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Sample records for raising cre strategic

  1. Raising financing through strategic timing

    NASA Astrophysics Data System (ADS)

    Maine, Elicia; Thomas, V. J.

    2017-02-01

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

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

    ERIC Educational Resources Information Center

    Griswold, John S.; Jarvis, William F.

    2012-01-01

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

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

    DTIC Science & Technology

    2012-01-01

    Research Fellow in the Center for Strategic Research ( CSR ), Institute for National Strategic Studies, at the National Defense University. Michael...Kofman is a Program Manager and Researcher in CSR . Key Points ◆◆ With the New Strategic Arms Re- duction Treaty in place, the United States and Russia...country can be confident of its ability to field adequate defenses against attack by the other in any of these strategic domains. Neither one can

  4. 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…

  5. 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…

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

    PubMed

    Chapon, Pierre-Marie

    2011-06-01

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

  7. 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…

  8. 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.

  9. Foxn1[Cre] Expression in the Male Germline

    PubMed Central

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

    2016-01-01

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

  10. Carbapenem-Resistant Enterobacteriaceae (CRE) Infection

    MedlinePlus

    ... about VAP Diseases and Organisms Acinetobacter Burkholderia cepacia Clostridium difficile Patients Clinicians FAQs about C. difficile for ... Facilities/Settings State Health Departments Tracking C. difficile Clostridium Sordellii Carbapenem-resistant Enterobacteriaceae (CRE) Tracking CRE Interim ...

  11. Unmodified Cre recombinase crosses the membrane

    PubMed Central

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

    2002-01-01

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

  12. Skeletal defects in Osterix-Cre transgenic mice.

    PubMed

    Huang, Wei; Olsen, Bjorn R

    2015-02-01

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

  13. 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

  14. Raising consciousness.

    PubMed

    Hirsch, Joy

    2005-05-01

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

  15. Raising Feminists.

    ERIC Educational Resources Information Center

    Katz, Phyllis A.

    1996-01-01

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

  16. 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.

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

    PubMed Central

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

    2012-01-01

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

  18. 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

  19. 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

  20. Inducible control of gene expression with destabilized Cre

    PubMed Central

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

    2014-01-01

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

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

    PubMed Central

    Bhattacharyya, Samadrita; Bhakta, Minoti

    2017-01-01

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

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

    PubMed Central

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

    2013-01-01

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

  3. Cre-inducible site-specific recombination in zebrafish oligodendrocytes.

    PubMed

    Pinzon-Olejua, Alejandro; Welte, Cornelia; Chekuru, Avinash; Bosak, Viktoria; Brand, Michael; Hans, Stefan; Stuermer, Claudia A O

    2017-01-01

    The conditional Cre/lox system has recently emerged as a valuable tool for studies on both embryonic and adult Zebrafish. Temporal control and site-specific recombination are achieved by using the ligand-inducible CreER(T2) and administration of the drug tamoxifen (TAM) or its active metabolite, 4-Hydroxytamoxifen (4-OHT). Here we report the generation of a transgenic Zebrafish line, which expresses an mCherry-tagged variant of CreER(T2) under the control of the myelin basic protein a (mbpa) promoter. Our analysis shows that larval and adult expression of the transgene recapitulates the endogenous mbpa expression pattern in oligodendrocytes. Furthermore, combination with a Cre-dependent EGFP reporter results in EGFP-expressing oligodendrocytes in the spinal cord, brain, and optic nerve in TAM- or 4-OHT-treated larvae and 4-month-old fish, but not in untreated controls. The transgenic Zebrafish line Tg(mbpa:mCherry-T2A-CreER(T2) ) elicits CreER(T2) expression specifically in myelinating glia cells. Cre-inducible targeted recombination of genes in oligodendrocytes will be useful to elucidate cellular and molecular mechanisms of myelination in vivo during development (myelination) and regeneration (remyelination) after injury to the central nervous system (CNS). It will also allow targeted expression and overexpression of genes of interest (transgenes) in oligodendrocytes at defined developmental and adult stages. Developmental Dynamics 246:41-49, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

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

    PubMed

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

    2005-09-01

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

  5. Dwarfism in homozygous Agc1(CreERT) mice is associated with decreased expression of aggrecan.

    PubMed

    Rashid, Harunur; Chen, Haiyan; Hassan, Quamarul; Javed, Amjad

    2017-09-18

    Aggrecan (Acan), a large proteoglycan is abundantly expressed in cartilage tissue. Disruption of Acan gene causes dwarfism and perinatal lethality of homozygous mice. Because of sustained expression of Acan in the growth plate and articular cartilage, Agc(Cre) model has been developed for the regulated ablation of target gene in chondrocytes. In this model, the IRES-CreERT-Neo-pgk transgene is knocked-in the 3'UTR of the Acan gene. We consistently noticed variable weight and size among the Agc(Cre) littermates, prompting us to examine the cause of this phenotype. Wild-type, Cre-heterozygous (Agc(+/Cre) ), and Cre-homozygous (Agc(Cre/Cre) ) littermates were indistinguishable at birth. However, by 1-month, Agc(Cre/Cre) mice showed a significant reduction in body weight (18-27%) and body length (19-22%). Low body weight and dwarfism was sustained through adulthood and occurred in both genders. Compared with wild-type and Agc(+/Cre) littermates, long bones and vertebrae were shorter in Agc(Cre/Cre) mice. Histological analysis of Agc(Cre/Cre) mice revealed a significant reduction in the length of the growth plate and the thickness of articular cartilage. The amount of proteoglycan deposited in the cartilage of Agc(Cre/Cre) mice was nearly half of the WT littermates. Analysis of gene expression indicates impaired differentiation of chondrocyte in hyaline cartilage of Agc(Cre/Cre) mice. Notably, both Acan mRNA and protein was reduced by 50% in Agc(Cre/Cre) mice. A strong correlation was noted between the level of Acan mRNA and the body length. Importantly, Agc(+/Cre) mice showed no overt skeletal phenotype. Thus to avoid misinterpretation of data, only the Agc(+/Cre) mice should be used for conditional deletion of a target gene in the cartilage tissue. © 2017 Wiley Periodicals, Inc.

  6. 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…

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

    PubMed

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

    2014-04-01

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

  8. Col2-Cre and tamoxifen-inducible Col2-CreER target different cell populations in the knee joint

    PubMed Central

    Nagao, Masashi; Cheong, Chan Wook; Olsen, Bjorn

    2015-01-01

    Objective Collagen type 2 (Col2)-Cre or tamoxifen-inducible Col2-CreER transgenic mouse lines have been used for studies to explore the cellular and molecular pathogenesis of osteoarthritis (OA). The purpose of this study is to investigate whether the targeted cells are the same or different in the two mouse lines. Methods We crossed tamoxifen inducible Col2-CreER and Col2-Cre mice with Rosa tdTomato reporter mice and analyzed the labeling patterns at different time points. Results In the Col2-CreER mice, 90.8 [95% confidence interval (CI) (88.3, 93.2)] and 82.8 (77.4, 88.3) % of the articular surface cells are Tomato positive when tamoxifen was administered at 2 and 2.5 weeks of age and strong activity was observed even 4.5 months after injection. However, 46.0 (32.8, 59.1) and 22.2 (11.7, 32.6) % of the surface cells were Tomato positive when tamoxifen was administered at 3 and 4 weeks of age, respectively. Little to no Tomato activity in the articular surface cells was observed when tamoxifen was administered at 8 weeks of age. At any stage of tamoxifen injection, the Tomato activity was detected in growth plate and epiphyseal bone in addition to articular chondrocytes, but little in endothelium and not in the synovium and ligament. In contrast, the targeted tissues in the Col2-Cre mouse line were articular cartilage, growth plate, meniscus, endosteum, ligament, bone and synovium. Conclusions This study demonstrates that the pattern of targeted cells in the inducible Col2-CreER mice are partially overlapping with but different from that of targeted cells in Col2-Cre mice and the pattern varies dependent on when tamoxifen is administered. PMID:26256767

  9. Relationship of the CreBC two-component regulatory system and inner membrane protein CreD with swimming motility in Stenotrophomonas maltophilia.

    PubMed

    Huang, Hsin-Hui; Chen, Wei-Ching; Lin, Cheng-Wen; Lin, Yi-Tsung; Ning, Hsiao-Chen; Chang, Yi-Chih; Yang, Tsuey-Ching

    2017-01-01

    The CreBC two-component system (TCS) is a conserved regulatory system found in Escherichia coli, Aeromonas spp., Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. In this study, we determined how CreBC TCS regulates secreted protease activities and swimming motility using creB, creC, and creBC in-frame deletion mutants (KJΔCreB, KJΔCreC, and KJΔBC) of S. maltophilia KJ. Compared to wild-type KJ, KJΔCreB had a comparable secreted protease activity; however, the secreted protease activities were obviously reduced in KJΔCreC and KJΔBC, suggesting that CreC works together with another unidentified response regulator (not CreB) to regulate secreted protease activity. Single gene inactivation of creB or creC resulted in mutants with an enhanced swimming motility, and this phenotype was exacerbated in a double mutant KJΔBC. To elucidate the underlying mechanism responsible for the ΔcreBC-mediated swimming enhancement, flagella morphology observation, RNA-seq based transcriptome assay, qRT-PCR, and membrane integrity and potential assessment were performed. Flagella morphological observation ruled out the possibility that swimming enhancement was due to altered flagella morphology. CreBC inactivation upregulated the expression of creD and flagella-associated genes encoding the basal body- and motor-associated proteins. Furthermore, KJΔBC had an increased membrane susceptibility to Triton X-100 and CreD upregulation in KJΔBC partially alleviated the compromise of membrane integrity. The impact of creBC TCS on bacterial membrane potential was assessed by carbonyl cyanide m-chlorophenyl hydrazine (CCCP50) concentration at which 50% of bacterial swimming is inhibited. CCCP50 of wild-type KJ increased when creBC was deleted, indicating an association between the higher membrane potential of KJΔBC cells and enhanced motility. Upregulation of the basal body- and motor-associated genes of flagella in KJΔBC cells may explain the increased membrane potential

  10. Strategic Technology

    DTIC Science & Technology

    2012-03-11

    of massive multi-player gaming environments to facilitate expert education, collaboration, and experimentation. Finally, it recommends a focus on red ... team analysis and experimentation as a means of minimizing strategic surprise from technological sources. To fully explore potential strategic

  11. Novel ROSA26 Cre-reporter knock-in C57BL/6N mice exhibiting green emission before and red emission after Cre-mediated recombination.

    PubMed

    Hasegawa, Yoshikazu; Daitoku, Yoko; Sekiguchi, Keito; Tanimoto, Yoko; Mizuno-Iijima, Saori; Mizuno, Seiya; Kajiwara, Noriko; Ema, Masatsugu; Miwa, Yoshihiro; Mekada, Kazuyuki; Yoshiki, Atsushi; Takahashi, Satoru; Sugiyama, Fumihiro; Yagami, Ken-ichi

    2013-01-01

    The Cre/loxP system is a strategy for controlling temporal and/or spatial gene expression through genome alteration in mice. As successful Cre/loxP genome alteration depends on Cre-driver mice, Cre-reporter mice are essential for validation of Cre gene expression in vivo. In most Cre-reporter mouse strains, although the presence of reporter product indicates the expression of Cre recombinase, it has remained unclear whether a lack of reporter signal indicates either no Cre recombinase expression or insufficient reporter gene promoter activity. We produced a novel ROSA26 knock-in Cre-reporter C57BL/6N strain exhibiting green emission before and red after Cre-mediated recombination, designated as strain R26GRR. Ubiquitous green fluorescence and no red fluorescence were observed in R26GRR mice. To investigate the activation of tdsRed, EGFP-excised R26GRR, R26RR, mice were produced through the crossing of C57BL/6N mice with R26GRR/Ayu1-Cre F1 mice. R26RR mice showed extraordinarily strong red fluorescence in almost all tissues examined, suggesting ubiquitous activation of the second reporter in all tissues after Cre/loxP recombination. Moreover, endothelial cell lineage and pancreatic islet-specific expression of red fluorescence were detected in R26GRR/Tie2-Cre F1 mice and R26GRR /Ins1-Cre F1 mice, respectively. These results indicated that R26GRR mice are a useful novel Cre-reporter mouse strain. In addition, R26GRR mice with a pure C57BL/6N background represent a valuable source of green-to-red photoconvertible cells following Cre/loxP recombination for application in transplantation studies. The R26GRR mouse strain will be available from RIKEN BioResource Center (http://www.brc.riken.jp/lab/animal/en/).

  12. 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

  13. Thinking Strategically.

    ERIC Educational Resources Information Center

    Jeffress, Conway

    2000-01-01

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

  14. Cre Levels Limit Packaging Signal Excision Efficiency in the Cre/loxP Helper-Dependent Adenoviral Vector System

    PubMed Central

    Ng, Philip; Evelegh, Carole; Cummings, Derek; Graham, Frank L.

    2002-01-01

    Helper-dependent (HD) adenovirus vectors devoid of all viral coding sequences have a large cloning capacity and provide long-term transgene expression in vivo with negligible toxicity, making them attractive vectors for gene therapy. Currently, the most efficient means of producing HD vectors involves coinfecting 293 cells expressing Cre with the HD vector and a helper virus bearing a packaging signal flanked by loxP sites. Cre-mediated packaging signal excision renders the helper virus genome unpackageable but still able to replicate and provide helper functions for HD vector propagation. Typically, helper virus contamination is ≤1% pre- and ≤0.1% postpurification by CsCl banding. While these contamination levels are low, further reduction is desirable. However, this objective has not been realized since the Cre/loxP system was first developed. This lack of progress is due, at least in part, to our lack of understanding of the origins of the contaminating helper virus, thus rendering its reduction or elimination difficult to achieve. This study was designed to investigate the possible sources of contaminating helper virus persisting during HD vector amplification. The results revealed that Cre is limiting in helper virus-infected Cre-expressing 293 cells, thereby permitting helper viruses to escape packaging signal excision and propagate. The results of this study should provide a foundation for developing rational strategies to further reduce or possibly eliminate the contaminating helper virus. PMID:11932383

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  17. CRISPR/Cas9-mediated targeting of the Rosa26 locus produces Cre reporter rat strains for monitoring Cre-loxP-mediated lineage tracing.

    PubMed

    Ma, Yuanwu; Yu, Lei; Pan, Shuo; Gao, Shan; Chen, Wei; Zhang, Xu; Dong, Wei; Li, Jing; Zhou, Rui; Huang, Lan; Han, Yunlin; Bai, Lin; Zhang, Li; Zhang, Lianfeng

    2017-10-01

    The rat is an important laboratory animal for physiological, toxicological and pharmacological studies. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) is a simple and efficient tool to generate precise genetic modifications in rats, which will promote the accumulation of genetic resources and enable more precise studies of gene function. To monitor Cre-loxP-mediated excision in vivo, we generated a Cre reporter rat strain (Rosa26-imCherry) by knockin of a Cre reporter cassette at the Rosa26 locus using CRISPR/Cas9. Rosa26-imCherry rats exhibited inducible expression of the mCherry cassette (imCherry) using the Cre-loxP system, whereas normal rats exhibited ubiquitous expression of eGFP but not mCherry in the whole body. Injection of adeno-associated virus serotype 9-Cre into the hippocampus and skeletal muscle resulted in mCherry expression in virus-infected cells. Cre-loxP-mediated mCherry expression was then evaluated by crossing Rosa26-imCherry rats with transgenic rats ubiquitously expressing CAG-Cre, heart-specific α-MHC-Cre transgenic rats and liver-specific Alb-Cre knockin rats. Finally, using the established system the expression pattern of Cre driven by two endogenous gene promoters (Wfs1-Cre knockin rat, FabP2-Cre knockin rat) was traced. In summary, we demonstrated excision of the loxP-flanked allele in Rosa26-imCherry rats via activation of mCherry expression in the presence of Cre recombinase. This newly established Rosa26-imCherry rat strain represents a useful tool to facilitate Cre-expression pattern determination and tracing experiments. © 2017 Federation of European Biochemical Societies.

  18. Strategic weapons

    SciTech Connect

    Not Available

    1991-09-01

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

  19. Raising Children Who Refuse To Be Raised.

    ERIC Educational Resources Information Center

    Ziegler, Dave

    This book is about the struggle of raising children who work against your every effort as a parent or clinician to help them succeed in the world. Specifically, this book is about children who often do not respond to approaches offered in books and parent training programs for "normal" children. There are other books that address individual…

  20. A photoactivatable Cre-loxP recombination system for optogenetic genome engineering.

    PubMed

    Kawano, Fuun; Okazaki, Risako; Yazawa, Masayuki; Sato, Moritoshi

    2016-12-01

    Genome engineering techniques represented by the Cre-loxP recombination system have been used extensively for biomedical research. However, powerful and useful techniques for genome engineering that have high spatiotemporal precision remain elusive. Here we develop a highly efficient photoactivatable Cre recombinase (PA-Cre) to optogenetically control genome engineering in vivo. PA-Cre is based on the reassembly of split Cre fragments by light-inducible dimerization of the Magnet system. PA-Cre enables sharp induction (up to 320-fold) of DNA recombination and is efficiently activated even by low-intensity illumination (∼0.04 W m(-2)) or short periods of pulsed illumination (∼30 s). We demonstrate that PA-Cre allows for efficient DNA recombination in an internal organ of living mice through noninvasive external illumination using a LED light source. The present PA-Cre provides a powerful tool to greatly facilitate optogenetic genome engineering in vivo.

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

    PubMed Central

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

    2001-01-01

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

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

    PubMed

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

    2005-02-01

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2014-11-01

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

  5. Strategic Genius

    DTIC Science & Technology

    2015-05-21

    Theory: Modern , Symbolic, and Postmodern Perspectives (Oxford: Oxford University Press, 2013), 185–188. 10 status quo solutions remain...York: Oxford University Press, 1999. Hatch, Mary Jo. Organization Theory: Modern , Symbolic, and Postmodern Perspectives. Oxford: Oxford University...elusive strategic genius. What traits make a strategic genius? Several modern theorists examine how such an individual might think and some offer how

  6. Raising a Fit Preschooler

    MedlinePlus

    ... 10 Tips for Parents Healthy Habits for TV, Video Games, and the Internet Raising a Fit Preschooler KidsHealth > For Parents > Raising a Fit Preschooler Print A A A What's in this article? Helping Kids Learn New Skills How Much Activity Is Enough? Structured Play Unstructured ...

