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Sample records for raising cre strategic

  1. 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…

  2. 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fluid lacking pepsin, CD spectroscopy showed that there was a loss of secondary structure and that the protein was no longer active in a functional assay.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.

  3. Raising Student Achievement in the Dayton Public Schools. Report of the Dayton Public Schools Strategic Support Teams.

    ERIC Educational Resources Information Center

    Council of the Great City Schools, Washington, DC.

    This report describes the Dayton Public Schools Strategic Support Teams (SSTs), which address stagnant student achievement among Dayton's students. They are composed of superintendents and other senior urban school managers from across the country who have struggled with the same issues as Dayton. Chapter 1 summarizes SST findings about challenges…

  4. Knockin of Cre Gene at Ins2 Locus Reveals No Cre Activity in Mouse Hypothalamic Neurons

    PubMed Central

    Li, Ling; Gao, Lin; Wang, Kejia; Ma, Xianhua; Chang, Xusheng; Shi, Jian-Hui; Zhang, Ye; Yin, Kai; Liu, Zhimin; Shi, Yuguang; Xie, Zhifang; Zhang, Weiping J.

    2016-01-01

    The recombination efficiency and cell specificity of Cre driver lines are critical for exploring pancreatic β cell biology with the Cre/LoxP approach. Some commonly used Cre lines are based on the short Ins2 promoter fragment and show recombination activity in hypothalamic neurons; however, whether this stems from endogenous Ins2 promoter activity remains controversial. In this study, we generated Ins2-Cre knockin mice with a targeted insertion of IRES-Cre at the Ins2 locus and demonstrated with a cell lineage tracing study that the Ins2 gene is not transcriptionally active in the hypothalamus. The Ins2-Cre driver line displayed robust Cre expression and activity in pancreatic β cells without significant alterations in insulin expression. In the brain, Cre activity was mainly restricted to the choroid plexus, without significant recombination detected in the hippocampus or hypothalamus by the LacZ or fluorescent tdTomato reporters. Furthermore, Ins2-Cre mice exhibited normal glucose tolerance and insulin secretion upon glucose stimulation in vivo. In conclusion, this Ins2-Cre driver line allowed high-fidelity detection of endogenous Ins2 promoter activity in vivo, and the negative activity in the hypothalamus demonstrated that this system is a promising alternative tool for studying β cell biology. PMID:26830324

  5. Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone.

    PubMed

    Oropeza, Daniel; Jouvet, Nathalie; Budry, Lionel; Campbell, Jonathan E; Bouyakdan, Khalil; Lacombe, Julie; Perron, Gabrielle; Bergeron, Valerie; Neuman, Joshua C; Brar, Harpreet K; Fenske, Rachel J; Meunier, Clemence; Sczelecki, Sarah; Kimple, Michelle E; Drucker, Daniel J; Screaton, Robert A; Poitout, Vincent; Ferron, Mathieu; Alquier, Thierry; Estall, Jennifer L

    2015-11-01

    There is growing concern over confounding artifacts associated with β-cell-specific Cre-recombinase transgenic models, raising questions about their general usefulness in research. The inducible β-cell-specific transgenic (MIP-CreERT(1Lphi)) mouse was designed to circumvent many of these issues, and we investigated whether this tool effectively addressed concerns of ectopic expression and disruption of glucose metabolism. Recombinase activity was absent from the central nervous system using a reporter line and high-resolution microscopy. Despite increased pancreatic insulin content, MIP-CreERT mice on a chow diet exhibited normal ambient glycemia, glucose tolerance and insulin sensitivity, and appropriate insulin secretion in response to glucose in vivo and in vitro. However, MIP-CreERT mice on different genetic backgrounds were protected from high-fat/ streptozotocin (STZ)-induced hyperglycemia that was accompanied by increased insulin content and islet density. Ectopic human growth hormone (hGH) was highly expressed in MIP-CreERT islets independent of tamoxifen administration. Circulating insulin levels remained similar to wild-type controls, whereas STZ-associated increases in α-cell number and serum glucagon were significantly blunted in MIP-CreERT(1Lphi) mice, possibly due to paracrine effects of hGH-induced serotonin expression. These studies reveal important new insight into the strengths and limitations of the MIP-CreERT mouse line for β-cell research.

  6. Non-parallel recombination limits Cre-LoxP-based reporters as precise indicators of conditional genetic manipulation.

    PubMed

    Liu, Jing; Willet, Spencer G; Bankaitis, Eric D; Xu, Yanwen; Wright, Chris V E; Gu, Guoqiang

    2013-06-01

    Cre/LoxP-mediated recombination allows for conditional gene activation or inactivation. When combined with an independent lineage-tracing reporter allele, this technique traces the lineage of presumptive genetically modified Cre-expressing cells. Several studies have suggested that floxed alleles have differential sensitivities to Cre-mediated recombination, which raises concerns regarding utilization of Cre-reporters to monitor recombination of other floxed loci of interest. Here, we directly investigate the recombination correlation, at cellular resolution, between several floxed alleles induced by Cre-expressing mouse lines. The recombination correlation between different reporter alleles varied greatly in otherwise genetically identical cell types. The chromosomal location of floxed alleles, distance between LoxP sites, sequences flanking the LoxP sites, and the level of Cre activity per cell all likely contribute to observed variations in recombination correlation. These findings directly demonstrate that, due to non-parallel recombination events, commonly available Cre reporter mice cannot be reliably utilized, in all cases, to trace cells that have DNA recombination in independent-target floxed alleles, and that careful validation of recombination correlations are required for proper interpretation of studies designed to trace the lineage of genetically modified populations, especially in mosaic situations.

  7. 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

  8. 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

    2015-01-01

    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. PMID:26350769

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

    PubMed Central

    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.

    2015-01-01

    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. PMID:26350769

  10. 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.

  11. 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

  12. 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

  13. Perinatal induction of Cre recombination with tamoxifen.

    PubMed

    Lizen, Benoit; Claus, Melissa; Jeannotte, Lucie; Rijli, Filippo M; Gofflot, Françoise

    2015-12-01

    Temporal control of site-specific recombination is commonly achieved by using a tamoxifen-inducible form of Cre or Flp recombinases. Although powerful protocols of induction have been developed for gene inactivation at adult stages or during embryonic development, induction of recombination at late gestational or early postnatal stages is still difficult to achieve. In this context, using the ubiquitous CMV-CreER(T2) transgenic mice, we have tested and validated two procedures to achieve recombination just before and just after birth. The efficiency of recombination was evaluated in the brain, which is known to be more problematic to target. For the late gestation treatment with tamoxifen, different protocols of complementary administration of progesterone and estrogen were tested. However, delayed delivery and/or mortality of pups due to difficult delivery were always observed. To circumvent this problem, pups were collected from tamoxifen-treated pregnant dams by caesarian section at E18.5 and given to foster mothers. For postnatal treatment, different dosages of tamoxifen were administered by intragastric injection to the pups during 3 or 4 days after birth. The efficiency of these treatments was analyzed at P7 using a transgenic reporter line. They were also validated with the Hoxa5 conditional allele. In conclusion, we have developed efficient procedures that allow achieving efficient recombination of floxed alleles at perinatal stages. These protocols will allow investigating the late/adult functions of many developmental genes, whose characterization has been so far restricted to embryonic development. PMID:26395370

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

    PubMed

    Loonstra, A; Vooijs, M; Beverloo, H B; Allak, B A; van Drunen, E; Kanaar, R; Berns, A; Jonkers, J

    2001-07-31

    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-ER(T) 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.

  15. 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…

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

    PubMed

    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.

  17. 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

  18. Cre Activated and Inactivated Recombinant Adeno-Associated Viral Vectors for Neuronal Anatomical Tracing or Activity Manipulation

    PubMed Central

    Saunders, Arpiar

    2015-01-01

    Recombinant adeno-associated viruses (rAAVs) transcriptionally activated by Cre recombinase (Cre-On) are powerful tools for determining the anatomy and function of genetically defined neuronal types in transgenic Cre driver mice. Here we describe how rAAVs transcriptionally inactivated by Cre (Cre-Off) can be used in conjunction with Cre-On rAAVs or genomic Cre-reporter alleles to study brain circuits. Intracranial injection of Cre-On/Cre-Off rAAVs into spatially intermingled Cre+ and Cre- neurons allows these populations to be differentially labeled or manipulated within individual animals. This comparison helps define the unique properties of Cre+ neurons, highlighting the specialized role they play in their constituent brain circuits. This protocol touches on the conceptual and experimental background of Cre-Off rAAV systems, including caveats and methods of validation. PMID:26131660

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

    PubMed

    Vuong, H E; Pérez de Sevilla Müller, L; Hardi, C N; McMahon, D G; Brecha, N C

    2015-10-29

    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) mouse line with three catecholamine-related Cre recombinase mouse 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 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 was 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.

  20. Mosaic pattern of Cre recombinase expression in cochlear outer hair cells of the Brn3.1 Cre mouse.

    PubMed

    Frenz, Silke; Rak, Kristen; Völker, Johannes; Jürgens, Lukas; Scherzad, Agmal; Schendzielorz, Philipp; Radeloff, Andreas; Jablonka, Sibylle; Hansen, Stefan; Mlynski, Robert; Hagen, Rudolf

    2015-04-15

    The Brn3.1 gene encodes for the protein Brn3.1, which is a member of the POU-IV class of transcription factors. Mutation leads to nonsyndromic human progressive hearing loss (DFNA15). To investigate the suitability of the Brn3.1 promoter for Cre recombinase-induced genetic recombination in cochlear hair cells, we established a transgenic Brn3.1 Cre mouse. This mouse line was crossbred with floxed ROSA26 and ROSA26 reporter mice. The cochleae were histologically analysed in cryosections at E16.5 and whole-mount preparations from P2 until P85. In addition, mice from all used strains and their recombinant offspring were tested electrophysiologically by auditory brainstem responses (ABR) and distorsion product otoacoustic emissions (DPOAE). Cre recombinase activity could be detected in P14 and P21 animals in a mosaic pattern in 26.3 and 9.9% of the outer hair cells, respectively. All investigated mice showed normal ABR and DPOAE values, indicating that neither insertion of the internal ribosome entry site (IRES) Cre cassette into the Brn3.1 gene led to abnormal auditory development nor did the reporter strains show inherited hearing disorders. This study shows that Cre expression under the control of the Brn3.1 promoter is feasible and that the insertion of the internal ribosome entry site Cre cassette into this locus exerted no effects on hearing development. Because of the inconstant pattern and the limited duration of expression, the application of the developed mouse line might be restricted. Also, the unchanged hearing capacity and structural integrity of the organ of Corti in available reporter lines indicate that they may be useful tools for hearing research. PMID:25714426

  1. Strategic Leadership

    ERIC Educational Resources Information Center

    Davies, Barbara; Davies, Brent

    2004-01-01

    This article explores the nature of strategic leadership and assesses whether a framework can be established to map the dimensions of strategic leadership. In particular it establishes a model which outlines both the organizational abilities and the individual characteristics of strategic leaders.

  2. 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

  3. Raising Strong Daughters.

    ERIC Educational Resources Information Center

    Gadeberg, Jeanette

    In response to an alarming drop in girls' self-esteem in early adolescence, this parents' guide provides suggestions for raising daughters to become confident, healthy, and independent. Chapter 1, "Yesterday's Daughters," examines how cultural messages inhibit girls' development. Chapter 2, "Raising an Opinionated Daughter," suggests how to help…

  4. Regulation of glycogen metabolism by the CRE-1, RCO-1 and RCM-1 proteins in Neurospora crassa. The role of CRE-1 as the central transcriptional regulator.

    PubMed

    Cupertino, Fernanda Barbosa; Virgilio, Stela; Freitas, Fernanda Zanolli; Candido, Thiago de Souza; Bertolini, Maria Célia

    2015-04-01

    The transcription factor CreA/Mig1/CRE-1 is a repressor protein that regulates the use of alternative carbon sources via a mechanism known as Carbon Catabolite Repression (CCR). In Saccharomyces cerevisiae, Mig1 recruits the complex Ssn6-Tup1, the Neurospora crassa RCM-1 and RCO-1 orthologous proteins, respectively, to bind to promoters of glucose-repressible genes. We have been studying the regulation of glycogen metabolism in N. crassa and the identification of the RCO-1 corepressor as a regulator led us to investigate the regulatory role of CRE-1 in this process. Glycogen content is misregulated in the rco-1(KO), rcm-1(RIP) and cre-1(KO) strains, and the glycogen synthase phosphorylation is decreased in all strains, showing that CRE-1, RCO-1 and RCM-1 proteins are involved in glycogen accumulation and in the regulation of GSN activity by phosphorylation. We also confirmed the regulatory role of CRE-1 in CCR and its nuclear localization under repressing condition in N. crassa. The expression of all glycogenic genes is misregulated in the cre-1(KO) strain, suggesting that CRE-1 also controls glycogen metabolism by regulating gene expression. The existence of a high number of the Aspergillus nidulans CreA motif (5'-SYGGRG-3') in the glycogenic gene promoters led us to analyze the binding of CRE-1 to some DNA motifs both in vitro by DNA gel shift and in vivo by ChIP-qPCR analysis. CRE-1 bound in vivo to all motifs analyzed demonstrating that it down-regulates glycogen metabolism by controlling gene expression and GSN phosphorylation.

  5. 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)

  6. 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)

  7. Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice

    PubMed Central

    Hasegawa, Yoshikazu; Hoshino, Yoshikazu; Ibrahim, Abdelaziz E.; Kato, Kanako; Daitoku, Yoko; Tanimoto, Yoko; Ikeda, Yoshihisa; Oishi, Hisashi; Takahashi, Satoru; Yoshiki, Atsushi; Yagami, Ken-ichi; Iseki, Hiroyoshi; Mizuno, Seiya; Sugiyama, Fumihiro

    2016-01-01

    In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1em1 (cre) Utr strain was produced from an oocyte injected with pX330 containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying 2A-cre. (R26GRR x C57BL/6J-Ins1em1 (cre) Utr) F1 mice were histologically characterized for cre-loxP recombination in the embryonic and adult stages; cre-loxP recombination was observed in all pancreatic islets examined in which almost all insulin-positive cells showed tdsRed fluorescence, suggesting β cell-specific recombination. Furthermore, there were no significant differences in results of glucose tolerance test among genotypes (homo/hetero/wild). Taken together, these observations indicated that C57BL/6J-Ins1em1 (cre) Utr is useful for studies of glucose metabolism and the strategy of bicistronic cre knock-in using the CRISPR/Cas9 system could be useful for production of cre driver mice. PMID:27053096

  8. 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.

  9. Raising Healthy Children.

    ERIC Educational Resources Information Center

    Cummings, Carol; Haggerty, Kevin P.

    1997-01-01

    Notes that teaching social and emotional skills can have a long-term positive effect on academic achievement. The University of Washington's Raising Healthy Children Project tries to reduce children's risk of developing problems in adolescence by providing interventions that bond students to family and school. The program features staff…

  10. SInCRe-structural interactome computational resource for Mycobacterium tuberculosis.

    PubMed

    Metri, Rahul; Hariharaputran, Sridhar; Ramakrishnan, Gayatri; Anand, Praveen; Raghavender, Upadhyayula S; Ochoa-Montaño, Bernardo; Higueruelo, Alicia P; Sowdhamini, Ramanathan; Chandra, Nagasuma R; Blundell, Tom L; Srinivasan, Narayanaswamy

    2015-01-01

    We have developed an integrated database for Mycobacterium tuberculosis H37Rv (Mtb) that collates information on protein sequences, domain assignments, functional annotation and 3D structural information along with protein-protein and protein-small molecule interactions. SInCRe (Structural Interactome Computational Resource) is developed out of CamBan (Cambridge and Bangalore) collaboration. The motivation for development of this database is to provide an integrated platform to allow easily access and interpretation of data and results obtained by all the groups in CamBan in the field of Mtb informatics. In-house algorithms and databases developed independently by various academic groups in CamBan are used to generate Mtb-specific datasets and are integrated in this database to provide a structural dimension to studies on tuberculosis. The SInCRe database readily provides information on identification of functional domains, genome-scale modelling of structures of Mtb proteins and characterization of the small-molecule binding sites within Mtb. The resource also provides structure-based function annotation, information on small-molecule binders including FDA (Food and Drug Administration)-approved drugs, protein-protein interactions (PPIs) and natural compounds that bind to pathogen proteins potentially and result in weakening or elimination of host-pathogen protein-protein interactions. Together they provide prerequisites for identification of off-target binding. PMID:26130660

  11. A system for Cre-regulated RNA interference in vivo

    PubMed Central

    Stern, Patrick; Astrof, Sophie; Erkeland, Stefan J.; Schustak, Joshua; Sharp, Phillip A.; Hynes, Richard O.

    2008-01-01

    We report a system for Cre-regulated expression of RNA interference in vivo. Expression cassettes comprise selectable and FACS-sortable markers in tandem with additional marker genes and shRNAs in the antisense orientation. The cassettes are flanked by tandem LoxP sites arranged so that Cre expression inverts the marker–shRNA construct, allowing its regulated expression (and, at the same time, deletes the original selection/marker genes). The cassettes can be incorporated into retroviral or lentiviral vectors and delivered to cells in culture or used to generate transgenic mice. We describe cassettes incorporating various combinations of reporter genes, miRNA-based RNAi (including two shRNA constructs at once), and oncogenes and demonstrate the delivery of effective RNA interference in cells in culture, efficient transduction into hematopoietic stem cells with cell-type-specific knockdown in their progeny, and rapid generation of regulated shRNA knockdown in transgenic mice. These vector systems allow regulated combinatorial manipulation (both overexpression and loss of function) of gene expression in multiple systems in vitro and in vivo. PMID:18779577

  12. Removal of Carbapenem-Resistant Enterobacteriaceae (CRE) from Blood by Heparin-Functional Hemoperfusion Media

    PubMed Central

    McCrea, Keith; Ward, Robert; LaRosa, Steven P.

    2014-01-01

    Bloodstream infections due to Carbapenem-Resistant Enterobacteriaceae (CRE) are becoming more frequent and are associated with a high mortality. At present, combination antimicrobial therapy yields the best outcomes, but treatment options are limited. Many bacteria utilize heparan sulfate to bind to human cells. We studied the ability of a biomimetic device composed of polyethylene beads with endpoint-attached heparin to bind both sensitive and (CRE) E. coli and Klebsiella pneumoniae from spiked blood samples. Greater than 90% of susceptible, E. coli, CRE E. coli and CRE Klebsiella were removed by the beads. Future studies in human bacteremia with this technology are planned. PMID:25469782

  13. 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.

  14. Pou4f2 knock-in Cre mouse: A multifaceted genetic tool for vision researchers

    PubMed Central

    Simmons, Aaron B.; Bloomsburg, Samuel J.; Billingslea, Samuel A.; Merrill, Morgan M.; Li, Shuai; Thomas, Marshall W.

    2016-01-01

    Purpose: A transgenic mouse that expresses Cre recombinase under control of the Pou4f2-promoter (also referred to as Brn-3b and Brn-3.2) was characterized. Pou4f2 expression has been reported in a subset of retinal ganglion cells (RGCs) in the retina, in the midbrain, and in the germline. In this study, we characterize the expression pattern of this Cre-recombinase line and report its utility in targeted deletion, temporal deletion, RGC depletion, and germline targeting, which can be regulated by the sex of the Cre-carrying mouse. Methods: Pou4f2Cre was mapped by using a combination of PCR and sequencing of PCR products to better understand the construct and to locate where it was inserted within the Pou4f2 locus. Cre expression patterns were examined by crossing Pou4f2Cre/+ mice to Cre reporter mice. Immunohistochemistry was used to further define the pattern of Cre expression and Cre-mediated recombination within the retina, brain, and other tissues. Results: An internal ribosome entry site (IRES)-Cre cassette was inserted into the Pou4f2 gene disrupting normal gene function, as verified by the depletion of RGCs in mice homozygous for the insert. Pou4f2Cre expression was observed in the retina, brain, peripheral neurons, and male germ cells. Germline recombination was observed when the sire carried the Cre and the target for recombination. In all other breeding schemes, recombination was observed within subsets of cells within the retina, brain, intestines, heart, and gonads. In the retina, Cre efficiently targets recombination in neurons within the RGC layer (RGL), the inner nuclear layer (INL), and a small percentage of photoreceptors, activity that has not been previously reported. Unlike most other Cre lines active in the inner retina, recombination in Müller and other glia was not observed in mice carrying Pou4f2Cre. Within the visual centers of the brain, Cre targets recombination in about 15% of cells within the superchiasmatic nucleus, lateral geniculate

  15. The CreC Regulator of Escherichia coli, a New Target for Metabolic Manipulations

    PubMed Central

    Godoy, Manuel S.; Nikel, Pablo I.; Cabrera Gomez, José G.

    2015-01-01

    The CreBC (carbon source-responsive) two-component regulation system of Escherichia coli affects a number of functions, including intermediary carbon catabolism. The impacts of different creC mutations (a ΔcreC mutant and a mutant carrying the constitutive creC510 allele) on bacterial physiology were analyzed in glucose cultures under three oxygen availability conditions. Differences in the amounts of extracellular metabolites produced were observed in the null mutant compared to the wild-type strain and the mutant carrying creC510 and shown to be affected by oxygen availability. The ΔcreC strain secreted more formate, succinate, and acetate but less lactate under low aeration. These metabolic changes were associated with differences in AckA and LdhA activities, both of which were affected by CreC. Measurement of the NAD(P)H/NAD(P)+ ratios showed that the creC510 strain had a more reduced intracellular redox state, while the opposite was observed for the ΔcreC mutant, particularly under intermediate oxygen availability conditions, indicating that CreC affects redox balance. The null mutant formed more succinate than the wild-type strain under both low aeration and no aeration. Overexpression of the genes encoding phosphoenolpyruvate carboxylase from E. coli and a NADH-forming formate dehydrogenase from Candida boidinii in the ΔcreC mutant further increased the yield of succinate on glucose. Interestingly, the elimination of ackA and adhE did not significantly improve the production of succinate. The diverse metabolic effects of this regulator on the central biochemical network of E. coli make it a good candidate for metabolic-engineering manipulations to enhance the formation of bioproducts, such as succinate. PMID:26497466

  16. 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;…

  17. Strategic Bonding.

    ERIC Educational Resources Information Center

    Davis, Lynn; Tyson, Ben

    2003-01-01

    Many school buildings are in dire need of renovation, expansion, or replacement. Brief case studies from around the country illustrate the importance of finding out why people vote for or against a construction referendum. Lists recommendations for a strategic campaign. (MLF)

  18. Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice

    PubMed Central

    Mao, Xiaohong; Fujiwara, Yuko; Orkin, Stuart H.

    1999-01-01

    Effective use of conditional Cre recombinase-loxP gene modification requires Cre-expressing mouse strains with defined patterns of expression. To assess the in vivo functionality of Cre-expressing mice, we have engineered an improved reporter strain for monitoring Cre-mediated excisions. The β-galactosidase-neomycin phosphotransferase fusion gene (βgeo)-trapped ROSA26 locus was modified by gene targeting such that βgeo is expressed only after Cre-mediated excision of loxP-flanked DNA sequences. βgeo from the excised ROSA26 allele is expressed ubiquitously in embryos and adult mice. By mating the reporter strain with Cre-expressing transgenic mice, we have shown that the loxP-flanked ROSA26 allele is accessible to Cre during early embryogenesis, as well as in a specific hematopoietic lineage (T lymphocytes). This improved reporter strain should facilitate monitoring in vivo Cre-mediated excision events in a variety of experimental contexts. PMID:10220414

  19. 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

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

    PubMed

    Felker, A; Mosimann, C

    2016-01-01

    Spatiotemporal transgene regulation by transgenic DNA recombinases is a central tool for reverse genetics in multicellular organisms, with excellent applications for misexpression and lineage tracing experiments. One of the most widespread technologies for this purpose is Cre recombinase-controlled lox site recombination that is attracting increasing interest in the zebrafish field. Tol2-mediated zebrafish transgenesis provides a stable platform to integrate lox cassette transgenes, while the amenability of the zebrafish embryo to drug treatments makes the model an ideal candidate for tamoxifen-inducible CreERT2 experiments. In addition, advanced transgenesis technologies such as phiC31 or CRISPR-Cas9-based knock-ins are even further promoting zebrafish transgenesis for Cre/lox applications. In this chapter, we will first introduce the basics of Cre/lox methodology, CreERT2 regulation by tamoxifen, as well as the utility of Tol2 and other contemporary transgenesis techniques for Cre/lox experiments. We will then outline in detail practical experimental steps for efficient transgenesis toward the creation of single-insertion transgenes and will introduce protocols for 4-hydroxytamoxifen-mediated CreERT2 induction to perform spatiotemporal lox transgene regulation experiments in zebrafish embryos. Last, we will discuss advanced experimental applications of Cre/lox beyond traditional lineage tracing approaches. PMID:27443928

  1. Redesign of the monomer–monomer interface of Cre recombinase yields an obligate heterotetrameric complex

    PubMed Central

    Zhang, Chi; Myers, Connie A.; Qi, Zongtai; Mitra, Robi D.; Corbo, Joseph C.; Havranek, James J.

    2015-01-01

    Cre recombinase catalyzes the cleavage and religation of DNA at loxP sites. The enzyme is a homotetramer in its functional state, and the symmetry of the protein complex enforces a pseudo-palindromic symmetry upon the loxP sequence. The Cre-lox system is a powerful tool for many researchers. However, broader application of the system is limited by the fixed sequence preferences of Cre, which are determined by both the direct DNA contacts and the homotetrameric arrangement of the Cre monomers. As a first step toward achieving recombination at arbitrary asymmetric target sites, we have broken the symmetry of the Cre tetramer assembly. Using a combination of computational and rational protein design, we have engineered an alternative interface between Cre monomers that is functional yet incompatible with the wild-type interface. Wild-type and engineered interface halves can be mixed to create two distinct Cre mutants, neither of which are functional in isolation, but which can form an active heterotetramer when combined. When these distinct mutants possess different DNA specificities, control over complex assembly directly discourages recombination at unwanted half-site combinations, enhancing the specificity of asymmetric site recombination. The engineered Cre mutants exhibit this assembly pattern in a variety of contexts, including mammalian cells. PMID:26365240

  2. 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

  3. Pdgfrb‐Cre targets lymphatic endothelial cells of both venous and non‐venous origins

    PubMed Central

    Ulvmar, Maria H.; Martinez‐Corral, Ines; Stanczuk, Lukas

    2016-01-01

    Abstract The Pdgfrb‐Cre line has been used as a tool to specifically target pericytes and vascular smooth muscle cells. Recent studies showed additional targeting of cardiac and mesenteric lymphatic endothelial cells (LECs) by the Pdgfrb‐Cre transgene. In the heart, this was suggested to provide evidence for a previously unknown nonvenous source of LECs originating from yolk sac (YS) hemogenic endothelium (HemEC). Here we show that Pdgfrb‐Cre does not, however, target YS HemEC or YS‐derived erythro‐myeloid progenitors (EMPs). Instead, a high proportion of ECs in embryonic blood vessels of multiple organs, as well as venous‐derived LECs were targeted. Assessment of temporal Cre activity using the R26‐mTmG double reporter suggested recent occurrence of Pdgfrb‐Cre recombination in both blood and lymphatic ECs. It thus cannot be excluded that Pdgfrb‐Cre mediated targeting of LECs is due to de novo expression of the Pdgfrb‐Cre transgene or their previously established venous endothelial origin. Importantly, Pdgfrb‐Cre targeting of LECs does not provide evidence for YS HemEC origin of the lymphatic vasculature. Our results highlight the need for careful interpretation of lineage tracing using constitutive Cre lines that cannot discriminate active from historical expression. The early vascular targeting by the Pdgfrb‐Cre also warrants consideration for its use in studies of mural cells. genesis 54:350–358, 2016. © 2016 The Authors. Genesis Published by Wiley Periodicals, Inc. PMID:27060598

  4. 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. PMID:26083735

  5. [Deletion of marker gene in transgenic goat by Cre/LoxP system].

