and kill more tumor cells. In summary, the chimeric EIIAPA promoter can stringently control the expression of Cre recombinase only in HCC. The radiation effect of the EIIAPA-Cre and E4CMV-STOP-Tk system shows promising results in terms of cell survival of HCC.

  13. Cre Fused with RVG Peptide Mediates Targeted Genome Editing in Mouse Brain Cells In Vivo

    PubMed Central

    Zou, Zhiyuan; Sun, Zhaolin; Li, Pan; Feng, Tao; Wu, Sen

    2016-01-01

    Cell penetrating peptides (CPPs) are short peptides that can pass through cell membranes. CPPs can facilitate the cellular entry of proteins, macromolecules, nanoparticles and drugs. RVG peptide (RVG hereinafter) is a 29-amino-acid CPP derived from a rabies virus glycoprotein that can cross the blood-brain barrier (BBB) and enter brain cells. However, whether RVG can be used for genome editing in the brain has not been reported. In this work, we combined RVG with Cre recombinase for bacterial expression. The purified RVG-Cre protein cut plasmids in vitro and traversed cell membranes in cultured Neuro2a cells. By tail vein-injecting RVG-Cre into Cre reporter mouse lines mTmG and Rosa26lacZ, we demonstrated that RVG-Cre could target brain cells and achieve targeted somatic genome editing in adult mice. This direct delivery of the gene-editing enzyme protein into mouse brains with RVG is much safer than plasmid- or viral-based methods, holding promise for further applications in the treatment of various brain diseases. PMID:27983648

  14. Isolation of Murine Bone Marrow Derived Mesenchymal Stem Cells using Twist2 Cre Transgenic Mice

    PubMed Central

    Liu, Yaling; Wang, Liping; Fatahi, Reza; Kronenberg, Mark; Kalajzic, Ivo; Rowe, David; Li, Yingcui; Maye, Peter

    2010-01-01

    While human bone marrow derived mesenchymal stem cells (BMSCs) are of great interest for their potential therapeutic value, its murine equivalent remains an important basic research model that can provide critical insights into the biology of this progenitor cell population. Here we present a novel transgenic strategy that allowed for the selective identification and isolation of murine BMSCs at the early stages of stromal cell culture. This strategy involved crossing Twist2 –Cre mice with Cre reporter mice such as Z/EG or Ai9, which express EGFP or Tomato fluorescent protein, respectively, upon Cre mediated excision of a stop sequence. Using this approach, we identified an adherent fluorescent protein+ cell population (T2C+) that is present during the earliest stages of colony formation and by day 5 of culture represents ~20% of the total cell population. Cell surface profiling by flow cytometry showed that T2C+ cells are highly positive for SCA1 and CD29 and negative for CD45, CD117, TIE2, and TER119. Isolation of T2C+ cells by FACS selected for a cell population with skeletal potential that can be directed to differentiate into osteoblasts, adipocytes, or chondrocytes. We also demonstrated in a calvarial bone defect model that T2C+ cells retain a strong efficacy for osteogenic repair and can support a hematopoietic environment. Collectively, these studies provide evidence that the Twist2-Cre x Cre reporter breeding strategy can be used to positively identify and isolate multipotent murine BMSCs. PMID:20673822

  15. Postnatal lineage mapping of follicular melanocytes with the Tyr::CreER(T) (2) transgene.

    PubMed

    Harris, Melissa L; Pavan, William J

    2013-03-01

    One of the main advantages of using inducible and conditional transgenes to study pigment cell biology is that they allow for genetic manipulation within melanocytes after roles in general neural crest or melanoblast development have been fulfilled. Specifically, we focus here on the ability of the Tyr::CreER(T) (2) transgenic line to alter genes within follicular melanocytes postnatally. Using the Gt(ROSA)26Sor(tm1sor) reporter allele, we present in detail the expression and activity of Tyr::CreER(T) (2) when induced during hair morphogenesis and adult hair cycling. We find that despite similarities in expression pattern to endogenous TYR, Tyr::CreER(T) (2) is effective at targeting both undifferentiated and differentiated melanocytes within the hair follicle. We also find that Tyr::CreER(T) (2) provides the highest levels of recombination when induced during the early phases of hair growth. In conclusion, the descriptions provided here will guide future analyses of gene function within the melanocyte system of the hair follicle when using this Tyr::CreER(T) (2) transgene.

  16. Evaluation of Cre Recombinase Delivery in Mammalian Cells using Baculovirus Infection

    PubMed Central

    Erbs, Eric; Pradhan, Amynah A.; Matifas, Audrey; Kieffer, Brigitte L.; Massotte, Dominique

    2013-01-01

    In vivo conditional knock-out of a protein is a method of choice to decipher its biological function. It can be achieved by encoding the cre-recombinase on a recombinant virus to exert spatio-temporal control of its expression and enzymatic activity and, subsequently, of the target gene deletion. Recombinant baculoviruses have been successfully used to express a wide range of proteins in insect cells. More recently, their potential to infect mammalian cells has been addressed but, so far, their ability to yield a conditional knock-out as a result of efficient in vivo cre-recombinase gene delivery has not been examined. Cre-recombinase fused to the green fluorescent protein was cloned under the control of the CAG promoter in a recombinant Autographa californica baculovirus expressing the vesicular stomatitis virus envelope G protein for increased mammalian cell infection. Gene delivery was evaluated in vitro in mammalian cells, neuroblastoma and mouse primary neuronal cultures as well as in vivo in the mouse brain. Infection with adeno-associated viruses encoding the cre-recombinase fused to the green fluorescent protein was performed as a positive control. Our results indicate that baculovirus infection leads to functional cre-recombinase expression in non-neuronal and neuroblastoma cell lines but not in mouse primary neuronal cultures or brain. PMID:23732834

  17. Targeted JAM-C deletion in germ cells by Spo11-controlled Cre recombinase.

    PubMed

    Pellegrini, Manuela; Claps, Giuseppina; Orlova, Valeria V; Barrios, Florencia; Dolci, Susanna; Geremia, Raffaele; Rossi, Pellegrino; Rossi, Gabriele; Arnold, Bernd; Chavakis, Triantafyllos; Feigenbaum, Lionel; Sharan, Shyam K; Nussenzweig, Andre

    2011-01-01

    Meiosis is a crucial process for the production of functional gametes. However, the biological significance of many genes expressed during the meiotic phase remains poorly understood, mainly because of the lethal phenotypes of the knockout mice. Functional analysis of such genes using the conditional knockout approach is hindered by the lack of suitable Cre transgenic lines. We describe here the generation of transgenic mice expressing Cre recombinase under the control of the meiotic Spo11 gene. Using LacZ-R26(loxP) and EYFP-R26(loxP) reporter mice, we show the specific expression and activity of Cre during meiosis in males and females. Spo11(Cre) mice were then crossed with floxed Nbs1 and JAM-C mice to produce conditional knockouts. A strong reduction of Nbs1 and JAM-C protein levels was found in the testis. Although Nbs1-deleted mice developed minor gonadal abnormalities, JAM-C-knockout mice showed a spermiogenetic arrest, as previously described for the null mice. These results provide strong evidence that Spo11(Cre) transgenic mice represent a powerful tool for deleting genes of interest specifically in meiotic and/or in postmeiotic germ cells.

  18. Establishment of a tamoxifen-inducible Cre-driver mouse strain for widespread and temporal genetic modification in adult mice.

    PubMed

    Ichise, Hirotake; Hori, Akiko; Shiozawa, Seiji; Kondo, Saki; Kanegae, Yumi; Saito, Izumu; Ichise, Taeko; Yoshida, Nobuaki

    2016-07-29

    Temporal genetic modification of mice using the ligand-inducible Cre/loxP system is an important technique that allows the bypass of embryonic lethal phenotypes and access to adult phenotypes. In this study, we generated a tamoxifen-inducible Cre-driver mouse strain for the purpose of widespread and temporal Cre recombination. The new line, named CM32, expresses the GFPneo-fusion gene in a wide variety of tissues before FLP recombination and tamoxifen-inducible Cre after FLP recombination. Using FLP-recombined CM32 mice (CM32Δ mice) and Cre reporter mouse lines, we evaluated the efficiency of Cre recombination with and without tamoxifen administration to adult mice, and found tamoxifen-dependent induction of Cre recombination in a variety of adult tissues. In addition, we demonstrated that conditional activation of an oncogene could be achieved in adults using CM32Δ mice. CM32Δ;T26 mice, which harbored a Cre recombination-driven, SV40 large T antigen-expressing transgene, were viable and fertile. No overt phenotype was found in the mice up to 3 months after birth. Although they displayed pineoblastomas (pinealoblastomas) and/or thymic enlargement due to background Cre recombination by 6 months after birth, they developed epidermal hyperplasia when administered tamoxifen. Collectively, our results suggest that the CM32Δ transgenic mouse line can be applied to the assessment of adult phenotypes in mice with loxP-flanked transgenes.

  19. Cre Recombinase Gene Transfer In Vitro and Detection of loxP-Dependent Recombination.

    PubMed

    Ohtsubo, Kazuaki; Marth, Jamey D

    2007-07-01

    INTRODUCTIONAltering the genome of intact cells and organisms by site-specific DNA recombination has become an important gene-transfer methodology. The inclusion of exogenous recombinase target sequences within transferred DNA segments allows subsequent modifications to previously altered genomic structure that increase the utility of gene transfer and enhance experimental design. In this protocol, correctly targeted mouse embryonic stem (ES) cell clones bearing the F[tkneo] allele (containing several loxP sites) are subjected to in vitro Cre gene transfer to generate ES cell subclones bearing either Type 1 (Δ) or Type 2 (F) alleles. Type 2 ES cells are used to generate chimeric mice that are then crossed to germ-line Cre-expressing mice, such as ZP3-Cre transgenic mates. The additional time needed to breed the mice (~2-3 mo) is typically less troublesome than the cost and effort of maintaining multiple clone-derived lines of mice.

  20. All-in-one construct for genome engineering using Cre-lox technology.

    PubMed

    Mariscal, Ana M; González-González, Luis; Querol, Enrique; Piñol, Jaume

    2016-06-01

    Mycoplasma genitalium is an appealing model of a minimal cell and synthetic biology study, and it was one of the first organisms whose genome was fully sequenced and chemically synthesized. Despite its usefulness as a model organism, many genetic tools well established for other microorganisms are not currently available in mycoplasmas. We have developed several vectors to adapt the Cre-lox technology for genome engineering in M. genitalium, providing an all-in-one construct that could be also useful to obtain unmarked genetic modifications in many other slow growing microorganisms. This construct contains a modified promoter sequence based in TetR system that exhibits an enhanced control on Cre recombinase expression, virtually abolishing the presence of this recombinase in the absence of inducer. This allows to introduce the Cre recombinase gene and the desired genetic modification in a single transformation step. In addition, this inducible promoter may be a very promising tool for a wide range of molecular applications.

  1. Six2creFrs2α knockout mice are a novel model of renal cystogenesis

    PubMed Central

    Puri, Pawan; Bushnell, Daniel; Schaefer, Caitlin M.; Bates, Carlton M.

    2016-01-01

    Six2cre-mediated deletion of Frs2α (Six2creFrs2αKO), a major fibroblast growth factor receptor (Fgfr) docking protein in mouse nephron progenitors results in perinatal renal hypoplasia; however, postnatal Six2creFrs2αKO kidneys develop cysts. We sought to determine the pathogenesis of Six2creFrs2αKO cyst formation. We performed histological assays, Western blots, and quantitative PCR (qPCR). While embryonic day (E) 18.5 Six2Frs2αKO kidneys were hypoplastic and not cystic, postnatal day (P) 7 mutants had proximal tubular-derived cysts that nearly replaced the renal parenchyma by P21. Mutants had high proximal tubular proliferation rates and interstitial fibrosis, similar to known polycystic kidney disease (PKD) models. Six2creFrs2αKO kidneys also had upregulation of Wnt/βcatenin signaling, macrophage infiltration and chemokine production (e.g. ectopic Ccl2 in non-dilated proximal tubules), and augmented hedgehog signaling, features also seen in other PKD models. We saw increased Gli1 (hedgehog readout) in postnatal Six2creFrs2αKO interstitium and ectopic sonic hedgehog (Shh) in subsets of non-dilated P7 mutant proximal tubules (likely driving the stromal Gli expression). As ectopic tubular Shh and Ccl2 expression is seen after acute kidney injury (AKI), we interrogated another bone fide AKI marker, Kim1 and noted ectopic expression in P7 non-dilated proximal tubules. These observations suggest that aberrantly activated “AKI” pathways may drive pathogenesis in PKD. PMID:27853247

  2. Overlapping ETS and CRE Motifs ((G/C)CGGAAGTGACGTCA) preferentially bound by GABPα and CREB proteins.

    PubMed

    Chatterjee, Raghunath; Zhao, Jianfei; He, Ximiao; Shlyakhtenko, Andrey; Mann, Ishminder; Waterfall, Joshua J; Meltzer, Paul; Sathyanarayana, B K; FitzGerald, Peter C; Vinson, Charles

    2012-10-01

    Previously, we identified 8-bps long DNA sequences (8-mers) that localize in human proximal promoters and grouped them into known transcription factor binding sites (TFBS). We now examine split 8-mers consisting of two 4-mers separated by 1-bp to 30-bps (X(4)-N(1-30)-X(4)) to identify pairs of TFBS that localize in proximal promoters at a precise distance. These include two overlapping TFBS: the ETS⇔ETS motif ((C/G)CCGGAAGCGGAA) and the ETS⇔CRE motif ((C/G)CGGAAGTGACGTCAC). The nucleotides in bold are part of both TFBS. Molecular modeling shows that the ETS⇔CRE motif can be bound simultaneously by both the ETS and the B-ZIP domains without protein-protein clashes. The electrophoretic mobility shift assay (EMSA) shows that the ETS protein GABPα and the B-ZIP protein CREB preferentially bind to the ETS⇔CRE motif only when the two TFBS overlap precisely. In contrast, the ETS domain of ETV5 and CREB interfere with each other for binding the ETS⇔CRE. The 11-mer (CGGAAGTGACG), the conserved part of the ETS⇔CRE motif, occurs 226 times in the human genome and 83% are in known regulatory regions. In vivo GABPα and CREB ChIP-seq peaks identified the ETS⇔CRE as the most enriched motif occurring in promoters of genes involved in mRNA processing, cellular catabolic processes, and stress response, suggesting that a specific class of genes is regulated by this composite motif.

  3. Plasma is a strategic resource.

    PubMed

    Strengers, Paul F W; Klein, Harvey G

    2016-12-01

    Plasma-derived medicinal products (PDMPs) such as immunoglobulins and clotting factors are listed by the World Health Organization as essential medicines. These and other PDMPs are crucial for the prophylaxis and treatment of patients with bleeding disorders, immune deficiencies, autoimmune and inflammatory diseases, and a variety of congenital deficiency disorders. While changes in clinical practice in developed countries have reduced the need for red blood cell transfusions thereby significantly reducing the collection volumes of whole blood and recovered plasma suitable for fractionation, the need for PDMPs worldwide continues to increase. The majority of plasma supplies for the manufacture of PDMPs is met by the US commercial plasma industry. However, geographic imbalance in the collection of plasma raises concerns that local disruptions of plasma supplies could result in regional and global shortages of essential PDMPs. Plasma, which fits the definition of a strategic resource, that is, "an economically important raw material which is subject to a higher risk of supply interruption," should be considered a strategic resource comparable to energy and drinking water. Plasma collections should be increased outside the United States, including in low- and middle-income countries. The need for capacity building in these countries is an essential part to strengthen quality plasma collection. This will require changes in national and regional policies. We advocate the need for the restoration of an equitable balance of the international plasma supply to reduce the risk of supply shortages worldwide. Strategic independence of plasma should be endorsed on a global level.

  4. Moving Beyond Strategic Planning to Strategic Thinking.

    ERIC Educational Resources Information Center

    Wolverton, Mimi; Gmelch, Walter

    1999-01-01

    Examines a moderately sized Washington school district's efforts to move beyond strategic planning as a segregated activity toward thinking strategically about long-term plans to govern both tactical operations and the district's future. Top management grew to recognize the legitimacy of multiple external and internal constituent claims. (25…

  5. The Cre/loxP system in Giardia lamblia: genetic manipulations in a binucleate tetraploid protozoan.

    PubMed

    Wampfler, Petra B; Faso, Carmen; Hehl, Adrian B

    2014-07-01

    The bacteriophage-derived Cre/loxP system is a valuable tool that has revolutionised genetic and cell biological research in many organisms. We implemented this system in the intestinal parasite Giardia lamblia, an evolutionarily diverged protozoan whose binucleate and tetraploid genome organisation severely limits the application of reverse genetic approaches. We show that Cre-recombinase is functionally expressed in G. lamblia and demonstrate "recycling" of selectable markers. Providing the means for more complex and versatile genetic modifications, this technique massively increases the scope of functional investigations in G. lamblia and other protozoa with similar limitations with respect to genetic manipulation.

  6. Raising Boys' Achievement in Schools.

    ERIC Educational Resources Information Center

    Bleach, Kevan, Ed.

    This book offers insights into the range of strategies and good practice being used to raise the achievement of boys. Case studies by school-based practitioners suggest ideas and measures to address the issue of achievement by boys. The contributions are: (1) "Why the Likely Lads Lag Behind" (Kevan Bleach); (2) "Helping Boys Do…

  7. Raising the Bar for Teaching

    ERIC Educational Resources Information Center

    Mehta, Jal; Doctor, Joe

    2013-01-01

    The past year has seen the emergence of a broad consensus on raising the standards for entering the teaching profession. The NEA, AFT, and Council of Chief State School Officers all have said they want higher entry standards. Such an exam would be modeled after other professions and is a potential game changer. If sufficiently rigorous, the exam…

  8. NASA Strategic Plan

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The aforementioned strategic decisions and the overarching direction for America's aeronautics and space program are addressed in the Strategic Plan. Our Strategic Plan is critical to our ability to meet the challenges of this new era and deliver a vibrant aeronautics and space program that strengthens and inspires the Nation. The Plan is our top-level strategy.

  9. Learning to think strategically.

    PubMed

    1994-01-01

    Strategic thinking focuses on issues that directly affect the ability of a family planning program to attract and retain clients. This issue of "The Family Planning Manager" outlines the five steps of strategic thinking in family planning administration: 1) define the organization's mission and strategic goals; 2) identify opportunities for improving quality, expanding access, and increasing demand; 3) evaluate each option in terms of its compatibility with the organization's goals; 4) select an option; and 5) transform strategies into action. Also included in this issue is a 20-question test designed to permit readers to assess their "strategic thinking quotient" and a list of sample questions to guide a strategic analysis.

  10. Comparison of efficiency between FLPe and Cre for recombinase-mediated cassette exchange in vitro and in adenovirus vector production.

    PubMed

    Takata, Yuki; Kondo, Saki; Goda, Naoki; Kanegae, Yumi; Saito, Izumu

    2011-07-01

    Cre and FLP recombinases mediate not only specific deletions and insertions, but also the recombinase-mediated cassette exchange (RMCE) reaction, which is used in cell biotechnology including ES cells and mouse genetics. However, comparison of efficiencies for Cre and FLP in RMCE has not been made. We here examined the detailed process of RMCE with Cre and FLP in vitro using mutant loxP 2272 and three mutant FRTs (FRT G, FRT H, and FRT F3) and then quantitatively compared the RMCE reactions in vitro. Interestingly, in the in vitro reactions, the RMCE efficiency of Cre reached a plateau level of approximately 5% and did not proceed further, whereas that of FLPe reached approximately 12-13%, showing that FLPe reached a higher level of efficiency than Cre possibly when they were supplied at a very high concentration. Moreover, we quantitatively compared the production efficiency of E1-deleted adenovirus vector using the RMCE method with Cre or FLP. The results showed that FLPe was again found more efficient than Cre in RMCE reaction. Thus, although Cre is considered more active than, or similar to, FLPe, it may not be necessarily true for RMCE reaction. Possible reasons explaining these results are discussed.

  11. Cre recombinase-mediated site-specific recombination between plant chromosomes.

    PubMed Central

    Qin, M; Bayley, C; Stockton, T; Ow, D W

    1994-01-01

    We report the use of the bacteriophage P1 Cre-lox system for generating conservative site-specific recombination between tobacco chromosomes. Two constructs, one containing a promoterless hygromycin-resistance gene preceded by a lox site (lox-hpt) and the other containing a cauliflower mosaic virus 35S promoter linked to a lox sequence and the cre coding region (35S-lox-cre), were introduced separately into tobacco plants. Crosses between plants harboring either construct produced plants with the two constructs situated on different chromosomes. Plants with recombination events were identified by selecting for hygromycin resistance, a phenotype expressed upon recombination. Molecular analysis showed that these recombination events occurred specifically at the lox sites and resulted in the reciprocal exchange of flanking host DNA. Progenies of these plants showed 67-100% cotransmission of the new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferential cosegregation of translocated chromosomes. These results illustrate that site-specific recombination systems can be useful tools for the large-scale manipulation of eukaryotic chromosomes in vivo. Images PMID:8127869

  12. Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting…

    PubMed Central

    Tsien, Joe Z.

    2016-01-01

    Defining and manipulating specific neurons in the brain has garnered enormous interest in recent years, because such an approach is now widely recognized as crucial for deepening our understanding of how the brain works. When I started exploring the Cre-loxP recombination for brain research in the early 1990s, it was written off as a dead-end project by a young fool. Yet over the past 20 years, Cre-lox recombination-mediated neurogenetics has emerged as one of the most powerful and versatile technology platforms for cell-specific gene knockouts, transgenic overexpression, Brainbow imaging, neural pathway tracing with retrovirus and CLARITY, chemical genetics, and optogenetics. Its popularity and greater utility in neuroscience research is also largely thanks to the NIH’s bold Blueprint for Neuroscience Research Initiative to launch several Cre-driver resource projects, as well as individual laboratories and private research organizations. With newly-discovered, genetically-encoded molecules that are capable of responding to sonar and magnetic stimulation, for sonogenetics or magnetogenetics, respectively, or detecting rapid voltage changes in neurons, Cre-lox neurogenetics will continue to aid brain research for years to come. PMID:26925095

  13. Coming of Age of the Family CReWS: A Ballet in Three Acts.

    ERIC Educational Resources Information Center

    Hayman, John L., Jr.; Clemson, Barry A.

    The Center for Cooperative Research with Schools (CReWS) contracted to develop a management information system (MIS) for the National Regional Resources Center of Pennsylvania (NRRCP). The system was originally intended to transform NRRCP data into management--oriented information to support decision-making within the agency. After a year's work,…

  14. Cre-dependent DNA recombination activates a STING-dependent innate immune response

    PubMed Central

    Pépin, Geneviève; Ferrand, Jonathan; Höning, Klara; Jayasekara, W. Samantha N.; Cain, Jason E.; Behlke, Mark A.; Gough, Daniel J.; G. Williams, Bryan R.; Hornung, Veit; Gantier, Michael P.

    2016-01-01

    Gene-recombinase technologies, such as Cre/loxP-mediated DNA recombination, are important tools in the study of gene function, but have potential side effects due to damaging activity on DNA. Here we show that DNA recombination by Cre instigates a robust antiviral response in mammalian cells, independent of legitimate loxP recombination. This is due to the recruitment of the cytosolic DNA sensor STING, concurrent with Cre-dependent DNA damage and the accumulation of cytoplasmic DNA. Importantly, we establish a direct interplay between this antiviral response and cell–cell interactions, indicating that low cell densities in vitro could be useful to help mitigate these effects of Cre. Taking into account the wide range of interferon stimulated genes that may be induced by the STING pathway, these results have broad implications in fields such as immunology, cancer biology, metabolism and stem cell research. Further, this study sets a precedent in the field of gene-engineering, possibly applicable to other enzymatic-based genome editing technologies. PMID:27166376

  15. Genome Editing via Mobile Group-II Introns and Cre/lox

    NASA Astrophysics Data System (ADS)

    Enyeart, P. E.; Perutka, J.; Dao, M.; Ellington, A. E.

    2010-04-01

    Mobile group-II introns and the Cre/lox systems are combined to allow large segments of DNA to be removed or transferred within/between bacterial genomes. Planned applications include metabolic optimization and development of novel dissimilatory metal-reducing bacteria.

  16. Evaluation of brilliance CRE agar for the detection of carbapenem-resistant gram-negative bacteria.

    PubMed

    Bracco, Silvia; Migliavacca, Roberta; Pini, Beatrice; Corbo, Nicoletta; Nucleo, Elisabetta; Brigante, Gioconda; Piazza, Aurora; Micheletti, Piero; Luzzaro, Francesco

    2013-04-01

    The aim of this work was to evaluate the performance of the new chromogenic medium BrillianceTM CREAgar (Thermo Fisher Scientific) for determining the limit of detection of carbapenem-resistant enterobacteria (CRE). A total of 70 clinical isolates were studied. Of these, 30 were well-characterized CRE, including Klebsiella pneumoniae strains producing KPC-, VIM-, and OXA-type enzymes, VIM-positive Enterobacter cloacae and Escherichia coli, NDM-positive E. coli, and enterobacterial isolates characterized by porin loss associated with ESBL production or AmpC hyperproduction. Ten carbapenem-resistant non-fermentative isolates were also included as well as 30 carbapenem-susceptible isolates. Carbapenem-resistant strains were inoculated at three different concentrations onto Brilliance CRE Agar (from 1.5x101 CFU/ml up to 1.5x104 CFU/ml) whereas carbapenem-susceptible isolates were inoculated at a concentration of 1.5x102 CFU/ml. The medium sustained the growth of carbapenem-resistant isolates, showing detection limits from 1.5x101 CFU/ml (in 31/40 cases) to 1.5x104 CFU/ml. No growth was observed with carbapenem-sensitive control strains. Our results indicate that the Brilliance CRE Agar allows the growth of carbapenem-resistant isolates with low detection limits and could represent a useful screening medium for both enterobacteria and non-fermentative Gram-negative strains resistant to carbapenems.

  17. Strategic Analysis Overview

    NASA Technical Reports Server (NTRS)

    Cirillo, William M.; Earle, Kevin D.; Goodliff, Kandyce E.; Reeves, J. D.; Stromgren, Chel; Andraschko, Mark R.; Merrill, R. Gabe

    2008-01-01

    NASA s Constellation Program employs a strategic analysis methodology in providing an integrated analysis capability of Lunar exploration scenarios and to support strategic decision-making regarding those scenarios. The strategic analysis methodology integrates the assessment of the major contributors to strategic objective satisfaction performance, affordability, and risk and captures the linkages and feedbacks between all three components. Strategic analysis supports strategic decision making by senior management through comparable analysis of alternative strategies, provision of a consistent set of high level value metrics, and the enabling of cost-benefit analysis. The tools developed to implement the strategic analysis methodology are not element design and sizing tools. Rather, these models evaluate strategic performance using predefined elements, imported into a library from expert-driven design/sizing tools or expert analysis. Specific components of the strategic analysis tool set include scenario definition, requirements generation, mission manifesting, scenario lifecycle costing, crew time analysis, objective satisfaction benefit, risk analysis, and probabilistic evaluation. Results from all components of strategic analysis are evaluated a set of pre-defined figures of merit (FOMs). These FOMs capture the high-level strategic characteristics of all scenarios and facilitate direct comparison of options. The strategic analysis methodology that is described in this paper has previously been applied to the Space Shuttle and International Space Station Programs and is now being used to support the development of the baseline Constellation Program lunar architecture. This paper will present an overview of the strategic analysis methodology and will present sample results from the application of the strategic analysis methodology to the Constellation Program lunar architecture.

  18. Defective bone repair in mast cell-deficient Cpa3Cre/+ mice

    PubMed Central

    Chan, Daniel; Samberg, Robert; Abou-Rjeili, Mira; Wong, Timothy H.; Li, Ailian; Feyerabend, Thorsten B.; Rodewald, Hans-Reimer; Henderson, Janet E.; Martineau, Paul A.

    2017-01-01

    In the adult skeleton, cells of the immune system interact with those of the skeleton during all phases of bone repair to influence the outcome. Mast cells are immune cells best known for their pathologic role in allergy, and may be involved in chronic inflammatory and fibrotic disorders. Potential roles for mast cells in tissue homeostasis, vascularization and repair remain enigmatic. Previous studies in combined mast cell- and Kit-deficient KitW-sh/W-sh mice (KitW-sh) implicated mast cells in bone repair but KitW-sh mice suffer from additional Kit-dependent hematopoietic and non- hematopoietic deficiencies that could have confounded the outcome. The goal of the current study was to compare bone repair in normal wild type (WT) and Cpa3Cre/+ mice, which lack mast cells in the absence of any other hematopoietic or non- hematopoietic deficiencies. Repair of a femoral window defect was characterized using micro CT imaging and histological analyses from the early inflammatory phase, through soft and hard callus formation, and finally the remodeling phase. The data indicate 1) mast cells appear in healing bone of WT mice but not Cpa3Cre/+ mice, beginning 14 days after surgery; 2) re-vascularization of repair tissue and deposition of mineralized bone was delayed and dis-organised in Cpa3Cre/+ mice compared with WT mice; 3) the defects in Cpa3Cre/+ mice were associated with little change in anabolic activity and biphasic alterations in osteoclast and macrophage activity. The outcome at 56 days postoperative was complete bridging of the defect in most WT mice and fibrous mal-union in most Cpa3Cre/+ mice. The results indicate that mast cells promote bone healing, possibly by recruiting vascular endothelial cells during the inflammatory phase and coordinating anabolic and catabolic activity during tissue remodeling. Taken together the data indicate that mast cells have a positive impact on bone repair. PMID:28350850

  19. Strategic Defense Initiative - strategic implications. Study project

    SciTech Connect

    Russell, T.L.

    1988-03-31

    In March 1983, during this address to the nation, President Reagan initiated a major shift in U.S. strategic policy, as he indicated his desire to move away from the condition of mutual vulnerability as the primary deterrent to nuclear war. The Strategic Defense Initiative (SDI), as the vehicle for this shift, has become the focus of debate over implications of a new strategic policy. This study seeks to examine the strategic relationship, and the role of arms control during a transition from an offensive-dominant strategy to a defensive-dominant strategy. The methodology for the study is to present arguments for and against SDI, as they relate to the issues of stability and arms control, evaluate their validity, draw conclusions, and provide recommendations that may enhance international security during a transition period of SDI.

  20. 5-Aminolaevulinate synthase gene promoter contains two cAMP-response element (CRE)-like sites that confer positive and negative responsiveness to CRE-binding protein (CREB).

    PubMed Central

    Giono, L E; Varone, C L; Cánepa, E T

    2001-01-01

    The first and rate-controlling step of the haem biosynthetic pathway in mammals and fungi is catalysed by the mitochondrial-matrix enzyme 5-aminolaevulinate synthase (ALAS). The purpose of this work was to explore the molecular mechanisms involved in the cAMP regulation of rat housekeeping ALAS gene expression. Thus we have examined the ALAS promoter for putative transcription-factor-binding sites that may regulate transcription in a cAMP-dependent protein kinase (PKA)-induced context. Applying both transient transfection assays with a chloramphenicol acetyltransferase reporter gene driven by progressive ALAS promoter deletions in HepG2, and electrophoresis mobility-shift assays we have identified two putative cAMP-response elements (CREs) at positions -38 and -142. Functional analysis showed that both CRE-like sites were necessary for complete PKA induction, but only one for basal expression. Co-transfection with a CRE-binding protein (CREB) expression vector increased PKA-mediated induction of ALAS promoter transcriptional activity. However, in the absence of co-transfected PKA, CREB worked as a specific repressor for ALAS promoter activity. A CREB mutant deficient in a PKA phosphorylation site was unable to induce expression of the ALAS gene but could inhibit non-stimulated promoter activity. Furthermore, a DNA-binding mutant of CREB did not interfere with ALAS promoter basal activity. Site-directed-mutagenesis studies showed that only the nearest element to the transcription start site was able to inhibit the activity of the promoter. Therefore, we conclude that CREB, through its binding to CRE-like sites, mediates the effect of cAMP on ALAS gene expression. Moreover, we propose that CREB could also act as a repressor of ALAS transcription, but is able to reverse its role after PKA activation. Dephosphorylated CREB would interfere in a spatial-disposition-dependent manner with the transcriptional machinery driving inhibition of gene expression. PMID:11139395

  1. Rax-CreERT2 knock-in mice: a tool for selective and conditional gene deletion in progenitor cells and radial glia of the retina and hypothalamus.

    PubMed

    Pak, Thomas; Yoo, Sooyeon; Miranda-Angulo, Ana L; Miranda-Angulo, Ana M; Wang, Hong; Blackshaw, Seth

    2014-01-01

    To study gene function in neural progenitors and radial glia of the retina and hypothalamus, we developed a Rax-CreERT2 mouse line in which a tamoxifen-inducible Cre recombinase is inserted into the endogenous Rax locus. By crossing Rax-CreER(T2) with the Cre-dependent Ai9 reporter line, we demonstrate that tamoxifen-induced Cre activity recapitulates the endogenous Rax mRNA expression pattern. During embryonic development, Cre recombinase activity in Rax-CreER(T2) is confined to retinal and hypothalamic progenitor cells, as well as progenitor cells of the posterior pituitary. At postnatal time points, selective Cre recombinase activity is seen in radial glial-like cell types in these organs--specifically Müller glia and tanycytes--as well as pituicytes. We anticipate that this line will prove useful for cell lineage analysis and investigation of gene function in the developing and mature retina, hypothalamus and pituitary.

  2. Cultivating strategic thinking skills.

    PubMed

    Shirey, Maria R

    2012-06-01

    This department highlights change management strategies that may be successful in strategically planning and executing organizational change initiatives. With the goal of presenting practical approaches helpful to nurse leaders advancing organizational change, content includes evidence-based projects, tools, and resources that mobilize and sustain organizational change initiatives. In this article, the author presents an overview of strategic leadership and offers approaches for cultivating strategic thinking skills.

  3. Generation of a KOR-Cre Knockin Mouse Strain to Study Cells Involved in Kappa Opioid Signaling

    PubMed Central

    Kuzirian, Marissa S.; Snyder, Lindsey M.; Matsushita, Megumi; Lee, Michael C.; Ferguson, Carolyn; Homanics, Gregg E.; Barth, Alison L.; Ross, Sarah E.

    2015-01-01

    The kappa opioid receptor (KOR) has numerous important roles in the nervous system including the modulation of mood, reward, pain, and itch. In addition, KOR is expressed in many non-neuronal tissues. However, the specific cell types that express KOR are poorly characterized. Here, we report the development of a KOR-Cre knockin allele, which provides genetic access to cells that express KOR. In this mouse, Cre recombinase (Cre) replaces the initial coding sequence of the Opkr1 gene (encoding the kappa opioid receptor). We demonstrate that the KOR-Cre allele mediates recombination by embryonic day 14.5 (E14.5). Within the brain, KOR-Cre shows expression in numerous areas including the cerebral cortex, nucleus accumbens and striatum. In addition, this allele is expressed in epithelium and throughout many regions of the body including the heart, lung, and liver. Finally, we reveal that KOR-Cre mediates recombination of a subset of bipolar and amacrine cells in the retina. Thus, the KOR-Cre mouse line is a valuable new tool for conditional gene manipulation to enable the study of KOR. PMID:26575788

  4. Analysis of GzmbCre as a Model System for Gene Deletion in the Natural Killer Cell Lineage.

    PubMed

    Xu, Yiying; Evaristo, Cesar; Alegre, Maria-Luisa; Gurbuxani, Sandeep; Kee, Barbara L

    2015-01-01

    The analysis of gene function in mature and activated natural killer cells has been hampered by the lack of model systems for Cre-mediated recombination in these cells. Here we have investigated the utility of GzmbCre for recombination of loxp sequences in these cells predicated on the observation that Gzmb mRNA is highly expressed in mature and activated natural killer cells. Using two different reporter strains we determined that gene function could be investigated in mature natural killer cells after GzmbCre mediated recombination in vitro in conditions that lead to natural killer cell activation such as in the cytokine combination of interleukin 2 and interleukin 12. We demonstrated the utility of this model by creating GzmbCre;Rosa26IKKbca mice in which Cre-mediated recombination resulted in expression of constitutively active IKKβ, which results in activation of the NFκB transcription factor. In vivo and in vitro activation of IKKβ in natural killer cells revealed that constitutive activation of this pathway leads to natural killer cell hyper-activation and altered morphology. As a caveat to the use of GzmbCre we found that this transgene can lead to recombination in all hematopoietic cells the extent of which varies with the particular loxp flanked allele under investigation. We conclude that GzmbCre can be used under some conditions to investigate gene function in mature and activated natural killer cells.

  5. Strategic planning for neuroradiologists.

    PubMed

    Berlin, Jonathan W; Lexa, Frank J

    2012-08-01

    Strategic planning is becoming essential to neuroradiology as the health care environment continues to emphasize cost efficiency, teamwork and collaboration. A strategic plan begins with a mission statement and vision of where the neuroradiology division would like to be in the near future. Formalized strategic planning frameworks, such as the strengths, weaknesses, opportunities and threats (SWOT), and the Balanced Scorecard frameworks, can help neuroradiology divisions determine their current position in the marketplace. Communication, delegation, and accountability in neuroradiology is essential in executing an effective strategic plan.

  6. Sandia Strategic Plan 1997

    SciTech Connect

    1997-12-01

    Sandia embarked on its first exercise in corporate strategic planning during the winter of 1989. The results of that effort were disseminated with the publication of Strategic Plan 1990. Four years later Sandia conducted their second major planning effort and published Strategic Plan 1994. Sandia`s 1994 planning effort linked very clearly to the Department of Energy`s first strategic plan, Fueling a Competitive Economy. It benefited as well from the leadership of Lockheed Martin Corporation, the management and operating contractor. Lockheed Martin`s corporate success is founded on visionary strategic planning and annual operational planning driven by customer requirements and technology opportunities. In 1996 Sandia conducted another major planning effort that resulted in the development of eight long-term Strategic Objectives. Strategic Plan 1997 differs from its predecessors in that the robust elements of previous efforts have been integrated into one comprehensive body. The changes implemented so far have helped establish a living strategic plan with a stronger business focus and with clear deployment throughout Sandia. The concept of a personal line of sight for all employees to this strategic plan and its objectives, goals, and annual milestones is becoming a reality.

  7. Trichoderma reesei CRE1-mediated Carbon Catabolite Repression in Re-sponse to Sophorose Through RNA Sequencing Analysis

    PubMed Central

    Antoniêto, Amanda Cristina Campos; de Paula, Renato Graciano; Castro, Lílian dos Santos; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2016-01-01

    Carbon catabolite repression (CCR) mediated by CRE1 in Trichoderma reesei emerged as a mechanism by which the fungus could adapt to new environments. In the presence of readily available carbon sources such as glucose, the fungus activates this mechanism and inhibits the production of cellulolytic complex enzymes to avoid unnecessary energy expenditure. CCR has been well described for the growth of T. reesei in cellulose and glucose, however, little is known about this process when the carbon source is sophorose, one of the most potent inducers of cellulase production. Thus, we performed high-throughput RNA sequencing to better understand CCR during cellulase formation in the presence of sophorose, by comparing the mutant ∆cre1 with its parental strain, QM9414. Of the 9129 genes present in the genome of T. reesei, 184 were upregulated and 344 downregulated in the mutant strain ∆cre1 compared to QM9414. Genes belonging to the CAZy database, and those encoding transcription factors and transporters are among the gene classes that were repressed by CRE1 in the presence of sophorose; most were possible indirectly regulated by CRE1. We also observed that CRE1 activity is carbon-dependent. A recent study from our group showed that in cellulose, CRE1 repress different groups of genes when compared to sophorose. CCR differences between these carbon sources may be due to the release of cellodextrins in the cellulose polymer, resulting in different targets of CRE1 in both carbon sources. These results contribute to a better understanding of CRE1-mediated CCR in T. reesei when glucose comes from a potent inducer of cellulase production such as sophorose, which could prove useful in improving cellulase production by the biotechnology sector. PMID:27226768

  8. Trichoderma reesei CRE1-mediated Carbon Catabolite Repression in Re-sponse to Sophorose Through RNA Sequencing Analysis.

    PubMed

    Antoniêto, Amanda Cristina Campos; de Paula, Renato Graciano; Castro, Lílian Dos Santos; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2016-04-01

    Carbon catabolite repression (CCR) mediated by CRE1 in Trichoderma reesei emerged as a mechanism by which the fungus could adapt to new environments. In the presence of readily available carbon sources such as glucose, the fungus activates this mechanism and inhibits the production of cellulolytic complex enzymes to avoid unnecessary energy expenditure. CCR has been well described for the growth of T. reesei in cellulose and glucose, however, little is known about this process when the carbon source is sophorose, one of the most potent inducers of cellulase production. Thus, we performed high-throughput RNA sequencing to better understand CCR during cellulase formation in the presence of sophorose, by comparing the mutant ∆cre1 with its parental strain, QM9414. Of the 9129 genes present in the genome of T. reesei, 184 were upregulated and 344 downregulated in the mutant strain ∆cre1 compared to QM9414. Genes belonging to the CAZy database, and those encoding transcription factors and transporters are among the gene classes that were repressed by CRE1 in the presence of sophorose; most were possible indirectly regulated by CRE1. We also observed that CRE1 activity is carbon-dependent. A recent study from our group showed that in cellulose, CRE1 repress different groups of genes when compared to sophorose. CCR differences between these carbon sources may be due to the release of cellodextrins in the cellulose polymer, resulting in different targets of CRE1 in both carbon sources. These results contribute to a better understanding of CRE1-mediated CCR in T. reesei when glucose comes from a potent inducer of cellulase production such as sophorose, which could prove useful in improving cellulase production by the biotechnology sector.