  7. 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,"…

  8. 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,"…

  9. 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

  10. Mouse opsin promoter-directed Cre recombinase expression in transgenic mice.

    PubMed

    Le, Yun-Zheng; Zheng, Lixin; Zheng, Wei; Ash, John D; Agbaga, Martin-Paul; Zhu, Meili; Anderson, Robert E

    2006-04-18

    Gene inactivation with homologous recombination in mice is a widely used tool to study gene function. However, many proteins play essential roles in a number of tissues and germline gene inactivation often results in embryonic lethality. To overcome this limitation and to dissect the functions of essential genes beyond embryonic development, we generated mouse rod opsin promoter-controlled cre transgenic mice with a goal of obtaining transgenic lines with a range of Cre activity in rod photoreceptors. Transgenic mice expressing Cre recombinase directed by a long or short mouse opsin promoter were generated. Candidate Cre-expressing lines were identified with RT-PCR and Western blot analysis. Potentially useful Cre-expressing lines were characterized further with immunohistochemistry, PCR, and functional analysis using a Cre-activatable lacZ reporter mouse strain (R26R) to determine temporal and spatial patterns of Cre expression. Retinal function and morphology in these mouse lines were analyzed with electroretinography (ERG) and light microscopy of hematoxylin and eosin stained retinal sections. Transgenic mice expressing Cre in rod photoreceptors were generated. Characterization of candidate photoreceptor-specific Cre mice using immunohistochemistry and functional assays demonstrated that an efficient Cre-mediated recombination occurred in rod photoreceptor cells in one mouse line and a mosaic Cre-mediated recombination occurred in rod photoreceptors and rod bipolar cells in another mouse line. Further analysis of these mice with ERG and morphological examination suggested that the retinas of eight-month-old adults were normal. We have generated transgenic mice expressing Cre recombinase in rod photoreceptors. One transgenic mouse line was capable of carrying out efficient Cre-mediated recombination in rod photoreceptors. Another transgenic mouse line was capable of carrying out mosaic Cre-mediated recombination in rod photoreceptors and bipolar cells across the

  11. SPC-Cre-ERT2 Transgenic Mouse for Temporal Gene Deletion in Alveolar Epithelial Cells

    PubMed Central

    Gui, Yao-Song; Wang, Lianmei; Tian, Xinlun; Feng, Ruie; Ma, Aiping; Cai, Baiqiang; Zhang, Hongbing; Xu, Kai-Feng

    2012-01-01

    Although several Cre-loxP-based gene knockout mouse models have been generated for the study of gene function in alveolar epithelia in the lung, their applications are still limited. In this study, we developed a SPC-Cre-ERT2 mouse model, in which a tamoxifen-inducible Cre recombinase (Cre-ERT2) is under the control of the human surfactant protein C (SPC) promoter. The specificity and efficiency of Cre-ERT2 activity was first evaluated by crossing SPC-Cre-ERT2 mouse with ROSA26R mouse, a β-galactosidase reporter strain. We found that Cre-ERT2 was expressed in 30.7% type II alveolar epithelial cells of SPC-Cre-ERT2/ROSA26R mouse lung tissues in the presence of tamoxifen. We then tested the tamoxifen-inducible recombinase activity of Cre-ERT2 in a mouse strain bearing TSC1 conditional knockout alleles (TSC1fx/fx). TSC1 deletion was detected in the lungs of tamoxifen treated SPC-Cre-ERT2/TSC1fx/fx mice. Therefore this SPC-Cre-ERT2 mouse model may be a valuable tool to investigate functions of genes in lung development, physiology and disease. PMID:23049940

  12. SPC-Cre-ERT2 transgenic mouse for temporal gene deletion in alveolar epithelial cells.

    PubMed

    Gui, Yao-Song; Wang, Lianmei; Tian, Xinlun; Feng, Ruie; Ma, Aiping; Cai, Baiqiang; Zhang, Hongbing; Xu, Kai-Feng

    2012-01-01

    Although several Cre-loxP-based gene knockout mouse models have been generated for the study of gene function in alveolar epithelia in the lung, their applications are still limited. In this study, we developed a SPC-Cre-ER(T2) mouse model, in which a tamoxifen-inducible Cre recombinase (Cre-ER(T2)) is under the control of the human surfactant protein C (SPC) promoter. The specificity and efficiency of Cre-ER(T2) activity was first evaluated by crossing SPC-Cre-ER(T2) mouse with ROSA26R mouse, a β-galactosidase reporter strain. We found that Cre-ER(T2) was expressed in 30.7% type II alveolar epithelial cells of SPC-Cre-ER(T2)/ROSA26R mouse lung tissues in the presence of tamoxifen. We then tested the tamoxifen-inducible recombinase activity of Cre-ER(T2) in a mouse strain bearing TSC1 conditional knockout alleles (TSC1(fx/fx)). TSC1 deletion was detected in the lungs of tamoxifen treated SPC-Cre-ER(T2)/TSC1(fx/fx) mice. Therefore this SPC-Cre-ER(T2) mouse model may be a valuable tool to investigate functions of genes in lung development, physiology and disease.

  13. Nrl-Cre transgenic mouse mediates loxP recombination in developing rod photoreceptors

    PubMed Central

    Brightman, Diana S.; Razafsky, David; Potter, Chloe; Hodzic, Didier; Chen, Shiming

    2016-01-01

    The developing mouse retina is a tractable model for studying neurogenesis and differentiation. Although transgenic Cre mouse lines exist to mediate conditional genetic manipulations in developing mouse retinas, none of them act specifically in early developing rods. For conditional genetic manipulations of developing retinas, we created a Nrl-Cre mouse line in which the Nrl promoter drives expression of Cre in rod precursors. Our results show that Nrl-Cre expression is specific to the retina where it drives rod-specific recombination with a temporal pattern similar to endogenous Nrl expression during retinal development. This Nrl-Cre transgene does not negatively impact retinal structure and function. Taken together, our data suggest that the Nrl-Cre mouse line is a valuable tool to drive Cre-mediated recombination specifically in developing rods. PMID:26789558

  14. Nrl-Cre transgenic mouse mediates loxP recombination in developing rod photoreceptors.

    PubMed

    Brightman, Diana S; Razafsky, David; Potter, Chloe; Hodzic, Didier; Chen, Shiming

    2016-03-01

    The developing mouse retina is a tractable model for studying neurogenesis and differentiation. Although transgenic Cre mouse lines exist to mediate conditional genetic manipulations in developing mouse retinas, none of them act specifically in early developing rods. For conditional genetic manipulations of developing retinas, a Nrl-Cre mouse line in which the Nrl promoter drives expression of Cre in rod precursors was created. The results showed that Nrl-Cre expression was specific to the retina where it drives rod-specific recombination with a temporal pattern similar to endogenous Nrl expression during retinal development. This Nrl-Cre transgene does not negatively impact retinal structure and function. Taken together, the data suggested that the Nrl-Cre mouse line was a valuable tool to drive Cre-mediated recombination specifically in developing rods. © 2016 Wiley Periodicals, Inc.

  15. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain.

    PubMed

    Madisen, Linda; Zwingman, Theresa A; Sunkin, Susan M; Oh, Seung Wook; Zariwala, Hatim A; Gu, Hong; Ng, Lydia L; Palmiter, Richard D; Hawrylycz, Michael J; Jones, Allan R; Lein, Ed S; Zeng, Hongkui

    2010-01-01

    The Cre/lox system is widely used in mice to achieve cell-type-specific gene expression. However, a strong and universally responding system to express genes under Cre control is still lacking. We have generated a set of Cre reporter mice with strong, ubiquitous expression of fluorescent proteins of different spectra. The robust native fluorescence of these reporters enables direct visualization of fine dendritic structures and axonal projections of the labeled neurons, which is useful in mapping neuronal circuitry, imaging and tracking specific cell populations in vivo. Using these reporters and a high-throughput in situ hybridization platform, we are systematically profiling Cre-directed gene expression throughout the mouse brain in several Cre-driver lines, including new Cre lines targeting different cell types in the cortex. Our expression data are displayed in a public online database to help researchers assess the utility of various Cre-driver lines for cell-type-specific genetic manipulation.

  16. Strategic Materials

    DTIC Science & Technology

    2006-01-01

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

  17. Raising Children Again

    MedlinePlus

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

  18. 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)

  19. 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)

  20. Expedition 8 Flag Raising

    NASA Image and Video Library

    2003-10-14

    The flags of the United States, Russia, Spain and Kazakhstan fly outside the Cosmonaut Hotel, Tuesday, Oct. 14, 2003, in Baikonur, Kazakhstan following a traditional flag-raising ceremony. Photo Credit (NASA/Bill Ingalls)

  1. 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)

  2. 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)

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

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

  4. 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.

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

    PubMed Central

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

    2011-01-01

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

  6. A New Way of Thinking About Strategic Sourcing

    DTIC Science & Technology

    2016-05-17

    7 Army Sustainment July–August 2015 COM M ENTARY Strategic sourcing has been cre-ating efficiencies , improving the quality of services, and re...additional training and operations? What if the Army could eliminate redundant contracts and truly implement the intent of portfolio management for...Kroul and Jerry E. Jastrab By using an Army Sustainment Command strategy of co-locating portfolio managers, the Army could more quickly realize its

  7. Arity Raising in Manticore

    NASA Astrophysics Data System (ADS)

    Bergstrom, Lars; Reppy, John

    Compilers for polymorphic languages are required to treat values in programs in an abstract and generic way at the source level. The challenges of optimizing the boxing of raw values, flattening of argument tuples, and raising the arity of functions that handle complex structures to reduce memory usage are old ones, but take on newfound import with processors that have twice as many registers. We present a novel strategy that uses both control-flow and type information to provide an arity raising implementation addressing these problems. This strategy is conservative - no matter the execution path, the transformed program will not perform extra operations.

  8. Cre-Mediated Recombination in Rhombic Lip Derivatives

    PubMed Central

    Fünfschilling, Ursula; Reichardt, Louis F.

    2009-01-01

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

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

    PubMed

    Yarmolinsky, Michael; Hoess, Ronald

    2015-11-01

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

  10. Raising Responsible Teenagers.

    ERIC Educational Resources Information Center

    Myers, Bob

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

  11. Raising Better Boys.

    ERIC Educational Resources Information Center

    Canada, Geoffrey

    2000-01-01

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

  12. Raising Better Boys.

    ERIC Educational Resources Information Center

    Canada, Geoffrey

    2000-01-01

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

  13. Expedition 8 Flag Raising

    NASA Image and Video Library

    2003-10-14

    Expedition 8 Commander and NASA Science Officer Michael Foale, left, and European Space Agency astronaut Pedro Duque of Spain participate in the traditional raising of the flags at the Cosmonaut Hotel, Tuesday, Oct. 14, 2003, in Baikonur, Kazakhstan. Photo Credit (NASA/Bill Ingalls)

  14. Raise Funds, Not Hackles.

    ERIC Educational Resources Information Center

    Zaidel, Lisa Brusman

    1996-01-01

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

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

    PubMed

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

    2016-09-01

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

  16. 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)

  17. Strategic HRD.

    ERIC Educational Resources Information Center

    1997

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

  18. 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;…

  19. Strategic HRD.

    ERIC Educational Resources Information Center

    1997

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

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

    PubMed

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

    2002-03-01

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

  1. 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.

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

    PubMed

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

    2014-01-01

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

  3. Removal of Carbapenem-Resistant Enterobacteriaceae (CRE) from blood by heparin-functional hemoperfusion media.

    PubMed

    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.

  4. Generation and characterization of an Advillin-Cre driver mouse line

    PubMed Central

    2011-01-01

    Progress in the somatosensory field has been restricted by the limited number of genetic tools available to study gene function in peripheral sensory neurons. Here we generated a Cre-driver mouse line that expresses Cre-recombinase from the locus of the sensory neuron specific gene Advillin. These mice displayed almost exclusive Cre-mediated recombination in all peripheral sensory neurons. As such, the Advillin-Cre-driver line will be a powerful tool for targeting peripheral neurons in future investigations. PMID:21906401

  5. Strategic Supply

    DTIC Science & Technology

    2006-01-01

    leaders as Sears, Limited Brands, DHL, Circuit City, Cingular, Nestle and IKEA (Manugistics, 2006). The Strategic Supply Chain Industry Study Group...inventory turns have increased. Other global customers have also reaped the benefits of the Manugistics software. IKEA , Sweden’s retail icon...turned to Manugistics after a mid-1990s ERP implementation failed to fix their forecasting problems, which gave way to fluctuating inventory levels. IKEA

  6. A tetracycline- inducible and skeletal muscle specific Cre recombinase transgenic mouse

    PubMed Central

    Rao, Pengcheng; Monks, D. Ashley

    2009-01-01

    We have generated a transgenic mouse that expresses Cre recombinase only in skeletal muscle and only following tetracycline-treatment. This spatiotemporal specificity is achieved using 2 transgenes. The first transgene uses the human skeletal actin (HSA) promoter to drive expression of the reverse tetracycline-controlled transactivator (rtTA). The second transgene uses a tetracycline responsive promoter to drive expression of Cre recombinase. We monitored transgene expression in these mice by crossing them with ROSA26 loxP-LacZ reporter mice, which express β–galactosidase when activated by Cre. We find that expression of this transgene is only detectable within skeletal muscle and that Cre expression in the absence of tetracycline is negligible. Cre is readily induced in this model with tetracycline analogs at a range of embryonic and postnatal ages and in a pattern consistent with other HSA transgenic mice. This mouse improves upon existing transgenic mice in which skeletal muscle Cre is expressed throughout development by allowing Cre expression to begin at later developmental stages. This temporal control of transgene expression has several applications, including overcoming embryonic or perinatal lethality due to transgene expression. This mouse is especially suited for studies of steroid hormone action, as it uses tetracycline, rather than tamoxifen to activate Cre expression. In summary, we find that this transgenic induction system is suitable for studies of gene function in the context of hormonal regulation of skeletal muscle and/or interactions between muscle and motoneurons in mice. PMID:19263419

  7. A tetracycline-inducible and skeletal muscle-specific Cre recombinase transgenic mouse.

    PubMed

    Rao, Pengcheng; Monks, D Ashley

    2009-05-01

    We have generated a transgenic mouse that expresses Cre recombinase only in skeletal muscle and only following tetracycline treatment. This spatiotemporal specificity is achieved using two transgenes. The first transgene uses the human skeletal actin (HSA) promoter to drive expression of the reverse tetracycline-controlled transactivator (rtTA). The second transgene uses a tetracycline responsive promoter to drive the expression of Cre recombinase. We monitored transgene expression in these mice by crossing them with ROSA26 loxP-LacZ reporter mice, which express beta-galactosidase when activated by Cre. We find that the expression of this transgene is only detectable within skeletal muscle and that Cre expression in the absence of tetracycline is negligible. Cre is readily induced in this model with tetracycline analogs at a range of embryonic and postnatal ages and in a pattern consistent with other HSA transgenic mice. This mouse improves upon existing transgenic mice in which skeletal muscle Cre is expressed throughout development by allowing Cre expression to begin at later developmental stages. This temporal control of transgene expression has several applications, including overcoming embryonic or perinatal lethality due to transgene expression. This mouse is especially suited for studies of steroid hormone action, as it uses tetracycline, rather than tamoxifen, to activate Cre expression. In summary, we find that this transgenic induction system is suitable for studies of gene function in the context of hormonal regulation of skeletal muscle or interactions between muscle and motoneurons in mice.

  8. 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

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

    PubMed

    Ding, Yuzhi; Li, Jianmin; Liu, Qinghuai

    2013-11-29

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed

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

    1998-09-01

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

  12. 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

  13. Measurements of DNA-loop formation via Cre-mediated recombination

    PubMed Central

    Shoura, Massa J.; Vetcher, Alexandre A.; Giovan, Stefan M.; Bardai, Farah; Bharadwaj, Anusha; Kesinger, Matthew R.; Levene, Stephen D.

    2012-01-01

    The Cre-recombination system has become an important tool for genetic manipulation of higher organisms and a model for site-specific DNA-recombination mechanisms employed by the λ-Int superfamily of recombinases. We report a novel quantitative approach for characterizing the probability of DNA-loop formation in solution using time-dependent ensemble Förster resonance energy transfer measurements of intra- and inter-molecular Cre-recombination kinetics. Our method uses an innovative technique for incorporating multiple covalent modifications at specific sites in covalently closed DNA. Because the mechanism of Cre recombinase does not conform to a simple kinetic scheme, we employ numerical methods to extract rate constants for fundamental steps that pertain to Cre-mediated loop closure. Cre recombination does not require accessory proteins, DNA supercoiling or particular metal-ion cofactors and is thus a highly flexible system for quantitatively analyzing DNA-loop formation in vitro and in vivo. PMID:22589415

  14. Fund-raising fiasco.

    PubMed

    1997-05-16

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

  15. 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.

  16. Strategic Petroleum Reserves

    SciTech Connect

    Not Available

    1989-01-01

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

  17. Strategic petroleum reserves

    SciTech Connect

    Not Available

    1989-04-01

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

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

    ERIC Educational Resources Information Center

    Brandt, Ron

    1991-01-01

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

  19. A mart-1::Cre transgenic line induces recombination in melanocytes and retinal pigment epithelium.

    PubMed

    Aydin, Iraz T; Beermann, Friedrich

    2011-05-01

    The number of transgenic mouse lines expressing Cre in either type of pigment cells (melanocytes and retinal pigment epithelium, RPE) is limited, and the available lines do not always offer sufficient specificity. In this study, we addressed this issue and we report on the generation of a MART-1::Cre BAC transgenic mouse line, in which the expression of Cre recombinase is controlled by regulatory elements of the pigment cell-specific gene MART-1 (mlana). When MART-1::Cre BAC transgenic mice were bred with the ROSA26-R reporter line, ß-galactosidase expression was observed in RPE from E12.5 onwards, and in melanocyte precursors from E17.5, indicating that the MART-1::Cre line provides Cre recombinase activity in pigment-producing cells rather than in a particular lineage. In addition, breeding of this mouse line to mice carrying a conditional allele of RBP-Jκ corroborated the reported phenotypes in both pigment cell lineages, inducing hair greying and microphthalmia. Our results thus suggest, that the MART-1::Cre line may serve as a novel and useful tool for functional studies in melanocytes and the RPE.genesis 49:403-409, 2011.

  20. 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.

  1. ATF-2 stimulates the human insulin promoter through the conserved CRE2 sequence.

    PubMed

    Hay, Colin W; Ferguson, Laura A; Docherty, Kevin

    2007-02-01

    The insulin promoter contains a number of dissimilar cis-acting regulatory elements that bind a range of tissue specific and ubiquitous transcription factors. Of the regulatory elements within the insulin promoter, the cyclic AMP responsive element (CRE) binds by far the most diverse array of transcription factors. Rodent insulin promoters have a single CRE site, whereas there are four CREs within the human insulin gene, of which CRE2 is the only one conserved between species. The aim of this study was to characterise the human CRE2 site and to investigate the effects of the two principal CRE-associated transcription factors; CREB-1 and ATF-2. Co-transfection of INS-1 pancreatic beta-cells with promoter constructs containing the human insulin gene promoter placed upstream of the firefly luciferase reporter gene and expression plasmids for ATF-2 or CREB-1 showed that ATF-2 stimulated transcriptional activity while CREB-1 elicited an inhibitory effect. Mutagenesis of CRE2 diminished the effect of ATF-2 but not that of CREB-1. ATF-2 was shown to bind to the CRE2 site by electrophoretic mobility shift assay and by chromatin immunoprecipitation, while siRNA mediated knockdown of ATF-2 diminished the stimulatory effects of cAMP related signalling on promoter activity. These results suggest that ATF-2 may be a key regulator of the human insulin promoter possibly stimulating activity in response to extracellular signals.

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

    PubMed Central

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

    2017-01-01

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

  3. [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.

  4. 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.

  5. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain

    PubMed Central

    Madisen, Linda; Zwingman, Theresa A.; Sunkin, Susan M.; Oh, Seung Wook; Zariwala, Hatim A.; Gu, Hong; Ng, Lydia L.; Palmiter, Richard D.; Hawrylycz, Michael J.; Jones, Allan R.; Lein, Ed S.; Zeng, Hongkui

    2009-01-01

    The Cre/lox system is widely used in mice to achieve cell-type-specific gene expression. However, a strong and universal responding system to express genes under Cre control is still lacking. We have generated a set of Cre reporter mice with strong, ubiquitous expression of fluorescent proteins of different spectra. The robust native fluorescence of these reporters enables direct visualization of fine dendritic structures and axonal projections of the labeled neurons, which is useful in mapping neuronal circuitry, imaging and tracking specific cell populations in vivo. Using these reporters and a high-throughput in situ hybridization platform, we are systematically profiling Cre-directed gene expression throughout the mouse brain in a number of Cre-driver lines, including novel Cre lines targeting different cell types in the cortex. Our expression data are displayed in a public online database to help researchers assess the utility of various Cre-driver lines for cell-type-specific genetic manipulation. PMID:20023653

  6. 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.