    PubMed

    Lan, Chong; Ren, Lina; Wu, Min; Liu, Siguo; Liu, Guohui; Xu, Xujun; Chen, Jianquan; Ma, Hengdong; Cheng, Guoxiang

    2013-12-01

    In producing transgenic livestock, selectable marker genes (SMGs) are usually used to screen transgenic cells from numerous normal cells. That results in SMGs integrating into the genome and transmitting to offspring. In fact, SMGs could dramatically affect gene regulation at integration sites and also make the safety evaluation of transgenic animals complicated. In order to determine the deletion time and methods in the process of producing transgenic goats, the feasibility of deleting SMGs was explored by Cre/LoxP before or after somatic cell cloning. In addition, we compared the efficiency of protein transduction with plasmids co-transduction. We could delete 43.9% SMGs after screening out the transgenic cell clones, but these cells could not be applied to somatic cells cloning because of serious aging after two gene modifications. The SMG-free cells suitable for nuclear transfer were accessible by using the cells of transgenic goats, but this approach was more time consuming. Finally, we found that the Cre plasmid could delete SMGs with an efficiency of 7.81%, but about 30% in SMG-free cells had sequences of Cre plasmid. Compared with Cre plasmid, the integration of new exogenous gene could be avoided by TAT-CRE protein transduction, and the deletion rate of TAT-CRE transduction was between 43.9 and 72.8%. Therefore, TAT-Cre transduction could be an effective method for deleting selectable marker genes.

  6. 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.

  7. 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. PMID:21924163

  8. 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.

  9. 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

  10. Nociceptor-specific gene deletion using heterozygous NaV1.8-Cre recombinase mice.

    PubMed

    Stirling, L Caroline; Forlani, Greta; Baker, Mark D; Wood, John N; Matthews, Elizabeth A; Dickenson, Anthony H; Nassar, Mohammed A

    2005-01-01

    NaV1.8 is a voltage-gated sodium channel expressed only in a subset of sensory neurons of which more than 85% are nociceptors. In order to delete genes in nociceptive neurons, we generated heterozygous transgenic mice expressing Cre recombinase under the control of the NaV1.8 promoter. Functional Cre recombinase expression replicated precisely the expression pattern of NaV1.8. Cre expression began at embryonic day 14 in small diameter neurons in dorsal root, trigeminal and nodose ganglia, but was absent in non-neuronal or CNS tissues into adulthood. Sodium channel subtypes were normal in isolated DRG neurons. Pain behaviour in response to mechanical or thermal stimuli, and in acute, inflammatory and neuropathic pain was also normal. These data demonstrate that the heterozygous NaV1.8-Cre mouse line is a useful tool to analyse the effects of deleting floxed genes on pain behaviour. PMID:15621361

  11. 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 ...

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

    PubMed Central

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

    2015-01-01

    Summary 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 (Tnnt2MerCreMer/+) 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 Tnnt2MerCreMer/+ mouse could be applied towards the temporal genetic deletion of genes of interests in cardiomyocytes with Cre-LoxP technology. The Tnnt2MerCreMer/+ mouse model also provides a useful tool to trace myocardial lineage during development and repair after cardiac injury. PMID:26010701

  13. FlEx-based transgenic reporter lines for visualization of Cre and Flp activity in live zebrafish

    PubMed Central

    Boniface, Emily J.; Lu, Jianjun; Vicotroff, Tristan; Zhu, Meiying; Chen, Wenbiao

    2013-01-01

    Summary Site-specific recombinases such as Cre and Flp are invaluable tools for genetic manipulations, but their usage in zebrafish has been limited. Incorporating recently developed flip-excision (FlEx) design that allows stable inversions, we have established zebrafish reporter lines that express bright and ubiquitous EGFP, but switch to express mCherry in the presence of Cre or Flp. Here, we demonstrate the stable inversion in the reporter lines, both in somatic cells and in the germ line by Cre or Flp, and the subsequent reinversion using the other recombinase. Using the reporter lines, we characterized cardiomyocyte-specific Cre lines and neuronal progenitor-specific and tamoxifen-dependent Cre lines. We also used the reporter lines for screening Cre- and Flp-based enhancer trap lines. Similar to the widely used Cre reporter lines in mice, these FlEx-based reporter lines will facilitate the use of recombinases for genetic manipulations in zebrafish. PMID:19415631

  14. FlEx-based transgenic reporter lines for visualization of Cre and Flp activity in live zebrafish.

    PubMed

    Boniface, Emily J; Lu, Jianjun; Victoroff, Tristan; Zhu, Meiying; Chen, Wenbiao

    2009-07-01

    Site-specific recombinases such as Cre and Flp are invaluable tools for genetic manipulations, but their usage in zebrafish has been limited. Incorporating recently developed flip-excision (FlEx) design that allows stable inversions, we have established zebrafish reporter lines that express bright and ubiquitous EGFP, but switch to express mCherry in the presence of Cre or Flp. Here, we demonstrate the stable inversion in the reporter lines, both in somatic cells and in the germ line by Cre or Flp, and the subsequent reinversion using the other recombinase. Using the reporter lines, we characterized cardiomyocyte-specific Cre lines and neuronal progenitor-specific and tamoxifen-dependent Cre lines. We also used the reporter lines for screening Cre- and Flp-based enhancer trap lines. Similar to the widely used Cre reporter lines in mice, these FlEx-based reporter lines will facilitate the use of recombinases for genetic manipulations in zebrafish.

  15. Screening strategy to generate cell specific recombination: a case report with the RIP-Cre mice.

    PubMed

    Spinelli, Valeria; Martin, Céline; Dorchies, Emilie; Vallez, Emmanuelle; Dehondt, Hélène; Trabelsi, Mohamed-Sami; Tailleux, Anne; Caron, Sandrine; Staels, Bart

    2015-10-01

    Conditional gene knockout technology is a powerful tool to study the function of a gene in a specific tissue, organ or cell lineage. The most commonly used procedure applies the Cre-LoxP strategy, where the choice of the Cre driver promoter is critical to determine the efficiency and specificity of the system. However, a considered choice of an appropriate promoter does not always protect against the risk of unwanted recombination and the consequent deletion of the gene in other tissues than the desired one(s), due to phenomena of non-specific activation of the Cre transgene. Furthermore, the causes of these phenomena are not completely understood and this can potentially affect every strain of Cre-mice. In our study on the deletion of a same gene in two different tissues, we show that the incidence rate of non-specific recombination in unwanted tissues depends on the Cre driver strain, ranging from 100%, rendering it useless (aP2-Cre strain), to ~5%, which is still compatible with their use (RIP-Cre strain). The use of a simple PCR strategy conceived to detect this occurrence is indispensable when producing a tissue-specific knockout mouse. Therefore, when choosing the Cre-driver promoter, researchers not only have to be careful about its tissue-specificity and timing of activation, but should also include a systematical screening in order to exclude mice in which atypical recombination has occurred and to limit the unnecessary use of laboratory animals in uninterpretable experiments. PMID:26091792

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

    PubMed

    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

  17. 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

  18. Behavioral phenotyping of Nestin-Cre mice: implications for genetic mouse models of psychiatric disorders.

    PubMed

    Giusti, Sebastian A; Vercelli, Claudia A; Vogl, Annette M; Kolarz, Adam W; Pino, Natalia S; Deussing, Jan M; Refojo, Damian

    2014-08-01

    Genetic mouse models based on the Cre-loxP system have been extensively used to explore the influence of specific gene deletions on different aspects of behavioral neurobiology. However, the interpretation of the effects attributed to the gene deletion might be obscured by potential side effects secondary to the Cre recombinase transgene insertion or Cre activity, usually neither controlled nor reported. Here, we performed a comprehensive behavioral analysis of endophenotypes of neuropsychiatric disorders in the extensively used Nestin(Cre) mouse line, commonly employed to restrict genetic modifications to the CNS. We observed no alterations in locomotion, general exploratory activity, learning and memory, sociability, startle response and sensorimotor gating. Although the overall response to stimuli triggering anxiety-like behaviors remained unaltered in Nestin(Cre) mice, a strong impairment in the acquisition of both contextual- and cued-conditioned fear was observed. These results underline the importance of adequately controlling the behavioral performance of the employed Cre-lines per-se in pre-clinical neurobehavioral research.

  19. Spatially-Directed Assembly of a Heterotetrameric Cre-Lox Synapse Restricts Recombination Specificity

    PubMed Central

    Gelato, Kathy A.; Martin, Shelly S.; Liu, Patty H.; Saunders, April A.; Baldwin, Enoch P.

    2008-01-01

    SUMMARY The pseudo-fourfold homo-tetrameric synapse formed by Cre protein and target DNA restricts site-specific recombination to sequences containing dyad-symmetric Cre-binding repeats. Mixtures of engineered altered-specificity Cre monomers can form heterotetramers that recombine non-identical asymmetric sequences, allowing greater flexibility for target site selection in the genome of interest. However, the variety of tetramers allowed by random subunit association increases the chances of unintended reactivity at non-target sites. This problem can be circumvented by specifying a unique spatial arrangement of heterotetramer subunits. By reconfiguring inter-subunit protein-protein contacts, we directed the assembly of two different Cre monomers, each having a distinct DNA sequence specificity, in an alternating (ABAB) configuration. This designed heterotetramer preferentially recombined a particular pair of asymmetric Lox sites over other pairs, whereas a mixture of freely-associating subunits showed little bias. Alone, the engineered monomers had reduced reactivity towards both dyad-symmetric and asymmetric sites. Specificity arose because the organization of Cre-binding repeats of the preferred substrate matched the programmed arrangement of the subunits in the heterotetrameric synapse. Applying this “spatial matching” principle, Cre-mediated recombination can be directed to asymmetric DNA sequences with greater fidelity. PMID:18374357

  20. 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.

  1. 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

  2. A New Strategic Planning Model for Academic Libraries.

    ERIC Educational Resources Information Center

    Birdsall, Douglas G.; Hensley, Oliver D.

    1994-01-01

    Presents a strategic planning model originally developed for university research and illustrates its usefulness for academic libraries. Library development and fund-raising activities are used as examples to illustrate the various stages of strategic planning, including scanning the environment, designing the plan, acceptance, and adoption.…

  3. 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…

  4. Construction of Marker-Free Transgenic Strains of Chlamydomonas reinhardtii Using a Cre/loxP-Mediated Recombinase System.

    PubMed

    Kasai, Yuki; Harayama, Shigeaki

    2016-01-01

    The Escherichia coli bacteriophage P1 encodes a site-specific recombinase called Cre and two 34-bp target sites of Cre recombinase called loxP. The Cre/loxP system has been used to achieve targeted insertion and precise deletion in many animal and plant genomes. The Cre/loxP system has particularly been used for the removal of selectable marker genes to create marker-free transgenic organisms. For the first time, we applied the Cre/loxP-mediated site-specific recombination system to Chlamydomonas reinhardtii to construct marker-free transgenic strains. Specifically, C. reinhardtii strains cc4350 and cc124 carrying an aphVIII expression cassette flanked by two direct repeats of loxP were constructed. Separately, a synthetic Cre recombinase gene (CrCRE), the codons of which were optimized for expression in C. reinhardtii, was synthesized, and a CrCRE expression cassette was introduced into strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette. Among 46 transformants carrying the CrCRE expression cassette stably, the excision of aphVIII by CrCre recombinase was observed only in one transformant. We then constructed an expression cassette of an in-frame fusion of ble to CrCRE via a short linker peptide. The product of ble (Ble) is a bleomycin-binding protein that confers resistance to bleomycin-related antibiotics such as Zeocin and localizes in the nucleus. Therefore, the ble-(linker)-CrCRE fusion protein is expected to localize in the nucleus. When the ble-(linker)-CrCRE expression cassette was integrated into the genome of strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette, CrCre recombinase-mediated excision of the aphVIII expression cassette was observed at a frequency higher than that in stable transformants of the CrCRE expression cassette. Similarly, from strain cc124 carrying a single loxP-flanked aphVIII expression cassette, the aphVIII expression cassette was successfully excised after

  5. Construction of Marker-Free Transgenic Strains of Chlamydomonas reinhardtii Using a Cre/loxP-Mediated Recombinase System

    PubMed Central

    Kasai, Yuki; Harayama, Shigeaki

    2016-01-01

    The Escherichia coli bacteriophage P1 encodes a site-specific recombinase called Cre and two 34-bp target sites of Cre recombinase called loxP. The Cre/loxP system has been used to achieve targeted insertion and precise deletion in many animal and plant genomes. The Cre/loxP system has particularly been used for the removal of selectable marker genes to create marker-free transgenic organisms. For the first time, we applied the Cre/loxP-mediated site-specific recombination system to Chlamydomonas reinhardtii to construct marker-free transgenic strains. Specifically, C. reinhardtii strains cc4350 and cc124 carrying an aphVIII expression cassette flanked by two direct repeats of loxP were constructed. Separately, a synthetic Cre recombinase gene (CrCRE), the codons of which were optimized for expression in C. reinhardtii, was synthesized, and a CrCRE expression cassette was introduced into strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette. Among 46 transformants carrying the CrCRE expression cassette stably, the excision of aphVIII by CrCre recombinase was observed only in one transformant. We then constructed an expression cassette of an in-frame fusion of ble to CrCRE via a short linker peptide. The product of ble (Ble) is a bleomycin-binding protein that confers resistance to bleomycin-related antibiotics such as Zeocin and localizes in the nucleus. Therefore, the ble-(linker)-CrCRE fusion protein is expected to localize in the nucleus. When the ble-(linker)-CrCRE expression cassette was integrated into the genome of strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette, CrCre recombinase-mediated excision of the aphVIII expression cassette was observed at a frequency higher than that in stable transformants of the CrCRE expression cassette. Similarly, from strain cc124 carrying a single loxP-flanked aphVIII expression cassette, the aphVIII expression cassette was successfully excised after

  6. 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)

  7. 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

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

    PubMed

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

    2016-01-01

    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 spatiotemporal 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.

  9. 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. PMID:27077909

  10. 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.

  11. Tamoxifen administration routes and dosage for inducible Cre-mediated gene disruption in mouse hearts.

    PubMed

    Andersson, Kristin B; Winer, Lisbeth H; Mørk, Halvor K; Molkentin, Jeffery D; Jaisser, Frédéric

    2010-08-01

    Tissue-specific and time-dependent control of in vivo gene disruption may be achieved using conditional knockout strategies in transgenic mice. Fusion of mutant estrogen receptor ligand-binding domains to Cre recombinase (Cre-ER(T), MerCreMer) combined with cardiac-directed gene expression has been used to generate several cardiac-specific tamoxifen-inducible Cre-expressing mouse lines. Such mice have successfully been used to generate Cre-loxP-mediated gene disruption in an inducible manner in the myocardium in vivo. However, information is sparse regarding the tamoxifen dosage, the time course of gene disruption and whether different administration routes differ in efficiency in obtaining gene disruption in the myocardium. We have evaluated these parameters in Serca2 ( flox/flox ) Tg(alphaMHC-MerCreMer) transgenic mice (SERCA2 KO). Serca2 mRNA transcript abundance was used as a sensitive indicator of Cre-loxP-dependent gene disruption in the myocardium. We found that 2 i.p. injections of tamoxifen in oil (1 mg/day, approximate total dose 80 mg/kg) was sufficient for efficient gene disruption with maximal reduction of Serca2 mRNA as early as 4 days after tamoxifen induction. Moreover, a simple protocol using tamoxifen-supplemented non-pelleted dry feed p.o. was comparable to i.p. injections in inducing gene disruption. These improvements may significantly improve animal welfare and reduce the workload in the production of cardiac conditional knockout mice.

  12. 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.

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

    PubMed Central

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

    2013-01-01

    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 (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 (CBA-flox mice). MHC-HO-1 overexpress the HO-1 gene and enzymatically protein following TAM administration (40 mg/kg body weight on two 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. PMID:23732814

  14. 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.

  15. 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.

  16. Quantitative comparison of cis-regulatory element (CRE) activities in transgenic Drosophila melanogaster.

    PubMed

    Rogers, William A; Williams, Thomas M

    2011-01-01

    Gene expression patterns are specified by cis-regulatory element (CRE) sequences, which are also called enhancers or cis-regulatory modules. A typical CRE possesses an arrangement of binding sites for several transcription factor proteins that confer a regulatory logic specifying when, where, and at what level the regulated gene(s) is expressed. The full set of CREs within an animal genome encodes the organism's program for development, and empirical as well as theoretical studies indicate that mutations in CREs played a prominent role in morphological evolution. Moreover, human genome wide association studies indicate that genetic variation in CREs contribute substantially to phenotypic variation. Thus, understanding regulatory logic and how mutations affect such logic is a central goal of genetics. Reporter transgenes provide a powerful method to study the in vivo function of CREs. Here a known or suspected CRE sequence is coupled to heterologous promoter and coding sequences for a reporter gene encoding an easily observable protein product. When a reporter transgene is inserted into a host organism, the CRE's activity becomes visible in the form of the encoded reporter protein. P-element mediated transgenesis in the fruit fly species Drosophila (D.) melanogaster has been used for decades to introduce reporter transgenes into this model organism, though the genomic placement of transgenes is random. Hence, reporter gene activity is strongly influenced by the local chromatin and gene environment, limiting CRE comparisons to being qualitative. In recent years, the phiC31 based integration system was adapted for use in D. melanogaster to insert transgenes into specific genome landing sites. This capability has made the quantitative measurement of gene and, relevant here, CRE activity feasible. The production of transgenic fruit flies can be outsourced, including phiC31-based integration, eliminating the need to purchase expensive equipment and/or have proficiency at

  17. 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.

  18. 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. PMID:23176440

  19. Evaluation of the Oxoid Brilliance™ CRE Agar for the detection of carbapenemase-producing Enterobacteriaceae.

    PubMed

    Cohen Stuart, J; Voets, G; Rottier, W; Voskuil, S; Scharringa, J; Van Dijk, K; Fluit, A C; Leverstein-Van Hall, M

    2013-11-01

    The adequate detection of carbapenemase-producing Enterobacteriaceae (CPE) is essential for adequate antibiotic therapy and for infection control purposes, especially in an outbreak setting. Selective agars play an important role in the detection of CPE. The Oxoid Brilliance™ CRE Agar (Thermo Fisher Scientific) was evaluated for the detection of CPE using 255 non-repetitive Enterobacteriaceae isolates, including 95 CPE (36 KPC, 4 KPC plus VIM, 4 NDM, 6 GIM, 20 VIM, and 25 OXA-48-producing isolates). The sensitivity of the CRE agar for the detection of CPE was 94 % (89/95), but differed per carbapenemase gene (100 % for KPC, NDM, and GIM, 90 % for VIM, and 84 % for OXA-48-producing isolates). The specificity of the CRE agar was 71 %, due to the growth of AmpC- and/or ESBL-producing isolates. The CRE agar is a sensitive tool for the detection of KPC and metallo-carbapenemase-producing Enterobacteriaceae, although the detection of OXA-48 producers is less optimal. The relatively low specificity requires confirmation of carbapenemase production for isolates recovered from the CRE agar.

  20. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1.

    PubMed Central

    Sauer, B; Henderson, N

    1988-01-01

    The Cre protein encoded by the coliphage P1 is a 38-kDa protein that efficiently promotes both intra- and intermolecular synapsis and recombination of DNA both in Escherichia coli and in vitro. Recombination occurs at a specific site, called lox, and does not require any other protein factors. The Cre protein is shown here also to be able to cause synapsis of DNA and site-specific recombination in a mammalian cell line. A stable mouse cell line was established that expresses the Cre protein under the control of the Cd2+-inducible metallothionein I gene promoter. DNA recombination was monitored with DNA substrates containing two directly repeated lox sites. One such substrate is a circular plasmid with two directly repeated lox sites (lox2) flanking a marker gene and was introduced into cells by Ca3(PO4)2 transformation. As a second substrate we used a pseudorabies virus (a herpesvirus) containing a lox2 insertion designed to provide a sensitive detection system for recombination. In both cases, site-specific recombination in vivo is dependent on the presence of the Cre protein and occurs specifically at the 34-base-pair lox sites. These results demonstrate the controlled site-specific synapsis of DNA and recombination by a prokaryotic protein in mammalian cells and suggest that Cre-mediated site-specific recombination may be a useful tool for understanding and modulating genome rearrangements in eukaryotes. Images PMID:2839833

  1. A BAC-Based Transgenic Mouse Specifically Expresses an Inducible Cre in the Urothelium

    PubMed Central

    Shen, Tian Huai; Gladoun, Nataliya; Castillo-Martin, Mireia; Bonal, Dennis; Domingo-Domenech, Josep; Charytonowicz, Daniel; Cordon-Cardo, Carlos

    2012-01-01

    Cre-loxp mediated conditional knockout strategy has played critical roles for revealing functions of many genes essential for development, as well as the causal relationships between gene mutations and diseases in the postnatal adult mice. One key factor of this strategy is the availability of mice with tissue- or cell type-specific Cre expression. However, the success of the traditional molecular cloning approach to generate mice with tissue specific Cre expression often depends on luck. Here we provide a better alternative by using bacterial artificial chromosome (BAC)-based recombineering to insert iCreERT2 cDNA at the ATG start of the Upk2 gene. The BAC-based transgenic mice express the inducible Cre specifically in the urothelium as demonstrated by mRNA expression and staining for LacZ expression after crossing with a Rosa26 reporter mouse. Taking into consideration the size of the gene of interest and neighboring genes included in a BAC, this method should be widely applicable for generation of mice with tissue specific gene expression or deletions in a more specific manner than previously reported. PMID:22496911

  2. Different thermostabilities of FLP and Cre recombinases: implications for applied site-specific recombination.

    PubMed

    Buchholz, F; Ringrose, L; Angrand, P O; Rossi, F; Stewart, A F

    1996-11-01

    Genomic manipulations using site-specific recombinases rely on their applied characteristics in living systems. To understand their applied properties so that they can be optimally deployed, we compared the recombinases FLP and Cre in two assays. In both Escherichia coli and in vitro, FLP shows a different temperature optimum than Cre. FLP is more thermolabile, having an optimum near 30 degrees C and little detectable activity above 39 degrees C. Cre is optimally efficient at 37 degrees C and above. Consistent with FLP thermolability, recombination in a mammalian cell line mediated by a ligand- regulated FLP-androgen receptor fusion protein is more efficient at 35 degrees C than at higher temperatures. We also document a mutation in a commercially available FLP plasmid (FLP-F70L) which renders this recombinase even more thermolabile. The different temperature optima of FLP, FLP-F70L and Cre influence their strategies of usage. Our results recommend the use of Cre for applications in mice that require efficient recombination. The thermolabilities of FLP and FLP-F70L can be usefully exploited for gain of function and cell culture applications.

  3. 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.

  4. Cell Type-Specific Manipulation with GFP-Dependent Cre Recombinase

    PubMed Central

    Tang, Jonathan C Y; Rudolph, Stephanie; Dhande, Onkar S; Abraira, Victoria E; Choi, Seungwon; Lapan, Sylvain; Drew, Iain R; Drokhlyansky, Eugene; Huberman, Andrew D; Regehr, Wade G; Cepko, Constance L

    2016-01-01

    Summary There are many transgenic GFP reporter lines that allow visualization of specific populations of cells. Using such lines for functional studies requires a method that transforms GFP into a molecule that enables genetic manipulation. Here we report the creation of a method that exploits GFP for gene manipulation, Cre Recombinase Dependent on GFP (CRE-DOG), a split component system that uses GFP and its derivatives to directly induce Cre/loxP recombination. Using plasmid electroporation and AAV viral vectors, we delivered CRE-DOG to multiple GFP mouse lines, leading to effective recombination selectively in GFP-labeled cells. Further, CRE-DOG enabled optogenetic control of these neurons. Beyond providing a new set of tools for manipulation of gene expression selectively in GFP+ cells, we demonstrate that GFP can be used to reconstitute the activity of a protein not known to have a modular structure, suggesting that this strategy might be applicable to a wide range of proteins. PMID:26258682

  5. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands

    PubMed Central

    Maruyama, Eri O.; Aure, Marit H.; Xie, Xiaoling; Myal, Yvonne; Gan, Lin; Ovitt, Catherine E.

    2016-01-01

    The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (Pip) gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts. PMID:26751783

  6. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands.

    PubMed

    Maruyama, Eri O; Aure, Marit H; Xie, Xiaoling; Myal, Yvonne; Gan, Lin; Ovitt, Catherine E

    2016-01-01

    The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (Pip) gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts. PMID:26751783

  7. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands.

    PubMed

    Maruyama, Eri O; Aure, Marit H; Xie, Xiaoling; Myal, Yvonne; Gan, Lin; Ovitt, Catherine E

    2016-01-01

    The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (Pip) gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts.

  8. Neuropilin-2 genomic elements drive cre recombinase expression in primitive blood, vascular and neuronal lineages.

    PubMed

    Wiszniak, Sophie; Scherer, Michaela; Ramshaw, Hayley; Schwarz, Quenten

    2015-11-01

    We have established a novel Cre mouse line, using genomic elements encompassing the Nrp2 locus, present within a bacterial artificial chromosome clone. By crossing this Cre driver line to R26R LacZ reporter mice, we have documented the temporal expression and lineage traced tissues in which Cre is expressed. Nrp2-Cre drives expression in primitive blood cells arising from the yolk sac, venous and lymphatic endothelial cells, peripheral sensory ganglia, and the lung bud. This mouse line will provide a new tool to researchers wishing to study the development of various tissues and organs in which this Cre driver is expressed, as well as allow tissue-specific knockout of genes of interest to study protein function. This work also presents the first evidence for expression of Nrp2 protein in a mesodermal progenitor with restricted hematopoietic potential, which will significantly advance the study of primitive erythropoiesis. genesis 53:709-717, 2015. © 2015 Wiley Periodicals, Inc. PMID:26454009

  9. New Mouse Lines for the Analysis of Neuronal Morphology Using CreER(T)/loxP-Directed Sparse Labeling

    PubMed Central

    Badea, Tudor C.; Hua, Zhong L.; Smallwood, Philip M.; Williams, John; Rotolo, Thomas; Ye, Xin; Nathans, Jeremy

    2009-01-01

    Background Pharmacologic control of Cre-mediated recombination using tamoxifen-dependent activation of a Cre-estrogen receptor ligand binding domain fusion protein [CreER(T)] is widely used to modify and/or visualize cells in the mouse. Methods and Findings We describe here two new mouse lines, constructed by gene targeting to the Rosa26 locus to facilitate Cre-mediated cell modification. These lines should prove particularly useful in the context of sparse labeling experiments. The R26rtTACreER line provides ubiquitous expression of CreER under transcriptional control by the tetracycline reverse transactivator (rtTA); dual control by doxycycline and tamoxifen provides an extended dynamic range of Cre-mediated recombination activity. The R26IAP line provides high efficiency Cre-mediated activation of human placental alkaline phosphatase (hPLAP), complementing the widely used, but low efficiency, Z/AP line. By crossing with mouse lines that direct cell-type specific CreER expression, the R26IAP line has been used to produce atlases of labeled cholinergic and catecholaminergic neurons in the mouse brain. The R26IAP line has also been used to visualize the full morphologies of retinal dopaminergic amacrine cells, among the largest neurons in the mammalian retina. Conclusions The two new mouse lines described here expand the repertoire of genetically engineered mice available for controlled in vivo recombination and cell labeling using the Cre-lox system. PMID:19924248

  10. 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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:26923756

  12. Distracted Driving Raises Crash Risk

    MedlinePlus

    ... Raises Crash Risk Video technology and in-vehicle sensors showed that distracted driving, especially among new drivers, ... whenever the cars were moving. A suite of sensors recorded acceleration, sudden braking or swerving, and other ...