  9. Modular Cre/lox system and genetic therapeutics for colorectal cancer.

    PubMed

    Bordonaro, Michael

    2009-01-01

    The Cre/lox system is a powerful tool for targeting therapeutic effectors in a wide variety of human disorders. I review a Cre/lox Wnt-targeted system that has shown promise against Wnt-positive colorectal cancer cell lines. In addition to Wnt-specific targeting of cell death inducers, the modular nature of this gene therapy model system can be exploited by designing positive and negative feedback loops to either amplify or inhibit Wnt activity for experimental or therapeutic benefit. I discuss the structural components and performance parameters of the system, the implication of these findings with respect to cancer stem cells, as well as the general applicability of this system to any disorder characterized by differential gene expression. I also consider the issue of gene delivery as well as in vivo testing requirements necessary for the further characterization and development of this system.

  10. The Cre-loxP recombination-based reporter system for plant transcriptional expression studies.

    PubMed

    Shigaki, Toshiro; Vyzasatya, Ravindranadha R; Sivitz, Ali B; Ward, John M; Sze, Heven; Hirschi, Kendal D

    2005-05-01

    To facilitate the characterization of plant genes, the Cre-loxP site-specific recombination system was adapted to make reporter vectors for plant expression studies. This system allows promoter fragments to be cloned into a small vector (univector) and subsequently recombined in vitro with binary vectors containing different reporter genes precisely at near-perfect efficiency. We have constructed univector-adapted vectors with three reporters, beta-glucuronidase, luciferase, and green fluorescent protein, and a BASTA-resistance gene for selection of plant transformants. Expression in plants using the new system was validated by comparison to conventional reporter vectors. These new vectors are efficient and economical alternatives to the other plant reporter vectors currently available. The royalty-free Cre-loxP system serves as a platform for the future expansion of recombination-based cloning vectors for plant research.

  11. Genetic targeting of the endoderm with claudin-6CreER

    PubMed Central

    Anderson, William J; Zhou, Qiao; Alcalde, Victor; Kaneko, Osamu F; Blank, Leah J; Sherwood, Richard I; Guseh, J Sawalla; Rajagopal, Jayaraj; Melton, Douglas A

    2008-01-01

    A full description of the ontogeny of the β cell would guide efforts to generate β cells from embryonic stem cells (ESCs). The first step requires an understanding of definitive endoderm: the genes and signals responsible for its specification, proliferation, and patterning. This report describes a global marker of definitive endoderm, Claudin-6 (Cldn6). We report its expression in early development with particular attention to definitive endoderm derivatives. To create a genetic system to drive gene expression throughout the definitive endoderm with both spatial and temporal control, we target the endogenous locus with an inducible Cre recombinase (Cre-ERT2) cassette. Cldn6 null mice are viable and fertile with no obvious phenotypic abnormalities. We also report a lineage analysis of the fate of Cldn6-expressing embryonic cells, which is relevant to the development of the pancreas, lung, and liver. PMID:18213590

  12. Strategic renewal for business units.

    PubMed

    Whitney, J O

    1996-01-01

    Over the past decade, business units have increasingly taken the role of strategy formulation away from corporate headquarters. The change makes sense: business units are closer to customers, competitors, and costs. Nevertheless, business units can fail, just as headquarters once did, by losing their focus on the organization's priorities and capabilities. John Whitney--turnaround expert and professor of management at Columbia University--offers a method for refocusing companies that he calls the strategic-renewal process. The principles behind the process are straightforward, but its execution demands extensive data, rigorous analysis, and the judgment of key decision makers. However, when applied with diligence, it can produce a strategy that yields both growth and profit. To carry out the process, managers must analyze, one by one or in logical groupings, the company's customers, the products it sells, and the services it offers in light of three criteria: strategic importance, significance, and profitability. Does a given customer, product, or service mesh with the organization's goals? Is it significant in terms of current and future revenues? And is it truly profitable when all costs are care fully considered? Customers, products, and services that do not measure up, says the author, must be weeded out relentlessly. Although the process is a painstaking one, the article offers clear thinking on why-and how-to go about it. A series of exhibits takes managers through the questions they need to raise, and two matrices offer Whitney's concentrated wisdom on when to cultivate--and when to prune.

  13. Characterization of Channelrhodopsin and Archaerhodopsin in Cholinergic Neurons of Cre-Lox Transgenic Mice

    PubMed Central

    Hedrick, Tristan; Danskin, Bethanny; Larsen, Rylan S.; Ollerenshaw, Doug; Groblewski, Peter; Valley, Matthew; Olsen, Shawn; Waters, Jack

    2016-01-01

    The study of cholinergic signaling in the mammalian CNS has been greatly facilitated by the advent of mouse lines that permit the expression of reporter proteins, such as opsins, in cholinergic neurons. However, the expression of opsins could potentially perturb the physiology of opsin-expressing cholinergic neurons or mouse behavior. Indeed, the published literature includes examples of cellular and behavioral perturbations in preparations designed to drive expression of opsins in cholinergic neurons. Here we investigate expression of opsins, cellular physiology of cholinergic neurons and behavior in two mouse lines, in which channelrhodopsin-2 (ChR2) and archaerhodopsin (Arch) are expressed in cholinergic neurons using the Cre-lox system. The two mouse lines were generated by crossing ChAT-Cre mice with Cre-dependent reporter lines Ai32(ChR2-YFP) and Ai35(Arch-GFP). In most mice from these crosses, we observed expression of ChR2 and Arch in only cholinergic neurons in the basal forebrain and in other putative cholinergic neurons in the forebrain. In small numbers of mice, off-target expression occurred, in which fluorescence did not appear limited to cholinergic neurons. Whole-cell recordings from fluorescently-labeled basal forebrain neurons revealed that both proteins were functional, driving depolarization (ChR2) or hyperpolarization (Arch) upon illumination, with little effect on passive membrane properties, spiking pattern or spike waveform. Finally, performance on a behavioral discrimination task was comparable to that of wild-type mice. Our results indicate that ChAT-Cre x reporter line crosses provide a simple, effective resource for driving indicator and opsin expression in cholinergic neurons with few adverse consequences and are therefore an valuable resource for studying the cholinergic system. PMID:27243816

  14. Epidemiology of carbapenem resistant Enterobacteriaceae (CRE) during 2000-2012 in Asia

    PubMed Central

    Xu, Yanling; Gu, Bing; Huang, Mao; Liu, Haiyan; Xu, Ting; Xia, Wenying

    2015-01-01

    Background Over the past decade, the worldwide emergence of carbapenem resistance in Enterobacteriaceae has become a severe public health issue. This meta-analysis aims to describe the epidemiology of carbapenem resistant Enterobacteriaceae (CRE) during the years of 2000-2012 in Asian area. Methods PubMed and Embase databases were searched to identify the qualified papers. Random or fixed-effect model was used to deal with the data. Results Over all the 49 Asian countries (or regions), only 37.5% [19] of them contributed epidemiology data of CRE, and the rest ones provided either only case reports or no information at all. In Asia, the prevalence of CRE was still low during the study period with average resistance rates of 0.6% (95% CI, 0.6-0.8%, imipenem) and 0.9% (95% CI, 0.7-1.2%, meropenem). Resistance rates to imipenem and meropenem in Enterobacteriaceae exhibited stably escalating trend. Similar trend can also be observed among each Enterobacteriaceae genus, such as E. coli, Klebsiella spp. and Enterobacer spp. Klebsiella spp. accounted for the largest proportion among the isolates resistant to imipenem, and then followed by E. coli and Serratia. The rank order of resistance rates to imipenem among Enterobacteriaceae genus during the period of 2000-2012 was as follows: Serratia spp. (1.8%) > Proteus spp. (1.6%) > Klebsiella spp. (0.8%) = Citrobacter spp. (0.8%) > Enterobacer spp. (0.7%) > E. coli (0.2%). Conclusions Given the fact that the prevalence of CRE was increasing during the past decade, it is urgent for us to establish regional surveillance worldwide, carry out more effective antibiotic stewardship and infection control measures to prevent further spread of carbapenem resistance in Enterobacteriaceae. PMID:25922715

  15. Being a Learner Using Social Media in School: The Case of Space2cre8

    ERIC Educational Resources Information Center

    Vasbø, Kristin Beate; Silseth, Kenneth; Erstad, Ola

    2014-01-01

    The aim of this article is to gain knowledge about what it means to be a learner using social media in an educational setting. The article presents an ethnographic study of students in a multiethnic community in Oslo who participate in a social networking site called Space2cre8 (S28). In this article, we set out to explore the kind of space for…

  16. Studies of Prostate Tumor Development via Cre/LoxP Technology

    DTIC Science & Technology

    2002-05-01

    in our facility and used routinely in our transgenic experiments (Robles et al., 1996; Rodriguez Puebla et al., 2000). Crosses with Rosa 26 mice have...In the case of K5-Cre we used the strategy used routinely in our laboratory with primers for the Globin intron (Rodriguez Puebla et al. 2000). In the... Barrios RJ, Morton RA, Boyce BF, DeMayo FJ, Finegold MJ, Angelopoulou R, Rosen JM, Greenberg NM. Metastatic prostate cancer in a transgenic mouse. Cancer

  17. Unexpected Cartilage Phenotype in CD4-Cre-Conditional SOS-Deficient Mice

    PubMed Central

    Guittard, Geoffrey; Gallardo, Devorah L.; Li, Wenmei; Melis, Nicolas; Lui, Julian C.; Kortum, Robert L.; Shakarishvili, Nicholas G.; Huh, Sunmee; Baron, Jeffrey; Weigert, Roberto; Kramer, Joshua A.; Samelson, Lawrence E.; Sommers, Connie L.

    2017-01-01

    RAS signaling is central to many cellular processes and SOS proteins promote RAS activation. To investigate the role of SOS proteins in T cell biology, we crossed Sos1f/f Sos2−/− mice to CD4-Cre transgenic mice. We previously reported an effect of these mutations on T cell signaling and T cell migration. Unexpectedly, we observed nodules on the joints of greater than 90% of these mutant mice at 5 months of age, especially on the carpal joints. As the mice aged further, some also displayed joint stiffness, hind limb paralysis, and lameness. Histological analysis indicated that the abnormal growth in joints originated from dysplastic chondrocytes. Second harmonic generation imaging of the carpal nodules revealed that nodules were encased by rich collagen fibrous networks. Nodules formed in mice also deficient in RAG2, indicating that conventional T cells, which undergo rearrangement of the T cell antigen receptor, are not required for this phenotype. CD4-Cre expression in a subset of cells, either immune lineage cells (e.g., non-conventional T cells) or non-immune lineage cells (e.g., chondrocytes) likely mediates the dramatic phenotype observed in this study. Disruptions of genes in the RAS signaling pathway are especially likely to cause this phenotype. These results also serve as a cautionary tale to those intending to use CD4-Cre transgenic mice to specifically delete genes in conventional T cells. PMID:28386265

  18. The CRE-method: A technique of homeomorphic imaging in processing of seismic data

    NASA Astrophysics Data System (ADS)

    Steentoft, H.; Rabbel, W.

    Since the early sixties horizontal stacking is a standard technique in reflection seismics to increase the signal/noise ratio. The core is to add all signals which are reflected at the same subsurface point (common reflection point CRP, common reflecting element CRE). The resulting seismogram approximates a section of 'zero-offset-traces' representing identical positions of source and receiver for each trace, although the field traces are generally recorded at non-zero-offset. The Common-Midpoint (CMP) method has been the standard technique in seismic data processing. The presumptions are horizontal, plane layering and the knowledge of the spatial distribution of seismic velocities for the normal moveout (NMO) correction. In the case of dipping or irregular reflectors the CMP geometry implies 'reflector point dispersal' resulting in a loss of structural resolution. Attempts have been made to overcome the problems. Most prominent are 'prestack migration' and the 'dip moveout' correction. The Common-Reflecting-Element (CRE) method is a new stacking algorithm for multifold covered seismic reflection data preventing reflector point dispersal. It works without any model and without any presumptions of the layering or velocity distribution. Various aspects of the CRE method are discussed.

  19. CRE and SRE mediate LPA-induced CCN1 transcription in mouse aortic smooth muscle cells.

    PubMed

    Dou, Quanlin; Hao, Feng; Sun, Longsheng; Xu, Xuemin; Cui, Mei-Zhen

    2017-03-01

    Lysophosphatidic acid (LPA), one component of oxidized low-density lipoprotein (ox-LDL), is a potent bioactive phospholipid. Our recent data reveal that LPA induces matricellular protein CCN1 (also known as Cyr61) expression in aortic smooth muscle cells (SMCs) and that CCN1 bridges LPA and integrin signaling pathways leading to SMC migration. Whether and how LPA regulates the transcriptional machinery of the CCN1 gene are unknown. In this study, we found that LPA markedly induces CCN1 mRNA expression in SMCs. Using deleting mutation and reporter gene strategies, we demonstrated regions from -2038 to -1787 and from -101 to +63 of the CCN1 promoter contain the essential regulatory elements. The serum response element (SRE) and cyclic AMP-response element (CRE) are located in these regions. LPA induced time-dependent phosphorylation of serum response factor (SRF) and CRE-binding protein (CREB) in mouse SMCs. Luciferase assays of a series of deleted, mutated CCN1 promoter-reporter gene constructs and dominant negative construct revealed the distal SRE and the proximal CRE in the CCN1 promoter are required for LPA-induced CCN1 gene expression. Our results imply that elevated LPA levels may trigger SMC migration and exacerbate restenosis and atherosclerotic lesions through the induced CCN1, which communicates with a set of plasma membrane proteins and intracellular kinases.

  20. All-in-one construct for genome engineering using Cre-lox technology

    PubMed Central

    Mariscal, Ana M.; González-González, Luis; Querol, Enrique; Piñol, Jaume

    2016-01-01

    Mycoplasma genitalium is an appealing model of a minimal cell and synthetic biology study, and it was one of the first organisms whose genome was fully sequenced and chemically synthesized. Despite its usefulness as a model organism, many genetic tools well established for other microorganisms are not currently available in mycoplasmas. We have developed several vectors to adapt the Cre-lox technology for genome engineering in M. genitalium, providing an all-in-one construct that could be also useful to obtain unmarked genetic modifications in many other slow growing microorganisms. This construct contains a modified promoter sequence based in TetR system that exhibits an enhanced control on Cre recombinase expression, virtually abolishing the presence of this recombinase in the absence of inducer. This allows to introduce the Cre recombinase gene and the desired genetic modification in a single transformation step. In addition, this inducible promoter may be a very promising tool for a wide range of molecular applications. PMID:27084897

  1. CollagenVI-Cre mice: A new tool to target stromal cells in secondary lymphoid organs

    PubMed Central

    Prados, Alejandro; Kollias, George; Koliaraki, Vasiliki

    2016-01-01

    Stromal cells in secondary lymphoid organs (SLOs) are non-hematopoietic cells involved in the regulation of adaptive immune responses. Three major stromal populations have been identified in adult SLOs: fibroblastic reticular cells (FRCs), follicular dendritic cells (FDCs) and marginal reticular cells (MRCs). The properties of these individual populations are not clearly defined, mainly due to the lack of appropriate genetic tools, especially for MRCs. Here, we analyzed stromal cell targeting in SLOs from a transgenic mouse strain that expresses Cre recombinase under the CollagenVI promoter, using lineage tracing approaches. We show that these mice target specifically MRCs and FDCs, but not FRCs in Peyer’s patches and isolated lymphoid follicles in the intestine. In contrast, stromal cells in lymph nodes and the spleen do not express the transgene, which renders ColVI-cre mice ideal for the specific targeting of stromal cells in the gut-associated lymphoid tissue (GALT). This funding further supports the hypothesis of organ-specific stromal precursors in SLOs. Interestingly, in all tissues analyzed, there was also high specificity for perivascular cells, which have been proposed to act as FDC precursors. Taken together, ColVI-Cre mice are a useful new tool for the dissection of MRC- and FDC-specific functions and plasticity in the GALT. PMID:27604178

  2. Mapping the consequence of Notch1 proteolysis in vivo with NIP-CRE.

    PubMed

    Vooijs, Marc; Ong, Chin-Tong; Hadland, Brandon; Huppert, Stacey; Liu, Zhenyi; Korving, Jeroen; van den Born, Maaike; Stappenbeck, Thaddeus; Wu, Yumei; Clevers, Hans; Kopan, Raphael

    2007-02-01

    The four highly conserved Notch receptors receive short-range signals that control many biological processes during development and in adult vertebrate tissues. The involvement of Notch1 signaling in tissue self-renewal is less clear, however. We developed a novel genetic approach N(1)IP-CRE (Notch1 Intramembrane Proteolysis) to follow, at high resolution, the descendents of cells experiencing Notch1 activation in the mouse. By combining N(1)IP-CRE with loss-of-function analysis, Notch activation patterns were correlated with function during development, self-renewal and malignancy in selected tissues. Identification of many known functions of Notch1 throughout development validated the utility of this approach. Importantly, novel roles for Notch1 signaling were identified in heart, vasculature, retina and in the stem cell compartments of self-renewing epithelia. We find that the probability of Notch1 activation in different tissues does not always indicate a requirement for this receptor and that gradients of Notch1 activation are evident within one organ. These findings highlight an underappreciated layer of complexity of Notch signaling in vivo. Moreover, NIP-CRE represents a general strategy applicable for monitoring proteolysis-dependent signaling in vivo.

  3. Generation of eGFP and Cre knockin rats by CRISPR/Cas9.

    PubMed

    Ma, Yuanwu; Ma, Jing; Zhang, Xu; Chen, Wei; Yu, Lei; Lu, Yingdong; Bai, Lin; Shen, Bin; Huang, Xingxu; Zhang, Lianfeng

    2014-09-01

    The type II bacterial CRISPR/Cas [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas)] system is a very valuable genome engineering tool, which has been widely used in genome editing of a variety of organisms. Previously, we generated floxed alleles in rats by CRISPR/Cas9. Here, we successfully use a two-cut strategy with one circular vector, which contains the exogenous cDNAs with homology arm regions, in generating knockin rats at the Trdmt1, Nestin and Cck loci. The efficiency of CRISPR/Cas9-mediated knockin was up to 54%. Furthermore, by crossing the Nestin-Cre rat with the Dnmt3b floxed rat and Cck-Cre with the Dnmt1 floxed rat, we detected Cre/loxP-mediated recombination in the F1 generation of rats. We also show that the knockin alleles were germline transmitted. These results provided a simple and flexible engineering strategy for the establishment of knockin rats.

  4. The Cre/loxP recombination system for production of infectious mouse polyomavirus.

    PubMed

    Hron, Tomáš; Spanielová, Hana; Suchanová, Jiřina; Forstová, Jitka

    2013-09-01

    Murine polyomavirus mutants are frequently produced for experimental as well as therapy purposes. Commonly used methods for preparation of mutant viral genomes from recombinant vectors are laborious and give variable yields and quality. We describe an efficient and reproducible Cre/loxP-mediated recombination system that generates polyomavirus genomes from recombinant plasmid in vivo. We designed and constructed two variants of recombinant vectors containing the wild-type polyomavirus genome flanked by loxP homologous sites. The loxP sites were introduced either into the intronic region of early genes or between the two poly(A) signal sites of convergent transcriptional units. After cotransfection of the recombinant plasmids with the Cre-expressing vector into mouse 3T6 cells, we obtained infectious virus from the genome variant containing loxP site in the intronic region, but we failed to isolate any infectious virus from the viral genome containing loxP site between poly(A) signals. We show that the Cre/loxP-based method of polyomavirus production is simple, expedient, and reproducible and works with satisfactory efficiency.

  5. Strategic Leadership in Schools

    ERIC Educational Resources Information Center

    Williams, Henry S.; Johnson, Teryl L.

    2013-01-01

    Strategic leadership is built upon traits and actions that encompass the successful execution of all leadership styles. In a world that is rapidly changing, strategic leadership in schools guides school leader through assuring constant improvement process by anticipating future trends and planning for them and noting that plans must be flexible to…

  6. 11. Strategic planning.

    PubMed

    2014-05-01

    There are several types of planning processes and plans, including strategic, operational, tactical, and contingency. For this document, operational planning includes tactical planning. This chapter examines the strategic planning process and includes an introduction into disaster response plans. "A strategic plan is an outline of steps designed with the goals of the entire organisation as a whole in mind, rather than with the goals of specific divisions or departments". Strategic planning includes all measures taken to provide a broad picture of what must be achieved and in which order, including how to organise a system capable of achieving the overall goals. Strategic planning often is done pre-event, based on previous experience and expertise. The strategic planning for disasters converts needs into a strategic plan of action. Strategic plans detail the goals that must be achieved. The process of converting needs into plans has been deconstructed into its components and includes consideration of: (1) disaster response plans; (2) interventions underway or planned; (3) available resources; (4) current status vs. pre-event status; (5) history and experience of the planners; and (6) access to the affected population. These factors are tempered by the local: (a) geography; (b) climate; (c) culture; (d) safety; and (e) practicality. The planning process consumes resources (costs). All plans must be adapted to the actual conditions--things never happen exactly as planned.

  7. Strategic Investments Overview

    NASA Technical Reports Server (NTRS)

    Comstock, Doug

    2004-01-01

    This viewgraph presentation provides an overview of the organizational hierarchy for strategic management and strategic investments at NASA. The presentation also relates these topics to the budgets it submits to Congress, strategies for space exploration research and development, and systems analysis.

  8. Manage "Human Capital" Strategically

    ERIC Educational Resources Information Center

    Odden, Allan

    2011-01-01

    To strategically manage human capital in education means restructuring the entire human resource system so that schools not only recruit and retain smart and capable individuals, but also manage them in ways that support the strategic directions of the organization. These management practices must be aligned with a district's education improvement…

  9. Developing Strategic Readers

    ERIC Educational Resources Information Center

    Jones, Jennifer; Leahy, Susie

    2006-01-01

    As elementary students progress from "learning to read" toward "reading to learn," it is vital that they become strategic readers. A strategic reader is one who understands when and how to use a strategy in order to comprehend text. While some students use strategies naturally during the reading process, most students must be taught how to…

  10. Strategic Risk Assessment

    NASA Technical Reports Server (NTRS)

    Derleth, Jason; Lobia, Marcus

    2009-01-01

    This slide presentation provides an overview of the attempt to develop and demonstrate a methodology for the comparative assessment of risks across the entire portfolio of NASA projects and assets. It includes information about strategic risk identification, normalizing strategic risks, calculation of relative risk score, and implementation options.

  11. Keeping the Strategic Flame

    DTIC Science & Technology

    1997-01-01

    thinking strategically fifty years ago when he observed what nuclear weapons implied: “Thus far the chief purpose of our military establishment has...reached beyond available technol- ogy. Airmen in that day were thinking strategically and thus laid the founda- tions for American security policies

  12. Unintended targeting of Dmp1-Cre reveals a critical role for Bmpr1a signaling in the gastrointestinal mesenchyme of adult mice

    PubMed Central

    Lim, Joohyun; Burclaff, Joseph; He, Guangxu; Mills, Jason C; Long, Fanxin

    2017-01-01

    Cre/loxP technology has been widely used to study cell type-specific functions of genes. Proper interpretation of such data critically depends on a clear understanding of the tissue specificity of Cre expression. The Dmp1-Cre mouse, expressing Cre from a 14-kb DNA fragment of the mouse Dmp1 gene, has become a common tool for studying gene function in osteocytes, but the presumed cell specificity is yet to be fully established. By using the Ai9 reporter line that expresses a red fluorescent protein upon Cre recombination, we find that in 2-month-old mice, Dmp1-Cre targets not only osteocytes within the bone matrix but also osteoblasts on the bone surface and preosteoblasts at the metaphyseal chondro-osseous junction. In the bone marrow, Cre activity is evident in certain stromal cells adjacent to the blood vessels, but not in adipocytes. Outside the skeleton, Dmp1-Cre marks not only the skeletal muscle fibers, certain cells in the cerebellum and the hindbrain but also gastric and intestinal mesenchymal cells that express Pdgfra. Confirming the utility of Dmp1-Cre in the gastrointestinal mesenchyme, deletion of Bmpr1a with Dmp1-Cre causes numerous large polyps along the gastrointestinal tract, consistent with prior work involving inhibition of BMP signaling. Thus, caution needs to be exercised when using Dmp1-Cre because it targets not only the osteoblast lineage at an earlier stage than previously appreciated, but also a number of non-skeletal cell types. PMID:28163952

  13. California Community Colleges Technology II Strategic Plan, 2000-2005.

    ERIC Educational Resources Information Center

    California Community Colleges, Sacramento. Office of the Chancellor.

    This Strategic Plan aims to address the issues facing California Community Colleges (CCCs) due to new technology. The issues raised by the report include: (1) the pervasive and economically and socially necessary use of the Internet; (2) the digital divide; (3) the need for integrating technology into teaching and learning; (4) the impact of Tidal…

  14. GENSAT BAC Cre-recombinase driver lines to study the functional organization of cerebral cortical and basal ganglia circuits

    PubMed Central

    Gerfen, Charles R.; Paletzki, Ronald; Heintz, Nathaniel

    2013-01-01

    Summary Recent development of molecular genetic techniques are rapidly advancing understanding of the functional role of brain circuits in behavior. Critical to this approach is the ability to target specific neuron populations and circuits. The collection of over 250 BAC Cre-recombinase driver lines produced by the GENSAT project provides a resource for such studies. Here we provide characterization of GENSAT BAC-Cre driver lines with expression in specific neuroanatomical pathways within the cerebral cortex and basal ganglia. PMID:24360541

  15. Strategic Management or Strategic Planning for Defense?

    DTIC Science & Technology

    1989-02-01

    manage at the regional or CinC level with an appreciation of strategic planning and management concepts currently taught at business schools . Military...those not in uniform. Science, engineering, and business schools all suggest that their faculties have experience tours so that they can appreciate

  16. Site-specific modification of genome with cell-permeable Cre fusion protein in preimplantation mouse embryo

    SciTech Connect

    Kim, Kyoungmi; Kim, Hwain; Lee, Daekee

    2009-10-09

    Site-specific recombination (SSR) by Cre recombinase and its target sequence, loxP, is a valuable tool in genetic analysis of gene function. Recently, several studies reported successful application of Cre fusion protein containing protein transduction peptide for inducing gene modification in various mammalian cells including ES cell as well as in the whole animal. In this study, we show that a short incubation of preimplantation mouse embryos with purified cell-permeable Cre fusion protein results in efficient SSR. X-Gal staining of preimplantation embryos, heterozygous for Gtrosa26{sup tm1Sor}, revealed that treatment of 1-cell or 2-cell embryos with 3 {mu}M of Cre fusion protein for 2 h leads to Cre-mediated excision in 70-85% of embryos. We have examined the effect of the concentration of the Cre fusion protein and the duration of the treatment on embryonic development, established a condition for full term development and survival to adulthood, and demonstrated the germ line transmission of excised Gtrosa26 allele. Potential applications and advantages of the highly efficient technique described here are discussed.

  17. GABPα Binding to Overlapping ETS and CRE DNA Motifs Is Enhanced by CREB1: Custom DNA Microarrays.

    PubMed

    He, Ximiao; Syed, Khund Sayeed; Tillo, Desiree; Mann, Ishminder; Weirauch, Matthew T; Vinson, Charles

    2015-07-16

    To achieve proper spatiotemporal control of gene expression, transcription factors cooperatively assemble onto specific DNA sequences. The ETS domain protein monomer of GABPα and the B-ZIP domain protein dimer of CREB1 cooperatively bind DNA only when the ETS ((C)/GCGGAA GT: ) and CRE ( GT: GACGTCAC) motifs overlap precisely, producing the ETS↔CRE motif ((C)/GCGGAA GT: GACGTCAC). We designed a Protein Binding Microarray (PBM) with 60-bp DNAs containing four identical sectors, each with 177,440 features that explore the cooperative interactions between GABPα and CREB1 upon binding the ETS↔CRE motif. The DNA sequences include all 15-mers of the form (C)/GCGGA--CG-, the ETS↔CRE motif, and all single nucleotide polymorphisms (SNPs), and occurrences in the human and mouse genomes. CREB1 enhanced GABPα binding to the canonical ETS↔CRE motif CCGGAAGT two-fold, and up to 23-fold for several SNPs at the beginning and end of the ETS motif, which is suggestive of two separate and distinct allosteric mechanisms of cooperative binding. We show that the ETS-CRE array data can be used to identify regions likely cooperatively bound by GABPα and CREB1 in vivo, and demonstrate their ability to identify human genetic variants that might inhibit cooperative binding.

  18. Generation of a Tlx1(CreER-Venus) knock-in mouse strain for the study of spleen development.

    PubMed

    Nakahara, Ryo; Kawai, Yasuhiro; Oda, Akihisa; Nishimura, Miyuki; Murakami, Akikazu; Azuma, Takachika; Kaifu, Tomonori; Goitsuka, Ryo

    2014-11-01

    The spleen is a lymphoid organ that serves as a unique niche for immune reactions, extramedullary hematopoiesis, and the removal of aged erythrocytes from the circulation. While much is known about the immunological functions of the spleen, the mechanisms governing the development and organization of its stromal microenvironment remain poorly understood. Here we report the generation and analysis of a Tlx1(Cre) (ER) (-Venus) knock-in mouse strain engineered to simultaneously express tamoxifen-inducible CreER(T2) and Venus fluorescent protein under the control of regulatory elements of the Tlx1 gene, which encodes a transcription factor essential for spleen development. We demonstrated that Venus as well as CreER expression recapitulates endogenous Tlx1 transcription within the spleen microenvironment. When Tlx1(Cre) (ER) (-Venus) mice were crossed with the Cre-inducible reporter strain, Tlx1-expressing cells as well as their descendants were specifically labeled following tamoxifen administration. We also showed by cell lineage tracing that asplenia caused by Tlx1 deficiency is attributable to altered contribution of mesenchymal cells in the spleen anlage to the pancreatic mesenchyme. Thus, Tlx1(Cre) (ER) (-Venus) mice represent a new tool for lineage tracing and conditional gene manipulation of spleen mesenchymal cells, essential approaches for understanding the molecular mechanisms of spleen development.

  19. An Lck-cre transgene accelerates autoantibody production and lupus development in (NZB × NZW)F1 mice.

    PubMed

    Nelson, R K; Gould, K A

    2016-02-01

    Lupus is an autoimmune disease characterized by the development of antinuclear autoantibodies and immune complex-mediated tissue damage. T cells in lupus patients appear to undergo apoptosis at an increased rate, and this enhanced T cell apoptosis has been postulated to contribute to lupus pathogenesis by increasing autoantigen load. However, there is no direct evidence to support this hypothesis. In this study, we show that an Lck-cre transgene, which increases T cell apoptosis as a result of T cell-specific expression of cre recombinase, accelerates the development of autoantibodies and nephritis in lupus-prone (NZB × NZW)F1 mice. Although the enhanced T cell apoptosis in Lck-cre transgenic mice resulted in an overall decrease in the relative abundance of splenic CD4(+) and CD8(+) T cells, the proportion of activated CD4(+) T cells was increased and no significant change was observed in the relative abundance of suppressive T cells. We postulate that the Lck-cre transgene promoted lupus by enhancing T cell apoptosis, which, in conjunction with the impaired clearance of apoptotic cells in lupus-prone mice, increased the nuclear antigen load and accelerated the development of anti-nuclear autoantibodies. Furthermore, our results also underscore the importance of including cre-only controls in studies using the cre-lox system.

  20. Successful Community College Fund-Raising Programs

    ERIC Educational Resources Information Center

    Anderson, Spencer

    2005-01-01

    This article describes a study whose primary purposes were to determine the characteristics of an effective fund-raising program, the marketing practices that contribute to the success of a fund-raising program, and factors of the development system's influence on a fund-raising program. This study utilized a Delphi research instrument. Initially,…

  1. Fund-Raising by School Students.

    ERIC Educational Resources Information Center

    McClare, Greg; And Others

    1985-01-01

    A questionnaire completed by 138 principals indicated that fund-raising by pupils was extensive and was endorsed by parents, principals, and community organizations. The Toronto Board of Education subsequently adopted guidelines to ensure pupil safety in fund-raising outside school and to enhance the benefits of fund-raising, e.g., sharing,…

  2. Strategic nuclear forces of Britain and France

    SciTech Connect

    Prados, J.; Wit, J.S.; Zagurek, M.J. Jr.

    1984-08-01

    Weapon inventories of the US and the USSR account for about 97% of the nuclear warheads in the world today; Britain and France have most of the remainder. Yet even the comparatively modest claims the two Western European nations can make in this regard represent an awesome military capability that the USSR (and, to a lesser extent, the US) cannot afford to ignore. Indeed, the recent proposals both superpowers have made to dismantle all their European-based intermediate-range missiles differ sharply in particulars but raise the same fundamental question: What role will the other nuclear forces in Europe have in the future military balance. The Europeans are unlikely to place their limited strategic forces on the bargaining table along with those of the USSR and the US unless the superpowers impose severe constraints on their own strategic-weapon programs, and in particular on their strategic-defense programs. The deployment of such defense systems poses a worrisome dilemma for the Europeans: while in the near term a Soviet anti-ballistic-missile (ABM) system might have only marginal military impact, over the longer term a novel, exotic defense could conceivably nullify the dependent deterrent Britain and France have so arduously sought. At the same time and American system such as the Strategic Defense Initiative could, by decoupling the defense of Europe from that of the US, increase the need for precisely such an independent deterrent in the minds of the British and the French.

  3. American Strategic Minerals

    NASA Astrophysics Data System (ADS)

    DeYoung, John H., Jr.; Chidester, Alfred H.

    American Strategic Minerals is a collection of six papers that were presented in December 1982 at a conference organized by the Center for the Study of Marine Policy at the University of Delaware. According to editor Gerard J. Mangone, director of the center, the papers were commissioned “to investigate not only the objective resource situation, but also past United States policy on strategic minerals and future options open to Washington.” The authors and their chapter titles are John C. Kraft, University of Delaware: “Strategic minerals and world stability” V. Anthony Cammarota, Jr., U.S. Bureau of Mines: “America's dependence on strategic minerals” John D. Morgan, U.S. Bureau of Mines: “Future demands of the United States for strategic minerals” J. Robert Moore, University of Texas: “Alternative sources of strategic minerals from the seabed” Allan I. Mendelowitz and John E. Watson, U.S. General Accounting Office: “U.S. mining investments in developing countries” and James W. Curlin, Nautilus Press: “The political dimensions of strategic minerals.”

  4. Strategic Plan. Volume 1

    NASA Technical Reports Server (NTRS)

    2002-01-01

    The purpose of this document is to present the strategic plan and associated organizational structure that the National Space Biomedical Research Institute (NSBRI) will utilize to achieve the defined mission and objectives provided by NASA. Much of the information regarding the background and establishment of the NSBRI by NASA has been provided in other documentation and will not be repeated in this Strategic Plan. This Strategic Plan is presented in two volumes. Volume I (this volume) begins with an Introduction (Section 2) that provides the Institute's NASA-defined mission and objectives, and the organizational structure adopted to implement these through three Strategic Programs: Countermeasure Research; Education, Training and Outreach; and Cooperative Research and Development. These programs are described in Sections 3 to 5. Each program is presented in a similar way, using four subsections: Goals and Objectives; Current Strategies; Gaps and Modifications; and Resource Requirements. Section 6 provides the administrative infrastructure and total budget required to implement the Strategic Programs and assures that they form a single cohesive plan. This plan will ensure continued success of the Institute for the next five years. Volume II of the Strategic Plan provides an in-depth analysis of the current and future strategic programs of the 12 current NSBRI teams, including their goals, objectives, mutual interactions and schedules.

  5. The IAU Strategic Plan

    NASA Astrophysics Data System (ADS)

    Miley, George

    2016-10-01

    I shall review the content of the IAU Strategic Plan (SP) to use astronomy as a tool for stimulating development globally during the decade 2010 - 2020. Considerable progress has been made in its implementation since the last General Assembly.

  6. Engineering Forum Strategic Plan

    EPA Pesticide Factsheets

    This Strategic Plan highlights the purpose, mission, goals, and objectives of the U.S. Environmental Protection Agency (EPA) Engineering Forum (EF). It sets forth the principles that guide the EF's decision-making, helps clarify the EF's priorities, and...

  7. Information and strategic voting.

    PubMed

    Tyszler, Marcelo; Schram, Arthur

    2016-01-01

    We theoretically and experimentally study voter behavior in a setting characterized by plurality rule and mandatory voting. Voters choose from three options. We are interested in the occurrence of strategic voting in an environment where Condorcet cycles may occur and focus on how information about the preference distribution affects strategic behavior. We also vary the relative importance of the second preferred option. Quantal response equilibrium analysis is used to analyze the game and derive predictions. Our results indeed show that strategic voting arises. Its extent depends on (i) information availability; (ii) the relative importance of the intermediate candidate; (iii) the electorate's relative support for one's preferred candidate; (iv) the relative position of the plurality-supported candidate in one's preference ordering. Our results show that information serves as a coordination device where strategic voting does not harm the plurality-preferred candidate's chances of winning.

  8. USAF Strategic Master Plan

    DTIC Science & Technology

    2015-05-01

    objectives, and four annexes: the Human Capital Annex (HCA), Strategic Posture Annex (SPA), Capabilities Annex (CA), and the Science and Technology...64 ANNEX A - HUMAN CAPITAL ANNEX………………………………………………….………….A-1 ANNEX B - STRATEGIC...translate the SMP’s comprehensive goals and objectives into tangible actions and priorities. The four annexes are as follows:  Human Capital Annex (HCA

  9. 2015 Enterprise Strategic Vision

    SciTech Connect

    2015-08-01

    This document aligns with the Department of Energy Strategic Plan for 2014-2018 and provides a framework for integrating our missions and direction for pursuing DOE’s strategic goals. The vision is a guide to advancing world-class science and engineering, supporting our people, modernizing our infrastructure, and developing a management culture that operates a safe and secure enterprise in an efficient manner.

  10. Enhanced cell-permeant Cre protein for site-specific recombination in cultured cells

    PubMed Central

    Lin, Qing; Jo, Daewoong; Gebre-Amlak, Kassatihun D; Ruley, H Earl

    2004-01-01

    Background Cell-permeant Cre DNA site-specific recombinases provide an easily controlled means to regulate gene structure and function in living cells. Since recombination provides a stable and unambiguous record of protein uptake, the enzyme may also be used for quantitative studies of cis- and trans-acting factors that influence the delivery of proteins into cells. Results In the present study, 11 recombinant fusion proteins were analyzed to characterize sequences and conditions that affect protein uptake and/or activity and to develop more active cell-permeant enzymes. We report that the native enzyme has a low, but intrinsic ability to enter cells. The most active Cre proteins tested contained either an N-terminal 6xHis tag and a nuclear localization sequence from SV40 large T antigen (HNC) or the HIV Tat transduction sequence and a C-terminal 6xHis tag (TCH6). The NLS and 6xHis elements separately enhanced the delivery of the HNC protein into cells; moreover, transduction sequences from fibroblast growth factor 4, HIV Tat or consisting of the (KFF)3K sequence were not required for efficient protein transduction and adversely affected enzyme solubility. Transduction of the HNC protein required 10 to 15 min for half-maximum uptake, was greatly decreased at 4°C and was inhibited by serum. Efficient recombination was observed in all cell types tested (a T-cell line, NIH3T3, Cos7, murine ES cells, and primary splenocytes), and did not require localization of the enzyme to the nucleus. Conclusions The effects of different sequences on the delivery and/or activity of Cre in cultured cells could not be predicted in advance. Consequently, the process of developing more active cell-permeant recombinases was largely empirical. The HNC protein, with an excellent combination of activity, solubility and yield, will enhance the use of cell-permeant Cre proteins to regulate gene structure and function in living cells. PMID:15500682

  11. Atom probe field ion microscopy of Type 308 CRE stainless steel welds

    SciTech Connect

    Babu, S.S.; David, S.A.; Vitek, J.M.; Miller, M.K.

    1995-12-31

    Addition of controlled residual elements (CRE), such as 0.007 wt % B, to type 308 stainless steel welds, improved creep-rupture properties. In this paper, B distribution and microstructure development were studied. The microstructural evolution during high-temperature aging was found to similar to that of commercial SS308 welds. Atom probe analysis showed that B and C segregate to the ferrite-austenite interface. Thermodynamic calculations suggest that the segregation is due to preferential partitioning of B and C to the liquid during solidification. Further work is needed to study B redistribution in aging stages.

  12. The molecular biology of memory: cAMP, PKA, CRE, CREB-1, CREB-2, and CPEB.

    PubMed

    Kandel, Eric R

    2012-05-14

    The analysis of the contributions to synaptic plasticity and memory of cAMP, PKA, CRE, CREB-1, CREB-2, and CPEB has recruited the efforts of many laboratories all over the world. These are six key steps in the molecular biological delineation of short-term memory and its conversion to long-term memory for both implicit (procedural) and explicit (declarative) memory. I here first trace the background for the clinical and behavioral studies of implicit memory that made a molecular biology of memory storage possible, and then detail the discovery and early history of these six molecular steps and their roles in explicit memory.

  13. Strategic Stability Through the Strategic Defense Initiative

    DTIC Science & Technology

    1989-03-09

    role of the Strategic Defense Initiative research program will play in current and future nuclear stability issues. It addresses the historical ...Defense Initiative,.research program will play in current and future nuclear stability issues. It addresses the historical policies that lead to the...approach to stability, marked a historic moment. The question to consider from his speech is whether technological breakthroughs alter our earlier

  14. Strategic Forum. Number 274. January 2012. Raising Our Sights: Russian-American Strategic Restraint in an Age of Vulnerability

    DTIC Science & Technology

    2012-01-01

    of much higher U.S. defense priorities, such as in the Middle East and East Asia , and the need to cut defense spending.9 Yet despite the continued...weapons, instability and terrorist threats emanating from the Middle East and South Asia , disequi- librium in East Asia because of China’s rise. U.S... escala - tion in these new domains.20 Finally, NATO’s core principle of indivisible securi- ty requires the United States to insist that mutual strate- gic

  15. Guidelines for strategic planning

    SciTech Connect

    Not Available

    1991-07-01

    Strategic planning needs to be done as one of the integral steps in fulfilling our overall Departmental mission. The role of strategic planning is to assure that the longer term destinations, goals, and objectives which the programs and activities of the Department are striving towards are the best we can envision today so that our courses can then be set to move in those directions. Strategic planning will assist the Secretary, Deputy Secretary, and Under Secretary in setting the long-term directions and policies for the Department and in making final decisions on near-term priorities and resource allocations. It will assist program developers and implementors by providing the necessary guidance for multi-year program plans and budgets. It is one of the essential steps in the secretary's Strategic Planning Initiative. The operational planning most of us are so familiar with deals with how to get things done and with the resources needed (people, money, facilities, time) to carry out tasks. Operating plans like budgets, capital line item projects, R D budgets, project proposals, etc., are vital to the mission of the Department. They deal, however, with how to carry out programs to achieve some objective or budget assumption. Strategic planning deals with the prior question of what it is that should be attempted. It deals with what objectives the many programs and activities of the Department of Department should be striving toward. The purpose of this document is to provide guidance to those organizations and personnel starting the process for the first time as well as those who have prepared strategic plans in the past and now wish to review and update them. This guideline should not be constructed as a rigid, restrictive or confining rulebook. Each organization is encouraged to develop such enhancements as they think may be useful in their planning. The steps outlined in this document represent a very simplified approach to strategic planning. 9 refs.