  7. Profiling of functional phosphodiesterase in mesangial cells using a CRE-SEAP-based reporting system.

    PubMed

    Zhu, Ying; Yao, Jian; Meng, Yiman; Kasai, Ayumi; Hiramatsu, Nobuhiko; Hayakawa, Kunihiro; Miida, Takashi; Takeda, Masayuki; Okada, Masahiko; Kitamura, Masanori

    2006-07-01

    1. Phosphodiesterases (PDEs) are critically implicated in the regulation of mesangial cell function, but profile of functional PDEs in mesangial cells is still unclear. In this study, we investigated roles of individual PDEs in the regulation of mesangial cell behavior by the cAMP pathway. 2. Reporter mesangial cells that express secreted alkaline phosphatase (SEAP) under the control of the cAMP response element (CRE) were exposed to selective PDE inhibitors in the presence or absence of cAMP, and activity of CRE, expression of CRE-regulated protein, mitogenesis and cell survival were examined. 3. Exposure of reporter cells to cAMP-elevating agents resulted in time- and concentration-dependent activation of CRE. Treatment of the cells with any PDE inhibitors alone did not induce CRE activation. Under stimulation with 8-bromo-cAMP or 8-bromo-cGMP, however, inhibitors of PDE2, PDE3, PDE4 and PDE5 enhanced activation of CRE. Inhibition of PDE1 or PDE6 did not affect the CRE activation. 4. Among different combinations tested, only inhibitors of PDE3 and PDE4 cooperatively increased the level of intracellular cAMP, activity of protein kinase A, activation of CRE, and CRE-regulated protein, connexin43. 5. Concomitant inhibition of PDE3 and PDE4 attenuated mitogen-induced activation of extracellular signal-regulated kinases and cell proliferation. Under serum deprivation, combinational inhibition of PDE3 and PDE4 exclusively caused activation of caspase-3 and apoptosis. 6. The present data elucidated that PDE3 and PDE4 play critical roles in the regulation of mesangial cell function. PDE3 and PDE4 were identified as the novel, antiapoptotic machinery that supports survival of mesangial cells.

  8. Profiling of functional phosphodiesterase in mesangial cells using a CRE-SEAP-based reporting system

    PubMed Central

    Zhu, Ying; Yao, Jian; Meng, Yiman; Kasai, Ayumi; Hiramatsu, Nobuhiko; Hayakawa, Kunihiro; Miida, Takashi; Takeda, Masayuki; Okada, Masahiko; Kitamura, Masanori

    2006-01-01

    Phosphodiesterases (PDEs) are critically implicated in the regulation of mesangial cell function, but profile of functional PDEs in mesangial cells is still unclear. In this study, we investigated roles of individual PDEs in the regulation of mesangial cell behavior by the cAMP pathway. Reporter mesangial cells that express secreted alkaline phosphatase (SEAP) under the control of the cAMP response element (CRE) were exposed to selective PDE inhibitors in the presence or absence of cAMP, and activity of CRE, expression of CRE-regulated protein, mitogenesis and cell survival were examined. Exposure of reporter cells to cAMP-elevating agents resulted in time- and concentration-dependent activation of CRE. Treatment of the cells with any PDE inhibitors alone did not induce CRE activation. Under stimulation with 8-bromo-cAMP or 8-bromo-cGMP, however, inhibitors of PDE2, PDE3, PDE4 and PDE5 enhanced activation of CRE. Inhibition of PDE1 or PDE6 did not affect the CRE activation. Among different combinations tested, only inhibitors of PDE3 and PDE4 cooperatively increased the level of intracellular cAMP, activity of protein kinase A, activation of CRE, and CRE-regulated protein, connexin43. Concomitant inhibition of PDE3 and PDE4 attenuated mitogen-induced activation of extracellular signal-regulated kinases and cell proliferation. Under serum deprivation, combinational inhibition of PDE3 and PDE4 exclusively caused activation of caspase-3 and apoptosis. The present data elucidated that PDE3 and PDE4 play critical roles in the regulation of mesangial cell function. PDE3 and PDE4 were identified as the novel, antiapoptotic machinery that supports survival of mesangial cells. PMID:16751794

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

    PubMed Central

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

    2014-01-01

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

  10. Cirrhosis Could Raise Stroke Risk

    MedlinePlus

    ... gov/news/fullstory_166267.html Cirrhosis Could Raise Stroke Risk Average yearly rate of the attacks doubled ... also raise an older person's odds for a stroke, a new study suggests. "In a nationally representative ...

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

    PubMed Central

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

    2006-01-01

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

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

    ERIC Educational Resources Information Center

    Strong, Bart

    2005-01-01

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

  13. Reliable Genetic Labeling of Adult-Born Dentate Granule Cells Using Ascl1CreERT2 and GlastCreERT2 Murine Lines

    PubMed Central

    Yang, Sung M.; Alvarez, Diego D.

    2015-01-01

    Newly generated dentate granule cells (GCs) are relevant for input discrimination in the adult hippocampus. Yet, their precise contribution to information processing remains unclear. To address this question, it is essential to develop approaches to precisely label entire cohorts of adult-born GCs. In this work, we used genetically modified mice to allow conditional expression of tdTomato (Tom) in adult-born GCs and characterized their development and functional integration. Ascl1CreERT2;CAGfloxStopTom and GlastCreERT2;CAGfloxStopTom mice resulted in indelible expression of Tom in adult neural stem cells and their lineage upon tamoxifen induction. Whole-cell recordings were performed to measure intrinsic excitability, firing behavior, and afferent excitatory connectivity. Developing GCs were also staged by the expression of early and late neuronal markers. The slow development of adult-born GCs characterized here is consistent with previous reports using retroviral approaches that have revealed that a mature phenotype is typically achieved after 6–8 weeks. Our findings demonstrate that Ascl1CreERT2 and GlastCreERT2 mouse lines enable simple and reliable labeling of adult-born GC lineages within restricted time windows. Therefore, these mice greatly facilitate tagging new neurons and manipulating their activity, required for understanding adult neurogenesis in the context of network remodeling, learning, and behavior. SIGNIFICANCE STATEMENT Our study shows that Ascl1CreERT2 and GlastCreERT2 mice lines can be used to label large cohorts of adult-born dentate granule cells with excellent time resolution. Neurons labeled in this manner display developmental and functional profiles that are in full agreement with previous findings using thymidine analogs and retroviral labeling, thus providing an alternative approach to tackle fundamental questions on circuit remodeling. Because of the massive neuronal targeting and the simplicity of this method, genetic labeling will

  14. Tg(Th-Cre)FI172Gsat (Th-Cre) defines neurons that are required for full hypercapnic and hypoxic reflexes.

    PubMed

    Sun, Jenny J; Ray, Russell S

    2017-08-15

    The catecholaminergic (CA) system has been implicated in many facets of breathing control and offers an important target to better comprehend the underlying etiologies of both developmental and adult respiratory pathophysiologies. Here, we used a noninvasive DREADD-based pharmacogenetic approach to acutely perturb Tg(Th-Cre)FI172Gsat (Th-Cre)-defined neurons in awake and unrestrained mice in an attempt to characterize CA function in breathing. We report that clozapine-N-oxide (CNO)-DREADD-mediated inhibition of Th-Cre-defined neurons results in blunted ventilatory responses under respiratory challenge. Under a hypercapnic challenge (5% CO2/21% O2/74% N2), perturbation of Th-Cre neurons results in reduced fR, [Formula: see text] and [Formula: see text] Under a hypoxic challenge (10% O2/90% N2), we saw reduced fR, [Formula: see text] and [Formula: see text], in addition to instability in both interbreath interval and tidal volume, resulting in a Cheyne-Stokes-like respiratory pattern. These findings demonstrate the necessity of Th-Cre-defined neurons for the hypercapnic and hypoxic ventilatory responses and breathing stability during hypoxia. However, given the expanded non-CA expression domains of the Tg(Th-Cre)FI172Gsat mouse line found in the brainstem, full phenotypic effect cannot be assigned solely to CA neurons. Nonetheless, this work identifies a key respiratory population that may lead to further insights into the circuitry that maintains respiratory stability in the face of homeostatic challenges. © 2017. Published by The Company of Biologists Ltd.

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

    MedlinePlus

    ... processing SSI file Error processing SSI file 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 ...

  16. Bigenic Cre/loxP, puΔtk conditional genetic ablation

    PubMed Central

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

    2004-01-01

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

  17. Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus.

    PubMed

    Wall, Nicholas R; Wickersham, Ian R; Cetin, Ali; De La Parra, Mauricio; Callaway, Edward M

    2010-12-14

    We describe a powerful system for revealing the direct monosynaptic inputs to specific cell types in Cre-expressing transgenic mice through the use of Cre-dependent helper virus and a modified rabies virus. We generated helper viruses that target gene expression to Cre-expressing cells, allowing us to control initial rabies virus infection and subsequent monosynaptic retrograde spread. Investigators can use this system to elucidate the connections onto a desired cell type in a high-throughput manner, limited only by the availability of Cre mouse lines. This method allows for identification of circuits that would be extremely tedious or impossible to study with other methods and can be used to build subcircuit maps of inputs onto many different types of cells within the same brain region. Furthermore, by expressing various transgenes from the rabies genome, this system also has the potential to allow manipulation of targeted neuronal circuits without perturbing neighboring cells.

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

    PubMed Central

    2012-01-01

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

  19. Raising the continental crust

    NASA Astrophysics Data System (ADS)

    Campbell, Ian H.; Davies, D. Rhodri

    2017-02-01

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

  20. 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

  1. Electrophysiological effects of CRE-1087 in guinea-pig ventricular muscles.

    PubMed Central

    Delpón, E.; Valenzuela, C.; Pérez, O.; Tamargo, J.

    1992-01-01

    1. The electrophysiological effects of CRE-1087, a new antiarrhythmic drug, were studied in guinea-pig papillary muscles. 2. CRE-1087 (10(-7) M-10(-4) M) produced a concentration-dependent decrease in the action potential amplitude and Vmax of the upstroke without altering the action potential duration or the resting membrane potential. 3. At 5 x 10(-6) M, CRE-1087 produced a 5.0 +/- 1.0% tonic Vmax block and this value was not modified at the different rates of stimulation tested. In the presence of CRE-1087, trains of stimuli at rates between 0.5 and 3 Hz led to an exponential decline in Vmax (frequency-dependent Vmax block) which augmented at higher rates of stimulation. At 2 Hz the onset kinetics of the frequency-dependent Vmax block was fitted by a monoexponential function being the K value 0.099 +/- 0.012 AP-1. The time constant for the recovery of Vmax from the frequency-dependent block was prolonged to 18.3 +/- 1.5 s. 4. CRE-1087 (5 x 10(-6) M) did not shift the membrane responsiveness curve. 5. CRE-1087 had no effect on the characteristics of the slow action potentials elicited by isoprenaline in ventricular fibres depolarized by 27 mM KCl, which suggested that CRE-1087 did not exhibit class IV (Ca antagonist) antiarrhythmic actions. 6. The slow onset and the slow offset kinetics of frequency-dependent Vmax block during repetitive activity suggested that in guinea-pig ventricular muscle fibres CRE-1087 exhibited class Ic antiarrhythmic actions. PMID:1422597

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2015-06-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

  6. 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

  7. 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.

  8. Cre-loxP-Mediated Recombination: General Principles and Experimental Considerations.

    PubMed

    McLellan, Micheal A; Rosenthal, Nadia A; Pinto, Alexander R

    2017-03-02

    The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. This has been largely due to the simplicity of its use and its adaptability to address diverse biological questions. While the use of the cre-loxP system is becoming increasingly widespread, in particular because of growing availability of conditional mouse mutants, many considerations need to be taken into account when utilizing the cre-loxP system. This review provides an overview of the cre-loxP system and its various permutations. It addresses the limitations of cre-loxP technology and related considerations for experimental design, and it discusses alternative strategies for site-specific genetic recombination and integration. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  9. 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

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

    PubMed

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

    2004-06-17

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

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

    DTIC Science & Technology

    2007-11-02

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

  12. Strategic Plan for Higher Education, 2008-2017. Executive Summary

    ERIC Educational Resources Information Center

    Fingerhut, Eric D.

    2008-01-01

    If the State of Ohio is to grow and prosper, it must raise the education level of its population. The goal of this 10-year strategic plan is to raise the educational attainment of the state each year, and to close the gap between Ohio and competitor states and nations. To accomplish this goal stakeholders must do three things: (1) Graduate more…

  13. Strategic Plan for Higher Education, 2008-2017

    ERIC Educational Resources Information Center

    Fingerhut, Eric D.

    2008-01-01

    If the State of Ohio is to grow and prosper, it must raise the education level of its population. The goal of this 10-year strategic plan is to raise the educational attainment of the state each year, and to close the gap between Ohio and competitor states and nations. To accomplish this goal, stakeholders must do three things: (1) Graduate more…

  14. 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.

  15. 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

  16. 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.

  17. 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.

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

    PubMed Central

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

    2012-01-01

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

  19. CGRPα within the Trpv1-Cre population contributes to visceral nociception.

    PubMed

    Spencer, Nick J; Magnúsdóttir, Elin I; Jakobsson, Jon; Kestell, Garreth; Nan Chen, Bao; Morris, David; Brookes, Simon J H; Lagerstrom, Malin C

    2017-09-28

    The role of Calcitonin Gene Related Peptide (CGRP) in visceral and somatic nociception is incompletely understood. CGRPα is highly expressed in sensory neurons of dorsal root ganglia (DRG) and particularly in neurons that also express the transient receptor potential cation channel subfamily V member 1 (Trpv1). Therefore, we investigated changes in visceral and somatic nociception following deletion of CGRPα from the Trpv1-Cre population using the Cre/lox system. In control mice, acetic acid injection (0.6%,i.p.) caused significant immobility (time stationary) - an established indicator of visceral pain. In CGRPα-mCherry(lx/lx);Trpv1-Cre mice the duration of immobility was significantly less than controls and the distance CGRPα-mCherry(lx/lx);Trpv1-Cre mice travelled over 20 minutes following acetic acid was significantly greater than controls. However, following acetic acid injection, there was no difference between genotypes in the writhing reflex, number of abdominal licks, or forepaw wipes of the cheek. CGRPα-mCherry(lx/lx);Trpv1-Cre mice developed more pronounced inflammation-induced heat hypersensitivity above baseline values, compared to controls. However, analyses of noxious acute heat or cold transmission revealed no difference between genotypes. Also, odour avoidance test, odour preference test and buried food test for olfaction revealed no differences between genotypes. Our findings suggest that CGRPα-mediated transmission within the Trpv1-Cre population plays a significant role in visceral nociceptive pathways underlying voluntary movement. Monitoring changes in movement over time is a sensitive parameter to identify differences in visceral nociception, compared to writhing reflexes, abdominal licks, or forepaw wipes of the cheek that were unaffected by deletion of CGRPα- from Trpv1-Cre population and likely utilize different mechanisms. Copyright © 2017, American Journal of Physiology-Gastrointestinal and Liver Physiology.

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

    PubMed Central

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

    2015-01-01

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

  1. 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)

  2. 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)

  3. ELT and Consciousness-Raising

    ERIC Educational Resources Information Center

    Al-Jardani, Khalid Salim Saif

    2012-01-01

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

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

    NASA Astrophysics Data System (ADS)

    Faherty, Jacqueline K.; Holford, M.

    2014-01-01

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

  5. Reliable Genetic Labeling of Adult-Born Dentate Granule Cells Using Ascl1 CreERT2 and Glast CreERT2 Murine Lines.

    PubMed

    Yang, Sung M; Alvarez, Diego D; Schinder, Alejandro F

    2015-11-18

    Newly generated dentate granule cells (GCs) are relevant for input discrimination in the adult hippocampus. Yet, their precise contribution to information processing remains unclear. To address this question, it is essential to develop approaches to precisely label entire cohorts of adult-born GCs. In this work, we used genetically modified mice to allow conditional expression of tdTomato (Tom) in adult-born GCs and characterized their development and functional integration. Ascl1(CreERT2);CAG(floxStopTom) and Glast(CreERT2);CAG(floxStopTom) mice resulted in indelible expression of Tom in adult neural stem cells and their lineage upon tamoxifen induction. Whole-cell recordings were performed to measure intrinsic excitability, firing behavior, and afferent excitatory connectivity. Developing GCs were also staged by the expression of early and late neuronal markers. The slow development of adult-born GCs characterized here is consistent with previous reports using retroviral approaches that have revealed that a mature phenotype is typically achieved after 6-8 weeks. Our findings demonstrate that Ascl1(CreERT2) and Glast(CreERT2) mouse lines enable simple and reliable labeling of adult-born GC lineages within restricted time windows. Therefore, these mice greatly facilitate tagging new neurons and manipulating their activity, required for understanding adult neurogenesis in the context of network remodeling, learning, and behavior. Our study shows that Ascl1(CreERT2) and Glast(CreERT2) mice lines can be used to label large cohorts of adult-born dentate granule cells with excellent time resolution. Neurons labeled in this manner display developmental and functional profiles that are in full agreement with previous findings using thymidine analogs and retroviral labeling, thus providing an alternative approach to tackle fundamental questions on circuit remodeling. Because of the massive neuronal targeting and the simplicity of this method, genetic labeling will

  6. 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.

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

    PubMed Central

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

    2010-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  9. Podophyllotoxin induces CREB phosphorylation and CRE-driven gene expression via PKA but not MAPKs.

    PubMed

    Chen, Ya Qiong; Xie, Xin

    2010-01-01

    CRE-driven luciferase reporter is commonly used in drug screening systems involving G protein-coupled receptors (GPCRs). In a screen campaign designed to search for melanocortin-4 receptor (MC4R) agonists, podophyllotoxin, a microtubules disruptor, was found to induce cAMP-responsive element (CRE)-driven reporter expression. MC4R was not involved because podophyllotoxin induced CREB activation and CRE-driven transcription in cells not expressing MC4R. Previous studies indicated that intracellular calcium, PKA, and MAPKs are involved in CREB phosphorylation and activation. Our studies revealed that podophyllotoxin did not affect intracellular calcium level and the phosphorylation state of p38. Podophyllotoxin induced JNK and ERK activation, but blockade of JNK and ERK activation with specific inhibitors had no effect on podophyllotoxin-induced CREB activation and CRE-regulated gene expression. Further experiments revealed that H89, a specific inhibitor of PKA, significantly inhibited podophyllotoxin-induced CREB activation. Podophyllotoxin itself did not alter intracellular cAMP level. Taken together, podophyllotoxin induces CREB activation and CRE-driven gene expression via PKA activation by a cAMP-independent mechanism.

  10. 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.

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

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

    2011-01-01

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

  13. Moving Beyond Strategic Planning to Strategic Thinking.

    ERIC Educational Resources Information Center

    Wolverton, Mimi; Gmelch, Walter

    1999-01-01

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

  14. Plasma is a strategic resource.

    PubMed

    Strengers, Paul F W; Klein, Harvey G

    2016-12-01

    Plasma-derived medicinal products (PDMPs) such as immunoglobulins and clotting factors are listed by the World Health Organization as essential medicines. These and other PDMPs are crucial for the prophylaxis and treatment of patients with bleeding disorders, immune deficiencies, autoimmune and inflammatory diseases, and a variety of congenital deficiency disorders. While changes in clinical practice in developed countries have reduced the need for red blood cell transfusions thereby significantly reducing the collection volumes of whole blood and recovered plasma suitable for fractionation, the need for PDMPs worldwide continues to increase. The majority of plasma supplies for the manufacture of PDMPs is met by the US commercial plasma industry. However, geographic imbalance in the collection of plasma raises concerns that local disruptions of plasma supplies could result in regional and global shortages of essential PDMPs. Plasma, which fits the definition of a strategic resource, that is, "an economically important raw material which is subject to a higher risk of supply interruption," should be considered a strategic resource comparable to energy and drinking water. Plasma collections should be increased outside the United States, including in low- and middle-income countries. The need for capacity building in these countries is an essential part to strengthen quality plasma collection. This will require changes in national and regional policies. We advocate the need for the restoration of an equitable balance of the international plasma supply to reduce the risk of supply shortages worldwide. Strategic independence of plasma should be endorsed on a global level. © 2016 AABB.

  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. Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro.

    PubMed Central

    Aoki, K.; Barker, C.; Danthinne, X.; Imperiale, M. J.; Nabel, G. J.

    1999-01-01

    BACKGROUND: Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system. MATERIALS AND METHODS: Full-length recombinant adenovirus DNA was generated in vitro by Cre-mediated recombination between loxP sites in a linearized shuttle plasmid containing a transgene and adenovirus genomic DNA. RESULTS: After transfection of Cre-treated DNA into 293 cells, replication-defective viral vectors were rapidly obtained without detectable wild-type virus. CONCLUSION: This system facilitates the development of recombinant adenoviral vectors for basic and clinical research. Images Fig. 1 Fig. 2 Fig. 4 PMID:10448644

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

    PubMed

    Ohtsubo, Kazuaki; Marth, Jamey D

    2007-07-01

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

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

  19. 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.

