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Sample records for ability pathways rap

  1. A Preliminary Outcome Study of Response Ability Pathways Training

    ERIC Educational Resources Information Center

    Forthun, Larry F.; McCombie, Jeff W.

    2007-01-01

    Approximately 68 classroom teachers participated in a preliminary evaluation of Response Ability Pathways (RAP), a reclaiming training course for adults who work with children and youth. RAP offers basic training in the Circle of Courage Model and provides participants with general strategies for assisting youth who are experiencing challenges.…

  2. Response Ability Pathways: A Curriculum for Connecting

    ERIC Educational Resources Information Center

    Koehler, Nancy; Seger, Vikki

    2005-01-01

    This article describes a new training curriculum for educators, youth workers, and mentors which draws from research and best practices in positive youth development and positive behavior support. Response Ability Pathways or RAP focuses on three practical interventions: connect to others for support, clarify challenging problems, and restore…

  3. Increased sugar uptake promotes oncogenesis via EPAC/RAP1 and O-GlcNAc pathways

    PubMed Central

    Onodera, Yasuhito; Nam, Jin-Min; Bissell, Mina J.

    2013-01-01

    There is a considerable resurgence of interest in the role of aerobic glycolysis in cancer; however, increased glycolysis is frequently viewed as a consequence of oncogenic events that drive malignant cell growth and survival. Here we provide evidence that increased glycolytic activation itself can be an oncogenic event in a physiologically relevant 3D culture model. Overexpression of glucose transporter type 3 (GLUT3) in nonmalignant human breast cells activated known oncogenic signaling pathways, including EGFR, β1 integrin, MEK, and AKT, leading to loss of tissue polarity and increased growth. Conversely, reduction of glucose uptake in malignant cells promoted the formation of organized and growth-arrested structures with basal polarity, and suppressed oncogenic pathways. Unexpectedly and importantly, we found that unlike reported literature, in 3D the differences between “normal” and malignant phenotypes could not be explained by HIF-1α/2α, AMPK, or mTOR pathways. Loss of epithelial integrity involved activation of RAP1 via exchange protein directly activated by cAMP (EPAC), involving also O-linked N-acetylglucosamine modification downstream of the hexosamine biosynthetic pathway. The former, in turn, was mediated by pyruvate kinase M2 (PKM2) interaction with soluble adenylyl cyclase. Our findings show that increased glucose uptake activates known oncogenic pathways to induce malignant phenotype, and provide possible targets for diagnosis and therapeutics. PMID:24316969

  4. Prostaglandin E2 regulates renal cell carcinoma invasion through the EP4 receptor-Rap GTPase signal transduction pathway.

    PubMed

    Wu, Juanjuan; Zhang, Yushan; Frilot, Nicole; Kim, Jae I; Kim, Wan-Ju; Daaka, Yehia

    2011-09-30

    Prognosis for patients with early stage kidney cancer has improved, but the treatment options for patients with locally advanced disease and metastasis remain few. Understanding the molecular mechanisms that regulate invasion and metastasis is critical for developing successful therapies to treat these patients. Proinflammatory prostaglandin E(2) plays an important role in cancer initiation and progression via activation of cognate EP receptors that belong to the superfamily of G protein-coupled receptors. Here we report that prostaglandin E(2) promotes renal cancer cell invasion through a signal transduction pathway that encompasses EP4 and small GTPase Rap. Inactivation of Rap signaling with Rap1GAP, like inhibition of EP4 signaling with ligand antagonist or knockdown with shRNA, reduces the kidney cancer cell invasion. Human kidney cells evidence increased EP4 and decreased Rap1GAP expression levels in the malignant compared with benign samples. These results support the idea that targeted inhibition of EP4 signaling and restoration of Rap1GAP expression constitute a new strategy to control kidney cancer progression.

  5. Prostacyclin regulates bone growth via the Epac/Rap1 pathway.

    PubMed

    Hutchison, Michele R; White, Perrin C

    2015-02-01

    Prostaglandins, particularly PGE2, are important to adult bone and joint health, but how prostaglandins act on growth plate cartilage to affect bone growth is unclear. We show that growth plate cartilage is distinct from articular cartilage with respect to cyclooxygenase (COX)-2 mRNA expression; although articular chondrocytes express very little COX-2, COX-2 expression is high in growth plate chondrocytes and is increased by IGF-I. In bovine primary growth plate chondrocytes, ATDC5 cells, and human metatarsal explants, inhibition of COX activity with nonsteroidal antiinflammatory drugs (NSAIDs) inhibits chondrocyte proliferation and ERK activation by IGF-I. This inhibition is reversed by prostaglandin E2 and prostacyclin (PGI2) but not by prostaglandin D2 or thromboxane B2. Inhibition of COX activity in young mice by ip injections of NSAIDs causes dwarfism. In growth plate chondrocytes, inhibition of proliferation and ERK activation by NSAIDs is reversed by forskolin, 8-bromoadenosine, 3',5'-cAMP and a prostacyclin analog, iloprost. The inhibition of proliferation and ERK activation by celecoxib is also reversed by 8CPT-2Me-cAMP, an activator of Epac, implicating the small G protein Rap1 in the pathway activated by iloprost. These results imply that prostacyclin is required for proper growth plate development and bone growth. PMID:25406016

  6. Rap-Interacting Proteins are Key Players in the Rap Symphony Orchestra.

    PubMed

    Guo, Xiao-Xi; An, Su; Yang, Yang; Liu, Ying; Hao, Qian; Xu, Tian-Rui

    2016-01-01

    Rap, a member of the Ras-like small G-protein family, is a key node among G-protein coupled receptors (GPCR), receptor tyrosine kinases (RTKs), ion channels and many other downstream pathways. Rap plays a unique role in cell morphogenesis, adhesion, migration, exocytosis, proliferation, apoptosis and carcinogenesis. The complexity and diversity of Rap functions are tightly regulated by Rap-interacting proteins such as GEFs, GAPs, Rap effectors and scaffold proteins. These interacting proteins decide the subcellular localization of Rap, the interaction modes with downstream Rap effectors and tune Rap as an atypical molecular conductor, coupling extra- and intracellular signals to various pathways. In this review, we summarize four groups of Rap-interacting proteins, highlight their distinctions in Rap-binding properties and interactive modes and discuss their contribution to the spatiotemporal regulation of Rap as well as the implications of targeting Rap-interacting proteins in human cancer therapy.

  7. Rap-Interacting Proteins are Key Players in the Rap Symphony Orchestra.

    PubMed

    Guo, Xiao-Xi; An, Su; Yang, Yang; Liu, Ying; Hao, Qian; Xu, Tian-Rui

    2016-01-01

    Rap, a member of the Ras-like small G-protein family, is a key node among G-protein coupled receptors (GPCR), receptor tyrosine kinases (RTKs), ion channels and many other downstream pathways. Rap plays a unique role in cell morphogenesis, adhesion, migration, exocytosis, proliferation, apoptosis and carcinogenesis. The complexity and diversity of Rap functions are tightly regulated by Rap-interacting proteins such as GEFs, GAPs, Rap effectors and scaffold proteins. These interacting proteins decide the subcellular localization of Rap, the interaction modes with downstream Rap effectors and tune Rap as an atypical molecular conductor, coupling extra- and intracellular signals to various pathways. In this review, we summarize four groups of Rap-interacting proteins, highlight their distinctions in Rap-binding properties and interactive modes and discuss their contribution to the spatiotemporal regulation of Rap as well as the implications of targeting Rap-interacting proteins in human cancer therapy. PMID:27322838

  8. RAP1-mediated MEK/ERK pathway defects in Kabuki syndrome.

    PubMed

    Bögershausen, Nina; Tsai, I-Chun; Pohl, Esther; Kiper, Pelin Özlem Simsek; Beleggia, Filippo; Percin, E Ferda; Keupp, Katharina; Matchan, Angela; Milz, Esther; Alanay, Yasemin; Kayserili, Hülya; Liu, Yicheng; Banka, Siddharth; Kranz, Andrea; Zenker, Martin; Wieczorek, Dagmar; Elcioglu, Nursel; Prontera, Paolo; Lyonnet, Stanislas; Meitinger, Thomas; Stewart, A Francis; Donnai, Dian; Strom, Tim M; Boduroglu, Koray; Yigit, Gökhan; Li, Yun; Katsanis, Nicholas; Wollnik, Bernd

    2015-09-01

    The genetic disorder Kabuki syndrome (KS) is characterized by developmental delay and congenital anomalies. Dominant mutations in the chromatin regulators lysine (K)-specific methyltransferase 2D (KMT2D) (also known as MLL2) and lysine (K)-specific demethylase 6A (KDM6A) underlie the majority of cases. Although the functions of these chromatin-modifying proteins have been studied extensively, the physiological systems regulated by them are largely unknown. Using whole-exome sequencing, we identified a mutation in RAP1A that was converted to homozygosity as the result of uniparental isodisomy (UPD) in a patient with KS and a de novo, dominant mutation in RAP1B in a second individual with a KS-like phenotype. We elucidated a genetic and functional interaction between the respective KS-associated genes and their products in zebrafish models and patient cell lines. Specifically, we determined that dysfunction of known KS genes and the genes identified in this study results in aberrant MEK/ERK signaling as well as disruption of F-actin polymerization and cell intercalation. Moreover, these phenotypes could be rescued in zebrafish models by rebalancing MEK/ERK signaling via administration of small molecule inhibitors of MEK. Taken together, our studies suggest that the KS pathophysiology overlaps with the RASopathies and provide a potential direction for treatment design. PMID:26280580

  9. RAP1-mediated MEK/ERK pathway defects in Kabuki syndrome

    PubMed Central

    Bögershausen, Nina; Tsai, I-Chun; Pohl, Esther; Kiper, Pelin Özlem Simsek; Beleggia, Filippo; Percin, E. Ferda; Keupp, Katharina; Matchan, Angela; Milz, Esther; Alanay, Yasemin; Kayserili, Hülya; Liu, Yicheng; Banka, Siddharth; Kranz, Andrea; Zenker, Martin; Wieczorek, Dagmar; Elcioglu, Nursel; Prontera, Paolo; Lyonnet, Stanislas; Meitinger, Thomas; Stewart, A. Francis; Donnai, Dian; Strom, Tim M.; Boduroglu, Koray; Yigit, Gökhan; Li, Yun; Katsanis, Nicholas; Wollnik, Bernd

    2015-01-01

    The genetic disorder Kabuki syndrome (KS) is characterized by developmental delay and congenital anomalies. Dominant mutations in the chromatin regulators lysine (K)–specific methyltransferase 2D (KMT2D) (also known as MLL2) and lysine (K)–specific demethylase 6A (KDM6A) underlie the majority of cases. Although the functions of these chromatin-modifying proteins have been studied extensively, the physiological systems regulated by them are largely unknown. Using whole-exome sequencing, we identified a mutation in RAP1A that was converted to homozygosity as the result of uniparental isodisomy (UPD) in a patient with KS and a de novo, dominant mutation in RAP1B in a second individual with a KS-like phenotype. We elucidated a genetic and functional interaction between the respective KS-associated genes and their products in zebrafish models and patient cell lines. Specifically, we determined that dysfunction of known KS genes and the genes identified in this study results in aberrant MEK/ERK signaling as well as disruption of F-actin polymerization and cell intercalation. Moreover, these phenotypes could be rescued in zebrafish models by rebalancing MEK/ERK signaling via administration of small molecule inhibitors of MEK. Taken together, our studies suggest that the KS pathophysiology overlaps with the RASopathies and provide a potential direction for treatment design. PMID:26280580

  10. Integrating RAP in Public Schools: Successes and Challenges

    ERIC Educational Resources Information Center

    Shields, Julie; Milstein, Mindy; Posner, Sandi Ives

    2010-01-01

    For students with emotional disability (ED), school can be a stressful experience marked by disapproval, rejection, isolation, and shame. Staff serving these students in a large school district received Response Ability Pathways (RAP) training. This article summarizes positive impacts on behavior, as well as ongoing challenges in changing school…

  11. Creating the Infrastructure for Organizational Change with RAP

    ERIC Educational Resources Information Center

    Shields, Julie; Milstein, Mindy; Robinson, Consuela

    2012-01-01

    In order to thrive, organizations must undergo significant change at various points in their development. Such is the case with Montgomery County Public Schools (MCPS) Emotional Disability Services in the beginning process of implementing Response Ability Pathways (RAP) with staff and students. The impetus for change originated from an…

  12. miR-9 and miR-124 synergistically affect regulation of dendritic branching via the AKT/GSK3β pathway by targeting Rap2a

    PubMed Central

    Xue, Qian; Yu, Caiyong; Wang, Yan; Liu, Ling; Zhang, Kun; Fang, Chao; Liu, Fangfang; Bian, Ganlan; Song, Bing; Yang, Angang; Ju, Gong; Wang, Jian

    2016-01-01

    A single microRNA (miRNA) can regulate expression of multiple proteins, and expression of an individual protein may be controlled by numerous miRNAs. This regulatory pattern strongly suggests that synergistic effects of miRNAs play critical roles in regulating biological processes. miR-9 and miR-124, two of the most abundant miRNAs in the mammalian nervous system, have important functions in neuronal development. In this study, we identified the small GTP-binding protein Rap2a as a common target of both miR-9 and miR-124. miR-9 and miR-124 together, but neither miRNA alone, strongly suppressed Rap2a, thereby promoting neuronal differentiation of neural stem cells (NSCs) and dendritic branching of differentiated neurons. Rap2a also diminished the dendritic complexity of mature neurons by decreasing the levels of pAKT and pGSK3β. Our results reveal a novel pathway in which miR-9 and miR-124 synergistically repress expression of Rap2a to sustain homeostatic dendritic complexity during neuronal development and maturation. PMID:27221778

  13. Plk2 Raps up Ras to subdue synapses

    PubMed Central

    Lee, Kea Joo; Hoe, Hyang-Sook

    2011-01-01

    We recently identified the activity-inducible protein kinase Plk2 as a novel overseer of the balance between Ras and Rap small GTPases. Plk2 achieves a profound level of regulatory control by interacting with and phosphorylating at least four Ras and Rap guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs). Combined, these actions result in synergistic suppression of Ras and hyperstimulation of Rap signaling. Perturbation of Plk2 function abolished homeostatic adaptation of synapses to enhanced activity and impaired behavioral adaptation in various learning tasks, indicating that this regulation was critical for maintaining appropriate Ras/Rap levels. These studies provide insights into the highly cooperative nature of Ras and Rap regulation in neurons. However, different GEF and GAP substrates of Plk2 also controlled specific aspects of dendritic spine morphology, illustrating the ability of individual GAPs/GEFs to assemble microdomains of Ras and Rap signaling that respond to different stimuli and couple to distinct output pathways. PMID:21776418

  14. Dysfunction of the PI3 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia.

    PubMed

    Moore, Samantha F; Hunter, Roger W; Harper, Matthew T; Savage, Joshua S; Siddiq, Samreen; Westbury, Sarah K; Poole, Alastair W; Mumford, Andrew D; Hers, Ingeborg

    2013-02-14

    Patients with myeloproliferative disorders (MPDs), such as essential thrombocythemia (ET) have increased risk of thrombosis and bleeding, which are major sources of morbidity and mortality. Most MPD patients have a gain of function mutation in Janus kinase 2 (JAK2V617F), but little is known how JAK2V617F affects platelet function. Here, we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin (which couples to JAK2) on this pathway. In contrast, SFLLRN-mediated P-selectin expression, ATP secretion, phosphorylation of the PKC substrate pleckstrin, and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets. In addition, thrombopoietin-mediated JAK2 phosphorylation was unchanged, suggesting that signaling pathways activated downstream of JAK2 are impaired. Indeed, we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the PI3 kinase substrate Akt, and have reduced activation of Rap1 in response to thrombopoietin, IGF-1,ADP, SFLLRN, and thrombin. This effect was independent of Giα P2Y12 purinergic receptor function as ADP-mediated inhibition of VASP phosphorylation was unchanged. These results demonstrate that the PI3 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients, resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation. PMID:23243278

  15. Early activation of the Kaposi's sarcoma-associated herpesvirus RTA, RAP, and MTA promoters by the tetradecanoyl phorbol acetate-induced AP1 pathway.

    PubMed

    Wang, Shizhen Emily; Wu, Frederick Y; Chen, Honglin; Shamay, Meir; Zheng, Qizhi; Hayward, Gary S

    2004-04-01

    Kaposi's sarcoma-associated herpesvirus (KSHV) maintains a latent infection in primary effusion lymphoma (PEL) cells, but treatment with tetradecanoyl phorbol acetate (TPA) can trigger the full lytic-cycle replication in some of these cells. During lytic-cycle replication, the KSHV-encoded replication and transcription activator (RTA or ORF50), the mRNA transport and accumulation protein (MTA), and the replication-associated protein (RAP) all play crucial roles in expression of downstream viral genes as well as in mediation of viral DNA replication. The cellular CCAAT/enhancer-binding protein alpha (C/EBP alpha) is induced in TPA-treated PEL cells and contributes to transactivation of the promoters for all of these genes through both direct binding and cooperative interactions with RTA and RAP targeted to upstream C/EBP sites. However, little is known about how RTA expression is triggered initially at the earliest stages after TPA induction when the C/EBP alpha levels are still limited. Treatment with TPA proved to significantly induce both AP1 DNA-binding activity and levels of activated phosphorylated cJUN in PEL cells and ectopic expression of cJUN-plus-cFOS-induced RTA protein expression in PEL cells. Cotransfected cJUN plus cFOS or TPA treatment transactivated the KSHV RTA, RAP, and MTA promoters in an AP1-binding site-dependent manner in all three promoters. Chromatin immunoprecipitation assays confirmed that cJUN associates with these KSHV target promoters in PEL cells as early as 4 h after TPA treatment. Furthermore, the KSHV RTA and RAP proteins both interact with cJUN or both cJUN and cFOS in vitro or by coimmunoprecipitation from induced PEL cells and enhance cJUN-plus-cFOS-mediated transactivation of these viral promoters. Both increased phosphorylated cJUN and AP1 DNA-binding activity was detected as early as 1 h after TPA treatment in PEL cells, suggesting that AP1 activity may be crucial for very early activation of the RAP, MTA, and RTA promoters

  16. Regulating Rap small G-proteins in time and space.

    PubMed

    Gloerich, Martijn; Bos, Johannes L

    2011-10-01

    Signaling by the small G-protein Rap is under tight regulation by its GEFs and GAPs. These are multi-domain proteins that are themselves controlled by distinct upstream pathways, and thus couple different extra- and intracellular cues to Rap. The individual RapGEFs and RapGAPs are, in addition, targeted to specific cellular locations by numerous anchoring mechanisms and, consequently, may control different pools of Rap. Here, we review the various activating signals and targeting mechanisms of these proteins and discuss their contribution to the spatiotemporal regulation and biological functions of the Rap proteins.

  17. [Biological Function of The Small G Protein Rap].

    PubMed

    Li, Shan-Shan; Guo, Xiao-Xi; An, Shu; Yang, Yang; Liu, Ying; Xu, Tian-Rui

    2016-02-01

    Rap has different biological functions on intracellular signaling pathways, such as regulating cell polarity, cell proliferation, cell differentiation, cell adhesion and cell movement. Furthermore, at tissue and organ level, Rap controls the establishment of neural polarity, synaptic growth, synaptic plasticity, neuronal migration and so on. Rap belongs to Ras family which contains two subtypes, Rap1 and Rap2. By binding GTP or GDP Rap transform between active or inactive state, and plays an important role as a molecular switch. Moreover, in the signal pathway of tumor, Rap inhibits cell transformation induced by the oncogene Ras, therefore inhibits the proliferation, invasion and migration of certain cancer cells by interacting with its downstream target molecules. In this review, we summarized the biological functions of Rap and discussed It's significance in cancer therapy and drug treatment of neurological diseases.

  18. [Biological Function of The Small G Protein Rap].

    PubMed

    Li, Shan-Shan; Guo, Xiao-Xi; An, Shu; Yang, Yang; Liu, Ying; Xu, Tian-Rui

    2016-02-01

    Rap has different biological functions on intracellular signaling pathways, such as regulating cell polarity, cell proliferation, cell differentiation, cell adhesion and cell movement. Furthermore, at tissue and organ level, Rap controls the establishment of neural polarity, synaptic growth, synaptic plasticity, neuronal migration and so on. Rap belongs to Ras family which contains two subtypes, Rap1 and Rap2. By binding GTP or GDP Rap transform between active or inactive state, and plays an important role as a molecular switch. Moreover, in the signal pathway of tumor, Rap inhibits cell transformation induced by the oncogene Ras, therefore inhibits the proliferation, invasion and migration of certain cancer cells by interacting with its downstream target molecules. In this review, we summarized the biological functions of Rap and discussed It's significance in cancer therapy and drug treatment of neurological diseases. PMID:27424400

  19. Rap G protein signal in normal and disordered lymphohematopoiesis.

    PubMed

    Minato, Nagahiro

    2013-09-10

    Rap proteins (Rap1, Rap2a, b, c) are small molecular weight GTPases of the Ras family. Rap G proteins mediate diverse cellular events such as cell adhesion, proliferation, and gene activation through various signaling pathways. Activation of Rap signal is regulated tightly by several specific regulatory proteins including guanine nucleotide exchange factors and GTPase-activating proteins. Beyond cell biological studies, increasing attempts have been made in the past decade to define the roles of Rap signal in specific functions of normal tissue systems as well as in cancer. In the immune and hematopoietic systems, Rap signal plays crucial roles in the development and function of essentially all lineages of lymphocytes and hematopoietic cells, and importantly, deregulated Rap signal may lead to unique pathological conditions depending on the affected cell types, including various types of leukemia and autoimmunity. The phenotypical studies have unveiled novel, even unexpected functional aspects of Rap signal in cells from a variety of tissues, providing potentially important clues for controlling human diseases, including malignancy.