  13. A Resource of Cre Driver Lines for Genetic Targeting of GABAergic Neurons in Cerebral Cortex

    PubMed Central

    Taniguchi, Hiroki; He, Miao; Wu, Priscilla; Kim, Sangyong; Paik, Raehum; Sugino, Ken; Kvitsani, Duda; Fu, Yu; Lu, Jiangteng; Lin, Ying; Miyoshi, Goichi; Shima, Yasuyuki; Fishell, Gord; Nelson, Sacha B.

    2013-01-01

    Summary A key obstacle to understanding neural circuits in the cerebral cortex is that of unraveling the diversity of GABAergic interneurons. This diversity poses general questions for neural circuit analysis: how are these interneuron cell types generated and assembled into stereotyped local circuits and how do they differentially contribute to circuit operations that underlie cortical functions ranging from perception to cognition? Using genetic engineering in mice, we have generated and characterized ~20 Cre and inducible CreER knockin driver lines that reliably target major classes and lineages of GABAergic neurons. More select populations are captured by intersection of Cre and Flp drivers. Genetic targeting allows reliable identification, monitoring, and manipulation of cortical GABAergic neurons, thereby enabling a systematic and comprehensive analysis from cell fate specification, migration, and connectivity, to their functions in network dynamics and behavior. As such, this approach will accelerate the study of GABAergic circuits throughout the mammalian brain. PMID:21943598

  14. Cre-driven optogenetics in the heterogeneous genetic panorama of the VTA.

    PubMed

    Pupe, Stéfano; Wallén-Mackenzie, Åsa

    2015-06-01

    The selectivity of optogenetics commonly relies on genetic promoters to manipulate specific populations of neurons through the use of Cre-driver lines. All studies performed in the ventral tegmental area (VTA) so far have utilized promoters present in groups of cells that release dopamine (DA), GABA, or glutamate. However, neurons that co-release neurotransmitters and variabilities within groups of neurons that release the same neurotransmitter present challenges when evaluating the results. Further complexity is introduced by ectopic expression patterns often occurring in transgenic Cre-drivers. New perspectives could be unfolded by identifying and selecting different types of promoter for driving the Cre recombinase. Here, we discuss some promising candidates and highlight the advantages or disadvantages of different methods for creating novel transgenic lines.

  15. A directional strategy for monitoring Cre-mediated recombination at the cellular level in the mouse.

    PubMed

    Schnütgen, Frank; Doerflinger, Nathalie; Calléja, Cécile; Wendling, Olivia; Chambon, Pierre; Ghyselinck, Norbert B

    2003-05-01

    Functional redundancies, compensatory mechanisms, and lethal phenotypes often prevent the full analysis of gene functions through generation of germline null mutations in the mouse. The use of site-specific recombinases, such as Cre, which catalyzes recombination between loxP sites, has allowed the engineering of mice harboring targeted somatic mutations, which are both temporally controlled and cell-type restricted. Many Cre-expressing mouse lines exist, but only a few transgenic lines are available that harbor a reporter gene whose expression is dependent on a Cre-mediated event. Moreover, their use to monitor gene ablation at the level of individual cells is often limited, as in some tissues the reporter gene may be silenced, be affected by position-effect variegation, or reside in a chromatin configuration inaccessible for recombination. Thus, one cannot validly extrapolate from the expression of a reporter transgene to an identical ablation pattern for the conditional allele of a given gene. By combining the ability of Cre recombinase to invert or excise a DNA fragment, depending on the orientation of the flanking loxP sites, and the availability of both wild-type (WT) and mutant loxP sites, we designed a Cre-dependent genetic switch (FLEx switch) through which the expression of a given gene is turned off, while the expression of another one is concomitantly turned on. We demonstrate the efficiency and reliability of this switch to readily detect, in the mouse, at the single cell level, Cre-mediated gene ablation. We discuss how this strategy can be used to generate genetic modifications in a conditional manner. PMID:12665802

  16. 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. PMID:23050235

  17. 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.

  18. 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. PMID:24747534

  19. Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases.

    PubMed

    Indra, A K; Warot, X; Brocard, J; Bornert, J M; Xiao, J H; Chambon, P; Metzger, D

    1999-11-15

    Conditional DNA excision between two LoxP sites can be achieved in the mouse using Cre-ER(T), a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER ligands. We have recently characterized a new ligand-dependent recombinase, Cre-ER(T2), which was approximately 4-fold more efficiently induced by OHT than Cre-ER(T) in cultured cells. In order to compare the in vivo efficiency of these two ligand-inducible recombinases to generate temporally-controlled somatic mutations, we have engineered transgenic mice expressing a LoxP-flanked (floxed) transgene reporter and either Cre-ER(T) or Cre-ER(T2) under the control of the bovine keratin 5 promoter that is specifically active in the epidermis basal cell layer. No background recombinase activity could be detected, while recombination was induced in basal keratinocytes upon OHT administration. Interestingly, a dose-response study showed that Cre-ER(T2) was approximately 10-fold more sensitive to OHT induction than Cre-ER(T). PMID:10536138

  20. Raising HDL cholesterol in women

    PubMed Central

    Eapen, Danny J; Kalra, Girish L; Rifai, Luay; Eapen, Christina A; Merchant, Nadya; Khan, Bobby V

    2010-01-01

    High-density lipoprotein cholesterol (HDL-C) concentration is essential in the determination of coronary heart disease (CHD) risk in women. This is especially true in the postmenopausal state, where lipid profiles and CHD risk mimic that of age-matched men. Thus, interventions designed to reduce CHD risk by raising HDL-C levels may have particular significance during the transition to menopause. This review discusses HDL-C-raising therapies and the role of HDL in the primary prevention of CHD in women. Lifestyle-based interventions such as dietary change, aerobic exercise regimens, and smoking cessation are initial steps that are effective in raising HDL-C, and available data suggest women respond similarly to men with these interventions. When combined with pharmacotherapy, the effects of these lifestyle alterations are further amplified. Though studies demonstrating gender-specific differences in therapy are limited, niacin continues to be the most effective agent in raising HDL-C levels, especially when used in combination with fibrate or statin therapy. Emerging treatments such as HDL mimetic therapy show much promise in further raising HDL-C levels and improving cardiovascular outcomes. PMID:21072287

  1. Strategic Plan. Volume 2

    NASA Technical Reports Server (NTRS)

    2002-01-01

    The Mission of the NSBRI will be to lead a National effort for accomplishing the integrated, critical path, biomedical research necessary to support the long term human presence, development, and exploration of space and to enhance life on Earth by applying the resultant advances in human knowledge and technology acquired through living and working in space. To carry out this mission, the NSBRI focuses its activities on three Strategic Programs: Strategic Program 1: Countermeasure Research Strategic Program 2: Education, Training and Outreach Strategic Program 3: Cooperative Research and Development. This document contains the detailed Team Strategic Plans for the 11 research teams focused on Strategic Program 1, and the Education and Outreach Team focused on Strategic Program 2. There is overlap and integration among the Programs and Team Strategic Plans, as described in each of the Plans.

  2. Strategic planning and competition

    SciTech Connect

    Gang, W.G. )

    1994-02-01

    This article discusses how to formulate a successful strategic plan in the face of competition from other electric utilities. Areas covered include reasons for plan failure, competitive simulations to test strategic plans, intelligence gathering, and cost reduction through reorganization.

  3. A Hoxa13: Cre Mouse Strain for Conditional Gene Manipulation in Developing Limb, Hindgut, and Urogenital System

    PubMed Central

    Scotti, Martina; Kherdjemil, Yacine; Roux, Marine; Kmita, Marie

    2015-01-01

    SUMMARY The developing limb is a useful model for studying organogenesis and developmental processes. Although Cre alleles exist for conditional loss- or gain-of-function in limbs, Cre alleles targeting specific limb subdomains are desirable. Here we report on the generation of the Hoxa13:Cre line, in which the Cre gene is inserted in the endogenous Hoxa13 gene. We provide evidence that the Cre is active in embryonic tissues/ regions where the endogenous Hoxa13 gene is expressed. Our results show that cells expressing Hoxa13 in developing limb buds contribute to the entire autopod (hand/feet) skeleton and validate Hoxa13 as a distal limb marker as far as the skeleton is concerned. In contrast, in the limb musculature, Cre-based fate mapping shows that almost all muscle masses of the zeugopod (forearm) and part of the triceps contain Hoxa13-expressing cells and/or their descendants. Besides the limb, the activity of the Cre is detectable in the urogenital system and the hindgut, primarily in the epithelium and smooth muscles. Together our data show that the Hoxa13:Cre allele is a useful tool for conditional gene manipulation in the urogenital system, posterior digestive tract, autopod and part of the limb musculature. PMID:25980463

  4. 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.

  5. Mutations in the CRE pocket of bacterial RNA polymerase affect multiple steps of transcription.

    PubMed

    Petushkov, Ivan; Pupov, Danil; Bass, Irina; Kulbachinskiy, Andrey

    2015-07-13

    During transcription, the catalytic core of RNA polymerase (RNAP) must interact with the DNA template with low-sequence specificity to ensure efficient enzyme translocation and RNA extension. Unexpectedly, recent structural studies of bacterial promoter complexes revealed specific interactions between the nontemplate DNA strand at the downstream edge of the transcription bubble (CRE, core recognition element) and a protein pocket formed by core RNAP (CRE pocket). We investigated the roles of these interactions in transcription by analyzing point amino acid substitutions and deletions in Escherichia coli RNAP. The mutations affected multiple steps of transcription, including promoter recognition, RNA elongation and termination. In particular, we showed that interactions of the CRE pocket with a nontemplate guanine immediately downstream of the active center stimulate RNA-hairpin-dependent transcription pausing but not other types of pausing. Thus, conformational changes of the elongation complex induced by nascent RNA can modulate CRE effects on transcription. The results highlight the roles of specific core RNAP-DNA interactions at different steps of RNA synthesis and suggest their importance for transcription regulation in various organisms.

  6. 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

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

    PubMed

    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

    2016-06-20

    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.

  8. 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

  9. Mutations in the CRE pocket of bacterial RNA polymerase affect multiple steps of transcription

    PubMed Central

    Petushkov, Ivan; Pupov, Danil; Bass, Irina; Kulbachinskiy, Andrey

    2015-01-01

    During transcription, the catalytic core of RNA polymerase (RNAP) must interact with the DNA template with low-sequence specificity to ensure efficient enzyme translocation and RNA extension. Unexpectedly, recent structural studies of bacterial promoter complexes revealed specific interactions between the nontemplate DNA strand at the downstream edge of the transcription bubble (CRE, core recognition element) and a protein pocket formed by core RNAP (CRE pocket). We investigated the roles of these interactions in transcription by analyzing point amino acid substitutions and deletions in Escherichia coli RNAP. The mutations affected multiple steps of transcription, including promoter recognition, RNA elongation and termination. In particular, we showed that interactions of the CRE pocket with a nontemplate guanine immediately downstream of the active center stimulate RNA-hairpin-dependent transcription pausing but not other types of pausing. Thus, conformational changes of the elongation complex induced by nascent RNA can modulate CRE effects on transcription. The results highlight the roles of specific core RNAP–DNA interactions at different steps of RNA synthesis and suggest their importance for transcription regulation in various organisms. PMID:25990734

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

    PubMed

    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

  11. Establishment of a Cre recombinase based mutagenesis protocol for markerless gene deletion in Streptococcus suis.

    PubMed

    Koczula, A; Willenborg, J; Bertram, R; Takamatsu, D; Valentin-Weigand, P; Goethe, R

    2014-12-01

    The lack of knowledge about pathogenicity mechanisms of Streptococcus (S.) suis is, at least partially, attributed to limited methods for its genetic manipulation. Here, we established a Cre-lox based recombination system for markerless gene deletions in S. suis serotype 2 with high selective pressure and without undesired side effects.

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

    PubMed

    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.

  13. Diminished CRE-Induced Plasticity is Linked to Memory Deficits in Familial Alzheimer's Disease Mice.

    PubMed

    Bartolotti, Nancy; Segura, Laura; Lazarov, Orly

    2015-01-01

    The mechanism underlying impaired learning and memory in Alzheimer's disease is not fully elucidated. The phosphorylation of cyclic-AMP response element binding protein (pCREB) in the hippocampus is thought to be a critical initiating step in the formation of long-term memories. Here, we tested CRE-driven gene expression following learning in mice harboring the familial Alzheimer's disease-linked APPswe/PS1ΔE9 mutations using CRE-β galactosidase reporter. We show that young adult APPswe/PS1ΔE9 mice exhibit impaired recognition memory and reduced levels of pCREB, and its cofactors CREB binding protein (CBP) and p-300 following a learning task, compared to their wild type littermate counterparts. Impairments in learning-induced activation of CREB in these mice are manifested by reduced CRE-driven gene transcription. Importantly, expression of the CRE-driven immediate early gene, Egr-1 (Zif268) is decreased in the CA1 region of the hippocampus. These studies implicate defective CREB-dependent plasticity in the mechanism underlying learning and memory deficits in Alzheimer's disease. PMID:26682682

  14. The effect of CreA in glucose and xylose catabolism in Aspergillus nidulans.

    PubMed

    Prathumpai, W; McIntyre, M; Nielsen, J

    2004-02-01

    The catabolism of glucose and xylose was studied in a wild type and creA deleted (carbon catabolite de-repressed) strain of Aspergillus nidulans. Both strains were cultivated in bioreactors with either glucose or xylose as the sole carbon source, or in the presence of both sugars. In the cultivations on single carbon sources, it was demonstrated that xylose acted as a carbon catabolite repressor (xylose cultivations), while the enzymes in the xylose utilisation pathway were also subject to repression in the presence of glucose (glucose cultivations). In the wild type strain growing on the sugar mixture, glucose repression of xylose utilisation was observed; with xylose utilisation occurring only after glucose was depleted. This phenomenon was not seen in the creA deleted strain, where glucose and xylose were catabolised simultaneously. Measurement of key metabolites and the activities of key enzymes in the xylose utilisation pathway revealed that xylose metabolism was occurring in the creA deleted strain, even at high glucose concentrations. Conversely, in the wild type strain, activities of the key enzymes for xylose metabolism increased only when the effects of glucose repression had been relieved. Xylose was both a repressor and an inducer of xylanases at the same time. The creA mutation seemed to have pleiotropic effects on carbohydratases and carbon catabolism.

  15. 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.

  16. Tamoxifen-inducible NaV1.8-CreERT2 recombinase activity in nociceptive neurons of dorsal root ganglia.

    PubMed

    Zhao, Jing; Nassar, Mohammed A; Gavazzi, Isabella; Wood, John N

    2006-08-01

    To explore the function of genes expressed in adult mouse nociceptive neurons, we generated heterozygous knock-in mice expressing the tamoxifen-inducible Cre recombinase construct CreERT2 downstream of the Na(V)1.8 promoter. CreERT2 encodes a Cre recombinase (Cre) fused to a mutant estrogen ligand-binding domain (ERT2) that requires the presence of tamoxifen for activity. We have previously shown that heterozygous Na(V)1.8-Cre mice will delete loxP flanked genes specifically in nociceptive sensory neurons from embryonic day 14. We therefore used the same strategy of homologous recombination and mouse generation, substituting the Cre cassette with CreERT2. No functional Cre recombinase activity was found in CreERT2 mice crossed with reporter mice in the absence of tamoxifen. We found that, as with Na(V)1.8-Cre mice, functional Cre recombinase was present in nociceptive sensory neurons after tamoxifen induction in vivo. However, the percentage of dorsal root ganglion (DRG) neurons expressing functional Cre activity was much reduced (<10% of the number found in the Na(V)1.8-Cre mouse). We also examined Cre recombinase activity in sensory neurons in culture. After treatment with 1 muM tamoxifen for 48 h, 15% of DRG neurons showed Cre activity. Na(V)1.8-CreERT2 animals may thus be useful for single cell studies of the functional consequences of gene ablation in culture, but are unlikely to be useful for behavioral studies. PMID:16850455

  17. How (Not) to Raise Money

    ERIC Educational Resources Information Center

    Goeree, Jacob K.; Maasland, Emiel; Onderstal, Sander; Turner, John L.

    2005-01-01

    We show that standard winner-pay auctions are inept fund-raising mechanisms because of the positive externality bidders forgo if they top another's high bid. Revenues are suppressed as a result and remain finite even when bidders value a dollar donated the same as a dollar kept. This problem does not occur in lotteries and all-pay auctions, where…

  18. Fund Raising for University Libraries

    ERIC Educational Resources Information Center

    Eaton, Andrew J.

    1971-01-01

    Library fund raising must be a cooperative effort involving library staff, development office personnel, faculty, president, trustees, and off-campus volunteers, and friends of the university. There are various sources of funds and kinds of approaches to donors which may be tried. Success is not assured, but the potential is great. (6 references)…

  19. 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 Better in Their…

  20. Central American Grandparents Raising Grandchildren

    ERIC Educational Resources Information Center

    Fuller-Thomson, Esme; Minkler, Meredith

    2007-01-01

    This study documents the prevalence and profile of Central American grandparents raising grandchildren in the United States of America. Using nationally representative data from the Census 2000 Supplementary Survey, Central American grandparent caregivers are compared with their noncare-giving peers. Results indicate that nationally an estimated…

  1. Overview of BioCreAtIvE: critical assessment of information extraction for biology

    PubMed Central

    Hirschman, Lynette; Yeh, Alexander; Blaschke, Christian; Valencia, Alfonso

    2005-01-01

    Background The goal of the first BioCreAtIvE challenge (Critical Assessment of Information Extraction in Biology) was to provide a set of common evaluation tasks to assess the state of the art for text mining applied to biological problems. The results were presented in a workshop held in Granada, Spain March 28–31, 2004. The articles collected in this BMC Bioinformatics supplement entitled "A critical assessment of text mining methods in molecular biology" describe the BioCreAtIvE tasks, systems, results and their independent evaluation. Results BioCreAtIvE focused on two tasks. The first dealt with extraction of gene or protein names from text, and their mapping into standardized gene identifiers for three model organism databases (fly, mouse, yeast). The second task addressed issues of functional annotation, requiring systems to identify specific text passages that supported Gene Ontology annotations for specific proteins, given full text articles. Conclusion The first BioCreAtIvE assessment achieved a high level of international participation (27 groups from 10 countries). The assessment provided state-of-the-art performance results for a basic task (gene name finding and normalization), where the best systems achieved a balanced 80% precision / recall or better, which potentially makes them suitable for real applications in biology. The results for the advanced task (functional annotation from free text) were significantly lower, demonstrating the current limitations of text-mining approaches where knowledge extrapolation and interpretation are required. In addition, an important contribution of BioCreAtIvE has been the creation and release of training and test data sets for both tasks. There are 22 articles in this special issue, including six that provide analyses of results or data quality for the data sets, including a novel inter-annotator consistency assessment for the test set used in task 2. PMID:15960821

  2. Mouse alpha1(I)-collagen promoter is the best known promoter to drive efficient Cre recombinase expression in osteoblast.

    PubMed

    Dacquin, Romain; Starbuck, Michael; Schinke, Thorsten; Karsenty, Gérard

    2002-06-01

    Cell- and time-specific gene inactivation should enhance our knowledge of bone biology. Implementation of this technique requires construction of transgenic mouse lines expressing Cre recombinase in osteoblasts, the bone forming cell. We tested several promoter fragments for their ability to drive efficient Cre expression in osteoblasts. In the first mouse transgenic line, the Cre gene was placed under the control of the 2.3-kb proximal fragment of the alpha1(I)-collagen promoter, which is expressed at high levels in osteoblasts throughout their differentiation. Transgenic mice expressing this transgene in bone were bred with the ROSA26 reporter (R26R) strain in which the ROSA26 locus is targeted with a conditional LacZ reporter cassette. In R26R mice, Cre expression and subsequent Cre-mediated recombination lead to expression of the LacZ reporter gene, an event that can be monitored by LacZ staining. LacZ staining was detected in virtually all osteoblasts of alpha1(I)-Cre;R26R mice indicating that homologous recombination occurred in these cells. No other cell type stained blue. In the second line studied, the 1.3-kb fragment of osteocalcin gene 2 (OG2) promoter, which is active in differentiated osteoblasts, was used to drive Cre expression. OG2-Cre mice expressed Cre specifically in bone. However, cross of OG2-Cre mice with R26R mice did not lead to any detectable LacZ staining in osteoblasts. Lastly, we tested a more active artificial promoter derived from the OG2 promoter. The artificial OG2-Cre transgene was expressed by reverse transcriptase-polymerase chain reaction in cartilage and bone samples. After cross of the artificial OG2-Cre mice with R26R mice, we detected a LacZ staining in articular chondrocytes but not in osteoblasts. Our data suggest that the only promoter able to drive Cre expression at a level sufficient to induce recombination in osteoblasts is the alpha1(I)-collagen promoter. PMID:12112477

  3. 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.

  4. The generation and characterization of novel Col1a1FRT-Cre-ER-T2-FRT and Col1a1FRT-STOP-FRT-Cre-ER-T2 mice for sequential mutagenesis.

    PubMed

    Zhang, Minsi; Kirsch, David G

    2015-09-01

    Novel genetically engineered mouse models using the Cre-loxP or the Flp-FRT systems have generated useful reagents to manipulate the mouse genome in a temporally-regulated and tissue-specific manner. By incorporating a constitutive Cre driver line into a mouse model in which FRT-regulated genes in other cell types are regulated by Flp-FRT recombinase, gene expression can be manipulated simultaneously in separate tissue compartments. This application of dual recombinase technology can be used to dissect the role of stromal cells in tumor development and cancer therapy. Generating mice in which Cre-ER(T2) is expressed under Flp-FRT-mediated regulation would enable step-wise manipulation of the mouse genome using dual recombinase technology. Such next-generation mouse models would enable sequential mutagenesis to better model cancer and define genes required for tumor maintenance. Here, we generated novel genetically engineered mice that activate or delete Cre-ER(T2) in response to Flp recombinase. To potentially utilize the large number of Cre-loxP-regulated transgenic alleles that have already been targeted into the Rosa26 locus, such as different reporters and mutant genes, we targeted the two novel Cre-ER(T2) alleles into the endogenous Col1a1 locus for ubiquitous expression. In the Col1a1(FRT-Cre-ER-T2-FRT) mice, Flp deletes Cre-ER(T2), so that Cre-ER(T2) is only expressed in cells that have never expressed Flp. In contrast, in the Col1a1(FRT-STOP-FRT-Cre-ER-T2) mice, Flp removes the STOP cassette to allow Cre-ER(T2) expression so that Cre-ER(T2) is only expressed in cells that previously expressed Flp. These two new novel mouse strains will be complementary to each other and will enable the exploration of complex biological questions in development, normal tissue homeostasis and cancer. PMID:26183214

  5. Generation of a Transgenic Mouse Model With Chondrocyte-Specific and Tamoxifen-Inducible Expression of Cre Recombinase

    PubMed Central

    Chen, Mo; Lichtler, Alexander C.; Sheu, Tzong-Jen; Xie, Chao; Zhang, Xinping; O’Keefe, Regis J.; Chen, Di

    2009-01-01

    Summary Postnatal cartilage development and growth are regulated by key growth factors and signaling molecules. To fully understand the function of these regulators, an inducible and chondrocyte-specific gene deletion system needs to be established to circumvent the perinatal lethality. In this report, we have generated a transgenic mouse model (Col2a1-CreERT2) in which expression of the Cre recombinase is driven by the chondrocyte-specific col2a1 promoter in a tamoxifen-inducible manner. To determine the specificity and efficiency of the Cre recombination, we have bred Col2a1-CreERT2 mice with Rosa26R reporter mice. The X-Gal staining showed that the Cre recombination is specifically achieved in cartilage tissues with tamoxifen-induction. In vitro experiments of chondrocyte cell culture also demonstrate the 4-hydroxy tamoxifen-induced Cre recombination. These results demonstrate that Col2a1-CreERT2 transgenic mice can be used as a valuable tool for an inducible and chondrocyte-specific gene deletion approach. PMID:17211877

  6. 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.

  7. The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation

    PubMed Central

    2011-01-01

    Background The identification and characterization of the transcriptional regulatory networks governing the physiology and adaptation of microbial cells is a key step in understanding their behaviour. One such wide-domain regulatory circuit, essential to all cells, is carbon catabolite repression (CCR): it allows the cell to prefer some carbon sources, whose assimilation is of high nutritional value, over less profitable ones. In lower multicellular fungi, the C2H2 zinc finger CreA/CRE1 protein has been shown to act as the transcriptional repressor in this process. However, the complete list of its gene targets is not known. Results Here, we deciphered the CRE1 regulatory range in the model cellulose and hemicellulose-degrading fungus Trichoderma reesei (anamorph of Hypocrea jecorina) by profiling transcription in a wild-type and a delta-cre1 mutant strain on glucose at constant growth rates known to repress and de-repress CCR-affected genes. Analysis of genome-wide microarrays reveals 2.8% of transcripts whose expression was regulated in at least one of the four experimental conditions: 47.3% of which were repressed by CRE1, whereas 29.0% were actually induced by CRE1, and 17.2% only affected by the growth rate but CRE1 independent. Among CRE1 repressed transcripts, genes encoding unknown proteins and transport proteins were overrepresented. In addition, we found CRE1-repression of nitrogenous substances uptake, components of chromatin remodeling and the transcriptional mediator complex, as well as developmental processes. Conclusions Our study provides the first global insight into the molecular physiological response of a multicellular fungus to carbon catabolite regulation and identifies several not yet known targets in a growth-controlled environment. PMID:21619626

  8. 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

  9. The Dmp1-SOST Transgene Interacts With and Downregulates the Dmp1-Cre Transgene and the Rosa(Notch) Allele.

    PubMed

    Zanotti, Stefano; Canalis, Ernesto

    2016-05-01

    Activation of Notch1 in osteocytes of Rosa(Notch) mice, where a loxP-flanked STOP cassette and the Nicd coding sequence were targeted to the reverse orientation splice acceptor (Rosa)26 locus, causes osteopetrosis associated with suppressed Sost expression and enhanced Wnt signaling. To determine whether Sost downregulation mediates the effects of Notch activation in osteocytes, Rosa(Notch) mice were crossed with transgenics expressing Cre recombinase or SOST under the control of the dentin matrix protein (Dmp)1 promoter. Dmp1-SOST transgenics displayed vertebral osteopenia and a modest femoral cancellous and cortical bone phenotype, whereas hemizygous Dmp1-Cre transgenics heterozygous for the Rosa(Notch) allele (Dmp1-Cre;Rosa(Notch)) exhibited osteopetrosis. The phenotype of Notch activation in osteocytes was prevented in Dmp1-Cre;Rosa(Notch) mice hemizygous for the Dmp1-SOST transgene. The effect was associated with downregulated Notch signaling and suppressed Dmp1 and Rosa26 expression. To test whether SOST regulates Notch expression in osteocytes, cortical bone cultures from Dmp1-Cre;Rosa(Notch) mice or from Rosa(Notch) control littermates were exposed to recombinant human SOST. The addition of SOST had only modest effects on Notch target gene mRNA levels and suppressed Dmp1, but not Cre or Rosa26, expression. These findings suggest that prevention of the Dmp1-Cre;Rosa(Notch) skeletal phenotype by Dmp1-SOST is not secondary to SOST expression but to interactions among the Dmp1-SOST and Dmp1-Cre transgenes and the Rosa26 locus. In conclusion, the Dmp1-SOST transgene suppresses the expression of the Dmp1-Cre transgene and of Rosa26. PMID:26456319

  10. 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. PMID:27226768

  11. Yeast osmosensor Sln1 and plant cytokinin receptor Cre1 respond to changes in turgor pressure.

    PubMed

    Reiser, VladimIr; Raitt, Desmond C; Saito, Haruo

    2003-06-23

    Very little is known about how cellular osmosensors monitor changes in osmolarity of the environment. Here, we report that in yeast, Sln1 osmosensor histidine kinase monitors changes in turgor pressures. Reductions in turgor caused by either hyperosmotic stress, nystatin, or removal of cell wall activate MAPK Hog1 specifically through the SLN1 branch, but not through the SHO1 branch of the high osmolarity glycerol pathway. The integrity of the periplasmic region of Sln1 was essential for its sensor function. We found that activity of the plant histidine kinase cytokinin response 1 (Cre1) is also regulated by changes in turgor pressure, in a manner identical to that of Sln1, in the presence of cytokinin. We propose that Sln1 and Cre1 are turgor sensors, and that similar turgor-sensing mechanisms might regulate hyperosmotic stress responses both in yeast and plants.