  20. Neurons exhibit Lyz2 promoter activity in vivo: Implications for using LysM-Cre mice in myeloid cell research.

    PubMed

    Orthgiess, Johannes; Gericke, Martin; Immig, Kerstin; Schulz, Angela; Hirrlinger, Johannes; Bechmann, Ingo; Eilers, Jens

    2016-06-01

    To characterize LysM-Cre mediated gene targeting in mice, we crossed LysM-Cre mice to two independent reporter-mouse lines (tdTomato or YFP). Surprisingly, we found that more than 90% of cells with LysM-Cre mediated recombination in the brain were neurons, rather than myeloid cells, such as microglia. Hence, by using the LysM-Cre mouse line for conditional knockout approaches, a significant neuronal recombination needs to be considered. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. 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.

  2. Overlapping ETS and CRE Motifs (G/CCGGAAGTGACGTCA) Preferentially Bound by GABPα and CREB Proteins

    PubMed Central

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

    2012-01-01

    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 (X4-N1-30-X4) to identify pairs of TFBS that localize in proximal promoters at a precise distance. These include two overlapping TFBS: the ETS⇔ETS motif (C/GCCGGAAGCGGAA) and the ETS⇔CRE motif (C/GCGGAAGTGACGTCAC). 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

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

    PubMed Central

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

    2016-01-01

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

  4. 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.

  5. Learning to think strategically.

    PubMed

    1994-01-01

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

  6. 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.

  7. Raising HDL cholesterol in women.

    PubMed

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

    2010-08-09

    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.

  8. Strategic Defense Initiative - strategic implications. Study project

    SciTech Connect

    Russell, T.L.

    1988-03-31

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

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

    PubMed

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

    2013-04-01

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

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

    PubMed Central

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

    1994-01-01

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

  11. Induced Pluripotent Stem Cells Generated from P0-Cre;Z/EG Transgenic Mice.

    PubMed

    Ogawa, Yasuhiro; Eto, Akira; Miyake, Chisato; Tsuchida, Nana; Miyake, Haruka; Takaku, Yasuhiro; Hagiwara, Hiroaki; Oishi, Kazuhiko

    2015-01-01

    Neural crest (NC) cells are a migratory, multipotent cell population that arises at the neural plate border, and migrate from the dorsal neural tube to their target tissues, where they differentiate into various cell types. Abnormal development of NC cells can result in severe congenital birth defects. Because only a limited number of cells can be obtained from an embryo, mechanistic studies are difficult to perform with directly isolated NC cells. Protein zero (P0) is expressed by migrating NC cells during the early embryonic period. In the P0-Cre;Z/EG transgenic mouse, transient activation of the P0 promoter induces Cre-mediated recombination, indelibly tagging NC-derived cells with enhanced green fluorescent protein (EGFP). Induced pluripotent stem cell (iPSC) technology offers new opportunities for both mechanistic studies and development of stem cell-based therapies. Here, we report the generation of iPSCs from the P0-Cre;Z/EG mouse. P0-Cre;Z/EG mouse-derived iPSCs (P/G-iPSCs) exhibited pluripotent stem cell properties. In lineage-directed differentiation studies, P/G-iPSCs were efficiently differentiated along the neural lineage while expressing EGFP. These results suggest that P/G-iPSCs are useful to study NC development and NC-associated diseases.

  12. STIM1fl/fl Ksp-Cre Mouse has Impaired Renal Water Balance.

    PubMed

    Cebotaru, Liudmila; Cebotaru, Valeriu; Wang, Hua; Arend, Lois J; Guggino, William B

    2016-01-01

    STIM1 is as an essential component in store operated Ca2+ entry. However give the paucity of information on the role of STIM1 in kidney, the aim was to study the function of STIM1 in the medulla of the kidney. we crossed a Ksp-cre mouse with another mouse containing two loxP sites flanking Exon 6 of STIM1. The Ksp-cre mouse is based upon the Ksp-cadherin gene promoter which expresses cre recombinase in developing nephrons, collecting ducts (SD) and thick ascending limbs (TAL) of the loop of Henle. The offspring of these mice are viable without gross morphological changes, however, we noticed that the STIM1 Ksp-cre knockout mice produced more urine compared to control. To examine this more carefully, we fed mice low (LP) and high protein (HP) diets respectively. When mice were fed HP diet STIM1 ko mice had significantly increased urinary volume and lower specific gravity compared to wt mice. In STIM1 ko mice fed HP diet urine creatinine and urea were significantly lower compared to wt mice fed HP diet, however the fractional excretion was the same. These data support the idea that STIM1 ko mice have impaired urinary concentrating ability when challenged with HP diet is most likely caused by impaired Ca2+-dependent signal transduction through the vasopressin receptor cascade. © 2016 The Author(s) Published by S. Karger AG, Basel.

  13. 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. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    PubMed Central

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

    2016-01-01

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

  15. 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

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

    ERIC Educational Resources Information Center

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

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

  17. Disruption of Trichoderma reesei cre2, encoding an ubiquitin C-terminal hydrolase, results in increased cellulase activity

    PubMed Central

    2011-01-01

    Background The filamentous fungus Trichoderma reesei (Hypocrea jecorina) is an important source of cellulases for use in the textile and alternative fuel industries. To fully understand the regulation of cellulase production in T. reesei, the role of a gene known to be involved in carbon regulation in Aspergillus nidulans, but unstudied in T. reesei, was investigated. Results The T. reesei orthologue of the A. nidulans creB gene, designated cre2, was identified and shown to be functional through heterologous complementation of a creB mutation in A. nidulans. A T. reesei strain was constructed using gene disruption techniques that contained a disrupted cre2 gene. This strain, JKTR2-6, exhibited phenotypes similar to the A. nidulans creB mutant strain both in carbon catabolite repressing, and in carbon catabolite derepressing conditions. Importantly, the disruption also led to elevated cellulase levels. Conclusions These results demonstrate that cre2 is involved in cellulase expression. Since the disruption of cre2 increases the amount of cellulase activity, without severe morphological affects, targeting creB orthologues for disruption in other industrially useful filamentous fungi, such as Aspergillus oryzae, Trichoderma harzianum or Aspergillus niger may also lead to elevated hydrolytic enzyme activity in these species. PMID:22070776

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

    PubMed

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

    2011-07-01

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

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed

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

    2017-01-01

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

  1. Raising the Bar for Teaching

    ERIC Educational Resources Information Center

    Mehta, Jal; Doctor, Joe

    2013-01-01

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

  2. 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…

  3. 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…

  4. 5-Aminolaevulinate synthase gene promoter contains two cAMP-response element (CRE)-like sites that confer positive and negative responsiveness to CRE-binding protein (CREB).

    PubMed Central

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

    2001-01-01

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

  5. 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.

  6. 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.

  7. 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.

  8. Empirical Questionnaire Methods for Fund-Raising Campaign Preparedness in Extension

    ERIC Educational Resources Information Center

    Comley Adams, Catherine; Butler, Douglass A.

    2017-01-01

    Amid waning public financial support for Extension program offerings, highly strategic and professional fund-raising practices are necessary for gaining momentum among private philanthropists and closing the fiscal gap. University of Missouri Extension conducted a precampaign survey that invited feedback from stakeholders to inform Extension…

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

    PubMed

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

    2014-01-01

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

  10. 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.

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

    PubMed Central

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

    2015-01-01

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

  12. 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.

  13. Keeping the Strategic Flame

    DTIC Science & Technology

    1997-01-01

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

  14. 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.

  15. 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…

  16. 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…

  17. 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.

  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. 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…

  1. Developing Strategic Readers

    ERIC Educational Resources Information Center

    Jones, Jennifer; Leahy, Susie

    2006-01-01

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

  2. THE DMP1-SOST TRANSGENE INTERACTS WITH AND DOWNREGULATES THE DMP1-CRE TRANSGENE AND THE ROSANOTCH ALLELE

    PubMed Central

    Zanotti, Stefano; Canalis, Ernesto

    2016-01-01

    Activation of Notch1 in osteocytes of RosaNotch 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, RosaNotch 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 RosaNotch allele (Dmp1-Cre;RosaNotch) exhibited osteopetrosis. The phenotype of Notch activation in osteocytes was prevented in Dmp1-Cre;RosaNotch 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;RosaNotch mice or from RosaNotch 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;RosaNotch 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

  3. 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.

  4. Generation and characterization of transgenic mice expressing tamoxifen-inducible cre-fusion protein specifically in mouse liver.

    PubMed

    Zhu, Huan-Zhang; Chen, Jian-Quan; Cheng, Guo-Xiang; Xue, Jing-Lun

    2003-08-01

    To establish transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase specifically in the liver and to provide an efficient animal model for studying gene function in the liver and creating various mouse models mimicking human diseases. Alb-Cre-ERt transgenic mice were produced by microinjecting the construct with Cre-ERt fusion gene of DNA fragments into fertilized eggs derived from inbred C57BL/6 strain. Transgenic mice were identified by using PCR and Southern blotting. Expression of Cre-ERt fusion gene was analyzed in the liver, kidney, brain and lung from F1 generation transgenic mice at 8 weeks of age by reverse transcription (RT)-PCR. Four hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant Alb-Cre-ERt DNA fragments, and 312 survival eggs injected were transferred to the oviducts of 12 pseudopregnant recipient mice, 6 of 12 recipient mice became pregnant and gave birth to 44 offsprings. Of the 44 offsprings, two males and one female carried the hybrid Cre-ERt fusion gene. Three mice were determined as founders, and were back crossed to set up F1 generations with other inbred C57BL/6 mice. Transmission of Cre-ERt fusion gene in F1 offspring followed Mendelian rules. The expression of Cre-ERt mRNA was detected only in the liver of F1 offspring from two of three founder mice. Transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase under control of the liver-specific promoter are preliminary established.

  5. 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

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

    PubMed

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

    2005-05-01

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

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

    PubMed

    Bordonaro, Michael

    2009-01-01

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

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

    PubMed Central

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

    2008-01-01

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

  9. 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…

  10. 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

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

    PubMed Central

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

    2015-01-01

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

  12. 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…

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

    DTIC Science & Technology

    2002-05-01

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

  14. 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.

  15. Generating mouse models of degenerative diseases using Cre/lox-mediated in vivo mosaic cell ablation

    PubMed Central

    Fujioka, Masato; Tokano, Hisashi; Fujioka, Keiko Shiina; Okano, Hideyuki; Edge, Albert S.B.

    2011-01-01

    Most degenerative diseases begin with a gradual loss of specific cell types before reaching a threshold for symptomatic onset. However, the endogenous regenerative capacities of different tissues are difficult to study, because of the limitations of models for early stages of cell loss. Therefore, we generated a transgenic mouse line (Mos-iCsp3) in which a lox-mismatched Cre/lox cassette can be activated to produce a drug-regulated dimerizable caspase-3. Tissue-restricted Cre expression yielded stochastic Casp3 expression, randomly ablating a subset of specific cell types in a defined domain. The limited and mosaic cell loss led to distinct responses in 3 different tissues targeted using respective Cre mice: reversible, impaired glucose tolerance with normoglycemia in pancreatic β cells; wound healing and irreversible hair loss in the skin; and permanent moderate deafness due to the loss of auditory hair cells in the inner ear. These mice will be important for assessing the repair capacities of tissues and the potential effectiveness of new regenerative therapies. PMID:21576819

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

    PubMed

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

    2007-02-01

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

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

    PubMed

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

    2013-09-01

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

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed

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

    2017-03-01

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

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

    PubMed

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

    2017-01-01

    RAS signaling is central to many cellular processes and SOS proteins promote RAS activation. To investigate the role of SOS proteins in T cell biology, we crossed Sos1(f/f)Sos2(-/-) mice to CD4-Cre transgenic mice. We previously reported an effect of these mutations on T cell signaling and T cell migration. Unexpectedly, we observed nodules on the joints of greater than 90% of these mutant mice at 5 months of age, especially on the carpal joints. As the mice aged further, some also displayed joint stiffness, hind limb paralysis, and lameness. Histological analysis indicated that the abnormal growth in joints originated from dysplastic chondrocytes. Second harmonic generation imaging of the carpal nodules revealed that nodules were encased by rich collagen fibrous networks. Nodules formed in mice also deficient in RAG2, indicating that conventional T cells, which undergo rearrangement of the T cell antigen receptor, are not required for this phenotype. CD4-Cre expression in a subset of cells, either immune lineage cells (e.g., non-conventional T cells) or non-immune lineage cells (e.g., chondrocytes) likely mediates the dramatic phenotype observed in this study. Disruptions of genes in the RAS signaling pathway are especially likely to cause this phenotype. These results also serve as a cautionary tale to those intending to use CD4-Cre transgenic mice to specifically delete genes in conventional T cells.

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

    NASA Astrophysics Data System (ADS)

    Steentoft, H.; Rabbel, W.

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

  2. 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

  3. 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

  4. Strategic Management or Strategic Planning for Defense?

    DTIC Science & Technology

    1989-02-01

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

  5. 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…

  6. 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…

  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. 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.

  9. Inducible recombination in the cardiac conduction system of minK: CreERT² BAC transgenic mice.

    PubMed

    Arnolds, David E; Moskowitz, Ivan P

    2011-11-01

    Inducible Cre recombination is a powerful technology that allows for spatial and temporal modulation of gene expression in vivo. Diseases of the cardiac conduction system (CCS) pose a significant clinical burden but are not currently well understood at the molecular level. To enable inducible recombination in the murine CCS, we created a minK:CreERT(2) bacterial artificial chromosome (BAC) transgenic mouse line. Cre activity is present after tamoxifen administration in the atrioventricular (AV) node, AV bundle, and bundle branches of adult transgenic mice. We anticipate that by enabling inducible recombination specifically in the AV node, bundle, and bundle branches, minK:CreERT(2) BAC transgenic mice will prove useful in advancing our understanding of CCS disease and function. Copyright © 2011 Wiley-Periodicals, Inc.

  10. Strategic nuclear forces of Britain and France

    SciTech Connect

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

    1984-08-01

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

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed Central

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

    2013-01-01

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

  13. 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.

  14. 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.

  15. 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.

  16. 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. Copyright © 2015 He et al.

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

    PubMed Central

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

    2015-01-01

    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/GCGGAAGT) and CRE (GTGACGTCAC) motifs overlap precisely, producing the ETS↔CRE motif (C/GCGGAAGTGACGTCAC). 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. PMID:26185160

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

    PubMed

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

    2014-11-01

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

  19. Raised Bedrock in Terra Cimmeria

    NASA Image and Video Library

    2017-01-05

    Large impact craters have central hills or mountains, because the tremendous shock waves from the impact first compresses the ground, then causes a rebound when it becomes uncompressed. This effectively raises bedrock that was once deeply buried to the surface. Furthermore, some of the most interesting bedrock on Mars is amongst the oldest and deeply buried. Thus, these crater central uplifts act as windows into ancient Mars, and enable us to peer into a time when certain geologic processes were more active than today. The enhanced colors in this image reflect different bedrock compositions. Some of the large blocks are broken up and jumbled by this impact event or were resampled from previous large impacts. http://photojournal.jpl.nasa.gov/catalog/PIA15149

  20. Engineering Forum Strategic Plan

    EPA Pesticide Factsheets

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

  1. Information and strategic voting.

    PubMed

    Tyszler, Marcelo; Schram, Arthur

    2016-01-01

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

  2. The IAU Strategic Plan

    NASA Astrophysics Data System (ADS)

    Miley, George

    2016-10-01

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

  3. 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.

  4. USACE Navigation Strategic Vision

    DTIC Science & Technology

    2012-12-01

    Initiatives ( SNI ) within these Objectives. Figure 2 illustrates the relationship of SNI to the focus areas and guiding documents, the USACE Campaign...appreciation of the value provided by the MTS and proactive participation with the USACE navigation program. Figure 2. Strategic Navigation Initiatives ( SNI ...Campaign Plan Goal 5, Objective 4a, the CW Strategic Plan Goal 5, and NSV Objective 4 and two supporting SNIs .   7 Objective: Create an inventory of all

  5. IT strategic planning in hospitals: from theory to practice.

    PubMed

    Jaana, Mirou; Teitelbaum, Mari; Roffey, Tyson

    2014-07-01

    To date, IT strategic planning has been mostly theory-based with limited information on "best practices" in this area. This study presents the process and outcomes of IT strategic planning undertaken at a pediatric hospital (PH) in Canada. A five-stage sequential and incremental process was adopted. Various tools / approaches were used including review of existing documentation, internal survey (n = 111), fifteen interviews, and twelve workshops. IT strategic planning was informed by 230 individuals (12 percent of hospital community) and revealed consistency in the themes and concerns raised by participants (e.g., slow IT projects delivery rate, lack of understanding of IT priorities, strained communication with IT staff). Mobile and remote access to patients' information, and an integrated EMR were identified as top priorities. The methodology and used approach revealed effective, improved internal relationships, and ensured commitment to the final IT strategic plan. Several lessons were learned including: maintaining a dynamic approach capable of adapting to the fast technology evolution; involving stakeholders and ensuring continuous communication; using effective research tools to support strategic planning; and grounding the process and final product in existing models. This study contributes to the development of "best practices" in IT strategic planning, and illustrates "how" to apply the theoretical principles in this area. This is especially important as IT leaders are encouraged to integrate evidence-based management into their decision making and practices. The methodology and lessons learned may inform practitioners in other hospitals planning to engage in IT strategic planning in the future.

  6. Strategic Stability Through the Strategic Defense Initiative

    DTIC Science & Technology

    1989-03-09

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

  7. Genetically Targeted All-Optical Electrophysiology with a Transgenic Cre-Dependent Optopatch Mouse

    PubMed Central

    Lou, Shan; Adam, Yoav; Weinstein, Eli N.; Williams, Erika; Williams, Katherine; Parot, Vicente; Kavokine, Nikita; Liberles, Stephen; Madisen, Linda; Zeng, Hongkui

    2016-01-01

    Recent advances in optogenetics have enabled simultaneous optical perturbation and optical readout of membrane potential in diverse cell types. Here, we develop and characterize a Cre-dependent transgenic Optopatch2 mouse line that we call Floxopatch. The animals expressed a blue-shifted channelrhodopsin, CheRiff, and a near infrared Archaerhodopsin-derived voltage indicator, QuasAr2, via targeted knock-in at the rosa26 locus. In Optopatch-expressing animals, we tested for overall health, genetically targeted expression, and function of the optogenetic components. In offspring of Floxopatch mice crossed with a variety of Cre driver lines, we observed spontaneous and optically evoked activity in vitro in acute brain slices and in vivo in somatosensory ganglia. Cell-type-specific expression allowed classification and characterization of neuronal subtypes based on their firing patterns. The Floxopatch mouse line is a useful tool for fast and sensitive characterization of neural activity in genetically specified cell types in intact tissue. SIGNIFICANCE STATEMENT Optical recordings of neural activity offer the promise of rapid and spatially resolved mapping of neural function. Calcium imaging has been widely applied in this mode, but is insensitive to the details of action potential waveforms and subthreshold events. Simultaneous optical perturbation and optical readout of single-cell electrical activity (“Optopatch”) has been demonstrated in cultured neurons and in organotypic brain slices, but not in acute brain slices or in vivo. Here, we describe a transgenic mouse in which expression of Optopatch constructs is controlled by the Cre-recombinase enzyme. This animal enables fast and robust optical measurements of single-cell electrical excitability in acute brain slices and in somatosensory ganglia in vivo, opening the door to rapid optical mapping of neuronal excitability. PMID:27798186

  8. Passive leg raising during pregnancy.

    PubMed

    Marques, Nicole Ribeiro; Martinello, Caroline; Kramer, George C; Costantine, Maged M; Vadhera, Rakesh B; Saade, George R; Hankins, Gary D; Pacheco, Luis D

    2015-03-01

    To determine if passive leg raising (PLR) significantly increases cardiac output in a cohort of healthy pregnant women during the third trimester. Using a noninvasive monitor, baseline hemodynamic measurements for arterial blood pressure, systolic and diastolic blood pressure, heart rate, cardiac output, cardiac index, stroke volume, and systemic vascular resistances were obtained with patients in the semirecumbent position. Measurements were repeated after a 3-minute PLR maneuver in supine, right lateral decubitus, and left lateral decubitus positions. After 10 minutes of bed rest, the cohort's mean baseline heart rate was 80 ± 12 beats/minute. Baseline stroke volume was 98 ± 14 mL, mean cardiac output was 7.8 ± 1.2 L/min, and mean cardiac index was 4.32 ± 0.63 L/min. The baseline systemic vascular resistance value was 893 ± 160 dynes/sec/cm(5). Baseline mean arterial blood pressure was 84 ± 11 mm Hg. Following a PLR maneuver in the supine position, heart rate decreased significantly. No difference was noted in other measurements. Findings were similar with PLR in the left lateral decubitus. PLR in the right lateral decubitus resulted in significantly decreased heart rate, cardiac output, and cardiac index. PLR did not result in cardiac output recruitment in a cohort of healthy pregnant women during the third trimester. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  9. An evaluation of GO annotation retrieval for BioCreAtIvE and GOA.