  20. Rap G protein signal in normal and disordered lymphohematopoiesis

    SciTech Connect

    Minato, Nagahiro

    2013-09-10

    Rap proteins (Rap1, Rap2a, b, c) are small molecular weight GTPases of the Ras family. Rap G proteins mediate diverse cellular events such as cell adhesion, proliferation, and gene activation through various signaling pathways. Activation of Rap signal is regulated tightly by several specific regulatory proteins including guanine nucleotide exchange factors and GTPase-activating proteins. Beyond cell biological studies, increasing attempts have been made in the past decade to define the roles of Rap signal in specific functions of normal tissue systems as well as in cancer. In the immune and hematopoietic systems, Rap signal plays crucial roles in the development and function of essentially all lineages of lymphocytes and hematopoietic cells, and importantly, deregulated Rap signal may lead to unique pathological conditions depending on the affected cell types, including various types of leukemia and autoimmunity. The phenotypical studies have unveiled novel, even unexpected functional aspects of Rap signal in cells from a variety of tissues, providing potentially important clues for controlling human diseases, including malignancy.

  1. Structural basis of Rap phosphatase inhibition by Phr peptides.

    PubMed

    Gallego del Sol, Francisca; Marina, Alberto

    2013-01-01

    Two-component systems, composed of a sensor histidine kinase and an effector response regulator (RR), are the main signal transduction devices in bacteria. In Bacillus, the Rap protein family modulates complex signaling processes mediated by two-component systems, such as competence, sporulation, or biofilm formation, by inhibiting the RR components involved in these pathways. Despite the high degree of sequence homology, Rap proteins exert their activity by two completely different mechanisms of action: inducing RR dephosphorylation or blocking RR binding to its target promoter. However the regulatory mechanism involving Rap proteins is even more complex since Rap activity is antagonized by specific signaling peptides (Phr) through a mechanism that remains unknown at the molecular level. Using X-ray analyses, we determined the structure of RapF, the anti-activator of competence RR ComA, alone and in complex with its regulatory peptide PhrF. The structural and functional data presented herein reveal that peptide PhrF blocks the RapF-ComA interaction through an allosteric mechanism. PhrF accommodates in the C-terminal tetratricopeptide repeat domain of RapF by inducing its constriction, a conformational change propagated by a pronounced rotation to the N-terminal ComA-binding domain. This movement partially disrupts the ComA binding site by triggering the ComA disassociation, whose interaction with RapF is also sterically impaired in the PhrF-induced conformation of RapF. Sequence analyses of the Rap proteins, guided by the RapF-PhrF structure, unveil the molecular basis of Phr recognition and discrimination, allowing us to relax the Phr specificity of RapF by a single residue change.

  2. Plakophilin 3 mediates Rap1-dependent desmosome assembly and adherens junction maturation

    PubMed Central

    Todorovic´, Viktor; Koetsier, Jennifer L.; Godsel, Lisa M.; Green, Kathleen J.

    2014-01-01

    The pathways driving desmosome and adherens junction assembly are temporally and spatially coordinated, but how they are functionally coupled is poorly understood. Here we show that the Armadillo protein plakophilin 3 (Pkp3) mediates both desmosome assembly and E-cadherin maturation through Rap1 GTPase, thus functioning in a manner distinct from the closely related plakophilin 2 (Pkp2). Whereas Pkp2 and Pkp3 share the ability to mediate the initial phase of desmoplakin (DP) accumulation at sites of cell–cell contact, they play distinct roles in later steps: Pkp3 is required for assembly of a cytoplasmic population of DP-enriched junction precursors, whereas Pkp2 is required for transfer of the precursors to the membrane. Moreover, Pkp3 forms a complex with Rap1 GTPase, promoting its activation and facilitating desmosome assembly. We show further that Pkp3 deficiency causes disruption of an E-cadherin/Rap1 complex required for adherens junction sealing. These findings reveal Pkp3 as a coordinator of desmosome and adherens junction assembly and maturation through its functional association with Rap1. PMID:25208567

  3. Creative Technology and Rap

    ERIC Educational Resources Information Center

    Ch'ien, Evelyn

    2011-01-01

    This paper describes how a linguistic form, rap, can evolve in tandem with technological advances and manifest human-machine creativity. Rather than assuming that the interplay between machines and technology makes humans robotic or machine-like, the paper explores how the pressure of executing artistic visions using technology can drive…

  4. Squaring to the Rap!

    ERIC Educational Resources Information Center

    Adams, Deborah

    2006-01-01

    This article describes an approach to teaching square dance that is advantageous for both the teacher and students. Lessons in dance become more meaningful to students when the music and vocabulary is consistent with experiences in their own lives. When students create their own squaring to the rap, lessons become more student-centered,…

  5. Infant Information Processing and Family History of Specific Language Impairment: Converging Evidence for RAP Deficits from Two Paradigms

    ERIC Educational Resources Information Center

    Choudhury, Naseem; Leppanen, Paavo H. T.; Leevers, Hilary J.; Benasich, April A.

    2007-01-01

    An infant's ability to process auditory signals presented in rapid succession (i.e. rapid auditory processing abilities [RAP]) has been shown to predict differences in language outcomes in toddlers and preschool children. Early deficits in RAP abilities may serve as a behavioral marker for language-based learning disabilities. The purpose of this…

  6. The transcription factor Rap1p is required for tolerance to cell-wall perturbing agents and for cell-wall maintenance in Saccharomyces cerevisiae.

    PubMed

    Azad, Gajendra Kumar; Singh, Vikash; Baranwal, Shivani; Thakare, Mayur Jankiram; Tomar, Raghuvir S

    2015-01-01

    Yeast repressor activator protein (Rap1p) is involved in genomic stability and transcriptional regulation. We explored the function of Rap1p in yeast physiology using Rap1p truncation mutants. Our results revealed that the N-terminal truncation of Rap1p (Rap1ΔN) leads to hypersensitivity towards elevated temperature and cell-wall perturbing agents. Cell wall analysis showed an increase in the chitin and glucan content in Rap1ΔN cells as compared with wild type cells. Accordingly, mutant cells had a twofold thicker cell wall, as observed by electron microscopy. Furthermore, Rap1ΔN cells had increased levels of phosphorylated Slt2p, a MAP kinase of the cell wall integrity pathway. Mutant cells also had elevated levels of cell wall integrity response transcripts. Taken together, our findings suggest a connection between Rap1p and cell wall homeostasis.

  7. A Health Newsletter To Teach Science Knowledge: BioRAP!

    ERIC Educational Resources Information Center

    Froman, Robin D.; Owen, Steven V.; Del Rio-Parent, Lourdes

    This research describes the evaluation of a science curriculum newsletter called BioRAP which serves as a vehicle to teach current health science content. The research objectives were to estimate the relationships of socioeconomic status, ethnic group, gender, grade, student ability, and classroom use characteristics with student knowledge and…

  8. Pull-down assay for analysis of integrin-mediated activation of Rap proteins in adherent platelets.

    PubMed

    Guidetti, Gianni Francesco; Torti, Mauro

    2014-01-01

    Rap1 GTPases operate as molecular switches by cycling between a GDP-bound inactive state and a GTP-bound active state and regulate several cellular pathways in response to different stimuli. Circulating blood platelets express high levels of Rap1 proteins, mainly Rap1b, which plays a critical role in platelet adhesion and activation. Rap1 is a key element in the inside-out signaling pathway leading to the conversion of integrins into the high-affinity state for their ligands. In platelets, Rap1b regulates inside-out activation of both integrin αIIbβ3 and α2β1. In addition, Rap1b is also involved in integrin outside-in signaling. Integrin-mediated platelet adhesion leads to accumulation of GTP-bound Rap1b, which promotes integrin-mediated processes such as spreading and clot retraction. Rap1b is thus a bidirectional regulator of platelet integrin function. Here we describe a method to analyze Rap1b activation induced by platelet adhesion via integrin α2β1.

  9. German Rap Music in the Classroom.

    ERIC Educational Resources Information Center

    Schmidt, Johannes

    2003-01-01

    Provides background information on German rap artists and bands and discusses how to implement the music in various classroom situations at all levels. Highlights some of the available material on German rap music and provides information on how to locate rap texts, information, and other material via the Internet and other sources. (Author/VWL)

  10. The Dorsal Rather than Ventral Pathway Better Reflects Individual Syntactic Abilities in Second Language

    PubMed Central

    Yamamoto, Kayako; Sakai, Kuniyoshi L.

    2016-01-01

    The left inferior frontal gyrus (IFG) has been reported to be critically involved in syntactic processing, not only in first language (L1), but in second language (L2). Indeed, the leftward lateralization of the IFG has been shown to be correlated with the performance of a syntactic task in L2. Given that posterior language-related regions are systematically connected with the left IFG, the next question is which of the dorsal and ventral pathways is more critical to the individual syntactic abilities in L2. Here we used diffusion magnetic resonance imaging (MRI) and tractography with newly developed semi-automatic methods of defining seeds and selecting regions of interest (ROIs). We calculated mean thickness and fractional anisotropy (FA) in each ROI for the arcuate fasciculus (Arcuate) of the dorsal pathway, as well as for the inferior fronto-occipital fasciculus (IFOF) of the ventral pathway. In Experiment I, we performed partial correlation analyses between FA and the accuracy of the syntactic task, removing the effects of the accuracy of a spelling task, gender, and handedness. Among the two pathways in each hemisphere, only FA of the left Arcuate was significantly correlated with individual accuracy of the syntactic task. In Experiment II, we recruited monozygotic twins and examined to what extent their L2 abilities and their structural properties were similar. Within twin pairs, the highest significant correlation was observed for reaction times of the spelling task, while the correlation for the accuracy of the syntactic task was marginal; these two correlation coefficients were significantly different. Moreover, the thickness of the left Arcuate was highly correlated within pairs, while its FA, as well as the thickness/FA in the ventral pathways, was not significantly correlated. The correlation coefficient for the thickness of the left Arcuate was significantly larger than that of the left IFOF. These results suggest that the thickness of the left

  11. The Dorsal Rather than Ventral Pathway Better Reflects Individual Syntactic Abilities in Second Language.

    PubMed

    Yamamoto, Kayako; Sakai, Kuniyoshi L

    2016-01-01

    The left inferior frontal gyrus (IFG) has been reported to be critically involved in syntactic processing, not only in first language (L1), but in second language (L2). Indeed, the leftward lateralization of the IFG has been shown to be correlated with the performance of a syntactic task in L2. Given that posterior language-related regions are systematically connected with the left IFG, the next question is which of the dorsal and ventral pathways is more critical to the individual syntactic abilities in L2. Here we used diffusion magnetic resonance imaging (MRI) and tractography with newly developed semi-automatic methods of defining seeds and selecting regions of interest (ROIs). We calculated mean thickness and fractional anisotropy (FA) in each ROI for the arcuate fasciculus (Arcuate) of the dorsal pathway, as well as for the inferior fronto-occipital fasciculus (IFOF) of the ventral pathway. In Experiment I, we performed partial correlation analyses between FA and the accuracy of the syntactic task, removing the effects of the accuracy of a spelling task, gender, and handedness. Among the two pathways in each hemisphere, only FA of the left Arcuate was significantly correlated with individual accuracy of the syntactic task. In Experiment II, we recruited monozygotic twins and examined to what extent their L2 abilities and their structural properties were similar. Within twin pairs, the highest significant correlation was observed for reaction times of the spelling task, while the correlation for the accuracy of the syntactic task was marginal; these two correlation coefficients were significantly different. Moreover, the thickness of the left Arcuate was highly correlated within pairs, while its FA, as well as the thickness/FA in the ventral pathways, was not significantly correlated. The correlation coefficient for the thickness of the left Arcuate was significantly larger than that of the left IFOF. These results suggest that the thickness of the left

  12. Rap Music: An Education with a Beat from the Street.

    ERIC Educational Resources Information Center

    Powell, Catherine Tabb

    1991-01-01

    Examines rap music's origins as street music and poetry in the early 1970s and as an expression of Black youth experience. Discusses rap groups, women and Whites in rap, Christian rap, copyright disputes, and distribution difficulties. Rap is an informal educational medium that affects adolescents' values and attitudes. (JB)

  13. Rap-Music Attitude and Perception Scale: A Validation Study

    ERIC Educational Resources Information Center

    Tyson, Edgar H.

    2006-01-01

    Objective: This study tests the validity of the Rap-music Attitude and Perception (RAP) Scale, a 1-page, 24-item measure of a person's thoughts and feelings surrounding the effects and content of rap music. The RAP was designed as a rapid assessment instrument for youth programs and practitioners using rap music and hip hop culture in their work…

  14. Regulation of Rap GTPases in mammalian neurons.

    PubMed

    Shah, Bhavin; Püschel, Andreas W

    2016-10-01

    Small GTPases are central regulators of many cellular processes. The highly conserved Rap GTPases perform essential functions in the mammalian nervous system during development and in mature neurons. During neocortical development, Rap1 is required to regulate cadherin- and integrin-mediated adhesion. In the adult nervous system Rap1 and Rap2 regulate the maturation and plasticity of dendritic spine and synapses. Although genetic studies have revealed important roles of Rap GTPases in neurons, their regulation by guanine nucleotide exchange factors (GEFs) that activate them and GTPase activating proteins (GAPs) that inactivate them by stimulating their intrinsic GTPase activity is just beginning to be explored in vivo. Here we review how GEFs and GAPs regulate Rap GTPases in the nervous system with a focus on their in vivo function.

  15. Regulation of Rap GTPases in mammalian neurons.

    PubMed

    Shah, Bhavin; Püschel, Andreas W

    2016-10-01

    Small GTPases are central regulators of many cellular processes. The highly conserved Rap GTPases perform essential functions in the mammalian nervous system during development and in mature neurons. During neocortical development, Rap1 is required to regulate cadherin- and integrin-mediated adhesion. In the adult nervous system Rap1 and Rap2 regulate the maturation and plasticity of dendritic spine and synapses. Although genetic studies have revealed important roles of Rap GTPases in neurons, their regulation by guanine nucleotide exchange factors (GEFs) that activate them and GTPase activating proteins (GAPs) that inactivate them by stimulating their intrinsic GTPase activity is just beginning to be explored in vivo. Here we review how GEFs and GAPs regulate Rap GTPases in the nervous system with a focus on their in vivo function. PMID:27186679

  16. Rap Music in School Counseling Based on Don Elligan's Rap Therapy

    ERIC Educational Resources Information Center

    Gonzalez, Tiphanie; Hayes, B. Grant

    2009-01-01

    In 2000, Don Elligan introduced Rap Therapy as a psychotherapeutic intervention for working with at-risk youths, primarily African American males whose identities were highly influenced by rap music. Rap music can engage a population of youth who often enter counseling apprehensively (Elligan 2000, 2004; Tillie-Allen, 2005). This article reviews…

  17. TRF2-RAP1 is required to protect telomeres from engaging in homologous recombination-mediated deletions and fusions.

    PubMed

    Rai, Rekha; Chen, Yong; Lei, Ming; Chang, Sandy

    2016-01-01

    Repressor/activator protein 1 (RAP1) is a highly conserved telomere-interacting protein. Yeast Rap1 protects telomeres from non-homologous end joining (NHEJ), plays important roles in telomere length control and is involved in transcriptional gene regulation. However, a role for mammalian RAP1 in telomere end protection remains controversial. Here we present evidence that mammalian RAP1 is essential to protect telomere from homology directed repair (HDR) of telomeres. RAP1 cooperates with the basic domain of TRF2 (TRF2(B)) to repress PARP1 and SLX4 localization to telomeres. Without RAP1 and TRF2(B), PARP1 and SLX4 HR factors promote rapid telomere resection, resulting in catastrophic telomere loss and the generation of telomere-free chromosome fusions in both mouse and human cells. The RAP1 Myb domain is required to repress both telomere loss and formation of telomere-free fusions. Our results highlight the importance of the RAP1-TRF2 heterodimer in protecting telomeres from inappropriate processing by the HDR pathway. PMID:26941064

  18. Roles of JnRAP2.6-like from the Transition Zone of Black Walnut in Hormone Signaling

    PubMed Central

    Huang, Zhonglian; Zhao, Peng; Medina, Jose; Meilan, Richard; Woeste, Keith

    2013-01-01

    An EST sequence, designated JnRAP2-like, was isolated from tissue at the heartwood/sapwood transition zone (TZ) in black walnut (Juglans nigra L). The deduced amino acid sequence of JnRAP2-like protein consists of a single AP2-containing domain with significant similarity to conserved AP2/ERF DNA-binding domains in other species. Based on multiple sequence alignment, JnRAP2-like appears to be an ortholog of RAP2.6L (At5g13330), which encodes an ethylene response element binding protein in Arabidopsis thaliana. Real-time PCR revealed that the JnRAP2-like was expressed most abundantly in TZ of trees harvested in fall when compared with other xylem tissues harvested in the fall or summer. Independent transgenic lines over-expressing JnRAP2-like in Arabidopsis developed dramatic ethylene-related phenotypes when treated with 50 µM methyl jasmonate (MeJA). Taken together, these results indicated that JnRAP2-like may participate in the integration of ethylene and jasmonate signals in the xylem and other tissues. Given the role of ethylene in heartwood formation, it is possible JnRAP2-like expression in the transition zone is part of the signal transduction pathway leading to heartwood formation in black walnut. PMID:24265672

  19. Overexpression of Rap-1A indicates a poor prognosis for oral cavity squamous cell carcinoma and promotes tumor cell invasion via Aurora-A modulation.

    PubMed

    Chen, Chang-Han; Chuang, Hui-Ching; Huang, Chao-Cheng; Fang, Fu-Min; Huang, Hsuan-Ying; Tsai, Hsin-Ting; Su, Li-Jen; Shiu, Li-Yen; Leu, Steve; Chien, Chih-Yen

    2013-02-01

    The functions of Rap-1A in oral carcinogenesis are largely unexplored. In this study, we examined the expression of Rap-1A at different malignant stages of oral cavity squamous cell carcinoma (OCSCC). Semiquantitative RT-PCR, quantitative RT-PCR, and Western blotting were used to evaluate Rap-1A mRNA and protein expressions, respectively, in paired OCSCC patient specimens. To determine the possible correlation between Rap-1A expression and various clinical characteristics, 256 samples from patients with OCSCC were evaluated by immunohistochemical staining. Strong Rap-1A expression was a significant prognostic marker and predictor of aggressive OCSCC. The overall and disease-specific 5-year survival rates were significantly correlated with strong expression of Rap-1A (P < 0.001). Functionally, overexpressed Rap-1A could promote oral cancer cell migration and invasion by Transwell chambers and wound healing assay. Conversely, the suppression of Rap-1A expression using Rap-1A-mediated siRNA was sufficient to decrease cell motility. Furthermore, our data also illustrated that Aurora-A could not only induce mRNA and protein expressions of Rap-1A for enhancing cancer cell motility but also co-localize and form a complex with Rap-1A in the oral cancer cell line. Finally, immunohistochemical staining, indirect immunofluorescence, and Western blotting analysis of human aggressive OCSCC specimens revealed a significantly positive correlation between Rap-1A and Aurora-A expression. Taken together, our results suggest that the Aurora-A/Rap-1A pathway is associated with survival, tumor progression, and metastasis of OCSCC patients.

  20. Hippocampal Pathway Plasticity Is Associated with the Ability to Form Novel Memories in Older Adults

    PubMed Central

    Antonenko, Daria; Külzow, Nadine; Cesarz, Magda E.; Schindler, Kristina; Grittner, Ulrike; Flöel, Agnes

    2016-01-01

    White matter deterioration in the aging human brain contributes to cognitive decline. The fornix as main efferent hippocampal pathway is one of the tracts most strongly associated with age-related memory impairment. Its deterioration may predict conversion to Alzheimer’s dementia and its precursors. However, the associations between the ability to form novel memories, fornix microstructure and plasticity in response to training have never been tested. In the present study, 25 healthy older adults (15 women; mean age (SD): 69 (6) years) underwent an object-location training on three consecutive days. Behavioral outcome measures comprised recall performance on the training days, and on 1-day and 1-month follow up assessments. MRI at 3 Tesla was assessed before and after training. Fornix microstructure was determined by fractional anisotropy and mean diffusivity (MD) values from diffusion tensor imaging (DTI). In addition, hippocampal volumes were extracted from high-resolution images; individual hippocampal masks were further aligned to DTI images to determine hippocampal microstructure. Using linear mixed model analysis, we found that the change in fornix FA from pre- to post-training assessment was significantly associated with training success. Neither baseline fornix microstructure nor hippocampal microstructure or volume changes were significantly associated with performance. Further, models including control task performance (auditory verbal learning) and control white matter tract microstructure (uncinate fasciculus and parahippocampal cingulum) did not yield significant associations. Our results confirm that hippocampal pathways respond to short-term cognitive training, and extend previous findings by demonstrating that the magnitude of training-induced structural changes is associated with behavioral success in older adults. This suggests that the amount of fornix plasticity may not only be behaviorally relevant, but also a potential sensitive biomarker

  1. E-cadherin dis-engagement activates the Rap1 GTPase

    PubMed Central

    Asuri, Sirisha; Yan, Jingliang; Paranavitana, Nivanka C.; Quilliam, Lawrence A.