  12. 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.

  13. Utilizing the CreA Gene to Better Understand Carbon Flux in Fungi

    SciTech Connect

    Annecharico, Matthew D.; Baker, Scott E.

    2005-01-01

    Carbon catabolite repression (CCR) is a system which represses the synthesis of enzymes required for certain carbon sources when more favored carbon sources are present. Fungal enzymes are useful in various industrial applications such as the degradation of bi-product biomass produced in many areas of manufacturing and production. For example, the basidiomycete, Phanerochaete chrysosporium, a filamentous fungus, produces a number of lignin peroxidases (LiPs) such as an extra-cellular lignin-modifying protein and manganese-dependent peroxidase (MnPs). The LiPs secreted by P. chrysosporium are able to break down lignin, one of the main substances of wood, while leaving the cellulose behind. These enzymes are capable of degrading toxic waste as well as biomass waste. Through polysaccharide hydrolysis and lignin utilization, biobased products and biofuels can be processed from biomass instead of petroleum, which is more environmentally sound than current fuel options. Previous research in the fungal phylum, Ascomyces has shown that the creA gene is an important gene that controls various enzymes which have key roles in controlling carbon utilization by fungi. We hypothesize that there are similarities across all fungi in the mechanism by which carbon flux is controlled and that orthologs of the creA gene exist in all fungal phyla. A COnsensus-DEgenerate Hybrid Oligonucleotide Primers (CODEHOP) polymerase chain reaction (PCR) strategy was used to isolate genomic fragments of the creA gene from a variety of fungi with the objective to design and utilize a method for isolation of the creA gene from a wide range of fungi.

  14. 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

  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. 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.

  17. 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.

  18. 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. PMID:22902108

  19. 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.

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

    PubMed

    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

  1. Engineering mouse chromosomes with Cre-loxP: range, efficiency, and somatic applications.

    PubMed

    Zheng, B; Sage, M; Sheppeard, E A; Jurecic, V; Bradley, A

    2000-01-01

    Chromosomal rearrangements are important resources for genetic studies. Recently, a Cre-loxP-based method to introduce defined chromosomal rearrangements (deletions, duplications, and inversions) into the mouse genome (chromosome engineering) has been established. To explore the limits of this technology systematically, we have evaluated this strategy on mouse chromosome 11. Although the efficiency of Cre-loxP-mediated recombination decreases with increasing genetic distance when the two endpoints are on the same chromosome, the efficiency is not limiting even when the genetic distance is maximized. Rearrangements encompassing up to three quarters of chromosome 11 have been constructed in mouse embryonic stem (ES) cells. While larger deletions may lead to ES cell lethality, smaller deletions can be produced very efficiently both in ES cells and in vivo in a tissue- or cell-type-specific manner. We conclude that any chromosomal rearrangement can be made in ES cells with the Cre-loxP strategy provided that it does not affect cell viability. In vivo chromosome engineering can be potentially used to achieve somatic losses of heterozygosity in creating mouse models of human cancers. PMID:10611243

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

    PubMed

    Prados, Alejandro; Kollias, George; Koliaraki, Vasiliki

    2016-09-08

    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.

  3. Fabp4-CreER lineage tracing revealstwo distinctive coronary vascular populations

    PubMed Central

    He, Lingjuan; Tian, Xueying; Zhang, Hui; Wythe, Joshua D; Zhou, Bin

    2014-01-01

    Over the last two decades, genetic lineage tracing has allowed for the elucidation of the cellular origins and fates during both embryogenesis and in pathological settings in adults. Recent lineage tracing studies using Apln-CreER tool indicated that a large number of post-natal coronary vessels do not form from pre-existing vessels. Instead, they form de novo after birth, which represents a coronary vascular population (CVP) distinct from the pre-existing one. Herein, we present new coronary vasculature lineage tracing results using a novel tool, Fabp4-CreER. Our results confirm the distinct existence of two unique CVPs. The 1st CVP, which is labelled by Fabp4-CreER, arises through angiogenic sprouting of pre-existing vessels established during early embryogenesis. The 2nd CVP is not labelled by Fabp4, suggesting that these vessels form de novo, rather than through expansion of the 1st CVP. These results support the de novo formation of vessels in the post-natal heart, which has implications for studies in cardiovascular disease and heart regeneration. PMID:25265869

  4. 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

  5. Generation of a cre recombinase-conditional Nos1ap over-expression transgenic mouse

    PubMed Central

    Auer, Dallas R.; Sysa-Shah, Polina; Bedja, Djahida; Simmers, Jessica L.; Pak, Evgenia; Dutra, Amalia; Cohn, Ronald; Gabrielson, Kathleen L.

    2016-01-01

    Polymorphic non-coding variants at the NOS1AP locus have been associated with the common cardiac, metabolic and neurological traits and diseases. Although, in vitro gene targeting-based cellular and biochemical studies have shed some light on NOS1AP function in cardiac and neuronal tissue, to enhance our understanding of NOS1AP function in mammalian physiology and disease, we report the generation of cre recombinase-conditional Nos1ap over-expression transgenic mice (Nos1apTg). Conditional transgenic mice were generated by the pronuclear injection method and three independent, single-site, multiple copies integration event-based founder lines were selected. For heart-restricted over-expression, Nos1apTg mice were crossed with Mlc2v-cre and Nos1ap transcript over-expression was observed in left ventricles from Nos1apTg; Mlc2v-cre F1 mice. We believe that with the potential of conditional over-expression, Nos1apTg mice will be a useful resource in studying NOS1AP function in various tissues under physiological and disease states. PMID:24563304

  6. 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

  7. Loss of Vascular Endothelial Growth Factor A (VEGFA) Isoforms in Granulosa Cells Using pDmrt-1-Cre or Amhr2-Cre Reduces Fertility by Arresting Follicular Development and by Reducing Litter Size in Female Mice

    PubMed Central

    Sargent, Kevin M.; Lu, Ningxia; Clopton, Debra T.; Pohlmeier, William E.; Brauer, Vanessa M.; Ferrara, Napoleone; Silversides, David W.; Cupp, Andrea S.

    2015-01-01

    Because VEGFA has been implicated in follicle development, the objective of this study was to determine the effects of granulosa- and germ cell-specific VEGFA loss on ovarian morphogenesis, function, and female fertility. pDmrt1-Cre mice were mated to floxed VEGFA mice to develop granulosa-/germ cell-specific knockouts (pDmrt1-Cre;Vegfa-/-). The time from mating to first parturition was increased when pDmrt1-Cre;Vegfa-/- females were mated to control males (P = 0.0008) and tended to be longer for heterozygous females (P < 0.07). Litter size was reduced for pDmrt1-Cre;Vegfa-/- females (P < 0.007). The time between the first and second parturitions was also increased for heterozygous females (P < 0.04) and tended to be increased for pDmrt1-Cre;Vegfa-/- females (P < 0.07). pDmrt1-Cre;Vegfa-/- females had smaller ovaries (P < 0.04), reduced plasma estradiol (P < 0.007), fewer developing follicles (P < 0.008) and tended to have fewer corpora lutea (P < 0.08). Expression of Igf1r was reduced (P < 0.05); expression of Foxo3a tended to be increased (P < 0.06); and both Fshr (P < 0.1) and Sirt6 tended to be reduced (P < 0.06) in pDmrt1-Cre;Vegfa-/- ovaries. To compare VEGFA knockouts, we generated Amhr2-Cre;Vegfa-/- mice that required more time from mating to first parturition (P < 0.003) with variable ovarian size. Both lines had more apoptotic granulosa cells, and vascular staining did not appear different. Taken together these data indicate that the loss of all VEGFA isoforms in granulosa/germ cells (proangiogenic and antiangiogenic) causes subfertility by arresting follicular development, resulting in reduced ovulation rate and fewer pups per litter. PMID:25658474

  8. Generation and Characterisation of a Pax8-CreERT2 Transgenic Line and a Slc22a6-CreERT2 Knock-In Line for Inducible and Specific Genetic Manipulation of Renal Tubular Epithelial Cells.

    PubMed

    Espana-Agusti, Judit; Zou, Xiangang; Wong, Kim; Fu, Beiyuan; Yang, Fengtang; Tuveson, David A; Adams, David J; Matakidou, Athena

    2016-01-01

    Genetically relevant mouse models need to recapitulate the hallmarks of human disease by permitting spatiotemporal gene targeting. This is especially important for replicating the biology of complex diseases like cancer, where genetic events occur in a sporadic fashion within developed somatic tissues. Though a number of renal tubule targeting mouse lines have been developed their utility for the study of renal disease is limited by lack of inducibility and specificity. In this study we describe the generation and characterisation of two novel mouse lines directing CreERT2 expression to renal tubular epithelia. The Pax8-CreERT2 transgenic line uses the mouse Pax8 promoter to direct expression of CreERT2 to all renal tubular compartments (proximal and distal tubules as well as collecting ducts) whilst the Slc22a6-CreERT2 knock-in line utilises the endogenous mouse Slc22a6 locus to specifically target the epithelium of proximal renal tubules. Both lines show high organ and tissue specificity with no extrarenal activity detected. To establish the utility of these lines for the study of renal cancer biology, Pax8-CreERT2 and Slc22a6-CreERT2 mice were crossed to conditional Vhl knockout mice to induce long-term renal tubule specific Vhl deletion. These models exhibited renal specific activation of the hypoxia inducible factor pathway (a VHL target). Our results establish Pax8-CreERT2 and Slc22a6-CreERT2 mice as valuable tools for the investigation and modelling of complex renal biology and disease. PMID:26866916

  9. A transgenic Tbx6;CreERT2 line for inducible gene manipulation in the presomitic mesoderm.

    PubMed

    Peter Lopez, T; Fan, Chen-Ming

    2012-06-01

    The rhythmic segmentation process of the presomitic mesoderm (PSM) orchestrates the formation of somites, the fundamental units for the vertebrate axial body plan. To aid the investigation of molecular components governing the conversion from PSM into somites, we generated a transgenic mouse line that expresses a tamoxifen (tmx) inducible CreER(T2) under the control of a 2.5 kb enhancer element of Tbx6, a gene essential for PSM formation and somite patterning. Combined with Cre reporters, this Tbx6;CreER(T2) line displays robust tmx-inducible Cre activity in the PSM at various embryonic stages. This tool should be useful for studying gene function during somitogenesis by either conditional inactivation or mis-expression, and potentially coupled with cell marking.

  10. Quality control of astrocyte-directed Cre transgenic mice: the benefits of a direct link between loss of gene expression and reporter activation.

    PubMed

    Requardt, Robert Pascal; Kaczmarczyk, Lech; Dublin, Pavel; Wallraff-Beck, Anke; Mikeska, Thomas; Degen, Joachim; Waha, Andreas; Steinhäuser, Christian; Willecke, Klaus; Theis, Martin

    2009-04-15

    Cre recombinase activity for cell-type restricted deletion of floxed target genes (i.e., flanked by Cre recognition loxP-sites) is often measured by separate matings with recombination-activated reporter gene mice. Using a floxed Gja1 (Cx43) allele, we demonstrate the benefits of a direct link between reporter gene expression and target gene deletion to overcome critical limitations of the Cre/loxP system. The widely used human glial fibrillary acidic protein (hGFAP)-Cre transgene exhibits variable recombination activity and requires postexperimental validation. Such quality control is essential to correlate the extent of Cre-mediated Gja1 ablation with phenotypical alterations and to maintain the activity status of hGFAP-Cre in transgenic mouse colonies. We present several strategies to control for the fidelity of hGFAP-Cre mediated recombination. (c) 2008 Wiley-Liss, Inc.

  11. Honeybee royal jelly and nobiletin stimulate CRE-mediated transcription in ERK-independent and -dependent fashions, respectively, in PC12D cells.

    PubMed

    Fujiwara, Hironori; Kogure, Ako; Sakamoto, Masahiro; Yamakuni, Tohru; Mimaki, Yoshihiro; Murata, Kiyoshi; Hitomi, Nobuyuki; Yamaguchi, Kikuji; Ohizumi, Yasushi

    2011-01-01

    To prove the pharmacological actions of honeybee royal jelly (RJ) on the nervous system, we examined the effects of RJ on CRE-mediated transcription. RJ increased CRE-mediated transcription in PC12D cells. Moreover, CRE-mediated transcriptional activity by RJ was enhanced by nobiletin. U0126, a MEK inhibitor, inhibited CRE-mediated transcription by combining RJ plus nobiletin without affecting transcription by RJ alone. These results suggest that RJ stimulates CRE-mediated transcription via an ERK-independent cascade, whereas the increasing CRE-mediated transcriptional effect by nobiletin is dependent on ERK phosphorylation. Combining RJ plus nobiletin may activate effectively neuronal functions via enhancement of CRE-mediated transcription.

  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. Simple and efficient recycling of fungal selectable marker genes with the Cre-loxP recombination system via anastomosis

    PubMed Central

    Zhang, Dong-Xiu; Lu, Hsiao-Ling; Liao, Xinggang; St. Leger, Raymond J.; Nuss, Donald L.

    2013-01-01

    Reverse-genetics analysis has played a significant role in advancing fungal biology, but is limited by the number of available selectable marker genes (SMGs). The Cre-loxP recombination system has been adapted for use in filamentous fungi to overcome this limitation. Expression of the Cre recombinase results in excision of an integrated SMG that is flanked by loxP sites, allowing a subsequent round of transformation with the same SMG. However, current protocols for regulated expression or presentation of Cre require multiple time-consuming steps. During efforts to disrupt four different RNA-dependent RNA polymerase genes in a single strain of the chestnut blight fungus Cryphonectria parasitica, we tested whether Cre could successfully excise loxP-flanked SMGs when provided in trans via anastomosis. Stable Cre-producing donor strains were constructed by transformation of wild-type C. parasitica strain EP155 with the Cre-coding domain under the control of a constitutive promoter. Excision of multiple loxP-flanked SMGs was efficiently achieved by simply pairing the Cre-donor strain and the loxP-flanked SMGs-transformed recipient strain and recovering mycelia from the margin of the recipient colony near the anastomosis zone. This method was shown to be as efficient as and much less time consuming than excision by transformation-mediated expression of Cre. It also allows unlimited recycling of loxP-flanked SMGs and the generation of disruption mutant strains that are free of any foreign gene. The successful application of this method to Metarhizium robertsii suggests potential use for optimizing reverse-genetics analysis in a broad range of filamentous fungi. PMID:24007936

  14. 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.

  15. Strategic Initiatives and Goals.

    ERIC Educational Resources Information Center

    Howard Community Coll., Columbia, MD.

    This document outlines Howard Community College's (HCC) six strategic initiatives and goals. Each of the strategic initiatives is presented, along with a context for the statement and a list of goals to be achieved in support of the overall initiative. First, HCC will be a learning community that provides possibilities for learning that address…

  16. 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.

  17. 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…

  18. 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…

  19. 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.

  20. A Tlx2-Cre mouse line uncovers essential roles for hand1 in extraembryonic and lateral mesoderm.

    PubMed

    Maska, Emily L; Cserjesi, Peter; Hua, Lisa L; Garstka, Meghan E; Brody, Heather M; Morikawa, Yuka

    2010-08-01

    Hand1 regulates development of numerous tissues within the embryo, extraembryonic mesoderm, and trophectoderm. Systemic loss of Hand1 results in early embryonic lethality but the cause has remained unknown. To determine if Hand1 expression in extraembryonic mesoderm is essential for embryonic survival, Hand1 was conditionally deleted using the HoxB6-Cre mouse line that expresses Cre in extraembryonic and lateral mesoderm. Deletion of Hand1 using HoxB6-Cre resulted in embryonic lethality identical to systemic knockout. To determine if lethality is due to Hand1 function in extraembryonic mesoderm or lateral mesoderm, we generated a Tlx2-Cre mouse line expressing Cre in lateral mesoderm but not extraembryonic tissues. Deletion of Hand1 using the Tlx2-Cre line results in embryonic survival with embryos exhibiting herniated gut and thin enteric smooth muscle. Our results show that Hand1 regulates development of lateral mesoderm derivatives and its loss in extraembryonic mesoderm is the primary cause of lethality in Hand1-null embryos.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Cre-mediated stress affects sirtuin expression levels, peroxisome biogenesis and metabolism, antioxidant and proinflammatory signaling pathways.

    PubMed

    Xiao, Yu; Karnati, Srikanth; Qian, Guofeng; Nenicu, Anca; Fan, Wei; Tchatalbachev, Svetlin; Höland, Anita; Hossain, Hamid; Guillou, Florian; Lüers, Georg H; Baumgart-Vogt, Eveline

    2012-01-01

    Cre-mediated excision of loxP sites is widely used in mice to manipulate gene function in a tissue-specific manner. To analyze phenotypic alterations related to Cre-expression, we have used AMH-Cre-transgenic mice as a model system. Different Cre expression levels were obtained by investigation of C57BL/6J wild type as well as heterozygous and homozygous AMH-Cre-mice. Our results indicate that Cre-expression itself in Sertoli cells already has led to oxidative stress and lipid peroxidation (4-HNE lysine adducts), inducing PPARα/γ, peroxisome proliferation and alterations of peroxisome biogenesis (PEX5, PEX13 and PEX14) as well as metabolic proteins (ABCD1, ABCD3, MFP1, thiolase B, catalase). In addition to the strong catalase increase, a NRF2- and FOXO3-mediated antioxidative response (HMOX1 of the endoplasmic reticulum and mitochondrial SOD2) and a NF-κB activation were noted. TGFβ1 and proinflammatory cytokines like IL1, IL6 and TNFα were upregulated and stress-related signaling pathways were induced. Sertoli cell mRNA-microarray analysis revealed an increase of TNFR2-signaling components. 53BP1 recruitment and expression levels for DNA repair genes as well as for p53 were elevated and the ones for related sirtuin deacetylases affected (SIRT 1, 3-7) in Sertoli cells. Under chronic Cre-mediated DNA damage conditions a strong downregulation of Sirt1 was observed, suggesting that the decrease of this important coordinator between DNA repair and metabolic signaling might induce the repression release of major transcription factors regulating metabolic and cytokine-mediated stress pathways. Indeed, caspase-3 was activated and increased germ cell apoptosis was observed, suggesting paracrine effects. In conclusion, the observed wide stress-induced effects and metabolic alterations suggest that it is essential to use the correct control animals (Cre/Wt) with matched Cre expression levels to differentiate between Cre-mediated and specific gene-knock out

  6. 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.

  7. 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.”

  8. Successful Processes to Engender Board Ownership of Strategic Planning

    ERIC Educational Resources Information Center

    Calareso, Jack P.

    2007-01-01

    Too often, Boards of Trustees feel that final approval of the institution's Strategic Plan is sufficient. However, most Plans will only be successfully implemented if the Board has complete ownership and a commitment to be actively involved in many areas of the institution (not only fund raising). This article describes a successful process and…

  9. 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…

  10. lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets.

    PubMed

    Carow, Berit; Gao, Yu; Coquet, Jonathan; Reilly, Marie; Rottenberg, Martin E

    2016-09-15

    Conditional gene targeting using the bacteriophage-derived Cre recombinase is widely applied for functional gene studies in mice. Mice transgenic for Cre under the control of the lck gene promoter are used to study the role of loxP-targeted genes in T cell development and function. In this article, we show a striking 65% reduction in cellularity, preferential development of γδ versus αβ T cells, and increased expression of IL-7R in the thymus of mice expressing Cre under the proximal lck promoter (lck-cre(+) mice). The transition from CD4/CD8 double-negative to double-positive cells was blocked, and lck-cre(+) double-positive cells were more prone to apoptosis and showed higher levels of Cre expression. Importantly, numbers of naive T cells were reduced in spleens and lymph nodes of lck-cre(+) mice. In contrast, frequencies of γδ T cells, CD44(+)CD62L(-) effector T cells, and Foxp3(+) regulatory T cells were elevated, as was the frequency of IFN-γ-secreting CD4(+) and CD8(+) T cells. A literature survey of 332 articles that used lck-cre(+) mice for deletion of floxed genes indicated that results are statistically influenced by the control used (lck-cre(+) or lck-cre(-)), more frequently resembling the lck-cre(+) phenotype described in this article if lck-cre(-) controls were used. Altogether, care should be taken when interpreting published results and to properly control targeted gene deletions using the lck-cre(+) strain. PMID:27503210

  11. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Raised. 60.123 Section 60.123 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards, Inspections... AND SHELLFISH General Provisions Definitions § 60.123 Raised. Raised means in the case of: (a)...

  12. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Raised. 60.123 Section 60.123 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards, Inspections... AND SHELLFISH General Provisions Definitions § 60.123 Raised. Raised means in the case of: (a)...

  13. 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,…

  14. 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,…

  15. Management of raised intracranial pressure.

    PubMed

    Pickard, J D; Czosnyka, M

    1993-08-01

    based on experience of the particular cause of raised intracranial pressure in a given age group. PMID:8350099

  16. Benchmarking for strategic action.

    PubMed

    Jennings, K; Westfall, F

    1992-01-01

    By focusing on three key elements--customer expectations, competitor strengths and vulnerabilities, and organizational competencies--a company's benchmarking effort can be designed to drive the strategic planning process.

  17. Strategic agility for nursing leadership.

    PubMed

    Shirey, Maria R

    2015-06-01

    This department highlights change management strategies that may be successful in strategically planning and executing organizational change. In this article, the author discusses strategic agility as an important leadership competency and offers approaches for incorporating strategic agility in healthcare systems. A strategic agility checklist and infrastructure-building approach are presented. PMID:26010278

  18. The Concept of Strategic Decisionmaking.

    ERIC Educational Resources Information Center

    Collier, Douglas J.

    Strategic decision-making literature is reviewed, and applications to colleges and universities are made. The key requirement for strategic decision-making is that decisions affect the entire organization. While strategic decision-making can occur at different levels within the organization, the specific strategic decisions available to the…

  19. Strategic agility for nursing leadership.

    PubMed

    Shirey, Maria R

    2015-06-01

    This department highlights change management strategies that may be successful in strategically planning and executing organizational change. In this article, the author discusses strategic agility as an important leadership competency and offers approaches for incorporating strategic agility in healthcare systems. A strategic agility checklist and infrastructure-building approach are presented.

  20. 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.

  1. Generation and Characterization of an Nse-CreERT2 Transgenic Line Suitable for Inducible Gene Manipulation in Cerebellar Granule Cells

    PubMed Central

    Pohlkamp, Theresa; Steller, Laura; May, Petra; Günther, Thomas; Schüle, Roland; Frotscher, Michael

    2014-01-01

    We created an Nse-CreERT2 mouse line expressing the tamoxifen-inducible CreERT2 recombinase under the control of the neuron-specific enolase (Nse) promoter. By using Cre reporter lines we could show that this Nse-CreERT2 line has recombination activity in the granule cells of all cerebellar lobules as well as in postmitotic granule cell precursors in the external granular layer of the developing cerebellum. A few hippocampal dentate gyrus granule cells showed Cre-mediated recombination as well. Cre activity could be induced in both the developing and adult mouse brain. The established mouse line constitutes a valuable tool to study the function of genes expressed by cerebellar granule cells in the developing and adult brain. In combination with reporter lines it is a useful model to analyze the development and maintenance of the cerebellar architecture including granule cell distribution, migration, and the extension of granule cell fibers in vivo. PMID:24950299

  2. Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors.

    PubMed

    Vega, Juan M; Yu, Weichang; Han, Fangpu; Kato, Akio; Peters, Eric M; Zhang, Zhanyuan J; Birchler, James A

    2008-04-01

    The Cre/loxP site-specific recombination system has been applied in various plant species including maize (Zea mays) for marker gene removal, gene targeting, and functional genomics. A BIBAC vector system was adapted for maize transformation with a large fragment of genetic material including a herbicide resistance marker gene, a 30 kb yeast genomic fragment as a marker for fluorescence in situ hybridization (FISH), and a 35S-lox-cre recombination cassette. Seventy-five transgenic lines were generated from Agrobacterium-mediated transformation of a maize Hi II line with multiple B chromosomes. Eighty-four inserts have been localized among all 10 A chromosome pairs by FISH using the yeast DNA probe together with a karyotyping cocktail. No inserts were found on the B chromosomes; thus a bias against the B chromosomes by the Agrobacterium-mediated transformation was revealed. The expression of a cre gene was confirmed in 68 of the 75 transgenic lines by a reporter construct for cre/lox mediated recombination. The placement of the cre/lox site-specific recombination system in many locations in the maize genome will be valuable materials for gene targeting and chromosome engineering.

  3. 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.

  4. 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.

  5. 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

  6. 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.

  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. PMID:27393468

  8. 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.

  9. Characterization of a conditional interleukin-1 receptor 1 mouse mutant using the Cre/LoxP system.

    PubMed

    Abdulaal, Wesam H; Walker, Catherine R; Costello, Ryan; Redondo-Castro, Elena; Mufazalov, Ilgiz A; Papaemmanouil, Athina; Rothwell, Nancy J; Allan, Stuart M; Waisman, Ari; Pinteaux, Emmanuel; Müller, Werner

    2016-04-01

    IL-1 is a key cytokine known to drive chronic inflammation and to regulate many physiological, immunological, and neuroimmunological responses via actions on diverse cell types of the body. To determine the mechanisms of IL-1 actions as part of the inflammatory response in vivo, we generated a conditional IL-1 receptor 1 (IL-1R1) mouse mutant using the Cre/LoxP system (IL-1R1(fl/fl) ). In the mutant generated, exon 5, which encodes part of the extracellular-binding region of the receptor, is flanked by LoxP sites, thereby inactivating the two previously described functional IL-1R1 gene transcripts after Cre-mediated recombination. Using keratin 14-Cre driver mice, new IL-1R1 deficient (-/-) mice were subsequently generated, in which all signaling IL-1 receptor isoforms are deleted ubiquitously. Furthermore, using vav-iCre driver mice, we deleted IL-1 receptor isoforms in the hematopoietic system. In these mice, we show that both the IL-17 and IL-22 cytokine response is reduced, when mice are challenged by the helminth Trichuris muris. We are currently crossing IL-1R1(fl/fl) mice with different Cre-expressing mice in order to study mechanisms of acute and chronic inflammatory diseases.

  10. Cardiomyocyte-specific deletion of leptin receptors causes lethal heart failure in Cre-recombinase-mediated cardiotoxicity.