    PubMed

    Camon, Evelyn B; Barrell, Daniel G; Dimmer, Emily C; Lee, Vivian; Magrane, Michele; Maslen, John; Binns, David; Apweiler, Rolf

    2005-01-01

    The Gene Ontology Annotation (GOA) database http://www.ebi.ac.uk/GOA aims to provide high-quality supplementary GO annotation to proteins in the UniProt Knowledgebase. Like many other biological databases, GOA gathers much of its content from the careful manual curation of literature. However, as both the volume of literature and of proteins requiring characterization increases, the manual processing capability can become overloaded. Consequently, semi-automated aids are often employed to expedite the curation process. Traditionally, electronic techniques in GOA depend largely on exploiting the knowledge in existing resources such as InterPro. However, in recent years, text mining has been hailed as a potentially useful tool to aid the curation process. To encourage the development of such tools, the GOA team at EBI agreed to take part in the functional annotation task of the BioCreAtIvE (Critical Assessment of Information Extraction systems in Biology) challenge. BioCreAtIvE task 2 was an experiment to test if automatically derived classification using information retrieval and extraction could assist expert biologists in the annotation of the GO vocabulary to the proteins in the UniProt Knowledgebase. GOA provided the training corpus of over 9000 manual GO annotations extracted from the literature. For the test set, we provided a corpus of 200 new Journal of Biological Chemistry articles used to annotate 286 human proteins with GO terms. A team of experts manually evaluated the results of 9 participating groups, each of which provided highlighted sentences to support their GO and protein annotation predictions. Here, we give a biological perspective on the evaluation, explain how we annotate GO using literature and offer some suggestions to improve the precision of future text-retrieval and extraction techniques. Finally, we provide the results of the first inter-annotator agreement study for manual GO curation, as well as an assessment of our current electronic

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

    SciTech Connect

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

    1995-12-31

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

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

    PubMed

    Kandel, Eric R

    2012-05-14

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

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

    PubMed Central

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

    2004-01-01

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

  13. 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.

  14. 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.

  15. Moderate and high amounts of tamoxifen in αMHC-MerCreMer mice induce a DNA damage response, leading to heart failure and death

    PubMed Central

    Bersell, Kevin; Choudhury, Sangita; Mollova, Mariya; Polizzotti, Brian D.; Ganapathy, Balakrishnan; Walsh, Stuart; Wadugu, Brian; Arab, Shima; Kühn, Bernhard

    2013-01-01

    SUMMARY Numerous mouse models have utilized Cre-loxP technology to modify gene expression. Adverse effects of Cre recombinase activity have been reported, including in the heart. However, the mechanisms associated with cardiac Cre toxicity are largely unknown. Here, we show that expression of Cre in cardiomyocytes induces a DNA damage response, resulting in cardiomyocyte apoptosis, cardiac fibrosis and cardiac dysfunction. In an effort to increase the recombination efficiency of a widely used tamoxifen-sensitive Cre transgene under control of the α-myosin-heavy-chain promoter (αMHC-MerCreMer), we observed myocardial dysfunction and decreased survival, which were dependent on the dose of tamoxifen injected. After excluding a Cre-independent contribution by tamoxifen, we found that Cre induced myocardial fibrosis, activation of pro-fibrotic genes and cardiomyocyte apoptosis. Examination of the molecular mechanisms showed activation of DNA damage response signaling and p53 stabilization in the absence of loxP sites, suggesting that Cre induced illegitimate DNA breaks. Cardiomyocyte apoptosis was also induced by expressing Cre using adenoviral transduction, indicating that the effect was not dependent on genomic integration of the transgene. Cre-mediated homologous recombination at loxP sites was dose-dependent and had a ceiling effect at ∼80% of cardiomyocytes showing recombination. By titrating the amount of tamoxifen to maximize recombination while minimizing animal lethality, we determined that 30 μg tamoxifen/g body weight/day injected on three consecutive days is the optimal condition for the αMHC-MerCreMer system to induce recombination in the Rosa26-lacZ strain. Our results further highlight the importance of experimental design, including the use of appropriate genetic controls for Cre expression. PMID:23929941

  16. 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,…

  17. 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,…

  18. Targeting of GFP-Cre to the Mouse Cyp11a1 Locus Both Drives Cre Recombinase Expression in Steroidogenic Cells and Permits Generation of Cyp11a1 Knock Out Mice

    PubMed Central

    O'Hara, Laura; York, Jean Philippe; Zhang, Pumin; Smith, Lee B.

    2014-01-01

    To permit conditional gene targeting of floxed alleles in steroidogenic cell-types we have generated a transgenic mouse line that expresses Cre Recombinase under the regulation of the endogenous Cytochrome P450 side chain cleavage enzyme (Cyp11a1) promoter. Mice Carrying the Cyp11a1-GC (GFP-Cre) allele express Cre Recombinase in fetal adrenal and testis, and adrenal cortex, testicular Leydig cells (and a small proportion of Sertoli cells), theca cells of the ovary, and the hindbrain in postnatal life. Circulating testosterone concentration is unchanged in Cyp11+/GC males, suggesting steroidogenesis is unaffected by loss of one allele of Cyp11a1, mice are grossly normal, and Cre Recombinase functions to recombine floxed alleles of both a YFP reporter gene and the Androgen Receptor (AR) in steroidogenic cells of the testis, ovary, adrenal and hindbrain. Additionally, when bred to homozygosity (Cyp11a1GC/GC), knock-in of GFP-Cre to the endogenous Cyp11a1 locus results in a novel mouse model lacking endogenous Cyp11a1 (P450-SCC) function. This unique dual-purpose model has utility both for those wishing to conditionally target genes within steroidogenic cell types and for studies requiring mice lacking endogenous steroid hormone production. PMID:24404170

  19. 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.

  20. 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

  1. Biallelic Dicer1 loss mediated by aP2-Cre drives angiosarcoma.

    PubMed

    Hanna, Jason A; Drummond, Catherine J; Garcia, Matthew R; Go, Jonathan C; Finkelstein, David; Rehg, Jerold E; Hatley, Mark E

    2017-09-15

    Angiosarcoma is an aggressive vascular sarcoma with an extremely poor prognosis. Due to the relative rarity of this disease, its molecular drivers and optimal treatment strategies are obscure. DICER1 is an RNase III endoribonuclease central to microRNA biogenesis, and germline DICER1 mutations result in a cancer predisposition syndrome, associated with an increased risk of many tumor types. Here we show that biallelic Dicer1 deletion with aP2-Cre drives aggressive and metastatic angiosarcoma independent of other genetically engineered oncogenes or tumor suppressor loss. Angiosarcomas in aP2-Cre;Dicer1Flox/- mice histologically and genetically resemble human angiosarcoma. MicroRNA-23 target genes including the oncogenes Ccnd1 as well as Adam19, Plau, and Wsb1 that promote invasiveness and metastasis were enriched in mouse and human angiosarcoma. These studies illustrate that Dicer1 can function as a traditional loss-of-function tumor suppressor gene, and they provide a fully penetrant animal model for the study of angiosarcoma development and metastasis. Copyright ©2017, American Association for Cancer Research.

  2. 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

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

    PubMed

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

    2016-02-01

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

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

    PubMed

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

    2014-11-01

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

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

    PubMed Central

    Niesner, Bradley; Maheshri, Narendra

    2013-01-01

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

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

    PubMed

    Bruns, Mark A; Gibbs, Elizabeth R

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

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

    PubMed Central

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

    2009-01-01

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

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. Cre-mediated germline mosaicism: a method allowing rapid generation of several alleles of a target gene.

    PubMed

    Holzenberger, M; Lenzner, C; Leneuve, P; Zaoui, R; Hamard, G; Vaulont, S; Bouc, Y L

    2000-11-01

    Conditional gene targeting uses the insertion of expression cassettes for the selection of targeted embryonic stem cells. The presence of these cassettes in the final targeted chromosomal locus may affect the normal expression of the targeted gene and produce interesting knock down phenotypes. We show here that the selection cassette may then be selectively removed in vivo, using three appropriately positioned loxP sites in the targeted gene and the transgenic mouse EIIaCre. This strategy was applied to two different target genes and we demonstrated that it is reliable and reproducible. First, we generated double transgenic EIIaCre/loxP mice (F1) that showed variable degrees of mosaicism for partially CRE-recombined floxed alleles. Efficiency of EIIaCre at creating mosaicism was dependent on the target gene and on parental transmission of the transgene. The segregation of partially recombined alleles and EIIaCre transgene was obtained in the next generation using mosaic F1 males. Mosaic females were unsuitable for this purpose because they systematically generated complete excisions during oogenesis. Our strategy is applicable to other approaches based on three loxP sites. As this procedure allows generation of knock down (presence of neo), knockout (total exision of the loxP-flanked sequences) and floxed substrains (excision of the selection cassette) from a single, targeted germline mutation and in a single experiment, its use may become more widespread in conditional mutagenesis.

  15. 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. © 2015 The Authors. European Journal of Immunology published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. 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

  17. 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.

  18. Thinking strategically about capitation.

    PubMed

    Boland, P

    1997-05-01

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

  19. Strategic Analysis of Terrorism

    NASA Astrophysics Data System (ADS)

    Arce, Daniel G.; Sandler, Todd

    Two areas that are increasingly studied in the game-theoretic literature on terrorism and counterterrorism are collective action and asymmetric information. One contribution of this chapter is a survey and extension of continuous policy models with differentiable payoff functions. In this way, policies can be characterized as strategic substitutes (e. g., proactive measures), or strategic complements (e. g., defensive measures). Mixed substitute-complement models are also introduced. We show that the efficiency of counterterror policy depends upon (i) the strategic substitutes-complements characterization, and (ii) who initiates the action. Surprisingly, in mixed-models the dichotomy between individual and collective action may disappear. A second contribution is the consideration of a signaling model where indiscriminant spectacular terrorist attacks may erode terrorists’ support among its constituency, and proactive government responses can create a backlash effect in favor of terrorists. A novel equilibrium of this model reflects the well-documented ineffectiveness of terrorism in achieving its stated goals.

  20. 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

  1. 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…

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

    ERIC Educational Resources Information Center

    Dickmeyer, Nathan

    2004-01-01

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

  3. Strategic avionics technology planning

    NASA Astrophysics Data System (ADS)

    Cox, Kenneth J.; Brown, Don C.

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

  4. On operator strategic behavior

    NASA Technical Reports Server (NTRS)

    Hancock, P. A.

    1991-01-01

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

  5. Strategic avionics technology planning

    NASA Technical Reports Server (NTRS)

    Cox, Kenneth J.; Brown, Don C.

    1991-01-01

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

  6. 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.

  7. Microgravity strategic planning exercise

    NASA Technical Reports Server (NTRS)

    Halpern, Richard; Downey, Jim; Harvey, Harold

    1991-01-01

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

  8. 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.

  9. Strategic Management in the Community College.

    ERIC Educational Resources Information Center

    Myran, Gunder A.

    1983-01-01

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

  10. R/L, a double reporter mouse line that expresses luciferase gene upon Cre-mediated excision, followed by inactivation of mRFP expression.

    PubMed

    Jia, Junshuang; Lin, Xiaolin; Lin, Xia; Lin, Taoyan; Chen, Bangzhu; Hao, Weichao; Cheng, Yushuang; Liu, Yu; Dian, Meijuan; Yao, Kaitai; Xiao, Dong; Gu, Weiwang

    2016-10-01

    The Cre/loxP system has become an important tool for the conditional gene knockout and conditional gene expression in genetically engineered mice. The applications of this system depend on transgenic reporter mouse lines that provide Cre recombinase activity with a defined cell type-, tissue-, or developmental stage-specificity. To develop a sensitive assay for monitoring Cre-mediated DNA excisions in mice, we generated Cre-mediated excision reporter mice, designated R/L mice (R/L: mRFP(monomeric red fluorescent protein)/luciferase), express mRFP throughout embryonic development and adult stages, while Cre-mediated excision deletes a loxP-flanked mRFP reporter gene and STOP sequence, thereby activating the expression of the second reporter gene luciferase, as assayed by in vivo and ex vivo bioluminescence imaging. After germ line deletion of the floxed mRFP and STOP sequence in R/L mice by EIIa-Cre mice, the resulting luciferase transgenic mice in which the loxP-mRFP-STOP-loxP cassette is excised from all cells express luciferase in all tissues and organs examined. The expression of luciferase transgene was activated in liver of RL/Alb-Cre double transgenic mice and in brain of RL/Nestin-Cre double transgenic mice when R/L reporter mice were mated with Alb-Cre mice and Nestin-Cre mice, respectively. Our findings reveal that the double reporter R/L mouse line is able to indicate the occurrence of Cre-mediated excision from early embryonic to adult lineages. Taken together, these findings demonstrate that the R/L mice serve as a sensitive reporter for Cre-mediated DNA excision both in living animals and in organs, tissues, and cells following necropsy.

  11. A Handbook for Strategic Planning

    DTIC Science & Technology

    1994-01-01

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

  12. Strategic plan for infrastructure optimization

    SciTech Connect

    Donley, C.D.

    1998-05-27

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

  13. Establishment of a Cre/loxP recombination system for N-terminal epitope tagging of genes in Tetrahymena.

    PubMed

    Busch, Clara Jana-Lui; Vogt, Alexander; Mochizuki, Kazufumi

    2010-07-13

    Epitope tagging is a powerful strategy to study the function of proteins. Although tools for C-terminal protein tagging in the ciliated protozoan Tetrahymena thermophila have been developed, N-terminal protein tagging in this organism is still technically demanding. In this study, we have established a Cre/loxP recombination system in Tetrahymena and have applied this system for the N-terminal epitope tagging of Tetrahymena genes. Cre can be expressed in Tetrahymena and localizes to the macronucleus where it induces precise recombination at two loxP sequences in direct orientation in the Tetrahymena macronuclear chromosome. This Cre/loxP recombination can be used to remove a loxP-flanked drug-resistance marker from an N-terminal tagging construct after it is integrated into the macronucleus. The system established in this study allows us to express an N-terminal epitope tagged gene from its own endogenous promoter in Tetrahymena.

  14. 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…

  15. 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…

  16. 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…

  17. 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…

  18. 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…

  19. 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…

  20. 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…

  1. Transient Cnp expression by early progenitors causes Cre-Lox-based reporter lines to map profoundly different fates.

    PubMed

    Tognatta, Reshmi; Sun, Wenjing; Goebbels, Sandra; Nave, Klaus-Armin; Nishiyama, Akiko; Schoch, Susanne; Dimou, Leda; Dietrich, Dirk

    2017-02-01

    NG2 expressing oligodendroglial precursor cells are ubiquitous in the central nervous system and the only cell type cycling throughout life. Previous fate mapping studies have remained inconsistent regarding the question whether NG2 cells are capable of generating certain types of neurons. Here, we use CNP-Cre mice to map the fate of a sub-population of NG2 cells assumed to be close to differentiation. When crossing these mice with the ROSA26/YFP Cre-reporter line we discovered large numbers of reporter-expressing pyramidal neurons in the piriform and dorsal cortex. In contrast, when using Z/EG reporter mice to track the fate of Cnp-expressing NG2 cells only oligodendroglial cells were found reporter positive. Using BrdU-based birth dating protocols and inducible NG2CreER:ROSA26/YFP mice we show that YFP positive neurons are generated from radial glial cells and that these radial glial cells display temporary and low level activity of certain oligodendroglial genes sufficient to recombine the Cre-inducible reporter gene in ROSA26/YFP but not in Z/EG mice. Taken together, we did not obtain evidence for generation of neurons from NG2 cells. Our results suggest that with an appropriate reporter system Cnp activity can be used to define a proliferative subpopulation of NG2 cells committed to generate oligodendrocytes. However, the strikingly different results obtained from ROSA26/YFP versus Z/EG mice demonstrate that the choice of Cre-reporter line can be of crucial importance for fate mapping studies and other applications of the Cre-lox technology. GLIA 2017;65:342-359. © 2016 Wiley Periodicals, Inc.

  2. 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)…

  3. 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…

  4. Measuring strategic success.

    PubMed

    Gish, Ryan

    2002-08-01

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

  5. Strategic Education Research Partnership.

    ERIC Educational Resources Information Center

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

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

  6. The Strategic Revolution.

    PubMed

    Gardner, Andy

    2016-09-08

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

  7. 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.

  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 Education Research Partnership.

    ERIC Educational Resources Information Center

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

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

  10. Strategic Defense Initiative program

    SciTech Connect

    Conachan, F.C.

    1991-05-01

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

  11. Strategic Information Systems Planning.

    ERIC Educational Resources Information Center

    Rowley, Jennifer

    1995-01-01

    Strategic Information Systems Planning (SISP) is the process of establishing a program for implementation and use of information systems in ways that will optimize effectiveness of information resources and use them to support the objectives of the organization. Basic steps in SISP methodology are outlined. (JKP)

  12. Strategic Enrollment Management Revolution.

    ERIC Educational Resources Information Center

    Black, Jim, Ed.

    The chapters of this collection describe a transformational period in higher education. The paradigm shift in higher education requires new organizational models, new strategies, new resources, and new tools, with new ways of thinking, especially about strategic enrollment management (SEM). The chapters of part 1, "The Contextual Framework for…

  13. What is strategic management?

    PubMed

    Jasper, Melanie; Crossan, Frank

    2012-10-01

    To discuss the theoretical concept of strategic management and explore its relevance for healthcare organisations and nursing management. Despite being a relatively new approach, the growth of strategic management within organisations has been consistently and increasingly promoted. However, comprehensive definitions are scarce and commonalities of interpretation are limited. This paper presents an exploratory discussion of the construct of strategic management, drawing on the literature and questioning its relevance within health-care organisations. Literature relating to strategic management across a number of fields was accessed, drawing primarily on meta-studies within management literature, to identify key concepts and attempt to present a consistent definition. The concept within health care is explored in relation to nursing management. Inconsistency in definitions and utilisation of key concepts within this management approach results in the term being loosely applied in health-care organisations without recourse to foundational principles and a deep understanding of the approach as a theory as opposed to an applied term. Nurse managers are increasingly asked to adopt the 'next-best-thing' in managerial theories, yet caution needs to be taken in nurses agreeing to use systems that lack an evidence base in terms of both efficacy and relevance of context. © 2012 Blackwell Publishing Ltd.

  14. 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)

  15. 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…

  16. Transformation and Strategic Surprise

    DTIC Science & Technology

    2005-04-01

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

  17. 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…

  18. 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.

  19. 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…

  20. Strategic Sustainability Assessment

    DTIC Science & Technology

    2007-12-01

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

  1. 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.

  2. 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)

  3. 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.

  4. 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…

  5. Strategic Planning Is Back.

    ERIC Educational Resources Information Center

    Galagan, Patricia A.