    2008-01-01

    E-cadherin based adherens junctions are finely regulated by multiple cellular signaling events. Here we show that the Ras-related Rap1 GTPase is enriched in regions of nascent cell-cell contacts and strengthens E-cadherin junctions: constitutively active Rap1 expressing MDCK cells exhibit increased junctional contact and resisted calcium depletion-induced cell-cell junction disruption. E-cadherin disengagement activated Rap1 and this correlated with E-cadherin association with the Rap GEFs, C3G and PDZ-GEF I. PDZ-GEF I associated with E-cadherin and β-catenin whereas C3G interaction with E-cadherin did not involve β-catenin. Knockdown of PDZ-GEF I in MDCK cells decreased Rap1 activity following E-cadherin junction disruption. We hereby show that Rap1 plays a role in the maintenance and repair of E-cadherin junctions and is activated via an “outside-in” signaling pathway initiated by E-cadherin and mediated at least in part by PDZ-GEF I. PMID:18767072

  2. Unwrapping Rap: A Literacy of Lived Experience.

    ERIC Educational Resources Information Center

    Brown, Stephen G.

    The adversarial forces of governmental censorship, freedom of expression, and capitalistic appropriation are engaged in an acrimonious debate over "Gangsta' Rap" that is being played out in the public spaces of popular culture. However, as a literacy of lived experience, Gangsta' Rap warrants critical investigation. Many postmodern theorists have…

  3. Black Essentialism: The Art of Jazz Rap.

    ERIC Educational Resources Information Center

    Stewart, Earl; Duran, Jane

    1999-01-01

    Establishes a black essentialist aesthetic for jazz rap, showing its relation to an African-derived history and other black traditions. Examines newer lines of argument in aesthetics about contemporary recordings focusing on Theodore Gracyk. Argues that jazz rap is defined by actual, not recorded, performance. (CMK)

  4. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner or... through 270.66 do not apply to RAPs unless those requirements for traditional RCRA permits are... requirements in this section constitutes a RCRA permit under RCRA section 3005(c). (d) A RAP may be: (1)...

  5. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner or... through 270.66 do not apply to RAPs unless those requirements for traditional RCRA permits are... requirements in this section constitutes a RCRA permit under RCRA section 3005(c). (d) A RAP may be: (1)...

  6. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner or... through 270.66 do not apply to RAPs unless those requirements for traditional RCRA permits are... requirements in this section constitutes a RCRA permit under RCRA section 3005(c). (d) A RAP may be: (1)...

  7. 40 CFR 270.100 - Who must obtain a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false Who must obtain a RAP? 270.100 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.100 Who must obtain a RAP? When a facility or remediation waste management site...

  8. 40 CFR 270.100 - Who must obtain a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Who must obtain a RAP? 270.100 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.100 Who must obtain a RAP? When a facility or remediation waste management site...

  9. 40 CFR 270.100 - Who must obtain a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false Who must obtain a RAP? 270.100 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.100 Who must obtain a RAP? When a facility or remediation waste management site...

  10. 40 CFR 270.195 - When will my RAP expire?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false When will my RAP expire? 270.195... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.195 When will my RAP...

  11. 40 CFR 270.195 - When will my RAP expire?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false When will my RAP expire? 270.195... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.195 When will my RAP...

  12. 40 CFR 270.100 - Who must obtain a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false Who must obtain a RAP? 270.100 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.100 Who must obtain a RAP? When a facility or remediation waste management site...

  13. 40 CFR 270.100 - Who must obtain a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false Who must obtain a RAP? 270.100 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.100 Who must obtain a RAP? When a facility or remediation waste management site...

  14. 40 CFR 270.195 - When will my RAP expire?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false When will my RAP expire? 270.195... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.195 When will my RAP...

  15. 40 CFR 270.195 - When will my RAP expire?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false When will my RAP expire? 270.195... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.195 When will my RAP...

  16. 40 CFR 270.195 - When will my RAP expire?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false When will my RAP expire? 270.195... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.195 When will my RAP...

  17. How Does Academic Ability Affect Educational and Labour Market Pathways in Canada. OECD Education Working Papers, No. 30

    ERIC Educational Resources Information Center

    Hansen, Jorgen

    2010-01-01

    Using data from the Youth in Transition Survey (YITS), this paper provides an up-to-date description of educational and labour market pathways (or transitions) among Canadian youth. It also estimates the effect of academic abilities, measured by PISA math and reading scores, on such transitions. Descriptive statistics show that educational success…

  18. Rho family and Rap GTPase activation assays.

    PubMed

    Jennings, Richard T; Knaus, Ulla G

    2014-01-01

    The detection of Ras superfamily GTPase activity in innate immune cells is important when studying signaling events elicited by various ligands and cellular processes. The development of high-affinity probes detecting the activated, GTP-bound form of small GTPases has significantly enhanced our understanding of initiation and termination of GTPase-regulated signaling pathways. These probes are created by fusing a high-affinity GTPase-binding domain derived from a specific downstream effector protein to glutathione S-transferase (GST). Such domains bind preferentially to the GTP-bound form of the upstream Rho or Ras GTPase. Coupling these probes to beads enables extraction of the complex and subsequent quantification of the active GTP-binding protein by immunoblotting. Although effector domains that discriminate efficiently between GDP- and GTP-bound states and highly specific antibodies are not yet available for every small GTPase, analysis of certain members of the Rho and Ras GTPase family is now routinely performed. Here, we describe affinity-based pulldown assays for detection of Rho GTPase (Rac1/2, Cdc42, RhoA/B) and Rap1/2 activity in stimulated neutrophils or macrophages.

  19. Neuronal Rap1 regulates energy balance, glucose homeostasis, and leptin actions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Central Nervous System (CNS) contributes to obesity and metabolic disease; however, the underlying neurobiological pathways remain to be fully established. Here, we show that the small GTPase Rap1 is expressed in multiple hypothalamic nuclei that control whole-body metabolism and is activated in...

  20. A Rap GTPase interactor, RADIL, mediates migration of neural crest precursors.

    PubMed

    Smolen, Gromoslaw A; Schott, Benjamin J; Stewart, Rodney A; Diederichs, Sven; Muir, Beth; Provencher, Heather L; Look, A Thomas; Sgroi, Dennis C; Peterson, Randall T; Haber, Daniel A

    2007-09-01

    The neural crest (NC) is a highly motile cell population that gives rise to multiple tissue lineages during vertebrate embryogenesis. Here, we identify a novel effector of the small GTPase Rap, called RADIL, and show that it is required for cell adhesion and migration. Knockdown of radil in the zebrafish model results in multiple defects in NC-derived lineages such as cartilage, pigment cells, and enteric neurons. We specifically show that these defects are primarily due to the diminished migratory capacity of NC cells. The identification of RADIL as a regulator of NC migration defines a role for the Rap pathway in this process.

  1. Molecular characterization of the Babesia caballi rap-1 gene and epidemiological survey in horses in Israel.

    PubMed

    Rapoport, Adi; Aharonson-Raz, Karin; Berlin, Dalia; Tal, Saar; Gottlieb, Yuval; Klement, Eyal; Steinman, Amir

    2014-04-01

    Equine piroplasmosis imposes great concerns for the equine industry regarding international horse movement, and therefore requires reliable diagnostic tools. Recent studies from South Africa and Jordan, including a preliminary study in Israel, reported extremely low seroprevalence to Babesia caballi (B. caballi) (0-1%) using the acceptable rhoptry-associated protein-1 (RAP-1) cELISA. In accordance with the study from South Africa demonstrating a significant heterogeneity in the rap-1 gene sequence of South African B. caballi isolates, the objectives of this study were to phylogenetically characterize the rap-1 gene of the Israeli isolates and determine the prevalence of B. caballi in horses in Israel. Out of 273 horses tested using the RAP-1 cELISA, only one was sero-positive, while 9.3% were positive on PCR performed on the rap-1 gene. Phylogenetic analysis of the rap-1 gene grouped the Israeli isolates in a cluster together with the South African strains (99% nt identity), but in a separate cluster from the American/Caribbean strains (81-82% nt identity). These findings support the existence of heterogeneity in the RAP-1 amino-acid sequences of the Israeli and South African isolates as compared to that used in the cELISA commercial kit and raise doubts as to the ability of this assay to serve as a sole regulatory test for international horse movement. Risk factor analysis found management and age to significantly associate with prevalence of B. caballi, as higher prevalence was noted in horses held out on pasture and a negative association was recorded with age. In addition, B. caballi was not detected in horses in the steppe-arid and extreme-arid climatic regions as compared to the wetter regions. Findings of this study emphasize the need to combine several detection methods to ameliorate the control and spread of the disease.

  2. Neuronal Rap1 Regulates Energy Balance, Glucose Homeostasis, and Leptin Actions.

    PubMed

    Kaneko, Kentaro; Xu, Pingwen; Cordonier, Elizabeth L; Chen, Siyu S; Ng, Amy; Xu, Yong; Morozov, Alexei; Fukuda, Makoto

    2016-09-13

    The CNS contributes to obesity and metabolic disease; however, the underlying neurobiological pathways remain to be fully established. Here, we show that the small GTPase Rap1 is expressed in multiple hypothalamic nuclei that control whole-body metabolism and is activated in high-fat diet (HFD)-induced obesity. Genetic ablation of CNS Rap1 protects mice from dietary obesity, glucose imbalance, and insulin resistance in the periphery and from HFD-induced neuropathological changes in the hypothalamus, including diminished cellular leptin sensitivity and increased endoplasmic reticulum (ER) stress and inflammation. Furthermore, pharmacological inhibition of CNS Rap1 signaling normalizes hypothalamic ER stress and inflammation, improves cellular leptin sensitivity, and reduces body weight in mice with dietary obesity. We also demonstrate that Rap1 mediates leptin resistance via interplay with ER stress. Thus, neuronal Rap1 critically regulates leptin sensitivity and mediates HFD-induced obesity and hypothalamic pathology and may represent a potential therapeutic target for obesity treatment. PMID:27626668

  3. RASA3 is a critical inhibitor of RAP1-dependent platelet activation

    PubMed Central

    Stefanini, Lucia; Paul, David S.; Robledo, Raymond F.; Chan, E. Ricky; Getz, Todd M.; Campbell, Robert A.; Kechele, Daniel O.; Casari, Caterina; Piatt, Raymond; Caron, Kathleen M.; Mackman, Nigel; Weyrich, Andrew S.; Parrott, Matthew C.; Boulaftali, Yacine; Adams, Mark D.; Peters, Luanne L.; Bergmeier, Wolfgang

    2015-01-01

    The small GTPase RAP1 is critical for platelet activation and thrombus formation. RAP1 activity in platelets is controlled by the GEF CalDAG-GEFI and an unknown regulator that operates downstream of the adenosine diphosphate (ADP) receptor, P2Y12, a target of antithrombotic therapy. Here, we provide evidence that the GAP, RASA3, inhibits platelet activation and provides a link between P2Y12 and activation of the RAP1 signaling pathway. In mice, reduced expression of RASA3 led to premature platelet activation and markedly reduced the life span of circulating platelets. The increased platelet turnover and the resulting thrombocytopenia were reversed by concomitant deletion of the gene encoding CalDAG-GEFI. Rasa3 mutant platelets were hyperresponsive to agonist stimulation, both in vitro and in vivo. Moreover, activation of Rasa3 mutant platelets occurred independently of ADP feedback signaling and was insensitive to inhibitors of P2Y12 or PI3 kinase. Together, our results indicate that RASA3 ensures that circulating platelets remain quiescent by restraining CalDAG-GEFI/RAP1 signaling and suggest that P2Y12 signaling is required to inhibit RASA3 and enable sustained RAP1-dependent platelet activation and thrombus formation at sites of vascular injury. These findings provide insight into the antithrombotic effect of P2Y12 inhibitors and may lead to improved diagnosis and treatment of platelet-related disorders. PMID:25705885

  4. Hypoxia/reoxygenation-experienced cancer cell migration and metastasis are regulated by Rap1- and Rac1-GTPase activation via the expression of thymosin beta-4.

    PubMed

    Lee, Jae-Wook; Ryu, Yun-Kyoung; Ji, Young-Hoon; Kang, Joo Hyun; Moon, Eun-Yi

    2015-01-01

    Signaling by small guanosine triphosphatases (GTPase), Rap1/Rac1, is one of the major pathways controlling cancer cell migration and tumor metastasis. Thymosin beta-4 (Tβ4), an actin-sequestering protein, has been shown to increase migration of cancer cells. Episodes of hypoxia and re-oxygenation (H/R) are an important phenomenon in tumor microenvironment (TME). We investigated whether Tβ4 could play as an intermediary to crosstalk between Rac1- and Rap1- GTPase activation under hypoxia/reoxygenation (H/R) conditions. Inhibition of Tβ4 expression using transcription activator-like effector nucleases (TALEN) significantly decreased lung metastasis of B16F10 cells. Rac1 and Rap1 activity, as well as cancer cell migration, increased following induction of Tβ4 expression in normoxia- or H/R-experienced cells, but were barely detectable in Tβ4-depleted cells. Rap1-regulated Rac1 activity was decreased by a dominant negative Rap1 (Rap1N17), and increased by 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate (CPT), a Rap1 activator. In contrast, a Rac1-specific inhibitor, NSC23766, and dominant negative Rac1 (Rac1N17) enhanced Tβ4 expression and aberrant Rap1 activity. While NSC23766 and Rac1N17 incompletely inhibited tumor metastasis in vivo, and H/R-experienced cancer cell migration in vitro, more efficient attenuation of cancer cell migration was accomplished by simultaneous inactivation of Rap1 and Rac1 with Rap1N17 and Rac1N17, respectively. These data suggest that a combination therapy targeting both Rap1 and Rac1 activity may be an effective method of inhibiting tumor metastasis.

  5. The Rap-RapGAP complex: GTP hydrolysis without catalytic glutamine and arginine residues.

    PubMed

    Scrima, Andrea; Thomas, Christoph; Deaconescu, Delia; Wittinghofer, Alfred

    2008-04-01

    The GTP-binding protein Rap1 regulates integrin-mediated and other cell adhesion processes. Unlike most other Ras-related proteins, it contains a threonine in switch II instead of a glutamine (Gln61 in Ras), a residue crucial for the GTPase reaction of most G proteins. Furthermore, unlike most other GTPase-activating proteins (GAPs) for small G proteins, which supply a catalytically important Arg-finger, no arginine residue of RapGAP makes a significant contribution to the GTPase reaction of Rap1. For a detailed understanding of the reaction mechanism, we have solved the structure of Rap1 in complex with Rap1GAP. It shows that the Thr61 of Rap is away from the active site and that an invariant asparagine of RapGAPs, the Asn-thumb, takes over the role of the cis-glutamine of Ras, Rho or Ran. The structure and biochemical data allow to further explain the mechanism and to define the important role of a conserved tyrosine. The structure and biochemical data furthermore show that the RapGAP homologous region of the tumour suppressor Tuberin is sufficient for catalysis on Rheb.

  6. Activation of JNK by Epac is independent of its activity as a Rap guanine nucleotide exchanger.

    PubMed

    Hochbaum, Daniel; Tanos, Tamara; Ribeiro-Neto, Fernando; Altschuler, Daniel; Coso, Omar A

    2003-09-01

    Guanine nucleotide exchange factors (GEFs) and their associated GTP-binding proteins (G-proteins) are key regulatory elements in the signal transduction machinery that relays information from the extracellular environment into specific intracellular responses. Among them, the MAPK cascades represent ubiquitous downstream effector pathways. We have previously described that, analogous to the Ras-dependent activation of the Erk-1/2 pathway, members of the Rho family of small G-proteins activate the JNK cascade when GTP is loaded by their corresponding GEFs. Searching for novel regulators of JNK activity we have identified Epac (exchange protein activated by cAMP) as a strong activator of JNK-1. Epac is a member of a growing family of GEFs that specifically display exchange activity on the Rap subfamily of Ras small G-proteins. We report here that while Epac activates the JNK severalfold, a constitutively active (G12V) mutant of Rap1b does not, suggesting that Rap-GTP is not sufficient to transduce Epac-dependent JNK activation. Moreover, Epac signaling to the JNKs was not blocked by inactivation of endogenous Rap, suggesting that Rap activation is not necessary for this response. Consistent with these observations, domain deletion mutant analysis shows that the catalytic GEF domain is dispensable for Epac-mediated activation of JNK. These studies identified a region overlapping the Ras exchange motif domain as critical for JNK activation. Consistent with this, an isolated Ras exchange motif domain from Epac is sufficient to activate JNK. We conclude that Epac signals to the JNK cascade through a new mechanism that does not involve its canonical catalytic action, i.e. Rap-specific GDP/GTP exchange. This represents not only a novel way to activate the JNKs but also a yet undescribed mechanism of downstream signaling by Epac.

  7. Change of function of the wheat stress-responsive transcriptional repressor TaRAP2.1L by repressor motif modification.

    PubMed

    Amalraj, Amritha; Luang, Sukanya; Kumar, Manoj Yadav; Sornaraj, Pradeep; Eini, Omid; Kovalchuk, Nataliya; Bazanova, Natalia; Li, Yuan; Yang, Nannan; Eliby, Serik; Langridge, Peter; Hrmova, Maria; Lopato, Sergiy

    2016-02-01

    Plants respond to abiotic stresses by changes in gene regulation, including stress-inducible expression of transcriptional activators and repressors. One of the best characterized families of drought-related transcription factors are dehydration-responsive element binding (DREB) proteins, known as C-repeat binding factors (CBF). The wheat DREB/CBF gene TaRAP2.1L was isolated from drought-affected tissues using a dehydration-responsive element (DRE) as bait in a yeast one-hybrid screen. TaRAP2.1L is induced by elevated abscisic acid, drought and cold. A C-terminal ethylene responsive factor-associated amphiphilic repression (EAR) motif, known to be responsible for active repression of target genes, was identified in the TaRAP2.1L protein. It was found that TaRAP2.1L has a unique selectivity of DNA-binding, which differs from that of DREB activators. This binding selectivity remains unchanged in a TaRAP2.1L variant with an inactivated EAR motif (TaRAP2.1Lmut). To study the role of the TaRAP2.1L repressor activity associated with the EAR motif in planta, transgenic wheat overexpressing native or mutated TaRAP2.1L was generated. Overexpression of TaRAP2.1L under constitutive and stress-inducible promoters in transgenic wheat and barley led to dwarfism and decreased frost tolerance. By contrast, constitutive overexpression of the TaRAP2.1Lmut gene had little or no negative influence on wheat development or grain yield. Transgenic lines with the TaRAP2.1Lmut transgene had an enhanced ability to survive frost and drought. The improved stress tolerance is attributed to up-regulation of several stress-related genes known to be downstream genes of DREB/CBF activators.

  8. The NS1 Protein of Influenza A Virus Interacts with Cellular Processing Bodies and Stress Granules through RNA-Associated Protein 55 (RAP55) during Virus Infection

    PubMed Central

    Mok, Bobo Wing-Yee; Song, Wenjun; Wang, Pui; Tai, Hung; Chen, Yixin; Zheng, Min; Wen, Xi; Lau, Siu-Ying; Wu, Wai Lan; Matsumoto, Ken

    2012-01-01

    The nonstructural protein (NS1) of influenza A virus performs multiple functions in the virus life cycle. Proteomic screening for cellular proteins which interact with NS1 identified the cellular protein RAP55, which is one of the components of cellular processing bodies (P-bodies) and stress granules. To verify whether NS1 interacts with cellular P-bodies, interactions between NS1, RAP55, and other P-body-associated proteins (Ago1, Ago2, and DCP1a) were confirmed using coimmunoprecipitation and cellular colocalization assays. Overexpression of RAP55 induced RAP55-associated stress granule formation and suppressed virus replication. Knockdown of RAP55 with small interfering RNA (siRNA) or expression of a dominant-negative mutant RAP55 protein with defective interaction with P-bodies blocked NS1 colocalization to P-bodies in cells. Expression of NS1 inhibited RAP55 expression and formation of RAP55-associated P-bodies/stress granules. The viral nucleoprotein (NP) was found to be targeted to stress granules in the absence of NS1 but localized to P-bodies when NS1 was coexpressed. Restriction of virus replication via P-bodies occurred in the early phases of infection, as the number of RAP55-associated P-bodies in cells diminished over the course of virus infection. NS1 interaction with RAP55-associated P-bodies/stress granules was associated with RNA binding and mediated via a protein kinase R (PKR)-interacting viral element. Mutations introduced into either RNA binding sites (R38 and K41) or PKR interaction sites (I123, M124, K126, and N127) caused NS1 proteins to lose the ability to interact with RAP55 and to inhibit stress granules. These results reveal an interplay between virus and host during virus replication in which NP is targeted to P-bodies/stress granules while NS1 counteracts this host restriction mechanism. PMID:22973032

  9. Rap Music and Its Violent Progeny: America's Culture of Violence in Context.

    ERIC Educational Resources Information Center

    Richardson, Jeanita W.; Scott, Kim A.