    PubMed

    Hall, Michael E; Smith, Grant; Hall, John E; Stec, David E

    2012-12-15

    Although disruption of leptin signaling is associated with obesity as well as cardiac lipid accumulation and dysfunction, it has been difficult to separate the direct effects of leptin on the heart from those associated with the effects of leptin on body weight and fat mass. Using Cre-loxP recombinase technology, we developed tamoxifen-inducible, cardiomyocyte-specific leptin receptor-deficient mice to assess the role of leptin in regulating cardiac function. Cre recombinase activation in the heart resulted in transient reduction in left ventricular systolic function which recovered to normal levels by day 10. However, when cardiomyocyte leptin receptors were deleted in the setting of Cre recombinase-induced left ventricular dysfunction, irreversible lethal heart failure was observed in less than 10 days in all mice. Heart failure after leptin receptor deletion was associated with marked decreases of cardiac mitochondrial ATP, phosphorylated mammalian target of rapamycin (mTOR), and AMP-activated kinase (pAMPK). Our results demonstrate that specific deletion of cardiomyocyte leptin receptors, in the presence of increased Cre recombinase expression, causes lethal heart failure associated with decreased cardiac energy production. These observations indicate that leptin plays an important role in regulating cardiac function in the setting of cardiac stress caused by Cre-recombinase expression, likely through actions on cardiomyocyte energy metabolism.

  11. 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. PMID:18762802

  12. 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

  13. Generalized bacterial genome editing using mobile group II introns and Cre-lox

    PubMed Central

    Enyeart, Peter J; Chirieleison, Steven M; Dao, Mai N; Perutka, Jiri; Quandt, Erik M; Yao, Jun; Whitt, Jacob T; Keatinge-Clay, Adrian T; Lambowitz, Alan M; Ellington, Andrew D

    2013-01-01

    Efficient bacterial genetic engineering approaches with broad-host applicability are rare. We combine two systems, mobile group II introns (‘targetrons') and Cre/lox, which function efficiently in many different organisms, into a versatile platform we call GETR (Genome Editing via Targetrons and Recombinases). The introns deliver lox sites to specific genomic loci, enabling genomic manipulations. Efficiency is enhanced by adding flexibility to the RNA hairpins formed by the lox sites. We use the system for insertions, deletions, inversions, and one-step cut-and-paste operations. We demonstrate insertion of a 12-kb polyketide synthase operon into the lacZ gene of Escherichia coli, multiple simultaneous and sequential deletions of up to 120 kb in E. coli and Staphylococcus aureus, inversions of up to 1.2 Mb in E. coli and Bacillus subtilis, and one-step cut-and-pastes for translocating 120 kb of genomic sequence to a site 1.5 Mb away. We also demonstrate the simultaneous delivery of lox sites into multiple loci in the Shewanella oneidensis genome. No selectable markers need to be placed in the genome, and the efficiency of Cre-mediated manipulations typically approaches 100%. PMID:24002656

  14. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo

    PubMed Central

    Zariwala, Hatim A.; Borghuis, Bart G.; Hoogland, Tycho M.; Madisen, Linda; Tian, Lin; De Zeeuw, Chris I.; Zeng, Hongkui; Looger, Loren L.; Svoboda, Karel; Chen, Tsai-Wen

    2012-01-01

    Fluorescent calcium indicator proteins, such as GCaMP3, allow imaging of activity in genetically defined neuronal populations. GCaMP3 can be expressed using various gene delivery methods, such as viral infection or electroporation. However, these methods are invasive and provide inhomogeneous and non-stationary expression. Here we developed a genetic reporter mouse, Ai38, which expresses GCaMP3 in a Cre-dependent manner from the ROSA26 locus, driven by a strong CAG promoter. Crossing Ai38 with appropriate Cre mice produced robust GCaMP3 expression in defined cell populations in the retina, cortex and cerebellum. In the primary visual cortex, visually-evoked GCaMP3 signals showed normal orientation and direction selectivity. GCaMP3 signals were rapid, compared to virally expressed GCaMP3 and synthetic calcium indicators. In the retina, Ai38 allowed imaging spontaneous calcium waves in starburst amacrine cells during development, and light-evoked responses in ganglion cells in adult tissue. Our results show that the Ai38 reporter mouse provides a flexible method for targeted expression of GCaMP3. PMID:22378886

  15. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo.

    PubMed

    Zariwala, Hatim A; Borghuis, Bart G; Hoogland, Tycho M; Madisen, Linda; Tian, Lin; De Zeeuw, Chris I; Zeng, Hongkui; Looger, Loren L; Svoboda, Karel; Chen, Tsai-Wen

    2012-02-29

    Fluorescent calcium indicator proteins, such as GCaMP3, allow imaging of activity in genetically defined neuronal populations. GCaMP3 can be expressed using various gene delivery methods, such as viral infection or electroporation. However, these methods are invasive and provide inhomogeneous and nonstationary expression. Here, we developed a genetic reporter mouse, Ai38, which expresses GCaMP3 in a Cre-dependent manner from the ROSA26 locus, driven by a strong CAG promoter. Crossing Ai38 with appropriate Cre mice produced robust GCaMP3 expression in defined cell populations in the retina, cortex, and cerebellum. In the primary visual cortex, visually evoked GCaMP3 signals showed normal orientation and direction selectivity. GCaMP3 signals were rapid, compared with virally expressed GCaMP3 and synthetic calcium indicators. In the retina, Ai38 allowed imaging spontaneous calcium waves in starburst amacrine cells during development, and light-evoked responses in ganglion cells in adult tissue. Our results show that the Ai38 reporter mouse provides a flexible method for targeted expression of GCaMP3. PMID:22378886

  16. 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

  17. Non-specific protein–DNA interactions control I-CreI target binding and cleavage

    PubMed Central

    Molina, Rafael; Redondo, Pilar; Stella, Stefano; Marenchino, Marco; D’Abramo, Marco; Gervasio, Francesco Luigi; Charles Epinat, Jean; Valton, Julien; Grizot, Silvestre; Duchateau, Phillipe; Prieto, Jesús; Montoya, Guillermo

    2012-01-01

    Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis of protein–DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. Protein–DNA interaction engineering of homing endonucleases has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Protein–DNA interface studies have been focused mostly on specific contacts between amino acid side chains and bases to redesign the binding interface. However, it has been shown that 4 bp in the central DNA sequence of the 22-bp substrate of a homing endonuclease (I-CreI), which do not show specific protein–DNA interactions, is not devoid of content information. Here, we analyze the mechanism of target discrimination in this substrate region by the I-CreI protein, determining how it can occur independently of the specific protein–DNA interactions. Our data suggest the important role of indirect readout in this substrate region, opening the possibility for a fully rational search of new target sequences, thus improving the development of redesigned enzymes for therapeutic and biotechnological applications. PMID:22495931

  18. A transgenic Prox1-Cre-tdTomato reporter mouse for lymphatic vessel research.

    PubMed

    Bianchi, Roberta; Teijeira, Alvaro; Proulx, Steven T; Christiansen, Ailsa J; Seidel, Catharina D; Rülicke, Thomas; Mäkinen, Taija; Hägerling, René; Halin, Cornelia; Detmar, Michael

    2015-01-01

    The lymphatic vascular system plays an active role in immune cell trafficking, inflammation and cancer spread. In order to provide an in vivo tool to improve our understanding of lymphatic vessel function in physiological and pathological conditions, we generated and characterized a tdTomato reporter mouse and crossed it with a mouse line expressing Cre recombinase under the control of the lymphatic specific promoter Prox1 in an inducible fashion. We found that the tdTomato fluorescent signal recapitulates the expression pattern of Prox1 in lymphatic vessels and other known Prox1-expressing organs. Importantly, tdTomato co-localized with the lymphatic markers Prox1, LYVE-1 and podoplanin as assessed by whole-mount immunofluorescence and FACS analysis. The tdTomato reporter was brighter than a previously established red fluorescent reporter line. We confirmed the applicability of this animal model to intravital microscopy of dendritic cell migration into and within lymphatic vessels, and to fluorescence-activated single cell analysis of lymphatic endothelial cells. Additionally, we were able to describe the early morphological changes of the lymphatic vasculature upon induction of skin inflammation. The Prox1-Cre-tdTomato reporter mouse thus shows great potential for lymphatic research.

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

    PubMed Central

    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

    2015-01-01

    Recombinant adeno-associated viruses (rAAVs) are commonly used vehicles for in vivo gene transfer1-6. However, the tropism repertoire of naturally occurring AAVs is limited, prompting a search for novel AAV capsids with desired characteristics7-13. 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, AAV914-17, 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. PMID:26829320

  20. 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.

  1. Development of CRTEIL and CETRIZ, Cre-loxP-based systems, which allow change of expression of red to green or green to red fluorescence upon transfection with a cre-expression vector.

    PubMed

    Ohtsuka, Masato; Warita, Takayuki; Sakurai, Takayuki; Watanabe, Satoshi; Inoko, Hidetoshi; Sato, Masahiro

    2009-01-01

    We developed Cre-loxP-based systems, termed CRTEIL and CETRIZ, which allow gene switching in a noninvasive manner. Single transfection with pCRTEIL resulted in predominant expression of red fluorescence. Cotransfection with pCRTEIL and Cre-expression plasmid (pCAG/NCre) caused switching from red to green fluorescence. Similarly, cotransfection with pCETRIZ and pCAG/NCre resulted in change of green to red fluorescence. These noninvasive systems will be useful in cell lineage analysis, since descendants of cells exhibiting newly activated gene expression can be continuously monitored in noninvasive fashion.

  2. 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.

  3. 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.

  4. Col2CreERT2, A MOUSE MODEL FOR A CHONDROCYTE-SPECIFIC AND INDUCIBLE GENE DELETION

    PubMed Central

    Chen, M.; Li, S.; Xie, W.; Wang, B.; Chen, D.

    2014-01-01

    In 2007 and 2008, we published two articles reporting a tamoxifen (TM)-inducible, chondrocyte-specific gene-targeting mouse model in which the expression of CreERT2 is driven by the type II collagen promoter (Col2CreERT2). The fusion protein is specifically expressed and translocated into the nucleus upon TM administration, which in turn triggers gene recombination. Since then, this animal model has become a powerful tool to study the molecular mechanism of skeletal development and degenerative cartilage diseases, including knee joint osteoarthritis (OA), temporomandibular joint (TMJ) OA, and intervertebral disc (IVD) degeneration. In this review article, we summarise the application of Col2CreERT2 mice and discuss the potential usage of this animal model in a broad spectrum of cartilage development and molecular pathology studies. PMID:25340803

  5. Col2CreER(T2), a mouse model for a chondrocyte-specific and inducible gene deletion.

    PubMed

    Chen, M; Li, S; Xie, W; Wang, B; Chen, D

    2014-01-01

    In 2007 and 2008, we published two articles reporting a tamoxifen (TM)-inducible, chondrocyte-specific gene-targeting mouse model in which the expression of CreER(T2) is driven by the type II collagen promoter (Col2CreER(T2)). The fusion protein is specifically expressed and translocated into the nucleus upon TM administration, which in turn triggers gene recombination. Since then, this animal model has become a powerful tool to study the molecular mechanism of skeletal development and degenerative cartilage diseases, including knee joint osteoarthritis (OA), temporomandibular joint (TMJ) OA, and intervertebral disc (IVD) degeneration. In this review article, we summarise the application of Col2CreER(T2) mice and discuss the potential usage of this animal model in a broad spectrum of cartilage development and molecular pathology studies. PMID:25340803

  6. Development of selectable marker free, insect resistant, transgenic mustard (Brassica juncea) plants using Cre/lox mediated recombination

    PubMed Central

    2013-01-01

    Background Antibiotic/ herbicide resistant marker genes have been proven to be very useful in plant transformation for the initial selection of desired transgenic events. However, presence of these genes in the genetically modified crops may render the crop less acceptable to the consumers. Among several different approaches, the effectiveness of Cre/lox mediated recombination strategy for selectable marker gene (SMG) elimination has previously been demonstrated by different groups in several plants including Brassica. In the present study exploiting Cre/lox mediated recombination strategy, attempt has been made for selectable marker gene elimination from Allium sativum leaf agglutinin (ASAL) expressing Brassica plants with hemipteran insect resistant phenotype. Results Allium sativum leaf agglutinin (ASAL) linked with lox flanked hygromycin resistant (hpt) gene was introduced in mustard. Cre recombinase gene cassette was also integrated in separate event. A Cre/lox mediated recombination using crossing strategy was adopted to remove the hpt gene from the subsequent generation of selected hybrid events. Reciprocal crosses were made between T1ASAL-lox-hpt-lox and cre-bar plants. Marker gene elimination was confirmed in the resulting F1 hybrid progenies by PCR analysis, using hpt, cre and ASAL specific primers followed by Southern hybridization. In marker free plants, expression of ASAL was also confirmed by western blotting and ELISA analysis. Retention of functionality of expressed ASAL was investigated by agglutination assay using rabbit erythrocytes. Expressed ASAL was also found to be thermo-sensitive. In planta insect bioassay on F1 hybrid progenies exhibited detrimental effect on the performance of devastating target pest, Lipaphis erysimi. The F1 hybrid hpt negative, ASAL positive plants were allowed to self- fertilize to obtain F2 progeny plants. In some of these plants cre gene was found to be segregated out of the ASAL gene by genetic segregation yielding

  7. 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.

  8. Strategic quality planning.

    PubMed

    Yarborough, C M

    1994-04-01

    Instilling the principles of total quality management (TQM) into organizations is a difficult undertaking, yet many leaders of health care organizations are forging this paradigmatic shift with encouraging results. In this paper, a practical approach is described that has been deployed within an occupational and environmental health setting. The vehicle for this transformation is called the Strategic Quality Plan. Once the purpose was clearly elucidated by the leadership, a basic design of the plan was developed that melded two interrelated goals: achieving strategic business objectives by applying quality principles. The resulting matrix became the basis for detailed flowcharts that show the steps, inputs, decision points, deadlines, and continuous improvement loops for each strategic objective. A deployment plan was developed to aid implementation.

  9. 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.

  10. 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.

  11. 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.

  12. Homing endonuclease I-CreI derivatives with novel DNA target specificities

    PubMed Central

    Rosen, Laura E.; Morrison, Holly A.; Masri, Selma; Brown, Michael J.; Springstubb, Brendan; Sussman, Django; Stoddard, Barry L.; Seligman, Lenny M.

    2006-01-01

    Homing endonucleases are highly specific enzymes, capable of recognizing and cleaving unique DNA sequences in complex genomes. Since such DNA cleavage events can result in targeted allele-inactivation and/or allele-replacement in vivo, the ability to engineer homing endonucleases matched to specific DNA sequences of interest would enable powerful and precise genome manipulations. We have taken a step-wise genetic approach in analyzing individual homing endonuclease I-CreI protein/DNA contacts, and describe here novel interactions at four distinct target site positions. Crystal structures of two mutant endonucleases reveal the molecular interactions responsible for their altered DNA target specificities. We also combine novel contacts to create an endonuclease with the predicted target specificity. These studies provide important insights into engineering homing endonucleases with novel target specificities, as well as into the evolution of DNA recognition by this fascinating family of proteins. PMID:16971456

  13. Exploiting CReP for Knowledge Retrieval and Use in Complex Domains

    NASA Astrophysics Data System (ADS)

    Manenti, Lorenza; Sartori, Fabio

    Case Based Reasoning (CBR) is a Knowledge Management approach that consists in the development of decision support systems where problem are solved by analogy with similar problem solved in the past. In this way, the system supports users in finding solutions without starting from scratch. CBR has become a very important research topic in Artificial Intelligence, with the definition of methodologies and architectural patterns for supporting developers in the design and implementation of case-based systems. The paper presents one of this frameworks, namely CReP, an on-going research project of the Artificial Intelligence Laboratory (L.Int.Ar.) of University of Milan-Bicocca, focusing on the integration between CBR paradigm and metadata approach to obtain domain-independent case structure and retrieval algorithm definition.

  14. 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…

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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…

  1. 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.

  2. An open-source system for in planta gene stacking by Bxb1 and Cre recombinases.

    PubMed

    Hou, Lili; Yau, Yuan-Yeu; Wei, Junjie; Han, Zhiguo; Dong, Zhicheng; Ow, David W

    2014-12-01

    The rapid development of crops with multiple transgenic traits arouses the need for an efficient system for creating stacked cultivars. Most major crops rely on classical breeding to introgress the transgene from a laboratory variety to the numerous cultivars adapted to different growing regions. Even with vegetative propagated crops, genetic crosses are conducted during varietal improvement prior to vegetative cloning. The probability to assort the 'x' number of transgenic loci into a single genome may seem trivial, (¼) (x) for a diploid species, but given the 'y' number of other nontransgenic traits that breeders also need to assemble into the same genome, the (¼) (x+y) probability for a 'breeding stack' could quickly make the line conversion process unmanageable. Adding new transgenes onto existing transgenic varieties without creating a new segregating locus would require site-specific integration of new DNA at the existing transgenic locus. Here, we tested a recombinase-mediated gene-stacking scheme in tobacco. Sequential site-specific integration was mediated by the mycobacteriophage Bxb1 integrase-catalyzed recombination between attP and attB sites. Transgenic DNA no longer needed after integration was excised by Cre recombinase-mediated recombination of lox sites. Site-specific integration occurred in ~10% of the integration events, with half of those events usable as substrates for a next round of gene stacking. Among the site-specific integrants, however, a third experienced gene silencing. Overall, precise structure and reproducible expression of the sequentially added triple traits were obtained at an overall rate of ~3% of the transformed clones--a workable frequency for the development of commercial cultivars. Moreover, since neither the Bxb1-att nor the Cre-lox system is under patent, there is freedom to operate.

  3. Dysregulation of Uterine Signaling Pathways in Progesterone Receptor-Cre Knockout of Dicer

    PubMed Central

    Andreu-Vieyra, Claudia V.; Kim, Tae Hoon; Jeong, Jae-Wook; Hodgson, Myles C.; Chen, Ruihong; Creighton, Chad J.; Lydon, John P.; Gunaratne, Preethi H.; DeMayo, Francesco J.; Matzuk, Martin M.

    2012-01-01

    Epithelial-stromal interactions in the uterus are required for normal uterine functions such as pregnancy, and multiple signaling pathways are essential for this process. Although Dicer and microRNA (miRNA) have been implicated in several reproductive processes, the specific roles of Dicer and miRNA in uterine development are not known. To address the roles of miRNA in the regulation of key uterine pathways, we generated a conditional knockout of Dicer in the postnatal uterine epithelium and stroma using progesterone receptor-Cre. These Dicer conditional knockout females are sterile with small uteri, which demonstrate significant defects, including absence of glandular epithelium and enhanced stromal apoptosis, beginning at approximately postnatal d 15, with coincident expression of Cre and deletion of Dicer. Specific miRNA (miR-181c, −200b, −101, let-7d) were down-regulated and corresponding predicted proapoptotic target genes (Bcl2l11, Aldh1a3) were up-regulated, reflecting the apoptotic phenomenon. Although these mice had normal serum hormone levels, critical uterine signaling pathways, including progesterone-responsive genes, Indian hedgehog signaling, and the Wnt/β-catenin canonical pathway, were dysregulated at the mRNA level. Importantly, uterine stromal cell proliferation in response to progesterone was absent, whereas uterine epithelial cell proliferation in response to estradiol was maintained in adult uteri. These data implicate Dicer and appropriate miRNA expression as essential players in the regulation of multiple uterine signaling pathways required for uterine development and appropriate function. PMID:22798293

  4. 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…

  5. 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)

  6. Implementing College Strategic Plans.

    ERIC Educational Resources Information Center

    Further Education Development Agency, London (England).

    This document, which is intended for individuals responsible for planning and management at further education (FE) colleges, summarizes the factors that contributed to or obstructed implementation of strategic plans in 10 FE colleges in England and Wales in 1994-95. It presents key questions and illustrations that FE colleges can use for…

  7. A Strategic Planning Workbook.

    ERIC Educational Resources Information Center

    Austin, William

    This workbook outlines the Salem Community College's (New Jersey) Strategic Planning Initiative (SPI), which will enable the college to enter the 21st Century as an active agent in the educational advancement of the Salem community. SPI will allow college faculty, staff, students, and the local community to reflect on the vitality of the college…

  8. 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…

  9. Strategic Marketing for Presidents.

    ERIC Educational Resources Information Center

    Pappas, Richard J., Ed.

    Designed to inform the marketing efforts of community college presidents, this document describes the importance of marketing, presents a targeted approach, and outlines the specific roles and skills needed by the president to ensure successful efforts and effective institutions. The first chapter, "Developing a Marketing-Strategic Plan," by…

  10. 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.

  11. Strategic Goals for 2000.

    ERIC Educational Resources Information Center

    Hopper, Toni

    This is the report of the 2000 New Mexico Association of Community Colleges (NMACC) Strategic Planning Conference. Participants worked in small groups to identify the most probable and the most desirable world for the year 2000. Results for the most probable world include: (1) education would be available to many consumers, using many delivery…

  12. 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…

  13. Strategic Tutor Monitoring.

    ERIC Educational Resources Information Center

    Chee-kwong, Kenneth Chao

    1996-01-01

    Discusses effective tutor monitoring strategies based on experiences at the Open Learning Institute of Hong Kong. Highlights include key performance and strategic control points; situational factors, including tutor expectations and relevant culture; Theory X versus Theory Y leadership theories; and monitoring relationships with tutors. (LRW)

  14. 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…

  15. NAEA's Strategic Plans

    ERIC Educational Resources Information Center

    School Arts: The Art Education Magazine for Teachers, 2005

    2005-01-01

    This article discusses the National Art Education Association's strategic plan for art education. Their plan states that all PreK-12 students deserve a comprehensive, balanced, and sequential program of instruction in the visual arts. The Association's primary vision and goals are described.

  16. 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.

  17. 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)…

  18. 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.

  19. 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…

  20. The Strategic Revolution.

    PubMed

    Gardner, Andy

    2016-09-01

    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. PMID:27610556

  1. A novel bacterial artificial chromosome-transgenic podoplanin-cre mouse targets lymphoid organ stromal cells in vivo.

    PubMed

    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.

  2. Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice.

    PubMed

    Anastassiadis, Konstantinos; Fu, Jun; Patsch, Christoph; Hu, Shengbiao; Weidlich, Stefanie; Duerschke, Kristin; Buchholz, Frank; Edenhofer, Frank; Stewart, A Francis

    2009-01-01

    Tyrosine site-specific recombinases (SSRs) including Cre and FLP are essential tools for DNA and genome engineering. Cre has long been recognized as the best SSR for genome engineering, particularly in mice. Obtaining another SSR that is as good as Cre will be a valuable addition to the genomic toolbox. To this end, we have developed and validated reagents for the Dre-rox system. These include an Escherichia coli-inducible expression vector based on the temperature-sensitive pSC101 plasmid, a mammalian expression vector based on the CAGGs promoter, a rox-lacZ reporter embryonic stem (ES) cell line based on targeting at the Rosa26 locus, the accompanying Rosa26-rox reporter mouse line, and a CAGGs-Dre deleter mouse line. We also show that a Dre-progesterone receptor shows good ligand-responsive induction properties. Furthermore, we show that there is no crossover recombination between Cre-rox or Dre-loxP. Hence, we add another set of efficient tools to the genomic toolbox, which will enable the development of more sophisticated mouse models for the analysis of gene function and disease. PMID:19692579

  3. 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…

  4. 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…

  5. 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…

  6. Strategic market planning for hospitals.

    PubMed

    Zallocco, R L; Joseph, W B; Doremus, H

    1984-01-01

    The application of strategic market planning to hospital management is discussed, along with features of the strategic marketing management process. A portfolio analysis tool, the McKinsey/G.E. Business Screen, is presented and, using a large urban hospital as an example, discussed in detail relative to hospital administration. Finally, strategic implications of the portfolio analysis are examined.

  7. Developing a strategic security plan.

    PubMed

    Potter, Anthony N

    2011-01-01

    Strategic planning is an essential component of a security director job. Every healthcare organization has a strategic plan, and the security department that doesn't is at a major disadvantage when it comes to budgeting, growth and effectiveness. This article describes how a security director can develop and integrate a strategic plan into his organization's mission, vision and values.

  8. 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…

  9. Genetic and biochemical studies of poliovirus cis-acting replication element cre in relation to VPg uridylylation.

    PubMed

    Rieder, E; Paul, A V; Kim, D W; van Boom, J H; Wimmer, E

    2000-11-01

    In addition to highly conserved stem-loop structures located in the 5'- and 3'-nontranslated regions, genome replication of picornaviruses requires cis-acting RNA elements located in the coding region (termed cre) (K. L. McKnight and S. M. Lemon, J. Virol. 70:1941-1952, 1996; P. E. Lobert, N. Escriou, J. Ruelle, and T. Michiels, Proc. Natl. Acad. Sci. USA 96:11560-11565, 1999; I. Goodfellow, Y. Chaudhry, A. Richardson, J. Meredith, J. W. Almond, W. Barclay, and D. J. Evans, J. Virol. 74:4590-4600, 2000). cre elements appear to be essential for minus-strand RNA synthesis by an as-yet-unknown mechanism. We have discovered that the cre element of poliovirus (mapping to the 2C coding region of poliovirus type 1; nucleotides 4444 to 4505 in 2C), which is homologous to the cre element of poliovirus type 3, is preferentially used as a template for the in vitro uridylylation of VPg catalyzed by 3D(pol) in a reaction that is greatly stimulated by 3CD(pro) (A. V. Paul, E. Rieder, D. W. Kim, J. H. van Boom, and E. Wimmer, J. Virol. 74:10359-10370, 2000). Here we report a direct correlation between mutations that eliminate, or severely reduce, the in vitro VPg-uridylylation reaction and produce replication phenotypes in vivo. None of the genetic changes significantly influenced translation or polyprotein processing. A substitution mapping to the first A (A4472C) of a conserved AAACA sequence in the loop of PV-cre(2C) eliminated the ability of the cre RNA to serve as template for VPg uridylylation and abolished RNA infectivity. Mutagenesis of the second A (A4473C; AAACA) severely reduced the yield of VPgpUpU and RNA infectivity was restored only after reversion to the wild-type sequence. The effect of substitution of the third A (A4474G; AAACA) was less severe but reduced both VPg uridylylation and virus yield. Disruption of base pairing within the upper stem region of PV-cre(2C) also affected uridylylation of VPg. Virus derived from transcripts containing mutations in the stem

  10. Preventive Health Behaviors Among Grandmothers Raising Grandchildren

    PubMed Central

    Silverstein, Merril

    2009-01-01

    Objectives We examined differential preventive health behavior among grandmothers who recently began raising a grandchild, grandmothers raising a grandchild for at least 2 years, and grandmothers not raising a grandchild. Methods Data came from the 2000, 2002, and 2004 waves of the Health and Retirement Study. We ran multivariate logistic regression models to assess receipt of influenza vaccination, cholesterol screening, monthly breast self-exam, mammography, and Papanicolaou (Pap) tests among grandmothers aged 50 to 75. Results Grandmothers who recently began raising a grandchild were significantly less likely to report influenza vaccination and cholesterol screening than grandmothers not raising grandchildren, even after we controlled for increased emotional and financial strains within the household. We also observed this association for Pap tests, although this finding was only marginally significant. Grandmothers who had been raising a grandchild for at least 2 years were significantly more likely to report influenza vaccination and monthly breast self-exam than grandmothers not raising grandchildren. Discussion The enhancement of preventive behavior seen among long-term grandparent caregivers does not fully offset the suppression of preventive behavior during the transition into care; support groups should target a range of interventions toward the promotion of healthy behavior among new grandparent caregivers. PMID:18818451

  11. A Cre-conditional MYCN-driven neuroblastoma mouse model as an improved tool for preclinical studies.

    PubMed

    Althoff, K; Beckers, A; Bell, E; Nortmeyer, M; Thor, T; Sprüssel, A; Lindner, S; De Preter, K; Florin, A; Heukamp, L C; Klein-Hitpass, L; Astrahantseff, K; Kumps, C; Speleman, F; Eggert, A; Westermann, F; Schramm, A; Schulte, J H

    2015-06-01

    Neuroblastoma, a childhood cancer that originates from neural crest-derived cells, is the most common deadly solid tumor of infancy. Amplification of the MYCN oncogene, which occurs in approximately 20-25% of human neuroblastomas, is the most prominent genetic marker of high-stage disease. The availability of valid preclinical in vivo models is a prerequisite to develop novel targeted therapies. We here report on the generation of transgenic mice with Cre-conditional induction of MYCN in dopamine β-hydroxylase-expressing cells, termed LSL-MYCN;Dbh-iCre. These mice develop neuroblastic tumors with an incidence of >75%, regardless of strain background. Molecular profiling of tumors revealed upregulation of the MYCN-dependent miR-17-92 cluster as well as expression of neuroblastoma marker genes, including tyrosine hydroxylase and the neural cell adhesion molecule 1. Gene set enrichment analyses demonstrated significant correlation with MYC-associated expression patterns. Array comparative genome hybridization showed that chromosomal aberrations in LSL-MYCN;Dbh-iCre tumors were syntenic to those observed in human neuroblastomas. Treatment of a cell line established from a tumor derived from a LSL-MYCN;Dbh-iCre mouse with JQ1 or MLN8237 reduced cell viability and demonstrated oncogene addiction to MYCN. Here we report establishment of the first Cre-conditional human MYCN-driven mouse model for neuroblastoma that closely recapitulates the human disease with respect to tumor localization, histology, marker expression and genomic make up. This mouse model is a valuable tool for further functional studies and to assess the effect of targeted therapies. PMID:25174395

  12. Metabolic and Behavioural Phenotypes in Nestin-Cre Mice Are Caused by Hypothalamic Expression of Human Growth Hormone

    PubMed Central

    Pruniau, Vincent P. E. G.; Stijnen, Pieter; de Faudeur, Geoffroy; Tuand, Krizia; Meulemans, Sandra; Serneels, Lutgarde; Schraenen, Anica; Schuit, Frans; Creemers, John W. M.