  16. Strategic forces briefing

    SciTech Connect

    Bing, G.; Chrzanowski, P.; May, M.; Nordyke, M.

    1989-04-06

    The Strategic Forces Briefing'' is our attempt, accomplished over the past several months, to outline and highlight the more significant strategic force issues that must be addressed in the near future. Some issues are recurrent: the need for an effective modernized Triad and a constant concern for force survivability. Some issues derive from arms control: the Strategic Arms Reduction Talks (SALT) are sufficiently advanced to set broad numerical limits on forces, but not so constraining as to preclude choices among weapon systems and deployment modes. Finally, a new administration faced with serious budgetary problems must strive for the most effective strategic forces limited dollars can buy and support. A review of strategic forces logically begins with consideration of the missions the forces are charged with. We begin the briefing with a short review of targeting policy and implementation within the constraints of available unclassified information. We then review each element of the Triad with sections on SLBMs, ICBMs, and Air-Breathing (bomber and cruise missile) systems. A short section at the end deals with the potential impact of strategic defense on offensive force planning. We consider ABM, ASAT, and air defense; but we do not attempt to address the technical issues of strategic defense per se. The final section gives a brief overview of the tritium supply problem. We conclude with a summary of recommendations that emerge from our review. The results of calculation on the effectiveness of various weapon systems as a function of cost that are presented in the briefing are by Paul Chrzanowski.

  17. Using the Cre-lox system to randomize target gene expression states and generate diverse phenotypes

    PubMed Central

    Niesner, Bradley; Maheshri, Narendra

    2013-01-01

    Modifying the expression of multiple genes enables both deeper understanding of their function and the engineering of complex multigenic cellular phenotypes. However, deletion or overexpression of multiple genes can be laborious and involves multiple sequential genetic modifications. Here we describe a strategy to randomize the expression state of multiple genes in S. cerevisiae using Cre-loxP recombination. By inserting promoters flanked by inverted loxP sites in front of a gene of interest we can randomly turn it OFF or ON, or between 4 distinct expression states. We show at least 6 genes can be randomized independently and argue that using orthogonal loxP sites should increase this number to at least 15. Finally, we show how combining this strategy with mating allows easy introduction of native regulation as an additional expression state and use this to probe the role of 4 different enzymes involved in base excision repair in tolerate exposure to methyl methane sulfonate (MMS), a genotoxic DNA alkylating agent. The set of vectors developed here can be used to randomize the expression of both heterologous genes and endogenous genes, and could immediately prove useful for metabolic engineering in yeast. Because Cre-loxP recombination works in many organisms, this strategy should be readily extendable. PMID:23733452

  18. Comparison of Three Cre-LoxP Based Paired-End Library Construction Methods

    SciTech Connect

    Peng, Ze; Nath, Nandita; Tritt, Andrew; Liang, Shoudan; Han, James; Pennacchio, Len; Chen, Feng

    2013-03-26

    Paired-end library sequencing has been proven useful in scaffold construction during de novo whole genome shotgun assembly. The ability of generating mate pairs with > 8 Kb insert sizes is especially important for genomes containing long repeats. To make mate paired libraries for next generation sequencing, DNA fragments need to be circularized to bring the ends together. There are several methods that can be used for DNA circulation, namely ligation, hybridization and Cre-LoxP recombination. With higher circularization efficiency with large insert DNA fragments, Cre-LoxP recombination method generally has been used for constructing >8 kb insert size paired-end libraries. Second fragmentation step is also crucial for maintaining high library complexity and uniform genome coverage. Here we will describe the following three fragmentation methods: restriction enzyme digestion, random shearing and nick translation. We will present the comparison results for these three methods. Our data showed that all three methods are able to generate paired-end libraries with greater than 20 kb insert. Advantages and disadvantages of these three methods will be discussed as well.

  19. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain.

    PubMed

    Deverman, Benjamin E; Pravdo, Piers L; Simpson, Bryan P; Kumar, Sripriya Ravindra; Chan, Ken Y; Banerjee, Abhik; Wu, Wei-Li; Yang, Bin; Huber, Nina; Pasca, Sergiu P; Gradinaru, Viviana

    2016-02-01

    Recombinant adeno-associated viruses (rAAVs) are commonly used vehicles for in vivo gene transfer. However, the tropism repertoire of naturally occurring AAVs is limited, prompting a search for novel AAV capsids with desired characteristics. Here we describe a capsid selection method, called Cre recombination-based AAV targeted evolution (CREATE), that enables the development of AAV capsids that more efficiently transduce defined Cre-expressing cell populations in vivo. We use CREATE to generate AAV variants that efficiently and widely transduce the adult mouse central nervous system (CNS) after intravenous injection. One variant, AAV-PHP.B, transfers genes throughout the CNS with an efficiency that is at least 40-fold greater than that of the current standard, AAV9 (refs. 14,15,16,17), and transduces the majority of astrocytes and neurons across multiple CNS regions. In vitro, it transduces human neurons and astrocytes more efficiently than does AAV9, demonstrating the potential of CREATE to produce customized AAV vectors for biomedical applications.

  20. Neurogenin 3-directed cre deletion of Tsc1 gene causes pancreatic acinar carcinoma.

    PubMed

    Ding, Li; Han, Lingling; Li, Yin; Zhao, Jing; He, Ping; Zhang, Weizhen

    2014-11-01

    The role of tuberous sclerosis complex (TSC) in the pathogenesis of pancreatic cancers remains largely unknown. The present study shows that neurogenin 3 directed Cre deletion of Tsc1 gene induces the development of pancreatic acinar carcinoma. By cross-breeding the Neurog3-cre mice with Tsc1 (loxp/loxp) mice, we generated the Neurog3-Tsc1-/- transgenic mice in which Tsc1 gene is deleted and mTOR signaling activated in the pancreatic progenitor cells. All Neurog3-Tsc1-/- mice developed notable adenocarcinoma-like lesions in pancreas starting from the age of 100 days old. The tumor lesions are composed of cells with morphological and molecular resemblance to acinar cells. Metastasis of neoplasm to liver and lung was detected in 5% of animals. Inhibition of mTOR signaling by rapamycin significantly attenuated the growth of the neoplasm. Relapse of the neoplasm occurred within 14 days upon cessation of rapamycin treatment. Our studies indicate that activation of mTOR signaling in the pancreatic progenitor cells may trigger the development of acinar carcinoma. Thus, mTOR may serve as a potential target for treatment of pancreatic acinar carcinoma.

  1. Optimized second generation CRY2/CIB dimerizers and photoactivatable Cre recombinase

    PubMed Central

    Taslimi, Amir; Zoltowski, Brian; Miranda, Jose G.; Pathak, Gopal; Hughes, Robert M.; Tucker, Chandra L.

    2016-01-01

    Arabidopsis thaliana cryptochrome 2 (AtCRY2), a light-sensitive photosensory protein, was previously adapted for use controling protein-protein interactions through light-dependent binding to a partner protein, CIB1. While the existing CRY2/CIB dimerization system has been used extensively for optogenetic applications, some limitations exist. Here, we set out to optimize function of the CRY2/CIB system, to identify versions of CRY2/CIB that are smaller, show reduced dark interaction, and maintain longer or shorter signaling states in response to a pulse of light. We describe minimal functional CRY2 and CIB1 domains maintaining light-dependent interaction and new signaling mutations affecting AtCRY2 photocycle kinetics. The latter work implicates a α13-α14 turn motif within plant CRYs where perturbations alter signaling state lifetime. Using a long-lived L348F photocycle mutant, we engineered a second generation photoactivatable Cre recombinase, PA-Cre2.0, that shows five-fold improved dynamic range allowing robust recombination following exposure to a single, brief pulse of light. PMID:27065233

  2. Multidepartmental Response to a Duodenoscope Used on a CRE Patient: A Case Study.

    PubMed

    Bruns, Mark A; Gibbs, Elizabeth R

    Transmission of carbapenem-resistant Enterobacteriaceae (CRE) via duodenoscopes, specialized endoscopes used during endoscopic retrograde cholangiopancreatography (ERCP) procedures, has attracted media attention since early 2015. This attention has placed increasing focus on the reprocessing of duodenoscopes. Current reprocessing recommendations for these endoscopes require either high-level disinfection or ethylene oxide sterilization. While reprocessing duodenoscopes, staff at endoscopy locations within the Mercy health system perform a single high-level disinfection cycle that is preceded by two cycles of manual cleaning. The Mercy system has 37 locations for gastrointestinal endoscopic procedures and nine that can accommodate patients requiring ERCP. In early 2016, the Mercy Oklahoma City location performed an ERCP on a patient known prior to the case to be a carrier of CRE. After the case, multiple departments located in both the Oklahoma City and St. Louis facilities partnered to culture and sterilize the duodenoscope used in that case to ensure its safety for use on subsequent patients. This case study presents the situation and discusses culturing of endoscopes. In light of the evidence presented, the importance of enhanced communication and cooperation to achieve patient safety should be paramount to all other factors.

  3. Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration

    PubMed Central

    Feng, Dechun; Dai, Shen; Liu, Fengming; Ohtake, Yosuke; Zhou, Zhou; Wang, Hua; Zhang, Yonggang; Kearns, Alison; Peng, Xiao; Zhu, Faliang; Hayat, Umar; Li, Man; He, Yong; Xu, Mingjiang; Zhao, Chunling; Cheng, Min; Zhang, Lining; Wang, Hong; Yang, Xiaofeng; Ju, Cynthia; Bryda, Elizabeth C.; Gordon, Jennifer; Khalili, Kamel; Hu, Wenhui; Li, Shuxin; Qin, Xuebin

    2016-01-01

    Cell ablation is a powerful tool for studying cell lineage and/or function; however, current cell-ablation models have limitations. Intermedilysin (ILY), a cytolytic pore-forming toxin that is secreted by Streptococcus intermedius, lyses human cells exclusively by binding to the human complement regulator CD59 (hCD59), but does not react with CD59 from nonprimates. Here, we took advantage of this feature of ILY and developed a model of conditional and targeted cell ablation by generating floxed STOP-CD59 knockin mice (ihCD59), in which expression of human CD59 only occurs after Cre-mediated recombination. The administration of ILY to ihCD59+ mice crossed with various Cre-driver lines resulted in the rapid and specific ablation of immune, epithelial, or neural cells without off-target effects. ILY had a large pharmacological window, which allowed us to perform dose-dependent studies. Finally, the ILY/ihCD59-mediated cell-ablation method was tested in several disease models to study immune cell functionalities, hepatocyte and/or biliary epithelial damage and regeneration, and neural cell damage. Together, the results of this study demonstrate the utility of the ihCD59 mouse model for studying the effects of cell ablation in specific organ systems in a variety of developmental and disease states. PMID:27159394

  4. Nuclear DISC1 regulates CRE-mediated gene transcription and sleep homeostasis in the fruit fly

    PubMed Central

    Sawamura, Naoya; Ando, Tetsuya; Maruyama, Yasushi; Fujimuro, Masahiro; Mochizuki, Hiroaki; Honjo, Ken; Shimoda, Masami; Toda, Hirofumi; Sawamura-Yamamoto, Takako; Makuch, Lauren A; Hayashi, Akiko; Ishizuka, Koko; Cascella, Nicola G.; Kamiya, Atsushi; Ishida, Norio; Tomoda, Toshifumi; Hai, Tsonwin; Furukubo-Tokunaga, Katsuo; Sawa, Akira

    2009-01-01

    Disrupted-In-Schizophrenia-1 (DISC1) is one of major susceptibility factors for a wide range of mental illnesses, including schizophrenia, bipolar disorder, major depression, and autism spectrum conditions. DISC1 is located in several subcellular domains, such as the centrosome and the nucleus, and interacts with various proteins, including NudE-like (NUDEL/NDEL1) and activating transcription factor 4 (ATF4)/CREB2. Nevertheless, a role for DISC1 in vivo remains to be elucidated. Therefore, we have generated a Drosophila model for examining normal functions of DISC1 in living organisms. DISC1 transgenic flies with preferential accumulation of exogenous human DISC1 in the nucleus display disturbance in sleep homeostasis, which has been reportedly associated with CREB signaling/CRE-mediated gene transcription. Thus, in mammalian cells, we characterized nuclear DISC1, and identified a subset of nuclear DISC1 that co-localizes with the promyelocytic leukemia (PML) bodies, a nuclear compartment for gene transcription. Furthermore, we identified three functional cis-elements that regulate the nuclear localization of DISC1. We also report that DISC1 interacts with ATF4/CREB2 and a co-repressor N-CoR, modulating CRE-mediated gene transcription. PMID:18762802

  5. Nuclear DISC1 regulates CRE-mediated gene transcription and sleep homeostasis in the fruit fly.

    PubMed

    Sawamura, N; Ando, T; Maruyama, Y; Fujimuro, M; Mochizuki, H; Honjo, K; Shimoda, M; Toda, H; Sawamura-Yamamoto, T; Makuch, L A; Hayashi, A; Ishizuka, K; Cascella, N G; Kamiya, A; Ishida, N; Tomoda, T; Hai, T; Furukubo-Tokunaga, K; Sawa, A

    2008-12-01

    Disrupted-in-schizophrenia-1 (DISC1) is one of major susceptibility factors for a wide range of mental illnesses, including schizophrenia, bipolar disorder, major depression and autism spectrum conditions. DISC1 is located in several subcellular domains, such as the centrosome and the nucleus, and interacts with various proteins, including NudE-like (NUDEL/NDEL1) and activating transcription factor 4 (ATF4)/CREB2. Nevertheless, a role for DISC1 in vivo remains to be elucidated. Therefore, we have generated a Drosophila model for examining normal functions of DISC1 in living organisms. DISC1 transgenic flies with preferential accumulation of exogenous human DISC1 in the nucleus display disturbance in sleep homeostasis, which has been reportedly associated with CREB signaling/CRE-mediated gene transcription. Thus, in mammalian cells, we characterized nuclear DISC1, and identified a subset of nuclear DISC1 that colocalizes with the promyelocytic leukemia (PML) bodies, a nuclear compartment for gene transcription. Furthermore, we identified three functional cis-elements that regulate the nuclear localization of DISC1. We also report that DISC1 interacts with ATF4/CREB2 and a corepressor N-CoR, modulating CRE-mediated gene transcription.

  6. Flavonoids and Auxin Transport Inhibitors Rescue Symbiotic Nodulation in the Medicago truncatula Cytokinin Perception Mutant cre1

    PubMed Central

    Ng, Jason Liang Pin; Hassan, Samira; Truong, Thy T.; Hocart, Charles H.; Laffont, Carole; Frugier, Florian; Mathesius, Ulrike

    2015-01-01

    Initiation of symbiotic nodules in legumes requires cytokinin signaling, but its mechanism of action is largely unknown. Here, we tested whether the failure to initiate nodules in the Medicago truncatula cytokinin perception mutant cre1 (cytokinin response1) is due to its altered ability to regulate auxin transport, auxin accumulation, and induction of flavonoids. We found that in the cre1 mutant, symbiotic rhizobia cannot locally alter acro- and basipetal auxin transport during nodule initiation and that these mutants show reduced auxin (indole-3-acetic acid) accumulation and auxin responses compared with the wild type. Quantification of flavonoids, which can act as endogenous auxin transport inhibitors, showed a deficiency in the induction of free naringenin, isoliquiritigenin, quercetin, and hesperetin in cre1 roots compared with wild-type roots 24 h after inoculation with rhizobia. Coinoculation of roots with rhizobia and the flavonoids naringenin, isoliquiritigenin, and kaempferol, or with the synthetic auxin transport inhibitor 2,3,5,-triiodobenzoic acid, rescued nodulation efficiency in cre1 mutants and allowed auxin transport control in response to rhizobia. Our results suggest that CRE1-dependent cytokinin signaling leads to nodule initiation through the regulation of flavonoid accumulation required for local alteration of polar auxin transport and subsequent auxin accumulation in cortical cells during the early stages of nodulation. PMID:26253705

  7. Cloning-free genome engineering in Sinorhizobium meliloti advances applications of Cre/loxP site-specific recombination.

    PubMed

    Döhlemann, Johannes; Brennecke, Meike; Becker, Anke

    2016-09-10

    The soil-dwelling α-proteobacterium Sinorhizobium meliloti serves as model for studies of symbiotic nitrogen fixation, a highly important process in sustainable agriculture. Here, we report advancements of the genetic toolbox accelerating genome editing in S. meliloti. The hsdMSR operon encodes a type-I restriction-modification (R-M) system. Transformation of S. meliloti is counteracted by the restriction endonuclease HsdR degrading DNA which lacks the appropriate methylation pattern. We provide a stable S. meliloti hsdR deletion mutant showing enhanced transformation with Escherichia coli-derived plasmid DNA and demonstrate that using an E. coli plasmid donor, expressing S. meliloti methyl transferase genes, is an alternative strategy of increasing the transformation efficiency of S. meliloti. Furthermore, we devise a novel cloning-free genome editing (CFGE) method for S. meliloti, Agrobacterium tumefaciens and Xanthomonas campestris, and demonstrate the applicability of this method for intricate applications of the Cre/lox recombination system in S. meliloti. An enhanced Cre/lox system, allowing for serial deletions of large genomic regions, was established. An assay of lox spacer mutants identified a set of lox sites mediating specific recombination. The availability of several non-promiscuous Cre recognition sites enables simultaneous specific Cre/lox recombination events. CFGE combined with Cre/lox recombination is put forward as powerful approach for targeted genome editing, involving serial steps of manipulation to expedite the genetic accessibility of S. meliloti as chassis.

  8. Use of Cre/loxP recombination to swap cell binding motifs on the adenoviral capsid protein IX

    SciTech Connect

    Poulin, Kathy L.; Tong, Grace; Vorobyova, Olga; Pool, Madeline; Kothary, Rashmi; Parks, Robin J.

    2011-11-25

    We used Cre/loxP recombination to swap targeting ligands present on the adenoviral capsid protein IX (pIX). A loxP-flanked sequence encoding poly-lysine (pK-binds heparan sulfate proteoglycans) was engineered onto the 3'-terminus of pIX, and the resulting fusion protein allowed for routine virus propagation. Growth of this virus on Cre-expressing cells removed the pK coding sequence, generating virus that could only infect through alternative ligands, such as a tyrosine kinase receptor A (TrkA)-binding motif engineered into the capsid fibre protein for enhanced infection of neuronal cells. We used a similar approach to swap the pK motif on pIX for a sequence encoding a single-domain antibody directed towards CD66c for targeted infection of cancer cells; Cre-mediated removal of the pK-coding sequence simultaneously placed the single-domain antibody coding sequence in frame with pIX. Thus, we have developed a simple method to propagate virus lacking native viral tropism but containing cell-specific binding ligands. - Highlights: > We describe a method to grow virus lacking native tropism but containing novel cell-binding ligands. > Cre/loxP recombination was used to modify the adenovirus genome. > A targeting ligand present on capsid protein IX was removed or replaced using recombination. > Cre-loxP was also used to 'swap' the identity of the targeting ligand present on pIX.

  9. Flavonoids and Auxin Transport Inhibitors Rescue Symbiotic Nodulation in the Medicago truncatula Cytokinin Perception Mutant cre1.

    PubMed

    Ng, Jason Liang Pin; Hassan, Samira; Truong, Thy T; Hocart, Charles H; Laffont, Carole; Frugier, Florian; Mathesius, Ulrike

    2015-08-01

    Initiation of symbiotic nodules in legumes requires cytokinin signaling, but its mechanism of action is largely unknown. Here, we tested whether the failure to initiate nodules in the Medicago truncatula cytokinin perception mutant cre1 (cytokinin response1) is due to its altered ability to regulate auxin transport, auxin accumulation, and induction of flavonoids. We found that in the cre1 mutant, symbiotic rhizobia cannot locally alter acro- and basipetal auxin transport during nodule initiation and that these mutants show reduced auxin (indole-3-acetic acid) accumulation and auxin responses compared with the wild type. Quantification of flavonoids, which can act as endogenous auxin transport inhibitors, showed a deficiency in the induction of free naringenin, isoliquiritigenin, quercetin, and hesperetin in cre1 roots compared with wild-type roots 24 h after inoculation with rhizobia. Coinoculation of roots with rhizobia and the flavonoids naringenin, isoliquiritigenin, and kaempferol, or with the synthetic auxin transport inhibitor 2,3,5,-triiodobenzoic acid, rescued nodulation efficiency in cre1 mutants and allowed auxin transport control in response to rhizobia. Our results suggest that CRE1-dependent cytokinin signaling leads to nodule initiation through the regulation of flavonoid accumulation required for local alteration of polar auxin transport and subsequent auxin accumulation in cortical cells during the early stages of nodulation.

  10. Nerve growth factor enhances the CRE-dependent transcriptional activity activated by nobiletin in PC12 cells.

    PubMed

    Takito, Jiro; Kimura, Junko; Kajima, Koji; Uozumi, Nobuyuki; Watanabe, Makoto; Yokosuka, Akihito; Mimaki, Yoshihiro; Nakamura, Masanori; Ohizumi, Yasushi

    2016-07-01

    Prevention and treatment of Alzheimer disease are urgent problems for elderly people in developed countries. We previously reported that nobiletin, a poly-methoxylated flavone from the citrus peel, improved the symptoms in various types of animal models of memory loss and activated the cAMP responsive element (CRE)-dependent transcription in PC12 cells. Nobiletin activated the cAMP/PKA/MEK/Erk/MAPK signaling pathway without using the TrkA signaling activated by nerve growth factor (NGF). Here, we examined the effect of combination of nobiletin and NGF on the CRE-dependent transcription in PC12 cells. Although NGF alone had little effect on the CRE-dependent transcription, NGF markedly enhanced the CRE-dependent transcription induced by nobiletin. The NGF-induced enhancement was neutralized by a TrkA antagonist, K252a. This effect of NGF was effective on the early signaling event elicited by nobiletin. These results suggested that there was crosstalk between NGF and nobiletin signaling in activating the CRE-dependent transcription in PC12 cells.

  11. Children's Difficulty with Raising: A Performance Account

    ERIC Educational Resources Information Center

    Choe, Jinsun; Deen, Kamil

    2016-01-01

    This article explores English-speaking children's acquisition of raising structures with an experiencer (e.g.," John seems to Mary to be happy"). We review and address previously unnoticed issues in the methodologies of existing studies testing the acquisition of raising, thus providing a more reliable picture of children's abilities…

  12. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  13. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  14. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  15. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  16. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 2 2011-07-01 2011-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  17. Educational Fund Raising and the Law.

    ERIC Educational Resources Information Center

    Bargerstock, Charles T.

    The law relating to charitable gifts to schools is described, and fund raising considerations are addressed. The first of four chapters, "Education Fund Raising in the Legal Context," reviews the history of the law of charity over the past 400 years (i.e., law in England during 1500-1800 and law in America during 1800-1900). "The Donor and the…

  18. Adults raised as children in lesbian families.

    PubMed

    Tasker, F; Golombok, S

    1995-04-01

    A longitudinal study of 25 young adults from lesbian families and 21 raised by heterosexual single mothers revealed that those raised by lesbian mothers functioned well in adulthood in terms of psychological well-being and of family identity and relationships. The commonly held assumption that lesbian mothers will have lesbian daughters and gay sons was not supported by the findings.

  19. Raising Sensitive Issues in a Team

    ERIC Educational Resources Information Center

    Lindoerfer, Dennis

    2008-01-01

    Have you ever wondered how to deal with a sensitive issue within your team? For example, how do you raise the issue that the women rarely get listened to? How do you bring up your observation that the team members from Marketing always dominate the meetings? This guidebook focuses on ways to determine whether to raise such an issue in a team…

  20. NASA strategic plan

    NASA Technical Reports Server (NTRS)

    1994-01-01

    The NASA Strategic Plan is a living document. It provides far-reaching goals and objectives to create stability for NASA's efforts. The Plan presents NASA's top-level strategy: it articulates what NASA does and for whom; it differentiates between ends and means; it states where NASA is going and what NASA intends to do to get there. This Plan is not a budget document, nor does it present priorities for current or future programs. Rather, it establishes a framework for shaping NASA's activities and developing a balanced set of priorities across the Agency. Such priorities will then be reflected in the NASA budget. The document includes vision, mission, and goals; external environment; conceptual framework; strategic enterprises (Mission to Planet Earth, aeronautics, human exploration and development of space, scientific research, space technology, and synergy); strategic functions (transportation to space, space communications, human resources, and physical resources); values and operating principles; implementing strategy; and senior management team concurrence.

  1. Thinking strategically about capitation.

    PubMed

    Boland, P

    1997-05-01

    All managed care stakeholders--health plan members, employers, providers, community organizations, and government entitites--share a common interest in reducing healthcare costs while improving the quality of care health plan members receive. Although capitation is a usually thought of primarily as a payment mechanism, it can be a powerful tool providers and health plans can use to accomplish these strategic objectives and others, such as restoring and maintaining the health of plan members or improving a community's health status. For capitation to work effectively as a strategic tool, its use must be tied to a corporate agenda of partnering with stakeholders to achieve broader strategic goals. Health plans and providers must develop a partnership strategy in which each stakeholder has well-defined roles and responsibilities. The capitation structure must reinforce interdependence, shift focus from meeting organizational needs to meeting customer needs, and develop risk-driven care strategies.

  2. The Strategic Attitude: Integrating Strategic Planning into Daily University Worklife

    ERIC Educational Resources Information Center

    Dickmeyer, Nathan

    2004-01-01

    Chief financial officers in today's universities are so busy with the challenges of day-to-day management that strategic thinking often takes a back seat. Planning for strategic change can go a long way toward streamlining the very daily tasks that obscure the "big picture." Learning how to integrate strategic thinking into day-to-day management…

  3. Strategic Management in the Community College.

    ERIC Educational Resources Information Center

    Myran, Gunder A.

    1983-01-01

    Defines strategic management and discusses its role in community colleges, focusing on the components and methodology of strategic management, strategic and operational management function, management teams, and the need for strategic management. (DMM)

  4. On operator strategic behavior

    NASA Technical Reports Server (NTRS)

    Hancock, P. A.

    1991-01-01

    Deeper and more detailed knowledge as to how human operators such as pilots respond, singly and in groups, to demands on their performance which arise from technical systems will support the manipulation of such systems' design in order to accommodate the foibles of human behavior. Efforts to understand how self-autonomy impacts strategic behavior and such related issues as error generation/recognition/correction are still in their infancy. The present treatment offers both general and aviation-specific definitions of strategic behavior as precursors of prospective investigations.

  5. Strategic avionics technology planning

    NASA Technical Reports Server (NTRS)

    Cox, Kenneth J.; Brown, Don C.

    1991-01-01

    NASA experience in development and insertion of technology into programs had led to a recognition that a Strategic Plan for Avionics is needed for space. In the fall of 1989 an Avionics Technology Symposium was held in Williamsburg, Virginia. In early 1990, as a followon, a NASA wide Strategic Avionics Technology Working Group was chartered by NASA Headquarters. This paper will describe the objectives of this working group, technology bridging, and approaches to incentivize both the federal and commercial sectors to move toward rapidly developed, simple, and reliable systems with low life cycle cost.

  6. Strategic avionics technology planning

    NASA Astrophysics Data System (ADS)

    Cox, Kenneth J.; Brown, Don C.

    NASA experience in development and insertion of technology into programs had led to a recognition that a Strategic Plan for Avionics is needed for space. In the fall of 1989 an Avionics Technology Symposium was held in Williamsburg, Virginia. In early 1990, as a followon, a NASA wide Strategic Avionics Technology Working Group was chartered by NASA Headquarters. This paper will describe the objectives of this working group, technology bridging, and approaches to incentivize both the federal and commercial sectors to move toward rapidly developed, simple, and reliable systems with low life cycle cost.

  7. Microgravity strategic planning exercise

    NASA Technical Reports Server (NTRS)

    Halpern, Richard; Downey, Jim; Harvey, Harold

    1991-01-01

    The Center for Space and Advanced Technology supported a planning exercise for the Microgravity Program management at the Marshall Space Flight Center. The effort focused on the status of microgravity work at MSFC and elsewhere with the objective of preparing a goal-oriented strategic planning document which could be used for informational/brochure purposes. The effort entailed numerous interactions and presentations with Field Center programmatic components and Headquarters personnel. Appropriate material was consolidated in a draft format for a MSFC Strategic Plan.

  8. Tandem recombineering by SLIC cloning and Cre-LoxP fusion to generate multigene expression constructs for protein complex research.

    PubMed

    Haffke, Matthias; Viola, Cristina; Nie, Yan; Berger, Imre

    2013-01-01

    A robust protocol to generate recombinant DNA containing multigene expression cassettes by using sequence and ligation independent cloning (SLIC) followed by multiplasmid Cre-LoxP recombination in tandem for multiprotein complex research is described. The protocol includes polymerase chain reaction (PCR) amplification of the desired genes, seamless insertion into the target vector via SLIC, and Cre-LoxP recombination of specific donor and acceptor plasmid molecules, optionally in a robotic setup. This procedure, called tandem recombineering, has been implemented for multiprotein expression in E. coli and mammalian cells, and also for insect cells using a recombinant baculovirus.

  9. Comparison of the SUPERCARBA, CHROMagar KPC, and Brilliance CRE screening media for detection of Enterobacteriaceae with reduced susceptibility to carbapenems.

    PubMed

    Girlich, Delphine; Poirel, Laurent; Nordmann, Patrice

    2013-02-01

    The recently developed SUPERCARBA medium was evaluated together with 2 commercially available selective culture media containing carbapenems: CHROMagar KPC (CHROMagar) and Brilliance CRE (Oxoid, Thermofisher Scientific). A total of 142 enterobacterial isolates were tested, including 131 isolates with reduced susceptibility to carbapenems. The SUPERCARBA medium has the highest sensitivity (96.5%) (detecting virtually all carbapenemase producers including OXA-48-like producers) as compared to Brilliance CRE (76.3%) and CHROMagar KPC (43%). The specificity of the screening media was similar, ranging from 57% to 68%.

  10. Strategic plan for infrastructure optimization

    SciTech Connect

    Donley, C.D.

    1998-05-27

    This document represents Fluor Daniel Hanford`s and DynCorp`s Tri-Cities Strategic Plan for Fiscal Years 1998--2002, the road map that will guide them into the next century and their sixth year of providing safe and cost effective infrastructure services and support to the Department of Energy (DOE) and the Hanford Site. The Plan responds directly to the issues raised in the FDH/DOE Critical Self Assessment specifically: (1) a strategy in place to give DOE the management (systems) and physical infrastructure for the future; (2) dealing with the barriers that exist to making change; and (3) a plan to right-size the infrastructure and services, and reduce the cost of providing services. The Plan incorporates initiatives from several studies conducted in Fiscal Year 1997 to include: the Systems Functional Analysis, 200 Area Water Commercial Practices Plan, $ million Originated Cost Budget Achievement Plan, the 1OO Area Vacate Plan, the Railroad Shutdown Plan, as well as recommendations from the recently completed Review of Hanford Electrical Utility. These and other initiatives identified over the next five years will result in significant improvements in efficiency, allowing a greater portion of the infrastructure budget to be applied to Site cleanup. The Plan outlines a planning and management process that defines infrastructure services and structure by linking site technical base line data and customer requirements to work scope and resources. The Plan also provides a vision of where Site infrastructure is going and specific initiatives to get there.

  11. A Handbook for Strategic Planning

    DTIC Science & Technology

    1994-01-01

    This handbook was written for Department of the Navy (DON) commanding officers, TQL coordinators, and strategic planning facilitators in response to...questions about the strategic planning process and how it should be conducted within the DON. It is not intended to teach the intricacies of strategic ... planning , but is provided to answer process questions. While every question cannot be anticipated, the handbook details one way to do strategic

  12. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3....

  13. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3....

  14. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3....

  15. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3....

  16. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3....

  17. Strategic Planning and Financial Management

    ERIC Educational Resources Information Center

    Conneely, James F.

    2010-01-01

    Strong financial management is a strategy for strategic planning success in student affairs. It is crucial that student affairs professionals understand the necessity of linking their strategic planning with their financial management processes. An effective strategic planner needs strong financial management skills to implement the plan over…

  18. Strategic Partnerships in Higher Education

    ERIC Educational Resources Information Center

    Ortega, Janet L.

    2013-01-01

    The purpose of this study was to investigate the impacts of strategic partnerships between community colleges and key stakeholders; to specifically examine strategic partnerships; leadership decision-making; criteria to evaluate strategic partnerships that added value to the institution, value to the students, faculty, staff, and the local…

  19. Being Strategic in HE Management

    ERIC Educational Resources Information Center

    West, Andrew

    2008-01-01

    The call to be strategic--and with it the concept of strategic management--can bring to mind a wide range of definitions, and there is now a huge array of academic literature supporting the different schools of thought. At a basic level, however, strategic thinking is probably most simply about focusing on the whole, rather than the part. In…

  20. The Possibilities of Strategic Finance

    ERIC Educational Resources Information Center

    Chaffee, Ellen

    2010-01-01

    Strategic finance is aligning financial decisions--regarding revenues, creating and maintaining institutional assets, and using those assets--with the institution's mission and strategic plan. The concept known as "strategic finance" increasingly is being seen as a useful perspective for helping boards and presidents develop a sustainable…

  1. Linking Faculty Raises to College Resources

    ERIC Educational Resources Information Center

    Miller, John

    2006-01-01

    This article discusses how one small liberal arts school, Wheaton College in Massachusetts, devised a strategic plan to improve faculty salaries by linking improvements in salaries to improvements in college resources. The benefits of the salary plan went beyond improving salaries. First, the plan dictated that faculty salaries be determined at…

  2. Generation of ER{alpha}-floxed and knockout mice using the Cre/LoxP system

    SciTech Connect

    Antonson, P.; Omoto, Y.; Humire, P.; Gustafsson, J.-A.

    2012-08-10

    Highlights: Black-Right-Pointing-Pointer ER{alpha} floxed and knockout mice were generated. Black-Right-Pointing-Pointer Disruption of the ER{alpha} gene results in sterility in both male and female mice. Black-Right-Pointing-Pointer ER{alpha}{sup -/-} mice have ovaries with hemorrhagic follicles and hypoplastic uterus. Black-Right-Pointing-Pointer Female ER{alpha}{sup -/-} mice develop obesity. -- Abstract: Estrogen receptor alpha (ER{alpha}) is a nuclear receptor that regulates a range of physiological processes in response to estrogens. In order to study its biological role, we generated a floxed ER{alpha} mouse line that can be used to knock out ER{alpha} in selected tissues by using the Cre/LoxP system. In this study, we established a new ER{alpha} knockout mouse line by crossing the floxed ER{alpha} mice with Cre deleter mice. Here we show that genetic disruption of the ER{alpha} gene in all tissues results in sterility in both male and female mice. Histological examination of uterus and ovaries revealed a dramatically atrophic uterus and hemorrhagic cysts in the ovary. These results suggest that infertility in female mice is the result of functional defects of the reproductive tract. Moreover, female knockout mice are hyperglycemic, develop obesity and at the age of 4 months the body weight of these mice was more than 20% higher compared to wild type littermates and this difference increased over time. Our results demonstrate that ER{alpha} is necessary for reproductive tract development and has important functions as a regulator of metabolism in females.

  3. Transformation and Strategic Surprise

    DTIC Science & Technology

    2005-04-01

    not the enemy. Only in the rarest of cases is a strategic or operational level surprise itself so damaging that the defender is rendered incapable of...aptly perhaps a way with war, which has deprived its warriors, and the country, of political rewards earned and deserved by their blood . Given that our

  4. Strategic Sustainability Assessment

    DTIC Science & Technology

    2007-12-01

    complementary tools of forecasting and backcasting give regional stakeholders a unique perspective on potential policy and investment choices. Forecasting uses...and regional policy makers outside the fence. The project has several byproducts, for example, building beneficial partnerships with regional...transformations and their implications on the social, environmental, and economic fabric of the region. Backcasting is used to determine sets of strategic interven

  5. Strategic BI for All

    ERIC Educational Resources Information Center

    Raths, David

    2008-01-01

    Implementing a complex business intelligence (BI) system at a small school or one with limited resources can seem daunting. For small to midsize schools and community colleges, a strategic BI initiative may still be an elusive goal. This article discusses how schools with limited resources are making the dream a reality.

  6. Grappling with Strategic Dissonance.

    ERIC Educational Resources Information Center

    Dowie, Sandra

    2002-01-01

    Presents a case study of the Virtual Retina project (an instructional CD-ROM for ophthalmology students) at the University of Alberta as an example of strategic dissonance in an educational technology unit. Offers methods to analyze the external competitive environment and internal capabilities of educational technology units. (EV)

  7. The Strategic Assessment Model.

    ERIC Educational Resources Information Center

    Glazner, Steve, Ed.

    This book presents six papers focusing on the application of the strategic assessment model (SAM) to the management of higher education facilities. The papers are part of an ongoing effort by the Association of Higher Education Facilities Officers to provide comparative cost and staffing information and to develop a benchmarking process. The…

  8. External Strategic Planning Conference.

    ERIC Educational Resources Information Center

    Los Angeles Community Coll. District, CA. Office of Research and Planning.

    In response to a community reputation that has grown increasingly negative, the Los Angeles Community College District (LACCD) devised a strategic plan in 1998 to improve its programs and services and assure the educational success of its students. The planning process involved several steps: (1) revisiting the district mission statement; (2)…

  9. The Strategic Revolution.

    PubMed

    Gardner, Andy

    2016-09-08

    On the 40(th) anniversary of the publication of Richard Dawkins's The Selfish Gene, we explore the origins of cynical, strategic thinking in evolutionary biology, investigate how this illuminated the sexual and social lives of animals, and assess Dawkins's suggestion that evolution is best understood by taking the gene's-eye view.

  10. EMSL Strategic Plan 2008

    SciTech Connect

    Campbell, Allison A.

    2008-08-15

    This Strategic Plan is EMSL’s template for achieving our vision of simultaneous excellence in all aspects of our mission as a national scientific user facility. It reflects our understanding of the long-term stewardship we must work toward to meet the scientific challenges of the Department of Energy and the nation. During the next decade, we will implement the strategies contained in this Plan, working closely with the scientific community, our advisory committees, DOE’s Office of Biological and Environmental Research, and other key stakeholders. This Strategic Plan is fully aligned with the strategic plans of DOE and its Office of Science. We recognize that shifts in science and technology, national priorities, and resources made available through the Federal budget process create planning uncertainties and, ultimately, a highly dynamic planning environment. Accordingly, this Strategic Plan should be viewed as a living document for which we will continually evaluate changing needs and opportunities posed by our stakeholders (i.e., DOE, users, staff, advisory committees), work closely with them to understand and respond to those changes, and align our strategy accordingly.

  11. Strategic Education Research Partnership.

    ERIC Educational Resources Information Center

    Donovan, M. S., Ed.; Wigdor, A. K., Ed.; Snow, C. E., Ed.

    This book is a proposal for the Strategic Education Research Partnership (SERP), a large-scale coherent program of research and development carried out through a partnership between researchers and practitioners. The program would put the problems of educational practice at its center and focus on carrying research and development through all the…

  12. Strategic planning for marketers.

    PubMed

    Wilson, I

    1978-12-01

    The merits of strategic planning as a marketing tool are discussed in this article which takes the view that although marketers claim to be future-oriented, they focus too little attention on long-term planning and forecasting. Strategic planning, as defined by these authors, usually encompasses periods of between five and twenty-five years and places less emphasis on the past as an absolute predictor of the future. It takes a more probabilistic view of the future than conventional marketing strategy and looks at the corporation as but one component interacting with the total environment. Inputs are examined in terms of environmental, social, political, technological and economic importance. Because of its futuristic orientation, an important tenant of strategic planning is the preparation of several alternative scenarios ranging from most to least likely. By planning for a wide-range of future market conditions, a corporation is more able to be flexible by anticipating the course of future events, and is less likely to become a captive reactor--as the authors believe is now the case. An example of strategic planning at General Elecric is cited.

  13. Strategic Defense Initiative program

    SciTech Connect

    Conachan, F.C.

    1991-05-01

    This paper discusses the Strategic Defense Initiative (SDI) program. It summarizes the major programmatic and technological lessons learned from the SDI program over the past 7 years. It provides information on: past uses of SDI funds, persistent optimism in planning and starting projects, evolution of SDI architecture, and accuracy of cost estimates.

  14. Using Collaborative Strategic Reading.

    ERIC Educational Resources Information Center

    Klingner, Janette K.; Vaughn, Sharon

    1998-01-01

    Describes collaborative strategic reading (CSR), a technique for teaching students, such as those with learning disabilities, reading comprehension and vocabulary skills in a cooperative setting. Covers teaching the four strategies of CSR (preview, click and clunk, get the gist, and wrap up), as well as teaching students cooperative learning group…

  15. Measuring strategic success.

    PubMed

    Gish, Ryan

    2002-08-01

    Strategic triggers and metrics help healthcare providers achieve financial success. Metrics help assess progress toward long-term goals. Triggers signal market changes requiring a change in strategy. All metrics may not move in concert. Organizations need to identify indicators, monitor performance.