    1997-01-01

    After an absence of 10 years, strategic planning has returned as a prime management issue. New attitudes, new approaches, and new players have emerged with new practices. A good strategy can make a company more attractive to customers and investors as well as a better place to work. (JOW)

  6. Normal mammary development and function in mice with Ift88 deleted in MMTV- and K14-Cre expressing cells

    PubMed Central

    2014-01-01

    Background Primary cilia (PC) are non-motile microtubule based organelles present on almost every cell type and are known to serve as critical organizing centers for several signaling pathways crucial to embryonic and postnatal development. Alterations in the Hh pathway, the most studied signaling pathway regulated by PC, affect mammary gland development as well as maintenance of the stem and progenitor cell populations. Results We developed mouse models with deletion of PC in mammary luminal epithelial, basal epithelial, and stromal cells for evaluation of the function of PC in mammary development via MMTV-Cre, K14-Cre, and Prx1-Cre mediated deletion, respectively. The activity of Cre was confirmed using ROSA26 reporters. Mammary stem and progenitor cells were enriched through growth as mammospheres. Adenovirus-Cre mediated deletion of Ift88 was used to determine a role for PC in this population of cells. Disruption of Ift88 and PC were confirmed in using PCR and immunofluorescent methods. Prx1-Cre; Ift88Del mice demonstrated defects in terminal end buds during puberty. However, these Ift88Del glands exhibited typical terminal end bud formation as well as normal ductal histology when transplanted into wild type hosts, indicating that the phenotype observed was not intrinsic to the mammary gland. Furthermore, no discernable alterations to mammary development were observed in MMTV-Cre- or K14-Cre; Ift88Del lines. These mice were able to feed and support several litters of pups even though wide spread depletion of PC was confirmed. Cells grown in mammosphere culture were enriched for PC containing cells suggesting PC are preferentially expressed on mammary stem and progenitor cells. Deletion of Ift88 in mammary epithelial cells resulted in a significant reduction in the number of primary mammospheres established; however, there was no effect on outgrowth of secondary mammospheres in PC-depleted cells. Conclusions PC regulate systemic factors that can affect mammary

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

    PubMed

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

    2013-01-01

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

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

    PubMed

    Girlich, Delphine; Poirel, Laurent; Nordmann, Patrice

    2013-02-01

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

  9. Strategic arms limitation

    NASA Astrophysics Data System (ADS)

    Allen Greb, G.; Johnson, Gerald W.

    1983-10-01

    Following World War II, American scientists and politicians proposed in the Baruch plan a radical solution to the problem of nuclear weapons: to eliminate them forever under the auspices of an international nuclear development authority. The Soviets, who as yet did not possess the bomb, rejected this plan. Another approach suggested by Secretary of War Henry Stimson to negotiate directly with the Soviet Union was not accepted by the American leadership. These initial arms limitation failures both reflected and exacerbated the hostile political relationship of the superpowers in the 1950s and 1960s. Since 1969, the more modest focus of the Soviet-American arms control process has been on limiting the numbers and sizes of both defensive and offensive strategic systems. The format for this effort has been the Strategic Arms Limitatins Talks (Salt) and more recently the Strategic Arms Reduction Talks (START). Both sides came to these negotiations convinced that nuclear arsenals had grown so large that some for of mutual restraint was needed. Although the SALT/START process has been slow and ponderous, it has produced several concrete the agreements and collateral benefits. The 1972 ABM Treaty restricts the deployment of ballistic missile defense systems, the 1972 Interim Agreement places a quantitative freeze on each side's land based and sea based strategic launchers, and the as yet unratified 1979 SALT II Treaty sets numerical limits on all offensive strategic systems and sublimits on MIRVed systems. Collateral benefits include improved verification procedures, working definitions and counting rules, and permanent bureaucratic apparatus which enhance stability and increase the chances for achieving additional agreements.

  10. 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.

  11. Cre-mediated reversible immortalization of human renal proximal tubular epithelial cells.

    PubMed

    Kowolik, Claudia M; Liang, Shujian; Yu, Ying; Yee, Jiing-Kuan

    2004-08-05

    Primary human renal proximal tubule epithelial cells (RPTECs) are of limited use for basic research and for clinical applications due to their limited lifespan in culture. Here we used two lentivirus vectors carrying the human telomerase (hTERT) and the SV40T antigen (Tag) flanked by loxP sites to reversibly immortalize RPTECs. Transduced RPTEC clones continued to proliferate while retaining biochemical and functional characteristics of primary cells. The clones exhibited contact-inhibited, anchorage- and growth factor-dependent growth and did not form tumors in nude mice, suggesting that the cells were not transformed. Transient Cre expression in these cells led to efficient proviral deletion, upregulation of some renal specific activities, and decreased growth rates. Ultimately, the cells underwent replicative senescence, indicating intact cell cycle control. Thus, reversible immortalization allows the expansion of human RPTECs, leading to large production of RPTECs that retain most tissue-specific properties.

  12. 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.

  13. 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

  14. Raised intracranial pressure in Apert syndrome.

    PubMed

    Marucci, Damian D; Dunaway, David J; Jones, Barry M; Hayward, Richard D

    2008-10-01

    Raised intracranial pressure is a well-known complication of Apert syndrome. The current policy in the authors' unit is to monitor these patients and only perform surgery when raised intracranial pressure has been diagnosed. The authors present their experience with this protocol, as it allows a more accurate picture of the natural history of raised intracranial pressure in Apert syndrome. The records of 24 patients, aged between 7 and 14 years, with Apert syndrome who had been managed expectantly (i.e., with no routine "automatic" early surgery) were reviewed. Data were collected on the incidence, timing, and management of raised intracranial pressure. Twenty of 24 patients (83 percent) developed raised intracranial pressure. The average age of the first episode was 18 months (range, 1 month to 4 years 5 months). Raised intracranial pressure was managed with surgery in 18 patients, including two patients who underwent shunt procedures for hydrocephalus. Two patients had their raised intracranial pressure treated successfully by correcting coexisting upper airway obstruction alone. Seven of the 20 patients (35 percent) developed a second episode of raised intracranial pressure, on average 3 years 4 months later (range, 1 year 11 months to 5 years 9 months). In Apert syndrome, there is a high incidence of raised intracranial pressure, which can first occur at any age up to 5 years and may recur despite initial successful treatment. Causes of raised intracranial pressure include craniocerebral disproportion, venous hypertension, upper airway obstruction, and hydrocephalus. Careful clinical, ophthalmologic, respiratory, and radiologic monitoring will allow raised intracranial pressure to be diagnosed accurately when it occurs and then treated most appropriately.

  15. Hemizygous Le-Cre Transgenic Mice Have Severe Eye Abnormalities on Some Genetic Backgrounds in the Absence of LoxP Sites

    PubMed Central

    Dorà, Natalie J.; Collinson, J. Martin; Hill, Robert E.; West, John D.

    2014-01-01

    Eye phenotypes were investigated in Le-CreTg/−; Pax6fl/+ mice, which were expected to show tissue-specific reduction of Pax6 in surface ectoderm derivatives. To provide a better comparison with our previous studies of Pax6+/− eye phenotypes, hemizygous Le-CreTg/− and heterozygous Pax6fl/+mice were crossed onto the CBA/Ca genetic background. After the Le-Cre transgene had been backcrossed to CBA/Ca for seven generations, significant eye abnormalities occurred in some hemizygous Le-CreTg/−; Pax6+/+ controls (without a floxed Pax6fl allele) as well as experimental Le-CreTg/−; Pax6fl/+ mice. However, no abnormalities were seen in Le-Cre−/−; Pax6fl/+ or Le-Cre−/−; Pax6+/+ controls (without the Le-Cre transgene). The severity and frequency of the eye abnormalities in Le-CreTg/−; Pax6+/+ control mice diminished after backcrossing Le-CreTg/− mice to the original FVB/N strain for two generations, showing that the effect was reversible. This genetic background effect suggests that the eye abnormalities are a consequence of an interaction between the Le-Cre transgene and alleles of unknown modifier genes present in certain genetic backgrounds. The abnormalities were also ameliorated by introducing additional Pax6 gene copies on a CBA/Ca background, suggesting involvement of Pax6 depletion in Le-CreTg/−; Pax6+/+ mice rather than direct action of Cre recombinase on cryptic pseudo-loxP sites. One possibility is that expression of Cre recombinase from the Pax6-Le regulatory sequences in the Le-Cre transgene depletes cofactors required for endogenous Pax6 gene expression. Our observation that eye abnormalities can occur in hemizygous Le-CreTg/−; Pax6+/+ mice, in the absence of a floxed allele, demonstrates the importance of including all the relevant genetic controls in Cre-loxP experiments. PMID:25272013

  16. Educational Fund Raising and the Law.

    ERIC Educational Resources Information Center

    Bargerstock, Charles T.

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

  17. Asymmetry in Children's Comprehension of Raising

    ERIC Educational Resources Information Center

    Choe, Jinsun; O'Grady, William

    2017-01-01

    This paper investigates English-speaking children's acquisition of raising constructions (e.g. "John seems to Mary to be happy") and fnds an asymmetric effect of NP type on their comprehension: an improvement in performance is observed when a lexical NP is raised across a pronominal experiencer (e.g. "John seems to her to be…

  18. 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…

  19. Adults raised as children in lesbian families.

    PubMed

    Tasker, F; Golombok, S

    1995-04-01

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

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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…

  6. 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…

  7. 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…

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

    PubMed

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

    2017-03-07

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

  9. 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.

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

    PubMed

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

    2013-01-01

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

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

    PubMed Central

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

    2011-01-01

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

  12. Cre-mediated cell ablation contests mast cell contribution in models of antibody- and T cell-mediated autoimmunity.

    PubMed

    Feyerabend, Thorsten B; Weiser, Anne; Tietz, Annette; Stassen, Michael; Harris, Nicola; Kopf, Manfred; Radermacher, Peter; Möller, Peter; Benoist, Christophe; Mathis, Diane; Fehling, Hans Jörg; Rodewald, Hans-Reimer

    2011-11-23

    Immunological functions of mast cells remain poorly understood. Studies in Kit mutant mice suggest key roles for mast cells in certain antibody- and T cell-mediated autoimmune diseases. However, Kit mutations affect multiple cell types of both immune and nonimmune origin. Here, we show that targeted insertion of Cre-recombinase into the mast cell carboxypeptidase A3 locus deleted mast cells in connective and mucosal tissues by a genotoxic Trp53-dependent mechanism. Cre-mediated mast cell eradication (Cre-Master) mice had, with the exception of a lack of mast cells and reduced basophils, a normal immune system. Cre-Master mice were refractory to IgE-mediated anaphylaxis, and this defect was rescued by mast cell reconstitution. This mast cell-deficient strain was fully susceptible to antibody-induced autoimmune arthritis and to experimental autoimmune encephalomyelitis. Differences comparing Kit mutant mast cell deficiency models to selectively mast cell-deficient mice call for a systematic re-evaluation of immunological functions of mast cells beyond allergy. Copyright © 2011 Elsevier Inc. All rights reserved.

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

    PubMed Central

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

    2017-01-01

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

  14. Osterix-Cre Labeled Progenitor Cells Contribute to the Formation and Maintenance of the Bone Marrow Stroma

    PubMed Central

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

    2013-01-01

    We have carried out fate mapping studies using Osterix-EGFPCre and Osterix-CreERt animal models and found Cre reporter expression in many different cell types that make up the bone marrow stroma. Constitutive fate mapping resulted in the labeling of different cellular components located throughout the bone marrow, whereas temporal fate mapping at E14.5 resulted in the labeling of cells within a region of the bone marrow. The identity of cell types marked by constitutive and temporal fate mapping included osteoblasts, adipocytes, vascular smooth muscle, perineural, and stromal cells. Prolonged tracing of embryonic precursors labeled at E14.5dpc revealed the continued existence of their progeny up to 10 months of age, suggesting that fate mapped, labeled embryonic precursors gave rise to long lived bone marrow progenitor cells. To provide further evidence for the marking of bone marrow progenitors, bone marrow cultures derived from Osterix-EGFPCre/Ai9 mice showed that stromal cells retained Cre reporter expression and yielded a FACS sorted population that was able to differentiate into osteoblasts, adipocytes, and chondrocytes in vitro and into osteoblasts, adipocytes, and perivascular stromal cells after transplantation. Collectively, our studies reveal the developmental process by which Osterix-Cre labeled embryonic progenitors give rise to adult bone marrow progenitors which establish and maintain the bone marrow stroma. PMID:23951132

  15. Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation.

    PubMed

    Duncan, Samuel M; Myburgh, Elmarie; Philipon, Cintia; Brown, Elaine; Meissner, Markus; Brewer, James; Mottram, Jeremy C

    2016-06-01

    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. © 2016 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd.

  16. Advanced strategic missile development

    NASA Astrophysics Data System (ADS)

    Strickler, R. L.

    1981-05-01

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

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

    PubMed Central

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

    2017-01-01

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

  18. Strategic Leadership Development Model

    DTIC Science & Technology

    2012-03-19

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

  19. Installation Strategic Planning Guidebook

    DTIC Science & Technology

    2012-05-01

    factors (CSFs) are those areas in which things must go right for the organization to excel. The CSF concept was first introduced by John F. Rockhart in...11. Essential Challenges of Strategic Management by Dr. William B. Rouse, John Wiley & Sons, Inc, Wiley-Interscience Publications, 2001...Strategy Process by H. Mintzberg, J.B.Quinn, and R.M. James , Prentice-Hall, 1988 15. The Way of Strategy by William A. Levinson, ASQC Quality Press

  20. The French Strategic Dilemma.

    DTIC Science & Technology

    1984-03-01

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

  1. Wargaming Strategic Linkage

    DTIC Science & Technology

    2009-01-01

    Low then high 19 The melded game approach 20 Time and the game clock 21 Telescoping time 22 Common time 23 Time jumps 26 Hybrid concepts...naval forces. The introduction of a new maritime strategy in late 2007 triggered renewed interest in the idea of a Global War Game. The new series...this new series, Global 󈧌, conducted in the summer of 2008, took the form of a facilitated workshop focused on strategic-level issues. In future

  2. A National Strategic Narrative

    DTIC Science & Technology

    2011-01-01

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

  3. 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.

  4. Information Technology Strategic Plan

    DTIC Science & Technology

    1998-06-01

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

  5. 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.

  6. Making Strategic Analysis Matter

    DTIC Science & Technology

    2012-01-01

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

  7. Ablation of Foxl1-Cre-labeled hepatic progenitor cells and their descendants impairs recovery of mice from liver injury.

    PubMed

    Shin, Soona; Upadhyay, Naman; Greenbaum, Linda E; Kaestner, Klaus H

    2015-01-01

    Foxl1(+) hepatic progenitor cells (HPCs) differentiate into cholangiocytes and hepatocytes after liver injury. We investigated the requirement for Foxl1(+) HPCs in recovery from liver injury in mice. We developed mice in which we could trace and delete Foxl1-expressing HPCs and their descendants (Foxl1-Cre;Rosa(YFP/iDTR)-inducible diphtheria toxin receptor [iDTR] mice). Foxl1-Cre-negative mice were used as controls. Liver damage was induced in male mice by placing them on choline-deficient, ethionine-supplemented (CDE) diets for 15 days; mice then were placed on normal diets and allowed to recover. Liver damage was induced in female mice by placing them on 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-containing diets, followed by a recovery period. Some mice were given injections of diphtheria toxin during the recovery phase to delete Foxl1-Cre-marked HPCs and their descendants. Livers were collected from all mice and analyzed by immunofluorescence, quantitative reverse-transcription polymerase chain reaction, flow cytometry, and histologic analyses. Foxl1-Cre-marked HPCs were required for the development of cholangiocytes and hepatocytes in livers after CDE diet-induced injury. A smaller percentage of yellow fluorescent protein-positive (YFP(+)) hepatocytes contained markers of oxidative stress, DNA damage, or cell death than YFP-negative hepatocytes, indicating that YFP(+) hepatocytes are newly formed cells. Injection of diphtheria toxin deleted YFP(+) cells from Foxl1-Cre;Rosa(YFP/iDTR) mice and prevented the resolution of hepatic steatosis. In mice recovering from DDC diet-induced injury, most cholangiocytes arose from Foxl1-Cre-marked HPCs. Deletion of YFP(+) cells did not alter levels of markers of liver injury or liver function. Based on studies of Foxl1-Cre;Rosa(YFP/iDTR) mice, Foxl1(+) HPCs and/or their descendants are required for the development of cholangiocytes and hepatocytes in liver after CDE diet-induced injury. Copyright © 2015 AGA Institute

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

    PubMed Central

    Ma, Wenxin; Wang, Shu-Zhen

    2012-01-01

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

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

    PubMed

    Insel, T R; Scolnick, E M

    2006-01-01

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

  10. Soviet strategic defense technology

    SciTech Connect

    Stubbs, E.

    1987-04-01

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

  11. Linking Faculty Raises to College Resources

    ERIC Educational Resources Information Center

    Miller, John

    2006-01-01

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

  12. Modeling distinct osteosarcoma subtypes in vivo using Cre:lox and lineage-restricted transgenic shRNA.

    PubMed

    Mutsaers, Anthony J; Ng, Alvin J M; Baker, Emma K; Russell, Megan R; Chalk, Alistair M; Wall, Meaghan; Liddicoat, Brain J J; Ho, Patricia W M; Slavin, John L; Goradia, Ankita; Martin, T John; Purton, Louise E; Dickins, Ross A; Walkley, Carl R

    2013-07-01

    Osteosarcoma is the most common primary cancer of bone and one that predominantly affects children and adolescents. Osteoblastic osteosarcoma represents the major subtype of this tumor, with approximately equal representation of fibroblastic and chondroblastic subtypes. We and others have previously described murine models of osteosarcoma based on osteoblast-restricted Cre:lox deletion of Trp53 (p53) and Rb1 (Rb), resulting in a phenotype most similar to fibroblastic osteosarcoma in humans. We now report a model of the most prevalent form of human osteosarcoma, the osteoblastic subtype. In contrast to other osteosarcoma models that have used Cre:lox mediated gene deletion, this model was generated through shRNA-based knockdown of p53. As is the case with the human disease the shRNA tumors most frequently present in the long bones and preferentially disseminate to the lungs; feature less consistently modeled using Cre:lox approaches. Our approach allowed direct comparison of the in vivo consequences of targeting the same genetic drivers using two different technologies, Cre:lox and shRNA. This demonstrated that the effects of Cre:lox and shRNA mediated knock-down are qualitatively different, at least in the context of osteosarcoma, and yielded distinct subtypes of osteosarcoma. Through the use of complementary genetic modification strategies we have established a model of the most common clinical subtype of osteosarcoma that was not previously represented and more fully recapitulated the clinical spectrum of this cancer. Copyright © 2013 Elsevier Inc. All rights reserved.

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

    Code of Federal Regulations, 2014 CFR

    2014-10-01

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

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

    Code of Federal Regulations, 2012 CFR

    2012-10-01

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

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

    Code of Federal Regulations, 2011 CFR

    2011-10-01

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

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

    Code of Federal Regulations, 2013 CFR

    2013-10-01

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

  17. Strategic Planning for Chronic Disease Prevention in Rural America: Looking Through a PRISM Lens.

    PubMed

    Honeycutt, Amanda A; Wile, Kristina; Dove, Cassandra; Hawkins, Jackie; Orenstein, Diane

    2015-01-01

    Community-level strategic planning for chronic disease prevention. To share the outcomes of the strategic planning process used by Mississippi Delta stakeholders to prevent and reduce the negative impacts of chronic disease in their communities. A key component of strategic planning was participants' use of the Prevention Impacts Simulation Model (PRISM) to project the reduction, compared with the status quo, in deaths and costs from implementing interventions in Mississippi Delta communities. Participants in Mississippi Delta strategic planning meetings used PRISM, a user-friendly, evidence-based simulation tool that includes 22 categories of policy, systems, and environmental change interventions, to pose what-if questions that explore the likely short- and long-term effects of an intervention or any desired combination of the 22 categories of chronic disease intervention programs and policies captured in PRISM. These categories address smoking, air pollution, poor nutrition, and lack of physical activity. Strategic planning participants used PRISM outputs to inform their decisions and actions to implement interventions. Rural communities in the Mississippi Delta. A diverse group of 29 to 34 local chronic disease prevention stakeholders, known as the Mississippi Delta Strategic Alliance. Community plans and actions that were developed and implemented as a result of local strategic planning. Existing strategic planning efforts were complemented by the use of PRISM. The Mississippi Delta Strategic Alliance decided to implement new interventions to improve air quality and transportation and to expand existing interventions to reduce tobacco use and increase access to healthy foods. They also collaborated with the Department of Transportation to raise awareness and use of the current transportation network. The Mississippi Delta Strategic Alliance strategic planning process was complemented by the use of PRISM as a tool for strategic planning, which led to the

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

    NASA Astrophysics Data System (ADS)

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

    2012-04-01

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

  19. Strategic Implications of American Millennialism

    DTIC Science & Technology

    2008-05-22

    Strategic Implications of American Millennialism A Monograph by MAJOR Brian L. Stuckert U.S. Army School of Advanced Military Studies...Monograph 3. DATES COVERED (From - To) Sep 2007 – April 2008 4. TITLE AND SUBTITLE Strategic Implications of American Millennialism . 5a. CONTRACT NUMBER...Abstract 15. SUBJECT TERMS Strategic Implications, American Millennialism 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER

  20. Does strategic defense breed offense

    SciTech Connect

    York, H.