    2015-01-01

    The Nestin-Cre driver mouse line has mild hypopituitarism, reduced body weight, a metabolic phenotype and reduced anxiety. Although several causes have been suggested, a comprehensive explanation is still lacking. In this study we examined the molecular mechanisms leading to this compound phenotype. Upon generation of the Nestin-Cre mice, the human growth hormone (hGH) minigene was inserted downstream of the Cre recombinase to ensure efficient transgene expression. As a result, hGH is expressed in the hypothalamus. This results in the auto/paracrine activation of the GH receptor as demonstrated by the increased phosphorylation of signal transducer and activator of transcription 5 (STAT5) and reduced expression of growth hormone releasing hormone (Ghrh). Low Ghrh levels cause hypopituitarism consistent with the observed mouse growth hormone (mGH) deficiency. mGH deficiency caused reduced activation of the GH receptor and hence reduced phosphorylation of STAT5 in the liver. This led to decreased levels of hepatic Igf-1 mRNA and consequently postnatal growth retardation. Furthermore, genes involved in lipid uptake and synthesis, such as CD36 and very low-density lipoprotein receptor were upregulated, resulting in liver steatosis. In conclusion, this study demonstrates the unexpected expression of hGH in the hypothalamus of Nestin-Cre mice which is able to activate both the GH receptor and the prolactin receptor. Increased hypothalamic GH receptor signaling explains the observed hypopituitarism, reduced growth and metabolic phenotype of Nestin-Cre mice. Activation of either receptor is consistent with reduced anxiety. PMID:26275221

  13. Fund Raising. The Guide to Raising Money from Private Sources. Second Edition, Revised and Enlarged.

    ERIC Educational Resources Information Center

    Broce, Thomas E.

    A basic guide and reference to raising funds from private sources is presented that is designed primarily for professional personnel or the administrator employing fund-raising personnel. The first edition of the guide has also been used as a textbook for university courses in fund-raising. This revised edition includes new materials on: capital…

  14. Common Surgeries Raise Risk for Opioid Dependence

    MedlinePlus

    ... fullstory_159815.html Common Surgeries Raise Risk for Opioid Dependence: Study Doctors should explore alternatives for pain ... have an elevated risk of growing dependent on opioid painkillers, a new study finds. These prescription painkillers ...

  15. 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)

  16. Pain Raises Risk of Opioid Addiction

    MedlinePlus

    ... fullstory_160033.html Pain Raises Risk of Opioid Addiction Men and younger people had higher odds of ... had a 41 percent higher risk of opioid addiction than those with no pain. That increased risk ...

  17. Alcohol, Obesity Could Raise Esophageal Cancer Risk

    MedlinePlus

    ... page: https://medlineplus.gov/news/fullstory_160133.html Alcohol, Obesity Could Raise Esophageal Cancer Risk A third ... now linked to 11 types of cancer and alcohol links to six," she said in an institute ...

  18. Raised replacement heifers: some economic considerations.

    PubMed

    Hughes, Harlan

    2013-11-01

    This article lays out the recommended procedure for determining the economic cost of developing a proposed set of replacement heifers. In particular, a 6-step process that calculates the economic cost of developing a raised pregnant replacement heifer is described.

  19. 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.

  20. The Strategic Petroleum Reserve

    SciTech Connect

    Not Available

    1991-01-01

    The Strategic Petroleum Reserve program was set into motion by the 1975 Energy Policy and Conservation Act (EPCA). By 1990, 590 million barrels of oil had been placed in storage. Salt domes along the Gulf Coast offered ideal storage. Both sweet'' and sour'' crude oil have been acquired using various purchase options. Drawdown, sale, and distribution of the oil would proceed according to guidelines set by EPCA in the event of a severe energy supply disruption. (SM)

  1. 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.

  2. Strategic scene generation model

    NASA Astrophysics Data System (ADS)

    Heckathorn, Harry M.; Anding, David C.

    1992-09-01

    The Strategic Defense Initiative (SDI) must simulate the detection, acquisition, discrimination and tracking of anticipated targets and predict the effect of natural and man-made background phenomena on optical sensor systems designed to perform these tasks. NRL is developing such a capability using a computerized methodology to provide modeled data in the form of digital realizations of complex, dynamic scenes. The Strategic Scene Generation Model (SSGM) is designed to integrate state-of-science knowledge, data bases and computerized phenomenology models to simulate strategic engagement scenarios and to support the design, development and test of advanced surveillance systems. Multi-phenomenology scenes are produced from validated codes--thereby serving as a standard against which different SDI concepts and designs can be tested. This paper describes the SSGM design architecture, the software modules and databases which are used to create scene elements, the synthesis of deterministic and/or stochastic structured scene elements into composite scenes, the software system to manage the various databases and digital image libraries, and verification and validation by comparison with measured data. The focus will be on the functionality and development schedule of the Baseline Model (SSGMB) which is currently being implemented.

  3. Second-generation Notch1 activity-trap mouse line (N1IP::CreHI) provides a more comprehensive map of cells experiencing Notch1 activity.

    PubMed

    Liu, Zhenyi; Brunskill, Eric; Boyle, Scott; Chen, Shuang; Turkoz, Mustafa; Guo, Yuxuan; Grant, Rachel; Kopan, Raphael

    2015-03-15

    We have previously described the creation and analysis of a Notch1 activity-trap mouse line, Notch1 intramembrane proteolysis-Cre6MT or N1IP::Cre(LO), that marked cells experiencing relatively high levels of Notch1 activation. Here, we report and characterize a second line with improved sensitivity (N1IP::Cre(HI)) to mark cells experiencing lower levels of Notch1 activation. This improvement was achieved by increasing transcript stability and by restoring the native carboxy terminus of Cre, resulting in a five- to tenfold increase in Cre activity. The magnitude of this effect probably impacts Cre activity in strains with carboxy-terminal Ert2 fusion. These two trap lines and the related line N1IP::Cre(ERT2) form a complementary mapping tool kit to identify changes in Notch1 activation patterns in vivo as the consequence of genetic or pharmaceutical intervention, and illustrate the variation in Notch1 signal strength from one tissue to the next and across developmental time. PMID:25725069

  4. Second-generation Notch1 activity-trap mouse line (N1IP::CreHI) provides a more comprehensive map of cells experiencing Notch1 activity

    PubMed Central

    Liu, Zhenyi; Brunskill, Eric; Boyle, Scott; Chen, Shuang; Turkoz, Mustafa; Guo, Yuxuan; Grant, Rachel; Kopan, Raphael

    2015-01-01

    We have previously described the creation and analysis of a Notch1 activity-trap mouse line, Notch1 intramembrane proteolysis-Cre6MT or N1IP::CreLO, that marked cells experiencing relatively high levels of Notch1 activation. Here, we report and characterize a second line with improved sensitivity (N1IP::CreHI) to mark cells experiencing lower levels of Notch1 activation. This improvement was achieved by increasing transcript stability and by restoring the native carboxy terminus of Cre, resulting in a five- to tenfold increase in Cre activity. The magnitude of this effect probably impacts Cre activity in strains with carboxy-terminal Ert2 fusion. These two trap lines and the related line N1IP::CreERT2 form a complementary mapping tool kit to identify changes in Notch1 activation patterns in vivo as the consequence of genetic or pharmaceutical intervention, and illustrate the variation in Notch1 signal strength from one tissue to the next and across developmental time. PMID:25725069

  5. Mx1-Cre mediated Rgs12 conditional knockout mice exhibit increased bone mass phenotype

    PubMed Central

    Yang, Shuying; Li, Yi-ping; Liu, Tongjun; He, Xiaoning; Yuan, Xue; Li, Chunyi; Cao, Jay; Kim, Yunjung

    2013-01-01

    Regulators of G-protein Signaling (Rgs) proteins are the members of a multigene family of GTPase-accelerating proteins (GAP) for the Galpha subunit of heterotrimeric G-proteins. Rgs proteins play critical roles in the regulation of G protein couple receptor (GPCR) signaling in normal physiology and human diseases such as cancer, heart diseases and inflammation. Rgs12 is the largest protein of the Rgs protein family. Some in vitro studies have demonstrated that Rgs12 plays a critical role in regulating cell differentiation and migration; however its function and mechanism in vivo is largely unknown. Here, we generated a floxed Rgs12 allele (Rgs12flox/flox) in which the exon 2, containing both PDZ and PTB_PID domains of Rgs12, was flanked with two loxp sites. By using the inducible Mx1-cre and Poly I:C system to specifically delete Rgs12 at postnatal 10 days in interferon-responsive cells including monocyte and macrophage cells, we found that Rgs12 mutant mice had growth retardation with the phenotype of increased bone mass. We further found that deletion of Rgs12 reduced osteoclast numbers and had no significant effect on osteoblast formation. Thus, Rgs12flox/flox conditional mice provide a valuable tool for in vivo analysis of Rgs12 function and mechanism through time- and cell-specific deletion of Rgs12. PMID:23349096

  6. 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

  7. Camk2a-Cre-Mediated Conditional Deletion of Chromatin Remodeler Brg1 Causes Perinatal Hydrocephalus

    PubMed Central

    Cao, Mou; Wu, Jiang I.

    2015-01-01

    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/SmarcA4or 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 deletionin 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 narrowerSylvian 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. PMID:25929186

  8. FLP/FRT and Cre/lox recombination technology in C. elegans

    PubMed Central

    2014-01-01

    One of the most powerful aspects of biological inquiry using model organisms is the ability to control gene expression. A holy grail is both temporal and spatial control of the expression of specific gene products – that is, the ability to express or withhold the activity of genes or their products in specific cells at specific times. Ideally such a method would also regulate the precise levels of gene activity, and alterations would be reversible. The related goal of controlled or purposefully randomized expression of visible markers is also tremendously powerful. While not all of these feats have been accomplished in C. elegans to date, much progress has been made, and recent technologies put these goals within closer reach. Here, I present published examples of successful two-component site-specific recombination in C. elegans. These technologies are based on the principle of controlled intra-molecular excision or inversion of DNA sequences between defined sites, as driven by FLP or Cre recombinases. I discuss several prospects for future applications of this technology. PMID:24874786

  9. 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.

  10. Flow cytometry and single nucleus sorting for Cre-based analysis of changes in transcriptional states.

    PubMed

    Samadder, Partha; Weng, Ning; Doetschman, Thomas; Heimark, Ronald L; Galbraith, David W

    2016-05-01

    The organs of eukaryotic organisms comprise complex interspersions of cell types, whose different molecular activities, and corresponding cellular states, cooperate during development to produce the final, functional organ. Dysfunction of organs in disease, particularly oncogenesis, initiates with changes of state of a minor subset of cells. It therefore is hard to detect early molecular indicators of disease within an overwhelming background of normal cells. Flow cytometry and sorting provides a convenient way to purify minority subpopulations, if a specific fluorophore can be unambiguously and exclusively associated with this subpopulation. We have generated a number of transgenic mouse lines expressing a nuclear-localized version of the Green Fluorescent Protein (GFP), within which the production of a chimeric histone 2B-GFP protein occurs under the control of a constitutively-active, actin-derived promoter, separated by a Floxed-STOP sequence. In the presence of Cre recombinase, within F1 progeny of these mouse lines, excision of the STOP sequence activates transcription which results in the emergence of cells containing green fluorescent nuclei. We describe the characterization of these lines using a combination of microscopic imaging, flow cytometry and sorting, and Reverse-Transcription polymerase chain reaction of transcripts within single sorted nuclei isolated from tissue homogenates. These lines should be particularly useful for analysis of transcriptional changes in oncogenesis. © 2016 International Society for Advancement of Cytometry. PMID:27003621

  11. Tamoxifen-Dependent, Inducible Bmp2CreER Drives Selective Recombinase Activity in Early Interdigital Mesenchyme and Digit Collateral Ligaments

    PubMed Central

    Huang, Bau-Lin; Mackem, Susan

    2015-01-01

    During limb development, the interdigital mesenchyme has been proposed to play a signaling role instructing morphogenesis of different digit types, as well as undergoing programmed cell death necessary to free digits in animals not adapted for swimming or flying. We have generated a conditional, tamoxifen-dependent Cre line, Bmp2CreER, which drives highly selective recombination restricted to the distal limb mesoderm, largely restricted to the interdigits, and selectively active in digit ligament but not tendon progenitors at later stages. The Bmp2CreER provides a valuable new tool to dissect roles of interdigital mesenchyme and potentially investigate divergence of ligament and tendon lineages. PMID:25850076

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

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... 50 Wildlife and Fisheries 1 2010-10-01 2010-10-01 false Live farm-raised fish and farm-raised fish eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

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

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... 50 Wildlife and Fisheries 1 2014-10-01 2014-10-01 false Live farm-raised fish and farm-raised fish eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

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

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... 50 Wildlife and Fisheries 1 2012-10-01 2012-10-01 false Live farm-raised fish and farm-raised fish eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

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

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... 50 Wildlife and Fisheries 1 2011-10-01 2011-10-01 false Live farm-raised fish and farm-raised fish eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

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

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... 50 Wildlife and Fisheries 1 2013-10-01 2013-10-01 false Live farm-raised fish and farm-raised fish eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  17. 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.

  18. Raising venture capital in the biopharma industry.

    PubMed

    Leytes, Lev J

    2002-11-15

    Raising venture capital (VC) is both an art and a science. Future entrepreneurs should carefully consider the various issues of VC financing that have a strong impact on the success of their business. In addition to attracting the best venture capital firms, these issues include such subtle but important points as the timing of financing (especially of the first round), external support sources, desirable qualities of a VC firm, amount to be raised, establishing a productive interface between the founders and the venture capitalists, and most importantly the effects of well-executed VC funding on hiring senior executives and scientific leaders.

  19. Raising a gender non-conforming child.

    PubMed

    Wisnowski, Deborah L

    2011-10-01

    This article overviews the challenges parents face raising gender non-conforming children. The author is the mother of a fifteen year old gender non-conforming (GNC) child and co-founder of Stepping Stones Support Group which organizes support programs for families of GNC. The article discusses social challenges, educational challenges and internal conflicts the author has experienced while raising her child. The author also discusses the process of founding Stepping Stones and the importance having support has played in her and her child's life.

  20. The strategic use of humility.

    PubMed

    Kerfoot, K

    1998-01-01

    Authentic humility is strategic for the success of leaders. It is strategic from the perspective of the egotistically free relationship humility establishes with the staff, patients, and others. The lack of strategic humility explains the inevitable downfall of people who are motivated to lead by power and status. It also explains why leaders who always remember where they came from, and who are motivated by what they can do for patients and staff, succeed.

  1. 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.

  2. 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.

  3. 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.

  4. A Tmprss2-CreERT2 Knock-In Mouse Model for Cancer Genetic Studies on Prostate and Colon.

    PubMed

    Gao, Dong; Zhan, Yu; Di, Wei; Moore, Amanda R; Sher, Jessica J; Guan, Youxin; Wang, Shangqian; Zhang, Zeda; Murphy, Devan A; Sawyers, Charles L; Chi, Ping; Chen, Yu

    2016-01-01

    Fusion between TMPRSS2 and ERG, placing ERG under the control of the TMPRSS2 promoter, is the most frequent genetic alteration in prostate cancer, present in 40-50% of cases. The fusion event is an early, if not initiating, event in prostate cancer, implicating the TMPRSS2-positive prostate epithelial cell as the cancer cell of origin in fusion-positive prostate cancer. To introduce genetic alterations into Tmprss2-positive cells in mice in a temporal-specific manner, we generated a Tmprss2-CreERT2 knock-in mouse. We found robust tamoxifen-dependent Cre activation in the prostate luminal cells but not basal epithelial cells, as well as epithelial cells of the bladder and gastrointestinal (GI) tract. The knock-in allele on the Tmprss2 locus does not noticeably impact prostate, bladder, or gastrointestinal function. Deletion of Pten in Tmprss2-positive cells of adult mice generated neoplasia only in the prostate, while deletion of Apc in these cells generated neoplasia only in the GI tract. These results suggest that this new Tmprss2-CreERT2 mouse model will be a useful resource for genetic studies on prostate and colon. PMID:27536883

  5. 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.

  6. 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.

  7. A Tmprss2-CreERT2 Knock-In Mouse Model for Cancer Genetic Studies on Prostate and Colon

    PubMed Central

    Zhan, Yu; Di, Wei; Moore, Amanda R.; Sher, Jessica J.; Guan, Youxin; Wang, Shangqian; Zhang, Zeda; Murphy, Devan A.; Sawyers, Charles L.; Chi, Ping; Chen, Yu

    2016-01-01

    Fusion between TMPRSS2 and ERG, placing ERG under the control of the TMPRSS2 promoter, is the most frequent genetic alteration in prostate cancer, present in 40–50% of cases. The fusion event is an early, if not initiating, event in prostate cancer, implicating the TMPRSS2-positive prostate epithelial cell as the cancer cell of origin in fusion-positive prostate cancer. To introduce genetic alterations into Tmprss2-positive cells in mice in a temporal-specific manner, we generated a Tmprss2-CreERT2 knock-in mouse. We found robust tamoxifen-dependent Cre activation in the prostate luminal cells but not basal epithelial cells, as well as epithelial cells of the bladder and gastrointestinal (GI) tract. The knock-in allele on the Tmprss2 locus does not noticeably impact prostate, bladder, or gastrointestinal function. Deletion of Pten in Tmprss2-positive cells of adult mice generated neoplasia only in the prostate, while deletion of Apc in these cells generated neoplasia only in the GI tract. These results suggest that this new Tmprss2-CreERT2 mouse model will be a useful resource for genetic studies on prostate and colon. PMID:27536883

  8. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 1 2010-01-01 2010-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture... Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the application must be developed in accordance with the following four key principles: (1) Strategic vision...

  9. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 1 2014-01-01 2014-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture... Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the... transportation, education, and public safety. (b) Minimum requirements. The strategic plan must: (1) Describe...

  10. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 1 2012-01-01 2012-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture... Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the... transportation, education, and public safety. (b) Minimum requirements. The strategic plan must: (1) Describe...

  11. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 1 2013-01-01 2013-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture... Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the... transportation, education, and public safety. (b) Minimum requirements. The strategic plan must: (1) Describe...

  12. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 1 2011-01-01 2011-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture... Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the... transportation, education, and public safety. (b) Minimum requirements. The strategic plan must: (1) Describe...

  13. Grammatical Consciousness-Raising and the Lexicon.

    ERIC Educational Resources Information Center

    Bland, Susan Kesner

    A role is proposed for the lexicon in the grammatical consciousness-raising (GCR) framework for second language teaching outlined by W. Rutherford and M. Sharwood Smith (1987, 1988). First, a lexical framework that offers a series of questions to help teachers discover the relevant semantic and pragmatic information associated with grammatical…

  14. Grandparents Raising Grandchildren: Help from Cooperative Extension

    ERIC Educational Resources Information Center

    Crocoll, Caroline E.

    2004-01-01

    Grandparents raising grandchildren face many challenges including dealing with the complicated systems of child welfare, education, healthcare, and the aging network. They must contend with maintaining financial resources for their grandchildren, enrolling the children in school, accessing healthcare and insurance, and obtaining safe, affordable…

  15. 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…

  16. Infant Botulism and Raised Intraocular Pressure.

    PubMed

    Eberly, Matthew D; Uber, Ian; Kieling, Christopher R; Birdsong, Richard H

    2009-11-01

    Infant botulism is an exceedingly rare disease. Because confirmatory laboratory testing is not available for several days after time of presentation, infant botulism remains a clinical diagnosis. The authors demonstrate how raised intraocular pressure may provide an additional clinical clue to making the diagnosis.

  17. Raising Minority Achievement in Science and Math.

    ERIC Educational Resources Information Center

    Hrabowski, Freeman A., III

    2003-01-01

    Describes the achievement gap between minority and non-minority students in science and mathematics. Provides advice to parents, educators, and students on how to raise minority-group achievement. Describes the Meyerhoff Scholars Program at the University of Maryland, Baltimore County, which recruits and supports minority students who excel in…

  18. 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 Community?" (Martin Luther…

  19. 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…

  20. 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…

  1. 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…

  2. Fund Raising in ARL Libraries. Kit 94.

    ERIC Educational Resources Information Center

    Association of Research Libraries, Washington, DC. Office of Management Studies.

    In January 1983 the Association of Research Libraries (ARL) surveyed its members regarding their fund raising programs. A response rate of 85% (95 out of 113 libraries surveyed) was obtained. It was found that 85% of ARL member libraries had been involved in university-wide fund campaigns; that over 75% of the respondents had also conducted…

  3. 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 Foundation…

  4. 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…

  5. Raising Academic Achievement through Parent Involvement.

    ERIC Educational Resources Information Center

    Blackfelner, Carol; Ranallo, Barbara

    Research has demonstrated that parent involvement has many beneficial effects for students. This action research project designed and implemented a program to raise the academic achievement of second-grade students by increasing parent involvement. The students attended two second-grade classrooms in a west-central Illinois school. The problem of…

  6. 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…

  7. 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…

  8. 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…

  9. Parents Matter: Tips for Raising Teenagers.

    ERIC Educational Resources Information Center

    Campaign for Tobacco-Free Kids, Washington, DC.

    It is easy for parents to feel that they have lost their influence over their children once children reach adolescence. Several organizations with different focuses but with the same goal of helping parents raise their teenagers banded together to develop this publication. Their message is that parents can do many things to foster their children's…

  10. 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…

  11. 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.

  12. Strategic identities in cyberspace.

    PubMed

    Talamo, A; Ligorio, B

    2001-02-01

    This paper aims at describing, according to the recent advances in social psychology and Computer Mediated Communication, how identities are perceived and constructed in cyberspace. All interactions analyzed in this study were performed within "Euroland," a collaborative virtual environment. The interacting community was composed of students, teachers, and researchers working on a transnational educational project. Practices and dialogues within Euroland are analyzed using an ethnographic and conversational method. A sample of discourses and actions that occurred during 8 months of time, selected according to the research aims, was analyzed. During online connections, users were personified by an "Avatar." Avatars are able to walk, fly, and look around the virtual world. They are also able to build and manipulate three-dimensional objects, perform virtual actions, and chat with other connected users. Results showed that "Eurolanders" showed and constructed their identities using strategic "positioning" depending on the interactive situation. Identities are thus dynamic and strongly related to the context, created and constantly recreated by the users. It is concluded that specific features offered by the Euroland environment are exploited by the users as resources to play with, while moving from one strategic positioning to another. Cyber identities involve resources given by specific technological tools and by community. The cyber-identity construction process seems to be highly congruent to the advances in the dialogical perspective in psychology, where identities are considered in their conceptualizations as multiple, "multivoiced," "positioned," and context-dependent.

  13. 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

  14. Strategic Decisions in Task-Oriented Reading.

    PubMed

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

    2015-01-01

    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.

  15. Sox2-CreER mice are useful for fate mapping of mature, but not neonatal, cochlear supporting cells in hair cell regeneration studies

    PubMed Central

    Walters, Bradley J.; Yamashita, Tetsuji; Zuo, Jian

    2015-01-01

    Studies of hair cell regeneration in the postnatal cochlea rely on fate mapping of supporting cells. Here we characterized a Sox2-CreER knock-in mouse line with two independent reporter mouse strains at neonatal and mature ages. Regardless of induction age, reporter expression was robust, with CreER activity being readily detectable in >85% of supporting cells within the organ of Corti. When induced at postnatal day (P) 28, Sox2-CreER activity was exclusive to supporting cells demonstrating its utility for fate mapping studies beyond this age. However, when induced at P1, Sox2-CreER activity was also detected in >50% of cochlear hair cells, suggesting that Sox2-CreER may not be useful to fate map a supporting cell origin of regenerated hair cells if induced at neonatal ages. Given that this model is currently in use by several investigators for fate mapping purposes, and may be adopted by others in the future, our finding that current protocols are effective for restricting CreER activity to supporting cells at mature but not neonatal ages is both significant and timely. PMID:26108463

  16. 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…

  17. The Search for Strategic Silence.

    ERIC Educational Resources Information Center

    Lentz, Richard

    Media content analysts seldom observe the principle that editorial omissions are as telling as what is published or broadcast; hence, the purpose of this paper is to explore, and thus stimulate debate about, editorial omissions or "strategic silence." It is observed that as a concept, strategic silence embraces both tact and strategy--the former…

  18. 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…

  19. 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…

  20. 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.

  1. Strategic Planning for Curriculum Reform.

    ERIC Educational Resources Information Center

    Cawelti, Gordon

    Strategic planning is a process deliberately designed to help educational leaders conceive of the kinds of institutions that they would like to create to serve their students. Drawn from industry, strategic planning attempts to respond to a wide variety of factors determining the directions that schools should take in planning for the future.…

  2. Strategic Marketing for Educational Systems.

    ERIC Educational Resources Information Center

    Hanson, E. Mark; Henry, Walter

    1992-01-01

    Private-sector strategic marketing processes can significantly benefit schools desiring to develop public confidence and support and establish guidelines for future development. This article defines a strategic marketing model for school systems and articulates the sequence of related research and operational steps comprising it. Although schools…

  3. 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)

  4. 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 sustainable…

  5. Potential effects of raising Medicare's eligibility age.

    PubMed

    Waidmann, T A

    1998-01-01

    Recent fiscal pressures on Medicare and an already enacted increase in Social Security's normal retirement age have generated discussion of raising Medicare's age of entitlement. This DataWatch examines potential impacts of raising Medicare's eligibility age to sixty-seven on public-sector health spending and individual insurance coverage. The proposed increase would affect a substantial fraction of beneficiaries without having a commensurate effect on expenditures, even in the long run. It is estimated that if the eligibility age were sixty-seven, upwards of 500,000 persons ages sixty-five and sixty-six would be left without any insurance, and even more would not be able to afford coverage with benefits similar to those of Medicare. PMID:9558794

  6. 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.

  7. 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…

  8. Generation of a transgenic mouse for colorectal cancer research with intestinal cre expression limited to the large intestine.