    2002-01-01

    Considers rap music as a creative expression and metaphorical offspring of America's well-established culture of violence, highlighting rap music in the context of a violent culture; violence in music; rap, cultural capital, and social reproduction; rap in the scholarly literature; political and judicial scrutiny of rap; and capitalism and rap.…

  10. Cdk5-mediated phosphorylation of RapGEF2 controls neuronal migration in the developing cerebral cortex.

    PubMed

    Ye, Tao; Ip, Jacque P K; Fu, Amy K Y; Ip, Nancy Y

    2014-01-01

    During cerebral cortex development, pyramidal neurons migrate through the intermediate zone and integrate into the cortical plate. These neurons undergo the multipolar-bipolar transition to initiate radial migration. While perturbation of this polarity acquisition leads to cortical malformations, how this process is initiated and regulated is largely unknown. Here we report that the specific upregulation of the Rap1 guanine nucleotide exchange factor, RapGEF2, in migrating neurons corresponds to the timing of this polarity transition. In utero electroporation and live-imaging studies reveal that RapGEF2 acts on the multipolar-bipolar transition during neuronal migration via a Rap1/N-cadherin pathway. Importantly, activation of RapGEF2 is controlled via phosphorylation by a serine/threonine kinase Cdk5, whose activity is largely restricted to the radial migration zone. Thus, the specific expression and Cdk5-dependent phosphorylation of RapGEF2 during multipolar-bipolar transition within the intermediate zone are essential for proper neuronal migration and wiring of the cerebral cortex. PMID:25189171

  11. Cdk5-mediated phosphorylation of RapGEF2 controls neuronal migration in the developing cerebral cortex.

    PubMed

    Ye, Tao; Ip, Jacque P K; Fu, Amy K Y; Ip, Nancy Y

    2014-01-01

    During cerebral cortex development, pyramidal neurons migrate through the intermediate zone and integrate into the cortical plate. These neurons undergo the multipolar-bipolar transition to initiate radial migration. While perturbation of this polarity acquisition leads to cortical malformations, how this process is initiated and regulated is largely unknown. Here we report that the specific upregulation of the Rap1 guanine nucleotide exchange factor, RapGEF2, in migrating neurons corresponds to the timing of this polarity transition. In utero electroporation and live-imaging studies reveal that RapGEF2 acts on the multipolar-bipolar transition during neuronal migration via a Rap1/N-cadherin pathway. Importantly, activation of RapGEF2 is controlled via phosphorylation by a serine/threonine kinase Cdk5, whose activity is largely restricted to the radial migration zone. Thus, the specific expression and Cdk5-dependent phosphorylation of RapGEF2 during multipolar-bipolar transition within the intermediate zone are essential for proper neuronal migration and wiring of the cerebral cortex.

  12. BSE Rap: intergenerational ties to save lives.

    PubMed

    Ehmann, J L

    1993-09-01

    This article presents an innovative public-education strategy that was created to promote breast health awareness and early breast cancer detection among minority and low-income adolescent females. Given the importance of teaching breast self-examination (BSE), program development focused on creation of the BSE Rap, a lively music-video presentation. Increasing adolescents' knowledge and awareness of BSE is viewed as a springboard for disseminating information to their mothers and grandmothers. Funding was obtained for production of a video and a breast health diary, which are the program's key components. Marketing strategies included contacts with community organizations and healthcare professionals. Program evaluations reveal that the BSE Rap serves as a positive motivator for participants to discuss BSE and mammography with their mothers and grandmothers. The BSE Rap offers oncology nurses the opportunity to save lives using a unique and creative tool that focuses on intergenerational ties. PMID:8415152

  13. Structure, functional regulation and signaling properties of Rap2B

    PubMed Central

    QU, DEBAO; HUANG, HUI; DI, JIEHUI; GAO, KEYU; LU, ZHENG; ZHENG, JUNNIAN

    2016-01-01

    The Ras family small guanosine 5′-triphosphate (GTP)-binding protein Rap2B is is a member of the Ras oncogene family and a novel target of p53 that regulates the p53-mediated pro-survival function of cells. The Rap2B protein shares ~90% homology with Rap2A, and its sequence is 70% identical to other members of the Rap family such as RaplA and RaplB. As a result, Rap2B has been theorized to have similar signaling effectors to the GTPase-binding protein Rap, which mediates various biological functions, including the regulation of sterile 20/mitogen-activated proteins. Since its identification in the early 1990s, Rap2B has elicited a considerable interest. Numerous studies indicate that Rap2B exerts specific biological functions, including binding and stimulating phospholipase C-ε and interferon-γ. In addition, downregulation of Rap2B affects the growth of melanoma cells. The present review summarizes the possible effectors and biological functions of Rap2B. Increasing evidence clearly supports the association between Rap2B function and tumor development. Therefore, it is conceivable that anticancer drugs targeting Rap2B may be generated as novel therapies against cancer. PMID:27073477

  14. Dialogic Teaching in an Online Environment: Book Raps

    ERIC Educational Resources Information Center

    Simpson, Alyson

    2010-01-01

    This paper examines a blended learning context known as book raps where children read and respond to literary texts. In this particular e-literacy environment, readers discuss their opinions of a book under the guidance of a moderator known as a rap coordinator who provides stimulus questions known as rap points. The paper demonstrates how…

  15. 40 CFR 270.68 - Remedial Action Plans (RAPs).

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false Remedial Action Plans (RAPs). 270.68 Section 270.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES... § 270.68 Remedial Action Plans (RAPs). Remedial Action Plans (RAPs) are special forms of permits...

  16. 40 CFR 270.68 - Remedial Action Plans (RAPs).

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false Remedial Action Plans (RAPs). 270.68 Section 270.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES... § 270.68 Remedial Action Plans (RAPs). Remedial Action Plans (RAPs) are special forms of permits...

  17. Cultural Studies and Rap: The Poetry of an Urban Lyricist

    ERIC Educational Resources Information Center

    Parmar, Priya

    2005-01-01

    In this article, the author explores the many other faces of rap that do not get the media exposure that they rightfully deserve. As this article attempts to reveal, rap music, as a form of cultural pedagogy and critical literacy, is only one way to achieve the goals of a "critical education." Rap lyrics can also be used as a tool to help the…

  18. 40 CFR 270.68 - Remedial Action Plans (RAPs).

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false Remedial Action Plans (RAPs). 270.68 Section 270.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES... § 270.68 Remedial Action Plans (RAPs). Remedial Action Plans (RAPs) are special forms of permits...

  19. 40 CFR 270.68 - Remedial Action Plans (RAPs).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Remedial Action Plans (RAPs). 270.68 Section 270.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES... § 270.68 Remedial Action Plans (RAPs). Remedial Action Plans (RAPs) are special forms of permits...

  20. 40 CFR 270.68 - Remedial Action Plans (RAPs).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false Remedial Action Plans (RAPs). 270.68 Section 270.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES... § 270.68 Remedial Action Plans (RAPs). Remedial Action Plans (RAPs) are special forms of permits...

  1. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false What is a RAP? 270.80 Section 270.80 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA... Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  2. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false What is a RAP? 270.80 Section 270.80 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA... Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  3. Modification of Caffeic Acid with Pyrrolidine Enhances Antioxidant Ability by Activating AKT/HO-1 Pathway in Heart

    PubMed Central

    Ku, Hui-Chun; Lee, Shih-Yi; Yang, Kai-Chien; Kuo, Yueh-Hsiung; Su, Ming-Jai

    2016-01-01

    Overproduction of free radicals during ischemia/reperfusion (I/R) injury leads to an interest in using antioxidant therapy. Activating an endogenous antioxidant signaling pathway is more important due to the fact that the free radical scavenging behavior in vitro does not always correlate with a cytoprotection effect in vivo. Caffeic acid (CA), an antioxidant, is a major phenolic constituent in nature. Pyrrolidinyl caffeamide (PLCA), a derivative of CA, was compared with CA for their antioxidant and cytoprotective effects. Our results indicate that CA and PLCA exert the same ability to scavenge DPPH in vitro. In response to myocardial I/R stress, PLCA was shown to attenuate lipid peroxydation and troponin release more than CA. These responses were accompanied with a prominent elevation in AKT and HO-1 expression and a preservation of mnSOD expression and catalase activity. PLCA also improved cell viability and alleviated the intracellular ROS level more than CA in cardiomyocytes exposed to H2O2. When inhibiting the AKT or HO-1 pathways, PLCA lost its ability to recover mnSOD expression and catalase activity to counteract with oxidative stress, suggesting AKT/HO-1 pathway activation by PLCA plays an important role. In addition, inhibition of AKT signaling further abolished HO-1 activity, while inhibition of HO-1 signaling attenuated AKT expression, indicating cross-talk between the AKT and HO-1 pathways. These protective effects may contribute to the cardiac function improvement by PLCA. These findings provide new insight into therapeutic approaches using a modified natural compound against oxidative stress from myocardial injuries. PMID:26845693

  4. Modification of Caffeic Acid with Pyrrolidine Enhances Antioxidant Ability by Activating AKT/HO-1 Pathway in Heart.

    PubMed

    Ku, Hui-Chun; Lee, Shih-Yi; Yang, Kai-Chien; Kuo, Yueh-Hsiung; Su, Ming-Jai

    2016-01-01

    Overproduction of free radicals during ischemia/reperfusion (I/R) injury leads to an interest in using antioxidant therapy. Activating an endogenous antioxidant signaling pathway is more important due to the fact that the free radical scavenging behavior in vitro does not always correlate with a cytoprotection effect in vivo. Caffeic acid (CA), an antioxidant, is a major phenolic constituent in nature. Pyrrolidinyl caffeamide (PLCA), a derivative of CA, was compared with CA for their antioxidant and cytoprotective effects. Our results indicate that CA and PLCA exert the same ability to scavenge DPPH in vitro. In response to myocardial I/R stress, PLCA was shown to attenuate lipid peroxydation and troponin release more than CA. These responses were accompanied with a prominent elevation in AKT and HO-1 expression and a preservation of mnSOD expression and catalase activity. PLCA also improved cell viability and alleviated the intracellular ROS level more than CA in cardiomyocytes exposed to H2O2. When inhibiting the AKT or HO-1 pathways, PLCA lost its ability to recover mnSOD expression and catalase activity to counteract with oxidative stress, suggesting AKT/HO-1 pathway activation by PLCA plays an important role. In addition, inhibition of AKT signaling further abolished HO-1 activity, while inhibition of HO-1 signaling attenuated AKT expression, indicating cross-talk between the AKT and HO-1 pathways. These protective effects may contribute to the cardiac function improvement by PLCA. These findings provide new insight into therapeutic approaches using a modified natural compound against oxidative stress from myocardial injuries. PMID:26845693

  5. TLR Signalling Pathways Diverge in Their Ability to Induce PGE2.

    PubMed

    Salvi, Valentina; Vaira, Xenia; Gianello, Veronica; Vermi, William; Bugatti, Mattia; Sozzani, Silvano; Bosisio, Daniela

    2016-01-01

    PGE2 is a lipid mediator abundantly produced in inflamed tissues that exerts relevant immunoregulatory functions. Dendritic cells (DCs) are key players in the onset and shaping of the inflammatory and immune responses and, as such, are well known PGE2 targets. By contrast, the precise role of human DCs in the production of PGE2 is poorly characterized. Here, we asked whether different ligands of Toll-like receptors (TLRs), a relevant family of pathogen-sensing receptors, could induce PGE2 in human DCs. The only active ligands were LPS (TLR4 ligand) and R848 (TLR7-8 ligand) although all TLRs, but TLR9, were expressed and functional. While investigating the molecular mechanisms hindering the release of PGE2, our experiments highlighted so far oversight differences in TLR signalling pathways in terms of MAPK and NF-κB activation. In addition, we identified that the PGE2-limiting checkpoint downstream TLR3, TLR5, and TLR7 was a defect in COX2 induction, while TLR1/2 and TLR2/6 failed to mobilize arachidonic acid, the substrate for the COX2 enzyme. Finally, we demonstrated the in vivo expression of PGE2 by myeloid CD11c(+) cells, documenting a role for DCs in the production of PGE2 in human inflamed tissues. PMID:27630451

  6. TLR Signalling Pathways Diverge in Their Ability to Induce PGE2

    PubMed Central

    Vaira, Xenia; Gianello, Veronica; Vermi, William; Bugatti, Mattia; Sozzani, Silvano

    2016-01-01

    PGE2 is a lipid mediator abundantly produced in inflamed tissues that exerts relevant immunoregulatory functions. Dendritic cells (DCs) are key players in the onset and shaping of the inflammatory and immune responses and, as such, are well known PGE2 targets. By contrast, the precise role of human DCs in the production of PGE2 is poorly characterized. Here, we asked whether different ligands of Toll-like receptors (TLRs), a relevant family of pathogen-sensing receptors, could induce PGE2 in human DCs. The only active ligands were LPS (TLR4 ligand) and R848 (TLR7-8 ligand) although all TLRs, but TLR9, were expressed and functional. While investigating the molecular mechanisms hindering the release of PGE2, our experiments highlighted so far oversight differences in TLR signalling pathways in terms of MAPK and NF-κB activation. In addition, we identified that the PGE2-limiting checkpoint downstream TLR3, TLR5, and TLR7 was a defect in COX2 induction, while TLR1/2 and TLR2/6 failed to mobilize arachidonic acid, the substrate for the COX2 enzyme. Finally, we demonstrated the in vivo expression of PGE2 by myeloid CD11c+ cells, documenting a role for DCs in the production of PGE2 in human inflamed tissues. PMID:27630451

  7. CDK5RAP2 is required for spindle checkpoint function.

    PubMed

    Zhang, Xiaoying; Liu, Dongyun; Lv, Shuang; Wang, Haibo; Zhong, Xueyan; Liu, Bo; Wang, Bo; Liao, Ji; Li, Jing; Pfeifer, Gerd P; Xu, Xingzhi

    2009-04-15

    The combination of paclitaxel and doxorubicin is among the most successful chemotherapy regimens in cancer treatment. CDK5RAP2, when mutated, causes primary microcephaly. We show here that inhibition of CDK5RAP2 expression causes chromosome mis-segregation, fails to maintain the spindle checkpoint, and is associated with reduced expression of the spindle checkpoint proteins BUBR1 and MAD2 and an increase in chromatin-associated CDC20. CDK5RAP2 resides on the BUBR1 and MAD2 promoters and regulates their transcription. Furthermore, CDK5RAP2-knockdown cells have increased resistance to paclitaxel and doxorubicin, and this resistance is partially rescued upon restoration of CDK5RAP2 expression. Cancer cells cultured in the presence of paclitaxel or doxorubicin exhibit dramatically decreased CDK5RAP2 levels. These results suggest that CDK5RAP2 is required for spindle checkpoint function and is a common target in paclitaxel and doxorubicin resistance. PMID:19282672

  8. System 80+ D-RAP, a communication tool

    SciTech Connect

    Siegmann, E.R.; Mody, A.A.

    1994-12-31

    The purpose of {open_quotes}RAP{close_quotes} music is to communicate, and the purpose of D-RAP is to foster communication between the probabilistic risk assessment (PRA) group, designers, and the future combined operating license (COL) applicant. This is to ensure that the design is self-consistent and integrated with the procurement process. The designer reliability assurance program (D-RAP) is the first part of the RAP. The goals of the D-RAP are to have risk-significant systems, structures, and components (SSCs) identified and considered in the detail design and procurement phases and to maintain consistency between PRA and design. Plant safety is maintained throughout the design phase, and pertinent information is passed on to the COL applicant. The operations RAP (O-RAP) covers the plant operation and maintenance.

  9. Rapping the 27 Amendments to the Constitution

    ERIC Educational Resources Information Center

    Knaresborough, Adam

    2009-01-01

    Early in the year, the students of history and government at Mountain View High School in Stafford, Virginia, began to devise hand motions to help memorize the 27 amendments to the Constitution for government class. Three students in the school who are interested in hip hop music then suggested composing a rap song about the topic. Working with…

  10. The Rap on Hip-Hop

    ERIC Educational Resources Information Center

    Piekarski, Bill

    2004-01-01

    From its humble origins some 30 years ago in New York's bombed-out, poverty-ravaged South Bronx, hip-hop has risen to become a dominant cultural force both here and abroad. Strictly defined, the term refers to the entire cultural constellation that accompanies rap music, which in 2001 surpassed country music as the most popular musical genre in…

  11. Structural Basis of Response Regulator Dephosphorylation by Rap Phosphatases

    SciTech Connect

    V Parashar; N Mirouze; D Dubnau; M Neiditch

    2011-12-31

    Bacterial Rap family proteins have been most extensively studied in Bacillus subtilis, where they regulate activities including sporulation, genetic competence, antibiotic expression, and the movement of the ICEBs1 transposon. One subset of Rap proteins consists of phosphatases that control B. subtilis and B. anthracis sporulation by dephosphorylating the response regulator Spo0F. The mechanistic basis of Rap phosphatase activity was unknown. Here we present the RapH-Spo0F X-ray crystal structure, which shows that Rap proteins consist of a 3-helix bundle and a tetratricopeptide repeat domain. Extensive biochemical and genetic functional studies reveal the importance of the observed RapH-Spo0F interactions, including the catalytic role of a glutamine in the RapH 3-helix bundle that inserts into the Spo0F active site. We show that in addition to dephosphorylating Spo0F, RapH can antagonize sporulation by sterically blocking phosphoryl transfer to and from Spo0F. Our structure-function analysis of the RapH-Spo0F interaction identified Rap protein residues critical for Spo0F phosphatase activity. This information enabled us to assign Spo0F phosphatase activity to a Rap protein based on sequence alone, which was not previously possible. Finally, as the ultimate test of our newfound understanding of the structural requirements for Rap phosphatase function, a non-phosphatase Rap protein that inhibits the binding of the response regulator ComA to DNA was rationally engineered to dephosphorylate Spo0F. In addition to revealing the mechanistic basis of response regulator dephosphorylation by Rap proteins, our studies support the previously proposed T-loop-Y allostery model of receiver domain regulation that restricts the aromatic 'switch' residue to an internal position when the {beta}4-{alpha}4 loop adopts an active-site proximal conformation.

  12. A Gα-Stimulated RapGEF Is a Receptor-Proximal Regulator of Dictyostelium Chemotaxis.

    PubMed

    Liu, Youtao; Lacal, Jesus; Veltman, Douwe M; Fusetti, Fabrizia; van Haastert, Peter J M; Firtel, Richard A; Kortholt, Arjan

    2016-06-01

    Chemotaxis, or directional movement toward extracellular chemical gradients, is an important property of cells that is mediated through G-protein-coupled receptors (GPCRs). Although many chemotaxis pathways downstream of Gβγ have been identified, few Gα effectors are known. Gα effectors are of particular importance because they allow the cell to distinguish signals downstream of distinct chemoattractant GPCRs. Here we identify GflB, a Gα2 binding partner that directly couples the Dictyostelium cyclic AMP GPCR to Rap1. GflB localizes to the leading edge and functions as a Gα-stimulated, Rap1-specific guanine nucleotide exchange factor required to balance Ras and Rap signaling. The kinetics of GflB translocation are fine-tuned by GSK-3 phosphorylation. Cells lacking GflB display impaired Rap1/Ras signaling and actin and myosin dynamics, resulting in defective chemotaxis. Our observations demonstrate that GflB is an essential upstream regulator of chemoattractant-mediated cell polarity and cytoskeletal reorganization functioning to directly link Gα activation to monomeric G-protein signaling. PMID:27237792

  13. RAP2.4a Is Transported through the Phloem to Regulate Cold and Heat Tolerance in Papaya Tree (Carica papaya cv. Maradol): Implications for Protection Against Abiotic Stress

    PubMed Central

    Arroyo-Herrera, Ana; Rodriguez-Corona, Ulises; Sanchez-Teyer, Felipe; Espadas-Alcocer, Jorge; Espadas-Gil, Francisco; Barredo-Pool, Felipe; Castaño, Enrique; Rodriguez-Zapata, Luis Carlos

    2016-01-01

    Plants respond to stress through metabolic and morphological changes that increase their ability to survive and grow. To this end, several transcription factor families are responsible for transmitting the signals that are required for these changes. Here, we studied the transcription factor superfamily AP2/ERF, particularly, RAP2.4 from Carica papaya cv. Maradol. We isolated four genes (CpRap2.4a, CpRAap2.4b, CpRap2.1 and CpRap2.10), and an in silico analysis showed that the four genes encode proteins that contain a conserved APETALA2 (AP2) domain located within group I and II transcription factors of the AP2/ERF superfamily. Semiquantitative PCR experiments indicated that each CpRap2 gene is differentially expressed under stress conditions, such as extreme temperatures. Moreover, genetic transformants of tobacco plants overexpressing CpRap2.4a and CpRap2.4b genes show a high level of tolerance to cold and heat stress compared to non-transformed plants. Confocal microscopy analysis of tobacco transgenic plants showed that CpRAP2.4a and CpRAP2.4b proteins were mainly localized to the nuclei of cells from the leaves and roots and also in the sieve elements. Moreover, the movement of CpRap2.4a RNA in tobacco grafting was analyzed. Our results indicate that CpRap2.4a and CpRap2.4b RNA in the papaya tree have a functional role in the response to stress conditions such as exposure to extreme temperatures via direct translation outside the parental RNA cell. PMID:27764197

  14. 40 CFR 270.95 - How do I apply for a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false How do I apply for a RAP? 270.95... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.95 How do I apply for a RAP? To apply for a RAP, you must complete an...

  15. 40 CFR 270.160 - When does my RAP become effective?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false When does my RAP become effective? 270... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.160 When does my RAP become effective? Your RAP becomes effective 30...

  16. 40 CFR 270.200 - How may I renew my RAP if it is expiring?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false How may I renew my RAP if it is... Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.200 How may I renew my RAP if it is expiring? If you wish to renew your expiring RAP, you must follow the process...

  17. 40 CFR 270.120 - To whom must I submit my RAP application?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false To whom must I submit my RAP... Action Plans (RAPs) Applying for A Rap § 270.120 To whom must I submit my RAP application? You must submit your application for a RAP to the Director for approval....