  16. Enduring Strategic Rivalries

    DTIC Science & Technology

    2014-08-01

    W. Harl, Tulane University ............................................................................... 4-1 “1066 and All That”: English and...Islamic invasion England vs. France (1066 to 1453, the Middle Ages) The 1066 Norman conquest of France English defeat that saw their military...1453) The creation of powerful centralized proto- states (as in Hundred Years’ War) Total English defeat 1-11 Introduction to Enduring Strategic

  17. Strategic Marketing for Agribusiness.

    ERIC Educational Resources Information Center

    Welch, Mary A., Ed.

    1993-01-01

    The steps for strategic market planning are discussed including: (1) assessing the situation with market conditions, customers, competitors, and your firm; and (2) crafting a strategy to prioritize target markets, develop a core strategy, and create a marketing mix. Examples of agribusiness successes are presented. The booklet concludes with a…

  18. Strategically Planning to Change

    ERIC Educational Resources Information Center

    Atkins, Kemal

    2010-01-01

    Higher education, like the private sector, is searching for innovative ways to respond to demographic shifts, globalization, greater accountability, and new technologies. New organizational models are needed to meet these challenges. In a rapidly changing world, the development of such models can occur through effective strategic analysis and…

  19. Cure therapeutics and strategic prevention: raising the bar for mental health research.

    PubMed

    Insel, T R; Scolnick, E M

    2006-01-01

    Mental disorders cause more disability than any other class of medical illness in Americans between ages 15 and 44 years. The suicide rate is higher than the annual mortality from homicide, AIDS, and most forms of cancer. In contrast to nearly all communicable and most non-communicable diseases, there is little evidence that the morbidity and mortality from mental disorders have changed in the past several decades. Mental health advocates, including psychiatric researchers, have pointed to stigma as one of the reasons for the lack of progress with mental illnesses relative to other medical illnesses. This review considers how the expectations and goals of the research community have contributed to this relative lack of progress. In contrast to researchers in cancer and heart disease who have sought cures and preventions, biological psychiatrists in both academia and industry have set their sights on incremental and marketable advances, such as drugs with fewer adverse effects. This essay argues for approaches that can lead to cures and strategies for prevention of schizophrenia and mood disorders.

  20. Product Fund-Raising: A Practical Resource.

    ERIC Educational Resources Information Center

    Lemieux, Russell A.

    1997-01-01

    Reviews critical issues in selecting a product fund-raising program for schools. Suggests that the most important considerations should be the quality of the product or service and its "fit" with the school. (JDI)

  1. Raise the Floor When Remodeling Science Labs

    ERIC Educational Resources Information Center

    Nation's Schools, 1972

    1972-01-01

    A new remodeling idea adopts the concept of raised floor covering gas, water, electrical, and drain lines. The accessible floor has removable panels set into an adjustable support frame 24 inches above a concrete subfloor. (Author)

  2. Could Parkinson's Disease Raise Stroke Risk?

    MedlinePlus

    ... news/fullstory_163751.html Could Parkinson's Disease Raise Stroke Risk? Or is the link the other way ... link between Parkinson's disease and the risk for stroke. However, the study can't prove that one ...

  3. Gfi1Cre mice have early onset progressive hearing loss and induce recombination in numerous inner ear non-hair cells

    PubMed Central

    Matern, Maggie; Vijayakumar, Sarath; Margulies, Zachary; Milon, Beatrice; Song, Yang; Elkon, Ran; Zhang, Xiaoyu; Jones, Sherri M.; Hertzano, Ronna

    2017-01-01

    Studies of developmental and functional biology largely rely on conditional expression of genes in a cell type-specific manner. Therefore, the importance of specificity and lack of inherent phenotypes for Cre-driver animals cannot be overemphasized. The Gfi1Cre mouse is commonly used for conditional hair cell-specific gene deletion/reporter gene activation in the inner ear. Here, using immunofluorescence and flow cytometry, we show that the Gfi1Cre mice produce a pattern of recombination that is not strictly limited to hair cells within the inner ear. We observe a broad expression of Cre recombinase in the Gfi1Cre mouse neonatal inner ear, primarily in inner ear resident macrophages, which outnumber the hair cells. We further show that heterozygous Gfi1Cre mice exhibit an early onset progressive hearing loss as compared with their wild-type littermates. Importantly, vestibular function remains intact in heterozygotes up to 10 months, the latest time point tested. Finally, we detect minor, but statistically significant, changes in expression of hair cell-enriched transcripts in the Gfi1Cre heterozygous mice cochleae compared with their wild-type littermate controls. Given the broad use of the Gfi1Cre mice, both for gene deletion and reporter gene activation, these data are significant and necessary for proper planning and interpretation of experiments. PMID:28181545

  4. Osterix-cre labeled progenitor cells contribute to the formation and maintenance of the bone marrow stroma.

    PubMed

    Liu, Yaling; Strecker, Sara; Wang, Liping; Kronenberg, Mark S; Wang, Wen; Rowe, David W; Maye, Peter

    2013-01-01

    We have carried out fate mapping studies using Osterix-EGFPCre and Osterix-CreERt animal models and found Cre reporter expression in many different cell types that make up the bone marrow stroma. Constitutive fate mapping resulted in the labeling of different cellular components located throughout the bone marrow, whereas temporal fate mapping at E14.5 resulted in the labeling of cells within a region of the bone marrow. The identity of cell types marked by constitutive and temporal fate mapping included osteoblasts, adipocytes, vascular smooth muscle, perineural, and stromal cells. Prolonged tracing of embryonic precursors labeled at E14.5dpc revealed the continued existence of their progeny up to 10 months of age, suggesting that fate mapped, labeled embryonic precursors gave rise to long lived bone marrow progenitor cells. To provide further evidence for the marking of bone marrow progenitors, bone marrow cultures derived from Osterix-EGFPCre/Ai9 mice showed that stromal cells retained Cre reporter expression and yielded a FACS sorted population that was able to differentiate into osteoblasts, adipocytes, and chondrocytes in vitro and into osteoblasts, adipocytes, and perivascular stromal cells after transplantation. Collectively, our studies reveal the developmental process by which Osterix-Cre labeled embryonic progenitors give rise to adult bone marrow progenitors which establish and maintain the bone marrow stroma.

  5. Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti

    PubMed Central

    Häcker, Irina; Harrell II, Robert A.; Eichner, Gerrit; Pilitt, Kristina L.; O’Brochta, David A.; Handler, Alfred M.; Schetelig, Marc F.

    2017-01-01

    Site-specific genome modification (SSM) is an important tool for mosquito functional genomics and comparative gene expression studies, which contribute to a better understanding of mosquito biology and are thus a key to finding new strategies to eliminate vector-borne diseases. Moreover, it allows for the creation of advanced transgenic strains for vector control programs. SSM circumvents the drawbacks of transposon-mediated transgenesis, where random transgene integration into the host genome results in insertional mutagenesis and variable position effects. We applied the Cre/lox recombinase-mediated cassette exchange (RMCE) system to Aedes aegypti, the vector of dengue, chikungunya, and Zika viruses. In this context we created four target site lines for RMCE and evaluated their fitness costs. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. aegypti. The advantages of Cre-RMCE over existing site-specific modification systems for Ae. aegypti, phiC31-RMCE and CRISPR, originate in the preservation of the recombination sites, which 1) allows successive modifications and rapid expansion or adaptation of existing systems by repeated targeting of the same site; and 2) provides reversibility, thus allowing the excision of undesired sequences. Thereby, Cre-RMCE complements existing genomic modification tools, adding flexibility and versatility to vector genome targeting. PMID:28266580

  6. Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti.

    PubMed

    Häcker, Irina; Harrell Ii, Robert A; Eichner, Gerrit; Pilitt, Kristina L; O'Brochta, David A; Handler, Alfred M; Schetelig, Marc F

    2017-03-07

    Site-specific genome modification (SSM) is an important tool for mosquito functional genomics and comparative gene expression studies, which contribute to a better understanding of mosquito biology and are thus a key to finding new strategies to eliminate vector-borne diseases. Moreover, it allows for the creation of advanced transgenic strains for vector control programs. SSM circumvents the drawbacks of transposon-mediated transgenesis, where random transgene integration into the host genome results in insertional mutagenesis and variable position effects. We applied the Cre/lox recombinase-mediated cassette exchange (RMCE) system to Aedes aegypti, the vector of dengue, chikungunya, and Zika viruses. In this context we created four target site lines for RMCE and evaluated their fitness costs. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. aegypti. The advantages of Cre-RMCE over existing site-specific modification systems for Ae. aegypti, phiC31-RMCE and CRISPR, originate in the preservation of the recombination sites, which 1) allows successive modifications and rapid expansion or adaptation of existing systems by repeated targeting of the same site; and 2) provides reversibility, thus allowing the excision of undesired sequences. Thereby, Cre-RMCE complements existing genomic modification tools, adding flexibility and versatility to vector genome targeting.

  7. A Novel Bacterial Artificial Chromosome-Transgenic Podoplanin–Cre Mouse Targets Lymphoid Organ Stromal Cells in vivo

    PubMed Central

    Onder, Lucas; Scandella, Elke; Chai, Qian; Firner, Sonja; Mayer, Christian T.; Sparwasser, Tim; Thiel, Volker; Rülicke, Thomas; Ludewig, Burkhard

    2011-01-01

    Stromal cells provide the structural foundation of secondary lymphoid organs (SLOs), and regulate leukocyte access and cell migration within the different compartments of spleen and lymph nodes (LNs). Furthermore, several stromal cell subsets have been implied in shaping of T cell responses through direct presentation of antigen. Despite significant gain of knowledge on the biology of different SLO-resident stromal cell subsets, their molecular and functional characterization has remained incomplete. To address this need, we have generated a bacterial artificial chromosome-transgenic mouse model that utilizes the podoplanin (pdpn) promoter to express the Cre-recombinase exclusively in stromal cells of SLOs. The characterization of the Pdpn–Cre mouse revealed transgene expression in subsets of fibroblastic reticular cells and lymphatic endothelial cells in LNs. Furthermore, the transgene facilitated the identification of a novel splenic perivascular stromal cell subpopulation that forms web-like structures around central arterioles. Assessment of the in vivo antigen expression in the genetically tagged stromal cells in Pdpn–Cre mice revealed activation of both MHC I and II-restricted TCR transgenic T cells. Taken together, stromal pdpn–Cre expression is well-suited to characterize the phenotype and to dissect the function of lymphoid organ stromal cells. PMID:22566840

  8. Fate Tracing of neurogenin2-Expressing Cells in the Mouse Retina Using CreER™: LacZ

    PubMed Central

    Ma, Wenxin; Wang, Shu-Zhen

    2012-01-01

    Delineating the final fate of progenitor cells that transiently express a regulatory gene may shed light on how the gene participates in regulating retinal development. We describe the steps in tracing final fates of progenitor cells that once transiently express neurogenin2 (ngn2) during mouse retinal development with the binary, conditional Ngn2-CreER™—LacZ reporter system. Ngn2-CreER™ mice (Zirlinger et al. Proc Natl Acad Sci USA 99:8084–8089, 2002), in which ngn2 promoter drives the expression of Cre-estrogen receptor CreER™ (Littlewood et al. Nuc Acid Res 23:1686–1690, 1995; Hayashi and McMahon Dev Biol 244:305–318, 2002), are crossed with Rosa26-LoxP-LacZ reporter mice (Soriano Nat Genet 21:70–71, 1999), in which the expression of lacZ requires the removal of “stop” by Cre recombinase (Wagner et al. Transgenic Res 10:545–553, 2001). 4-hydroxytamoxifen (4-OHT), a synthetic ligand with high affinity for ER™, is administered to double transgenic embryos and/or neonatal mice. Binding of 4-OHT to Cre-ER™ activates Cre recombinase, which then catalyzes the removal of the “stop” sequence from the LoxP-LacZ transgene, leading to lacZ expression in cells that express ngn2. Retinal tissues are fixed at different time points after 4-OHT treatment and analyzed for LacZ activities by colorimetric reaction. Double-labeling with a cell type-specific marker can be used to define the identity of a LacZ+ cell. Combining persisted lacZ expression through the life of the cell and the short half-life (0.5–2 h) of 4-OHT (Danielian et al. Curr Biol 8:1323–1326, 1998), this system offers the opportunity to track the final fates of cells that have expressed ngn2 during the brief presence of 4-OHT administered during retinal development. PMID:22688703

  9. Advanced strategic missile development

    NASA Astrophysics Data System (ADS)

    Strickler, R. L.

    1981-05-01

    The M-X program is taking two paths: (1) the current development and projected deployment of a survivable land based ICBM (the M-X) in a multiple protective structure system, and (2) a building block development of readiness posture and strategic futures technology that could be used for a wide range of projected needs in the event of major changes in the threat or the political climate. The blend of aerospace and civil engineering technologies which has resulted in the systems concept necessary to assure the continued survivability of the land based strategic missile force is summarized. Recent advanced technology development activities, which have been focused on systems upgrade options to the current ICBM force, basing options which may be required for special force elements, small missile options for airborne applications, penetration technology to counter SAM and ABM threats, and systems concepts for unique targeting requirements are reviewed.

  10. Information Technology Strategic Plan

    DTIC Science & Technology

    1998-06-01

    The members of the Information Technology Steering Group (ITSG) have been meeting since January 1998 to support the Space and Naval Warfare (SPAWAR...for corporate information technology (IT). The IT Strategic Plan documents the role that corporate information technology plays in achieving SSC San...Diego’s mission, vision and goals. This plan defines a vision for SSC San Diego’s information technology environment that will enhance the quality of

  11. Installation Strategic Planning Guidebook

    DTIC Science & Technology

    2012-05-01

    team members can hear each other and not be distracted by outside noise • Access to bathrooms, snacks , beverages • Note the plenary room can often...workshop. The stakeholders are working hard over the 2.5 days, and easy access to beverages, Installation Strategic Planning Guidebook 50 snacks ...for snacks and beverages. In several cases, Garrisons have also arranged for lunch orders (placed and paid for by participants) to be delivered (with

  12. Microgravity strategic plan, 1988

    NASA Technical Reports Server (NTRS)

    1988-01-01

    The NASA agency-wide microgravity strategic plan is presented, and its research, applications, and commercialization for the 1990's is addressed. The plan presents an analysis of the current situation, identifies critical factors, and defines goals, objectives, and strategies, which are intended to: (1) provide a context for decision making; (2) assure realism in long-range planning and direction for hardware development; and (3) establish a framework for developing a national microgravity research plan.

  13. Strategic Leadership Development Model

    DTIC Science & Technology

    2012-03-19

    meditation .1 —Napoleon Leadership is a subject which excites interest amongst scholars and laymen alike as it is essential for the organized...should also be covered along with intellectual stimulation including the art of objective reflection and meditation . Further subjects suggested for...towards ample leisure time for reflection and meditation . 21  Solicit active participation of corporate representatives in strategic leadership

  14. The French Strategic Dilemma.

    DTIC Science & Technology

    1984-03-01

    crisis . The language of French strategic doctrine is that of dissuasion (deterrence). The French talk publicly much less about warfighting (and employment...threaten vital lines of supply in a crisis situation. Nuclear forces would not *-"e necessarily be useful In deterring Soviet actions in this regard. Rather...34 in Pierre Lellouche, ed., Pacifisme et Dissuasion (Paris: Economica , 1983), pp. 253-266. C. % 6. See Pierre Lacoste, Strategie Navale (Paris: Fernand

  15. A National Strategic Narrative

    DTIC Science & Technology

    2011-01-01

    strategic ecosystem . In other words, the U.S. should stop trying to dominate and direct global events. The best we can do is to build our capital so... China , etc), we need to focus on sustaining ourselves in ways that build our strengths and underpin credible influence. That shift in turn means that...significance of moving toward positive sum politics. To take only one example, the rise of China as a major economic power has been overall very positive

  16. Strategic plan, 1985

    NASA Technical Reports Server (NTRS)

    1985-01-01

    The Lewis Strategic Plan was updated for 1985 and beyond. Major programs for the space station, the advanced turboprop, the Advanced Communications Technology Satellite (ACTS), and the Altitude Wind Tunnel were begun or greatly expanded during 1984. In parallel, The Lewis aeropropulsion research and technology program was extensively evaluated and reviewed; a reduced and reoriented program emerged. The thrusts and implementation plans for these programs are described as they pertain to the individual directorates. Other key accomplishments and plans are summarized.

  17. Making Strategic Analysis Matter

    DTIC Science & Technology

    2012-01-01

    way, so intel- ligence can help them ask, for instance, “Does Plan Colombia offer insights for Afghani- stan?” If the analogy is to be useful, it...Intelligence Agency DNI Director of National Intelligence EADS European Aeronautic Defence and Space Company HIV human immunodeficiency virus NATO...can be a task for strategic analysis. For instance, does Plan Colombia offer any insights by analogy for counterinsurgency in Afghanistan? If

  18. Soviet strategic defense technology

    SciTech Connect

    Stubbs, E.

    1987-04-01

    The present status of the Soviet program suggests several observations that have a bearing on predicting the future of the Soviet strategic defense program and its implications for the US: 1. The Soviet Union appears to have a continuing interest in ABM defenses, although ASATs seem to be a much lower priority. 2. The Soviet technology fielded to date was well within the American grasp 10 years ago. Where advanced and as yet undeployed technologies are concerned, the difference seems to be smaller; perhaps as little as five or seven years, with approximate parity in particle-beam research. 3. The Soviet Union, possibly more sensitive to prestige considerations, appears to be much more inclined than the US to demonstrate and deploy a technology before it is actually fully operational, and to undertake field modifications later. They also are much more reluctant to retire aging and obsolete technologies. As a result, they presently possess the world's only deployed ASAT and ABM systems, however, doubtful their actual operational effectiveness might be. 4. Soviet strategic defenses tend to be more fragmentary in design, reflecting their difficulties with the supporting and integrative technologies such as sensing, signal processing, heavy-lift boosters, and computing hardware and software. 5. The Soviets should also be expected to explore alternative avenues of near-term response to SDI, for example by expanding their strategic nuclear arsenal. 28 references, 1 figure, 2 tables.

  19. CRE dating on the scarps of large landslides affecting the Belledonne massif ( French Alps)

    NASA Astrophysics Data System (ADS)

    Lebrouc, V.; Baillet, L.; Schwartz, S.; Jongmans, D.; Gamond, J. F.; Bourles, D.; Le Roux, O.; Carcaillet, J.; Braucher, R.

    2012-04-01

    The southwestern edge of the Belledonne Massif (French Alps) consists of micaschists unconformably covered with Mesozoic sediments and Quaternary deposits. The morphology corresponds to a glacial plateau (Mont Sec plateau) bordered by steep slopes (around 40°), where moraines and peat bog subsist. The massif is incised by the East-West trending Romanche valley that was shaped by several cycles of quaternary glaciations and deglaciations. Slopes are affected by several active or past large scale rock mass instabilities. Cosmic Ray Exposure (CRE) dating was applied on the head scarps of three large landslides, one of which being the active Séchilienne landslide whose headscarp was already dated by Leroux et al. [2009]. Dating results suggest a concomitant initiation of these instabilities at about 7 ± 2 10Be ka, thousands years after the total downwastage of the valley. A different kinematic behaviour was however observed on two contiguous landslides for which continuous exposure profiles were obtained. On the Séchilienne landslide, 23 samples were collected from internal and lateral scarps, as well as on polished bedrock surfaces, with the aim of dating the internal kinematics of the landslide. Preliminary dating results obtained on polished surfaces and near the top of the scarps show unexpected low 10Be concentrations, suggesting the existence of thin moraine or peat bog deposits masking the bedrock, which have been subsequently eroded. The minimum thickness of these deposits was estimated assuming a constant denudation rate over time. Exposure date profiles show that the studied lateral and internal scarps were initiated at the same period as the Sechilienne headscarp. An increase in the exposure rate was also observed between 2 and 1 ka, in agreement with that evidenced along the headscarp. Forty other samples have been collected in the landslide to corroborate these results. Reference Le Roux, O., S. Schwartz , J.-F. Gamond, D. Jongmans, D. Bourles, R

  20. Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes.

    PubMed

    Koehler, Sybille; Brähler, Sebastian; Braun, Fabian; Hagmann, Henning; Rinschen, Markus M; Späth, Martin R; Höhne, Martin; Wunderlich, F Thomas; Schermer, Bernhard; Benzing, Thomas; Brinkkoetter, Paul T

    2017-02-07

    Podocyte injury is a key event in glomerular disease leading to proteinuria and opening the path toward glomerular scarring. As a consequence, glomerular research strives to discover molecular mechanisms and signaling pathways affecting podocyte health. The hNphs2.Cre mouse model has been a valuable tool to manipulate podocyte-specific genes and to label podocytes for lineage tracing and purification. Here we designed a novel podocyte-specific tricistronic Cre mouse model combining codon improved Cre expression and fluorescent cell labeling with mTomato under the control of the endogenous Nphs2 promoter using viral T2A-peptides. Independent expression of endogenous podocin, codon improved Cre, and mTomato was confirmed by immunofluorescence, fluorescent activated cell sorting and protein analyses. Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) mice developed normally and did not show any signs of glomerular disease or off-target effects under basal conditions and in states of disease. Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type)-mediated gene recombination was superior to conventional hNphs2.Cre mice-mediated gene recombination. Last, we compared Cre efficiency in a disease model by mating Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) and hNphs2.Cre mice to Phb2(fl/fl) mice. The podocyte-specific Phb2 knockout by Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) mice resulted in an aggravated glomerular injury as compared to a podocyte-specific Phb2 gene deletion triggered by hNphs2.Cre. Thus, we generated the first tricistronic podocyte mouse model combining enhanced Cre recombinase efficiency and fluorescent labeling in podocytes without the need for additional matings with conventional reporter mouse lines.

  1. Development of myenteric cholinergic neurons in ChAT-Cre;R26R-YFP mice.

    PubMed

    Hao, Marlene M; Bornstein, Joel C; Young, Heather M

    2013-10-01

    Cholinergic neurons are the major excitatory neurons of the enteric nervous system (ENS), and include intrinsic sensory neurons, interneurons, and excitatory motor neurons. Cholinergic neurons have been detected in the embryonic ENS; however, the development of these neurons has been difficult to study as they are difficult to detect prior to birth using conventional immunohistochemistry. In this study we used ChAT-Cre;R26R-YFP mice to examine the development of cholinergic neurons in the gut of embryonic and postnatal mice. Cholinergic (YFP+) neurons were first detected at embryonic day (E)11.5, and the proportion of cholinergic neurons gradually increased during pre- and postnatal development. At birth, myenteric cholinergic neurons comprised less than half of their adult proportions in the small intestine (25% of myenteric neurons were YFP+ at P0 compared to 62% in adults). The earliest cholinergic neurons appear to mainly project anally. Projections into the presumptive circular muscle were first observed at E14.5. A subpopulation of cholinergic neurons coexpress calbindin through embryonic and postnatal development, but only a small proportion coexpressed neuronal nitric oxide synthase. Our study shows that cholinergic neurons in the ENS develop over a protracted period of time.

  2. Camk2a-Cre-mediated conditional deletion of chromatin remodeler Brg1 causes perinatal hydrocephalus.

    PubMed

    Cao, Mou; Wu, Jiang I

    2015-06-15

    Mammalian SWI/SNF-like BAF chromatin remodeling complexes are essential for many aspects of neural development. Mutations in the genes encoding the core subunit Brg1/SmarcA4 or other complex components cause neurodevelopmental diseases and are associated with autism. Congenital hydrocephalus is a serious brain disorder often experienced by these patients. We report a role of Brg1 in the pathogenesis of hydrocephalus disorder. We discovered an unexpected early activity of mouse Camk2a-Cre transgene, which mediates Brg1 deletion in a subset of forebrain neurons beginning in the late embryonic stage. Brg1 deletion in these neurons led to severe congenital hydrocephalus with enlargement of the lateral ventricles and attenuation of the cerebral cortex. The Brg1-deficient mice had significantly smaller subcommissural organs and narrower Sylvian aqueducts than mice that express normal levels of Brg1. Effects were non-cell autonomous and may be responsible for the development of the congenital hydrocephalus phenotype. Our study provides evidence indicating that abnormalities in Brg1 function result in defects associated with neurodevelopmental disorders and autism.

  3. Cyclic AMP induces IPC leukemia cell apoptosis via CRE-and CDK-dependent Bim transcription.

    PubMed

    Huseby, S; Gausdal, G; Keen, T J; Kjærland, E; Krakstad, C; Myhren, L; Brønstad, K; Kunick, C; Schwede, F; Genieser, H-G; Kleppe, R; Døskeland, S O

    2011-12-08

    The IPC-81 cell line is derived from the transplantable BNML model of acute myelogenic leukemia (AML), known to be a reliable predictor of the clinical efficiency of antileukemic agents, like the first-line AML anthracycline drug daunorubicin (DNR). We show here that cAMP acted synergistically with DNR to induce IPC cell death. The DNR-induced death differed from that induced by cAMP by (1) not involving Bim induction, (2) being abrogated by GSK3β inhibitors, (3) by being promoted by the HSP90/p23 antagonist geldanamycin and truncated p23 and (4) by being insensitive to the CRE binding protein (CREB) antagonist ICER and to cyclin-dependent protein kinase (CDK) inhibitors. In contrast, the apoptosis induced by cAMP correlated tightly with Bim protein expression. It was abrogated by Bim (BCL2L11) downregulation, whether achieved by the CREB antagonist ICER, by CDK inhibitors, by Bim-directed RNAi, or by protein synthesis inhibitor. The forced expression of BimL killed IPC-81(WT) cells rapidly, Bcl2-overexpressing cells being partially resistant. The pivotal role of CREB and CDK activity for Bim transcription is unprecedented. It is also noteworthy that newly developed cAMP analogs specifically activating PKA isozyme I (PKA-I) were able to induce IPC cell apoptosis. Our findings support the notion that AML cells may possess targetable death pathways not exploited by common anti-cancer agents.

  4. Cylindrically symmetric wormholes WhCRe : The motion of test particles

    NASA Astrophysics Data System (ADS)

    Aminova, Asya V.; Brill, Dieter R.; Chumarov, Pavel I.; Shemakhin, Aleksandr Yu.

    2016-07-01

    We study the radial and nonradial motion of massive test particles and photons in a three-parameter family of cylindrically symmetric wormholes WhCRe generated by the electromagnetic, dilaton, and scalar fields, with particular attention paid to the extent to which the wormhole is traversable. The wormholes are not asymptotically flat and contain a curvature singularity at one end of the wormhole. In the case of nonradial motion with conserved energy and angular momentum along a hypersurface z =const ("planar orbits") we show that, as in the Kerr and Schwarzschild geometries, we should distinguish between orbits with impact parameters greater or less than a certain critical value Dc, which corresponds to the unstable circular orbit of radius uc. For D2>Dc2 there are two kinds of orbits: orbits of the first kind arrive from infinity and turn around at the orbit's minimum radial coordinate u ("pericenter") greater than uc, whereas orbits of the second kind turn around at maximum radial coordinate u ("apocenter") less than uc and terminate at the singularity at u =-∞ . For D =Dc orbits of the first and second kinds merge and both orbits spiral an infinite number of times toward the unstable circular orbit u =uc. For D2

  5. Expression of I-CreI Endonuclease Generates Deletions Within the rDNA of Drosophila

    PubMed Central

    Paredes, Silvana; Maggert, Keith A.

    2009-01-01

    The rDNA arrays in Drosophila contain the cis-acting nucleolus organizer regions responsible for forming the nucleolus and the genes for the 28S, 18S, and 5.8S/2S RNA components of the ribosomes and so serve a central role in protein synthesis. Mutations or alterations that affect the nucleolus organizer region have pleiotropic effects on genome regulation and development and may play a role in genomewide phenomena such as aging and cancer. We demonstrate a method to create an allelic series of graded deletions in the Drosophila Y-linked rDNA of otherwise isogenic chromosomes, quantify the size of the deletions using real-time PCR, and monitor magnification of the rDNA arrays as their functions are restored. We use this series to define the thresholds of Y-linked rDNA required for sufficient protein translation, as well as establish the rate of Y-linked rDNA magnification in Drosophila. Finally, we show that I-CreI expression can revert rDNA deletion phenotypes, suggesting that double-strand breaks are sufficient to induce rDNA magnification. PMID:19171942

  6. Does strategic defense breed offense

    SciTech Connect

    York, H.

    1987-01-01

    The author examines the question of whether strategic defense activity, in any form, stimulates a related offensive activity. The author studies four post-World II efforts of the superpowers to develop and deploy strategic defenses. The author then derives lessons from those cases that he applies to his analysis of President Reagan's Strategic Defense Initiative. Commentaries on the author's analysis are provided by four scholars.

  7. Strategic Military Leaders - Leading Tomorrow

    DTIC Science & Technology

    2008-02-29

    strategic vision, and prepare their commands as a whole for their future roles .”7 Transiting from an environment of relative clarity in missions...in war, their role is to dominate and win. Social Intelligence Daniel Goleman defines social intelligence as a combination of two inseparable...therefore has two crucial roles : first to develop the right people, with the right strategic competencies, as its strategic leaders; and second, to

  8. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    Not Available

    1990-08-15

    The Strategic Petroleum Reserve Quarterly Report is submitted in accordance with section 165(b) of the Energy Policy and Conservation Act, as amended, which requires that the Secretary of Energy submit quarterly reports to Congress on Activities undertaken with respect to the Strategic Petroleum Reserve. This August 15, 1990, Strategic Petroleum Reserve Quarterly Report describes activities related to the site development, oil acquisition, budget and cost of the Reserve during the period April 1, 1990, through June 30, 1990. 3 tabs.

  9. Dermal Delivery of Constructs Encoding Cre Recombinase to Induce Skin Tumors in PtenLoxP/LoxP;BrafCA/+ Mice

    PubMed Central

    Deken, Marcel A.; Song, Ji-Ying; Gadiot, Jules; Bins, Adriaan D.; Kroon, Paula; Verbrugge, Inge; Blank, Christian U.

    2016-01-01

    Current genetically-engineered mouse melanoma models are often based on Tyr::CreERT2-controlled MAPK pathway activation by the BRAFV600E mutation and PI3K pathway activation by loss of PTEN. The major drawback of these models is the occurrence of spontaneous tumors caused by leakiness of the Tyr::CreERT2 system, hampering long-term experiments. To address this problem, we investigated several approaches to optimally provide local delivery of Cre recombinase, including injection of lentiviral particles, DNA tattoo administration and particle-mediated gene transfer, to induce melanomas in PtenLoxP/LoxP;BrafCA/+ mice lacking the Tyr::CreERT2 allele. We found that dermal delivery of the Cre recombinase gene under the control of a non-specific CAG promoter induced the formation of melanomas, but also keratoacanthoma and squamous cell carcinomas. Delivery of Cre recombinase DNA under the control of melanocyte-specific promoters in PtenLoxP/LoxP;BrafCA/+ mice resulted in sole melanoma induction. The growth rate and histological features of the induced tumors were similar to 4-hydroxytamoxifen-induced tumors in Tyr::CreERT2;PtenLoxP/LoxP;BrafCA/+ mice, while the onset of spontaneous tumors was prevented completely. These novel induction methods will allow long-term experiments in mouse models of skin malignancies. PMID:27999416

  10. The CRE1 Cytokinin Pathway Is Differentially Recruited Depending on Medicago truncatula Root Environments and Negatively Regulates Resistance to a Pathogen

    PubMed Central

    Laffont, Carole; Rey, Thomas; André, Olivier; Novero, Mara; Kazmierczak, Théophile; Debellé, Frédéric; Bonfante, Paola; Jacquet, Christophe; Frugier, Florian

    2015-01-01

    Cytokinins are phytohormones that regulate many developmental and environmental responses. The Medicago truncatula cytokinin receptor MtCRE1 (Cytokinin Response 1) is required for the nitrogen-fixing symbiosis with rhizobia. As several cytokinin signaling genes are modulated in roots depending on different biotic and abiotic conditions, we assessed potential involvement of this pathway in various root environmental responses. Phenotyping of cre1 mutant roots infected by the Gigaspora margarita arbuscular mycorrhizal (AM) symbiotic fungus, the Aphanomyces euteiches root oomycete, or subjected to an abiotic stress (salt), were carried out. Detailed histological analysis and quantification of cre1 mycorrhized roots did not reveal any detrimental phenotype, suggesting that MtCRE1 does not belong to the ancestral common symbiotic pathway shared by rhizobial and AM symbioses. cre1 mutants formed an increased number of emerged lateral roots compared to wild-type plants, a phenotype which was also observed under non-stressed conditions. In response to A. euteiches, cre1 mutants showed reduced disease symptoms and an increased plant survival rate, correlated to an enhanced formation of lateral roots, a feature previously linked to Aphanomyces resistance. Overall, we showed that the cytokinin CRE1 pathway is not only required for symbiotic nodule organogenesis but also affects both root development and resistance to abiotic and biotic environmental stresses. PMID:25562779

  11. Winning Techniques for Athletic Fund Raising.

    ERIC Educational Resources Information Center

    Alberger, Patti, Ed.

    Techniques are presented from successful athletic fund-raisers on how to structure a fund-raising program through an independent booster club, as part of the athletic department or as part of the total institutional development program. The handbook offers advice on building a program from scratch; organizing successful campaigns, involving…

  12. Raising Confident, Competent Daughters: Strategies for Parents.

    ERIC Educational Resources Information Center

    Ransome, Whitney, Ed.; And Others

    This booklet contains five essays designed to help parents raise confident, competent daughters. They focus on ways that parents can help their preadolescent and adolescent daughters: (1) speak up in class, articulate their thoughts, and speak with self-confidence in various academic and social situations; (2) develop an interest and aptitude for…

  13. Raising Butterflies from Your Own Garden.

    ERIC Educational Resources Information Center

    Howley-Pfeifer, Patricia

    2002-01-01

    Describes how raising monarch, black swallowtail, and mourning cloak butterflies in a kindergarten class garden can provide opportunities for observation experiences. Includes detailed steps for instruction and describes stages of growth. Excerpts children's journal dictations to illustrate ways to support the discovery process. Describes related…

  14. Guidelines Manual for Fund Raising and Donations.

    ERIC Educational Resources Information Center

    Tidewater Community Coll. Educational Foundation, Inc., Portsmouth, VA.

    Guidelines and procedures for fund raising and the acceptance of gifts at Tidewater Community College (TCC) are presented in this manual. First, general information is provided on the acceptance of gifts of cash, securities, real estate, insurance, personal property, equipment, and gifts requiring the expenditure of funds. Next, general procedures…

  15. Consciousness-Raising, Error Correction and Proofreading

    ERIC Educational Resources Information Center

    O'Brien, Josephine

    2015-01-01

    The paper discusses the impact of developing a consciousness-raising approach in error correction at the sentence level to improve students' proofreading ability. Learners of English in a foreign language environment often rely on translation as a composing tool and while this may act as a scaffold and provide some support, it frequently leads to…

  16. Raising Children Bilingually Is Hard: Why Bother?

    ERIC Educational Resources Information Center

    Sims, Margaret; Ellis, Elizabeth M.

    2014-01-01

    This paper presents results from a pilot project that sought to establish parental motivations for raising children bilingually in regional Australia in the absence of a co-located speech community. Cultural and linguistic diversity outside metropolitan areas is increasing as a result of Commonwealth Government incentive schemes, and one effect of…

  17. Some Blunt Talk about Fund Raising.

    ERIC Educational Resources Information Center

    Buchanan, Peter McE.

    1991-01-01

    The president of the Council for the Advancement and Support of Education talks about why college advancement officers need to rethink, restructure, and communicate better about their fund-raising campaigns, in light of public perceptions that the institutions are greedy and not managing resources efficiently. Emphasis is on carefully defining…

  18. YOU CAN RAISE YOUR HANDICAPPED CHILD.

    ERIC Educational Resources Information Center

    AYRAULT, EVELYN WEST

    DESIGNED AS A GUIDE FOR PARENTS OF HANDICAPPED CHILDREN, THIS BOOK PROVIDES INFORMATION ON TYPES OF HANDICAPS AND THE AIMS AND TECHNIQUES OF TREATMENT, THERAPY, AND EDUCATION. SPECIFIC RECOMMENDATIONS TO HELP PARENTS RAISE A HANDICAPPED CHILD ARE DISCUSSED. BEHAVIOR PROBLEMS, PARENT ATTITUDES, DAILY PROBLEMS, AND RECREATION ARE EXAMINED IN DETAIL…

  19. People on the Farm: Raising Beef Cattle.

    ERIC Educational Resources Information Center

    Crain, Robert L.

    This booklet provides information on raising beef cattle through profiles of two families, the Ritschards of Colorado and the Schuttes of Missouri. Through descriptions of daily life for these families, the booklet discusses the way of life on modern beef cattle farms and the problems and decisions faced by farmers. The booklet explains how…

  20. Raising Environmental Awareness among Miners in Iran

    ERIC Educational Resources Information Center

    Mozaffari, Ezatollah

    2013-01-01

    Generation of waste is inevitable but controllable in minerals industry. The aim of this research is to find ways for raising environmental awareness among miners. Miners' attitude towards environmental mining has been investigated. A survey has been done collecting mine managers' point of view coupled with current trend on mine waste management…

  1. The Power of the Raised Eyebrow

    ERIC Educational Resources Information Center

    Burton, Grace M.

    1977-01-01

    The school counselor is in a unique position to help provide equal education for all students particularly when it comes to high school girls choosing high level mathematics courses. The counselor should encourage young women to take courses in mathematics and the sciences throughout their high school careers and never, never raise an eyebrow!…

  2. Liberation Therapeutics: Consciousness Raising as a Problem.

    ERIC Educational Resources Information Center

    Lasch-Quinn, Elisabeth

    2002-01-01

    Questions the content and form of consciousness raising as a mode of purveying knowledge or bringing about change by considering its emergence in the civil rights movement. Examines such books as "Black Rage" (William Grierand Price Cobbs), "Triumph of the Therapeutic" (Philip Reiff), "Where Do We Go From Here: Chaos or…

  3. The Fundamentals of Community College Fund Raising.

    ERIC Educational Resources Information Center

    Brumbach, Mary A.; Bumphus, Walter G.

    1993-01-01

    Presents a guide for community college fund raising. Discusses organizational principles and structural models; operational concerns such as stewardship of funds, information dissemination, and donor research; and the importance of presidential support, institutional reputation and environment, and experienced grants managers. Includes practical…

  4. Pragmatics, Awareness Raising, and the Cooperative Principle

    ERIC Educational Resources Information Center

    Murray, Neil

    2010-01-01

    In recent years, pedagogical pragmatics has sought to improve the effectiveness with which learners express and interpret meaning, through awareness-raising activities that draw on authentic materials and break away from simplistic explanations of form-function correspondences. By and large, these efforts have been informed by an inductive…

  5. National Organizations Raising Public Consciousness. Part II.

    ERIC Educational Resources Information Center

    Exceptional Parent, 1980

    1980-01-01

    Statements of seven national organizations (Muscular Dystrophy Association, Spina Bifida Association of America, National Easter Seal Society, International Association of Parents of the Deaf, American Foundation for the Blind, Down's Syndrome Congress, and the National Society for Autistic Children) about raising community awareness of issues…

  6. Raising Money Through an Institutionally Related Foundation.

    ERIC Educational Resources Information Center

    Reilley, Timothy A., Ed.

    The creation of foundations for fund raising at public colleges and new ideas and techniques for established foundations are discussed in 13 chapters. The relationship of the foundation and the institution is described from the viewpoint of the institution and also that of the foundation. Article titles and authors include: "How the…

  7. Raising Happy Kids on a Reasonable Budget.

    ERIC Educational Resources Information Center

    Gallagher, Patricia C.

    This guide shows parents how to cut expenses and save on the cost of raising a family. Chapter 1 offers many ways to stretch food dollars and discusses buying in bulk, eating out, using coupons, buying wholesale, and gardening. Cost-cutting clothing ideas covered in chapter 2 include buying at secondhand stores and consignment shops, outlet…

  8. Solo Parenting: Raising Strong & Happy Families.

    ERIC Educational Resources Information Center

    Chambers, Diane

    One quarter of all American children will live in a single-parent family for at least some portion of their childhood. This guide, organized as a step-by-step process, encourages single parents to reach for excellence in their parenting and to build confidence in their ability to raise healthy, responsible children. The chapters are intended to…

  9. Issues Raised by the Follow Through Evaluation.

    ERIC Educational Resources Information Center

    House, Ernest R.; Hutchins, Elizabeth J.

    This paper presents a discussion of issues raised in the evaluation of Project Follow Through reported by Abt Associates. The paper suggests that many of the problems inherent in the design of both the program and the evaluation stem from the underlying assumption that one educational model could be found which would best alleviate the educational…

  10. Direct involvement of the CreA transcription factor in penicillin biosynthesis and expression of the pcbAB gene in Penicillium chrysogenum.

    PubMed

    Cepeda-García, Cristina; Domínguez-Santos, Rebeca; García-Rico, Ramón O; García-Estrada, Carlos; Cajiao, Angela; Fierro, Francisco; Martín, Juan Francisco

    2014-08-01

    The transcription factor CreA is the main regulator responsible for carbon repression in filamentous fungi. CreA is a wide domain regulator that binds to regulatory elements in the promoters of target genes to repress their transcription. Penicillin biosynthesis and the expression of penicillin biosynthetic genes are subject to carbon repression. However, evidence of the participation of CreA in this regulation is still lacking, and previous studies on the promoter of the pcbC gene of Aspergillus nidulans indicated the lack of involvement of CreA in its regulation. Here we present clear evidence of the participation of CreA in carbon repression of penicillin biosynthesis and expression of the pcbAB gene, encoding the first enzyme of the pathway, in Penicillium chrysogenum. Mutations in cis of some of the putative CreA binding sites present in the pcbAB gene promoter fused to a reporter gene caused an important increase in the measured enzyme activity in glucose-containing medium, whereas activity in the medium with lactose was not affected. An RNAi strategy was used to attenuate the expression of the creA gene. Transformants expressing a small interfering RNA for creA showed higher penicillin production, and this increase was more evident when glucose was used as carbon source. These results confirm that CreA plays an important role in the regulation of penicillin biosynthesis in P. chrysogenum and opens the possibility of its utilization to improve the industrial production of this antibiotic.