    PubMed

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

    2010-08-01

    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 used 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 APC(580S) mice to generate mice with APC inactivation at one (CAC;APC(580S/+)) or both alleles (CAC;APC(580S/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-week-old CAC;APC(580S/+) mice, but CAC;APC(580S/580S) mice had significant mucosal hyperplasia in the colon at 3 weeks, which developed into tumors by 6 weeks. By 10 weeks, 20% of CAC;APC(580S/+) 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, that is, transgene expression is limited to the epithelium of the large bowel with normal cells found next to genetically modified cells.

  9. 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.

  10. 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.

  11. Inducible activation of Cre recombinase in adult mice causes gastric epithelial atrophy, metaplasia, and regenerative changes in the absence of “floxed” alleles

    PubMed Central

    Jae Huh, Won; Mysorekar, Indira U.

    2010-01-01

    The epithelium of the mammalian gastric body comprises multiple cell types replenished by a single stem cell. The adult conformation of cell lineages occurs well after birth; hence, study of genes regulating stem cell activity is facilitated by inducible systems for gene deletion. However, there is a potential pitfall involving the commonly used inducible Cre recombinase system to delete genes: we report here that induction of Cre using standard doses of tamoxifen led to marked spasmolytic polypeptide-expressing metaplasia of the stomach within days and profound atrophy of the entire epithelium with foci of hyperplasia by 2 wk even in the absence of loxP-flanked alleles. Cre induction caused genotoxicity with TdT-mediated dUTP nick-end labeling (TUNEL)-positive apoptosis (TUNEL-positive cells) and increased levels of DNA damage markers (γH2AX, p53, DDIT3, GADD45A). Although Cre was expressed globally by use of a chicken actin promoter, the effects were almost entirely stomach specific. Despite severe injury, a subset of mice showed near complete healing of the gastric mucosa 11–12 wk after Cre induction, suggesting substantial gastric regenerative capacity. Finally, we show that nongenotoxic doses of tamoxifen could be used to specifically delete loxP-flanked Bmpr1a, the receptor for bone morphogenetic protein 2, 4, and 7, causing antral polyps and marked antral-pyloric hyperplasia, consistent with previous reports on Bmpr1a. Together, the results show dose-dependent, potentially reversible sensitivity of the gastric mucosa to Cre genotoxicity. Thus we propose that tamoxifen induction of Cre could be used to induce genotoxic injury to study the regenerative capacity of the gastric epithelial stem cell. PMID:20413717

  12. Endogenous n-3 polyunsaturated fatty acids delay progression of pancreatic ductal adenocarcinoma in Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) mice.

    PubMed

    Mohammed, Altaf; Janakiram, Naveena B; Brewer, Misty; Duff, Ashley; Lightfoot, Stan; Brush, Richard S; Anderson, Robert E; Rao, Chinthalapally V

    2012-12-01

    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-week-old female p48(Cre/+)-LSL-Kras(G12D/+) and compound Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) 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-p48(Cre/+)-LSL-Kras(G12D/+) mice compared to p48(Cre/+)-LSL-Kras(G12D/+) 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 p48(Cre/+)-LSL-Kras(G12D/+) 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-p48(Cre/+)-LSL-Kras(G12D/+) mice compared to p48(Cre/+)-LSL-Kras(G12D/+) mice. These data highlight the promise of dietary n-3 FAs for chemoprevention of pancreatic cancer in high-risk individuals.

  13. Targeted expression in zebrafish primordial germ cells by Cre/loxP and Gal4/UAS systems.

    PubMed

    Xiong, Feng; Wei, Zhi-Qiang; Zhu, Zuo-Yan; Sun, Yong-Hua

    2013-10-01

    In zebrafish and other vertebrates, primordial germ cells (PGCs) are a population of embryonic cells that give rise to sperm and eggs in adults. Any type of genetically manipulated lines have to be originated from the germ cells of the manipulated founders, thus it is of great importance to establish an effective technology for highly specific PGC-targeted gene manipulation in vertebrates. In the present study, we used the Cre/loxP recombinase system and Gal4/UAS transcription system for induction and regulation of mRFP (monomer red fluorescent protein) gene expression to achieve highly efficient PGC-targeted gene expression in zebrafish. First, we established two transgenic activator lines, Tg(kop:cre) and Tg(kop:KalTA4), to express the Cre recombinases and the Gal4 activator proteins in PGCs. Second, we generated two transgenic effector lines, Tg(kop:loxP-SV40-loxP-mRFP) and Tg(UAS:mRFP), which intrinsically showed transcriptional silence of mRFP. When Tg(kop:cre) females were crossed with Tg(kop:loxP-SV40-loxP-mRFP) males, the loxP flanked SV40 transcriptional stop sequence was 100 % removed from the germ cells of the transgenic hybrids. This led to massive production of PGC-specific mRFP transgenic line, Tg(kop:loxP-mRFP), from an mRFP silent transgenic line, Tg(kop:loxP-SV40-loxP-mRFP). When Tg(kop:KalTA4) females were crossed with Tg(UAS:mRFP) males, the hybrid embryos showed PGC specifically expressed mRFP from shield stage till 25 days post-fertilization (pf), indicating the high sensitivity, high efficiency, and long-lasting effect of the Gal4/UAS system. Real-time PCR analysis showed that the transcriptional amplification efficiency of the Gal4/UAS system in PGCs can be about 300 times higher than in 1-day-pf embryos. More importantly, when the UAS:mRFP-nos1 construct was directly injected into the Tg(kop:KalTA4) embryos, it was possible to specifically label the PGCs with high sensitivity, efficiency, and persistence. Therefore, we have established two

  14. Welfare reform: challenges for grandparents raising grandchildren.

    PubMed

    Copen, Casey E

    2006-01-01

    This paper examines the impact of Temporary Aid to Needy Families (TANF) on a growing constituency that may require welfare assistance: grandparents raising grandchildren. A brief review of the demographics of grandparent caregivers is followed by an exploration of welfare reform legislation, including work requirements and the Healthy Marriage Initiative (HMI). The importance of state-level policies in assisting grandparent-headed households, particularly health care, school enrollment, and housing policies, are also discussed. The paper concludes by offering implications for welfare policies in terms of their impact on intergenerational households headed by grandparents.

  15. 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).

  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, 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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.

  2. BTS strategic plan executive summary

    SciTech Connect

    1999-01-06

    This Strategic Plan positions the Office of Building Technology, State and Community Programs (BTS) to be more effective in reducing energy consumption in homes, offices, schools and other buildings, and in reducing environmental impacts associated with energy use.

  3. A process for implementation of the strategic plan: from strategic planning toward strategic management.

    PubMed

    Chenoy, N C; Morrison, M J; Carlow, D R

    1991-01-01

    Although the principles and process for developing a strategic plan are well established, little information exists on how organizations can implement these plans. The authors developed a framework and methodology for The Ontario Cancer Institute/Princess Margaret Hospital that connects the implementation of the strategic plan with resource allocation and the budget; thus, the budget is a quantified expression of the strategic plan. To accomplish this, a specific cycle of events leading to the development of a corporate calendar was undertaken, which results in the establishment of the annual budget.

  4. Raising children in America: Korean parents' experiences.

    PubMed

    Choi, H; Dancy, B L; Lee, J

    2013-08-01

    This qualitative description study was designed to describe Korean American parents' perceptions of challenges and difficulties they encounter while raising their children in the USA. A convenience sampling of 21 parents of adolescents aged 11-14 years recruited from the Midwest Korean American community participated in the study. Data were collected using in-depth, face-to-face interviews, which took place in agreed-upon, convenient locations. All interviews were audiotaped and transcribed in Korean and the transcripts were translated into English. Qualitative content analysis revealed that the main stresses that parents encountered while raising their children in the USA were inability to advocate for children, feeling uneasy and insecure about incompatible American culture, ambivalence towards children's ethnic identities, and feeling alienated. In relation to these stresses, parents often felt inadequate, ashamed, guilty, regretful and powerless. The findings demonstrated the importance of understanding parents' feelings that are deeply embedded in the conflicted parent-child relationships and their perceptions of being parents in the USA. The present study highlights the need for and importance of providing intervention programmes for parents, particularly programmes that would empower parents, strengthen parent-child relationships and address ways to integrate two very different cultures while upholding ethnic identity and pride.

  5. Executive presence for strategic influence.

    PubMed

    Shirey, Maria R

    2013-01-01

    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 discusses cultivating executive presence, a crucial component of great leadership, needed for strategic influence and to drive change.

  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. 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.

  8. Cre recombinase-mediated restoration of nigrostriatal dopamine in dopamine-deficient mice reverses hypophagia and bradykinesia.

    PubMed

    Hnasko, Thomas S; Perez, Francisco A; Scouras, Alex D; Stoll, Elizabeth A; Gale, Samuel D; Luquet, Serge; Phillips, Paul E M; Kremer, Eric J; Palmiter, Richard D

    2006-06-01

    A line of dopamine-deficient (DD) mice was generated to allow selective restoration of normal dopamine signaling to specific brain regions. These DD floxed stop (DDfs) mice have a nonfunctional Tyrosine hydroxylase (Th) gene because of insertion of a NeoR gene flanked by lox P sites targeted to the first intron of the Th gene. DDfs mice have trace brain dopamine content, severe hypoactivity, and aphagia, and they die without intervention. However, they can be maintained by daily treatment with l-3,4-dihydroxyphenylalanine (L-dopa). Injection of a canine adenovirus (CAV-2) engineered to express Cre recombinase into the central caudate putamen restores normal Th gene expression to the midbrain dopamine neurons that project there because CAV-2 efficiently transduces axon terminals and is retrogradely transported to neuronal cell bodies. Bilateral injection of Cre recombinase into the central caudate putamen restores feeding and normalizes locomotion in DDfs mice. Analysis of feeding behavior by using lickometer cages revealed that virally rescued DDfs mice are hyperphagic and have modified meal structures compared with control mice. The virally rescued DDfs mice are also hyperactive at night, have reduced motor coordination, and are thigmotactic compared with controls. These results highlight the critical role for dopamine signaling in the dorsal striatum for most dopamine-dependent behaviors but suggest that dopamine signaling in other brain regions is important to fine-tune these behaviors. This approach offers numerous advantages compared with previous models aimed at examining dopamine signaling in discrete dopaminergic circuits.

  9. 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.

  10. Impact of tamoxifen on adipocyte lineage tracing: Inducer of adipogenesis and prolonged nuclear translocation of Cre recombinase

    PubMed Central

    Ye, Risheng; Wang, Qiong A.; Tao, Caroline; Vishvanath, Lavanya; Shao, Mengle; McDonald, Jeffery G.; Gupta, Rana K.; Scherer, Philipp E.

    2015-01-01

    Background The selective estrogen receptor modulator tamoxifen, in combination with the Cre-ERT2 fusion protein, has been one of the mainstream methods to induce genetic recombination and has found widespread application in lineage tracing studies. Methods & results Here, we report that tamoxifen exposure at widely used concentrations remains detectable by mass-spectrometric analysis in adipose tissue after a washout period of 10 days. Surprisingly, its ability to maintain nuclear translocation of the Cre-ERT2 protein is preserved beyond 2 months of washout. Tamoxifen treatment acutely leads to transient lipoatrophy, followed by de novo adipogenesis that reconstitutes the original fat mass. In addition, we find a “synthetically lethal” phenotype for adipocytes when tamoxifen treatment is combined with adipocyte-specific loss-of-function mutants, such as an adipocyte-specific PPARγ knockout. This is observed to a lesser extent when alternative inducible approaches are employed. Conclusions These findings highlight the potential for tamoxifen-induced adipogenesis, and the associated drawbacks of the use of tamoxifen in lineage tracing studies, explaining the discrepancy in lineage tracing results from different systems with temporal control of gene targeting. PMID:26629402

  11. 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. PMID:18973000

  12. PCR analysis of chloroplast double-strand break (DSB) repair products induced by I-CreII in Chlamydomonas and Arabidopsis.

    PubMed

    Kwon, Taegun; Odom, Obed W; Qiu, Weihua; Herrin, David L

    2014-01-01

    Homing endonuclease I-CreII has been used to study the consequences and repair of a double-strand break (DSB) in the chloroplast genome of Chlamydomonas and Arabidopsis. Since I-CreII is from a mobile psbA intron of Chlamydomonas, it cleaves the psbA gene of an intronless-psbA strain of Chlamydomonas. And it cleaves specifically in the psbA gene of Arabidopsis, which is naturally intronless. We have shown further that most of the repair products of an I-CreII-induced break in chloroplast DNA can be defined by PCR analysis with total nucleic acids and the appropriate primers. Here, we provide protocols for small-scale preparation of nucleic acids from Chlamydomonas and Arabidopsis, as well as guidelines for the subsequent PCR analysis.

  13. 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

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

    PubMed

    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

  15. 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.

  16. 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)

  17. 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)

  18. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    Not Available

    1992-08-15

    The Strategic Petroleum Reserve (SPR) was created pursuant to the Energy Policy and Conservation Act (EPCA) Public Law 94-163, approved on December 22, 1975, and extended in July 1985, June 1989, March 1990, and September 1990, to reduce the impact of disroptions in petroleum supplies and to carry out obligations of the United States under the Agreement on an International Energy Program. 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. Since the Strategic Petroleum Reserve crude oil storage facilities program for the 750 minion barrels was completed in 1991, this August 15, 1992, Strategic Petroleum Reserve Quarterly Report focuses on activities related primarily to the storage facilities status, oil acquisition, budget, and cost of the Reserve during the period April 1, 1992, through June 30, 1992.

  19. 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. PMID:27387217

  20. 76 FR 14950 - Closed Meeting of the U.S. Strategic Command Strategic Advisory Group

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-18

    ... Commander, U.S. Strategic Command, during the development of the Nation's strategic war plans. Agenda... Destruction, Intelligence Operations, Cyber Operations, Global Strike, Command and Control, Science...

  1. 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

  2. 12. MESSRS. MAUREK AND QUIST RAISING GATE TO ALLOW FIRST ...

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

    12. MESSRS. MAUREK AND QUIST RAISING GATE TO ALLOW FIRST WATER TO PASS (IT TOOK SIX MEN 30 MINUTES TO RAISE THE GATE FIVE FEET) - Upper Souris National Wildlife Refuge, Dam 96, Souris River Basin, Foxholm, Surrey (England), ND

  3. 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.

  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 Asia 2002 Final Report

    SciTech Connect

    Richard Ellings; Aaron Friedberg; Michael Wills

    2002-09-01

    The Strategic Asia Program made considerable progress over the course of 2002--the program's first year with support from the Department of Energy--and completed all its tasks on schedule and within budget. Following a planning meeting in Washington in February 2002, a team of leading specialists wrote a series of original assessments regarding the impact of September 11 on the strategic environment in Asia, examining how perceptions and strategies of countries in the region changed following the terrorist attacks. The final products, Strategic Asia 2002-03: Asian Aftershocks and its accompanying executive summary, were published in September 2002. The program's research findings (some of which are summarized) were presented to policymakers in Washington and elsewhere throughout the year, and almost 2,000 copies of the book had been distributed by mid-2003.

  6. Raising the visibility of school nursing services.

    PubMed

    Sanders, Laura; Pratten, Charlie; Pretty, Laura; Roberts, Sheila; Robinson, Jenny

    2016-06-01

    In an attempt to raise the visibility and profile of the school nursing service in the London boroughs of Hounslow and Richmond, school nurses sought to hear from the voices of the young people with whom they work. Ten focus groups in secondary educational provisions were held in a bid to look at young people's accessibility to technological devices, where they would go if they had a health problem and which health topics they would like more information on. This paper examines both national and local data and demographics, and discusses findings from the focus groups run with local young people. The results collected from the focus groups emphasised that technology is a fundamental aspect of many young people's lives and that a large number of young people access the internet to source information about their personal health and wellbeing. The data suggests that the school nursing service could use technologies, such as an interactive app, to support the physical and emotional health of young people. PMID:27443030

  7. 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.

  8. 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.

  9. Development of pronuclear injection-based targeted transgenesis in mice through Cre-loxP site-specific recombination.

    PubMed

    Ohtsuka, Masato

    2014-01-01

    Microinjection of DNA into the pronuclei of zygotes is the simplest and most widely used method for generating transgenic (Tg) mice. However, it is always associated with random integration of multiple copies of the transgene, resulting in unstable, low, or no transgene expression due to positional effects and/or repeat-induced gene silencing. In addition, random integration sometimes disrupts an endogenous gene that can affect the phenotypes of Tg mice. Our recently developed pronuclear injection-based targeted transgenesis (PITT) method enables the integration of a single-copy transgene into a predetermined genomic locus through Cre-loxP site-specific recombination. The PITT method enables stable and reliable transgene expression in Tg mice and is also applicable for generating knockdown mice. Therefore, the PITT method could represent next-generation transgenesis that overcomes the pitfalls of conventional transgenesis.

  10. 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)

  11. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 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...

  12. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 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...

  13. 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...

  14. 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...

  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, 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 5 CFR 2421.21 - Affected by issues raised.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Affected by issues raised. 2421.21 Section 2421.21 Administrative Personnel FEDERAL LABOR RELATIONS AUTHORITY, GENERAL COUNSEL OF THE FEDERAL... § 2421.21 Affected by issues raised. The phrase affected by issues raised, as used in part 2422,...

  5. 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…

  6. 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…

  7. 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.

  8. 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

  9. 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.

  10. 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; 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: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

  11. 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.

  12. The CreA repressor is the sole DNA-binding protein responsible for carbon catabolite repression of the alcA gene in Aspergillus nidulans via its binding to a couple of specific sites.

    PubMed

    Panozzo, C; Cornillot, E; Felenbok, B

    1998-03-13

    Carbon catabolite repression is mediated in Aspergillus nidulans by the negative acting protein CreA. The CreA repressor plays a major role in the control of the expression of the alc regulon, encoding proteins required for the ethanol utilization pathway. It represses directly, at the transcriptional level, the specific transacting gene alcR, the two structural genes alcA and aldA, and other alc genes in all physiological growth conditions. Among the seven putative CreA sites identified in the alcA promoter region, we have determined the CreA functional targets in AlcR constitutive and derepressed genetic backgrounds. Two different divergent CreA sites, of which one overlaps a functional AlcR inverted repeat site, are largely responsible for alcA repression. Totally derepressed alcA expression is achieved when these two CreA sites are disrupted in addition to another single site, which overlaps the functional palindromic induction target. The fact that derepression is always associated with alcA overexpression is consistent with a competition model between AlcR and CreA for their cognate targets in the same region of the alcA promoter. Our results also indicate that the CreA repressor is necessary and sufficient for the total repression of the alcA gene.

  13. Divide and conquer: strategic decision areas

    PubMed Central

    Kolling, Nils; Hunt, Laurence T

    2016-01-01

    Human decisions are often strategic, but this can prove difficult to study experimentally. The board game shogi is used to investigate the functional neuroanatomy of strategic decisions, revealing different brain areas to other forms of choice. PMID:25919959

  14. A Primer on Strategic Financial Assessments.

    ERIC Educational Resources Information Center

    Richman, Naomi; Fitzgerald, Susan

    2003-01-01

    Describes how to perform a strategic financial assessment to enable the board to understand the fundamental internal and external challenges and opportunities confronting the institution when decision making and strategic capital planning. (EV)

  15. Removal of selectable marker gene from fibroblast cells in transgenic cloned cattle by transient expression of Cre recombinase and subsequent effects on recloned embryo development.

    PubMed

    Wang, S; Sun, X; Ding, F; Zhang, K; Zhao, R; Li, S; Li, R; Tang, B; Zhang, L; Liu, Y; Li, J; Gao, F; Wang, H; Wang, L; Dai, Y; Li, N

    2009-09-01

    Introduction of selectable marker genes to transgenic animals could create an inconvenience to further research and may exaggerate public concerns regarding biological safety. The objective of the current study was to excise loxP flanked neo(R) in transgenic cloned cattle by transient expression of Cre recombinase. Green fluorescent protein gene (GFP) was incorporated to monitor Cre expression; therefore, Cre-expressed cells could be selected indirectly by fluorescence-activated cell sorting (FACS). The neo(R) was removed and Cre expressed transiently in GFP-positive colonies; excision of neo(R) was confirmed by single-blastocyst PCR in recloned blastocysts, with neo(R)-free fibroblast cells as donors. There was no difference (P>0.05) in rates of cleavage (76.0% vs. 68.8%) or blastocyst formation (56.6% vs. 52.9%) between recloned embryos with neo(R)-free or neo(R)-included donors. The differential staining of recloned blastocysts were similar (P >0.05) in terms of total cell number (124 vs. 122) and the ratio of ICM (Inner Cell Mass) to the total cell number (38.1% vs. 38.2%). Furthermore, pregnancy and calving rates were not different (P>0.05) from those of the control. In conclusion, we successfully excised neo(R) from transgenic cloned cattle; the manipulation did not affect the developmental competence of recloned preimplantation embryos. This approach should benefit bioreactor and transgenic research in livestock.

  16. 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

  17. 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. PMID:21876744

  18. 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.

  19. 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

  20. Strategic Listening for School Leaders

    ERIC Educational Resources Information Center

    Tate, Jeannine S.; Dunklee, Dennis R.

    2005-01-01

    The ability to communicate effectively with multiple constituencies is recognized as an essential characteristic of effective leaders. Listening strategically is a way of showing parents, students, faculty, staff, and others that their ideas and beliefs are of value. The authors' practitioner-friendly book concentrates on the importance of…

  1. 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.

  2. Strategic Planning of Technology Transfer.

    ERIC Educational Resources Information Center

    Groff, Warren H.

    Using the Ohio Technology Transfer Organization (OTTO) as its primary example, this paper offers a strategic planning perspective on technology transfer and human resources development. First, a brief overview is provided of the maturation of mission priorities and planning processes in higher education in the United States, followed by a…

  3. Strategic Approaches with Resistant Families.

    ERIC Educational Resources Information Center

    Breit, Miranda; And Others

    1983-01-01

    Describes the operation of a 10-session brief therapy unit for families who have failed in more traditional treatment modalities. Case material is presented to exemplify five different treatment strategies: symptom prescription, reframing, illusion of alternatives, role play, and strategic alliances. Advantages and limitations are discussed.…

  4. 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…

  5. 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.

  6. 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)

  7. Strategic Planning: The Catastrophic Approach.

    ERIC Educational Resources Information Center

    Brody, Stanley J.

    1987-01-01

    Reviews and defines continuity of care, or long term care, as a potential economic catastrophe which requires strategic planning. Proposes a Medicare revision that provides integrated funding for acute, sub-acute, and long-term care as elements of continuity of care. (Author/ABB)

  8. 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…

  9. Strategic Plan 2013 to 2016

    ERIC Educational Resources Information Center

    Lumina Foundation for Education, 2013

    2013-01-01

    In 2009, Lumina Foundation released its first strategic plan, based on the goal that 60% of Americans obtain a high-quality postsecondary degree or credential by 2025-- a goal now known as Goal 2025. Much has changed even in the short time since that plan was written, both in the external environment and in what has been learned from this work.…

  10. Strategic Planning for School Success.

    ERIC Educational Resources Information Center

    Herman, Jerry J.

    1993-01-01

    Strategic planners concerned with such matters as high-achieving students, high-performing teachers, broad-based community support, and a two-way involvement with the community must analyze the strengths, weaknesses, opportunities, and threats existing in the school's internal and external environment. A sample SWOT analysis is included. (MLH)

  11. 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:…

  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 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…

  14. Strategic Change in AAS Publishing

    NASA Astrophysics Data System (ADS)

    Steffen, Julie

    2015-08-01

    The American Astronomical Society has embarked on a process of strategic change in its publishing program. The process has incuded authors, AAS leaders, editors, publishing experts, librarians, and data scientists. This session will outline the still ongoing process and present some both upcoming and already available new AAS Publishing features and services to the global astronomy community.

  15. 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…

  16. 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.

  17. 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

  18. 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

  19. 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

  20. 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…

  1. 77 FR 54615 - Strategic Management Program; Fiscal Year 2013-2016 Strategic Plan

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-05

    ... SAFETY BOARD Strategic Management Program; Fiscal Year 2013-2016 Strategic Plan AGENCY: National... on the proposed strategic plan to be published by October 2012. All interested parties are invited to submit comments regarding this proposed strategic plan. As background, the NTSB's 2010-2015...

  2. 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…

  3. 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…

  4. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 23 Highways 1 2011-04-01 2011-04-01 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...

  5. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 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...

  6. 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...

  7. 12 CFR 195.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 1 2013-01-01 2013-01-01 false Strategic plan. 195.27 Section 195.27 Banks and... Assessing Performance § 195.27 Strategic plan. (a) Alternative election. The appropriate Federal banking... area(s) under a strategic plan if: (1) The savings association has submitted the plan to...

  8. 12 CFR 345.27 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 4 2011-01-01 2011-01-01 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)...

  9. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 3 2012-01-01 2012-01-01 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...

  10. 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...

  11. 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...

  12. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 23 Highways 1 2010-04-01 2010-04-01 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...

  13. 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...

  14. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 6 2013-01-01 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...

  15. 12 CFR 25.27 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 1 2011-01-01 2011-01-01 false Strategic plan. 25.27 Section 25.27 Banks and... DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the OCC as provided...

  16. 12 CFR 25.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 1 2014-01-01 2014-01-01 false Strategic plan. 25.27 Section 25.27 Banks and... DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the OCC as provided...

  17. 12 CFR 345.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 5 2013-01-01 2013-01-01 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)...

  18. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 3 2011-01-01 2011-01-01 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...

  19. 12 CFR 345.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 5 2014-01-01 2014-01-01 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)...

  20. 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...

  1. 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...

  2. 12 CFR 195.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 1 2012-01-01 2012-01-01 false Strategic plan. 195.27 Section 195.27 Banks and... Assessing Performance § 195.27 Strategic plan. (a) Alternative election. The appropriate Federal banking... area(s) under a strategic plan if: (1) The savings association has submitted the plan to...

  3. 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...

  4. 12 CFR 195.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 1 2014-01-01 2014-01-01 false Strategic plan. 195.27 Section 195.27 Banks and... Assessing Performance § 195.27 Strategic plan. (a) Alternative election. The appropriate Federal banking... area(s) under a strategic plan if: (1) The savings association has submitted the plan to...

  5. 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...

  6. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 5 2011-01-01 2011-01-01 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...

  7. 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...

  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. 12 CFR 25.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 1 2013-01-01 2013-01-01 false Strategic plan. 25.27 Section 25.27 Banks and... DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the OCC as provided...

  10. 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...

  11. 12 CFR 25.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 1 2012-01-01 2012-01-01 false Strategic plan. 25.27 Section 25.27 Banks and... DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the OCC as provided...

  12. 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)...