  18. 40 CFR 270.135 - What must the Director include in a draft RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... draft RAP? 270.135 Section 270.135 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Action Plans (RAPs) Getting A Rap Approved § 270.135 What must the Director include in a draft RAP? If the Director prepares a draft RAP, it must include the: (a) Information required under §...

  19. 40 CFR 270.200 - How may I renew my RAP if it is expiring?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false How may I renew my RAP if it is... Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.200 How may I renew my RAP if it is expiring? If you wish to renew your expiring RAP, you must follow the process...

  20. 40 CFR 270.210 - What records must I maintain concerning my RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... concerning my RAP? 270.210 Section 270.210 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.210 What records must I maintain concerning my RAP? You are required to keep records of: (a) All data used to complete RAP applications and...

  1. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... for a RAP? 270.110 Section 270.110 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Applying for A Rap § 270.110 What must I include in my application for a RAP? You must include the following information in your application for a RAP: (a) The name, address,...

  2. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... for a RAP? 270.110 Section 270.110 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Applying for A Rap § 270.110 What must I include in my application for a RAP? You must include the following information in your application for a RAP: (a) The name, address,...

  3. 40 CFR 270.210 - What records must I maintain concerning my RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... concerning my RAP? 270.210 Section 270.210 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.210 What records must I maintain concerning my RAP? You are required to keep records of: (a) All data used to complete RAP applications and...

  4. 40 CFR 270.95 - How do I apply for a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false How do I apply for a RAP? 270.95... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.95 How do I apply for a RAP? To apply for a RAP, you must complete an...

  5. 40 CFR 270.120 - To whom must I submit my RAP application?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false To whom must I submit my RAP... Action Plans (RAPs) Applying for A Rap § 270.120 To whom must I submit my RAP application? You must submit your application for a RAP to the Director for approval....

  6. 40 CFR 270.120 - To whom must I submit my RAP application?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false To whom must I submit my RAP... Action Plans (RAPs) Applying for A Rap § 270.120 To whom must I submit my RAP application? You must submit your application for a RAP to the Director for approval....

  7. 40 CFR 270.135 - What must the Director include in a draft RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... draft RAP? 270.135 Section 270.135 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Action Plans (RAPs) Getting A Rap Approved § 270.135 What must the Director include in a draft RAP? If the Director prepares a draft RAP, it must include the: (a) Information required under §...

  8. 40 CFR 270.200 - How may I renew my RAP if it is expiring?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false How may I renew my RAP if it is... Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.200 How may I renew my RAP if it is expiring? If you wish to renew your expiring RAP, you must follow the process...

  9. 40 CFR 270.160 - When does my RAP become effective?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false When does my RAP become effective? 270... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.160 When does my RAP become effective? Your RAP becomes effective 30...

  10. 40 CFR 270.160 - When does my RAP become effective?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false When does my RAP become effective? 270... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.160 When does my RAP become effective? Your RAP becomes effective 30...

  11. 40 CFR 270.135 - What must the Director include in a draft RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... draft RAP? 270.135 Section 270.135 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Action Plans (RAPs) Getting A Rap Approved § 270.135 What must the Director include in a draft RAP? If the Director prepares a draft RAP, it must include the: (a) Information required under §...

  12. 40 CFR 270.95 - How do I apply for a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false How do I apply for a RAP? 270.95... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.95 How do I apply for a RAP? To apply for a RAP, you must complete an...

  13. 40 CFR 270.120 - To whom must I submit my RAP application?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false To whom must I submit my RAP... Action Plans (RAPs) Applying for A Rap § 270.120 To whom must I submit my RAP application? You must submit your application for a RAP to the Director for approval....

  14. 40 CFR 270.120 - To whom must I submit my RAP application?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false To whom must I submit my RAP... Action Plans (RAPs) Applying for A Rap § 270.120 To whom must I submit my RAP application? You must submit your application for a RAP to the Director for approval....

  15. 40 CFR 270.210 - What records must I maintain concerning my RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... concerning my RAP? 270.210 Section 270.210 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.210 What records must I maintain concerning my RAP? You are required to keep records of: (a) All data used to complete RAP applications and...

  16. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... for a RAP? 270.110 Section 270.110 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Applying for A Rap § 270.110 What must I include in my application for a RAP? You must include the following information in your application for a RAP: (a) The name, address,...

  17. 40 CFR 270.210 - What records must I maintain concerning my RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... concerning my RAP? 270.210 Section 270.210 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.210 What records must I maintain concerning my RAP? You are required to keep records of: (a) All data used to complete RAP applications and...

  18. 40 CFR 270.135 - What must the Director include in a draft RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... draft RAP? 270.135 Section 270.135 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Action Plans (RAPs) Getting A Rap Approved § 270.135 What must the Director include in a draft RAP? If the Director prepares a draft RAP, it must include the: (a) Information required under §...

  19. 40 CFR 270.160 - When does my RAP become effective?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false When does my RAP become effective? 270... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.160 When does my RAP become effective? Your RAP becomes effective 30...

  20. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... for a RAP? 270.110 Section 270.110 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Applying for A Rap § 270.110 What must I include in my application for a RAP? You must include the following information in your application for a RAP: (a) The name, address,...

  1. 40 CFR 270.210 - What records must I maintain concerning my RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... concerning my RAP? 270.210 Section 270.210 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.210 What records must I maintain concerning my RAP? You are required to keep records of: (a) All data used to complete RAP applications and...

  2. 40 CFR 270.95 - How do I apply for a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false How do I apply for a RAP? 270.95... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.95 How do I apply for a RAP? To apply for a RAP, you must complete an...

  3. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... for a RAP? 270.110 Section 270.110 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Remedial Action Plans (RAPs) Applying for A Rap § 270.110 What must I include in my application for a RAP? You must include the following information in your application for a RAP: (a) The name, address,...

  4. 40 CFR 270.135 - What must the Director include in a draft RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... draft RAP? 270.135 Section 270.135 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Action Plans (RAPs) Getting A Rap Approved § 270.135 What must the Director include in a draft RAP? If the Director prepares a draft RAP, it must include the: (a) Information required under §...

  5. 40 CFR 270.95 - How do I apply for a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false How do I apply for a RAP? 270.95... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.95 How do I apply for a RAP? To apply for a RAP, you must complete an...

  6. 40 CFR 270.160 - When does my RAP become effective?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false When does my RAP become effective? 270... (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.160 When does my RAP become effective? Your RAP becomes effective 30...

  7. 40 CFR 270.200 - How may I renew my RAP if it is expiring?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false How may I renew my RAP if it is... Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.200 How may I renew my RAP if it is expiring? If you wish to renew your expiring RAP, you must follow the process...

  8. The Function of the Glutamate-Nitric Oxide-cGMP Pathway in Brain in Vivo and Learning Ability Decrease in Parallel in Mature Compared with Young Rats

    ERIC Educational Resources Information Center

    Piedrafita, Blanca; Cauli, Omar; Montoliu, Carmina; Felipo, Vicente

    2007-01-01

    Aging is associated with cognitive impairment, but the underlying mechanisms remain unclear. We have recently reported that the ability of rats to learn a Y-maze conditional discrimination task depends on the function of the glutamate-nitric oxide-cGMP pathway in brain. The aims of the present work were to assess whether the ability of rats to…

  9. Apical accumulation of the Sevenless receptor tyrosine kinase during Drosophila eye development is promoted by the small GTPase Rap1.

    PubMed

    Baril, Caroline; Lefrançois, Martin; Sahmi, Malha; Knævelsrud, Helene; Therrien, Marc

    2014-08-01

    The Ras/MAPK-signaling pathway plays pivotal roles during development of metazoans by controlling cell proliferation and cell differentiation elicited, in several instances, by receptor tyrosine kinases (RTKs). While the internal mechanism of RTK-driven Ras/MAPK signaling is well understood, far less is known regarding its interplay with other co-required signaling events involved in developmental decisions. In a genetic screen designed to identify new regulators of RTK/Ras/MAPK signaling during Drosophila eye development, we identified the small GTPase Rap1, PDZ-GEF, and Canoe as components contributing to Ras/MAPK-mediated R7 cell differentiation. Rap1 signaling has recently been found to participate in assembling cadherin-based adherens junctions in various fly epithelial tissues. Here, we show that Rap1 activity is required for the integrity of the apical domains of developing photoreceptor cells and that reduced Rap1 signaling hampers the apical accumulation of the Sevenless RTK in presumptive R7 cells. It thus appears that, in addition to its role in cell-cell adhesion, Rap1 signaling controls the partitioning of the epithelial cell membrane, which in turn influences signaling events that rely on apico-basal cell polarity.

  10. EPAC activation inhibits acetaldehyde-induced activation and proliferation of hepatic stellate cell via Rap1.

    PubMed

    Yang, Yan; Yang, Feng; Wu, Xiaojuan; Lv, Xiongwen; Li, Jun

    2016-05-01

    Hepatic stellate cells (HSCs) activation represents an essential event during alcoholic liver fibrosis (ALF). Previous studies have demonstrated that the rat HSCs could be significantly activated after exposure to 200 μmol/L acetaldehyde for 48 h, and the cAMP/PKA signaling pathways were also dramatically upregulated in activated HSCs isolated from alcoholic fibrotic rat liver. Exchange protein activated by cAMP (EPAC) is a family of guanine nucleotide exchange factors (GEFs) for the small Ras-like GTPases Rap, and is being considered as a vital mediator of cAMP signaling in parallel with the principal cAMP target protein kinase A (PKA). Our data showed that both cAMP/PKA and cAMP/EPAC signaling pathways were involved in acetaldehyde-induced HSCs. Acetaldehyde could reduce the expression of EPAC1 while enhancing the expression of EPAC2. The cAMP analog Me-cAMP, which stimulates the EPAC/Rap1 pathway, could significantly decrease the proliferation and collagen synthesis of acetaldehyde-induced HSCs. Furthermore, depletion of EPAC2, but not EPAC1, prevented the activation of HSC measured as the production of α-SMA and collagen type I and III, indicating that EPAC1 appears to have protective effects on acetaldehyde-induced HSCs. Curiously, activation of PKA or EPAC perhaps has opposite effects on the synthesis of collagen and α-SMA: EPAC activation by Me-cAMP increased the levels of GTP-bound (activated) Rap1 while PKA activation by Phe-cAMP had no significant effects on such binding. These results suggested that EPAC activation could inhibit the activation and proliferation of acetaldehyde-induced HSCs via Rap1. PMID:26854595

  11. Human Rap1 modulates TRF2 attraction to telomeric DNA

    PubMed Central

    Janoušková, Eliška; Nečasová, Ivona; Pavloušková, Jana; Zimmermann, Michal; Hluchý, Milan; Marini, Victoria; Nováková, Monika; Hofr, Ctirad

    2015-01-01

    More than two decades of genetic research have identified and assigned main biological functions of shelterin proteins that safeguard telomeres. However, a molecular mechanism of how each protein subunit contributes to the protecting function of the whole shelterin complex remains elusive. Human Repressor activator protein 1 (Rap1) forms a multifunctional complex with Telomeric Repeat binding Factor 2 (TRF2). Rap1–TRF2 complex is a critical part of shelterin as it suppresses homology-directed repair in Ku 70/80 heterodimer absence. To understand how Rap1 affects key functions of TRF2, we investigated full-length Rap1 binding to TRF2 and Rap1–TRF2 complex interactions with double-stranded DNA by quantitative biochemical approaches. We observed that Rap1 reduces the overall DNA duplex binding affinity of TRF2 but increases the selectivity of TRF2 to telomeric DNA. Additionally, we observed that Rap1 induces a partial release of TRF2 from DNA duplex. The improved TRF2 selectivity to telomeric DNA is caused by less pronounced electrostatic attractions between TRF2 and DNA in Rap1 presence. Thus, Rap1 prompts more accurate and selective TRF2 recognition of telomeric DNA and TRF2 localization on single/double-strand DNA junctions. These quantitative functional studies contribute to the understanding of the selective recognition of telomeric DNA by the whole shelterin complex. PMID:25675958

  12. Rap Music Literacy: A Case Study of Millennial Audience Reception to Rap Lyrics Depicting Independent Women

    ERIC Educational Resources Information Center

    Moody-Ramirez, Mia; Scott, Lakia M.

    2015-01-01

    Using a feminist lens and a constructivist approach as the theoretical framework, we used rap lyrics and videos to help college students explore mass media's representation of the "independent" Black woman and the concept of "independence" in general. Students must be able to formulate their own concept of independence to…

  13. Oxygen Sensing via the Ethylene Response Transcription Factor RAP2.12 Affects Plant Metabolism and Performance under Both Normoxia and Hypoxia.

    PubMed

    Paul, Melanie Verena; Iyer, Srignanakshi; Amerhauser, Carmen; Lehmann, Martin; van Dongen, Joost T; Geigenberger, Peter

    2016-09-01

    Subgroup-VII-ethylene-response-factor (ERF-VII) transcription factors are involved in the regulation of hypoxic gene expression and regulated by proteasome-mediated proteolysis via the oxygen-dependent branch of the N-end-rule pathway. While research into ERF-VII mainly focused on their role to regulate anoxic gene expression, little is known on the impact of this oxygen-sensing system in regulating plant metabolism and growth. By comparing Arabidopsis (Arabidopsis thaliana) plants overexpressing N-end-rule-sensitive and insensitive forms of the ERF-VII-factor RAP2.12, we provide evidence that oxygen-dependent RAP2.12 stability regulates central metabolic processes to sustain growth, development, and anoxic resistance of plants. (1) Under normoxia, overexpression of N-end-rule-insensitive Δ13RAP2.12 led to increased activities of fermentative enzymes and increased accumulation of fermentation products, which were accompanied by decreased adenylate energy states and starch levels, and impaired plant growth and development, indicating a role of oxygen-regulated RAP2.12 degradation to prevent aerobic fermentation. (2) In Δ13RAP2.12-overexpressing plants, decreased carbohydrate reserves also led to a decrease in anoxic resistance, which was prevented by external Suc supply. (3) Overexpression of Δ13RAP2.12 led to decreased respiration rates, changes in the levels of tricarboxylic acid cycle intermediates, and accumulation of a large number of amino acids, including Ala and γ-amino butyric acid, indicating a role of oxygen-regulated RAP2.12 abundance in controlling the flux-modus of the tricarboxylic acid cycle. (4) The increase in amino acids was accompanied by increased levels of immune-regulatory metabolites. These results show that oxygen-sensing, mediating RAP2.12 degradation is indispensable to optimize metabolic performance, plant growth, and development under both normoxic and hypoxic conditions. PMID:27372243

  14. A plasmid-born Rap-Phr system regulates surfactin production, sporulation and genetic competence in the heterologous host, Bacillus subtilis OKB105.

    PubMed

    Yang, Yang; Wu, Hui-Jun; Lin, Ling; Zhu, Qing-Qing; Borriss, Rainer; Gao, Xue-Wen

    2015-09-01

    According to the change of environment, soil-dwelling Bacillus species differentiate into distinct subpopulations, such as spores and competent cells. Rap-Phr systems have been found to be involved in this differentiation circuit by interacting with major regulatory proteins, such as Spo0A, ComA, and DegU. In this study, we report that the plasmid-born RapQ-PhrQ system found in Bacillus amyloliquefaciens B3 affects three regulatory pathways in the heterologous host Bacillus subtilis. Expression of rapQ in B. subtilis OKB105 strongly suppressed its sporulation efficiency, transformation efficiency, and surfactin production. Co-expression of phrQ or addition of synthesized PhrQ pentapeptide in vitro could compensate for the suppressive effects caused by rapQ. We also found that expression of rapQ decreased the transcriptional level of the sporulation-related gene spoIIE and surfactin synthesis-related gene srfA; meanwhile, the transcriptional levels of these genes could be rescued by co-expression of phrQ and in vitro addition of PhrQ pentapeptide. Electrophoretic mobility shift (EMSA) result also showed that RapQ could bind to ComA without interacting with ComA binding to DNA, and PhrQ pentapeptide antagonized RapQ activity in vitro. These results indicate that this new plasmid-born RapQ-PhrQ system controls sporulation, competent cell formation, and surfactin production in B. subtilis OKB105.

  15. Oxygen Sensing via the Ethylene Response Transcription Factor RAP2.12 Affects Plant Metabolism and Performance under Both Normoxia and Hypoxia1[OPEN

    PubMed Central

    Paul, Melanie Verena; Iyer, Srignanakshi; Lehmann, Martin

    2016-01-01

    Subgroup-VII-ethylene-response-factor (ERF-VII) transcription factors are involved in the regulation of hypoxic gene expression and regulated by proteasome-mediated proteolysis via the oxygen-dependent branch of the N-end-rule pathway. While research into ERF-VII mainly focused on their role to regulate anoxic gene expression, little is known on the impact of this oxygen-sensing system in regulating plant metabolism and growth. By comparing Arabidopsis (Arabidopsis thaliana) plants overexpressing N-end-rule-sensitive and insensitive forms of the ERF-VII-factor RAP2.12, we provide evidence that oxygen-dependent RAP2.12 stability regulates central metabolic processes to sustain growth, development, and anoxic resistance of plants. (1) Under normoxia, overexpression of N-end-rule-insensitive Δ13RAP2.12 led to increased activities of fermentative enzymes and increased accumulation of fermentation products, which were accompanied by decreased adenylate energy states and starch levels, and impaired plant growth and development, indicating a role of oxygen-regulated RAP2.12 degradation to prevent aerobic fermentation. (2) In Δ13RAP2.12-overexpressing plants, decreased carbohydrate reserves also led to a decrease in anoxic resistance, which was prevented by external Suc supply. (3) Overexpression of Δ13RAP2.12 led to decreased respiration rates, changes in the levels of tricarboxylic acid cycle intermediates, and accumulation of a large number of amino acids, including Ala and γ-amino butyric acid, indicating a role of oxygen-regulated RAP2.12 abundance in controlling the flux-modus of the tricarboxylic acid cycle. (4) The increase in amino acids was accompanied by increased levels of immune-regulatory metabolites. These results show that oxygen-sensing, mediating RAP2.12 degradation is indispensable to optimize metabolic performance, plant growth, and development under both normoxic and hypoxic conditions. PMID:27372243

  16. Listening to Rap: Cultures of Crime, Cultures of Resistance

    ERIC Educational Resources Information Center

    Tanner, Julian; Asbridge, Mark; Wortley, Scot

    2009-01-01

    This research compares representations of rap music with the self-reported criminal behavior and resistant attitudes of the music's core audience. Our database is a large sample of Toronto high school students (n = 3,393) from which we identify a group of listeners, whose combination of musical likes and dislikes distinguish them as rap univores.…

  17. Simultaneous functions of the installed DAS/DAK formaldehyde-assimilation pathway and the original formaldehyde metabolic pathways enhance the ability of transgenic geranium to purify gaseous formaldehyde polluted environment.

    PubMed

    Zhou, Shengen; Xiao, Sunqin; Xuan, Xiuxia; Sun, Zhen; Li, Kunzhi; Chen, Limei

    2015-04-01

    The overexpression of dihydroxyacetone synthase (DAS) and dihydroxyacetone kinase (DAK) from methylotrophic yeasts in chloroplasts created a photosynthetic formaldehyde (HCHO)-assimilation pathway (DAS/DAK pathway) in transgenic tobacco. Geranium has abilities to absorb and metabolize HCHO. Results of this study showed that the installed DAS/DAK pathway functioning in chloroplasts greatly enhanced the role of the Calvin cycle in transgenic geranium under high concentrations of gaseous HCHO stress. Consequently, the yield of sugars from HCHO-assimilation increased approximately 6-fold in transgenic geranium leaves, and concomitantly, the role of three original HCHO metabolic pathways reduced, leading to a significant decrease in formic acid, citrate and glycine production from HCHO metabolism. Although the role of three metabolic pathways reduced in transgenic plants under high concentrations of gaseous HCHO stress, the installed DAS/DAK pathway could still function together with the original HCHO metabolic pathways. Consequently, the gaseous HCHO-resistance of transgenic plants was significantly improved, and the generation of H2O2 in the transgenic geranium leaves was significantly less than that in the wild type (WT) leaves. Under environmental-polluted gaseous HCHO stress for a long duration, the stomata conductance of transgenic plants remained approximately 2-fold higher than that of the WT, thereby increasing its ability to purify gaseous HCHO polluted environment.

  18. Simultaneous functions of the installed DAS/DAK formaldehyde-assimilation pathway and the original formaldehyde metabolic pathways enhance the ability of transgenic geranium to purify gaseous formaldehyde polluted environment.

    PubMed

    Zhou, Shengen; Xiao, Sunqin; Xuan, Xiuxia; Sun, Zhen; Li, Kunzhi; Chen, Limei

    2015-04-01

    The overexpression of dihydroxyacetone synthase (DAS) and dihydroxyacetone kinase (DAK) from methylotrophic yeasts in chloroplasts created a photosynthetic formaldehyde (HCHO)-assimilation pathway (DAS/DAK pathway) in transgenic tobacco. Geranium has abilities to absorb and metabolize HCHO. Results of this study showed that the installed DAS/DAK pathway functioning in chloroplasts greatly enhanced the role of the Calvin cycle in transgenic geranium under high concentrations of gaseous HCHO stress. Consequently, the yield of sugars from HCHO-assimilation increased approximately 6-fold in transgenic geranium leaves, and concomitantly, the role of three original HCHO metabolic pathways reduced, leading to a significant decrease in formic acid, citrate and glycine production from HCHO metabolism. Although the role of three metabolic pathways reduced in transgenic plants under high concentrations of gaseous HCHO stress, the installed DAS/DAK pathway could still function together with the original HCHO metabolic pathways. Consequently, the gaseous HCHO-resistance of transgenic plants was significantly improved, and the generation of H2O2 in the transgenic geranium leaves was significantly less than that in the wild type (WT) leaves. Under environmental-polluted gaseous HCHO stress for a long duration, the stomata conductance of transgenic plants remained approximately 2-fold higher than that of the WT, thereby increasing its ability to purify gaseous HCHO polluted environment. PMID:25698666

  19. Rap Music Genres and Deviant Behaviors in French-Canadian Adolescents

    ERIC Educational Resources Information Center

    Miranda, Dave; Claes, Michel

    2004-01-01

    This study investigated the links between the preference for 4 rap music genres (American rap, French rap, hip hop/soul, and gangsta/hardcore rap) and 5 types of deviant behaviors in adolescence (violence, theft, street gangs, mild drug use, and hard drug use). The effects of peers' deviancy, violent media, and importance given to lyrics were…

  20. Phospholipase Cε Modulates Rap1 Activity and the Endothelial Barrier

    PubMed Central

    DiStefano, Peter V.; Smrcka, Alan V.; Glading, Angela J.