  11. 75 FR 67695 - U.S. Strategic Command Strategic Advisory Group Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-03

    ... to the Commander, U.S. Strategic Command, during the development of the Nation's strategic war plans... of the Secretary of Defense U.S. Strategic Command Strategic Advisory Group Closed Meeting AGENCY.... Strategic Command Strategic Advisory Group. DATES: December 9, 2010: 8 a.m. to 5 p.m. December 10, 2010: 8...

  12. Children Seem to Know Raising: Raising and Intervention in Child Language

    ERIC Educational Resources Information Center

    Choe, Jinsun

    2012-01-01

    English-speaking children exhibit difficulty in their comprehension of raising patterns, such as (1), in which the NP the boy is semantically linked to the VP in the embedded clause, but is syntactically realized as the subject of the matrix clause. (1) Raising pattern: [s "The boy" seems to the girl [s _ to be happy

  13. Strategic implementation plan

    NASA Technical Reports Server (NTRS)

    1989-01-01

    The Life Science Division of the NASA Office of Space Science and Applications (OSSA) describes its plans for assuring the health, safety, and productivity of astronauts in space, and its plans for acquiring further fundamental scientific knowledge concerning space life sciences. This strategic implementation plan details OSSA's goals, objectives, and planned initiatives. The following areas of interest are identified: operational medicine; biomedical research; space biology; exobiology; biospheric research; controlled ecological life support; flight programs and advance technology development; the life sciences educational program; and earth benefits from space life sciences.

  14. NASA strategic plan

    NASA Technical Reports Server (NTRS)

    1995-01-01

    NASA's Plan summarizes the Agency's vision, mission, and values. Specific goals are listed for each externally focused Enterprise: Mission to Planet Earth, Aeronautics, Human Exploration and Development of Space, Space Science, and Space Technology. These Enterprises satisfy the needs of customers external to NASA. The Strategic Functions (Space Communications, Human Resources, and Physical Resources) are necessary in order to meet the goals of the Enterprises. The goals of these Functions are also presented. All goals must be met while adhering to the discussed values and operating principles of NASA. A final section outlines the implementing strategy.

  15. Guam Strategic Energy Plan

    SciTech Connect

    Conrad, M. D.

    2013-07-01

    Describes various energy strategies available to Guam to meet the territory's goal of diversifying fuel sources and reducing fossil energy consumption 20% by 2020.The information presented in this strategic energy plan will be used by the Guam Energy Task Force to develop an energy action plan. Available energy strategies include policy changes, education and outreach, reducing energy consumption at federal facilities, and expanding the use of a range of energy technologies, including buildings energy efficiency and conservation, renewable electricity production, and alternative transportation. The strategies are categorized based on the time required to implement them.

  16. Enhancing Cellulase Production in Thermophilic Fungus Myceliophthora thermophila ATCC42464 by RNA Interference of cre1 Gene Expression.

    PubMed

    Yang, Fan; Gong, Yanfen; Liu, Gang; Zhao, Shengming; Wang, Juan

    2015-07-01

    The role of CRE1 in a thermophilic fungus, Myceliophthora thermophila ATCC42464, was studied using RNA interference. In the cre1-silenced strain C88, the filter paper hydrolyzing activity and β-1,4-endoglucanase activity were 3.76-, and 1.31-fold higher, respectively, than those in the parental strain when the strains were cultured in inducing medium for 6 days. The activities of β-1,4-exoglucanase and cellobiase were 2.64-, and 5.59-fold higher, respectively, than those in the parental strain when the strains were cultured for 5 days. Quantitative reverse-transcription polymerase chain reaction showed that the gene expression of egl3, cbh1, and cbh2 was significantly increased in transformant C88 compared with the wild-type strain. Therefore, our findings suggest the feasibility of improving cellulase production by modifying the regulator expression, and an attractive approach to increasing the total cellulase productivity in thermophilic fungi.

  17. A Self-deleting Cre-lox-ermAM Cassette, CHESHIRE, for marker-less gene deletion in Streptococcus pneumoniae

    PubMed Central

    Weng, Liming; Biswas, Indranil; Morrison, Donald A.

    2009-01-01

    Although targeted mutagenesis of Streptococcus pneumoniae is readily accomplished with the aid of natural genetic transformation and chimeric donor DNA constructs assembled in vitro, the drug resistance markers often employed for selection of recombinant products can themselves be undesirable by-products of the genetic manipulation. A new cassette carrying the erythromycin-resistance marker ermAM is described that can be used as a temporary marker for selection of desired recombinants. The cassette may subsequently be removed at will by virtue of an embedded fucose-regulated Cre recombinase gene and terminal lox66 and lox71 Cre recognition sites, with retention of 34 bp from the cassette as an inert residual double-mutant lox72 site. PMID:19850089

  18. Strategic Posture Annex to the USAF Strategic Master Plan

    DTIC Science & Technology

    2015-05-01

    subject to a penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. 1. REPORT DATE...MAY 2015 2. REPORT TYPE 3. DATES COVERED 00-00-2015 to 00-00-2015 4. TITLE AND SUBTITLE Strategic Posture Annex to the USAF Strategic Master

  19. Strategic Groups: One Perspective on Integrating Strategic and Group Therapies.

    ERIC Educational Resources Information Center

    Leahey, Maureen; Wallace, Evie

    1988-01-01

    Describes a short-term, strategic, solution-focused self-esteem group in an outpatient mental health setting. Discusses constructs that form foundations of both strategic and group therapies. Training, supervision, and outcome issues arising from integrating the two therapies are addressed. (Author/NB)

  20. Strategic Decisions in Task-Oriented Reading.

    PubMed

    Salmerón, Ladislao; Vidal-Abarca, Eduardo; Martínez, Tomás; Mañá, Amelia; Gil, Laura; Naumann, Johannes

    2015-12-28

    Answering questions from texts are assessment and instructional activities that are frequently used in schools. Nevertheless, little is known about the strategic processes that students take while performing these tasks. We explored the amount and frequency that students initially read of a text before they answered questions pertaining to the material. In a procedure similar to the one used in the PISA (Program for International Students Assessment), one-hundred-seventy students between 7th and 9th grade read and answered several questions designed to assess task-oriented reading in three specific texts. We recorded on-line indexes that evaluated student behavior (e.g., the amount of text that students read before answering questions raised within a given text), performance, and comprehension skill. The results revealed that students skilled in comprehension initially read a high proportion of the texts, which in turn improved their overall performance in two of the three texts read (text 1: CI95%: 0.01 to 0.09; text 2: CI95%: -0.01 to 0.05; text 3: CI95%: 0.04 to 0.20). Therefore, we conclude that this strategic behavior should be considered during the assessment and instruction of reading literacy.

  1. Cre recombinase-regulated Endothelin1 transgenic mouse lines: novel tools for analysis of embryonic and adult disorders

    PubMed Central

    Tavares, Andre L.P.; Clouthier, David E.

    2015-01-01

    Endothelin-1 (EDN1) influences both craniofacial and cardiovascular development and a number of adult physiological conditions by binding to one or both of the known endothelin receptors, thus initiating multiple signaling cascades. Animal models containing both conventional and conditional loss of the Edn1 gene have been used to dissect EDN1 function in both embryos and adults. However, while transgenic Edn1 over-expression or targeted genomic insertion of Edn1 has been performed to understand how elevated levels of Edn1 result in or exacerbate disease states, an animal model in which Edn1 over-expression can be achieved in a spatiotemporal-specific manner has not been reported. Here we describe the creation of Edn1 conditional over-expression transgenic mouse lines in which the chicken β-actin promoter and an Edn1 cDNA are separated by a strong stop sequence flanked by loxP sites. In the presence of Cre, the stop cassette is removed, leading to Edn1 expression. Using the Wnt1-Cre strain, in which Cre expression is targeted to the Wnt1-expressing domain of the central nervous system (CNS) from which neural crest cells (NCCs) arise, we show that stable CBA-Edn1 transgenic lines with varying EDN1 protein levels develop defects in NCC-derived tissues of the face, though the severity differs between lines. We also show that Edn1 expression can be achieved in other embryonic tissues utilizing other Cre strains, with this expression also resulting in developmental defects. CBA-Edn1 transgenic mice will be useful in investigating diverse aspects of EDN1-mediated-development and disease, including understanding how NCCs achieve and maintain a positional and functional identity and how aberrant EDN1 levels can lead to multiple physiological changes and diseases. PMID:25725491

  2. SDI (Strategic Defense Initiative): Myth or reality

    SciTech Connect

    Canavan, G.H.

    1988-06-01

    This report reviews previous attempts to develop strategic defenses, the technologies currently under consideration by the Strategic Defense Initiative (SDI), their main unknowns, and the likely performance of strategic defense concepts against evolving threats. 47 refs.

  3. The chaperone-like activity of the hepatitis C virus IRES and CRE elements regulates genome dimerization.

    PubMed

    Romero-López, Cristina; Barroso-delJesus, Alicia; Berzal-Herranz, Alfredo

    2017-02-24

    The RNA genome of the hepatitis C virus (HCV) establishes a network of long-distance RNA-RNA interactions that direct the progression of the infective cycle. This work shows that the dimerization of the viral genome, which is initiated at the dimer linkage sequence (DLS) within the 3'UTR, is promoted by the CRE region, while the IRES is a negative regulatory partner. Using differential 2'-acylation probing (SHAPE-dif) and molecular interference (HMX) technologies, the CRE activity was found to mainly lie in the critical 5BSL3.2 domain, while the IRES-mediated effect is dependent upon conserved residues within the essential structural elements JIIIabc, JIIIef and PK2. These findings support the idea that, along with the DLS motif, the IRES and CRE are needed to control HCV genome dimerization. They also provide evidences of a novel function for these elements as chaperone-like partners that fine-tune the architecture of distant RNA domains within the HCV genome.

  4. The aging behavior of types 308 and 308CRE stainless steels and its effect on mechanical properties

    SciTech Connect

    Vitek, J.M.; David, S.A.

    1987-01-01

    Aging of 308 and 308CRE SS was studied at 475 to 850/sup 0/C for aging times up to 10,000 hours. Above 550/sup 0/C, aging of 308 steel resulted in precipitation of carbides and the transformation of ferrite to sigma phase or the formation of sigma phase in initially ferrite-free material. The elevated-temperature aging of 308CRE steel resulted in the precipitation of titanium-rich carbides, nitrides, and sulfides, and the transformation of ferrite to sigma phase. The distribution of precipitates was affected by the initial condition of the materials. The elevated-temperature creep properties, and in particular the improved properties of 308CRE, were related to the precipitate distribution. Below 550/sup 0/C, aging of welded type 308 steel, precipitation of G-phase within the ferrite was observed, as well as the decomposition of ferrite into alpha and alpha prime. With the help of a novel mechanical properties microprobe, which was capable of determining the hardness of the minor constituent ferrite phase, the hardness behavior as a function of aging could be related to the microstructures. These results are interpreted in terms of the potential susceptibility of these alloys to 475/sup 0/C embrittlement.

  5. The Medicago truncatula CRE1 cytokinin receptor regulates lateral root development and early symbiotic interaction with Sinorhizobium meliloti.

    PubMed

    Gonzalez-Rizzo, Silvina; Crespi, Martin; Frugier, Florian

    2006-10-01

    Legumes develop different types of lateral organs from their primary root, lateral roots and nodules, the latter depending on a symbiotic interaction with Sinorhizobium meliloti. Phytohormones have been shown to function in the control of these organogeneses. However, related signaling pathways have not been identified in legumes. We cloned and characterized the expression of Medicago truncatula genes encoding members of cytokinin signaling pathways. RNA interference of the cytokinin receptor homolog Cytokinin Response1 (Mt CRE1) led to cytokinin-insensitive roots, which showed an increased number of lateral roots and a strong reduction in nodulation. Both the progression of S. meliloti infection and nodule primordia formation were affected. We also identified two cytokinin signaling response regulator genes, Mt RR1 and Mt RR4, which are induced early during the symbiotic interaction. Induction of these genes by S. meliloti infection is altered in mutants affected in the Nod factor signaling pathway; conversely, cytokinin regulation of the early nodulin Nodule Inception1 (Mt NIN) depends on Mt CRE1. Hence, cytokinin signaling mediated by a single receptor, Mt CRE1, leads to an opposite control of symbiotic nodule and lateral root organogenesis. Mt NIN, Mt RR1, and Mt RR4 define a common pathway activated during early S. meliloti interaction, allowing crosstalk between plant cytokinins and bacterial Nod factors signals.

  6. The chaperone-like activity of the hepatitis C virus IRES and CRE elements regulates genome dimerization

    PubMed Central

    Romero-López, Cristina; Barroso-delJesus, Alicia; Berzal-Herranz, Alfredo

    2017-01-01

    The RNA genome of the hepatitis C virus (HCV) establishes a network of long-distance RNA-RNA interactions that direct the progression of the infective cycle. This work shows that the dimerization of the viral genome, which is initiated at the dimer linkage sequence (DLS) within the 3′UTR, is promoted by the CRE region, while the IRES is a negative regulatory partner. Using differential 2′-acylation probing (SHAPE-dif) and molecular interference (HMX) technologies, the CRE activity was found to mainly lie in the critical 5BSL3.2 domain, while the IRES-mediated effect is dependent upon conserved residues within the essential structural elements JIIIabc, JIIIef and PK2. These findings support the idea that, along with the DLS motif, the IRES and CRE are needed to control HCV genome dimerization. They also provide evidences of a novel function for these elements as chaperone-like partners that fine-tune the architecture of distant RNA domains within the HCV genome. PMID:28233845

  7. Evidence from cosmic ray exposure (CRE) dating for the existence of a pre-Minoan caldera on Santorini, Greece

    NASA Astrophysics Data System (ADS)

    Athanassas, C. D.; Bourlès, D. L.; Braucher, R.; Druitt, T. H.; Nomikou, P.; Léanni, L.

    2016-05-01

    Cosmic ray exposure (CRE) dating was performed on the caldera cliffs of Santorini with the aim of detecting cliff segments predating the Minoan eruption (17th century BCE). The methodology involved the determination of in situ-produced cosmogenic 36Cl concentration in basaltic-to-rhyodacitic whole rocks cropping out in the cliffs. After the samples were processed following the chemical protocol of 36Cl preparation for silicate rocks, 36Cl concentrations were measured by accelerator mass spectrometry (AMS). Important challenges during the implementation procedure were related to large amounts of radiogenic 36Cl, complex modeling of inherited 36Cl, and dominance of the thermal and epithermal (low-energy) neutron capture production pathway. Nevertheless, quantitative assessments on the basis of the contribution of the low-energy neutron capture pathway percent to the total production rate validated the calculated CRE dates. Current CRE ages demonstrate that an ancient caldera existed on pre-Minoan Santorini, occupying at least the northern half of the modern-day caldera.

  8. Using the Textpresso Site-Specific Recombinases Web server to identify Cre expressing mouse strains and floxed alleles.

    PubMed

    Condie, Brian G; Urbanski, William M

    2014-01-01

    Effective tools for searching the biomedical literature are essential for identifying reagents or mouse strains as well as for effective experimental design and informed interpretation of experimental results. We have built the Textpresso Site Specific Recombinases (Textpresso SSR) Web server to enable researchers who use mice to perform in-depth searches of a rapidly growing and complex part of the mouse literature. Our Textpresso Web server provides an interface for searching the full text of most of the peer-reviewed publications that report the characterization or use of mouse strains that express Cre or Flp recombinase. The database also contains most of the publications that describe the characterization or analysis of strains carrying conditional alleles or transgenes that can be inactivated or activated by site-specific recombinases such as Cre or Flp. Textpresso SSR complements the existing online databases that catalog Cre and Flp expression patterns by providing a unique online interface for the in-depth text mining of the site specific recombinase literature.

  9. Operationalizing strategic marketing.

    PubMed

    Chambers, S B

    1989-05-01

    The strategic marketing process, like any administrative practice, is far simpler to conceptualize than operationalize within an organization. It is for this reason that this chapter focused on providing practical techniques and strategies for implementing the strategic marketing process. First and foremost, the marketing effort needs to be marketed to the various publics of the organization. This chapter advocated the need to organize the marketing analysis into organizational, competitive, and market phases, and it provided examples of possible designs of the phases. The importance and techniques for exhausting secondary data sources and conducting efficient primary data collection methods were explained and illustrated. Strategies for determining marketing opportunities and threats, as well as segmenting markets, were detailed. The chapter provided techniques for developing marketing strategies, including considering the five patterns of coverage available; determining competitor's position and the marketing mix; examining the stage of the product life cycle; and employing a consumer decision model. The importance of developing explicit objectives, goals, and detailed action plans was emphasized. Finally, helpful hints for operationalizing the communication variable and evaluating marketing programs were provided.

  10. Securitization of Russian Strategic Communication

    DTIC Science & Technology

    2010-12-02

    102 i SCHOOL OF ADVANCED MILITARY STUDIES MONOGRAPH APPROVAL Rank and Student Full Name Title of Monograph: The Securitization of Russian... culture of like-mindedness and the securitization of Russian strategic communications. The increasing securitization of strategic communications brings...state owned corporations at rates that bordered on gifts rather than fair monetary exchange. 15 As Russia underwent this economic struggle, the

  11. Strategic Planning and Information Systems.

    ERIC Educational Resources Information Center

    Shuman, Jack N.

    1982-01-01

    Discusses the functions of business planning systems and analyzes the underlying assumptions of the information systems that support strategic planning efforts within organizations. Development of a system framework, obstacles to the successful creation of strategic planning information systems, and resource allocation in organizations are…

  12. NASA Space Sciences Strategic Planning

    NASA Technical Reports Server (NTRS)

    Crane, Philippe

    2004-01-01

    The purpose of strategic planning roadmap is to:Fulfill the strategic planning requirements; Provide a guide to the science community in presenting research requests to NASA; Inform and inspire; Focus investments in technology and research for future missions; and Provide the scientific and technical justification for augmentation requests.

  13. Strategic Planning: A Participative Model.

    ERIC Educational Resources Information Center

    Weimer, Don; Jonas, Peter M.

    This paper describes a participative model of strategic planning for use by higher education institutions. First it reviews five principles or assumptions of a participatory model: the person doing the job is the expert; that which is strategic must be validated by the operation; accountability, authority, and information are always equal and…

  14. Strategic Human Resource Development. Symposium.

    ERIC Educational Resources Information Center

    2002

    This document contains three papers on strategic human resource (HR) development. "Strategic HR Orientation and Firm Performance in India" (Kuldeep Singh) reports findings from a study of Indian business executives that suggests there is a positive link between HR policies and practices and workforce motivation and loyalty and…

  15. Strategic Planning Is an Oxymoron

    ERIC Educational Resources Information Center

    Bassett, Patrick F.

    2012-01-01

    The thinking on "strategic thinking" has evolved significantly over the years. In the previous century, the independent school strategy was to focus on long-range planning, blithely projecting 10 years into the future. For decades this worked well enough, but in the late 20th century, independent schools shifted to "strategic planning," with its…

  16. Strategic Planning and Online Learning

    ERIC Educational Resources Information Center

    McLaughlin-Graham, Karen; Berge, Zane L.

    2005-01-01

    Strategic planning is a critical part of sustaining distance education. Through such planning, the organization can solve business problems that involve training and education in an effective and often cost savings manner compared to in-person training efforts. This paper examines the strategic planning process as it relates to sustaining distance…

  17. Army Environmental Cleanup Strategic Plan

    DTIC Science & Technology

    2009-05-01

    Serves an enduring document to guide future strategic plans – Establishes ISO 14001 framework for cleanup; complies w/GPRA  Army Environmental...follow ISO 14001 – Plan - Complete the FY10-11 Strategic Plan – Do - Implement Activities According to the Plan – Check - Evaluate Progress Against the

  18. Strategic Planning for Higher Education.

    ERIC Educational Resources Information Center

    Kotler, Philip; Murphy, Patrick E.

    1981-01-01

    The framework necessary for achieving a strategic planning posture in higher education is outlined. The most important benefit of strategic planning for higher education decision makers is that it forces them to undertake a more market-oriented and systematic approach to long- range planning. (Author/MLW)

  19. Strategic Partnerships in International Development

    ERIC Educational Resources Information Center

    Treat, Tod; Hartenstine, Mary Beth

    2013-01-01

    This chapter provides a framework and recommendations for development of strategic partnerships in a variety of cultural contexts. Additionally, this study elucidates barriers and possibilities in interagency collaborations. Without careful consideration regarding strategic partnerships' approaches, functions, and goals, the ability to…

  20. Strategic Competence and Language Teaching.

    ERIC Educational Resources Information Center

    Rababah, Ghaleb Ahmed

    This paper discusses the notion of communicative competence, particularly strategic competence in English language teaching. Strategic competence refers to the individual's ability to use communication strategies such as paraphrase, circumlocution, literal translation, lexical approximation, and mime to get their message across and to compensate…

  1. Successful strategic planning: creating clarity.

    PubMed

    Adams, Jim

    2005-01-01

    Most healthcare organizations have a strategic plan of some kind. Many of these organizations also have difficulty translating their strategic plan into specific actions that result in successful performance. In the worst cases, this can jeopardize the viability of the organization. The trouble lies in a lack of clarity in what a strategic plan is and what it should do for the organization. This article will answer key questions such as: What is strategy and how does it fit with other commonly used constructs such as mission, vision, and goals? What criteria can be used to determine if something is truly strategic to the organization? What are the phases of the strategy lifecycle? How do approaches for dealing with uncertainty, such as scenario planning, fit with organizational strategic planning? How can a meaningful IT strategy be developed if the organization strategy is lacking? What principles should guide a good IT planning process?

  2. Gene expression profiling with Cre-conditional pseudorabies virus reveals a subset of midbrain neurons that participate in reward circuitry.

    PubMed

    Pomeranz, Lisa E; Ekstrand, Mats I; Latcha, Kaamashri N; Smith, Gregory A; Enquist, Lynn W; Friedman, Jeffrey M

    2017-03-10

    The mesolimbic dopamine pathway receives inputs from numerous regions of the brain as part of a neural system that detects rewarding stimuli and coordinates a behavioral response. The capacity to simultaneously map and molecularly define the components of this complex multisynaptic circuit would thus advance our understanding of the determinants of motivated behavior. To accomplish this, we have constructed pseudorabies virus (PRV) strains in which viral propagation and fluorophore expression are activated only after exposure to Cre recombinase. Once activated in Cre-expressing neurons, the virus serially labels chains of presynaptic neurons. Dual injection of GFP and mCherry tracing viruses simultaneously illuminates nigrostriatal and mesolimbic circuitry and shows no overlap demonstrating that PRV transmission is confined to synaptically connected neurons. In order to molecularly profile mesolimbic dopamine neurons and their presynaptic inputs, we injected Cre-conditional GFP virus into the NAc of (anti-GFP) nanobody-L10 transgenic mice and immunoprecipitated translating ribosomes from neurons infected after retrograde tracing. Analysis of purified RNA revealed an enrichment of transcripts expressed in neurons of the dorsal raphe nuclei and lateral hypothalamus that project to the mesolimbic dopamine circuit. These studies identify important inputs to the mesolimbic dopamine pathway and further show that PRV circuit-directed translating ribosome affinity purification (PRV-Circuit-TRAP) can be broadly applied to identify molecularly defined neurons comprising complex, multisynaptic circuits.SIGNIFICANCE STATEMENTThe mesolimbic dopamine circuit integrates signals from key brain regions in order to detect and respond to rewarding stimuli. To further define this complex multisynaptic circuit, we constructed a panel of Cre recombinase-activated pseudorabies viruses (PRV) that enabled retrograde tracing of neural inputs that terminate on Cre-expressing neurons. Using

  3. Characterization of genetically engineered mouse models carrying Col2a1-cre-induced deletions of Lrp5 and/or Lrp6

    PubMed Central

    Schumacher, Cassie A; Joiner, Danese M; Less, Kennen D; Drewry, Melissa Oosterhouse; Williams, Bart O

    2016-01-01

    Mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 were created and characterized. Mice lacking either gene alone were viable and fertile with normal knee morphology. Mice in which both Lrp5 and Lrp6 were conditionally ablated via Collagen2a1-cre-mediated deletion displayed severe defects in skeletal development during embryogenesis. In addition, adult mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 displayed low bone mass suggesting that the Collagen2a1-cre transgene was active in cells that subsequently differentiated into osteoblasts. In both embryonic skeletal development and establishment of adult bone mass, Lrp5 and Lrp6 carry out redundant functions. PMID:26962465

  4. Strategic cycling: shaking complacency in healthcare strategic planning.

    PubMed

    Begun, J; Heatwole, K B

    1999-01-01

    As the conditions affecting business and healthcare organizations in the United States have become more turbulent and uncertain, strategic planning has decreased in popularity. Strategic planning is criticized for stiffling creative responses to the new marketplace and for fostering compartmentalized organizations, adherence to outmoded strategies, tunnel vision in strategy formulation, and overemphasis on planning to the detriment of implementation. However, effective strategic planning can be a force for mobilizing all the constituents of an organization, creating discipline in pursuit of a goal, broadening an organization's perspective, improving communication among disciplines, and motivating the organization's workforce. It is worthwhile for healthcare organizations to preserve these benefits of strategic planning at the same time recognizing the many sources of turbulence and uncertainty in the healthcare environment. A model of "strategic cycling" is presented to address the perceived shortcomings of traditional strategic planning in a dynamic environment. The cycling model facilitates continuous assessment of the organization's mission/values/vision and primary strategies based on feedback from benchmark analysis, shareholder impact, and progress in strategy implementation. Multiple scenarios and contingency plans are developed in recognition of the uncertain future. The model represents a compromise between abandoning strategic planning and the traditional, linear model of planning based on progress through predetermined stages to a masterpiece plan.

  5. A Strategic Action Plan for Advancing Math and Science Education in New Mexico 2007-2010

    ERIC Educational Resources Information Center

    New Mexico Public Education Department, 2007

    2007-01-01

    This Strategic Action Plan for Advancing Math and Science Education is an initial outline of strategies, actions, measures of progress, resources needed, timelines, and responsible parties. The Plan focuses on these three main goals: (1) increasing student interest, participation, and achievement in math and science; (2) raising public support and…

  6. Insights into the preferential order of strand exchange in the Cre/loxP recombinase system: impact of the DNA spacer flanking sequence and flexibility

    NASA Astrophysics Data System (ADS)

    Abi-Ghanem, Josephine; Samsonov, Sergey A.; Pisabarro, M. Teresa

    2015-03-01

    The Cre/loxP system is widely used as a genetic tool to manipulate DNA. Cre recombinase catalyzes site-specific recombination between 34 bp loxP sites. Each loxP site is recognized by two Cre molecules assuming a cleaving (CreC) and non-cleaving (CreNC) activity. Despite the symmetry in the sequences of the arms of loxP, available biochemical data show strong evidence that the recombination reaction is asymmetric with a preferred strand exchange order. The asymmetry comes from the spacer separating the two sets of palindromic arms of the loxP sequence. However, it remains to be understood how this preferential order is established. We apply computational structure-based methods and perform a thorough detailed analysis of available structural and biochemical information on the Cre/loxP system in order to investigate such asymmetry in the recombination, and we propose a rationale to explain the determinants favoring the strand exchange order. We show that the structural properties of the DNA flanking sequence of the spacer guide the recombination, and we establish the role of residues R118, R121 and K122 from CreC, which contact the spacer region and by clamping the DNA inhibit the cleavage on the second arm of loxP. Our studies give an atomistic insight on the synapsis state of the recombination process in the Cre/loxP system and highlight the importance of the flexibility and other intrinsic properties of the flanking regions of the DNA spacer to establish a preferential strand exchange order.

  7. Conventional Strategic Deterrence

    SciTech Connect

    Latter, A.L.; Martinelli, E.A.; Speed, R.D.

    1992-08-01

    The Bush Administration argues that the US, as the world's only remaining superpower, must be prepared to intervene militarily in regional conflicts. However, the traditional American way of fighting-relying on ground forces with heavy equipment, supported by naval and air forces--could prove too expensive, both monetarily and in terms of expected American casualties, to garner the support of the American public or Congress. This paper argues that the revolution in conventional weaponry demonstrated in the Persian Gulf War opens up the possibility of a new strategy--called Conventional Strategic Deterrence--that could reduce both financial costs and casualties (if it were necessary to implement the strategy) while still being a strong and credible deterrent to regional conflict.

  8. Improved gene amplification by cell-cycle engineering combined with the Cre-loxP system in Chinese hamster ovary cells.

    PubMed

    Matsuyama, Rima; Tsutsui, Tomomi; Lee, Kyoung Ho; Onitsuka, Masayoshi; Omasa, Takeshi

    2015-12-01

    The dihydrofolate reductase gene amplification system is widely used in Chinese hamster ovary (CHO) cells for the industrial production of therapeutic proteins. To enhance the efficiency of conventional gene amplification systems, we previously presented a novel method using cell-cycle checkpoint engineering. Here, we constructed high-producing and stable cells by the conditional expression of mutant cell division cycle 25 homolog B (CDC25B) using the Cre-loxP system. A bispecific antibody-producing CHO DG44-derived cell line was transfected with floxed mutant CDC25B. After inducing gene amplification in the presence of 250 nM methotrexate, mutant CDC25B sequence was removed by Cre recombinase protein expression. Overexpression of the floxed mutant CDC25B significantly enhanced the efficiency of transgene amplification and productivity. Moreover, the specific production rate of the isolated clone CHO Cre-1 and Cre-2 were approximately 11-fold and 15-fold higher than that of mock-transfected clone CHO Mock-S. Chromosomal aneuploidy was increased by mutant CDC25B overexpression, but Cre-1 and Cre-2 did not show any changes in chromosome number during long-term cultivation, as is the case with CHO Mock-S. Our results suggest that high-producing and stable cells can be constructed by conditionally controlling a cell-cycle checkpoint integrated in conventional gene amplification systems.

  9. Simultaneous deletion of floxed genes mediated by CaMKIIα-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Goα.

    PubMed

    Choi, Chan-Il; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung-Soo; Lee, Young-Don; Birnbaumer, Lutz; Suh-Kim, Haeyoung

    2014-05-02

    The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice generated by crossing CaMKIIα-Cre(+/Cre) mice with floxed ROSA26 lacZ reporter (Rosa26(+/stop-lacZ)) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice. Similarly, when double transgenic Gnao(+/f)::CaMKIIα-Cre(+/Cre) mice carrying a floxed Go-alpha gene (Gnao(f/f)) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (Gnao(Δ)) without inheriting the Cre transgene. The Gnao(Δ/Δ) mice closely resembled conventional Go-alpha knockout mice (Gnao(-/-)) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.

  10. 2006 NASA Strategic Plan

    NASA Technical Reports Server (NTRS)

    2006-01-01

    On January 14, 2004, President George W. Bush announced A Renewed Spirit of Discovery: The President's Vision for U.S. Space Exploration, a new directive for the Nation's space program. The fundamental goal of this directive is "to advance U.S. scientific, security, and economic interests through a robust space exploration program." In issuing it, the President committed the Nation to a journey of exploring the solar system and beyond: returning to the Moon in the next decade, then venturing further into the solar system, ultimately sending humans to Mars and beyond. He challenged NASA to establish new and innovative programs to enhance understanding of the planets, to ask new questions, and to answer questions that are as old as humankind. NASA enthusiastically embraced the challenge of extending a human presence throughout the solar system as the Agency's Vision, and in the NASA Authorization Act of 2005, Congress endorsed the Vision for Space Exploration and provided additional guidance for implementation. NASA is committed to achieving this Vision and to making all changes necessary to ensure success and a smooth transition. These changes will include increasing internal collaboration, leveraging personnel and facilities, developing strong, healthy NASA Centers,a nd fostering a safe environment of respect and open communication for employees at all levels. NASA also will ensure clear accountability and solid program management and reporting practices. Over the next 10 years, NASA will focus on six Strategic Goals to move forward in achieving the Vision for Space Exploration. Each of the six Strategic Goals is clearly defined and supported by multi-year outcomes that will enhance NASA's ability to measure and report Agency accomplishments in this quest.

  11. Successful Proposals, Fund Raising, and Project Leadership

    NASA Astrophysics Data System (ADS)

    Ainsbury, Elizabeth A.; Cassidy, Amy; Downey, Frances; Hartline, Beverly K.; Lin, J. Grace; McCray, Christophe; Dawson, Silvina Ponce

    2009-04-01

    A brief description of the Fundraising, Grant Writing, and Project Leadership workshop at the Third IUPAP International Conference of Women in Physics is presented. The workshop's purpose was to assist delegates in developing skills in successful proposal writing, fund raising, and project leadership. Handouts and talks were given with information on funding sources in different regions. Talks on grant writing and leadership by Beverly Hartline and J. Grace Lin led to active participation and brainstorming discussions. Based on the discussions a list of recommendations is outlined in this paper. This information is also available at the IUPAP Working Group of Women in Physics website (http://wgwip.df.uba.ar).

  12. The Value of Strategic Partnerships

    SciTech Connect

    Gould, Josh; Narayan, Amit; McNutt, Ty

    2015-02-10

    Strong strategic partnerships can be the difference between those technologies that only achieve success in the lab and those that actually break into the marketplace. Two ARPA-E awardees—AutoGrid and APEI—have forged strategic partnerships that have positioned their technologies to achieve major success in the market. This video features remarks from ARPA-E Technology-to-Market Advisor Josh Gould and interviews with technologists at AutoGrid and APEI, who each tell the story of how their company leveraged relationships with strategic partners to broaden their customer base and bring their technology to life.

  13. The Value of Strategic Partnerships

    ScienceCinema

    Gould, Josh; Narayan, Amit; McNutt, Ty

    2016-07-12

    Strong strategic partnerships can be the difference between those technologies that only achieve success in the lab and those that actually break into the marketplace. Two ARPA-E awardees—AutoGrid and APEI—have forged strategic partnerships that have positioned their technologies to achieve major success in the market. This video features remarks from ARPA-E Technology-to-Market Advisor Josh Gould and interviews with technologists at AutoGrid and APEI, who each tell the story of how their company leveraged relationships with strategic partners to broaden their customer base and bring their technology to life.

  14. Preserving the ABM treaty: a critique of the Reagan Strategic Defense Initiative

    SciTech Connect

    Drell, Sidney D.; Farley, Philip J.; Holloway, David

    1994-10-01

    President Reagan's Strategic Defense Initiative (SDI) proposal for long-term research and development on a leak-proof defense capability conflicts with the past 15 years of Soviet-American strategic relations and arms control negotiations. It represents a unilateral change, and will elicit a Soviet response based on how the Soviets think their own security may be affected. Whether the SDI transcends or enhances defense, it raises basic issues involving the US-Soviet strategic relationship. Emphasizing the central bearing of the Anti-Ballistic Missile (ABM) Treaty of 1972 the author reviews these issues and questions about the technical and strategic grounds of the proposal. Three recommendations are to limit the SDI program to a search for scientific possibilities, to consult with the Soviets on the ABM Treaty, and to recognize that the ABM Treaty will enhance security more than the SDI. 58 references.

  15. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  16. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  17. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  18. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  19. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  20. Guidelines for strategic information planning.

    PubMed

    Lederer, A L; Sethi, V

    1991-01-01

    Developing a strategic information plan is an essential step for companies that want to realize the advantages of today's information technology. This article discusses ways to avoid the pitfalls that frequently occur during stages of information planning.

  1. Generation of a Transgenic Mouse for Colorectal Cancer Research with Intestinal Cre-Expression Limited to the Large Intestine

    PubMed Central

    Xue, Yingben; Johnson, Robert; DeSmet, Marsha; Snyder, Paul W.; Fleet, James C.

    2010-01-01

    Genetically modified mice have been used for colon cancer research but findings from these models are confounded by expression of cancer in multiple organs. We sought to create a transgenic mouse with Cre recombinase (Cre) expression limited to the epithelial cells of the large intestine and use this model to study colon cancer driven by adenomatosis polyposis coli (APC) gene inactivation. A promoter/enhancer from the mouse carbonic anhydrase I gene was used to generate a Cre expressing transgenic mouse (CAC). After characterizing transgene expression and distribution, CAC mice were crossed to APC580S mice to generate mice with APC inactivation at one (CAC; APC580S/+) or both alleles (CAC; APC580S/580S). Transgene expression was limited to the epithelial cells of the cecum and colon, extended from the crypt base to the luminal surface, and was expressed in approximately 15% of the crypts. No abnormal gross phenotype was seen in 3 or 6 wk CAC; APC580S/+ mice but CAC; APC580S/580S mice had significant mucosal hyperplasia in the colon at 3 wk that developed into tumors by 6 wk. By 10 wk, 20% of CAC; APC580S/+ mice developed adenomatous lesions in the distal colon (3.0±0.4 mm, 1.1 per mouse). Dextran sulfate sodium treatment increased the incidence and number of tumors and this occurred predominantly in distal colon. Our new model has improved features for colon cancer research i.e. transgene expression is limited to the epithelium of the large bowel with normal cells found next to genetically modified cells. PMID:20663863

  2. Novel migrating mouse neural crest cell assay system utilizing P0-Cre/EGFP fluorescent time-lapse imaging

    PubMed Central

    2011-01-01

    Background Neural crest cells (NCCs) are embryonic, multipotent stem cells. Their long-range and precision-guided migration is one of their most striking characteristics. We previously reported that P0-Cre/CAG-CAT-lacZ double-transgenic mice showed significant lacZ expression in tissues derived from NCCs. Results In this study, by embedding a P0-Cre/CAG-CAT-EGFP embryo at E9.5 in collagen gel inside a culture glass slide, we were able to keep the embryo developing ex vivo for more than 24 hours; this development was with enough NCC fluorescent signal intensity to enable single-cell resolution analysis, with the accompanying NCC migration potential intact and with the appropriate NCC response to the extracellular signal maintained. By implantation of beads with absorbed platelet-derived growth factor-AA (PDGF-AA), we demonstrated that PDGF-AA acts as an NCC-attractant in embryos. We also performed assays with NCCs isolated from P0-Cre/CAG-CAT-EGFP embryos on culture plates. The neuromediator 5-hydroxytryptamine (5-HT) has been known to regulate NCC migration. We newly demonstrated that dopamine, in addition to 5-HT, stimulated NCC migration in vitro. Two NCC populations, with different axial levels of origins, showed unique distribution patterns regarding migration velocity and different dose-response patterns to both 5-HT and dopamine. Conclusions Although avian species predominated over the other species in the NCC study, our novel system should enable us to use mice to assay many different aspects of NCCs in embryos or on culture plates, such as migration, division, differentiation, and apoptosis. PMID:22070366

  3. Developing Future Strategic Logistics Leaders

    DTIC Science & Technology

    2013-03-01

    and our focus on tactical level lessons learned has 12 skewed our emphasis in PME to a very narrow band of excellence. As such our current...A partnership between senior logistics leaders, PME developers, and personnel managers is essential to constructing and maintaining strategic...short term and inconsistent PME system that does not produce strategic logistic leaders. The Logistics Corps, sustainment branches, LOO lead for leader

  4. Strategic planning: myth or reality.

    PubMed

    Davidson, D

    2000-06-01

    The reality of strategic planning often falls prey to the unplanned external or internal forces at work in health care. A strategic plan may sit on the shelf as though it were an end unto itself. This article discusses what might need to happen differently to change the outcome of planning generally and describes some of the strengths and weaknesses of planning at the system level.