    1987-01-01

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

  1. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    Not Available

    1990-08-15

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

  2. Strategic Military Leaders - Leading Tomorrow

    DTIC Science & Technology

    2008-02-29

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

  3. Can Smog Raise Breast Cancer Risk?

    MedlinePlus

    ... gov/news/fullstory_164495.html Can Smog Raise Breast Cancer Risk? Exposure to fine-particle air pollution linked ... have dense breasts, a known risk factor for breast cancer, new research suggests. "It appears that women who ...

  4. 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)

  5. Could Parkinson's Disease Raise Stroke Risk?

    MedlinePlus

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

  6. Product Fund-Raising: A Practical Resource.

    ERIC Educational Resources Information Center

    Lemieux, Russell A.

    1997-01-01

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

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

    PubMed

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

    2017-02-07

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

  8. Reversed DNA strand cleavage specificity in initiation of Cre-LoxP recombination induced by the His289Ala active-site substitution.

    PubMed

    Gelato, Kathy A; Martin, Shelley S; Baldwin, Enoch P

    2005-11-25

    During the first steps of site-specific recombination, Cre protein cleaves and religates a specific homologous pair of LoxP strands to form a Holliday junction (HJ) intermediate. The HJ is resolved into recombination products through exchange of the second homologous strand pair. CreH289A, containing a His to Ala substitution in the conserved R-H-R catalytic motif, has a 150-fold reduced recombination rate and accumulates HJs. However, to produce these HJs, CreH289A exchanges the opposite set of strands compared to wild-type Cre (CreWT). To investigate how CreH289A and CreWT impose strand exchange order, we characterized their reactivities and strand cleavage preferences toward LoxP duplex and HJ substrates containing 8bp spacer substitutions. Remarkably, CreH289A had different and often opposite strand exchange preferences compared to CreWT with nearly all substrates. CreH289N was much less perturbed, implying that overall recombination rate and strand exchange depend more on His289 hydrogen bonding capability than on its acid/base properties. LoxP substitutions immediately 5' (S1 nucleotide) or 3' (S1' nucleotide) of the scissile phosphate had large effects on substrate utilization and strand exchange order. S1' substitutions, designed to alter base-unstacking events concomitant with Cre-induced LoxP bending, caused HJ accumulation and dramatically inverted the cleavage preferences. That pre-formed HJs were resolved via either strand in vitro suggests that inhibition of the "conformational switch" isomerization required to trigger the second strand exchange accounts for the observed HJ accumulation. Rather than reflecting CreWT behavior, CreH289A accumulates HJs of opposite polarity through a combination of its unique cleavage specificity and an HJ isomerization defect. The overall implication is that cleavage specificity is mediated by sequence-dependent DNA deformations that influence the scissile phosphate positioning and reactivity. A role of His289 may be to

  9. 75 FR 67695 - U.S. Strategic Command Strategic Advisory Group Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-03

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

  10. Enhancing the Strategic Capability of the Army: An Investigation of Strategic Thinking Tasks, Skills, and Development

    DTIC Science & Technology

    2016-02-01

    not fully supportive of strategic thinking development. Creating a climate in which important aspects of strategic thinking (e.g., reflection...practices could also be utilized to ensure selection boards value important strategic thinking developmental experiences. Further, strategic...candor) inhibit, rather than promote, strategic thinking development. Creating climates in which important aspects of strategic thinking (e.g

  11. 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.

  12. 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.

  13. Strategic Global Climate Command?

    NASA Astrophysics Data System (ADS)

    Long, J. C. S.

    2016-12-01

    Researchers have been exploring geoengineering because Anthropogenic GHG emissions could drive the globe towards unihabitability for people, wildlife and vegetation. Potential global deployment of these technologies is inherently strategic. For example, solar radiation management to reflect more sunlight might be strategically useful during a period of time where the population completes an effort to cease emissions and carbon removal technologies might then be strategically deployed to move the atmospheric concentrations back to a safer level. Consequently, deployment of these global technologies requires the ability to think and act strategically on the part of the planet's governments. Such capacity most definitely does not exist today but it behooves scientists and engineers to be involved in thinking through how global command might develop because the way they do the research could support the development of a capacity to deploy intervention rationally -- or irrationally. Internationalizing research would get countries used to working together. Organizing the research in a step-wise manner where at each step scientists become skilled at explaining what they have learned, the quality of the information they have, what they don't know and what more they can do to reduce or handle uncertainty, etc. Such a process can increase societal confidence in being able to make wise decisions about deployment. Global capacity will also be enhanced if the sceintific establishment reinvents misssion driven research so that the programs will identify the systemic issues invovled in any proposed technology and systematically address them with research while still encouraging individual creativity. Geoengineering will diverge from climate science in that geoengineering research needs to design interventions for some publically desirable goal and investigates whether a proposed intervention will acheive desired outcomes. The effort must be a systems-engineering design problem

  14. 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.

  15. Leading Strategic Leader Teams

    DTIC Science & Technology

    2008-03-25

    SUBTITLE Leading Strategic Leader Teams 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR( S ) Willard Burleson 5d...PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME( S ) AND ADDRESS(ES) U.S. Army War College ,122 Forbes Ave.,Carlisle...PA,17013-5220 8. PERFORMING ORGANIZATION REPORT NUMBER 9. SPONSORING/MONITORING AGENCY NAME( S ) AND ADDRESS(ES) 10. SPONSOR/MONITOR’S ACRONYM( S ) 11

  16. Strategic Posture Annex to the USAF Strategic Master Plan

    DTIC Science & Technology

    2015-05-01

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

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

    ERIC Educational Resources Information Center

    Leahey, Maureen; Wallace, Evie

    1988-01-01

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

  18. 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.

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

    PubMed

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

    2011-12-08

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

  20. A CRE1- regulated cluster is responsible for light dependent production of dihydrotrichotetronin in Trichoderma reesei.

    PubMed

    Monroy, Alberto Alonso; Stappler, Eva; Schuster, Andre; Sulyok, Michael; Schmoll, Monika

    2017-01-01

    Changing light conditions, caused by the rotation of earth resulting in day and night or growth on the surface or within a substrate, result in considerably altered physiological processes in fungi. For the biotechnological workhorse Trichoderma reesei, regulation of glycoside hydrolase gene expression, especially cellulase expression was shown to be a target of light dependent gene regulation. Analysis of regulatory targets of the carbon catabolite repressor CRE1 under cellulase inducing conditions revealed a secondary metabolite cluster to be differentially regulated in light and darkness and by photoreceptors. We found that this cluster is involved in production of trichodimerol and that the two polyketide synthases of the cluster are essential for biosynthesis of dihydrotrichotetronine (syn. bislongiquinolide or bisorbibutenolide). Additionally, an indirect influence on production of the peptaibol antibiotic paracelsin was observed. The two polyketide synthetase genes as well as the monooxygenase gene of the cluster were found to be connected at the level of transcription in a positive feedback cycle in darkness, but negative feedback in light, indicating a cellular sensing and response mechanism for the products of these enzymes. The transcription factor TR_102497/YPR2 residing within the cluster regulates the cluster genes in a light dependent manner. Additionally, an interrelationship of this cluster with regulation of cellulase gene expression was detected. Hence the regulatory connection between primary and secondary metabolism appears more widespread than previously assumed, indicating a sophisticated distribution of resources either to degradation of substrate (feed) or to antagonism of competitors (fight), which is influenced by light.

  1. 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

  2. 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

  3. Visualizing the lateral somitic frontier in the Prx1Cre transgenic mouse

    PubMed Central

    Durland, J Logan; Sferlazzo, Matteo; Logan, Malcolm; Burke, Ann Campbell

    2008-01-01

    Changes in the organization of the musculoskeletal system have accounted for many evolutionary adaptations in the vertebrate body plan. The musculoskeletal system develops from two mesodermal populations: somitic mesoderm gives rise to the axial skeleton and all of the skeletal muscle of the body, and lateral plate mesoderm gives rise to the appendicular skeleton. The recognition of embryonic domains resulting from the dynamics of morphogenesis has inspired new terminology based on developmental criteria. Two mesodermal domains are defined, primaxial and abaxial. The primaxial domain includes musculoskeletal structures comprising just somitic cells. The abaxial domain contains somitic myoblasts in connective tissue derived from lateral plate mesoderm, as well as lateral plate-derived skeletal structures. The boundary between these two domains is the lateral somitic frontier. Recent studies have described the developmental relationship between these two domains in the chick. In the present study, we describe the labelling pattern in the body of the Prx1/Cre/Z/AP compound transgenic mouse. The enhancer employed in this transgenic leads to reporter expression in the postcranial, somatic lateral plate mesoderm. The boundary between labelled and unlabelled cell populations is described at embryonic day (E)13.5 and E15.5. We argue that the distribution of labelled cells is consistent with the somatic lateral plate lineage, and therefore provides an estimate of the position of the lateral somitic frontier. The role of the frontier in both development and evolution is discussed. PMID:18430087

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

    PubMed

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

    2013-10-01

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

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

    PubMed Central

    Paredes, Silvana; Maggert, Keith A.

    2009-01-01

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

  6. Robust Cre-Mediated Recombination in Small Intestinal Stem Cells Utilizing the Olfm4 Locus

    PubMed Central

    Schuijers, Jurian; van der Flier, Laurens G.; van Es, Johan; Clevers, Hans

    2014-01-01

    Summary The epithelium of the small intestine is the most rapidly self-renewing tissue in mammals. We previously demonstrated the existence of a long-lived pool of cycling stem cells defined by Lgr5 expression at the bottom of intestinal crypts. An Lgr5-eGFP-IRES-CreERT2 knockin allele has been instrumental in characterizing and profiling these cells, yet its low level expression and its silencing in patches of adjacent crypts have not allowed quantitative gene deletion. Olfactomedin-4 (Olfm4) has emerged from a gene signature of Lgr5 stem cells as a robust marker for murine small intestinal stem cells. We observe that Olfm4null animals show no phenotype and report the generation of an Olfm4-IRES-eGFPCreERT2 knockin mouse model that allows visualization and genetic manipulation of Lgr5+ stem cells in the epithelium of the small intestine. The eGFPCreERT2 fusion protein faithfully marks all stem cells in the small intestine and induces the activation of a conditional LacZ reporter with robust efficiency. PMID:25254337

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

    SciTech Connect

    Canavan, G.H.

    1988-06-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  10. Strategic Decisions in Task-Oriented Reading.

    PubMed

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

    2015-12-28

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

  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. Healthcare Antibiotic Resistance Prevalence - DC (HARP-DC): A Regional Prevalence Assessment of Carbapenem-Resistant Enterobacteriaceae (CRE) in Healthcare Facilities in Washington, District of Columbia.

    PubMed

    Reuben, Jacqueline; Donegan, Nancy; Wortmann, Glenn; DeBiasi, Roberta; Song, Xiaoyan; Kumar, Princy; McFadden, Mary; Clagon, Sylvia; Mirdamadi, Janet; White, Diane; Harris, Jo Ellen; Browne, Angella; Hooker, Jane; Yochelson, Michael; Walker, Milena; Little, Gary; Jernigan, Gail; Hansen, Kathleen; Dockery, Brenda; Sinatro, Brendan; Blaylock, Morris; Harmon, Kimary; Iyengar, Preetha; Wagner, Trevor; Nelson, Jo Anne

    2017-08-01

    OBJECTIVE Carbapenem-resistant Enterobacteriaceae (CRE) are a significant clinical and public health concern. Understanding the distribution of CRE colonization and developing a coordinated approach are key components of control efforts. The prevalence of CRE in the District of Columbia is unknown. We sought to determine the CRE colonization prevalence within healthcare facilities (HCFs) in the District of Columbia using a collaborative, regional approach. DESIGN Point-prevalence study. SETTING This study included 16 HCFs in the District of Columbia: all 8 acute-care hospitals (ACHs), 5 of 19 skilled nursing facilities, 2 (both) long-term acute-care facilities, and 1 (the sole) inpatient rehabilitation facility. PATIENTS Inpatients on all units excluding psychiatry and obstetrics-gynecology. METHODS CRE identification was performed on perianal swab samples using real-time polymerase chain reaction, culture, and antimicrobial susceptibility testing (AST). Prevalence was calculated by facility and unit type as the number of patients with a positive result divided by the total number tested. Prevalence ratios were compared using the Poisson distribution. RESULTS Of 1,022 completed tests, 53 samples tested positive for CRE, yielding a prevalence of 5.2% (95% CI, 3.9%-6.8%). Of 726 tests from ACHs, 36 (5.0%; 95% CI, 3.5%-6.9%) were positive. Of 244 tests from long-term-care facilities, 17 (7.0%; 95% CI, 4.1%-11.2%) were positive. The relative prevalence ratios by facility type were 0.9 (95% CI, 0.5-1.5) and 1.5 (95% CI, 0.9-2.6), respectively. No CRE were identified from the inpatient rehabilitation facility. CONCLUSION A baseline CRE prevalence was established, revealing endemicity across healthcare settings in the District of Columbia. Our study establishes a framework for interfacility collaboration to reduce CRE transmission and infection. Infect Control Hosp Epidemiol 2017;38:921-929.

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

    PubMed

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

    2014-08-01

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

  14. Securitization of Russian Strategic Communication

    DTIC Science & Technology

    2010-12-02

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

  15. Strategic Management: A Comprehensive Bibliography.

    ERIC Educational Resources Information Center

    Chaffee, Ellen Earle; de Alba, Renee

    A bibliography on strategic management is presented to assist both practitioners and researchers. Criteria for inclusion were as follows: (1) general in scope, providing introductory information on a variety of subtopics within strategic management; (2) indications that the work is becoming a classic (i.e., frequent citations by other authors);…

  16. 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…

  17. 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…

  18. Strategic Planning for Higher Education.

    ERIC Educational Resources Information Center

    Kotler, Philip; Murphy, Patrick E.

    1981-01-01

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

  19. Strategic 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…

  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. The Science of Strategic Communication

    EPA Science Inventory

    The field of Strategic Communication involves a focused effort to identify, develop, and present multiple types of communication media on a given subject. A Strategic Communication program recognizes the limitations of the most common communication models (primarily “one s...

  2. Strategic Planning: A Participative Model.

    ERIC Educational Resources Information Center

    Weimer, Don; Jonas, Peter M.

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

  3. Army Environmental Cleanup Strategic Plan

    DTIC Science & Technology

    2009-05-01

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

  4. 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…

  5. Strategic Competence and Language Teaching.

    ERIC Educational Resources Information Center

    Rababah, Ghaleb Ahmed

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

  6. 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)

  7. 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…

  8. Strategic Planning and Information Systems.

    ERIC Educational Resources Information Center

    Shuman, Jack N.

    1982-01-01

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

  9. 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…

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

    PubMed Central

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

    2009-01-01

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

  11. 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.

  12. Successful strategic planning: creating clarity.

    PubMed

    Adams, Jim

    2005-01-01

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

  13. Tissue Specific Activation and Inactivation of the Neu Proto-Oncogene in Transgenic Mice Using Cre Recombinase

    DTIC Science & Technology

    2002-10-01

    now acquired the training and skills required to purify recombinant adenovirus and infect cells with an adenovirus containing Cre recombinase to... infection . Reportable Outcomes Papers Published Centrosome abnormalities, recurring deletions of chromosome 4, and genomic amplification of HER2/neu define...Differentiated maam. gl. marker 11.3 WAP 8.9 Glycaml Elevated in tumor model! 8.6 Glycoprotein 8.3 Connexin -30 Differentiation Marker 4.8 Connexin -26

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

    PubMed Central

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

    2015-01-01

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

  15. Use of Cre-adenovirus and CAR transgenic mice for efficient deletion of genes in post-thymic T cells

    PubMed Central

    Zha, Yuanyuan; Shah, Ramila; Locke, Frederick; Wong, Austin; Gajewski, Thomas F.

    2012-01-01

    Conditional gene deletion using lineage-specific transgenic expression of Cre has been useful for defining the role of specific gene products in mice in vivo. However, this technology has had limitations for studies of peripheral T cell biology, since the T-lineage promoters commonly used are active early in thymic development. As such, T cell development can be altered by the resulting genetic alterations, thus limiting the interpretation of the data in post-thymic T cell studies. Thus, new strategies are needed to delete targeted genes directly in peripheral T lymphocytes. The availability of transgenic mice expressing the CAR in the T cell compartment enabled testing of Cre-mediated recombination using an adenoviral vector in naïve peripheral T cells in vitro, even without cellular activation. Using Rosa26R reporter×CAR transgenic mice, we describe conditions by which Cre-mediated deletion of targeted genes can be achieved with primary T cells in vitro. These cells can also be adoptively transferred into defined recipient mice for study in vivo. We use conditional PTEN-deficient mice as proof of concept to confirm the value of this technique for deleting a negative regulator of T cell activation. This technology should be broadly applicable for studies of T cell specific gene deletion to gain understanding of function in the post-thymic T cell compartment. PMID:18177887

  16. Establishment of a Cre/loxP recombination system for N-terminal epitope tagging of genes in Tetrahymena

    PubMed Central

    2010-01-01

    Background Epitope tagging is a powerful strategy to study the function of proteins. Although tools for C-terminal protein tagging in the ciliated protozoan Tetrahymena thermophila have been developed, N-terminal protein tagging in this organism is still technically demanding. Results In this study, we have established a Cre/loxP recombination system in Tetrahymena and have applied this system for the N-terminal epitope tagging of Tetrahymena genes. Cre can be expressed in Tetrahymena and localizes to the macronucleus where it induces precise recombination at two loxP sequences in direct orientation in the Tetrahymena macronuclear chromosome. This Cre/loxP recombination can be used to remove a loxP-flanked drug-resistance marker from an N-terminal tagging construct after it is integrated into the macronucleus. Conclusions The system established in this study allows us to express an N-terminal epitope tagged gene from its own endogenous promoter in Tetrahymena. PMID:20626890

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

    PubMed

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

    2006-10-01

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

  18. A new mouse line for cell ablation by diphtheria toxin subunit A controlled by a Cre-dependent FLEx switch.

    PubMed

    Plummer, Nicholas W; Ungewitter, Erica K; Smith, Kathleen G; Yao, Humphrey H-C; Jensen, Patricia

    2017-09-05

    Recombinase responsive mouse lines expressing diphtheria toxin subunit A (DTA) are well established tools for targeted ablation of genetically defined cell populations. Here we describe a new knock-in allele at the Gt(Rosa)26Sor locus that retains the best features of previously described DTA alleles-including a CAG promoter, attenuated mutant DTA cDNA, and ubiquitous EGFP labeling-with the addition of a Cre-dependent FLEx switch for tight control of expression. The FLEx switch consists of two pairs of antiparallel lox sites requiring Cre-mediated recombination for inversion of the DTA to the proper orientation for transcription. We demonstrate its utility by Cre-dependent ablation of both a broad domain in the embryonic nervous system and a discrete population of cells in the fetal gonads. We conclude that this new DTA line is useful for targeted ablation of genetically-defined cell populations. This article is protected by copyright. All rights reserved. © 2017 Wiley Periodicals, Inc.

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

    PubMed Central

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

    2017-01-01

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

  20. 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.