    2016-01-01

    The phosphoinositide-specific phospholipase C, PLCε, is a unique signaling protein with known roles in regulating cardiac myocyte growth, astrocyte inflammatory signaling, and tumor formation. PLCε is also expressed in endothelial cells, however its role in endothelial regulation is not fully established. We show that endothelial cells of multiple origins, including human pulmonary artery (HPAEC), human umbilical vein (HUVEC), and immortalized brain microvascular (hCMEC/D3) endothelial cells, express PLCε. Knockdown of PLCε in arterial endothelial monolayers decreased the effectiveness of the endothelial barrier. Concomitantly, RhoA activity and stress fiber formation were increased. PLCε-deficient arterial endothelial cells also exhibited decreased Rap1-GTP levels, which could be restored by activation of the Rap1 GEF, Epac, to rescue the increase in monolayer leak. Reintroduction of PLCε rescued monolayer leak with both the CDC25 GEF domain and the lipase domain of PLCε required to fully activate Rap1 and to rescue endothelial barrier function. Finally, we demonstrate that the barrier promoting effects PLCε are dependent on Rap1 signaling through the Rap1 effector, KRIT1, which we have previously shown is vital for maintaining endothelial barrier stability. Thus we have described a novel role for PLCε PIP2 hydrolytic and Rap GEF activities in arterial endothelial cells, where PLCε-dependent activation of Rap1/KRIT1 signaling promotes endothelial barrier stability. PMID:27612188

  1. Phospholipase Cε Modulates Rap1 Activity and the Endothelial Barrier.

    PubMed

    DiStefano, Peter V; Smrcka, Alan V; Glading, Angela J

    2016-01-01

    The phosphoinositide-specific phospholipase C, PLCε, is a unique signaling protein with known roles in regulating cardiac myocyte growth, astrocyte inflammatory signaling, and tumor formation. PLCε is also expressed in endothelial cells, however its role in endothelial regulation is not fully established. We show that endothelial cells of multiple origins, including human pulmonary artery (HPAEC), human umbilical vein (HUVEC), and immortalized brain microvascular (hCMEC/D3) endothelial cells, express PLCε. Knockdown of PLCε in arterial endothelial monolayers decreased the effectiveness of the endothelial barrier. Concomitantly, RhoA activity and stress fiber formation were increased. PLCε-deficient arterial endothelial cells also exhibited decreased Rap1-GTP levels, which could be restored by activation of the Rap1 GEF, Epac, to rescue the increase in monolayer leak. Reintroduction of PLCε rescued monolayer leak with both the CDC25 GEF domain and the lipase domain of PLCε required to fully activate Rap1 and to rescue endothelial barrier function. Finally, we demonstrate that the barrier promoting effects PLCε are dependent on Rap1 signaling through the Rap1 effector, KRIT1, which we have previously shown is vital for maintaining endothelial barrier stability. Thus we have described a novel role for PLCε PIP2 hydrolytic and Rap GEF activities in arterial endothelial cells, where PLCε-dependent activation of Rap1/KRIT1 signaling promotes endothelial barrier stability. PMID:27612188

  2. Ensemble covariances adaptively localized with ECO-RAP. Part 2: a strategy for the atmosphere

    NASA Astrophysics Data System (ADS)

    Bishop, Craig H.; Hodyss, Daniel

    2009-01-01

    Part 1's localization method, Ensemble COrrelations Raised to A Power (ECO-RAP), is incorporated into a Local Ensemble Transform Kalman Filter (LETKF). Because brute force incorporation would be too expensive, we demonstrate a factorization property for Part 1's Covariances Adaptively Localized with ECO-rap (CALECO) forecast error covariance matrix that, together with other simplifications, reduces the cost. The property inexpensively provides a large CALECO ensemble whose covariance is the CALECO matrix. Each member of the CALECO ensemble is an element-wise product between one raw ensemble member and one column of the square root of the ECO-RAP matrix. The LETKF is applied to the CALECO ensemble rather than the raw ensemble. The approach enables the update of large numbers of variables within each observation volume at little additional computational cost. Under plausible assumptions, this makes the CALECO and standard LETKF costs similar. The CALECO LETKF does not require artificial observation error inflation or vertically confined observation volumes both of which confound the assimilation of non-local observations such as satellite observations. Using a 27 member ensemble from a global Numerical Weather Prediction (NWP) system, we depict four-dimensional (4-D) flow-adaptive error covariance localization and test the ability of the CALECO LETKF to reduce analysis error.

  3. 40 CFR 270.105 - Who must sign the application and any required reports for a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... required reports for a RAP? 270.105 Section 270.105 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.105 Who must sign the application and any required reports for a RAP? Both the owner and the operator must sign the RAP application and any...

  4. 40 CFR 270.125 - If I submit my RAP application as part of another document, what must I do?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false If I submit my RAP application as part... PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.125 If I submit my RAP application as part of another document, what must I do? If you submit your application for a RAP as a part...

  5. 40 CFR 270.105 - Who must sign the application and any required reports for a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... required reports for a RAP? 270.105 Section 270.105 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.105 Who must sign the application and any required reports for a RAP? Both the owner and the operator must sign the RAP application and any...

  6. 40 CFR 270.125 - If I submit my RAP application as part of another document, what must I do?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false If I submit my RAP application as part... PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.125 If I submit my RAP application as part of another document, what must I do? If you submit your application for a RAP as a part...

  7. 40 CFR 270.125 - If I submit my RAP application as part of another document, what must I do?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false If I submit my RAP application as part... PERMIT PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.125 If I submit my RAP application as part of another document, what must I do? If you submit your application for a RAP as a part...

  8. 40 CFR 270.105 - Who must sign the application and any required reports for a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... required reports for a RAP? 270.105 Section 270.105 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.105 Who must sign the application and any required reports for a RAP? Both the owner and the operator must sign the RAP application and any...

  9. 40 CFR 270.105 - Who must sign the application and any required reports for a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... required reports for a RAP? 270.105 Section 270.105 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.105 Who must sign the application and any required reports for a RAP? Both the owner and the operator must sign the RAP application and any...

  10. 40 CFR 270.105 - Who must sign the application and any required reports for a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... required reports for a RAP? 270.105 Section 270.105 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Applying for A Rap § 270.105 Who must sign the application and any required reports for a RAP? Both the owner and the operator must sign the RAP application and any...

  11. cAMP blocks MAPK activation and sclerotial development via Rap-1 in a PKA-independent manner in Sclerotinia sclerotiorum.

    PubMed

    Chen, Changbin; Dickman, Martin B

    2005-01-01

    Sclerotinia sclerotiorum is a filamentous ascomycete phytopathogen able to infect an extremely wide range of cultivated plants. Our previous studies have shown that increases in cAMP levels result in the impairment of the development of the sclerotium, a highly differentiated structure important in the disease cycle of this fungus. cAMP also inhibits the activation of a S. sclerotiorum mitogen-activated protein kinase (MAPK), which we have previously shown to be required for sclerotial maturation; thus cAMP-mediated sclerotial inhibition is modulated through MAPK. However, the mechanism(s) by which cAMP inhibits MAPK remains unclear. Here we demonstrate that a protein kinase A (PKA)-independent signalling pathway probably mediates MAPK inhibition by cAMP. Expression of a dominant negative form of Ras, an upstream activator of the MAPK pathway, also inhibited sclerotial development and MAPK activation, suggesting that a conserved Ras/MAPK pathway is required for sclerotial development. Evidence from bacterial toxins that specifically inhibit the activity of small GTPases, suggested that Rap-1 or Ras is involved in cAMP action. The Rap-1 inhibitor, GGTI-298, restored MAPK activation in the presence of cAMP, further suggesting that Rap-1 is responsible for cAMP-dependent MAPK inhibition. Importantly, inhibition of Rap-1 is able to restore sclerotial development blocked by cAMP. Our results suggest a novel mechanism involving the requirement of Ras/MAPK pathway for sclerotial development that is negatively regulated by a PKA-independent cAMP signalling pathway. Cross-talk between these two pathways is mediated by Rap-1. PMID:15612936

  12. Optimization of the rapping process of an intermittent electrostatic precipitator

    NASA Astrophysics Data System (ADS)

    Miloua, F.; Tilmatine, A.; Gouri, R.; Kadous, N.; Dascalescu, L.

    2008-01-01

    Intermittent operation mode is specific to electrostatic precipitators (ESP) used in workshops where the polluting product is produced in a discontinuous way. The rapping system is necessary in order to ensure a continuous and effective operation of a dry electrostatic precipitator, but causes at the same time, a problem of re-entrainment of dust and thus the degradation of filtration efficiency. The objective of this paper is to propose a procedure based on the methodology of experimental designs (Tagushi's Methodology) aiming at optimizing the rapping process; it consists to determine optimal values of rapping, i.e. the moment, the position and the force of rapping. Several “one-factor-at-a-time" experimental designs followed by a Full Factorial design, made it possible to model the process and to analyze interactions between the factors. The experiments were carried out on a laboratory experimental device which simulates an industrial precipitator with intermittent operation.

  13. Chaperone-mediated specificity in Ras and Rap signaling.

    PubMed

    Azoulay-Alfaguter, Inbar; Strazza, Marianne; Mor, Adam

    2015-01-01

    Ras and Rap proteins are closely related small guanosine triphosphatase (GTPases) that share similar effector-binding domains but operate in a very different signaling networks; Ras has a dominant role in cell proliferation, while Rap mediates cell adhesion. Ras and Rap proteins are regulated by several shared processes such as post-translational modification, phosphorylation, activation by guanine exchange factors and inhibition by GTPase-activating proteins. Sub-cellular localization and trafficking of these proteins to and from the plasma membrane are additional important regulatory features that impact small GTPases function. Despite its importance, the trafficking mechanisms of Ras and Rap proteins are not completely understood. Chaperone proteins play a critical role in trafficking of GTPases and will be the focus of the discussion in this work. We will review several aspects of chaperone biology focusing on specificity toward particular members of the small GTPase family. Understanding this specificity should provide key insights into drug development targeting individual small GTPases.

  14. RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia

    PubMed Central

    Pascoal-Xavier, Marcelo Antonio; Figueiredo, Anna Carolina Cançado; Gomes, Luciana Inácia; Peruhype-Magalhães, Vanessa; Calzavara-Silva, Carlos Eduardo; Costa, Marcelo Azevedo; Reis, Ilka Afonso; Bonjardim, Claudio Antônio; Kroon, Erna Geessien

    2015-01-01

    RAP1 (RAS proximate 1), a small GTP-binding protein of the RAS superfamily, is a putative oncogene that is highly expressed in several malignant cell lines and types of cancers, including some types of squamous cell carcinoma. However, the participation of RAP1 in cervical carcinogenesis is unknown. We conducted a cross-sectional study of paraffin-embedded cervical biopsies to determine the association of RAP1 with cervical intraepithelial neoplasia (CIN). Standard and quantitative immunohistochemistry assessment of RAP1 expression in fixed tissue was performed on 183 paraffin-embedded cervical biopsies that were classified as normal or non-dysplastic mucosa (NDM) (n = 33); CIN grade 1 (n = 84) and CIN grade 2/3 (n = 66). A gradual increase in RAP1 expression in NDM < CIN 1 < CIN 2/3 (p<0.001) specimens was observed and was in agreement with the histopathologic diagnosis. A progressive increase in the RAP1 expression levels increased the risk of CIN 1 [odds ratio (OR) = 3.50; 95% confidence interval (CI) 1.30-10.64] 3.5 fold and the risk of CIN 2/3 (OR = 19.86, 95% CI 6.40-70.79) nearly 20 fold when compared to NDM. In addition, stereotype ordinal regression analysis showed that this progressive increase in RAP1 expression more strongly impacted CIN 2/3 than CIN 1. Our findings suggest that RAP1 may be a useful biomarker for the diagnosis of CIN. PMID:25856570

  15. Mutations in CDK5RAP2 cause Seckel syndrome

    PubMed Central

    Yigit, Gökhan; Brown, Karen E; Kayserili, Hülya; Pohl, Esther; Caliebe, Almuth; Zahnleiter, Diana; Rosser, Elisabeth; Bögershausen, Nina; Uyguner, Zehra Oya; Altunoglu, Umut; Nürnberg, Gudrun; Nürnberg, Peter; Rauch, Anita; Li, Yun; Thiel, Christian Thomas; Wollnik, Bernd

    2015-01-01

    Seckel syndrome is a heterogeneous, autosomal recessive disorder marked by prenatal proportionate short stature, severe microcephaly, intellectual disability, and characteristic facial features. Here, we describe the novel homozygous splice-site mutations c.383+1G>C and c.4005-9A>G in CDK5RAP2 in two consanguineous families with Seckel syndrome. CDK5RAP2 (CEP215) encodes a centrosomal protein which is known to be essential for centrosomal cohesion and proper spindle formation and has been shown to be causally involved in autosomal recessive primary microcephaly. We establish CDK5RAP2 as a disease-causing gene for Seckel syndrome and show that loss of functional CDK5RAP2 leads to severe defects in mitosis and spindle organization, resulting in cells with abnormal nuclei and centrosomal pattern, which underlines the important role of centrosomal and mitotic proteins in the pathogenesis of the disease. Additionally, we present an intriguing case of possible digenic inheritance in Seckel syndrome: A severely affected child of nonconsanguineous German parents was found to carry heterozygous mutations in CDK5RAP2 and CEP152. This finding points toward a potential additive genetic effect of mutations in CDK5RAP2 and CEP152. PMID:26436113

  16. Mutations in CDK5RAP2 cause Seckel syndrome.

    PubMed

    Yigit, Gökhan; Brown, Karen E; Kayserili, Hülya; Pohl, Esther; Caliebe, Almuth; Zahnleiter, Diana; Rosser, Elisabeth; Bögershausen, Nina; Uyguner, Zehra Oya; Altunoglu, Umut; Nürnberg, Gudrun; Nürnberg, Peter; Rauch, Anita; Li, Yun; Thiel, Christian Thomas; Wollnik, Bernd

    2015-09-01

    Seckel syndrome is a heterogeneous, autosomal recessive disorder marked by prenatal proportionate short stature, severe microcephaly, intellectual disability, and characteristic facial features. Here, we describe the novel homozygous splice-site mutations c.383+1G>C and c.4005-9A>G in CDK5RAP2 in two consanguineous families with Seckel syndrome. CDK5RAP2 (CEP215) encodes a centrosomal protein which is known to be essential for centrosomal cohesion and proper spindle formation and has been shown to be causally involved in autosomal recessive primary microcephaly. We establish CDK5RAP2 as a disease-causing gene for Seckel syndrome and show that loss of functional CDK5RAP2 leads to severe defects in mitosis and spindle organization, resulting in cells with abnormal nuclei and centrosomal pattern, which underlines the important role of centrosomal and mitotic proteins in the pathogenesis of the disease. Additionally, we present an intriguing case of possible digenic inheritance in Seckel syndrome: A severely affected child of nonconsanguineous German parents was found to carry heterozygous mutations in CDK5RAP2 and CEP152. This finding points toward a potential additive genetic effect of mutations in CDK5RAP2 and CEP152.

  17. Mutations in CDK5RAP2 cause Seckel syndrome.

    PubMed

    Yigit, Gökhan; Brown, Karen E; Kayserili, Hülya; Pohl, Esther; Caliebe, Almuth; Zahnleiter, Diana; Rosser, Elisabeth; Bögershausen, Nina; Uyguner, Zehra Oya; Altunoglu, Umut; Nürnberg, Gudrun; Nürnberg, Peter; Rauch, Anita; Li, Yun; Thiel, Christian Thomas; Wollnik, Bernd

    2015-09-01

    Seckel syndrome is a heterogeneous, autosomal recessive disorder marked by prenatal proportionate short stature, severe microcephaly, intellectual disability, and characteristic facial features. Here, we describe the novel homozygous splice-site mutations c.383+1G>C and c.4005-9A>G in CDK5RAP2 in two consanguineous families with Seckel syndrome. CDK5RAP2 (CEP215) encodes a centrosomal protein which is known to be essential for centrosomal cohesion and proper spindle formation and has been shown to be causally involved in autosomal recessive primary microcephaly. We establish CDK5RAP2 as a disease-causing gene for Seckel syndrome and show that loss of functional CDK5RAP2 leads to severe defects in mitosis and spindle organization, resulting in cells with abnormal nuclei and centrosomal pattern, which underlines the important role of centrosomal and mitotic proteins in the pathogenesis of the disease. Additionally, we present an intriguing case of possible digenic inheritance in Seckel syndrome: A severely affected child of nonconsanguineous German parents was found to carry heterozygous mutations in CDK5RAP2 and CEP152. This finding points toward a potential additive genetic effect of mutations in CDK5RAP2 and CEP152. PMID:26436113

  18. RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli

    PubMed Central

    Zampini, Massimiliano; Stevens, Pauline Rees; Pachebat, Justin A.; Kingston-Smith, Alison; Mur, Luis A. J.; Hayes, Finbarr

    2015-01-01

    The ability to assemble DNA sequences de novo through efficient and powerful DNA fabrication methods is one of the foundational technologies of synthetic biology. Gene synthesis, in particular, has been considered the main driver for the emergence of this new scientific discipline. Here we describe RapGene, a rapid gene assembly technique which was successfully tested for the synthesis and cloning of both prokaryotic and eukaryotic genes through a ligation independent approach. The method developed in this study is a complete bacterial gene synthesis platform for the quick, accurate and cost effective fabrication and cloning of gene-length sequences that employ the widely used host Escherichia coli. PMID:26062748

  19. RAP-011 augments callus formation in closed fractures in rats.

    PubMed

    Morse, Alyson; Cheng, Tegan L; Peacock, Lauren; Mikulec, Kathy; Little, David G; Schindeler, Aaron

    2016-02-01

    ACE-011 is a bone anabolic agent generated by fusing the extracellular domain of the Activin Type 2A receptor (ActRIIA) to an IgG-Fc. The orthopedic utility of ACE-011 was investigated using a murine analogue, RAP-011. Initially, a rat closed fracture model was tested using bi-weekly (biw) 10 mg/kg RAP-011. RAP-011 significantly increased callus length and callus bone volume (BV, +43% at 6w, p < 0.01). The polar moment of inertia was calculated to be substantively increased (+80%, p < 0.01), however mechanical bending tests showed a more modest increase in maximum load to failure (+24%, p < 0.05). Histology indicated enhanced appositional bone growth, but it was hypothesized that reduced remodeling, evidenced by decreased serum CTX (-16% at 6w, p < 0.01), could be compromising bone quality in the callus. A second closed fracture study was performed to examine lower "pulse" [RAP-011(p)] and "sustained" [RAP-011(s)] regimens of biw 0.6mg/kg × 2, 0.35mg/kg × 3 and 0.18mg/kg × 2, 0.1mg/kg × 7 respectively, compared with PTH(1-34) (25 μg/kg/d) and vehicle controls. RAP-011 treatments gave modest increases in callus length and callus BV at 6w (p < 0.01), but did not achieve an increase in maximum load over vehicle. In summary, RAP-011 is effective in promoting bone formation during repair, but optimizing callus bone quality will require further investigation.

  20. Remote Area Power Supply (RAPS) load and resource profiles.

    SciTech Connect

    Giles, Lauren; Skolnik, Edward G.; Marchionini, Brian; Fall, Ndeye K.

    2007-07-01

    In 1997, an international team interested in the development of Remote Area Power Supply (RAPS) systems for rural electrification projects around the world was organized by the International Lead Zinc Research Organization (ILZRO) with the support of Sandia National Laboratories (SNL). The team focused on defining load and resource profiles for RAPS systems. They identified single family homes, small communities, and villages as candidates for RAPS applications, and defined several different size/power requirements for each. Based on renewable energy and resource data, the team devised a ''strawman'' series of load profiles. A RAPS system typically consists of a renewable and/or conventional generator, power conversion equipment, and a battery. The purpose of this report is to present data and information on insolation levels and load requirements for ''typical'' homes, small communities, and larger villages around the world in order to facilitate the development of robust design practices for RAPS systems, and especially for the storage battery component. These systems could have significant impact on areas of the world that would otherwise not be served by conventional electrical grids.

  1. Epac Activates the Small G Proteins Rap1 and Rab3A to Achieve Exocytosis*

    PubMed Central

    Branham, María T.; Bustos, Matías A.; De Blas, Gerardo A.; Rehmann, Holger; Zarelli, Valeria E. P.; Treviño, Claudia L.; Darszon, Alberto; Mayorga, Luis S.; Tomes, Claudia N.