  5. Tamoxifen Activation of Cre-Recombinase Has No Persisting Effects on Adult Neurogenesis or Learning and Anxiety

    PubMed Central

    Rotheneichner, Peter; Romanelli, Pasquale; Bieler, Lara; Pagitsch, Sebastian; Zaunmair, Pia; Kreutzer, Christina; König, Richard; Marschallinger, Julia; Aigner, Ludwig; Couillard-Després, Sébastien

    2017-01-01

    Adult neurogenesis is a tightly regulated process continuously taking place in the central nervous system of most mammalian species. In neuroscience research, transgenic animals bearing the tamoxifen-inducible CreERT2-Lox system are widely used. In this study, we made use of a Nestin-CreERT2/R26R-YFP transgenic mouse model in which the CreERT2 activates the expression of YFP in multipotent neural stem cells upon tamoxifen application. Humoral factors, such as the levels of estrogens, have been reported to affect the hippocampal neurogenesis. The application of tamoxifen, a mixed agonist/antagonist of the estrogen receptor that permeates the blood-brain-barrier, could thus influence adult neurogenesis. Although the functions of adult neurogenesis are yet to be fully deciphered, a reciprocal interaction between rates of neurogenesis on the one hand and learning and mood regulation on the other hand, has been suggested. The impact of tamoxifen on neurogenesis and behavior was therefore addressed following five daily applications according to the open field test, the elevated plus maze, and Morris water maze. In addition, the impact of short-term tamoxifen application on progenitor cell proliferation, morphology, and fate in the neurogenic niche of the dentate gyrus were investigated. Finally, the influence of the route of administration (oral vs. intra-peritoneal) and gender-specific response were scrutinized. The sub-acute analysis did neither reveal significant differences in behavior, such as voluntary motor activity, anxiety behavior, and spatial learning, nor in cell proliferation, cell survival, dendritic arborization or maturation rate within the dentate gyrus between saline solution-, corn oil-, and tamoxifen-treated groups. Finally, neither the route of application, nor the gender of treated mice influenced the response to tamoxifen. We conclude that short tamoxifen treatments used to activate the CreERT2 system in transgenic mouse models does not have a

  6. Tamoxifen Activation of Cre-Recombinase Has No Persisting Effects on Adult Neurogenesis or Learning and Anxiety.

    PubMed

    Rotheneichner, Peter; Romanelli, Pasquale; Bieler, Lara; Pagitsch, Sebastian; Zaunmair, Pia; Kreutzer, Christina; König, Richard; Marschallinger, Julia; Aigner, Ludwig; Couillard-Després, Sébastien

    2017-01-01

    Adult neurogenesis is a tightly regulated process continuously taking place in the central nervous system of most mammalian species. In neuroscience research, transgenic animals bearing the tamoxifen-inducible CreER(T2)-Lox system are widely used. In this study, we made use of a Nestin-CreER(T2)/R26R-YFP transgenic mouse model in which the CreER(T2) activates the expression of YFP in multipotent neural stem cells upon tamoxifen application. Humoral factors, such as the levels of estrogens, have been reported to affect the hippocampal neurogenesis. The application of tamoxifen, a mixed agonist/antagonist of the estrogen receptor that permeates the blood-brain-barrier, could thus influence adult neurogenesis. Although the functions of adult neurogenesis are yet to be fully deciphered, a reciprocal interaction between rates of neurogenesis on the one hand and learning and mood regulation on the other hand, has been suggested. The impact of tamoxifen on neurogenesis and behavior was therefore addressed following five daily applications according to the open field test, the elevated plus maze, and Morris water maze. In addition, the impact of short-term tamoxifen application on progenitor cell proliferation, morphology, and fate in the neurogenic niche of the dentate gyrus were investigated. Finally, the influence of the route of administration (oral vs. intra-peritoneal) and gender-specific response were scrutinized. The sub-acute analysis did neither reveal significant differences in behavior, such as voluntary motor activity, anxiety behavior, and spatial learning, nor in cell proliferation, cell survival, dendritic arborization or maturation rate within the dentate gyrus between saline solution-, corn oil-, and tamoxifen-treated groups. Finally, neither the route of application, nor the gender of treated mice influenced the response to tamoxifen. We conclude that short tamoxifen treatments used to activate the CreER(T2) system in transgenic mouse models does not

  7. Isolation and Culture of Adult Intestinal, Gastric, and Liver Organoids for Cre-recombinase-Mediated Gene Deletion.

    PubMed

    Flanagan, Dustin J; Schwab, Renate H M; Tran, Bang M; Phesse, Toby J; Vincan, Elizabeth

    2016-10-05

    The discovery of Lgr5 as a marker of adult stem cells meant that stem cell populations could be purified and studied in isolation. Importantly, when cultured under the appropriate conditions these stem cells form organoids in tissue culture that retain many features of the tissue of origin. The organoid cultures are accessible to genetic and biochemical manipulation, bridging the gap between in vivo mouse models and conventional tissue culture. Here we describe robust protocols to establish organoids from gastrointestinal tissues (stomach, intestine, liver) and Cre-recombinase mediated gene manipulation in vitro.

  8. The Pseudomonas aeruginosa CreBC Two-Component System Plays a Major Role in the Response to β-Lactams, Fitness, Biofilm Growth, and Global Regulation

    PubMed Central

    Zamorano, Laura; Juan, Carlos; Mulet, Xavier; Blázquez, Jesús

    2014-01-01

    Pseudomonas aeruginosa is a ubiquitous versatile environmental microorganism with a remarkable ability to grow under diverse environmental conditions. Moreover, P. aeruginosa is responsible for life-threatening infections in immunocompromised and cystic fibrosis patients, as the extraordinary capacity of this pathogen to develop antimicrobial resistance dramatically limits our therapeutic arsenal. Its large genome carries an outstanding number of genes belonging to regulatory systems, including multiple two-component sensor-regulator systems that modulate the response to the different environmental stimuli. Here, we show that one of two systems, designated CreBC (carbon source responsive) and BlrAB (β-lactam resistance), might be of particular relevance. We first identified the stimuli triggering the activation of the CreBC system, which specifically responds to penicillin-binding protein 4 (PBP4) inhibition by certain β-lactam antibiotics. Second, through an analysis of a large comprehensive collection of mutants, we demonstrate an intricate interconnection between the CreBC system, the peptidoglycan recycling pathway, and the expression of the concerning chromosomal β-lactamase AmpC. Third, we show that the CreBC system, and particularly its effector inner membrane protein CreD, plays a major role in bacterial fitness and biofilm development, especially in the presence of subinhibitory concentrations of β-lactams. Finally, global transcriptomics reveals broad regulatory functions of CreBC in basic physiological aspects, particularly anaerobic respiration, in both the presence and absence of antibiotics. Therefore, the CreBC system is envisaged as a potentially interesting target for improving the efficacy of β-lactams against P. aeruginosa infections. PMID:24936599

  9. Maintenance Process Strategic Analysis

    NASA Astrophysics Data System (ADS)

    Jasiulewicz-Kaczmarek, M.; Stachowiak, A.

    2016-08-01

    The performance and competitiveness of manufacturing companies is dependent on the availability, reliability and productivity of their production facilities. Low productivity, downtime, and poor machine performance is often linked to inadequate plant maintenance, which in turn can lead to reduced production levels, increasing costs, lost market opportunities, and lower profits. These pressures have given firms worldwide the motivation to explore and embrace proactive maintenance strategies over the traditional reactive firefighting methods. The traditional view of maintenance has shifted into one of an overall view that encompasses Overall Equipment Efficiency, Stakeholders Management and Life Cycle assessment. From practical point of view it requires changes in approach to maintenance represented by managers and changes in actions performed within maintenance area. Managers have to understand that maintenance is not only about repairs and conservations of machines and devices, but also actions striving for more efficient resources management and care for safety and health of employees. The purpose of the work is to present strategic analysis based on SWOT analysis to identify the opportunities and strengths of maintenance process, to benefit from them as much as possible, as well as to identify weaknesses and threats, so that they could be eliminated or minimized.

  10. Strategic defense initiative

    SciTech Connect

    Nichols, J.P.

    1985-01-01

    The Engineering Technology Division has a leading role, including that of program management, in a major new programmatic thrust of the Oak Ridge National Laboratory that is in support of the national Strategic Defense Initiative (SDI). It is appropriate for the Laboratory to become significantly involved in the program because several of the most promising SDI technologies are in areas for which ORNL (together with Y-12 and K-25) have strong capabilities and significant resources. The initial ORNL work in support of the SDI program is focused on three technologies in which ORNL has extensive experience and traditionally strong research and development programs: (1) space nuclear power, (2) flywheel energy storage, and (3) neutral particle beams. The space nuclear program will utilize our capabilities in areas such as refractory materials, high-temperature alkali metal systems, shielding, and instrumentation. Space nuclear reactors capable of supplying multimegawatt levels of electrical power on a continuous and long-term basis are envisioned to be required for a variety of SDI surveillance satellites and space-borne weapons platforms. The feasibility of an alkali metal Rankine power conversion cycle, which has promise of providing high power with a very low system mass, is planned for study.

  11. CINT 2020 Strategic Plan

    SciTech Connect

    Shinn, Neal D.

    2014-12-01

    CINT’s role is to enable world-leading science towards realizing these benefits and our strategic objectives describe what is needed to deliver on this promise. As a vibrant partnership between Los Alamos National Laboratory (LANL) and Sandia National Laboratories (SNL), CINT leverages the unmatched scientific and engineering expertise of our host DOE Laboratories in an Office of Science open-access user facility to benefit hundreds of researchers annually. We have world-leading scientific expertise in four thrust areas, as described in section 1, and specialized capabilities to create, characterize and understand nanomaterials in increasingly complex integrated environments. Building upon these current strengths, we identify some of the capabilities and expertise that the nanoscience community will need in the future and that CINT is well positioned to develop and offer as a user facility. These include an expanding portfolio of our signature Discovery Platforms that can be used alone or as sophisticated “experiments within an experiment”; novel synthetic approaches for exquisitely heterostructured nanowires, nanoparticles and quasi-two-dimensional materials; ultra-high resolution spectroscopic techniques of nanomaterial dynamics; in situ microscopies that provide realtime, spatially-resolved structure/property information for increasingly complex materials systems; advanced simulation techniques for integrated nanomaterials; and multi-scale theory for interfaces and dynamics.

  12. The need for strategic tax planning among nonprofit hospitals.

    PubMed

    Smith, Pamela C

    2005-01-01

    Strategic tax planning issues are important to the nonprofit health care sector, despite its philanthropic mission. The consolidation of the industry has led management to fight for resources and develop alternative strategies for raising money. When management evaluates alternative collaborative structures to increase efficiency, the impact on governance structures must also be considered. The increased governmental scrutiny of joint ventures within the health care sector warrants management's attention as well. The financial incentives must be considered, along with the various tax policy implications of cross-sector collaborations.

  13. Alternate Sources of Funding: Scholarship Fund Raising. and Anne Arundel Community College Scholarship Fund Raising Campaign.

    ERIC Educational Resources Information Center

    Weinberg, Barry M.

    This two-part paper provides guidance on alternative sources of funding for college scholarships and describes Anne Arundel Community College's (AACC's) Scholarship Fund Raising Campaign. First, an outline is presented, covering: (1) prerequisites to fundraising (i.e., leadership, sense of direction, commitment, and a plan for identifying,…

  14. Against Raising Hope of Raising the Dead: Contra Moody and Kubler-Ross.

    ERIC Educational Resources Information Center

    Vicchio, Stephen J.; And Others

    1979-01-01

    Kubler-Ross and Moody have made assertions about survival after death. They argued that the subjects were not dead, but in the process of dying. An alternative explanation to this "glimpse of the afterlife" approach is offered. Other theological objections are raised to the Moody/Kubler-Ross approach. (Author)

  15. Fund-Raising While You Shop: A Look at Fund-Raising Sites on the Internet.

    ERIC Educational Resources Information Center

    Seydel, Angela

    2000-01-01

    Explains how school fund-raising can be done over the Internet through electronic commerce. Discusses online retailers who donate percentage of sales to schools who have registered; Web sites that coordinate online retailers for schools, who also take a percentage of the sales; loss of revenue for local businesses; and marketing. (LRW)

  16. Nanofaceted C/Re(1121): fabrication, structure, and template for synthesizing nanostructured model Pt electrocatalyst for hydrogen evolution reaction.

    PubMed

    Yang, Xiaofang; Koel, Bruce E; Wang, Hao; Chen, Wenhua; Bartynski, Robert A

    2012-02-28

    We report the first observation of carbon-induced nanofaceting of a Re single crystal and its application in synthesizing a nanostructured model Pt electrocatalyst investigated using multiple surface science techniques, including low-energy electron diffraction, Auger electron spectroscopy, X-ray photoelectron spectroscopy, low-energy ion scattering, and scanning tunneling microscopy, combined with electrochemical reaction measurements. Upon annealing in acetylene at 700 K followed by annealing in vacuum at 1100 K, an initially planar Re(112̅1) surface becomes completely faceted and covered with three-sided nanopyramids exposing (011̅1), (101̅1), and (112̅0) faces. Using the faceted C/Re(112̅1) surface as a template, we have successfully fabricated a nanostructured Pt monolayer (ML) electrocatalyst. The Pt ML supported on the C/Re nanotemplate exhibits higher activity for the hydrogen evolution reaction than Pt(111). This is the first application of faceted metal surfaces as templates for synthesis of nanoscale model electrocatalyst with well-defined (facet) surface structure and controlled (facet) size on the nanometer scale, illustrating the potential for future studies of nanostructured bimetallic systems relevant to electrocatalytic reactions.

  17. A high throughput Cre-lox activated viral membrane fusion assay identifies pharmacological inhibitors of HIV entry.

    PubMed

    Esposito, Anthony M; Cheung, Pamela; Swartz, Talia H; Li, Hongru; Tsibane, Tshidi; Durham, Natasha D; Basler, Christopher F; Felsenfeld, Dan P; Chen, Benjamin K

    2016-03-01

    Enveloped virus entry occurs when viral and cellular membranes fuse releasing particle contents into the target cell. Human immunodeficiency virus (HIV) entry occurs by cell-free virus or virus transferred between infected and uninfected cells through structures called virological synapses. We developed a high-throughput cell-based assay to identify small molecule inhibitors of cell-free or virological synapse-mediated entry. An HIV clone carrying Cre recombinase as a Gag-internal gene fusion releases active Cre into cells upon viral entry activating a recombinatorial gene switch changing dsRed to GFP-expression. A screen of a 1998 known-biological profile small molecule library identified pharmacological HIV entry inhibitors that block both cell-free and cell-to-cell infection. Many top hits were noted as HIV inhibitors in prior studies, but not previously recognized as entry antagonists. Modest therapeutic indices for simvastatin and nigericin were observed in confirmatory HIV infection assays. This robust assay is adaptable to study HIV and heterologous viral pseudotypes.

  18. Stereotaxic microinjection of viral vectors expressing Cre recombinase to study the role of target genes in cocaine conditioned place preference.

    PubMed

    Schierberl, Kathryn C; Rajadhyaksha, Anjali M

    2013-07-30

    Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

  19. An araC-controlled bacterial cre expression system to produce DNA minicircle vectors for nuclear and mitochondrial gene therapy.

    PubMed

    Bigger, B W; Tolmachov, O; Collombet, J M; Fragkos, M; Palaszewski, I; Coutelle, C

    2001-06-22

    The presence of CpG motifs and their associated sequences in bacterial DNA causes an immunotoxic response following the delivery of these plasmid vectors into mammalian hosts. We describe a biotechnological approach to the elimination of this problem by the creation of a bacterial cre recombinase expression system, tightly controlled by the arabinose regulon. This permits the Cre-mediated and -directed excision of the entire bacterial vector sequences from plasmid constructs to create supercoiled gene expression minicircles for gene therapy. Minicircle yields using standard culture volumes are sufficient for most in vitro and in vivo applications whereas minicircle expression in vitro is significantly increased over standard plasmid transfection. By the simple expedient of removing the bacterial DNA complement, we significantly reduce the size and CpG content of these expression vectors, which should also reduce DNA-induced inflammatory responses in a dose-dependent manner. We further describe the generation of minicircle expression vectors for mammalian mitochondrial gene therapy, for which no other vector systems currently exist. The removal of bacterial vector sequences should permit appropriate transcription and correct transcriptional cleavage from the mitochondrial minicircle constructs in a mitochondrial environment and brings the realization of mitochondrial gene therapy a step closer.

  20. Improved transgene integration into the Chinese hamster ovary cell genome using the Cre-loxP system.

    PubMed

    Inao, Takanori; Kawabe, Yoshinori; Yamashiro, Takuro; Kameyama, Yujiro; Wang, Xue; Ito, Akira; Kamihira, Masamichi

    2015-07-01

    Genetic engineering of cellular genomes has provided useful tools for biomedical and pharmaceutical studies such as the generation of transgenic animals and producer cells of biopharmaceutical proteins. Gene integration using site-specific recombinases enables precise transgene insertion into predetermined genomic sites if the target site sequence is introduced into a specific chromosomal locus. We previously developed an accumulative site-specific gene integration system (AGIS) using Cre and mutated loxPs. The system enabled the repeated integration of multiple transgenes into a predetermined locus of a genome. In this study, we explored applicable mutated loxP pairs for AGIS to improve the integration efficiency. The integration efficiencies of 52 mutated loxP sequences, including novel sequences, were measured using an in vitro evaluation system. Among mutated loxP pairs that exhibited a high integration efficiency, the applicability of the selected pairs to AGIS was confirmed for transgene integration into the Chinese hamster ovary cell genome. The newly found mutated loxP pairs should be useful for Cre-mediated integration of transgenes and AGIS.

  1. Reversibility of the Snail-induced epithelial-mesenchymal transition revealed by the Cre-loxP system.

    PubMed

    Ozawa, Masayuki; Kobayashi, Wakako

    2015-03-13

    The epithelial-mesenchymal transition (EMT), a key process in the tumor metastatic cascade, is characterized by the loss of cell-cell junctions and cell polarity, as well as the acquisition of migratory and invasive properties. Snail is an EMT-inducer whose expression in several different epithelial cells, e.g., Madin-Darby canine kidney (MDCK), leads to EMT. To further understand EMT induced by Snail expression, the Cre-loxP site-specific recombination system was used to investigate its reversibility. Transfection of MDCK cells with loxP-flanked Snail (Snail-loxP) resulted in EMT induction, which included the acquisition of a spindle-shaped fibroblastic morphology, the downregulation of epithelial markers, and the upregulation of mesenchymal markers. DNA methylation of the E-cadherin promoter, which often occurs during E-cadherin downregulation, was not observed in Snail+ cells. After Cre-mediated excision of Snail-loxP, the cells reacquired an epithelial morphology, upregulated epithelial markers, and downregulated mesenchymal markers. Thus, EMT induced by Snail expression was reversible.

  2. Children's strategic theory of mind.

    PubMed

    Sher, Itai; Koenig, Melissa; Rustichini, Aldo

    2014-09-16

    Human strategic interaction requires reasoning about other people's behavior and mental states, combined with an understanding of their incentives. However, the ontogenic development of strategic reasoning is not well understood: At what age do we show a capacity for sophisticated play in social interactions? Several lines of inquiry suggest an important role for recursive thinking (RT) and theory of mind (ToM), but these capacities leave out the strategic element. We posit a strategic theory of mind (SToM) integrating ToM and RT with reasoning about incentives of all players. We investigated SToM in 3- to 9-y-old children and adults in two games that represent prevalent aspects of social interaction. Children anticipate deceptive and competitive moves from the other player and play both games in a strategically sophisticated manner by 7 y of age. One game has a pure strategy Nash equilibrium: In this game, children achieve equilibrium play by the age of 7 y on the first move. In the other game, with a single mixed-strategy equilibrium, children's behavior moved toward the equilibrium with experience. These two results also correspond to two ways in which children's behavior resembles adult behavior in the same games. In both games, children's behavior becomes more strategically sophisticated with age on the first move. Beyond the age of 7 y, children begin to think about strategic interaction not myopically, but in a farsighted way, possibly with a view to cooperating and capitalizing on mutual gains in long-run relationships.

  3. Transcriptional regulation of the mouse alpha A-crystallin gene: activation dependent on a cyclic AMP-responsive element (DE1/CRE) and a Pax-6-binding site.

    PubMed Central

    Cvekl, A; Kashanchi, F; Sax, C M; Brady, J N; Piatigorsky, J

    1995-01-01

    Two cis-acting promoter elements (-108 to -100 and -49 to -33) of the mouse alpha A-crystallin gene, which is highly expressed in the ocular lens, were studied. Here we show that DE1 (-108 to -100; 5'TGACGGTG3'), which resembles the consensus cyclic AMP (cAMP)-responsive element sequence (CRE; 5'TGACGT[A/C][A/G]3'), behaves like a functional CRE site. Transfection experiments and electrophoretic mobility shift assays (EMSAs) using site-specific mutations correlated a loss of function with deviations from the CRE consensus sequence. Results of EMSAs in the presence of antisera against CREB, delta CREB, and CREM were consistent with the binding of CREB-like proteins to the DE1 sequence. Stimulation of alpha A-crystallin promoter activity via 8-bromo-cAMP, forskolin, or human T-cell leukemia virus type I Tax1 in transfections and reduction of activity of this site in cell-free transcription tests by competition with the somatostatin CRE supported the idea that DE1 is a functional CRE. Finally, Pax-6, a member of the paired-box family of transcription factors, activated the mouse alpha A-crystallin promoter in cotransfected COP-8 fibroblasts and bound to the -59 to -29 promoter sequence in EMSAs. These data provide evidence for a synergistic role of Pax-6 and CREB-like proteins for high expression of the mouse alpha A-crystallin gene in the lens. PMID:7823934

  4. Generation of knock-in mice that express nuclear enhanced green fluorescent protein and tamoxifen-inducible Cre recombinase in the notochord from Foxa2 and T loci.

    PubMed

    Imuta, Yu; Kiyonari, Hiroshi; Jang, Chuan-Wei; Behringer, Richard R; Sasaki, Hiroshi

    2013-03-01

    The node and the notochord are important embryonic signaling centers that control embryonic pattern formation. Notochord progenitor cells present in the node and later in the posterior end of the notochord move anteriorly to generate the notochord. To understand the dynamics of cell movement during notochord development and the molecular mechanisms controlling this event, analyses of cell movements using time-lapse imaging and conditional manipulation of gene activities are required. To achieve this goal, we generated two knock-in mouse lines that simultaneously express nuclear enhanced green fluorescent protein (EGFP) and tamoxifen-inducible Cre, CreER(T2) , from two notochord gene loci, Foxa2 and T (Brachury). In Foxa2(nEGFP-CreERT2/+) and T(nEGFP-CreERT2/+) embryos, nuclei of the Foxa2 or T-expressing cells, which include the node, notochord, and endoderm (Foxa2) or wide range of posterior mesoderm (T), were labeled with EGFP at intensities that can be used for live imaging. Cre activity was also induced in cells expressing Foxa2 and T 1 day after tamoxifen administration. These mice are expected to be useful tools for analyzing the mechanisms of notochord development.

  5. Generation of brain tumours in mice by Cre-mediated recombination of neural progenitors in situ with the tamoxifen metabolite endoxifen.

    PubMed

    Benedykcinska, Anna; Ferreira, Andreia; Lau, Joanne; Broni, Jessica; Richard-Loendt, Angela; Henriquez, Nico V; Brandner, Sebastian

    2016-02-01

    Targeted cell- or region-specific gene recombination is widely used in the functional analysis of genes implicated in development and disease. In the brain, targeted gene recombination has become a mainstream approach to study neurodegeneration or tumorigenesis. The use of the Cre-loxP system to study tumorigenesis in the adult central nervous system (CNS) can be limited, when the promoter (such as GFAP) is also transiently expressed during development, which can result in the recombination of progenies of different lineages. Engineering of transgenic mice expressing Cre recombinase fused to a mutant of the human oestrogen receptor (ER) allows the circumvention of transient developmental Cre expression by inducing recombination in the adult organism. The recombination of loxP sequences occurs only in the presence of tamoxifen. Systemic administration of tamoxifen can, however, exhibit toxicity and might also recombine unwanted cell populations if the promoter driving Cre expression is active at the time of tamoxifen administration. Here, we report that a single site-specific injection of an active derivative of tamoxifen successfully activates Cre recombinase and selectively recombines tumour suppressor genes in neural progenitor cells of the subventricular zone in mice, and we demonstrate its application in a model for the generation of intrinsic brain tumours.

  6. Expression pattern of cellulolytic and xylanolytic genes regulated by transcriptional factors XYR1 and CRE1 are affected by carbon source in Trichoderma reesei.

    PubMed

    Castro, Lilian dos Santos; Antoniêto, Amanda Cristina Campos; Pedersoli, Wellington Ramos; Silva-Rocha, Rafael; Persinoti, Gabriela F; Silva, Roberto Nascimento

    2014-03-01

    Trichoderma reesei is the most important fungus for the industrial production of enzymes to biomass deconstruction. Most of the genes encoding cellulases and hemicellulases are regulated by the transcription factors CRE1 and XYR1. In this work, the regulation of 22 genes of cellulases and xylanases by these transcription factors was investigated under three different carbon sources. Analysis of gene expression and enzymatic profiles of CMCase, β-glucosidase, and xylanases showed different regulation that was depended of the carbon source in both Δxyr1 and Δcre1 mutants. In the presence of glucose, the majority of genes evaluated (82%) showed increased expression levels in the Δcre1 mutant compared to the parental QM9414 strain. In the Δxyr1 mutant, it was observed that expression of cellulase and xylanase genes was reduced compared to the parental QM9414 strain, when cultured in the presence of cellulose or sophorose. Interesting, in the presence of glucose, approximately 60% of the analyzed genes had increased expression in the Δxyr1 mutant compared to parental strain. Furthermore, no correlation between gene expression and the number of putative binding sites of XYR1 and CRE1 to promoter region of cellulolytic and xylanolytic studied genes was observed. Therefore, these results demonstrated that the regulation of cellulase and xylanase by the transcription factors CRE1 and XYR1 is influenced by different carbon sources.

  7. Cre/loxP plus BAC”: a strategy for direct cloning of large DNA fragment and its applications in Photorhabdus luminescens and Agrobacterium tumefaciens

    PubMed Central

    Hu, Shengbiao; Liu, Zhengqiang; Zhang, Xu; Zhang, Guoyong; Xie, Yali; Ding, Xuezhi; Mo, Xiangtao; Stewart, A. Francis; Fu, Jun; Zhang, Youming; Xia, Liqiu

    2016-01-01

    Heterologous expression has been proven to be a valid strategy for elucidating the natural products produced by gene clusters uncovered by genome sequencing projects. Efforts have been made to efficiently clone gene clusters directly from genomic DNA and several approaches have been developed. Here, we present an alternative strategy based on the site-specific recombinase system Cre/loxP for direct cloning gene clusters. A type three secretion system (T3SS) gene cluster (~32 kb) from Photorhabdus luminescens TT01 and DNA fragment (~78 kb) containing the siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58 have been successfully cloned into pBeloBAC11 with “Cre/loxP plus BAC” strategy. Based on the fact that Cre/loxP system has successfully used for genomic engineering in a wide range of organisms, we believe that this strategy could be widely used for direct cloning of large DNA fragment. PMID:27364376

  8. Current Protocols in Mouse Biology Tissue-specific regulation of oncogene expression using Cre-inducible ROSA26 knock-in transgenic mice

    PubMed Central

    Carofino, Brandi L.; Justice, Monica J.

    2015-01-01

    Cre-inducible mouse models are often utilized for the spatial and temporal expression of oncogenes. With the wide number of Cre recombinase lines available, inducible transgenesis represents a tractable approach to achieve discrete oncogene expression. Here, we describe a protocol for targeting Cre-inducible genes using a loxP-STOP-loxP approach to the ubiquitously expressed ROSA26 locus. Gene targeting provides several advantages over standard transgenic techniques, including a known site of integration and previously characterized pattern of expression. Historically, an inherent instability of ROSA26 targeting vectors has hampered the efficiency of developing ROSA26 knock-in lines. In this protocol, we provide individual steps for utilizing Gateway recombination for cloning, and detailed instructions for screening targeted ES cell clones. By following this protocol, one can achieve germline transmission of a ROSA26 knock-in line within several months. PMID:26069083

  9. Verb-raising and Numeral Classifiers in Japanese: Incompatible Bedfellows.

    ERIC Educational Resources Information Center

    Fukushima, Kazuhiko

    2003-01-01

    Examines verb raising in Japanese and looks at Koizumi's (2000) evidence for verb-raising based on data involving, among other things, numeral classifiers. Demonstrates that Koizumi's evidence based on numeral classifiers does not support his claim that verb-raising occurs in Japanese. (Author/VWL)

  10. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  11. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  12. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  13. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  14. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  15. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  16. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  17. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  18. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  19. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  20. 29 CFR 780.616 - Operations included in raising livestock.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Operations included in raising livestock. 780.616 Section... Employment in Agriculture and Livestock Auction Operations Under the Section 13(b)(13) Exemption Requirements for Exemption § 780.616 Operations included in raising livestock. Raising livestock includes...

  1. Complete fund-raising software systems vs. donor databases.

    PubMed

    2001-10-01

    When evaluating fund-raising software packages, it is important to understand the distinctions between them. Not all fund-raising software is created equal. In fact, there is a whole new breed of fund-raising software that has just entered the market.

  2. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 3 2011-01-01 2011-01-01 false Farm-raised fish. 60.106 Section 60.106 Agriculture... FOR FISH AND SHELLFISH General Provisions Definitions § 60.106 Farm-raised fish. Farm-raised fish means fish or shellfish that have been harvested in controlled environments, including ocean-ranched...

  3. Toward a Theory of Fund Raising in Higher Education.

    ERIC Educational Resources Information Center

    Cook, W. Bruce; Lasher, William F.

    1996-01-01

    A study involving interviews with 50 academic leaders revealed that college fund raising is a team effort; the institution's president is typically the central player; presidents should focus fund-raising attention on major gifts and administrative leadership; academic quality and institutional prestige are critical factors; and fund raising is…

  4. Strategic planning in healthcare organizations.

    PubMed

    Rodríguez Perera, Francisco de Paula; Peiró, Manel

    2012-08-01

    Strategic planning is a completely valid and useful tool for guiding all types of organizations, including healthcare organizations. The organizational level at which the strategic planning process is relevant depends on the unit's size, its complexity, and the differentiation of the service provided. A cardiology department, a hemodynamic unit, or an electrophysiology unit can be an appropriate level, as long as their plans align with other plans at higher levels. The leader of each unit is the person responsible for promoting the planning process, a core and essential part of his or her role. The process of strategic planning is programmable, systematic, rational, and holistic and integrates the short, medium, and long term, allowing the healthcare organization to focus on relevant and lasting transformations for the future.

  5. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    1995-11-15

    The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975 (Public Law 94-163). Its purposes are to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the third quarter of calendar year 1995, including: inventory of petroleum products stored in the Reserve; current storage capacity and ullage available; current status of the Strategic Petroleum Reserve storage facilities, major projects and the acquisition of petroleum products; funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and major environmental actions completed, in progress, or anticipated.

  6. Raising the estimate of functional human sequences.

    PubMed

    Pheasant, Michael; Mattick, John S

    2007-09-01

    While less than 1.5% of the mammalian genome encodes proteins, it is now evident that the vast majority is transcribed, mainly into non-protein-coding RNAs. This raises the question of what fraction of the genome is functional, i.e., composed of sequences that yield functional products, are required for the expression (regulation or processing) of these products, or are required for chromosome replication and maintenance. Many of the observed noncoding transcripts are differentially expressed, and, while most have not yet been studied, increasing numbers are being shown to be functional and/or trafficked to specific subcellular locations, as well as exhibit subtle evidence of selection. On the other hand, analyses of conservation patterns indicate that only approximately 5% (3%-8%) of the human genome is under purifying selection for functions common to mammals. However, these estimates rely on the assumption that reference sequences (usually ancient transposon-derived sequences) have evolved neutrally, which may not be the case, and if so would lead to an underestimate of the fraction of the genome under evolutionary constraint. These analyses also do not detect functional sequences that are evolving rapidly and/or have acquired lineage-specific functions. Indeed, many regulatory sequences and known functional noncoding RNAs, including many microRNAs, are not conserved over significant evolutionary distances, and recent evidence from the ENCODE project suggests that many functional elements show no detectable level of sequence constraint. Thus, it is likely that much more than 5% of the genome encodes functional information, and although the upper bound is unknown, it may be considerably higher than currently thought.

  7. Proglucagon Promoter Cre-Mediated AMPK Deletion in Mice Increases Circulating GLP-1 Levels and Oral Glucose Tolerance

    PubMed Central

    Sayers, Sophie R.; Reimann, Frank; Gribble, Fiona M.; Parker, Helen; Zac-Varghese, Sagen; Bloom, Stephen R.; Foretz, Marc; Viollet, Benoit; Rutter, Guy A.

    2016-01-01

    Background Enteroendocrine L-cells synthesise and release the gut hormone glucagon-like peptide-1 (GLP-1) in response to food transit. Deletion of the tumour suppressor kinase LKB1 from proglucagon-expressing cells leads to the generation of intestinal polyps but no change in circulating GLP-1 levels. Here, we explore the role of the downstream kinase AMP-activated protein kinase (AMPK) in these cells. Method Loss of AMPK from proglucagon-expressing cells was achieved using a preproglucagon promoter-driven Cre (iGluCre) to catalyse recombination of floxed alleles of AMPKα1 and α2. Oral and intraperitoneal glucose tolerance were measured using standard protocols. L-cell mass was measured by immunocytochemistry. Hormone and peptide levels were measured by electrochemical-based luminescence detection or radioimmunoassay. Results Recombination with iGluCre led to efficient deletion of AMPK from intestinal L- and pancreatic alpha-cells. In contrast to mice rendered null for LKB1 using the same strategy, mice deleted for AMPK displayed an increase (WT: 0.05 ± 0.01, KO: 0.09±0.02%, p<0.01) in L-cell mass and elevated plasma fasting (WT: 5.62 ± 0.800 pg/ml, KO: 14.5 ± 1.870, p<0.01) and fed (WT: 15.7 ± 1.48pg/ml, KO: 22.0 ± 6.62, p<0.01) GLP-1 levels. Oral, but not intraperitoneal, glucose tolerance was significantly improved by AMPK deletion, whilst insulin and glucagon levels were unchanged despite an increase in alpha to beta cell ratio (WT: 0.23 ± 0.02, KO: 0.33 ± 0.03, p<0.01). Conclusion AMPK restricts L-cell growth and GLP-1 secretion to suppress glucose tolerance. Targeted inhibition of AMPK in L-cells may thus provide a new therapeutic strategy in some forms of type 2 diabetes. PMID:27010458

  8. Characteristics of Useful and Practical Organizational Strategic Plans

    ERIC Educational Resources Information Center

    Kaufman, Roger

    2014-01-01

    Most organizational strategic plans are not strategic but rather tactical or operational plans masquerading as "strategic." This article identifies the basic elements required in a useful and practical strategic plan and explains why they are important.

  9. Strategic materials availability and supply

    NASA Technical Reports Server (NTRS)

    Stephens, Joseph R.

    1989-01-01

    An evaluation is made of the supply sources and current availability of elements involved in superalloy compositions, as well as of development trends in the substitution of strategic materials for superalloys, giving attention to the results thus far achieved by NASA's Conservation of Strategic Aerospace Materials program, which formally operated only from 1980 to 1984 but whose various research initiatives have been continued under other NASA programs. Peacetime and wartime economies' need and probable availability analyses are presented for bauxite/aluminum, the Pt-group metals, and the alloying elements Mn, Nb, Ta, Co, Cr, Ni, W, Fe, Cu, Ti, Mo, and Mg.

  10. Contrasting strategic and Milan therapies.

    PubMed

    MacKinnon, L

    1983-12-01

    Three related models of therapy are often grouped together as the strategic therapies. These are brief therapy model associated with the Mental Research Institute, approaches developed by Jay Haley and Cloë Madanes, and the model developed by the Milan associates. Controversy exists, however, as to whether the Milan model should be included as a strategic therapy. It appears that the similarities among the three models can mask deeper differences, thus confounding the confusion. This paper contrast the models in their development, theory, and practice.

  11. Final Draft Strategic Marketing Plan.

    SciTech Connect

    United States. Bonneville Power Administration.

    1994-02-01

    The Bonneville Power Administration (BPA) has developed a marketing plan to define how BPA can be viable and competitive in the future, a result important to BPA`s customers and constituents. The Marketing Plan represents the preferred customer outcomes, marketplace achievements, and competitive advantage required to accomplish the Vision and the Strategic Business Objectives of the agency. The Marketing Plan contributes to successful implementation of BPA`s Strategic Business Objectives (SBOs) by providing common guidance to organizations and activities throughout the agency responsible for (1) planning, constructing, operating, and maintaining the Federal Columbia River Power System; (2) conducting business with BPA`s customers; and (3) providing required internal support services.

  12. Putative methyltransferase LaeA and transcription factor CreA are necessary for proper asexual development and controlling secondary metabolic gene cluster expression.

    PubMed

    Zhang, Xiujun; Zhu, Yingying; Bao, Longfei; Gao, Liwei; Yao, Guangshan; Li, Yanan; Yang, Zhifeng; Li, Zhonghai; Zhong, Yaohua; Li, Fuli; Yin, Heng; Qu, Yinbo; Qin, Yuqi

    2016-09-01

    The morphological development of fungi is a complex process and is often coupled with secondary metabolite production. In this study, we assessed the function of putative methyltransferase LaeA and transcription factor CreA in controlling asexual development and secondary metabolic gene cluster expression in Penicillium oxalicum. The deletion of laeA (ΔlaeA) impaired the conidiation in P. oxalicum, with a downregulated expression of brlA. Overexpression of P. oxalicum brlA in ΔlaeA could upregulate brlA and abaA remarkably, but could not rescue the conidiation defect; therefore, brlA and abaA expression were necessary but not sufficient for conidiation. Deletion of creA in ΔlaeA background (ΔlaeAΔcreA) blocked conidiation with a white fluffy phenotype. Nutrient-rich medium could not rescue developmental defects in ΔlaeAΔcreA mutant but could rescue defects in ΔlaeA. Expression of 10 genes, namely, albA/wA, abrB/yA, arpA, aygA, arpA-like, arpB, arpB-like, rodA, rodA-like, and rodB, for pigmentation and spore wall protein genes was silenced in ΔlaeAΔcreA, whereas only six of them were downregulated in ΔlaeA. Among the 28 secondary metabolism gene clusters in P. oxalicum, four secondary metabolism gene clusters were silenced in ΔlaeA and two were also silenced in ΔbrlA mutant. A total of 10 physically linked and coregulated genes were distributed over five chromosomes in ΔlaeA. Six of these genes were located in subtelomeric regions, thus demonstrating a positional bias for LaeA-regulated clusters toward subtelomeric regions. All of silenced clusters located in subtelomeric regions were derepressed in ΔlaeAΔcreA, hence showing that lack of CreA could remediate the repression of gene clusters in ΔlaeA background. Results show that both putative methyltransferase LaeA and transcription factor CreA are necessary for proper asexual development and controlling secondary metabolic gene cluster expression.

  13. Keratin 5-Cre-driven excision of nonmuscle myosin IIA in early embryo trophectoderm leads to placenta defects and embryonic lethality.

    PubMed

    Crish, James; Conti, Mary Anne; Sakai, Takao; Adelstein, Robert S; Egelhoff, Thomas T

    2013-10-01

    In studies initially focused on roles of nonmuscle myosin IIA (NMIIA) in the developing mouse epidermis, we have discovered that a previously described cytokeratin 5 (K5)-Cre gene construct is expressed in early embryo development. Mice carrying floxed alleles of the nonmuscle myosin II heavy chain gene (NMHC IIA(flox/flox)) were crossed with the K5-Cre line. The progeny of newborn pups did not show a Mendelian genotype distribution, suggesting embryonic lethality. Analysis of post-implantation conceptuses from embryonic day (E)9.5 to E13.5 revealed poorly developed embryos and defective placentas, with significantly reduced labyrinth surface area and blood vessel vascularization. These results suggested the novel possibility that the bovine K5 promoter-driven Cre-recombinase was active early in trophoblast-lineage cells that give rise to the placenta. To test this possibility, K5-Cre transgenic mice were crossed with the mT/mG reporter mouse in which activation of GFP expression indicates Cre transgene expression. We observed activation of K5-Cre-driven GFP expression in the ectoplacental cone, in the extraembryonic ectoderm, and in trophoblast giant cells in the E6.5 embryo. In addition, we observed GFP expression at E11.5 to E13.5 in both the labyrinth of the placenta and the yolk sac. NMIIA expression was detected in these same cell types in normal embryos, as well as in E13.5 yolk sac and labyrinth. These findings taken together suggest that NMHC IIA may play critical roles in the early trophoblast-derived ectoplacental cone and extraembryonic ectoderm, as well as in the yolk sac and labyrinth tissues that form later. Our findings are consistent with phenotypes of constitutive NMIIA knockout mice made earlier, that displayed labyrinth and yolk sac-specific defects, but our findings extend those observations by suggesting possible NMIIA roles in trophoblast lineages as well. These results furthermore demonstrate that K5-Cre gene constructs, previously reported

  14. Strategic Planning and Strategic Thinking Clothed in STRATEGO

    ERIC Educational Resources Information Center

    Baaki, John; Moseley, James L.

    2011-01-01

    This article shares experiences that participants had playing the game of STRATEGO and how the activity may be linked to strategic planning and thinking. Among the human performance technology implications of playing this game are that gamers agreed on a framework for rules, took stock on where they wanted to go in the future, and generated a risk…

  15. Strategic Teaching and Strategic Learning in First-Grade Classrooms.

    ERIC Educational Resources Information Center

    Kinnucan-Welsch, Kathryn; Magill, Dodie; Dean, Marie

    1999-01-01

    Describes Reading Express, a collaborative program bringing together first-grade teachers and Reading Recovery teachers to support all first-grade children in literacy development. Indicates that the program is having a positive impact on students' literacy development. Concludes that first-grade teachers are more strategic in their reading…

  16. The Strategic Data Project's Strategic Performance Indicators

    ERIC Educational Resources Information Center

    Page, Lindsay C.; Fullerton, Jon; Bacher-Hicks, Andrew; Owens, Antoniya; Cohodes, Sarah R.; West, Martin R.; Glover, Sarah

    2013-01-01

    Strategic Performance Indicators (SPIs) are summary measures derived from parallel, descriptive analyses conducted across educational agencies. The SPIs are designed to inform agency management and efforts to improve student outcomes. We developed the SPIs to reveal patterns common across partner agencies, to highlight exceptions to those…

  17. Strategic Planning as a Perceptual Process,

    DTIC Science & Technology

    1981-03-01

    Like any complex human endeavor, the sort of broad scope or long range organizational planning often referred to as strategic planning can be viewed...might be of use to planners and managers concerned with broad scope strategic planning .

  18. Strategic Alliance Poker: Demonstrating the Importance of Complementary Resources and Trust in Strategic Alliance Management

    ERIC Educational Resources Information Center

    Reutzel, Christopher R.; Worthington, William J.; Collins, Jamie D.

    2012-01-01

    Strategic Alliance Poker (SAP) provides instructors with an opportunity to integrate the resource based view with their discussion of strategic alliances in undergraduate Strategic Management courses. Specifically, SAP provides Strategic Management instructors with an experiential exercise that can be used to illustrate the value creation…

  19. 76 FR 14950 - Closed Meeting of the U.S. Strategic Command Strategic Advisory Group

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-18

    ... of the Secretary Closed Meeting of the U.S. Strategic Command Strategic Advisory Group AGENCY... notice pertaining to the following federal advisory committee: U.S. Strategic Command Strategic Advisory... meeting is to provide advice on scientific, technical, intelligence, and policy-related issues to...