  13. 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.…

  14. 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…

  15. A Cre-lox approach for transient transgene expression in neural precursor cells and long-term tracking of their progeny in vitro and in vivo

    PubMed Central

    Geoffroy, Cédric G; Raineteau, Olivier

    2007-01-01

    Background Neural precursor cells (NPCs) can be isolated from various regions of the postnatal central nervous system (CNS). Manipulation of gene expression in these cells offers a promising strategy to manipulate their fate both in vitro and in vivo. In this study, we developed a technique that allows the transient manipulation of single/multiple gene expression in NPCs in vitro, and the long-term tracking of their progeny both in vitro and in vivo. Results In order to combine the advantages of transient transfection with the long-term tracking of the transfected cells progeny, we developed a new approach based on the cre-lox technology. We first established a fast and reliable protocol to isolate and culture NPCs as monolayer, from the spinal cord of neonatal transgenic Rosa26-YFP cre-reporter mice. These cells could be reliably transfected with single/multiple plasmids by nucleofection. Nucleofection with mono- or bicistronic plasmids containing the Cre recombinase gene resulted in efficient recombination and the long-term expression of the YFP-reporter gene. The transient cre-expression was non-toxic for the transfected cells and did not alter their intrinsic properties. Finally, we demonstrated that cre-transfected cells could be transplanted into the adult brain, where they maintained YFP expression permitting long-term tracking of their migration and differentiation. Conclusion This approach allows single/multiple genes to be manipulated in NPCs, while at the same time allowing long-term tracking of the transfected cells progeny to be analyzed both in vitro and in vivo. PMID:17504531

  16. Nkx2.2:Cre knock-in mouse line: a novel tool for pancreas- and CNS-specific gene deletion.

    PubMed

    Balderes, Dina A; Magnuson, Mark A; Sussel, Lori

    2013-12-01

    Nkx2.2 is a homeodomain-containing transcriptional regulator necessary for the appropriate differentiation of ventral neuronal populations in the spinal cord and hindbrain, and endocrine cell populations in the pancreas and intestine. In each tissue, Nkx2.2 inactivation leads to reciprocal cell fate alterations. To confirm the cell fate changes are due to respecification of Nkx2.2-expressing progenitors and to provide a novel tool for lineage tracing in the pancreas and CNS, we generated an Nkx2.2:Cre mouse line by knocking in a Cre-EGFP cassette into the Nkx2.2 genomic locus and inactivating endogenous Nkx2.2. The R26R-CAG-LSL-tdTomato reporter was used to monitor the specificity and efficiency of Nkx2.2:Cre activity; the tomato reporter faithfully recapitulated endogenous Nkx2.2 expression and could be detected as early as embryonic day (e) 9.25 in the developing CNS and was initiated shortly thereafter at e9.5 in the pancreas. Lineage analyses in the CNS confirmed the cell populations thought to be derived from Nkx2.2-expressing progenitor domains. Furthermore, lineage studies verified Nkx2.2 expression in the earliest pancreatic progenitors that give rise to all cell types of the pancreas; however they also revealed more robust Cre activity in the dorsal versus ventral pancreas. Thus, the Nkx2.2:Cre line provides a novel tool for gene manipulations in the CNS and pancreas.

  17. The CReSIS Radar Suite for Measurements of the Ice Sheets and Sea Ice during Operation Ice Bridge

    NASA Astrophysics Data System (ADS)

    Leuschen, C.; Gogineni, P. S.; Allen, C.; Paden, J. D.; Hale, R.; Rodriguez-Morales, F.; Harish, A.; Seguin, S.; Arnold, E.; Blake, W.; Byers, K.; Crowe, R.; Lewis, C.; Panzer, B.; Patel, A.; Shi, L.

    2010-12-01

    The University of Kansas, Center for Remote Sensing of Ice Sheets (CReSIS) has developed a suite of radar instrumentation operating at frequencies ranging from 180 MHz to 18 GHz to monitor the major Greenland and Antarctic ice sheets at varying resolution from the surface to the bed as well as the surface topography and snow cover characteristics of sea ice. The radar suite includes a VHF multi-channel coherent radar depth sounder/imager (MCoRDS/I) to measure ice sheet thickness, internal layering, and image ice-bed interface with fine resolution; a UHF accumulation radar to measure shallow (up to 500 m) ice sheet thickness, internal layering, and accumulation rates at decimeter resolutions; an ultra-wideband “snow” radar to measure thickness of snow over sea ice and accumulation rates at centimeter scale resolution; and a wideband Ku-band altimeter to measure surface topography and accumulation layer for comparison with CryoSat-2. To support recent NASA Operation Ice Bridge activities, CReSIS radars were adapted for installation and operation on the NASA DC-8 for deployments primarily to Antarctica for measurements over glacial ice in West Antarctica and sea ice, and on the NASA P-3 for deployments to Greenland and Arctic sea ice. The major integration activities included the development of antenna subsystems specifically tailored for the DC-8 and P-3 aircraft. This included a low-profile radar depth sounder/imager antenna assembly for the DC-8 with a minimal impact on the aircraft range, one of the largest VHF antenna arrays for the P-3, and a set of internal antennas structures and radomes for the P-3 bomb bay. The design and development of the antenna assemblies were focused on meeting a combination of instrument and mission requirements. We will present a description of each radar system including the antenna assemblies, instrument block diagrams, modes of operation, installation specifics for each OIB deployment, example results, and availability of data

  18. 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.

  19. 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.

  20. Strategic alliances and market risk.

    PubMed

    Havenaar, Matthias; Hiscocks, Peter

    2012-08-01

    Strategic alliances in product development and marketing are crucial to the biotechnology industry. Many alliances, however, are terminated before the drug reaches the market. In this article we make the case that strategic alliances can fail because of how they are negotiated. Alliance contracts are often inflexible and do not allow for changes in market conditions. We propose a model for contract valuation that can assist biotech and/or pharma deal makers in negotiating alliances that have a higher chance of survival in uncertain market conditions. The model makes use of variable royalties and milestone payments. Because licensing is key to the biotech and/or pharma business model this article will be of interest not only to professionals in licensing, but to all professionals active in the industry. PMID:22484547

  1. Rehabilitation: strategic, proactive, flexible, risky?

    PubMed

    Green, R H

    2000-12-01

    Rehabilitation after armed conflict is a direct intellectual descendant of thinking about rehabilitation after natural calamity. It is related, generally, to poverty reduction and, operationally, to associated action at the micro level. This history has limited its strategic conceptualisation and, in particular, its links with reconciliation and state re-legitimation and also with macro-economic stabilisation and renewed growth. In post-war--or more generally, a lull in conflict with the potential to become permanent--a country's rapid, focused, prioritised action within a strategic framework is urgent. It is not risk-free for political and natural disasters as well as for those with economic and social bases. Flexibility, learning from initial experience and asking intended household beneficiaries about their needs in advance can reduce risk as can pre-positioning of contingency or standby resources to avert post-war calamities (for example, drought, flood) and catastrophes (renewed violence) from delaying and discrediting rehabilitation efforts.

  2. 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.

  3. 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. PMID:27318284

  4. The strategic importance of data.

    PubMed

    Glaser, John; Markell, Peter; Stone, John

    2010-02-01

    Two factors--accountability for care and an increased adoption of interoperable electronic health records (EHRs)--will significantly increase the importance of data for healthcare organizations. These factors have significant data ramifications in four areas: payment reform, data use policies, IT strategy, and data management. As data become a strategic asset, healthcare finance executives will need to become significant contributors to their organizations' EHR initiatives and data management strategies. PMID:20178239

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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

    ... overall CMV transportation life-cycle. The CMV transportation life-cycle concept encompasses the whole CMV... are part of the CMV transportation life-cycle need to be aware of their impact on CMV safety and take... transportation life-cycle. Recognize that, while safety is FMCSA's highest priority, the Agency must also...

  10. Pten deletion in RIP-Cre neurons protects against type 2 diabetes by activating the anti-inflammatory reflex.

    PubMed

    Wang, Linyuan; Opland, Darren; Tsai, Sue; Luk, Cynthia T; Schroer, Stephanie A; Allison, Margaret B; Elia, Andrew J; Furlonger, Caren; Suzuki, Akira; Paige, Christopher J; Mak, Tak W; Winer, Daniel A; Myers, Martin G; Woo, Minna

    2014-05-01

    Inflammation has a critical role in the development of insulin resistance. Recent evidence points to a contribution by the central nervous system in the modulation of peripheral inflammation through the anti-inflammatory reflex. However, the importance of this phenomenon remains elusive in type 2 diabetes pathogenesis. Here we show that rat insulin-2 promoter (Rip)-mediated deletion of Pten, a gene encoding a negative regulator of PI3K signaling, led to activation of the cholinergic anti-inflammatory pathway that is mediated by M2 activated macrophages in peripheral tissues. As such, Rip-cre(+) Pten(flox/flox) mice showed lower systemic inflammation and greater insulin sensitivity under basal conditions compared to littermate controls, which were abolished when the mice were treated with an acetylcholine receptor antagonist or when macrophages were depleted. After feeding with a high-fat diet, the Pten-deleted mice remained markedly insulin sensitive, which correlated with massive subcutaneous fat expansion. They also exhibited more adipogenesis with M2 macrophage infiltration, both of which were abolished after disruption of the anti-inflammatory efferent pathway by left vagotomy. In summary, we show that Pten expression in Rip(+) neurons has a critical role in diabetes pathogenesis through mediating the anti-inflammatory reflex.

  11. 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

  12. The Small Library and Fund-Raising for Automation.

    ERIC Educational Resources Information Center

    Burlingame, Dwight F.

    1989-01-01

    Offers advice on the principles of successful fund raising, specifically addressing small libraries and fund raising for automation. The discussion covers the need for careful planning, identification of receptive funding sources, and clear communication of needs and objectives. A sidebar provides information on the Foundation Center, an…

  13. 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…

  14. An Introduction to Fund Raising: The Newcomers' Guide to Development.

    ERIC Educational Resources Information Center

    Faust, Paula J., Ed.

    A comprehensive introduction to all phases of college fund-raising is presented in 14 chapters. Attention is directed to the philosophy behind development, development's role in institutional advancement, and the personal qualities that lead to success in fund-raising. Basic principles also are covered for the following: annual giving, the capital…

  15. 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…

  16. 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

  17. 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…

  18. 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

  19. The health of grandparents raising grandchildren: a literature review.

    PubMed

    Hadfield, Jessica C

    2014-04-01

    Around the world, an increasing number of grandparents are raising grandchildren in households without a biological parent. This is no less true in the United States, where grandparents raising grandchildren are at increased risk for depression and declining physical health. The purpose of this article is to explore the recent literature as it relates to the psychological and/or physical health of grandparents raising grandchildren. On reviewing 19 articles from the past 10 years, it is clear that the literature consistently verifies the health risks, especially depression, for grandparents raising grandchildren; however, the lack of research regarding grandfathers and non-African American caregivers is apparent. The factors influencing grandparents raising grandchildren are numerous, which calls for care providers to assess the needs, situations, and perceptions of grandparents individually. Finally, a need has arisen for more research suggesting and validating interventions that health care providers can use to support this population of older adults. PMID:24568146

  20. A Strategic Approach to Aggression.

    ERIC Educational Resources Information Center

    Archer, John

    2001-01-01

    Discusses two issues raised by Underwood et al.: the distinction between indirect and relational forms of aggression, and implications of indirect aggression for definitions of aggression; and the normative view of aggression that indicates that aggressive individuals may be socially skilled. Suggests that both issues lead to the conclusion that…

  1. 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)

  2. cAMP target sequences enhCRE and CNRE sense low-salt intake to increase human renin gene expression in vivo.

    PubMed

    Desch, Michael; Harlander, Sabine; Neubauer, Björn; Gerl, Melanie; Germain, Stephane; Castrop, Hayo; Todorov, Vladimir T

    2011-05-01

    This study aimed to assess the role of cAMP target sequences enhancer cAMP response element (enhCRE) and cAMP and overlapping negative response element (CNRE) in the control of human renin gene (REN) in vivo. enhCRE and CNRE were silenced by mutations in a 12.2-kb human renin promoter fused to LacZ reporter gene. This construct was used to generate transgenic mice (RENMut-LacZ). The expression of the transgene was correctly targeted to the juxtaglomerular portions of renal afferent arterioles which express endogenous mouse renin. Therefore, enhCRE and CNRE do not seem to be relevant for the control of the cell-specific expression of the human renin gene. The β-adrenoreceptor agonist isoproterenol (10 mg/kg/day, for 2 days) stimulated the endogenous renin, but not the LacZ mRNA expression. Treatment of RENMut-LacZ mice with the angiotensin converting enzyme inhibitor (enalapril 10 mg/kg/day, for 7 days) or their crossing to angiotensin receptor type 1a knockout mice led to increased renin and LacZ mRNA levels. Renin expression was upregulated by low-salt diet (0.03% NaCl, for 10 days) and downregulated by high-salt diet (4% NaCl, for 10 days). In contrast, low-salt diet did not influence, while high-salt diet inhibited the expression of LacZ. In summary, enhCRE and CNRE appear to be necessary for the transactivation of the human renin gene through β-adrenoreceptors and by low-salt diet. Our data also suggest that different intracellular mechanisms mediate the effect of low- and high-salt intake on renin expression in vivo.

  3. Metabolic flux analysis of Escherichia coli creB and arcA mutants reveals shared control of carbon catabolism under microaerobic growth conditions.

    PubMed

    Nikel, Pablo I; Zhu, Jiangfeng; San, Ka-Yiu; Méndez, Beatriz S; Bennett, George N

    2009-09-01

    Escherichia coli has several elaborate sensing mechanisms for response to availability of oxygen and other electron acceptors, as well as the carbon source in the surrounding environment. Among them, the CreBC and ArcAB two-component signal transduction systems are responsible for regulation of carbon source utilization and redox control in response to oxygen availability, respectively. We assessed the role of CreBC and ArcAB in regulating the central carbon metabolism of E. coli under microaerobic conditions by means of (13)C-labeling experiments in chemostat cultures of a wild-type strain, DeltacreB and DeltaarcA single mutants, and a DeltacreB DeltaarcA double mutant. Continuous cultures were conducted at D = 0.1 h(-1) under carbon-limited conditions with restricted oxygen supply. Although all experimental strains metabolized glucose mainly through the Embden-Meyerhof-Parnas pathway, mutant strains had significantly lower fluxes in both the oxidative and the nonoxidative pentose phosphate pathways. Significant differences were also found at the pyruvate branching point. Both pyruvate-formate lyase and the pyruvate dehydrogenase complex contributed to acetyl-coenzyme A synthesis from pyruvate, and their activity seemed to be modulated by both ArcAB and CreBC. Strains carrying the creB deletion showed a higher biomass yield on glucose compared to the wild-type strain and its DeltaarcA derivative, which also correlated with higher fluxes from building blocks to biomass. Glyoxylate shunt and lactate dehydrogenase were active mainly in the DeltaarcA strain. Finally, it was observed that the tricarboxylic acid cycle reactions operated in a rather cyclic fashion under our experimental conditions, with reduced activity in the mutant strains.

  4. 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

  5. Intracellular Cre-Mediated Deletion of the Unique Packaging Signal Carried by a Herpes Simplex Virus Type 1 Recombinant and Its Relationship to the Cleavage-Packaging Process

    PubMed Central

    Logvinoff, Carine; Epstein, Alberto L.

    2000-01-01

    To gain further insight on the function of the herpes simplex virus type 1 (HSV-1) packaging signal (a sequence), we constructed a recombinant virus containing a unique a sequence, which was flanked by two loxP sites in parallel orientation. The phenotype of this recombinant, named HSV-1 LaL, was studied in cell lines which either express or do not express Cre recombinase. Although LaL virus multiplication was only slightly reduced in standard cell lines, its growth was strongly inhibited in Cre-expressing cells. In these cells, a sequences were detected mostly in low-molecular-weight DNA circles, indicating that they had been excised from virus DNA by site-specific recombination. Deletion of the a sequences from the viral genome resulted in the accumulation of uncleaved replication intermediates, as observed by pulsed-field gel electrophoresis. B-type capsids also accumulated in these cells, as shown both by electron microscopy and by sucrose gradient sedimentation. Further examination of the status of a sequences in Cre-expressing cells indicated that high-level amplification of this sequence can occur in the absence of the cleavage-packaging process. Moreover, the amplified a signals in small circular DNA molecules remained uncleaved, indicating that these molecules were not able to efficiently interact with the cleavage-packaging machinery. The cleavage-packaging machinery and the structural proteins required to assemble virions were, however, functional in HSV-1 LaL-infected Cre-expressing cells, since this system could be used to package plasmid DNA harboring an origin of virus replication and one normal a signal. This is the first study in which accumulation both of uncleaved replication intermediates and of B capsids has been obtained in the presence of the full set of proteins required to package virus DNA. PMID:10954540

  6. Strategic planning decision making using fuzzy SWOT-TOPSIS with reliability factor

    NASA Astrophysics Data System (ADS)

    Mohamad, Daud; Afandi, Nur Syamimi; Kamis, Nor Hanimah

    2015-10-01

    Strategic planning is a process of decision making and action for long-term activities in an organization. The Strengths, Weaknesses, Opportunities and Threats (SWOT) analysis has been commonly used to help organizations in strategizing their future direction by analyzing internal and external environment. However, SWOT analysis has some limitations as it is unable to prioritize appropriately the multiple alternative strategic decisions. Some efforts have been made to solve this problem by incorporating Multi Criteria Decision Making (MCDM) methods. Nevertheless, another important aspect has raised concerns on obtaining the decision that is the reliability of the information. Decision makers evaluate differently depending on their level of confidence or sureness in the evaluation. This study proposes a decision making procedure for strategic planning using SWOT-TOPSIS method by incorporating the reliability factor of the evaluation based on Z-number. An example using a local authority in the east coast of Malaysia is illustrated to determine the strategic options ranking and to prioritize factors in each SWOT category.

  7. 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.

  8. Psychiatric disorder-related abnormal behavior and habenulointerpeduncular pathway defects in Wnt1-cre and Wnt1-GAL4 double transgenic mice.

    PubMed

    Nakajima, Mitsunari; Mori, Hisamichi; Nishikawa, Chisa; Tsuruta, Momoko; Okuyama, Satoshi; Furukawa, Yoshiko

    2013-01-01

    The neural crest is a unique structure in vertebrates. Wnt1-cre and Wnt1-GAL4 double transgenic (dTg) mice have been used in a variety of studies concerning neural crest cell lineages in which the Cre/loxP or GAL4/UAS system was applied. Here, we show psychiatric disorder-related behavioral abnormalities and histologic alterations in a neural crest-derived brain region in dTg mice. The dTg mice exhibited increased locomotor activity, decreased social interaction, and impaired short-term spatial memory and nesting behavior. The choline acetyltransferase- and vesicular glutamate transporter 2-immunoreactive habenulointerpeduncular fiber tracts that project from the medial habenular nucleus of the epithalamus to the interpeduncular nucleus of the midbrain tegmentum appeared irregular in the dTg mice. Both the medial habenula nucleus and the interpeduncular nucleus were confirmed to be derived from the neural crest. The findings of this study suggest that neural crest-derived cells have pathogenic roles in the development of psychiatric disorders and that the dTg mouse could be a useful animal model for studying the pathophysiology of mental illness such as autism and schizophrenia. Scientists that use the dTg mice as a cre-transgenic deleter line should be cautious in its possible toxicity, especially if behavioral analyses are to be performed.

  9. Mechanosensory and ATP Release Deficits following Keratin14-Cre-Mediated TRPA1 Deletion Despite Absence of TRPA1 in Murine Keratinocytes

    PubMed Central

    Palygin, Oleg; Weyer, Andy D.; Barabas, Marie E.; Lawlor, Michael W.; Staruschenko, Alexander; Stucky, Cheryl L.

    2016-01-01

    Keratinocytes are the first cells that come into direct contact with external tactile stimuli; however, their role in touch transduction in vivo is not clear. The ion channel Transient Receptor Potential Ankyrin 1 (TRPA1) is essential for some mechanically-gated currents in sensory neurons, amplifies mechanical responses after inflammation, and has been reported to be expressed in human and mouse skin. Other reports have not detected Trpa1 mRNA transcripts in human or mouse epidermis. Therefore, we set out to determine whether selective deletion of Trpa1 from keratinocytes would impact mechanosensation. We generated K14Cre-Trpa1fl/fl mice lacking TRPA1 in K14-expressing cells, including keratinocytes. Surprisingly, Trpa1 transcripts were very poorly detected in epidermis of these mice or in controls, and detection was minimal enough to preclude observation of Trpa1 mRNA knockdown in the K14Cre-Trpa1fl/fl mice. Unexpectedly, these K14Cre-Trpa1fl/fl mice nonetheless exhibited a pronounced deficit in mechanosensitivity at the behavioral and primary afferent levels, and decreased mechanically-evoked ATP release from skin. Overall, while these data suggest that the intended targeted deletion of Trpa1 from keratin 14-expressing cells of the epidermis induces functional deficits in mechanotransduction and ATP release, these deficits are in fact likely due to factors other than reduction of Trpa1 expression in adult mouse keratinocytes because they express very little, if any, Trpa1. PMID:26978657

  10. Mesoporous Silica Nanoparticle-Mediated Intracellular Cre Protein Delivery for Maize Genome Editing via loxP Site Excision1,2[W][OPEN

    PubMed Central

    Martin-Ortigosa, Susana; Peterson, David J.; Valenstein, Justin S.; Lin, Victor S.-Y.; Trewyn, Brian G.; Lyznik, L. Alexander; Wang, Kan

    2014-01-01

    The delivery of proteins instead of DNA into plant cells allows for a transient presence of the protein or enzyme that can be useful for biochemical analysis or genome modifications. This may be of particular interest for genome editing, because it can avoid DNA (transgene) integration into the genome and generate precisely modified “nontransgenic” plants. In this work, we explore direct protein delivery to plant cells using mesoporous silica nanoparticles (MSNs) as carriers to deliver Cre recombinase protein into maize (Zea mays) cells. Cre protein was loaded inside the pores of gold-plated MSNs, and these particles were delivered by the biolistic method to plant cells harboring loxP sites flanking a selection gene and a reporter gene. Cre protein was released inside the cell, leading to recombination of the loxP sites and elimination of both genes. Visual selection was used to select recombination events from which fertile plants were regenerated. Up to 20% of bombarded embryos produced calli with the recombined loxP sites under our experimental conditions. This direct and reproducible technology offers an alternative for DNA-free genome-editing technologies in which MSNs can be tailored to accommodate the desired enzyme and to reach the desired tissue through the biolistic method. PMID:24376280

  11. Mechanosensory and ATP Release Deficits following Keratin14-Cre-Mediated TRPA1 Deletion Despite Absence of TRPA1 in Murine Keratinocytes.

    PubMed

    Zappia, Katherine J; Garrison, Sheldon R; Palygin, Oleg; Weyer, Andy D; Barabas, Marie E; Lawlor, Michael W; Staruschenko, Alexander; Stucky, Cheryl L

    2016-01-01

    Keratinocytes are the first cells that come into direct contact with external tactile stimuli; however, their role in touch transduction in vivo is not clear. The ion channel Transient Receptor Potential Ankyrin 1 (TRPA1) is essential for some mechanically-gated currents in sensory neurons, amplifies mechanical responses after inflammation, and has been reported to be expressed in human and mouse skin. Other reports have not detected Trpa1 mRNA transcripts in human or mouse epidermis. Therefore, we set out to determine whether selective deletion of Trpa1 from keratinocytes would impact mechanosensation. We generated K14Cre-Trpa1fl/fl mice lacking TRPA1 in K14-expressing cells, including keratinocytes. Surprisingly, Trpa1 transcripts were very poorly detected in epidermis of these mice or in controls, and detection was minimal enough to preclude observation of Trpa1 mRNA knockdown in the K14Cre-Trpa1fl/fl mice. Unexpectedly, these K14Cre-Trpa1fl/fl mice nonetheless exhibited a pronounced deficit in mechanosensitivity at the behavioral and primary afferent levels, and decreased mechanically-evoked ATP release from skin. Overall, while these data suggest that the intended targeted deletion of Trpa1 from keratin 14-expressing cells of the epidermis induces functional deficits in mechanotransduction and ATP release, these deficits are in fact likely due to factors other than reduction of Trpa1 expression in adult mouse keratinocytes because they express very little, if any, Trpa1. PMID:26978657

  12. Excellence in Educational Fund Raising at America's Community Colleges and a Key Resources Guide for Educational Fund Raising.

    ERIC Educational Resources Information Center

    Ryan, G. Jeremiah

    Designed to encourage and inform community college efforts to secure private financial support, this literature review and resource guide examine the current status of fund raising at community colleges and list pertinent information sources. After introductory comments advocate increased community college involvement in fund raising, the paper…

  13. Strategic workshops on cancer nanotechnology.

    PubMed

    Nagahara, Larry A; Lee, Jerry S H; Molnar, Linda K; Panaro, Nicholas J; Farrell, Dorothy; Ptak, Krzysztof; Alper, Joseph; Grodzinski, Piotr

    2010-06-01

    Nanotechnology offers the potential for new approaches to detecting, treating, and preventing cancer. To determine the current status of the cancer nanotechnology field and the optimal path forward, the National Cancer Institute's Alliance for Nanotechnology in Cancer held three strategic workshops, covering the areas of in vitro diagnostics and prevention, therapy and post-treatment, and in vivo diagnosis and imaging. At each of these meetings, a wide range of experts from academia, industry, the nonprofit sector, and the U.S. government discussed opportunities in the field of cancer nanotechnology and barriers to its implementation. PMID:20460532

  14. Reflections on strategic nurse leadership.

    PubMed

    White, Jean

    2012-10-01

    This paper sets out some personal reflections by the Chief Nursing Officer for Wales on the challenges facing nurses and midwives as they undertake strategic leadership roles in NHS organisations. The paper looks at the national approach taken in Wales where behavioural competencies for executive nurse directors have been implemented. It considers the implications of the breadth of responsibilities executive nurse directors have and the importance of developing and supporting middle grade nurse managers and clinical directors. It concludes by looking at who is responsible for care within an organisation.

  15. Carlsbad Area Office strategic plan

    SciTech Connect

    1995-10-01

    This edition of the Carlsbad Area Office Strategic Plan captures the U.S. Department of Energy`s new focus, and supercedes the edition issued previously in 1995. This revision reflects a revised strategy designed to demonstrate compliance with environmental regulations earlier than the previous course of action; and a focus on the selected combination of scientific investigations, engineered alternatives, and waste acceptance criteria for supporting the compliance applications. An overview of operations and historical aspects of the Waste Isolation Pilot Plant near Carlsbad, New Mexico is presented.

  16. Strategic defense initiative: Folly or future

    SciTech Connect

    Haley, P.E.; Merritt, J.

    1986-01-01

    This collection of analyses is a guide through the maze of claims and criticisms about ''Star Wars,'' the controversial effort of the Reagan administration to reorient United States nuclear strategy to strategic defense. The text starts with an introduction by the editors followed by individual chapters outlining the strategic defense initiative as originally conceived and subsequently modified by the Reagan administration; the arguments for and against the plan's strategic and technical feasibility; and assessments of the harmful and constructive effects of strategic defense on U.S.-Soviet and U.S.-allied relations.

  17. 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.

  18. Strategic Petroleum Reserve annual report for calendar year 1998

    SciTech Connect

    1998-12-31

    The Strategic Petroleum Reserve was established in 1975 as an emergency response to the 1973 Arab oil embargo. It is authorized by the Energy Policy and Conservation Act (EPCA), and by the comprehensive energy plans of all Administrations since 1975, in recognition of the long-term dependence of the US on imported crude oil and petroleum products. Section 165 of EPCA requires the Secretary of Energy to submit an Annual Report to the President and the Congress. On May 13, 1998, the Department published a Statement of Administration Policy which reaffirmed its commitment to maintain a Government-owned and controlled, centrally located Strategic Petroleum Reserve of crude oil. The Reserve is to be used solely for responding to the types of severe oil supply interruptions presently contemplated in EPCA. Over the past twenty years, the Reserve has grown as large as 592 million barrels--a peak reached in 1994. From 1994 to 1996, nearly 28 million barrels were sold to raise revenues for the U S Treasury. As of December 31, 1998, the crude oil inventory was 561,108,127 barrels which equated to 60 days of net oil imports during 1998. The US now relies on a combination of both the Reserve and private stocks to meet its oil storage obligations to the International Energy Agency.

  19. 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: Setting…

  20. Visitor center museum display, detail of diorama booth with raised ...

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

    Visitor center museum display, detail of diorama booth with raised step; door to electrical door panel at left - Wright Brothers National Memorial Visitor Center, Highway 158, Kill Devil Hills, Dare County, NC