    2009-01-01

    Exocytosis of the acrosome (the acrosome reaction) relies on cAMP production, assembly of a proteinaceous fusion machinery, calcium influx from the extracellular medium, and mobilization from inositol 1,4,5-trisphosphate-sensitive intracellular stores. Addition of cAMP to human sperm suspensions bypasses some of these requirements and elicits exocytosis in a protein kinase A- and extracellular calcium-independent manner. The relevant cAMP target is Epac, a guanine nucleotide exchange factor for the small GTPase Rap. We show here that a soluble adenylyl cyclase synthesizes the cAMP required for the acrosome reaction. Epac stimulates the exchange of GDP for GTP on Rap1, upstream of a phospholipase C. The Epac-selective cAMP analogue 8-pCPT-2′-O-Me-cAMP induces a phospholipase C-dependent calcium mobilization in human sperm suspensions. In addition, our studies identify a novel connection between cAMP and Rab3A, a secretory granule-associated protein, revealing that the latter functions downstream of soluble adenylyl cyclase/cAMP/Epac but not of Rap1. Challenging sperm with calcium or 8-pCPT-2′-O-Me-cAMP boosts the exchange of GDP for GTP on Rab3A. Recombinant Epac does not release GDP from Rab3A in vitro, suggesting that the Rab3A-GEF activation by cAMP/Epac in vivo is indirect. We propose that Epac sits at a critical point during the exocytotic cascade after which the pathway splits into two limbs, one that assembles the fusion machinery into place and another that elicits intracellular calcium release. PMID:19546222

  2. nRap GEP: a novel neural GDP/GTP exchange protein for rap1 small G protein that interacts with synaptic scaffolding molecule (S-SCAM).

    PubMed

    Ohtsuka, T; Hata, Y; Ide, N; Yasuda, T; Inoue, E; Inoue, T; Mizoguchi, A; Takai, Y

    1999-11-01

    Synaptic scaffolding molecule (S-SCAM) has six PDZ domains through which it interacts with N-methyl-d-aspartate receptors and neuroligin at synaptic junctions. We isolated here a novel S-SCAM-binding protein. This protein has one PDZ, one Ras association, one Ras GDP/GTP exchange protein (Ras GEP) domain, and one C-terminal consensus motif for binding to PDZ domains. We named it nRap GEP (neural Rap GEP). nRap GEP moreover has an incomplete cyclic AMP (cAMP)-binding (CAB) domain. The domain organization of nRap GEP is similar to that of Epac/cAMP-guanine nucleotide exchange factor (GEF) I, except that Epac/cAMP-GEFI has complete CAB and Ras GEP domains but lacks the other two domains and the C-terminal motif. nRap GEP showed GEP activity for Rap1 but did not bind cAMP. nRap GEP was specifically expressed in rat brain. Immunohistochemical analysis revealed that nRap GEP and S-SCAM were localized at synaptic areas of the cerebellum. These results suggest that nRap GEP is a novel neural Rap1-specific GEP which is associated with S-SCAM.

  3. PKA, Rap1, ERK1/2, and p90RSK mediate PGE2 and EP4 signaling in neonatal ventricular myocytes.

    PubMed

    He, Quan; Harding, Pamela; LaPointe, Margot C

    2010-01-01

    We have previously reported that 1) inhibition of cyclooxygenase-2 and PGE(2) production reduces hypertrophy after myocardial infarction in mice and 2) PGE(2) acting through its EP4 receptor causes hypertrophy of neonatal ventricular myocytes (NVMs) via ERK1/2. It is known that EP4 couples to adenylate cyclase, cAMP, and PKA. The present study was designed to determine interactions between the cAMP-PKA pathway and ERK1/2 and to further characterize events downstream of ERK1/2. We hypothesized that PKA and the small GTPase Rap are upstream of ERK1/2 and that 90-kDa ribosomal S6 kinase (p90RSK) is activated downstream. Treatment of NVMs with PGE(2) activated Rap, and this activation was inhibited in part by an EP4 antagonist and PKA inhibition. Transfection of a dominant negative mutant of Rap reduced PGE(2) activation of ERK1/2. PGE(2) activation of p90RSK was also dependent on EP4, PKA, and Rap. We also tested the involvement of Rap, ERK1/2, and p90RSK in PGE(2) regulation of gene expression. PGE(2) stimulation of brain natriuretic peptide promoter activity was blocked by either ERK1/2 inhibition or a dominant negative mutation of p90RSK. PGE(2) stimulation of c-Fos was dependent on EP4, PKA, ERK1/2, and p90RSK, whereas only the latter two kinases were involved in PGE(2) regulation of early growth response-1. Finally, we tested the involvement of EP4-dependent signaling in the NVM growth response and found that the overexpression of EP4 increased NVM cell size. We conclude that EP4-dependent signaling in NVMs in part involves PKA, Rap, ERK1/2, and p90RSK and results in the increased expression of brain natriuretic peptide and c-Fos.

  4. Source localization using recursively applied and projected (RAP) MUSIC

    SciTech Connect

    Mosher, J.C.; Leahy, R.M.

    1998-03-01

    A new method for source localization is described that is based on a modification of the well known multiple signal classification (MUSIC) algorithm. In classical MUSIC, the array manifold vector is projected onto an estimate of the signal subspace, but errors in the estimate can make location of multiple sources difficult. Recursively applied and projected (RAP) MUSIC uses each successively located source to form an intermediate array gain matrix, and projects both the array manifold and the signal subspace estimate into its orthogonal complement. The MUSIC projection is then performed in this reduced subspace. Using the metric of principal angles, the authors describe a general form of the RAP-MUSIC algorithm for the case of diversely polarized sources. Through a uniform linear array simulation, the authors demonstrate the improved Monte Carlo performance of RAP-MUSIC relative to MUSIC and two other sequential subspace methods, S and IES-MUSIC.

  5. Novel Alternative Splice Variants of Mouse Cdk5rap2

    PubMed Central

    Kraemer, Nadine; Issa-Jahns, Lina; Neubert, Gerda; Ravindran, Ethiraj; Mani, Shyamala; Ninnemann, Olaf; Kaindl, Angela M.

    2015-01-01

    Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental disorder characterized by a pronounced reduction of brain volume and intellectual disability. A current model for the microcephaly phenotype invokes a stem cell proliferation and differentiation defect, which has moved the disease into the spotlight of stem cell biology and neurodevelopmental science. Homozygous mutations of the Cyclin-dependent kinase-5 regulatory subunit-associated protein 2 gene CDK5RAP2 are one genetic cause of MCPH. To further characterize the pathomechanism underlying MCPH, we generated a conditional Cdk5rap2 LoxP/hCMV Cre mutant mouse. Further analysis, initiated on account of a lack of a microcephaly phenotype in these mutant mice, revealed the presence of previously unknown splice variants of the Cdk5rap2 gene that are at least in part accountable for the lack of microcephaly in the mice. PMID:26322982

  6. Novel Alternative Splice Variants of Mouse Cdk5rap2.

    PubMed

    Kraemer, Nadine; Issa-Jahns, Lina; Neubert, Gerda; Ravindran, Ethiraj; Mani, Shyamala; Ninnemann, Olaf; Kaindl, Angela M

    2015-01-01

    Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental disorder characterized by a pronounced reduction of brain volume and intellectual disability. A current model for the microcephaly phenotype invokes a stem cell proliferation and differentiation defect, which has moved the disease into the spotlight of stem cell biology and neurodevelopmental science. Homozygous mutations of the Cyclin-dependent kinase-5 regulatory subunit-associated protein 2 gene CDK5RAP2 are one genetic cause of MCPH. To further characterize the pathomechanism underlying MCPH, we generated a conditional Cdk5rap2 LoxP/hCMV Cre mutant mouse. Further analysis, initiated on account of a lack of a microcephaly phenotype in these mutant mice, revealed the presence of previously unknown splice variants of the Cdk5rap2 gene that are at least in part accountable for the lack of microcephaly in the mice. PMID:26322982

  7. Rap and Race: It's Got a Nice Beat, but What about the Message?

    ERIC Educational Resources Information Center

    Sullivan, Rachel E.

    2003-01-01

    Examined racial differences in black and white adolescents' preferences for and interpretations of rap music. Surveys of midwestern U.S. adolescents highlighted limited racial differences in the popularity of rap music. African American youth were more committed to rap and more likely to see it as life affirming. Though both groups had favorable…

  8. 40 CFR 270.200 - How may I renew my RAP if it is expiring?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false How may I renew my RAP if it is expiring? 270.200 Section 270.200 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... renew my RAP if it is expiring? If you wish to renew your expiring RAP, you must follow the process...

  9. RNA antisense purification (RAP) for mapping RNA interactions with chromatin.

    PubMed

    Engreitz, Jesse; Lander, Eric S; Guttman, Mitchell

    2015-01-01

    RNA-centric biochemical purification is a general approach for studying the functions and mechanisms of noncoding RNAs. Here, we describe the experimental procedures for RNA antisense purification (RAP), a method for selective purification of endogenous RNA complexes from cell extracts that enables mapping of RNA interactions with chromatin. In RAP, the user cross-links cells to fix endogenous RNA complexes and purifies these complexes through hybrid capture with biotinylated antisense oligos. DNA loci that interact with the target RNA are identified using high-throughput DNA sequencing.

  10. RAP: RNA-Seq Analysis Pipeline, a new cloud-based NGS web application

    PubMed Central

    2015-01-01

    Background The study of RNA has been dramatically improved by the introduction of Next Generation Sequencing platforms allowing massive and cheap sequencing of selected RNA fractions, also providing information on strand orientation (RNA-Seq). The complexity of transcriptomes and of their regulative pathways make RNA-Seq one of most complex field of NGS applications, addressing several aspects of the expression process (e.g. identification and quantification of expressed genes and transcripts, alternative splicing and polyadenylation, fusion genes and trans-splicing, post-transcriptional events, etc.). Moreover, the huge volume of data generated by NGS platforms introduces unprecedented computational and technological challenges to efficiently analyze and store sequence data and results. Methods In order to provide researchers with an effective and friendly resource for analyzing RNA-Seq data, we present here RAP (RNA-Seq Analysis Pipeline), a cloud computing web application implementing a complete but modular analysis workflow. This pipeline integrates both state-of-the-art bioinformatics tools for RNA-Seq analysis and in-house developed scripts to offer to the user a comprehensive strategy for data analysis. RAP is able to perform quality checks (adopting FastQC and NGS QC Toolkit), identify and quantify expressed genes and transcripts (with Tophat, Cufflinks and HTSeq), detect alternative splicing events (using SpliceTrap) and chimeric transcripts (with ChimeraScan). This pipeline is also able to identify splicing junctions and constitutive or alternative polyadenylation sites (implementing custom analysis modules) and call for statistically significant differences in genes and transcripts expression, splicing pattern and polyadenylation site usage (using Cuffdiff2 and DESeq). Results Through a user friendly web interface, the RAP workflow can be suitably customized by the user and it is automatically executed on our cloud computing environment. This strategy

  11. Bidirectional Synaptic Structural Plasticity after Chronic Cocaine Administration Occurs through Rap1 Small GTPase Signaling.

    PubMed

    Cahill, Michael E; Bagot, Rosemary C; Gancarz, Amy M; Walker, Deena M; Sun, HaoSheng; Wang, Zi-Jun; Heller, Elizabeth A; Feng, Jian; Kennedy, Pamela J; Koo, Ja Wook; Cates, Hannah M; Neve, Rachael L; Shen, Li; Dietz, David M; Nestler, Eric J

    2016-02-01

    Dendritic spines are the sites of most excitatory synapses in the CNS, and opposing alterations in the synaptic structure of medium spiny neurons (MSNs) of the nucleus accumbens (NAc), a primary brain reward region, are seen at early versus late time points after cocaine administration. Here we investigate the time-dependent molecular and biochemical processes that regulate this bidirectional synaptic structural plasticity of NAc MSNs and associated changes in cocaine reward in response to chronic cocaine exposure. Our findings reveal key roles for the bidirectional synaptic expression of the Rap1b small GTPase and an associated local synaptic protein translation network in this process. The transcriptional mechanisms and pathway-specific inputs to NAc that regulate Rap1b expression are also characterized. Collectively, these findings provide a precise mechanism by which nuclear to synaptic interactions induce "metaplasticity" in NAc MSNs, and we reveal the specific effects of this plasticity on reward behavior in a brain circuit-specific manner. PMID:26844834

  12. 40 CFR 270.225 - What must the State or EPA Region report about noncompliance with RAPs?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... report about noncompliance with RAPs? 270.225 Section 270.225 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.225 What must the State or EPA Region report about noncompliance with RAPs? The State or EPA Region must report noncompliance with...

  13. 40 CFR 270.130 - What is the process for approving or denying my application for a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... denying my application for a RAP? 270.130 Section 270.130 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.130 What is the process for approving or denying my application for a RAP? (a) If the Director tentatively finds that your...

  14. 40 CFR 270.150 - How will the Director make a final decision on my RAP application?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... decision on my RAP application? 270.150 Section 270.150 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.150 How will the Director make a final decision on my RAP application? (a) The Director must consider and respond to any significant...

  15. 40 CFR 270.215 - How are time periods in the requirements in this subpart and my RAP computed?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... requirements in this subpart and my RAP computed? 270.215 Section 270.215 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.215 How are time periods in the requirements in this subpart and my RAP computed? (a) Any time period scheduled to begin...

  16. 40 CFR 270.90 - Does my RAP grant me any rights or relieve me of any obligations?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false Does my RAP grant me any rights or... PROGRAM Remedial Action Plans (RAPs) General Information § 270.90 Does my RAP grant me any rights or relieve me of any obligations? The provisions of § 270.4 apply to RAPs. (Note: The provisions of §...

  17. 40 CFR 270.150 - How will the Director make a final decision on my RAP application?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... decision on my RAP application? 270.150 Section 270.150 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.150 How will the Director make a final decision on my RAP application? (a) The Director must consider and respond to any significant...

  18. 40 CFR 270.130 - What is the process for approving or denying my application for a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... denying my application for a RAP? 270.130 Section 270.130 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.130 What is the process for approving or denying my application for a RAP? (a) If the Director tentatively finds that your...

  19. 40 CFR 270.165 - When may I begin physical construction of new units permitted under the RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... of new units permitted under the RAP? 270.165 Section 270.165 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.165 When may I begin physical construction of new units permitted under the RAP? You must not begin physical construction of new...

  20. 40 CFR 270.170 - After my RAP is issued, how may it be modified, revoked and reissued, or terminated?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false After my RAP is issued, how may it be... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.170 After my RAP is issued, how may it be modified, revoked and reissued,...

  1. 40 CFR 270.225 - What must the State or EPA Region report about noncompliance with RAPs?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... report about noncompliance with RAPs? 270.225 Section 270.225 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.225 What must the State or EPA Region report about noncompliance with RAPs? The State or EPA Region must report noncompliance with...

  2. 40 CFR 270.150 - How will the Director make a final decision on my RAP application?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... decision on my RAP application? 270.150 Section 270.150 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.150 How will the Director make a final decision on my RAP application? (a) The Director must consider and respond to any significant...

  3. 40 CFR 270.90 - Does my RAP grant me any rights or relieve me of any obligations?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false Does my RAP grant me any rights or... PROGRAM Remedial Action Plans (RAPs) General Information § 270.90 Does my RAP grant me any rights or relieve me of any obligations? The provisions of § 270.4 apply to RAPs. (Note: The provisions of §...

  4. 40 CFR 270.170 - After my RAP is issued, how may it be modified, revoked and reissued, or terminated?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false After my RAP is issued, how may it be... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.170 After my RAP is issued, how may it be modified, revoked and reissued,...

  5. 40 CFR 270.155 - May the decision to approve or deny my RAP application be administratively appealed?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... RAP application be administratively appealed? 270.155 Section 270.155 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May the decision to approve or deny my RAP application be administratively appealed? (a) Any commenter on the...

  6. 40 CFR 270.155 - May the decision to approve or deny my RAP application be administratively appealed?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... RAP application be administratively appealed? 270.155 Section 270.155 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May the decision to approve or deny my RAP application be administratively appealed? (a) Any commenter on the...

  7. 40 CFR 270.225 - What must the State or EPA Region report about noncompliance with RAPs?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... report about noncompliance with RAPs? 270.225 Section 270.225 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.225 What must the State or EPA Region report about noncompliance with RAPs? The State or EPA Region must report noncompliance with...

  8. 40 CFR 270.155 - May the decision to approve or deny my RAP application be administratively appealed?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... RAP application be administratively appealed? 270.155 Section 270.155 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May the decision to approve or deny my RAP application be administratively appealed? (a) Any commenter on the...

  9. 40 CFR 270.215 - How are time periods in the requirements in this subpart and my RAP computed?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... requirements in this subpart and my RAP computed? 270.215 Section 270.215 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.215 How are time periods in the requirements in this subpart and my RAP computed? (a) Any time period scheduled to begin...

  10. 40 CFR 270.175 - For what reasons may the Director choose to modify my final RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... choose to modify my final RAP? 270.175 Section 270.175 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.175 For what reasons may the Director choose to modify my final RAP? (a) The Director may...

  11. 40 CFR 270.170 - After my RAP is issued, how may it be modified, revoked and reissued, or terminated?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false After my RAP is issued, how may it be... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.170 After my RAP is issued, how may it be modified, revoked and reissued,...

  12. 40 CFR 270.150 - How will the Director make a final decision on my RAP application?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... decision on my RAP application? 270.150 Section 270.150 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.150 How will the Director make a final decision on my RAP application? (a) The Director must consider and respond to any significant...

  13. 40 CFR 270.165 - When may I begin physical construction of new units permitted under the RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... of new units permitted under the RAP? 270.165 Section 270.165 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.165 When may I begin physical construction of new units permitted under the RAP? You must not begin physical construction of new...

  14. 40 CFR 270.225 - What must the State or EPA Region report about noncompliance with RAPs?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... report about noncompliance with RAPs? 270.225 Section 270.225 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.225 What must the State or EPA Region report about noncompliance with RAPs? The State or EPA Region must report noncompliance with...

  15. 40 CFR 270.175 - For what reasons may the Director choose to modify my final RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... choose to modify my final RAP? 270.175 Section 270.175 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.175 For what reasons may the Director choose to modify my final RAP? (a) The Director may...

  16. 40 CFR 270.165 - When may I begin physical construction of new units permitted under the RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... of new units permitted under the RAP? 270.165 Section 270.165 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.165 When may I begin physical construction of new units permitted under the RAP? You must not begin physical construction of new...

  17. 40 CFR 270.130 - What is the process for approving or denying my application for a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... denying my application for a RAP? 270.130 Section 270.130 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.130 What is the process for approving or denying my application for a RAP? (a) If the Director tentatively finds that your...

  18. 40 CFR 270.225 - What must the State or EPA Region report about noncompliance with RAPs?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... report about noncompliance with RAPs? 270.225 Section 270.225 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.225 What must the State or EPA Region report about noncompliance with RAPs? The State or EPA Region must report noncompliance with...

  19. 40 CFR 270.130 - What is the process for approving or denying my application for a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... denying my application for a RAP? 270.130 Section 270.130 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.130 What is the process for approving or denying my application for a RAP? (a) If the Director tentatively finds that your...

  20. 40 CFR 270.215 - How are time periods in the requirements in this subpart and my RAP computed?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... requirements in this subpart and my RAP computed? 270.215 Section 270.215 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.215 How are time periods in the requirements in this subpart and my RAP computed? (a) Any time period scheduled to begin...

  1. 40 CFR 270.165 - When may I begin physical construction of new units permitted under the RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... of new units permitted under the RAP? 270.165 Section 270.165 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.165 When may I begin physical construction of new units permitted under the RAP? You must not begin physical construction of new...

  2. 40 CFR 270.170 - After my RAP is issued, how may it be modified, revoked and reissued, or terminated?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false After my RAP is issued, how may it be... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.170 After my RAP is issued, how may it be modified, revoked and reissued,...

  3. 40 CFR 270.165 - When may I begin physical construction of new units permitted under the RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... of new units permitted under the RAP? 270.165 Section 270.165 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.165 When may I begin physical construction of new units permitted under the RAP? You must not begin physical construction of new...

  4. 40 CFR 270.90 - Does my RAP grant me any rights or relieve me of any obligations?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false Does my RAP grant me any rights or... PROGRAM Remedial Action Plans (RAPs) General Information § 270.90 Does my RAP grant me any rights or relieve me of any obligations? The provisions of § 270.4 apply to RAPs. (Note: The provisions of §...

  5. 40 CFR 270.170 - After my RAP is issued, how may it be modified, revoked and reissued, or terminated?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false After my RAP is issued, how may it be... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.170 After my RAP is issued, how may it be modified, revoked and reissued,...

  6. 40 CFR 270.155 - May the decision to approve or deny my RAP application be administratively appealed?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... RAP application be administratively appealed? 270.155 Section 270.155 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May the decision to approve or deny my RAP application be administratively appealed? (a) Any commenter on the...

  7. 40 CFR 270.150 - How will the Director make a final decision on my RAP application?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... decision on my RAP application? 270.150 Section 270.150 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.150 How will the Director make a final decision on my RAP application? (a) The Director must consider and respond to any significant...

  8. 40 CFR 270.90 - Does my RAP grant me any rights or relieve me of any obligations?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Does my RAP grant me any rights or... PROGRAM Remedial Action Plans (RAPs) General Information § 270.90 Does my RAP grant me any rights or relieve me of any obligations? The provisions of § 270.4 apply to RAPs. (Note: The provisions of §...