  20. Strategic Planning and Army Installation Management.

    DTIC Science & Technology

    1996-01-01

    program. The U.S. Army has adopted the Malcolm Baldrige National Quality Award criteria for use in the ACOE program. Strategic planning is one of the...seven pillars of the Baldrige criteria. The Army has recognized that strategic planning is the key to the future. Strategic planning is the key to...and utilization of strategic planning . This paper examines through case study analysis several civilian communities and lessons learned through their

  1. Cell-type specific circuit connectivity of hippocampal CA1 revealed through Cre-dependent rabies tracing

    PubMed Central

    Sun, Yanjun; Nguyen, Amanda; Nguyen, Joseph; Le, Luc; Saur, Dieter; Choi, Jiwon; Callaway, Edward M.; Xu, Xiangmin

    2014-01-01

    Summary We applied a new Cre-dependent, genetically modified rabies-based tracing system to map direct synaptic connections to CA1 excitatory and inhibitory neuron types in mouse hippocampus. We found common inputs to excitatory and inhibitory CA1 neurons from CA3, CA2, entorhinal cortex and the medial septum (MS), and unexpectedly also from the subiculum. Excitatory CA1 neurons receive inputs from both cholinergic and GABAergic MS neurons while inhibitory CA1 neurons receive a great majority of input from GABAergic MS neurons; both cell types also receive weaker input from glutamatergic MS neurons. Comparisons of inputs to CA1 PV+ interneurons versus SOM+ interneurons showed similar strengths of input from the subiculum, but PV+ interneurons receive much stronger input than SOM+ neurons from CA3, entorhinal cortex and MS. Differential input from CA3 to specific CA1 cell types was also demonstrated functionally using laser scanning photostimulation and whole cell recordings. PMID:24656815

  2. Cell-type-specific circuit connectivity of hippocampal CA1 revealed through Cre-dependent rabies tracing.

    PubMed

    Sun, Yanjun; Nguyen, Amanda Q; Nguyen, Joseph P; Le, Luc; Saur, Dieter; Choi, Jiwon; Callaway, Edward M; Xu, Xiangmin

    2014-04-10

    We developed and applied a Cre-dependent, genetically modified rabies-based tracing system to map direct synaptic connections to specific CA1 neuron types in the mouse hippocampus. We found common inputs to excitatory and inhibitory CA1 neurons from CA3, CA2, the entorhinal cortex (EC), the medial septum (MS), and, unexpectedly, the subiculum. Excitatory CA1 neurons receive inputs from both cholinergic and GABAergic MS neurons, whereas inhibitory neurons receive a great majority of inputs from GABAergic MS neurons. Both cell types also receive weaker input from glutamatergic MS neurons. Comparisons of inputs to CA1 PV+ interneurons versus SOM+ interneurons showed similar strengths of input from the subiculum, but PV+ interneurons received much stronger input than SOM+ neurons from CA3, the EC, and the MS. Thus, rabies tracing identifies hippocampal circuit connections and maps how the different input sources to CA1 are distributed with different strengths on each of its constituent cell types.

  3. RecET driven chromosomal gene targeting to generate a RecA deficient Escherichia coli strain for Cre mediated production of minicircle DNA

    PubMed Central

    Tolmachov, Oleg; Palaszewski, Iwona; Bigger, Brian; Coutelle, Charles

    2006-01-01

    Background Minicircle DNA is the non-replicating product of intramolecular site-specific recombination within a bacterial minicircle producer plasmid. Minicircle DNA can be engineered to contain predominantly human sequences which have a low content of CpG dinucleotides and thus reduced immunotoxicity for humans, whilst the immunogenic bacterial origin and antibiotic resistance marker gene sequences are entirely removed by site-specific recombination. This property makes minicircle DNA an excellent vector for non-viral gene therapy. Large-scale production of minicircle DNA requires a bacterial strain expressing tightly controlled site-specific recombinase, such as Cre recombinase. As recombinant plasmids tend to be more stable in RecA-deficient strains, we aimed to construct a recA- bacterial strain for generation of minicircle vector DNA with less chance of unwanted deletions. Results We describe here the construction of the RecA-deficient minicircle DNA producer Escherichia coli HB101Cre with a chromosomally located Cre recombinase gene under the tight control of the araC regulon. The Cre gene expression cassette was inserted into the chromosomal lacZ gene by creating transient homologous recombination proficiency in the recA- strain HB101 using plasmid-born recET genes and homology-mediated chromosomal "pop-in, pop-out" of the plasmid pBAD75Cre containing the Cre gene and a temperature sensitive replication origin. Favourably for the Cre gene placement, at the "pop-out" step, the observed frequency of RecET-led recombination between the proximal regions of homology was 10 times higher than between the distal regions. Using the minicircle producing plasmid pFIXluc containing mutant loxP66 and loxP71 sites, we isolated pure minicircle DNA from the obtained recA- producer strain HB101Cre. The minicircle DNA preparation consisted of monomeric and, unexpectedly, also multimeric minicircle DNA forms, all containing the hybrid loxP66/71 site 5'-TACCGTTCGT ATAATGTATG

  4. A Strategic Plan Is Just the Beginning

    ERIC Educational Resources Information Center

    Wilson, E. B.

    2009-01-01

    The process of strategic planning wears out institutions. Strategic planning, pursued with high purpose and energy, is enervating and exhausting. In this article, the author describes the value-added work of trustees in high-performing boards and discusses the role boards of trustees play as the process of continuous strategic planning unfolds.…

  5. Strategic Activism, Educational Leadership and Social Justice

    ERIC Educational Resources Information Center

    Ryan, James

    2016-01-01

    This article describes the strategic activism of educational leaders who promote social justice. Given the risks, educational leaders need to be strategic about the ways in which they pursue their activism. Citing current research, this article explores the ways in which leaders strategically pursue their social justice agendas within their own…

  6. Promise or Peril: The Strategic Defense Initiative.

    ERIC Educational Resources Information Center

    Brzezinski, Zbigniew, Ed.; And Others

    The major policy debate touched off by President Reagan's March 1983 speech announcing the Strategic Defense Initiative (SDI) was the reopening of one that had begun 35 years before. Then and now the ultimate question is what kind of strategic posture is most likely to contribute to mutual strategic stability? The answer is central to national…

  7. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 13 Business Credit and Assistance 1 2014-01-01 2014-01-01 false Strategic Plans. 313.6 Section 313... § 313.6 Strategic Plans. (a) General. An Impacted Community that intends to apply for a grant for implementation assistance under § 313.7 shall develop and submit a Strategic Plan to EDA for evaluation...

  8. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 13 Business Credit and Assistance 1 2013-01-01 2013-01-01 false Strategic Plans. 313.6 Section 313... § 313.6 Strategic Plans. (a) General. An Impacted Community that intends to apply for a grant for implementation assistance under § 313.7 shall develop and submit a Strategic Plan to EDA for evaluation...

  9. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 13 Business Credit and Assistance 1 2012-01-01 2012-01-01 false Strategic Plans. 313.6 Section 313... § 313.6 Strategic Plans. (a) General. An Impacted Community that intends to apply for a grant for implementation assistance under § 313.7 shall develop and submit a Strategic Plan to EDA for evaluation...

  10. Data and information quality strategic plan.

    PubMed

    Bethell, C

    The Environmental Protection Agency's Strategic Plan was developed in response to internal and external concerns about the integrity, consistency, and accuracy of EPA's environmental data. This document explains why a Strategic Plan is needed and the methodology used in its development, cites Agency models of excellence, and presents the six recommendations of EPA's Data and Information Quality Strategic Plan.

  11. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Strategic plan. 563e.27 Section 563e.27 Banks... for Assessing Performance § 563e.27 Strategic plan. (a) Alternative election. The OTS will assess a... strategic plan if: (1) The savings association has submitted the plan to the OTS as provided for in...

  12. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 3 2010-01-01 2010-01-01 false Strategic plan. 228.27 Section 228.27 Banks and... REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a) Alternative...(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as provided for...

  13. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 3 2013-01-01 2013-01-01 false Strategic plan. 228.27 Section 228.27 Banks and...) COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as...

  14. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 23 Highways 1 2012-04-01 2012-04-01 false Strategic plan. 1335.6 Section 1335.6 Highways NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION STATE HIGHWAY SAFETY DATA IMPROVEMENTS § 1335.6 Strategic plan. A strategic plan shall— (a) Be a multi-year plan that identifies and...

  15. 12 CFR 345.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 5 2012-01-01 2012-01-01 false Strategic plan. 345.27 Section 345.27 Banks and... REINVESTMENT Standards for Assessing Performance § 345.27 Strategic plan. (a) Alternative election. The FDIC... strategic plan if: (1) The bank has submitted the plan to the FDIC as provided for in this section; (2)...

  16. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 6 2014-01-01 2012-01-01 true Strategic plan. 563e.27 Section 563e.27 Banks... for Assessing Performance § 563e.27 Strategic plan. (a) Alternative election. The OTS will assess a... strategic plan if: (1) The savings association has submitted the plan to the OTS as provided for in...

  17. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 3 2014-01-01 2014-01-01 false Strategic plan. 228.27 Section 228.27 Banks and...) COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as...

  18. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 6 2012-01-01 2012-01-01 false Strategic plan. 563e.27 Section 563e.27 Banks... for Assessing Performance § 563e.27 Strategic plan. (a) Alternative election. The OTS will assess a... strategic plan if: (1) The savings association has submitted the plan to the OTS as provided for in...

  19. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 13 Business Credit and Assistance 1 2011-01-01 2011-01-01 false Strategic Plans. 313.6 Section 313... § 313.6 Strategic Plans. (a) General. An Impacted Community that intends to apply for a grant for implementation assistance under § 313.7 shall develop and submit a Strategic Plan to EDA for evaluation...

  20. Collaborative Strategic Planning: Myth or Reality?

    ERIC Educational Resources Information Center

    Mbugua, Flora; Rarieya, Jane F. A.

    2014-01-01

    The concept and practice of strategic planning, while entrenched in educational institutions in the West, is just catching on in Kenya. While literature emphasizes the importance of collaborative strategic planning, it does not indicate the challenges presented by collaboratively engaging in strategic planning. This article reports on findings of…

  1. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    SciTech Connect

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hung, Ming -Szu; Hsieh, David; Au, Alfred; Jablons, David M.; You, Liang

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods: Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (“AdenoCre”) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.

  2. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    DOE PAGES

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; ...

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods:more » Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (“AdenoCre”) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.« less

  3. A 3D Searchable Database of Transgenic Zebrafish Gal4 and Cre Lines for Functional Neuroanatomy Studies.

    PubMed

    Marquart, Gregory D; Tabor, Kathryn M; Brown, Mary; Strykowski, Jennifer L; Varshney, Gaurav K; LaFave, Matthew C; Mueller, Thomas; Burgess, Shawn M; Higashijima, Shin-Ichi; Burgess, Harold A

    2015-01-01

    Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. Using confocal microscopy, we have imaged the cellular-level expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 Gal4 enhancer trap lines, 9 Cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene-specific patterns. Image stacks were acquired at single micron resolution, together with a broadly expressed neural marker, which we used to align enhancer trap reporter patterns into a common 3-dimensional reference space. To facilitate use of this resource, we have written software that enables searching for transgenic lines that label cells within a selectable 3-dimensional region of interest (ROI) or neuroanatomical area. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with co-expression of Cre and Gal4. Many of the imaged enhancer trap lines show intrinsic brain-specific expression. However, to increase the utility of lines that also drive expression in non-neuronal tissue we have designed a novel UAS reporter, that suppresses expression in heart, muscle, and skin through the incorporation of microRNA binding sites in a synthetic 3' untranslated region. Finally, we mapped the site of transgene integration, thus providing molecular identification of the expression pattern for most lines. Cumulatively, this library of enhancer trap lines provides genetic access to 70% of the larval brain and is therefore a powerful and broadly accessible tool for the dissection of neural circuits in larval zebrafish.

  4. A 3D Searchable Database of Transgenic Zebrafish Gal4 and Cre Lines for Functional Neuroanatomy Studies

    PubMed Central

    Marquart, Gregory D.; Tabor, Kathryn M.; Brown, Mary; Strykowski, Jennifer L.; Varshney, Gaurav K.; LaFave, Matthew C.; Mueller, Thomas; Burgess, Shawn M.; Higashijima, Shin-ichi; Burgess, Harold A.

    2015-01-01

    Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. Using confocal microscopy, we have imaged the cellular-level expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 Gal4 enhancer trap lines, 9 Cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene-specific patterns. Image stacks were acquired at single micron resolution, together with a broadly expressed neural marker, which we used to align enhancer trap reporter patterns into a common 3-dimensional reference space. To facilitate use of this resource, we have written software that enables searching for transgenic lines that label cells within a selectable 3-dimensional region of interest (ROI) or neuroanatomical area. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with co-expression of Cre and Gal4. Many of the imaged enhancer trap lines show intrinsic brain-specific expression. However, to increase the utility of lines that also drive expression in non-neuronal tissue we have designed a novel UAS reporter, that suppresses expression in heart, muscle, and skin through the incorporation of microRNA binding sites in a synthetic 3′ untranslated region. Finally, we mapped the site of transgene integration, thus providing molecular identification of the expression pattern for most lines. Cumulatively, this library of enhancer trap lines provides genetic access to 70% of the larval brain and is therefore a powerful and broadly accessible tool for the dissection of neural circuits in larval zebrafish. PMID:26635538

  5. Strategic development: a new focus.

    PubMed

    Lefko, J J

    1989-03-01

    Despite popular convention, a health-care organization's chief executive officer should be the organization's chief strategist. Planners and marketers must begin to function as strategic development officers responsible for sending the right information to the right decision makers so that they can make the right decisions as quickly as possible.

  6. Consultancy on Strategic Information Planning.

    ERIC Educational Resources Information Center

    Pejova, Zdravka, Ed.; Horton, Forest W., Ed.

    At the workshop, better management through strategic planning of information and consultancy was discussed as one way in which developing and Eastern European countries could tackle the complex information problems they are facing during the transition to a market economy. The sixteen papers in this volume are grouped into three basic categories:…

  7. Powerful Metrics: Strategic and Transformative

    ERIC Educational Resources Information Center

    Butterfield, Barbara

    2006-01-01

    To be a valuable partner at the strategic level, human resources can and should contribute to both institutional effectiveness measurement and workforce metrics. In this article, the author examines how to link HR initiatives with key institutional strategies, clarifies essential HR responsibilities for workforce results, explores return on human…

  8. Strategic School Planning in Jordan

    ERIC Educational Resources Information Center

    Al-Zboon, Mohammad Saleem; Hasan, Manal Subhi

    2012-01-01

    The study aimed to measuring the applying degree of the strategic school planning stages at the Governmental high schools from the educational supervisors and principals perspective in the directorates related to Amman city, the study society was formed of the educational supervisors and principals working at Educational directorates related to…

  9. Strategic Communication: A Departmental Transformation

    DTIC Science & Technology

    2010-03-24

    Representatives Adam Smith and Mac Thornberry echoed this view in early March 2010 when they invited other members of the U.S. House of Representatives to...jfq_pages/ editions/i55/1.pdf (accessed January 15, 2010). 5 Ibid. 21 6 U.S. Representatives Adam Smith and Mac Thornberry, “Join the new Strategic

  10. Strategic petroleum reserve annual report

    SciTech Connect

    1996-02-15

    Section 165 of the Energy Policy and Conservation Act (Public Law 94- 163), as amended, requires the Secretary of Energy to submit annual reports to the President and the Congress on activities of the Strategic Petroleum Reserve (SPR). This report describes activities for the year ending December 31, 1995.

  11. Strategic Audit and Marketing Plan

    ERIC Educational Resources Information Center

    Wright, Lianna S.

    2013-01-01

    The purpose of this audit was to revise the marketing plan for ADSum Professional Development School and give the owner a long-term vision of the school to operate competitively in the crowded field of for-profit schools. It is fairly simple to create a strategic plan but harder to implement and execute. Execution requires weeks and months of…

  12. Strategic Planning for Technological Change

    ERIC Educational Resources Information Center

    Strong, Bart

    2007-01-01

    Strategic planning can become downright treacherous if one's institution depends heavily on technology, because technology changes course frequently. Technological change is somewhat predictable, and doing nothing is not an option. A number of complex factors hasten technological change. Limits on performance, breakthrough technology, market…

  13. Strategic Plan for Information Technology.

    ERIC Educational Resources Information Center

    Hudson County Community Coll., Jersey City, NJ.

    This report outlines the Hudson County Community College (HCCC) June 1998 Strategic Plan for Information Technology. After providing a list of planning committee members and an executive summary, the introduction provides background information concerning the creation of the information technology initiative. It also provides the objectives of…

  14. 2002 Industry Studies: Strategic Supply

    DTIC Science & Technology

    2002-01-01

    Strategic Supply is a unique industry study that focused not on a single industry, but rather on a process common to many industries known as Supply ... Chain Management (SCM). We found that the successful, global and innovative companies of today leverage their supply chains to achieve competitive and

  15. Entrepreneurial Spirit in Strategic Planning.

    ERIC Educational Resources Information Center

    Riggs, Donald E.

    1987-01-01

    Presents a model which merges the concepts of entrepreneurship with those of strategic planning to create a library management system. Each step of the process, including needs assessment and policy formation, strategy choice and implementation, and evaluation, is described in detail. (CLB)

  16. NASA Strategic Roadmap Summary Report

    NASA Technical Reports Server (NTRS)

    Wilson, Scott; Bauer, Frank; Stetson, Doug; Robey, Judee; Smith, Eric P.; Capps, Rich; Gould, Dana; Tanner, Mike; Guerra, Lisa; Johnston, Gordon

    2005-01-01

    In response to the Vision, NASA commissioned strategic and capability roadmap teams to develop the pathways for turning the Vision into a reality. The strategic roadmaps were derived from the Vision for Space Exploration and the Aldrich Commission Report dated June 2004. NASA identified 12 strategic areas for roadmapping. The Agency added a thirteenth area on nuclear systems because the topic affects the entire program portfolio. To ensure long-term public visibility and engagement, NASA established a committee for each of the 13 areas. These committees - made up of prominent members of the scientific and aerospace industry communities and senior government personnel - worked under the Federal Advisory Committee Act. A committee was formed for each of the following program areas: 1) Robotic and Human Lunar Exploration; 2) Robotic and Human Exploration of Mars; 3) Solar System Exploration; 4) Search for Earth-Like Planets; 5) Exploration Transportation System; 6) International Space Station; 7) Space Shuttle; 8) Universe Exploration; 9) Earth Science and Applications from Space; 10) Sun-Solar System Connection; 11) Aeronautical Technologies; 12) Education; 13) Nuclear Systems. This document contains roadmap summaries for 10 of these 13 program areas; The International Space Station, Space Shuttle, and Education are excluded. The completed roadmaps for the following committees: Robotic and Human Exploration of Mars; Solar System Exploration; Search for Earth-Like Planets; Universe Exploration; Earth Science and Applications from Space; Sun-Solar System Connection are collected in a separate Strategic Roadmaps volume. This document contains memebership rosters and charters for all 13 committees.

  17. Strategic Planning in Distance Education

    ERIC Educational Resources Information Center

    Kilfoil, Wendy R.

    2003-01-01

    The purpose of strategic planning is to align the institution with the external environment. In fact, it is a complex solution to a complex problem. Environmental change is inevitable and it will always impact one's institution sooner or later. It is better to be proactive, thoughtful, and in charge of change, than to be always in a reactive mode…

  18. Developing a strategic marketing plan.

    PubMed

    Zipin, M L

    1989-06-01

    Strategic planning is essential to the survival of today's hospital. Whether the hospital is an academic medical center, a community hospital or some other type of organization, the key to success is a thorough market planning process. The four-phase process described here focuses on an academic medical center, but it is equally applicable to other types of hospitals.

  19. Strategic Planning to Conduct Joint Force Network Operations: A Content Analysis of NETOPS Organizations Strategic Plans

    DTIC Science & Technology

    2007-03-01

    information dominance , Joint Network Operations (NETOPS) organizations need to be strategically aligned. As result, to enhance the capabilities-based effects of NETOPS and reduce our NETOP infrastructures susceptibility to compromise. Once the key organizations were identified, their strategic plans were analyzed using a structured content analysis framework. The results illustrated that the strategic plans were aligned with the community of interests tasking to conduct NETOPS. Further research is required into the strategic alignment beyond the strategic

  20. Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system.

    PubMed

    Chong-Pérez, Borys; Kosky, Rafael G; Reyes, Maritza; Rojas, Luis; Ocaña, Bárbara; Tejeda, Marisol; Pérez, Blanca; Angenon, Geert

    2012-06-30

    Selectable marker genes are indispensable for efficient production of transgenic events, but are no longer needed after the selection process and may cause public concern and technological problems. Although several gene excision systems exist, few have been optimized for vegetatively propagated crops. Using a Cre-loxP auto-excision strategy, we obtained transgenic banana plants cv. Grande Naine (Musa AAA) devoid of the marker gene used for selection. We used T-DNA vectors with the cre recombinase gene under control of a heat shock promoter and selectable marker gene cassettes placed between two loxP sites in direct orientation, and a gene of interest inserted outside of the loxP sites. Heat shock promoters pGmHSP17.6-L and pHSP18.2, from soybean and Arabidopsis respectively, were tested. A transient heat shock treatment of primary transgenic embryos was sufficient for inducing cre and excising cre and the marker genes. Excision efficiency, as determined by PCR and Southern hybridization was 59.7 and 40.0% for the GmHSP17.6-L and HSP18.2 promoters, respectively. Spontaneous excision was not observed in 50 plants derived from untreated transgenic embryos. To our knowledge this is the first report describing an efficient marker gene removal system for banana. The method described is simple and might be generally applicable for the production of marker-free transgenic plants of many crop species.

  1. Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in L eishmania mexicana cell cycle regulation

    PubMed Central

    Duncan, Samuel M.; Myburgh, Elmarie; Philipon, Cintia; Brown, Elaine; Meissner, Markus; Brewer, James

    2016-01-01

    Summary Leishmania mexicana has a large family of cyclin‐dependent kinases (CDKs) that reflect the complex interplay between cell cycle and life cycle progression. Evidence from previous studies indicated that Cdc2‐related kinase 3 (CRK3) in complex with the cyclin CYC6 is a functional homologue of the major cell cycle regulator CDK1, yet definitive genetic evidence for an essential role in parasite proliferation is lacking. To address this, we have implemented an inducible gene deletion system based on a dimerised Cre recombinase (diCre) to target CRK3 and elucidate its role in the cell cycle of L. mexicana. Induction of diCre activity in promastigotes with rapamycin resulted in efficient deletion of floxed CRK3, resulting in G2/M growth arrest. Co‐expression of a CRK3 transgene during rapamycin‐induced deletion of CRK3 resulted in complementation of growth, whereas expression of an active site CRK3 T178E mutant did not, showing that protein kinase activity is crucial for CRK3 function. Inducible deletion of CRK3 in stationary phase promastigotes resulted in attenuated growth in mice, thereby confirming CRK3 as a useful therapeutic target and diCre as a valuable new tool for analyzing essential genes in Leishmania. PMID:26991545

  2. DMBA/TPA treatment is necessary for BCC formation from patched deficient epidermal cells in Ptch(flox/flox)CD4Cre(+/-) mice.

    PubMed

    Uhmann, Anja; Heß, Ina; Frommhold, Anke; König, Simone; Zabel, Sebastian; Nitzki, Frauke; Dittmann, Kai; Lühder, Fred; Christiansen, Hans; Reifenberger, Julia; Schulz-Schaeffer, Walter; Hahn, Heidi

    2014-10-01

    The development of basal cell carcinoma (BCC), the most frequently diagnosed tumor among persons with European ancestry, is closely linked to mutations in the Hedgehog (Hh) receptor and tumor suppressor Patched1 (Ptch). Using Ptch(flox/flox)CD4Cre(+/-) mice, in which Ptch was ablated in CD4Cre-expressing cells, we demonstrate that the targeted cells can give rise to BCC after treatment with DMBA (7,12-dimethylbenz(a)anthracene)/TPA (12-O-tetradecanoylphorbol-13-acetate), but not after wounding of the skin. In addition, in this model, BCC are not caused by malfunctioning of Ptch-deficient T cells, as BCC did not develop when bone marrow (BM) of Ptch(flox/flox)CD4Cre(+/-) mice was transplanted into Ptch wild-type mice. Instead, lineage-tracing experiments and flow cytometric analyses suggest that the tumors are initiated from rare Ptch-deficient stem cell-like cells of the epidermis that express CD4. As DMBA/TPA is a prerequisite for BCC development in this model, the initiated cells need a second stimulus for expansion and tumor formation. However, in contrast to papilloma, this stimulus seems to be unrelated to alterations in the Ras signaling cascade. Together, these data suggest that biallelic loss of Ptch in CD4(+) cells does not suffice for BCC formation and that BCC formation requires a second so far unknown event, at least in the Ptch(flox/flox)CD4Cre(+/-) BCC mouse model.

  3. BAAV mediated GJB2 gene transfer restores gap junction coupling in cochlear organotypic cultures from deaf Cx26Sox10Cre mice.

    PubMed

    Crispino, Giulia; Di Pasquale, Giovanni; Scimemi, Pietro; Rodriguez, Laura; Galindo Ramirez, Fabian; De Siati, Romolo Daniele; Santarelli, Rosa Maria; Arslan, Edoardo; Bortolozzi, Mario; Chiorini, John A; Mammano, Fabio

    2011-01-01

    The deafness locus DFNB1 contains GJB2, the gene encoding connexin26 and GJB6, encoding connexin30, which appear to be coordinately regulated in the inner ear. In this work, we investigated the expression and function of connexin26 and connexin30 from postnatal day 5 to adult age in double transgenic Cx26(Sox10Cre) mice, which we obtained by crossing connexin26 floxed mice with a deleter Sox10-Cre line. Cx26(Sox10Cre) mice presented with complete connexin26 ablation in the epithelial gap junction network of the cochlea, whereas connexin30 expression was developmentally delayed; immunolabeling patterns for both connexins were normal in the cochlear lateral wall. In vivo electrophysiological measurements in Cx26(Sox10Cre) mice revealed profound hearing loss accompanied by reduction of endocochlear potential, and functional experiments performed in postnatal cochlear organotypic cultures showed impaired gap junction coupling. Transduction of these cultures with a bovine adeno associated virus vector restored connexin26 protein expression and rescued gap junction coupling. These results suggest that restoration of normal connexin levels by gene delivery via recombinant adeno associated virus could be a way to rescue hearing function in DFNB1 mouse models and, in future, lead to the development of therapeutic interventions in humans.

  4. Nucleo-cytoplasmic shuttling dynamics of the transcriptional regulators XYR1 and CRE1 under conditions of cellulase and xylanase gene expression in Trichoderma reesei

    PubMed Central

    Lichius, Alexander; Seidl-Seiboth, Verena; Seiboth, Bernhard; Kubicek, Christian P

    2014-01-01

    Trichoderma reesei is a model for investigating the regulation of (hemi-)cellulase gene expression. Cellulases are formed adaptively, and the transcriptional activator XYR1 and the carbon catabolite repressor CRE1 are main regulators of their expression. We quantified the nucleo-cytoplasmic shuttling dynamics of GFP-fusion proteins of both transcription factors under cellulase and xylanase inducing conditions, and correlated their nuclear presence/absence with transcriptional changes. We also compared their subcellular localization in conidial germlings and mature hyphae. We show that cellulase gene expression requires de novo biosynthesis of XYR1 and its simultaneous nuclear import, whereas carbon catabolite repression is regulated through preformed CRE1 imported from the cytoplasmic pool. Termination of induction immediately stopped cellulase gene transcription and was accompanied by rapid nuclear degradation of XYR1. In contrast, nuclear CRE1 rapidly decreased upon glucose depletion, and became recycled into the cytoplasm. In mature hyphae, nuclei containing activated XYR1 were concentrated in the colony center, indicating that this is the main region of XYR1 synthesis and cellulase transcription. CRE1 was found to be evenly distributed throughout the entire mycelium. Taken together, our data revealed novel aspects of the dynamic shuttling and spatial bias of the major regulator of (hemi-)cellulase gene expression, XYR1, in T. reesei. PMID:25302561

  5. A-MYB (MYBL1) Stimulates Murine Testis-Specific Ldhc Expression via the cAMP-Responsive Element (CRE) Site1

    PubMed Central

    Tang, Huanghui; Goldberg, Erwin

    2011-01-01

    ABSTRACT Generally, knowledge of the mechanism regulating gene expression in primary spermatocytes is incomplete. We have used the lactate dehydrogenase gene (Ldhc) as a model to explore these mechanisms during spermatogenesis. Its 100-bp core promoter contained two essential elements common to many genes, a GC box and a CRE site. Here we report results that support a model in which transcription factor MYBL1 acts as a coactivator directing tissue-specific expression via the CRE cis element. We hypothesize that this is a common mechanism involving activation of multiple genes in the primary spermatocyte. MYBL1 is expressed predominantly as a tissue-specific transcription factor in spermatocytes and breast epithelial cells. Our finding that LDHC expression is lost in 21-day testes of MYBL1 mutant mice supports our hypothesis. In the GC1-spg germ cell line exogenous MYBL1 induces activity 4- to 8-fold, although extracts from these cells do not show MYBL1 binding activity for the Myb consensus sequences in the Ldhc promoter by EMSA. Rather, MYBL1 stimulates expression from a synthetic promoter containing only CRE elements, suggesting MYBL1 activates the promoter by interacting with protein that binds to a CRE element. Mutation of three Myb sites does not affect Ldhc promoter activity significantly (P > 0.05). CREB-binding protein (CBP) is a coactivator that interacts with CRE-binding protein CREB. We show that the transactivation domain (TAD) in MYBL1 interacts with the KIX domain in CBP, and the TAD domain and DNA binding domain in MYBL1 each interact with the CREB N-terminal domain. MYBL1 also stimulated expression from testis-specific genes Pgk2 (phosphoglycerate kinase 2) and Pdha2 (pyruvate dehydrogenase alpha 2) promoters, each of which contains CRE promoter elements and is expressed in primary spermatocytes. We propose that MYBL1 directs germ cell-specific activation via the CRE site of certain genes that are activated specifically in the primary spermatocyte

  6. Daily simulation using a three-dimensional atmosphere-ocean regional coupled model, CReSS-NHOES over the CINDY/DYNAMO observation region

    NASA Astrophysics Data System (ADS)

    Shinoda, T.; Yoshioka, M. K.; Aiki, H.; Kato, M.; Masunaga, H.; Smedstad, L. F.; Katsumata, M.; Yoneyama, K.; Higuchi, A.; Tsuboki, K.; Uyeda, H.

    2012-12-01

    We develop a three-dimensional atmosphere-ocean regional coupled-model with cloud-permitting scale; the atmosphere part is Cloud Resolving Storm Simulator (CReSS) and the ocean one is Non Hydrostatic Ocean model for the Earth Simulator (NHOES). This study shows results of daily simulation over the CINDY/DYNAMO observation region using CReSS-NHOES. Three types of sensitivity experiment are carried out to clarify the effect of the two-way coupled simulation and horizontal grid resolutions. One is the CReSS-NHOES two-way coupled simulation with horizontal grid spacing of 0.045 degree (approximately 4.8 km). Another two simulations are the CReSS simulations without coupling NHOES with horizontal grid spacing of 0.045 and 0.0225 degrees (approximately 2.4 km). The Global Spectral Model (GSM: Horizontal grid resolution is approximately 50 km) data provided by Japan Meteorological Agency (JMA) are used as the initial and boundary conditions of the atmosphere in CReSS and CReSS-NHOES. Three-dimensional Navy Coastal Ocean Model (NCOM) data provided by Naval Research Laboratory are used as the initial and boundary conditions of the ocean in CReSS-NHOES. The daily simulation is carried out for 36 hours from 12 UTC from October 1, 2011 to January 31, 2012 almost every day. We reproduce approximately 30-day surface pressure perturbation that should be related to the Madden-Julian Oscillation, half-day surface pressure perturbation that is related to the atmospheric tide, and the existence of low equivalent potential temperature airmass in the middle troposphere at a fixed observation point of the R/V Mirai (80.5E, 8S). However, the sharp vertical gradient of temperature and salinity at the bottom of the ocean mixed layer at the same point cannot be reproduced. The sensitivity of the coupling of the ocean model is not critical, because the difference of area-averaged sea surface temperature, sensible and latent heat fluxes from the sea surface is quite small. This should be

  7. Endogenous n-3 Polyunsaturated Fatty Acids Delay Progression of Pancreatic Ductal Adenocarcinoma in Fat-1-p48Cre/+-LSL-KrasG12D/+ Mice12

    PubMed Central

    Mohammed, Altaf; Janakiram, Naveena B; Brewer, Misty; Duff, Ashley; Lightfoot, Stan; Brush, Richard S; Anderson, Robert E; Rao, Chinthalapally V

    2012-01-01

    Preclinical studies suggest that diets rich in omega-3 polyunsaturated fatty acids (n-3 PUFAs) may be beneficial for prevention of pancreatic cancer. Nutritional intervention studies are often complex, and there is no clear evidence, without potential confounding factors, on whether conversion of n-6 PUFAs to n-3 PUFAs in pancreatic tissues would provide protection. Experiments were designed using n-3 fatty acid desaturase (Fat-1) transgenic mice, which can convert n-6 PUFA to n-3 FAs endogenously, to determine the impact of n-3 PUFAs on pancreatic intraepithelial neoplasms (PanINs) and their progression to pancreatic ductal adenocarcinoma (PDAC). Six-weekold female p48Cre/+-LSL-KrasG12D/+ andcompoundFat-1-p48Cre/+-LSL-KrasG12D/+ mice were fed (AIN-76A) diets containing 10% safflower oil for 35 weeks. Pancreata were evaluated histopathologically for PanINs and PDAC. Results showed a dramatic reduction in incidence of PDAC (84%; P < .02) in Fat-1-p48Cre/+-LSL-KrasG12D/+ mice compared to p48Cre/+-LSL-KrasG12D/+ mice. Importantly, significant reductions of pancreatic ducts with carcinoma (90%; P < .0001) and PanIN 3 (∼50%; P < .001) lesions were observed in the compound transgenic mice. The levels of n-3 PUFA were much higher (>85%; P < .05–0.01) in pancreas of compound transgenic mice than in those of p48Cre/+-LSL-KrasG12D/+ mice. Molecular analysis of the pancreas showed a significant down-regulation of proliferating cell nuclear antigen, cyclooxygenase-2, 5-lipoxygenase (5-LOX), 5-LOX-activating protein, Bcl-2, and cyclin D1 expression levels in Fat-1-p48Cre/+-LSL-KrasG12D/+ mice compared to p48Cre/+-LSL-KrasG12D/+ mice. These data highlight the promise of dietary n-3 FAs for chemoprevention of pancreatic cancer in high-risk individuals. PMID:23308056

  8. Stress-Induced Anxiety- and Depressive-Like Phenotype Associated with Transient Reduction in Neurogenesis in Adult Nestin-CreERT2/Diphtheria Toxin Fragment A Transgenic Mice.

    PubMed

    Yun, Sanghee; Donovan, Michael H; Ross, Michele N; Richardson, Devon R; Reister, Robin; Farnbauch, Laure A; Fischer, Stephanie J; Riethmacher, Dieter; Gershenfeld, Howard K; Lagace, Diane C; Eisch, Amelia J

    2016-01-01

    Depression and anxiety involve hippocampal dysfunction, but the specific relationship between these mood disorders and adult hippocampal dentate gyrus neurogenesis remains unclear. In both humans with MDD and rodent models of depression, administration of antidepressants increases DG progenitor and granule cell number, yet rodents with induced ablation of DG neurogenesis typically do not demonstrate depressive- or anxiety-like behaviors. The conflicting data may be explained by the varied duration and degree to which adult neurogenesis is reduced in different rodent neurogenesis ablation models. In order to test this hypothesis we examined how a transient-rather than permanent-inducible reduction in neurogenesis would alter depressive- and anxiety-like behaviors. Transgenic Nestin-CreERT2/floxed diphtheria toxin fragment A (DTA) mice (Cre+DTA+) and littermates (Cre+DTA-; control) were given tamoxifen (TAM) to induce recombination and decrease nestin-expressing stem cells and their progeny. The decreased neurogenesis was transient: 12 days post-TAM Cre+DTA+ mice had fewer DG proliferating Ki67+ cells and fewer DCX+ neuroblasts/immature neurons relative to control, but 30 days post-TAM Cre+DTA+ mice had the same DCX+ cell number as control. This ability of DG neurogenesis to recover after partial ablation also correlated with changes in behavior. Relative to control, Cre+DTA+ mice tested between 12-30 days post-TAM displayed indices of a stress-induced anxiety phenotype-longer latency to consume highly palatable food in the unfamiliar cage in the novelty-induced hypophagia test, and a depression phenotype-longer time of immobility in the tail suspension test, but Cre+DTA+ mice tested after 30 days post-TAM did not. These findings suggest a functional association between adult neurogenesis and stress induced anxiety- and depressive-like behaviors, where induced reduction in DCX+ cells at the time of behavioral testing is coupled with stress-induced anxiety and a

  9. Stress-Induced Anxiety- and Depressive-Like Phenotype Associated with Transient Reduction in Neurogenesis in Adult Nestin-CreERT2/Diphtheria Toxin Fragment A Transgenic Mice

    PubMed Central

    Yun, Sanghee; Donovan, Michael H.; Ross, Michele N.; Richardson, Devon R.; Reister, Robin; Farnbauch, Laure A.; Fischer, Stephanie J.; Riethmacher, Dieter; Gershenfeld, Howard K.; Lagace, Diane C.; Eisch, Amelia J.

    2016-01-01

    Depression and anxiety involve hippocampal dysfunction, but the specific relationship between these mood disorders and adult hippocampal dentate gyrus neurogenesis remains unclear. In both humans with MDD and rodent models of depression, administration of antidepressants increases DG progenitor and granule cell number, yet rodents with induced ablation of DG neurogenesis typically do not demonstrate depressive- or anxiety-like behaviors. The conflicting data may be explained by the varied duration and degree to which adult neurogenesis is reduced in different rodent neurogenesis ablation models. In order to test this hypothesis we examined how a transient–rather than permanent–inducible reduction in neurogenesis would alter depressive- and anxiety-like behaviors. Transgenic Nestin-CreERT2/floxed diphtheria toxin fragment A (DTA) mice (Cre+DTA+) and littermates (Cre+DTA-; control) were given tamoxifen (TAM) to induce recombination and decrease nestin-expressing stem cells and their progeny. The decreased neurogenesis was transient: 12 days post-TAM Cre+DTA+ mice had fewer DG proliferating Ki67+ cells and fewer DCX+ neuroblasts/immature neurons relative to control, but 30 days post-TAM Cre+DTA+ mice had the same DCX+ cell number as control. This ability of DG neurogenesis to recover after partial ablation also correlated with changes in behavior. Relative to control, Cre+DTA+ mice tested between 12–30 days post-TAM displayed indices of a stress-induced anxiety phenotype–longer latency to consume highly palatable food in the unfamiliar cage in the novelty-induced hypophagia test, and a depression phenotype–longer time of immobility in the tail suspension test, but Cre+DTA+ mice tested after 30 days post-TAM did not. These findings suggest a functional association between adult neurogenesis and stress induced anxiety- and depressive-like behaviors, where induced reduction in DCX+ cells at the time of behavioral testing is coupled with stress-induced anxiety

  10. Biochemical and mutagenic analysis of I-CreII reveals distinct but important roles for both the H-N-H and GIY-YIG motifs

    PubMed Central

    Corina, Laura E.; Qiu, Weihua; Desai, Ami; Herrin, David L.

    2009-01-01

    Homing endonucleases typically contain one of four conserved catalytic motifs, and other elements that confer tight DNA binding. I-CreII, which catalyzes homing of the Cr.psbA4 intron, is unusual in containing two potential catalytic motifs, H-N-H and GIY-YIG. Previously, we showed that cleavage by I-CreII leaves ends (2-nt 3′ overhangs) that are characteristic of GIY-YIG endonucleases, yet it has a relaxed metal requirement like H-N-H enzymes. Here we show that I-CreII can bind DNA without an added metal ion, and that it binds as a monomer, akin to GIY-YIG enzymes. Moreover, cleavage of supercoiled DNA, and estimates of strand-specific cleavage rates, suggest that I-CreII uses a sequential cleavage mechanism. Alanine substitution of a number of residues in the GIY-YIG motif, however, did not block cleavage activity, although DNA binding was substantially reduced in several variants. Substitution of conserved histidines in the H-N-H motif resulted in variants that did not promote DNA cleavage, but retained high-affinity DNA binding—thus identifying it as the catalytic motif. Unlike the non-specific H-N-H colicins, however; substitution of the conserved asparagine substantially reduced DNA binding (though not the ability to promote cleavage). These results indicate that, in I-CreII, two catalytic motifs have evolved to play important roles in specific DNA binding. The data also indicate that only the H-N-H motif has retained catalytic ability. PMID:19651876

  11. LKB1 deletion with the RIP2.Cre transgene modifies pancreatic β-cell morphology and enhances insulin secretion in vivo

    PubMed Central

    Sun, Gao; Tarasov, Andrei I.; McGinty, James A.; French, Paul M.; McDonald, Angela; Leclerc, Isabelle

    2010-01-01

    The tumor suppressor liver kinase B1 (LKB1), also called STK11, is a protein kinase mutated in Peutz-Jeghers syndrome. LKB1 phosphorylates AMP-activated protein kinase (AMPK) and several related protein kinases. Whereas deletion of both catalytic isoforms of AMPK from the pancreatic β-cell and hypothalamic neurons using the rat insulin promoter (RIP2).Cre transgene (βAMPKdKO) diminishes insulin secretion in vivo, deletion of LKB1 in the β-cell with an inducible Pdx-1.CreER transgene enhances insulin secretion in mice. To determine whether the differences between these models reflect genuinely distinct roles for the two kinases in the β-cell or simply differences in the timing and site(s) of deletion, we have therefore created mice deleted for LKB1 with the RIP2.Cre transgene. In marked contrast to βAMPKdKO mice, βLKB1KO mice showed diminished food intake and weight gain, enhanced insulin secretion, unchanged insulin sensitivity, and improved glucose tolerance. In line with the phenotype of Pdx1-CreER mice, total β-cell mass and the size of individual islets and β-cells were increased and islet architecture was markedly altered in βLKB1KO islets. Signaling by mammalian target of rapamycin (mTOR) to eIF4-binding protein-1 and ribosomal S6 kinase was also enhanced. In contrast to Pdx1-CreER-mediated deletion, the expression of Glut2, glucose-induced changes in membrane potential and intracellular Ca2+ were sharply reduced in βLKB1KO mouse islets and the stimulation of insulin secretion was modestly inhibited. We conclude that LKB1 and AMPK play distinct roles in the control of insulin secretion and that the timing of LKB1 deletion, and/or its loss from extrapancreatic sites, influences the final impact on β-cell function. PMID:20354156

  12. Defining the genome-wide role of CRE1 during carbon catabolite repression in Trichoderma reesei using RNA-Seq analysis.