  1. CRE: a cost effective and rapid approach for PCR-mediated concatenation of KRAS and EGFR exons

    PubMed Central

    Ramteke, Manoj P.; Patel, Kuldeep J; Godbole, Mukul; Vyas, Maulik; Karve, Kunal; Choughule, Anuradha; Prabhash, Kumar; Dutt, Amit

    2016-01-01

    Molecular diagnostics has changed the way lung cancer patients are treated worldwide. Of several different testing methods available, PCR followed by directed sequencing and amplification refractory mutation system (ARMS) are the two most commonly used diagnostic methods worldwide to detect mutations at  KRAS exon 2 and  EGFR kinase domain exons 18-21 in lung cancer. Compared to ARMS, the PCR followed by directed sequencing approach is relatively inexpensive but more cumbersome to perform. Moreover, with a limiting amount of genomic DNA from clinical formalin-fixed, paraffin-embedded (FFPE) specimens or fine biopsies of lung tumors, multiple rounds of PCR and sequencing reactions often get challenging. Here, we report a cost-effective single multiplex-PCR based method, CRE (for  Co-amplification of five  K RAS and  E GFR exons), followed by concatenation of the PCR product as a single linear fragment for direct sequencing. CRE is a robust protocol that can be adapted for routine use in clinical diagnostics with reduced variability, cost and turnaround time requiring a minimal amount of template DNA extracted from FFPE or fresh frozen tumor samples. As a proof of principle, CRE is able to detect the activating  EGFR L858R and T790M  EGFR mutations in lung cancer cell line and primary tumors. PMID:27127615

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

    PubMed

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

    2017-02-24

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

  3. Analysis of neural crest cell fate during cardiovascular development using Cre-activated lacZ/β-galactosidase staining.

    PubMed

    Zhang, Yanping; Ruest, L Bruno

    2012-01-01

    It is important to identify the mechanisms regulating cardiovascular development. However, complex genetic tools are often required, including transgenic animals that express the lacZ transgene encoding the β-galactosidase enzyme under the control of a specific promoter or following recombination with the Cre recombinase. The latter can be useful for identifying specific cell populations of the developing cardiovascular system, including neural crest cells. The tracking of these cells can help clarify their fate in mutant embryos and elucidate the etiology of some congenital cardiovascular birth defects. This chapter highlights the methods used to stain embryonic tissues in whole mount or sections to detect the expression of the lacZ transgene with a focus on tracking cardiac neural crest cells using the Wnt1-Cre and R26R mouse lines. We also provide a protocol using fluorescence-activated cell sorting for collecting neural crest cells for further analysis. These protocols can be used with any embryos expressing Cre and lacZ.

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

    PubMed

    Condie, Brian G; Urbanski, William M

    2014-01-01

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

  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. Strategic cycling: shaking complacency in healthcare strategic planning.

    PubMed

    Begun, J; Heatwole, K B

    1999-01-01

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

  7. A modified Cre-lox genetic switch to dynamically control metabolic flow in Saccharomyces cerevisiae.

    PubMed

    Yamanishi, Mamoru; Matsuyama, Takashi

    2012-05-18

    The control of metabolic flow is a prerequisite for efficient chemical production in transgenic microorganisms. Exogenous genes required for the biosynthesis of target chemicals are expressed under strong promoters, while the endogenous genes of the original metabolic pathway are repressed by disruption or mutation. These genetic manipulations occasionally cause harmful effects to the host. In the lactate-producing yeast Saccharomyces cerevisiae, where endogenous pyruvate decarboxylase (PDC) is disrupted and exogenous lactate dehydrogenase (LDH) is introduced, PDC deletion is extremely detrimental to cell growth but is required for efficient production of lactate. A suitable means to dynamically control the metabolic flow from ethanol fermentation during the growth phase to lactate fermentation during the production phase is needed. Here, we demonstrated that this flow can be controlled by the exclusive expression of PDC and LDH with a Cre-lox genetic switch. This switch was evaluated with a gene cassette that encoded two different fluorescence proteins and enabled changes in genotype and phenotype within 2 and 10 h, respectively. Transgenic yeast harboring this switch and the PDC-LDH cassette showed a specific growth rate (0.45 h (-1)) that was almost the same as that of wild-type (0.47 h (-1)). Upon induction of the genetic switch, the transgenic yeast produced lactate from up to 85.4% of the glucose substrate, while 91.7% of glucose went to ethanol before induction. We thus propose a "metabolic shift" concept that can serve as an alternative means to obtain gene products that are currently difficult to obtain by using conventional methodologies.

  8. Inactivation of Tgfbr2 in Osterix-Cre expressing Dental Mesenchyme Disrupts Molar Root Formation

    PubMed Central

    Coricor, George; MacDougall, Mary; Serra, Rosa

    2013-01-01

    It has been difficult to examine the role of TGF-ß in post-natal tooth development due to perinatal lethality in many of the signaling deficient mouse models. To address the role of Tgfbr2 in postnatal tooth development, we generated a mouse in which Tgfbr2 was deleted in odontoblast-and bone-producing mesenchyme. Osx-Cre;Tgfbr2fl/fl mice were generated (Tgfbr2cko) and postnatal tooth development was compared in Tgfbr2cko and control littermates. X-ray and μCT analysis showed that in Tgfbr2cko mice radicular dentin matrix density was reduced in the molars. Molar shape was abnormal and molar eruption was delayed in the mutant mice. Most significantly, defects in root formation, including failure of the root to elongate, were observed by postnatal day 10. Immunostaining for Keratin-14 (K14) was used to delineate Hertwig's epithelial root sheath (HERS). The results showed a delay in elongation and disorganization of the HERS in Tgfbr2cko mice. In addition, the HERS was maintained and the break up into epithelial rests was attenuated suggesting that Tgfbr2 acts on dental mesenchyme to indirectly regulate the formation and maintenance of the HERS. Altered odontoblast organization and reduced Dspp expression indicated that odontoblast differentiation was disrupted in the mutant mice likely contributing to the defect in root formation. Nevertheless, expression of Nfic, a key mesenchymal regulator of root development, was similar in Tgfbr2cko mice and controls. The number of osteoclasts in the bone surrounding the tooth was reduced and osteoblast differentiation was disrupted likely contributing to both root and eruption defects. We conclude that Tgfbr2 in dental mesenchyme and bone is required for tooth development particularly root formation. PMID:23933490

  9. Inactivation of Tgfbr2 in Osterix-Cre expressing dental mesenchyme disrupts molar root formation.

    PubMed

    Wang, Ying; Cox, Megan K; Coricor, George; MacDougall, Mary; Serra, Rosa

    2013-10-01

    It has been difficult to examine the role of TGF-ß in post-natal tooth development due to perinatal lethality in many of the signaling deficient mouse models. To address the role of Tgfbr2 in postnatal tooth development, we generated a mouse in which Tgfbr2 was deleted in odontoblast- and bone-producing mesenchyme. Osx-Cre;Tgfbr2(fl/fl) mice were generated (Tgfbr2(cko)) and post-natal tooth development was compared in Tgfbr2(cko) and control littermates. X-ray and μCT analysis showed that in Tgfbr2(cko) mice radicular dentin matrix density was reduced in the molars. Molar shape was abnormal and molar eruption was delayed in the mutant mice. Most significantly, defects in root formation, including failure of the root to elongate, were observed by postnatal day 10. Immunostaining for Keratin-14 (K14) was used to delineate Hertwig's epithelial root sheath (HERS). The results showed a delay in elongation and disorganization of the HERS in Tgfbr2(cko) mice. In addition, the HERS was maintained and the break up into epithelial rests was attenuated suggesting that Tgfbr2 acts on dental mesenchyme to indirectly regulate the formation and maintenance of the HERS. Altered odontoblast organization and reduced Dspp expression indicated that odontoblast differentiation was disrupted in the mutant mice likely contributing to the defect in root formation. Nevertheless, expression of Nfic, a key mesenchymal regulator of root development, was similar in Tgfbr2(cko) mice and controls. The number of osteoclasts in the bone surrounding the tooth was reduced and osteoblast differentiation was disrupted likely contributing to both root and eruption defects. We conclude that Tgfbr2 in dental mesenchyme and bone is required for tooth development particularly root formation. © 2013 Elsevier Inc. All rights reserved.

  10. Tamoxifen dosing for Cre-mediated recombination in experimental bronchopulmonary dysplasia.

    PubMed

    Ruiz-Camp, Jordi; Rodríguez-Castillo, José Alberto; Herold, Susanne; Mayer, Konstantin; Vadász, István; Tallquist, Michelle D; Seeger, Werner; Ahlbrecht, Katrin; Morty, Rory E

    2017-02-01

    Bronchopulmonary dysplasia (BPD) is the most common complication of preterm birth characterized by blunted post-natal lung development. BPD can be modelled in mice by exposure of newborn mouse pups to elevated oxygen levels. Little is known about the mechanisms of perturbed lung development associated with BPD. The advent of transgenic mice, where genetic rearrangements can be induced in particular cell-types at particular time-points during organogenesis, have great potential to explore the pathogenic mechanisms at play during arrested lung development. Many inducible, conditional transgenic technologies available rely on the application of the estrogen-receptor modulator, tamoxifen. While tamoxifen is well-tolerated and has been widely employed in adult mice, or in healthy developing mice; tamoxifen is not well-tolerated in combination with hyperoxia, in the most widely-used mouse model of BPD. To address this, we set out to establish a safe and effective tamoxifen dosing regimen that can be used in newborn mouse pups subjected to injurious stimuli, such as exposure to elevated levels of environmental oxygen. Our data reveal that a single intraperitoneal dose of tamoxifen of 0.2 mg applied to newborn mouse pups in 10 μl Miglyol vehicle was adequate to successfully drive Cre recombinase-mediated genome rearrangements by the fifth day of life, in a murine model of BPD. The number of recombined cells was comparable to that observed in regular tamoxifen administration protocols. These findings will be useful to investigators where tamoxifen dosing is problematic in the background of injurious stimuli and mouse models of human and veterinary disease.

  11. Conventional Strategic Deterrence

    SciTech Connect

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

    1992-08-01

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

  12. 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.

  13. 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…

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

    PubMed

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

    2017-03-10

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

  15. 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…

  16. 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…

  17. 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…

  18. 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…

  19. People on the Farm: Raising Beef Cattle.

    ERIC Educational Resources Information Center

    Crain, Robert L.

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

  20. Raising Environmental Awareness among Miners in Iran

    ERIC Educational Resources Information Center

    Mozaffari, Ezatollah

    2013-01-01

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

  1. 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. YOU CAN RAISE YOUR HANDICAPPED CHILD.

    ERIC Educational Resources Information Center

    AYRAULT, EVELYN WEST

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

  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…

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

    ERIC Educational Resources Information Center

    Ransome, Whitney, Ed.; And Others

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

  5. National Organizations Raising Public Consciousness. Part II.

    ERIC Educational Resources Information Center

    Exceptional Parent, 1980

    1980-01-01

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

  6. 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. Pragmatics, Awareness Raising, and the Cooperative Principle

    ERIC Educational Resources Information Center

    Murray, Neil

    2010-01-01

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

  8. Solo Parenting: Raising Strong & Happy Families.

    ERIC Educational Resources Information Center

    Chambers, Diane

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

  9. YOU CAN RAISE YOUR HANDICAPPED CHILD.

    ERIC Educational Resources Information Center

    AYRAULT, EVELYN WEST

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

  10. Pragmatics, Awareness Raising, and the Cooperative Principle

    ERIC Educational Resources Information Center

    Murray, Neil

    2010-01-01

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

  11. 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…

  12. Some Blunt Talk about Fund Raising.

    ERIC Educational Resources Information Center

    Buchanan, Peter McE.

    1991-01-01

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

  13. 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…

  14. 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…

  15. Raising the Career Aspirations of Gifted Girls.

    ERIC Educational Resources Information Center

    Kerr, Barbara A.

    1983-01-01

    Tested the effectiveness of a one-day career guidance laboratory on the career aspirations of 48 gifted male and female high school students. Results showed the program was effective for the girls, who raised their career aspirations significantly while the boys, who began with higher aspirations, did not change. (JAC)

  16. The Power of the Raised Eyebrow

    ERIC Educational Resources Information Center

    Burton, Grace M.

    1977-01-01

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

  17. Raised Water Tables Affect Southern Hardwood Growth

    Treesearch

    W. M. Broadfoot

    1973-01-01

    In natural stands near Demopolis Lock and Dam Reservoir in Alabama, the average growth in tree radius increased about 50 percent in the 5 years after the water table was raised from an indefinite depth to within reach of the tree roots. In natural stands near the Jim Woodruff Reservoir in Florida, radial growth of trees also increased markedly after the water table was...

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

    ERIC Educational Resources Information Center

    Ransome, Whitney, Ed.; And Others

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

  19. 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…

  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. Raising Children Bilingually Is Hard: Why Bother?

    ERIC Educational Resources Information Center

    Sims, Margaret; Ellis, Elizabeth M.

    2014-01-01

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

  2. Raising Children Bilingually Is Hard: Why Bother?

    ERIC Educational Resources Information Center

    Sims, Margaret; Ellis, Elizabeth M.

    2014-01-01

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

  3. Solo Parenting: Raising Strong & Happy Families.

    ERIC Educational Resources Information Center

    Chambers, Diane

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

  4. 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…

  5. 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…

  6. 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…

  7. Issues Raised by the Follow Through Evaluation.

    ERIC Educational Resources Information Center

    House, Ernest R.; Hutchins, Elizabeth J.

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

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

    PubMed Central

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

    2016-01-01

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

  9. 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

  10. 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

  11. Strategic planning in diagnostic imaging: meeting the challenge.

    PubMed

    Lexa, Frank J

    2008-02-01

    The 21st century has raised new and significant challenges to the practice of diagnostic imaging. Radiologists will have to navigate a difficult path as they face threats from disruptive technologies, adverse demographic changes, pressures to limit reimbursement both from the public and from the private sectors, increased domestic and crossborder competition, and many others. Success in these trying times will require greater attention to strategic planning if we are to thrive and survive in radiology. Strategic planning and tactical implementation methodologies were reviewed, from Sun Tzu to the present day, for applicability to the needs of modern radiology groups. A framework for developing and implementing strategic plans was constructed to assist radiology leaders and groups in considering the spectrum of tasks, from gathering intelligence, to developing scenarios, to implementing and evaluating tactical plans. Strategy and tactics are too important to be ignored or left to others. They need to be core activities for all radiology leaders. Frameworks can be used to help in providing structure and rigour to strategic planning efforts at the department and group level.

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

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

  13. The Value of Strategic Partnerships

    ScienceCinema

    Gould, Josh; Narayan, Amit; McNutt, Ty

    2016-07-12

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

  14. 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.

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

    PubMed

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

    2015-12-01

    The dihydrofolate reductase gene amplification system is widely used in Chinese hamster ovary (CHO) cells for the industrial production of therapeutic proteins. To enhance the efficiency of conventional gene amplification systems, we previously presented a novel method using cell-cycle checkpoint engineering. Here, we constructed high-producing and stable cells by the conditional expression of mutant cell division cycle 25 homolog B (CDC25B) using the Cre-loxP system. A bispecific antibody-producing CHO DG44-derived cell line was transfected with floxed mutant CDC25B. After inducing gene amplification in the presence of 250 nM methotrexate, mutant CDC25B sequence was removed by Cre recombinase protein expression. Overexpression of the floxed mutant CDC25B significantly enhanced the efficiency of transgene amplification and productivity. Moreover, the specific production rate of the isolated clone CHO Cre-1 and Cre-2 were approximately 11-fold and 15-fold higher than that of mock-transfected clone CHO Mock-S. Chromosomal aneuploidy was increased by mutant CDC25B overexpression, but Cre-1 and Cre-2 did not show any changes in chromosome number during long-term cultivation, as is the case with CHO Mock-S. Our results suggest that high-producing and stable cells can be constructed by conditionally controlling a cell-cycle checkpoint integrated in conventional gene amplification systems. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  16. Elevated cJUN expression and an ATF/CRE site within the ATF3 promoter contribute to activation of ATF3 transcription by the amino acid response.

    PubMed

    Fu, Lingchen; Kilberg, Michael S

    2013-02-15

    Mammalian cells respond to amino acid deprivation through multiple signaling pathways referred to as the amino acid response (AAR). Transcription factors mediate the AAR after their activation by several mechanisms; examples include translational control (activating transcription factor 4, ATF4), phosphorylation (p-cJUN), and transcriptional control (ATF3). ATF4 induces ATF3 transcription through a promoter-localized C/EBP-ATF response element (CARE). The present report characterizes an ATF/CRE site upstream of the CARE that also contributes to AAR-induced ATF3 transcription. ATF4 binds to the ATF/CRE and CARE sequences and both are required for a maximal response to ATF4 induction. ATF3, which antagonizes ATF4 and represses its own gene, also exhibited binding activity to the ATF/CRE and CARE sequences. The AAR resulted in elevated total cJUN and p-cJUN protein levels and both forms exhibited binding activity to the ATF/CRE and CARE ATF3 sequences. Knockdown of AAR-enhanced cJUN expression blocked induction of the ATF3 gene and mutation of either the ATF/CRE or the CARE site prevented the cJUN-dependent increase in ATF3-driven luciferase activity. The results indicate that both increased cJUN and the cis-acting ATF/CRE sequence within the ATF3 promoter contribute to the transcriptional activation of the gene during the AAR.

  17. Elevated cJUN expression and an ATF/CRE site within the ATF3 promoter contribute to activation of ATF3 transcription by the amino acid response

    PubMed Central

    Fu, Lingchen

    2013-01-01

    Mammalian cells respond to amino acid deprivation through multiple signaling pathways referred to as the amino acid response (AAR). Transcription factors mediate the AAR after their activation by several mechanisms; examples include translational control (activating transcription factor 4, ATF4), phosphorylation (p-cJUN), and transcriptional control (ATF3). ATF4 induces ATF3 transcription through a promoter-localized C/EBP-ATF response element (CARE). The present report characterizes an ATF/CRE site upstream of the CARE that also contributes to AAR-induced ATF3 transcription. ATF4 binds to the ATF/CRE and CARE sequences and both are required for a maximal response to ATF4 induction. ATF3, which antagonizes ATF4 and represses its own gene, also exhibited binding activity to the ATF/CRE and CARE sequences. The AAR resulted in elevated total cJUN and p-cJUN protein levels and both forms exhibited binding activity to the ATF/CRE and CARE ATF3 sequences. Knockdown of AAR-enhanced cJUN expression blocked induction of the ATF3 gene and mutation of either the ATF/CRE or the CARE site prevented the cJUN-dependent increase in ATF3-driven luciferase activity. The results indicate that both increased cJUN and the cis-acting ATF/CRE sequence within the ATF3 promoter contribute to the transcriptional activation of the gene during the AAR. PMID:23269699

  18. Nestin-Cre mice are affected by hypopituitarism, which is not due to significant activity of the transgene in the pituitary gland.

    PubMed

    Galichet, Christophe; Lovell-Badge, Robin; Rizzoti, Karine

    2010-07-06

    Nestin-Cre mice express Cre recombinase under control of the rat nestin promoter and central nervous system (CNS) enhancer. While endogenous Nestin is expressed in some other tissues including the pituitary gland, Nestin-Cre mice induce recombination predominantly in the CNS. For this reason, they have been widely used to explore gene function or cell fate in the latter. Pituitary hormonal deficiencies, or hypopituitarism, are associated with a wide range of symptoms and with a significant morbidity. These can have a neural and/or a pituitary origin as the gland's secretions are controlled by the hypothalamus. We report here that Nestin-Cre mice themselves are affected by mild hypopituitarism. Hence, physiological consequences are expected, especially in combination with defects resulting from Cre mediated deletion of any gene under investigation. To further investigate the origin of this phenotype, we re-examined the activity of the transgene. We compared it with expression of Nestin itself in the context of the hypothalamo-pituitary axis, especially in the light of a recent report showing pituitary Nestin-Cre activity, which contrasts with previous data. Our results disagree with those of this recent study and do not support the claim that Nestin positive cells are present in the pituitary anlagen, the Rathke's pouch (RP). Moreover we did not observe any significant activity in the post-natal pituitary, in agreement with the initial report.

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

    SciTech Connect

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

    1994-10-01

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

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

    PubMed

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

    2014-05-02

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