    PubMed

    Antoniêto, Amanda Cristina Campos; dos Santos Castro, Lílian; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2014-12-01

    The ascomycete Trichoderma reesei is one of the most well-studied cellulolytic fungi and is widely used by the biotechnology industry in the production of second generation bioethanol. The carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1. CCR represses genes related to cellulase production when a carbon source is readily available in the medium. Using RNA sequencing, we investigated CCR during the synthesis of cellulases, comparing the T. reesei Δcre1 mutant strain with its parental strain, QM9414. Of 9129 genes in the T. reesei genome, 268 genes were upregulated and 85 were downregulated in the presence of cellulose (Avicel). In addition, 251 genes were upregulated and 230 were downregulated in the presence of a high concentration of glucose. Genes encoding cellulolytic enzymes and transcription factors and genes related to the transport of nutrients and oxidative metabolism were also targets of CCR, mediated by CRE1 in a carbon source-dependent manner. Our results also suggested that CRE1 regulates the expression of genes related to the use of copper and iron as final electron acceptors or as cofactors of enzymes that participate in biomass degradation. As a result, the final effect of CRE1-mediated transcriptional regulation is to modulate the access of cellulolytic enzymes to cellulose polymers or blocks the entry of cellulase inducers into the cell, depending on the glucose content in the medium. These results will contribute to a better understanding of the mechanism of carbon catabolite repression in T. reesei, thereby enhancing its application in several biotechnology fields.

  13. Inducible gene modification in the gastric epithelium of Tff1-CreERT2, Tff2-rtTA, Tff3-luc mice.

    PubMed

    Thiem, Stefan; Eissmann, Moritz F; Stuart, Emma; Elzer, Joachim; Jonas, Anna; Buchert, Michael; Ernst, Matthias

    2016-12-01

    Temporal and spatial regulation of genes mediated by tissue-specific promoters and conditional gene expression systems provide a powerful tool to study gene function in health, disease, and during development. Although transgenic mice expressing the Cre recombinase in the gastric epithelium have been reported, there is a lack of models that allow inducible and reversible gene modification in the stomach. Here, we exploited the gastrointestinal epithelium-specific expression pattern of the three trefoil factor (Tff) genes and bacterial artificial chromosome transgenesis to generate a novel mouse strain that expresses the CreERT2 recombinase and the reverse tetracycline transactivator (rtTA). The Tg(Tff1-CreERT2;Tff2-rtTA;Tff3-Luc) strain confers tamoxifen-inducible irreversible somatic recombination and allows simultaneous doxycycline-dependent reversible gene activation in the gastric epithelium of developing and adult mice. This strain also confers luciferase activity to the intestinal epithelium to enable in vivo bioluminescence imaging. Using fluorescent reporters as conditional alleles, we show Tff1-CreERT2 and Tff2-rtTA transgene activity in a partially overlapping subset of long-term regenerating gastric stem/progenitor cells. Therefore, the Tg(Tff1-CreERT2;Tff2-rtTA;Tff3-Luc) strain can confer intermittent transgene expression to gastric epithelial cells that have undergone previous gene modification, and may be suitable to genetically model therapeutic intervention during development, tumorigenesis, and other genetically tractable diseases. Birth Defects Research (Part A) 106:626-635, 2016. © 2016 Wiley Periodicals, Inc.

  14. Building a strategic security organisation.

    PubMed

    Howard, Mike

    2016-01-01

    In everyone's day-to-day jobs there is constant need to deal with current and newly detected matters. This is now a world of immediacy, driven by the cadence of the business and its needs. These concerns should not be ignored, as failing to deal with these issues would not bode well for the future. It is essential that the gears are kept spinning. The challenge for any security organisation is to identify its short-term tactical requirements, while developing longer-term strategic needs. Once done, the differences can be accounted for and strides can be made toward a desired future state. This paper highlights several steps that the author and his team have taken in their own journey. There is no magic answer, each organisation will have its own unique challenges. Nevertheless, some of the approaches to building a strategic security organisation described in this paper are applicable to all organisations, irrespective of their size.

  15. Strategic leadership: the essential skills.

    PubMed

    Schoemaker, Paul J H; Krupp, Steve; Howland, Samantha

    2013-01-01

    The more uncertain your environment, the greater the opportunity--if you have the leadership skills to capitalize on it. Research at the Wharton school and at the authors' consulting firm, involving more than 20,000 executives to date, has identified six skills that, when mastered and used in concert, allow leaders to think strategically and navigate the unknown effectively. They are the abilities to anticipate, challenge, interpret, decide, align, and learn. This article describes the six skills in detail and includes a self-assessment that will enable you to identify the ones that most need your attention. The authors have found that strength in one skill cannot easily compensate for a deficit in another. An adaptive strategic leader has learned to apply all six at once.

  16. Strategic Defense Initiative - an overview

    SciTech Connect

    Abrahamson, J.A.

    1985-01-01

    A general framework for developing the concept and implementing options for strategic ballistic missile defense systems is emerging. The objective of the Strategic Defense Initiative (SDI) program is to conduct research on those technologies for defensive system which could intercept ballistic missiles after they have been launched and prevent them from hitting their targets. Its goal is to acquire the technical knowledge as a basis for a later decision on deployment, not a program for deployment or star wars. Abrahamson reviews the basic technological questions facing researchers, outlines the multi-layered defense possibilities and summarizes the status of technology to date. He argues for a continuity of resources to conduct the program. 4 figures.

  17. Strategic Deterrence for the Future

    DTIC Science & Technology

    2015-08-01

    states are investing in their strategic arsenals , developing or modernizing nuclear forces as well as cyber and counterspace capabilities, and thus...States will maintain a safe, secure, and effective arsenal to deter any adversary and to guarantee that defense to our allies.3 In 2011 the “Four...nuclear arsenal . ř In his June 2013 Berlin speech, President Obama announced his assessment that we can ensure the security of America and our allies

  18. Strategic Sourcing in the Army

    DTIC Science & Technology

    2013-09-01

    sector:  Total Cost of Ownership tools are applied to understand the life cycle costs of a product or service  Supplier Scorecards to apply a...sourcing efforts, as well as prioritizing new initiatives. In addition to cost and performance goals, any strategic sourcing plan must be balanced with...article/97687/ 39 INITIAL DISTRIBUTION LIST 1. Defense Technical Information Center Ft. Belvoir, Virginia 2. Dudley Knox Library Naval Postgraduate School Monterey, California

  19. Detention Operations: The Strategic Wildcard

    DTIC Science & Technology

    2011-03-23

    the Master of Strategic Studies Degree. The views expressed in this student academic research paper are those of the author and do not reflect the...time for reviewing instructions , searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this...PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE (DD-MM-YYYY) 23-03-2011 2. REPORT TYPE Strategy Research Project 3. DATES

  20. Case Studies in Strategic Planning

    DTIC Science & Technology

    1990-03-06

    more indepth analysis : "A Theory of Naval Strategic Planning," by LT John R. Hafey, Naval Postgraduate School, Masters Thesis, June 1988, 119 pp. The...research doine on the introduction of competitive strategies (a business concept) into the defense Department. For a more indepth analysis of this...effort, see " Analysis of the Competitive Strategies Methodology (U)," Naval Postgraduate School, by LCDR Michael C. Vitale, USN, Masters’ thesis, December

  1. Strategic Mobility 21 Technical Plan

    DTIC Science & Technology

    2006-09-06

    UNCLASSIFIED Strategic Mobility 21 Technical Plan Contractor Report 0003 Prepared for: Office of Naval Research 875 North Randolph...technical documents or documents containing export-controlled technical data as provided in DoD Directive 5230.25. Standard Form 298 (Rev. 8/98) REPORT ...DOCUMENTATION PAGE Prescribed by ANSI Std. Z39.18 Form Approved OMB No. 0704-0188 The public reporting burden for this collection of information

  2. 2011 Army Strategic Planning Guidance

    DTIC Science & Technology

    2011-03-25

    TESI ) of 22,000 Soldiers, the Army’s total force by the end of the mid-term period is programmed to be 520K (AC). We will achieve a more...dwell ratios, extending TESI authority to adequately man deploying units and sustain the All-Volunteer Force, right-sizing the generating force, and... TESI Temporary End-Strength Increase WMD Weapons of Mass Destruction 2011 ARMY STRATEGIC PLANNING GUIDANCE Page 19 2011

  3. Strategic Vision: A Selected Bibliography

    DTIC Science & Technology

    2008-03-01

    Introduction to Critical Thinking. Amherst: Prometheus Books, 2007. 277pp. (BC177 .C16 2007) Christensen, Ralph. Roadmap to Strategic HR: Turning a...ADA448646 Hall, J. Storrs. Beyond AI: Creating the Conscience of the Machine. Amherst: Prometheus Books, 2007. 408pp. (Q335 .H14 2007) Hall, J. Storrs...Nanofuture: What’s Next for Nanotechnology. Amherst: Prometheus Books, 2005. 333pp. (T174.7 .H14 2005) Hammes, Thomas X. The Sling and the Stone: On War

  4. Strategic petroleum reserve. Quarterly report

    SciTech Connect

    Not Available

    1994-05-15

    The Strategic Petroleum Reserve serves as one of our most important investments in reducing the Nation`s vulnerability to oil supply disruptions. Its existence provides an effective response mechanism should a disruption occur and a formidable deterrent to the use of oil as a political instrument. The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975, (Public Law 94-163) as amended, to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the first quarter of calendar year 1994, including: (1) inventory of petroleum products stored in the Reserve, under contract and in transit at the end of the calendar quarter; (2) fill rate for the current quarter and projected fill rate for the next calendar quarter; (3) average price of the petroleum products acquired during the calendar quarter; (4) current and projected storage capacity; (5) analysis of existing or anticipated problems with the acquisition and storage of petroleum products, and future expansion of storage capacity; (6) funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and (7) major environmental actions completed, in progress, or anticipated.

  5. Strategic petroleum reserve. Quarterly report

    SciTech Connect

    1995-08-15

    The Strategic Petroleum Reserve reduces the Nation`s vulnerability to oil supply disruptions. Its existence provides a formidable deterrent to the use of oil as a political instrument and an effective response mechanism should a disruption occur. The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975 (Public Law 94-163). Its purposes are to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the second quarter of calendar year 1995, including: inventory of petroleum products stored in the Reserve; current and projected storage capacity, analysis of existing or anticipated problems with the acquisition and storage of petroleum products, and future expansion of storage capacity; funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and major environmental actions completed, in progress, or anticipated.

  6. Eye Movements in Strategic Choice

    PubMed Central

    Gächter, Simon; Noguchi, Takao; Mullett, Timothy L.

    2015-01-01

    Abstract In risky and other multiattribute choices, the process of choosing is well described by random walk or drift diffusion models in which evidence is accumulated over time to threshold. In strategic choices, level‐k and cognitive hierarchy models have been offered as accounts of the choice process, in which people simulate the choice processes of their opponents or partners. We recorded the eye movements in 2 × 2 symmetric games including dominance‐solvable games like prisoner's dilemma and asymmetric coordination games like stag hunt and hawk–dove. The evidence was most consistent with the accumulation of payoff differences over time: we found longer duration choices with more fixations when payoffs differences were more finely balanced, an emerging bias to gaze more at the payoffs for the action ultimately chosen, and that a simple count of transitions between payoffs—whether or not the comparison is strategically informative—was strongly associated with the final choice. The accumulator models do account for these strategic choice process measures, but the level‐k and cognitive hierarchy models do not. © 2015 The Authors. Journal of Behavioral Decision Making published by John Wiley & Sons Ltd. PMID:27513881

  7. Grandparents Raising Their Grandchildren: A Comparative Study of Depression

    ERIC Educational Resources Information Center

    Strutton, Joan N.

    2010-01-01

    There has been a steady increase in the United States in recent decades of grandparents raising their grandchildren. The aim of this study was to determine if depression levels of grandparents raising their grandchildren and depression levels of traditional grandparents differ. Additionally, the extent of the relationship to depression scores by…

  8. 12 CFR 1805.503 - Time frame for raising match.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Time frame for raising match. 1805.503 Section 1805.503 Banks and Banking COMMUNITY DEVELOPMENT FINANCIAL INSTITUTIONS FUND, DEPARTMENT OF THE... frame for raising match. Applicants shall satisfy matching funds requirements within the period...

  9. Fund Raising in California School Districts. A Discussion Paper.

    ERIC Educational Resources Information Center

    Allen, Thomas W.; Hughes, K. Scott

    The results of a 1982 survey of California school districts concerning their efforts at private fundraising are presented here. It was found that 123 (or 61 percent) of responding districts either have programs or are planning programs. Although six districts raised more than $100,000 in 1981-82, the majority raised less that $20,000. Of the 66…

  10. HUNTER 10, TRUFLO GONDOLA WEIGHTS AND JACKET. PIVOTING ARMS RAISE ...

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

    HUNTER 10, TRUFLO GONDOLA WEIGHTS AND JACKET. PIVOTING ARMS RAISE THE JACKET AND WEIGHTS. A PNEUMATIC SHOVEL PUSHES THE MORE RECENTLY POURED MOLD INTO THE COOLED MOLD, FORCING THE LATTER ONTO THE VIBRATING SHAKEOUT CONVEYOR AND LEAVING A SPACE UNDER THE RAISED JACKET AND WEIGHTS FOR A FRESHLY MADE UNPOURED MOLD. - Southern Ductile Casting Company, Casting, 2217 Carolina Avenue, Bessemer, Jefferson County, AL

  11. 12 CFR 1805.503 - Time frame for raising match.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 7 2011-01-01 2011-01-01 false Time frame for raising match. 1805.503 Section 1805.503 Banks and Banking COMMUNITY DEVELOPMENT FINANCIAL INSTITUTIONS FUND, DEPARTMENT OF THE... frame for raising match. Applicants shall satisfy matching funds requirements within the period...

  12. 29 CFR 780.615 - Raising of livestock.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Raising of livestock. 780.615 Section 780.615 Labor... Agriculture and Livestock Auction Operations Under the Section 13(b)(13) Exemption Requirements for Exemption § 780.615 Raising of livestock. Livestock auction operations are within the 13(b)(13) exemption...

  13. Constructing an Engineering Model for Raising an Egyptian Obelisk

    ERIC Educational Resources Information Center

    Beck, Charles R.

    2009-01-01

    One of the greatest mysteries of ancient times is how the Egyptians managed to raise huge obelisks using very simple technology. This remarkable task has puzzled engineers for thousand of years. After failing to raise an obelisk with simple machines, such as levers and pulleys, a team of modern engineers solved the mystery using a sandpit and the…

  14. Impact of Education on Grandparents' Actions in Raising Grandchildren

    ERIC Educational Resources Information Center

    Doggett, Diana; Marken, Dory M.; Caldwell, Diana J.

    2014-01-01

    Grandparents raising grandchildren represent a population of adults who confront complex interpersonal and environmental challenges. The intent of this case study was to gather and interpret evaluative data to better understand the impact of a 1-day community education program for grandparents who raise their grandchildren. Extension's philosophy…

  15. Correlates of Fund Raising Outcomes for Four Donor Groups.

    ERIC Educational Resources Information Center

    Loessin, Bruce A.; And Others

    Factors affecting fund raising potential and effectiveness at 575 higher education institutions were investigated, based on fund raising information reported to the Council for Financial Aid to Education for the years 1982-1983, 1983-1984, and 1984-1985. The following types of public and private colleges were studied separately: research,…

  16. Phonological Consciousness Raising Tasks for the ESP Classroom.

    ERIC Educational Resources Information Center

    Sterlacci, Peter

    A discussion of pronunciation instruction in English for business communication focuses on raising learners' awareness of how English is spoken by various language groups around the world. It is argued that phonological consciousness-raising is an effective approach for limiting breakdowns in communication. The approach assumes that developing…

  17. 76 FR 38267 - The Federal Motor Carrier Safety Administration's 2011-2016 Strategic Plan: Raising the Safety Bar

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-29

    ... the specific responsibilities of each party involved in the transport and logistics supply chain in...: Federal eRulemaking Portal: the Federal Docket Management System (FDMS) address: http://www.regulations.gov . Fax: 202-493-2251. Mail: Docket Management Facility (M-30), U.S. Department of...

  18. Implications of the Strategic Defense Initiative for ABM (Anti-Ballistic Missile) Treaty. Professional paper

    SciTech Connect

    Schneiter, G.R.

    1986-02-01

    SDI is a current emphasis of the US strategic program, joining a considerable build-up in US strategic offensive forces. The US defense budget has now begun to reflect increases in research on technologies applicable to a space-based defense. And the United States has enlisted the endorsement of her NATO allies for at least the research portion of SDI. This article examines the effect of this new direction in US defense policy on strategic arms control. It will focus particularly on questions raised for the 1972 Anti-ballistic Missile (ABM) Treaty, in the eyes of many the principal lasting achievement of the era of detente. This Treaty, which bans a territorial defense against strategic ballistic missiles and severely constrains other ABM activities, is the only one of the SALT agreements still legally in force. The article first reviews the strategic forces and programs of the United States and Soviet Union, with emphasis on defensive forces. It then addresses the background and current status of the ABM Treaty. Finally, it identifies and discusses the critical issues with regard to the future of the ABM Treaty.

  19. Enhancement by lithium of cAMP-induced CRE/CREB-directed gene transcription conferred by TORC on the CREB basic leucine zipper domain

    PubMed Central

    Böer, Ulrike; Eglins, Julia; Krause, Doris; Schnell, Susanne; Schöfl, Christof; Knepel, Willhart

    2007-01-01

    The molecular mechanism of the action of lithium salts in the treatment of bipolar disorder is not well understood. As their therapeutic action requires chronic treatment, adaptive neuronal processes are suggested to be involved. The molecular basis of this are changes in gene expression regulated by transcription factors such as CREB (cAMP-response-element-binding protein). CREB contains a transactivation domain, in which Ser119 is phosphorylated upon activation, and a bZip (basic leucine zipper domain). The bZip is involved in CREB dimerization and DNA-binding, but also contributes to CREB transactivation by recruiting the coactivator TORC (transducer of regulated CREB). In the present study, the effect of lithium on CRE (cAMP response element)/CREB-directed gene transcription was investigated. Electrically excitable cells were transfected with CRE/CREB-driven luciferase reporter genes. LiCl (6 mM or higher) induced an up to 4.7-fold increase in 8-bromo-cAMP-stimulated CRE/CREB-directed transcription. This increase was not due to enhanced Ser119 phosphorylation or DNA-binding of CREB. Also, the known targets inositol monophosphatase and GSK3β (glycogen-synthase-kinase 3β) were not involved as specific GSK3β inhibitors and inositol replenishment did not mimic and abolish respectively the effect of lithium. However, lithium no longer enhanced CREB activity when the CREB-bZip was deleted or the TORC-binding site inside the CREB-bZip was specifically mutated (CREB-R300A). Otherwise, TORC overexpression conferred lithium responsiveness on CREB-bZip or the CRE-containing truncated rat somatostatin promoter. This indicates that lithium enhances cAMP-induced CRE/CREB-directed transcription, conferred by TORC on the CREB-bZip. We thus support the hypothesis that lithium salts modulate CRE/CREB-dependent gene transcription and suggest the CREB coactivator TORC as a new molecular target of lithium. PMID:17696880

  20. Securing a biomedical communications future: thinking strategically.

    PubMed

    Stein, D

    1985-11-01

    Ensuring continued growth and viability of the biomedical communication function has become a critical task of the biomedical communications director. Thinking strategically is a cognitive process which assists a director in visualizing programs and tactics which meet clients needs, creates competitive advantages for the biomedical communications unit and builds on existing unit strengths. Thinking strategically can be divided into five phases: strategic vision, strategy development, strategic plan implementation, strategic plan dissemination, and strategic plan evaluation. Each sequence leads the biomedical communications director through a process designed to increase the effectiveness of the biomedical unit and to meet the challenges posed by an environment characterized by diminished financial, material, and human resources as well as respond to threats and opportunities posed by increased competition in the biomedical communications product and marketplace.

  1. Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

    PubMed Central

    Parry, Lee; Young, Madeleine; El Marjou, Fatima; Clarke, Alan Richard

    2015-01-01

    The epithelial surface of the mammalian intestine is a dynamic tissue that renews every 3 - 7 days. Understanding this renewal process identified a population of rapidly cycling intestinal stem cells (ISCs) characterized by their expression of the Lgr5 gene. These are supported by a quiescent stem cell population, marked by Bmi-1 expression, capable of replacing them in the event of injury. Investigating the interactions between these populations is crucial to understanding their roles in disease and cancer. The ISCs exist within crypts on the intestinal surface, these niches support the ISC in replenishing the epithelia. The interaction between active and quiescent ISCs likely involves other differentiated cells within the niche, as it has previously been demonstrated that the ‘‘stemness’’ of the Lgr5 ISC is closely tied to the presence of their neighboring Paneth cells. Using conditional cre-lox mouse models we tested the effect of deleting the majority of active ISCs in the presence or absence of the Paneth cells. Here we describe the techniques and analysis undertaken to characterize the intestine and demonstrate that the Paneth cells play a crucial role within the ISC niche in aiding recovery following substantial insult. PMID:26649885

  2. Textpresso site-specific recombinases: A text-mining server for the recombinase literature including Cre mice and conditional alleles.

    PubMed

    Urbanski, William M; Condie, Brian G

    2009-12-01

    Textpresso Site Specific Recombinases (http://ssrc.genetics.uga.edu/) is a text-mining web server for searching a database of more than 9,000 full-text publications. The papers and abstracts in this database represent a wide range of topics related to site-specific recombinase (SSR) research tools. Included in the database are most of the papers that report the characterization or use of mouse strains that express Cre recombinase as well as papers that describe or analyze mouse lines that carry conditional (floxed) alleles or SSR-activated transgenes/knockins. The database also includes reports describing SSR-based cloning methods such as the Gateway or the Creator systems, papers reporting the development or use of SSR-based tools in systems such as Drosophila, bacteria, parasites, stem cells, yeast, plants, zebrafish, and Xenopus as well as publications that describe the biochemistry, genetics, or molecular structure of the SSRs themselves. Textpresso Site Specific Recombinases is the only comprehensive text-mining resource available for the literature describing the biology and technical applications of SSRs.

  3. Embryonic epithelial Pten deletion through Nkx2.1-cre leads to thyroid tumorigenesis in a strain-dependent manner

    PubMed Central

    Tiozzo, Caterina; Danopoulos, Soula; Lavarreda-Pearce, Maria; Baptista, Sheryl; Varimezova, Radka; Al Alam, Denise; Warburton, David; Virender, Rehan; De Langhe, Stijn; Di Cristofano, Antonio

    2014-01-01

    Even though the role of the tyrosine phosphatase Pten as a tumor suppressor gene has been well established in thyroid cancer, its role during thyroid development is still elusive. We therefore targeted Pten deletion in the thyroid epithelium by crossing Ptenflox/flox with a newly developed Nkx2.1-cre driver line in the BALB/c and C57BL/6 genetic backgrounds. C57BL/6 homozygous Pten mutant mice died around 2 weeks of age due to tracheal and esophageal compression by a hyperplasic thyroid. By contrast, BALB/c homozygous Pten mutant mice survived up to 2 years, but with a slightly increased thyroid volume. Characterization of the thyroid glands from C57BL/6 homozygous Pten mutant mice at postnatal day 14 (PN14) showed abnormally enlarged tissue with areas of cellular hyperplasia, disruption of the normal architecture, and follicular degeneration. In addition, differing degrees of hypothyroidism, thyroxine (T4) decrease, and thyroid-stimulating hormone elevation between the strains in the mutants and the heterozygous mutant were detected at PN14. Finally, C57BL/6 heterozygous Pten mutant mice developed thyroid tumors after 2 years of age. Our results indicate that Pten has a pivotal role in thyroid development and its deletion results in thyroid tumor formation, with the timing and severity of the tumor depending on the particular genetic background. PMID:22167068

  4. Ar-Ar Ages of Nakhlites Y000593, NWA998, and Nakhla and CRE Age of NWA998.

    NASA Technical Reports Server (NTRS)

    Garrison, D. H.; Bogard, D. D.

    2006-01-01

    The seven known Martian nakhlites are Nakhla, Lafayette, Governador Valadares, and four recent finds from hot and cold deserts: MIL03346 from the Transantarctic Mountains, a paired group from the Yamato Mountains (Y000593, Y000749, and Y000802; 1), and two from Morocco (NWA998 and NWA817; 2). Radiometric ages (Sm-Nd, Rb-Sr, U-Pb, and Ar-Ar) for the first three nakhlites, along with Chassigny, fall in the range of 1.19-1.37 Gyr and may suggest a common formation age (e.g., 3). These meteorites also show very similar cosmic-ray (space) exposure ages, which suggests a single ejection event from Mars (3). The ages for nakhlites are different from those of Martian shergottites, whose radiometric ages vary by nearly a factor of three (approximately 165-475 Myr) and whose space exposure ages vary over a factor of approximately 20 (3). Shergottite ages suggest that multiple locations on the Martian surface have been sampled, whereas nakhlite data imply that only one Mars surface location has been sampled. Because older Martian surfaces are expected to be more abundant, it seems surprising that all nakhlites would represent only one Martian impact event. To address this issue, we are measuring the Ar-39-Ar-40 ages of Y-000593, NWA-998, Nakhla, and MIL-03346, and the space (CRE) exposure age of NWA998.

  5. Ar-Ar Ages of Nakhlites Y000593, NWA998, and Nakhla and CRE Ages of NWA998

    NASA Technical Reports Server (NTRS)

    Garrison, D. H.; Bogard, D. D.

    2005-01-01

    The seven known Martian nakhlites are Nakhla, Lafayette, Governador Valadares, and four recent finds from hot and cold deserts: MIL03346 from the Transantarctic Mountains, a paired group from the Yamato Mountains (Y000593, Y000749, and Y000802, and two from Morocco (NWA998 and NWA817. Radiometric ages (Sm-Nd, Rb-Sr, U-Pb, and Ar-Ar) for the first three nakhlites, along with Chassigny, fall in the range of 1.19-1.37 Gyr and may suggest a common formation age. These meteorites also show very similar cosmic-ray (space) exposure ages, suggesting a single ejection event from Mars. The ages for nakhlites are different from those of Martian shergottites, whose radiometric ages vary by nearly a factor of three (approx. 165-475 Myr) and whose space exposure ages vary over a factor of approx. 20. Shergottite ages suggest that multiple locations on the Martian surface have been sampled, whereas nakhlite data imply sampling of only one Mars surface location. Because older Martian surfaces are expected to be more abundant, it seems surprising that all nakhlites would represent only one Martian impact event. To address this issue, we are measuring the (39)Ar-(40)Ar ages of Y-000593, NWA-998, Nakhla, and MIL-03346, and the space (CRE) exposure age of NWA998. Additional information is included in the original extended abstract.

  6. Vaccinia topoisomerase and Cre recombinase catalyze direct ligation of activated DNA substrates containing a 3'-para-nitrophenyl phosphate ester.

    PubMed

    Woodfield, G; Cheng, C; Shuman, S; Burgin, A B

    2000-09-01

    DNA topoisomerases and DNA site-specific recombinases are involved in a diverse set of cellular processes but both function by making transient breaks in DNA. Type IB topoisomerases and tyrosine recombinases cleave DNA by transesterification of an active site tyrosine to generate a DNA-3'-phosphotyrosyl-enzyme adduct and a free 5'-hydroxyl (5'-OH). Strand ligation results when the 5'-OH attacks the covalent complex and displaces the enzyme. We describe the synthesis of 3'-phospho-(para-nitrophenyl) oligonucleotides (3'-pNP DNAs), which mimic the natural 3'-phosphotyrosyl intermediate, and demonstrate that such pre-activated strands are substrates for DNA ligation by vaccinia topoisomerase and Cre recombinase. Ligation occurs by direct attack of a 5'-OH strand on the 3'-pNP DNA (i.e., without a covalent protein-DNA intermediate) and generates free para-nitrophenol as a product. The chromogenic DNA substrate allows ligation to be studied in real-time and in the absence of competing cleavage reactions and can be exploited for high-throughput screening of topoisomerase/recombinase inhibitors.

  7. The measurement of phase equilibria for process streams at operating temperatures and the implications for the CRE liquefaction project

    SciTech Connect

    Waller, A.; Walton, S.T.

    1994-12-31

    CRE operates the Liquid Solvent Extraction process at its pilot plant in North Wales, UK. The process involves feeding pulverised coal, slurried with a solvent, to a digester in which much of the coal is dissolved. Filtration removes residual solids. The coal solution is fed to an ebulliating bed hydrocracker, together with hydrogen at 200 bar at 425{degrees}C. The catalytic reaction adds hydrogen to the coal solution which is distilled to recover solvent and yield a product. The vessels and pipework are externally heated to ensure that liquids can be pumped but temperatures arc not high enough to initiate regressive reactions with consequent formation of carbonaceous solids. However, coal derived liquids are complex mixtures of hydrocarbons which are not soluble in all proportions at all temperatures. Precipitation may occur, leading to blockages in process lines and equipment with subsequent interruption to plant operation. Coal liquids can be regarded simply as a three component system comprising pitch (material with a boiling point >450{degrees}C), saturated hydrocarbon (saturates) having a boiling point <450{degrees}C and distillate solvent, the remainder of the material boiling below 450{degrees}C. Coal solution, of which LSE recycle solvent is a particular case, is a mixture of these three components. A study was undertaken to examine the quantity and nature of precipitates formed from coal solutions of various compositions, with a view to preventing plant shutdowns caused by blocked lines.

  8. The homing endonuclease I-CreI uses three metals, one of which is shared between the two active sites.

    PubMed

    Chevalier, B S; Monnat, R J; Stoddard, B L

    2001-04-01

    Homing endonucleases, like restriction enzymes, cleave double-stranded DNA at specific target sites. The cleavage mechanism(s) utilized by LAGLIDADG endonucleases have been difficult to elucidate; their active sites are divergent, and only one low resolution cocrystal structure has been determined. Here we report two high resolution structures of the dimeric I-CreI homing endonuclease bound to DNA: a substrate complex with calcium and a product complex with magnesium. The bound metals in both complexes are verified by manganese anomalous difference maps. The active sites are positioned close together to facilitate cleavage across the DNA minor groove; each contains one metal ion bound between a conserved aspartate (Asp 20) and a single scissile phosphate. A third metal ion bridges the two active sites. This divalent cation is bound between aspartate residues from the active site of each subunit and is in simultaneous contact with the scissile phosphates of both DNA strands. A metal-bound water molecule acts as the nucleophile and is part of an extensive network of ordered water molecules that are positioned by enzyme side chains. These structures illustrate a unique variant of a two-metal endonuclease mechanism is employed by the highly divergent LAGLIDADG enzyme family.

  9. Is the Army Developing Strategic Leaders?

    DTIC Science & Technology

    2015-06-12

    provide a definition and list of competencies that outlines what a strategic leader should possess. With that definition and list of competencies, the...researcher will examine three military leaders, showing linkage to doctrine, which will illustrate examples of this doctrinal definition . Once this is...Army currently developing strategic leaders? The US Army doctrine and the Strategic Studies Institute (SSI) provide a definition and list of

  10. NERSC Strategic Implementation Plan 2002-2006

    SciTech Connect

    Kramer, William; Bethel, Wes; Craw, James; Draney, Brent; Fortney, William; Gorda, Brend; Harris, William; Meyer, Nancy; Ng, Esmond; Verdier, Francesca; Walter, Howard; Welcome, Tammy

    2002-09-01

    This strategic proposal presents NERSC's vision for its activities and new directions over the next five years. NERSC's continuing commitment to providing high-end systems and comprehensive scientific support for its users will be enhanced, and these activities will be augmented by two new strategic thrusts: support for Scientific Challenge Teams and deployment of a Unified Science Environment. The proposal is in two volumes, the Strategic Plan and the Implementation Plan.

  11. A focused strategic plan for outcomes evaluation.

    PubMed

    Stonestreet, J S; Prevost, S S

    1997-09-01

    This article describes how organizations devise strategic directions. The process of focused strategic or hoshin planning is introduced and reviewed. This model illustrates how a nursing department planned a focused strategic direction for outcomes evaluation. This discussion includes the dedication of resources, the focusing of research priorities and a summary of the outcomes evaluation program, and the method used for implementation of the priorities. Finally, benefits of both the focused planning process and the outcomes evaluation program are discussed.

  12. Hospital strategic preparedness planning: the new imperative.

    PubMed

    Ginter, Peter M; Duncan, W Jack; Abdolrasulnia, Maziar

    2007-01-01

    Strategic preparedness planning is an important new imperative for many hospitals. Strategic preparedness planning goes beyond traditional product/market strategic planning by focusing on disaster prevention, containment, and response roles. Hospitals, because of their unique mission, size, complexity, the types of materials they handle, and the types of patients they encounter, are especially vulnerable to natural and human-initiated disasters. In addition, when disasters occur, hospitals must develop well-conceived first responder (receiver) strategies. This paper argues the case for strategic preparedness planning for hospitals and proposes a process for this relatively new and much needed type of planning.

  13. The cortical activation pattern during bilateral arm raising movements

    PubMed Central

    Jang, Sung Ho; Seo, Jung Pyo; Lee, Seung-Hyun; Jin, Sang-Hyun; Yeo, Sang Seok

    2017-01-01

    Bilateral arm raising movements have been used in brain rehabilitation for a long time. However, no study has been reported on the effect of these movements on the cerebral cortex. In this study, using functional near infrared spectroscopy (fNIRS), we attempted to investigate cortical activation generated during bilateral arm raising movements. Ten normal subjects were recruited for this study. fNIRS was performed using an fNIRS system with 49 channels. Bilateral arm raising movements were performed in sitting position at the rate of 0.5 Hz. We measured values of oxyhemoglobin and total hemoglobin in five regions of interest: the primary sensorimotor cortex, premotor cortex, supplementary motor area, prefrontal cortex, and posterior parietal cortex. During performance of bilateral arm raising movements, oxyhemoglobin and total hemoglobin values in the primary sensorimotor cortex, premotor cortex, supplementary motor area, and prefrontal cortex were similar, but higher in these regions than those in the prefrontal cortex. We observed activation of the arm somatotopic areas of the primary sensorimotor cortex and premotor cortex in both hemispheres during bilateral arm raising movements. According to this result, bilateral arm raising movements appeared to induce large-scale neuronal activation and therefore arm raising movements would be good exercise for recovery of brain functions.

  14. Building Bridges: Fund Raising for Deans, Faculty, and Development Officers.

    ERIC Educational Resources Information Center

    Murphy, Mary Kay, Ed.

    This publication offers 16 articles by higher education administrators, faculty and development officers on dean and faculty members roles in development work as well as the teamwork and synergy that develop in the best efforts. Following an introduction by George Keller, the articles are as follows: (1) "Institutional Strategic Planning:…

  15. A strategic plan to accelerate development of acute stroke treatments.

    PubMed

    Marler, John R

    2012-09-01

    In order to reenergize acute stroke research and accelerate the development of new treatments, we need to transform the usual design and conduct of clinical trials to test for small but significant improvements in effectiveness, and treat patients as soon as possible after stroke onset when treatment effects are most detectable. This requires trials that include thousands of acute stroke patients. A plan to make these trials possible is proposed. There are four components: (1) free access to the electronic medical record; (2) a large stroke emergency network and clinical trial coordinating center connected in real time to hundreds of emergency departments; (3) a clinical trial technology development center; and (4) strategic leadership to raise funds, motivate clinicians to participate, and interact with politicians, insurers, legislators, and other national and international organizations working to advance the quality of stroke care.

  16. Up, Up & Away. Strategic Campaigns.

    ERIC Educational Resources Information Center

    O'Shea, Catherine L.

    1999-01-01

    Uses a ballooning analogy to offer nine suggestions for universities conducting major fund-raising campaigns: study the map and choose a direction; test the prevailing winds; choose and train your crew; gear up for the journey; stay on course; make every bit count; change course as needed; and capitalize on your successful landing. (DB)

  17. A Tet-on and Cre-loxP Based Genetic Engineering System for Convenient Recycling of Selection Markers in Penicillium oxalicum

    PubMed Central

    Jiang, Baojie; Zhang, Ruiqin; Feng, Dan; Wang, Fangzhong; Liu, Kuimei; Jiang, Yi; Niu, Kangle; Yuan, Quanquan; Wang, Mingyu; Wang, Hailong; Zhang, Youming; Fang, Xu

    2016-01-01

    The lack of selective markers has been a key problem preventing multistep genetic engineering in filamentous fungi, particularly for industrial species such as the lignocellulose degrading Penicillium oxalicum JUA10-1(formerly named as Penicillium decumbens). To resolve this problem, we constructed a genetic manipulation system taking advantage of two established genetic systems: the Cre-loxP system and Tet-on system in P. oxalicum JUA10-1. This system is efficient and convenient. The expression of Cre recombinase was activated by doxycycline since it was controlled by Tet-on system. Using this system, two genes, ligD and bglI, were sequentially disrupted by loxP flanked ptrA. The successful application of this procedure will provide a useful tool for genetic engineering in filamentous fungi. This system will also play an important role in improving the productivity of interesting products and minimizing by-product when fermented by filamentous fungi. PMID:27148179

  18. Thrust and parry of the SIOP (single integrated operational plan) and sdi (strategic defense initiative). Research report

    SciTech Connect

    Zank, G.D.

    1989-05-01

    The relationship between strategic offensive capabilities (reflected in the SIOP) and emerging strategic defensive capabilities (reflected by SDI) is not being adequately addressed. A summary of the existing nuclear war planning process is provided, and an analagous defensive process is postulated. Parallels and differences between the two processes are discussed. Potential areas for information exchange and cooperation are identified to enhance deterrence and improve war fighting potential. Operational, technical and political issues requiring resolution are raised and recommendations to resolve these issues are made.

  19. Foxc2(CreERT2) knock-in mice mark stage-specific Foxc2-expressing cells during mouse organogenesis.

    PubMed

    Amin, Mohammed Badrul; Miura, Naoyuki; Uddin, Mohammad Khaja Mafij; Islam, Mohammod Johirul; Yoshida, Nobuaki; Iseki, Sachiko; Kume, Tsutomu; Trainor, Paul A; Saitsu, Hirotomo; Aoto, Kazushi

    2017-01-01

    Foxc2, a member of the winged helix transcription factor family, is essential for eye, calvarial bone, cardiovascular and kidney development in mice. Nevertheless, how Foxc2-expressing cells and their descendent cells contribute to the development of these tissues and organs has not been elucidated. Here, we generated a Foxc2 knock-in (Foxc2(CreERT2) ) mouse, in which administration of estrogen receptor antagonist tamoxifen induces nuclear translocation of Cre recombinase in Foxc2-expressing cells. By crossing with ROSA-LacZ reporter mice (Foxc2(CreERT2) ; R26R), the fate of Foxc2 positive (Foxc2(+) ) cells was analyzed through LacZ staining at various embryonic stages. We found Foxc2(+) cell descendants in the supraoccipital and exoccipital bone in E18.5 embryos, when tamoxifen was administered at embryonic day (E) 8.5. Furthermore, Foxc2(+) descendant cranial neural crest cells at E8-10 were restricted to the corneal mesenchyme, while Foxc2(+) cell derived cardiac neural crest cells at E6-12 were found in the aorta, pulmonary trunk and valves, and endocardial cushions. Foxc2(+) cell descendant contributions to the glomerular podocytes in the kidney were also observed following E6.5 tamoxifen treatment. Our results are consistent with previous reports of Foxc2 expression during early embryogenesis and the Foxc2(CreERT2) mouse provides a tool to investigate spatiotemporal roles of Foxc2 and contributions of Foxc2(+) expressing cells during mouse embryogenesis.

  20. Characterization of Cellulase Secretion and Cre1-Mediated Carbon Source Repression in the Potential Lignocellulose-Degrading Strain Trichoderma asperellum T-1

    PubMed Central

    Wang, Qun; Lin, Hui; Shen, Qi; Fan, Xiaoping; Bai, Naling; Zhao, Yuhua

    2015-01-01

    Trichoderma asperellum, a traditional bio-control species, was demonstrated to be an excellent candidate for lignocellulose degradation in this work. Comparing to the representatively industrial strain of Trichoderma reeseiQM6a, T. asperellum T-1 showed more robust growth, stronger spore production, faster secretion of lignocellulose-decomposing enzymes and better pH tolerance. The reducing sugar released by strain T-1 on the second day of fermentation was 87% higher than that of strain QM6a, although the maximum reducing sugar yield and the cellulase production persistence of the strain T-1 were lower. Our experiment found that the cellulase secretion was strongly inhibited by glucose, suggesting the existence of carbon source repression pathway in T. asperellum T-1. The inhibiting effect was enhanced with an increase in glucose concentration and was closely related to mycelium growth. SDS-PAGE and secondary mass-spectrum identification confirmed that the expression of endo-1,4-β-xylanase I in T. asperellum T-1 was down-regulated when glucose was added. The factor Cre1, which plays an important role in the down-regulation of the endo-1,4-β-xylanase I gene, was investigated by bioinformatics methods. The protein structure of Cre1, analyzed using multiple protein sequence alignment, indicates the existence of the Zn-fingers domain. Then, the binding sites of Cre1 on the endo-1,4-β-xylanase I gene promoter were further elucidated. This study is the first report about Cre1-mediated carbon repression in the bio-control strain T. asperellum T-1. All of the above results provided good references for better understanding T. asperellum T-1 and improving its application for lignocellulose degradation. PMID:25741694