  9. 40 CFR 270.90 - Does my RAP grant me any rights or relieve me of any obligations?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false Does my RAP grant me any rights or... PROGRAM Remedial Action Plans (RAPs) General Information § 270.90 Does my RAP grant me any rights or relieve me of any obligations? The provisions of § 270.4 apply to RAPs. (Note: The provisions of §...

  10. 40 CFR 270.215 - How are time periods in the requirements in this subpart and my RAP computed?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... requirements in this subpart and my RAP computed? 270.215 Section 270.215 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.215 How are time periods in the requirements in this subpart and my RAP computed? (a) Any time period scheduled to begin...

  11. 40 CFR 270.130 - What is the process for approving or denying my application for a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... denying my application for a RAP? 270.130 Section 270.130 Protection of Environment ENVIRONMENTAL... PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.130 What is the process for approving or denying my application for a RAP? (a) If the Director tentatively finds that your...

  12. 40 CFR 270.155 - May the decision to approve or deny my RAP application be administratively appealed?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... RAP application be administratively appealed? 270.155 Section 270.155 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May the decision to approve or deny my RAP application be administratively appealed? (a) Any commenter on the...

  13. 40 CFR 270.175 - For what reasons may the Director choose to modify my final RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... choose to modify my final RAP? 270.175 Section 270.175 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.175 For what reasons may the Director choose to modify my final RAP? (a) The Director may...

  14. 40 CFR 270.175 - For what reasons may the Director choose to modify my final RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... choose to modify my final RAP? 270.175 Section 270.175 Protection of Environment ENVIRONMENTAL PROTECTION... PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.175 For what reasons may the Director choose to modify my final RAP? (a) The Director may...

  15. 40 CFR 270.215 - How are time periods in the requirements in this subpart and my RAP computed?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... requirements in this subpart and my RAP computed? 270.215 Section 270.215 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Operating Under Your Rap § 270.215 How are time periods in the requirements in this subpart and my RAP computed? (a) Any time period scheduled to begin...

  16. Paired MEG data set source localization using recursively applied and projected (RAP) MUSIC.

    PubMed

    Ermer, J J; Mosher, J C; Huang, M; Leahy, R M

    2000-09-01

    An important class of experiments in functional brain mapping involves collecting pairs of data corresponding to separate "Task" and "Control" conditions. The data are then analyzed to determine what activity occurs during the Task experiment but not in the Control. Here we describe a new method for processing paired magnetoencephalographic (MEG) data sets using our recursively applied and projected multiple signal classification (RAP-MUSIC) algorithm. In this method the signal subspace of the Task data is projected against the orthogonal complement of the Control data signal subspace to obtain a subspace which describes spatial activity unique to the Task. A RAP-MUSIC localization search is then performed on this projected data to localize the sources which are active in the Task but not in the Control data. In addition to dipolar sources, effective blocking of more complex sources, e.g., multiple synchronously activated dipoles or synchronously activated distributed source activity, is possible since these topographies are well-described by the Control data signal subspace. Unlike previously published methods, the proposed method is shown to be effective in situations where the time series associated with Control and Task activity possess significant cross correlation. The method also allows for straightforward determination of the estimated time series of the localized target sources. A multiepoch MEG simulation and a phantom experiment are presented to demonstrate the ability of this method to successfully identify sources and their time series in the Task data.

  17. Multilingual Codeswitching in Quebec Rap: Poetry, Pragmatics and Performativity

    ERIC Educational Resources Information Center

    Sarkar, Mela; Winer, Lise

    2006-01-01

    Quebec rap lyrics stand out on the world Hip-Hop scene by virtue of the ease and rapidity with which performers in this multilingual, multiethnic youth community codeswitch, frequently among three or more languages or language varieties (usually over a French and/or English base) in the same song. We construct a framework for understanding…

  18. Rap Therapy? An Innovative Approach to Groupwork with Urban Adolescents.

    ERIC Educational Resources Information Center

    DeCarlo, Alonzo

    2001-01-01

    Describes a study in which young, urban African American adolescents with behavior problems participated in weekly group sessions that used rap music to promote the development of appropriate social skills related to morality, identity, judgement, decision making, anger management, impulse control, and crime and punishment. Overall, student…

  19. Escaping Embarrassment: Face-Work in the Rap Cipher

    ERIC Educational Resources Information Center

    Lee, Jooyoung

    2009-01-01

    How do individuals escape embarrassing moments in interaction? Drawing from ethnographic fieldwork, in-depth interviews, and video recordings of weekly street corner ciphers (impromptu rap sessions), this paper expands Goffman's theory of defensive and protective face-work. The findings reveal formulaic and indirect dimensions of face-work. First,…

  20. Hybrid Texts: Fifth Graders, Rap Music, and Writing

    ERIC Educational Resources Information Center

    Christianakis, Mary

    2011-01-01

    Consistent with a sociocritical frame and the analytic tools of hybridity theory, this article explicates how urban fifth-grade children made language hybrids using rap and poetry to participate in classroom literacy. Ethnographic data from a yearlong study illustrate two key findings. First, standards-based and canon-driven writing models…

  1. Rap Music by Black Male Artists: A Psychotheological Interpretation.

    ERIC Educational Resources Information Center

    Pressley, Arthur

    1992-01-01

    Provides a psychotheological interpretation of rap music by African-American male artists and of its audience, examining the music and its social context. Common themes include despair over acute psychosocial and physical needs, intensity and violence as a means of personal integration, ontological insecurity, and desire for transformation and…

  2. Fresh Out of School: Rap Music's Discursive Battle with Education

    ERIC Educational Resources Information Center

    Au, Wayne

    2005-01-01

    The rap music lyrics were analyzed to flush out the hip-hop culture's perspective on the education of African American (AA) youth. It was found that there is a need for the implementation of more culturally relevant curricula in schools, which benefits the students to understand hip-hop culture.

  3. Structure of an LDLR-RAP Complex Reveals a General Mode for Ligand Recognition by Lipoprotein Receptors

    SciTech Connect

    Fisher,C.; Beglova, N.; Blacklow, s.

    2006-01-01

    Proteins of the low-density lipoprotein receptor (LDLR) family are remarkable in their ability to bind an extremely diverse range of protein and lipoprotein ligands, yet the basis for ligand recognition is poorly understood. Here, we report the 1.26 Angstroms X-ray structure of a complex between a two-module region of the ligand binding domain of the LDLR and the third domain of RAP, an escort protein for LDLR family members. The RAP domain forms a three-helix bundle with two docking sites, one for each LDLR module. The mode of recognition at each site is virtually identical: three conserved, calcium-coordinating acidic residues from each LDLR module encircle a lysine side chain protruding from the second helix of RAP. This metal-dependent mode of electrostatic recognition, together with avidity effects resulting from the use of multiple sites, represents a general binding strategy likely to apply in the binding of other basic ligands to LDLR family proteins.

  4. Biosynthesis of pipecolic acid by RapL, a lysine cyclodeaminase encoded in the rapamycin gene cluster.

    PubMed

    Gatto, Gregory J; Boyne, Michael T; Kelleher, Neil L; Walsh, Christopher T

    2006-03-22

    Rapamycin, FK506, and FK520 are immunosuppressant macrolactone natural products comprised of predominantly polyketide-based core structures. A single nonproteinogenic pipecolic acid residue is installed into the scaffold by a nonribosomal peptide synthetase that also performs the subsequent macrocyclization step at the carbonyl group of this amino acid. It has been assumed that pipecolic acid is generated from lysine by the cyclodeaminases RapL/FkbL. Herein we report the heterologous overexpression and purification of RapL and validate its ability to convert L-lysine to L-pipecolic acid by a cyclodeamination reaction that involves redox catalysis. RapL also accepts L-ornithine as a substrate, albeit with a significantly reduced catalytic efficiency. Turnover is presumed to encompass a reversible oxidation at the alpha-amine, internal cyclization, and subsequent re-reduction of the cyclic delta1-piperideine-2-carboxylate intermediate. As isolated, RapL has about 0.17 equiv of tightly bound NAD+, suggesting that the enzyme is incompletely loaded when overproduced in E. coli. In the presence of exogenous NAD+, the initial rate is elevated 8-fold with a Km of 2.3 microM for the cofactor, consistent with some release and rebinding of NAD+ during catalytic cycles. Through the use of isotopically labeled substrates, we have confirmed mechanistic details of the cyclodeaminase reaction, including loss of the alpha-amine and retention of the hydrogen atom at the alpha-carbon. In addition to the characterization of a critical enzyme in the biosynthesis of a medically important class of natural products, this work represents the first in vitro characterization of a lysine cyclodeaminase, a member of a unique group of enzymes which utilize the nicotinamide cofactor in a catalytic manner. PMID:16536560

  5. 40 CFR 270.220 - How may I transfer my RAP to a new owner or operator?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false How may I transfer my RAP to a new... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.220 How may I transfer my RAP to a new owner or operator? (a) If you wish to transfer your RAP to a new owner or operator, you must follow the...

  6. 40 CFR 270.220 - How may I transfer my RAP to a new owner or operator?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false How may I transfer my RAP to a new... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.220 How may I transfer my RAP to a new owner or operator? (a) If you wish to transfer your RAP to a new owner or operator, you must follow the...

  7. 40 CFR 270.220 - How may I transfer my RAP to a new owner or operator?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false How may I transfer my RAP to a new... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.220 How may I transfer my RAP to a new owner or operator? (a) If you wish to transfer your RAP to a new owner or operator, you must follow the...

  8. 40 CFR 270.220 - How may I transfer my RAP to a new owner or operator?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false How may I transfer my RAP to a new... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.220 How may I transfer my RAP to a new owner or operator? (a) If you wish to transfer your RAP to a new owner or operator, you must follow the...

  9. 40 CFR 270.220 - How may I transfer my RAP to a new owner or operator?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false How may I transfer my RAP to a new... Remedial Action Plans (RAPs) Operating Under Your Rap § 270.220 How may I transfer my RAP to a new owner or operator? (a) If you wish to transfer your RAP to a new owner or operator, you must follow the...

  10. 40 CFR 270.185 - For what reasons may the Director choose to terminate my final RAP, or deny my renewal application?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... choose to terminate my final RAP, or deny my renewal application? 270.185 Section 270.185 Protection of... PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified... final RAP, or deny my renewal application? The Director may terminate your final RAP on his...

  11. 40 CFR 270.185 - For what reasons may the Director choose to terminate my final RAP, or deny my renewal application?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... choose to terminate my final RAP, or deny my renewal application? 270.185 Section 270.185 Protection of... PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified... final RAP, or deny my renewal application? The Director may terminate your final RAP on his...

  12. 40 CFR 270.185 - For what reasons may the Director choose to terminate my final RAP, or deny my renewal application?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... choose to terminate my final RAP, or deny my renewal application? 270.185 Section 270.185 Protection of... PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified... final RAP, or deny my renewal application? The Director may terminate your final RAP on his...

  13. 40 CFR 270.185 - For what reasons may the Director choose to terminate my final RAP, or deny my renewal application?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... choose to terminate my final RAP, or deny my renewal application? 270.185 Section 270.185 Protection of... PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified... final RAP, or deny my renewal application? The Director may terminate your final RAP on his...

  14. 40 CFR 270.185 - For what reasons may the Director choose to terminate my final RAP, or deny my renewal application?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... choose to terminate my final RAP, or deny my renewal application? 270.185 Section 270.185 Protection of... PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified... final RAP, or deny my renewal application? The Director may terminate your final RAP on his...

  15. Cloning and characterization of Rap GTPase from the Chinese white shrimp Fenneropenaeus chinensis.

    PubMed

    Ren, Qian; Zhou, Jing; Jia, Yu-Ping; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2012-01-01

    Ras-related protein Rap GTPase has been implicated in cell adhesion, cell proliferation, and cell junction formation. The first shrimp Rap cDNA (FcRap) was recently identified from the Chinese white shrimp Fenneropenaeus chinensis. The full length of FcRap is 1013 bp, with a 561 bp open reading frame that encodes a 186 amino acid protein. FcRap has a calculated molecular mass of 20.90 kDa and pI of 6.37. Phylogenetic analysis shows that FcRap and other Rap proteins are clustered into one group. Results from the quantitative real-time polymerase chain reaction show that FcRap could be detected mainly in the hemocytes, hepatopancreas, stomach, and gills, whereas a relatively lower expression level could be detected in the heart and intestines. FcRap in the hemocytes was upregulated 2h post Vibrio challenge, and it was upregulated 2h post white spot syndrome virus (WSSV) challenge, and peaked at 6h before it declined at 12h. No variation in the FcRap transcript was observed in the gills under the Vibrio challenge, but it was initially downregulated 2h post WSSV challenge, and then it was upregulated and peaked at 6h before it was eventually went down at 12h. The rFcRap protein was successfully expressed in Escherichia coli BL21DE3. The pull-down analysis showed that rFcRap protein could interact with VP28, an envelope protein of WSSV. The probable roles of Rap GTPase in shrimp innate immunity are presented for the first time.

  16. The RAP1GA1 locus for human Rap1-GTPase activating protein 1 maps to chromosome 1p36.1-->p35.

    PubMed

    Weiss, J; Rubinfeld, B; Polakis, P G; McCormick, F; Cavenee, W K; Arden, K C

    1994-01-01

    Using a panel of somatic cell hybrids we have mapped the locus for Rap1-GTPase activating protein 1 (RAP1GA1) to human chromosome 1. Fluorescence in situ hybridization experiments independently confirmed the chromosomal localization and refined it to 1p36.1-->p35.

  17. “Everybody Gotta Have a Dream”: Rap-centered Aspirations among Young Black Males Involved in Rap Music Production – A Qualitative Study

    PubMed Central

    Foster, B. Brian

    2015-01-01

    Youth express diverse desires for their educational and occupational futures. Sometimes these aspirations are directed towards somewhat unconventional careers such as rapping and other types of involvement in rap music production. Although many studies have examined traditional educational and occupational aspirations, less is known about the factors that give rise to rap-centered aspirations and how individuals pursue them, particularly as they transition to early adulthood. Drawing on 54 semi- and unstructured interviews with 29 black young men involved in rap music production, I find that rap-centered aspirations are shaped by a range of factors, most notably feedback regarding one’s rap skills, access to recording and production equipment, and the financial means to maintain involvement in rap music production while also ensuring personal and family economic stability. The young men in the study attached different meanings to their aspirations and sometimes recast their motivations for participating in rap music production in response to various social and economic factors. PMID:26005703

  18. [Activation of autophagy pathway in hippocampus and deterioration of learning and memory ability by intermittent hypoxia in rats after cerebral ischemia].

    PubMed

    Guo, Xiangfei; Zhao, Yaning; Li, Jianmin; Liu, Wenqian; Chen, Changxiang

    2016-09-01

    Objective To investigate the effects of different duration of intermittent hypoxia on the autophagy pathway in the hippocampus and the learning and memory ability after cerebral ischemia in rats. Methods 100 male Wistar rats were randomly divided into sham operation (SO) group, ischemia/reperfusion (I/R) group, intermittent hypoxia for 7 days combined with ischemia/reperfusion (IH7-I/R) group, intermittent hypoxia for 14 days combined with ischemia/reperfusion (IH14-I/R) group, intermittent hypoxia for 21 days combined with ischemia/reperfusion (IH21-I/R) group, n =20 in each group. The rats in IH7-I/R group, IH14-I/R group and IH21-I/R group were respectively subjected to intermittent hypoxia for 7, 14 and 21 days prior to I/R modeling by improved Pulsinelli four-vessel occlusion (4-VO). The morphological changes of nerve cells in the hippocampus of rat brain were detected by HE staining; the levels of mammalian target of rapamycin (mTOR) and beclin 1 mRNA in the hippocampus were determined by quantitative real-time PCR; the distribution of mTOR and beclin 1 in the hippocampus was observed by immunohistochemistry; the learning and memory ability of rats was assessed by the Morris water maze test. Results Compared with the SO group, the never cell morphology was damaged, the number of survival neurons in the hippocampus was reduced, the expressions of mTOR and beclin 1 in the hippocampus were strengthened, and the learning and memory ability declined in the I/R group. Compared with the I/R group, the never cell morphology was damaged seriously, the number of survival neurons in the hippocampus decreased, the expressions of mTOR and beclin 1 in the hippocampus increased, and the learning and memory ability dropped in the intermittent hypoxia groups. What's more, the above changes were dependent on the duration of intermittent hypoxia. Conclusion Intermittent hypoxia aggravates the dysfunction of learning and memory after cerebral ischemia and the damages increase

  19. The signaling module cAMP/Epac/Rap1/PLCε/IP3 mobilizes acrosomal calcium during sperm exocytosis.

    PubMed

    Lucchesi, Ornella; Ruete, María C; Bustos, Matías A; Quevedo, María F; Tomes, Claudia N

    2016-04-01

    Exocytosis of the sperm's single secretory granule, or acrosome, is a regulated exocytosis triggered by components of the egg's investments. In addition to external calcium, sperm exocytosis (termed the acrosome reaction) requires cAMP synthesized endogenously and calcium mobilized from the acrosome through IP3-sensitive channels. The relevant cAMP target is Epac. In the first part of this paper, we present a novel tool (the TAT-cAMP sponge) to investigate cAMP-related signaling pathways in response to progesterone as acrosome reaction trigger. The TAT-cAMP sponge consists of the cAMP-binding sites of protein kinase A regulatory subunit RIβ fused to the protein transduction domain TAT of the human immunodeficiency virus-1. The sponge permeated into sperm, sequestered endogenous cAMP, and blocked exocytosis. Progesterone increased the population of sperm with Rap1-GTP, Rab3-GTP, and Rab27-GTP in the acrosomal region; pretreatment with the TAT-cAMP sponge prevented the activation of all three GTPases. In the second part of this manuscript, we show that phospholipase Cε (PLCε) is required for the acrosome reaction downstream of Rap1 and upstream of intra-acrosomal calcium mobilization. Last, we present direct evidence that cAMP, Epac, Rap1, and PLCε are necessary for calcium mobilization from sperm's secretory granule. In summary, we describe here a pathway that connects cAMP to calcium mobilization from the acrosome during sperm exocytosis. Never before had direct evidence for each step of the cascade been put together in the same study.

  20. What's the hype about CDK5RAP2?

    PubMed

    Kraemer, Nadine; Issa, Lina; Hauck, Stefanie C R; Mani, Shyamala; Ninnemann, Olaf; Kaindl, Angela M

    2011-05-01

    Cyclin dependent kinase 5 regulatory subunit-associated protein 2 (CDK5RAP2) has gained attention in the last years following the discovery, in 2005, that recessive mutations cause primary autosomal recessive microcephaly. This disease is seen as an isolated developmental defect of the brain, particularly of the cerebral cortex, and was thus historically also referred to as microcephalia vera. Unraveling the pathomechanisms leading to this human disease is fascinating scientists because it can convey insight into basic mechanisms of physiologic brain development (particularly of cortex formation). It also finds itself in the spotlight because of its implication in trends in mammalian evolution with a massive increase in the size of the cerebral cortex in primates. Here, we provide a timely overview of the current knowledge on the function of CDK5RAP2 and mechanisms that might lead to disease in humans when the function of this protein is disturbed. PMID:21327915

  1. Dynamic recruitment of CDK5RAP2 to centrosomes requires its association with dynein.

    PubMed

    Jia, Yue; Fong, Ka-Wing; Choi, Yuk-Kwan; See, Siu-San; Qi, Robert Z

    2013-01-01

    CDK5RAP2 is a centrosomal protein known to be involved in the regulation of the γ-tubulin ring complex and thus the organization of microtubule arrays. However, the mechanism by which CDK5RAP2 is itself recruited to centrosomes is poorly understood. We report here that CDK5RAP2 displays highly dynamic attachment to centrosomes in a microtubule-dependent manner. CDK5RAP2 associates with the retrograde transporter dynein-dynactin and contains a sequence motif that binds to dynein light chain 8. Significantly, disruption of cellular dynein-dynactin function reduces the centrosomal level of CDK5RAP2. These results reveal a key role of the dynein-dynactin complex in the dynamic recruitment of CDK5RAP2 to centrosomes. PMID:23874654

  2. Assisted annotation of medical free text using RapTAT

    PubMed Central

    Gobbel, Glenn T; Garvin, Jennifer; Reeves, Ruth; Cronin, Robert M; Heavirland, Julia; Williams, Jenifer; Weaver, Allison; Jayaramaraja, Shrimalini; Giuse, Dario; Speroff, Theodore; Brown, Steven H; Xu, Hua; Matheny, Michael E

    2014-01-01

    Objective To determine whether assisted annotation using interactive training can reduce the time required to annotate a clinical document corpus without introducing bias. Materials and methods A tool, RapTAT, was designed to assist annotation by iteratively pre-annotating probable phrases of interest within a document, presenting the annotations to a reviewer for correction, and then using the corrected annotations for further machine learning-based training before pre-annotating subsequent documents. Annotators reviewed 404 clinical notes either manually or using RapTAT assistance for concepts related to quality of care during heart failure treatment. Notes were divided into 20 batches of 19–21 documents for iterative annotation and training. Results The number of correct RapTAT pre-annotations increased significantly and annotation time per batch decreased by ∼50% over the course of annotation. Annotation rate increased from batch to batch for assisted but not manual reviewers. Pre-annotation F-measure increased from 0.5 to 0.6 to >0.80 (relative to both assisted reviewer and reference annotations) over the first three batches and more slowly thereafter. Overall inter-annotator agreement was significantly higher between RapTAT-assisted reviewers (0.89) than between manual reviewers (0.85). Discussion The tool reduced workload by decreasing the number of annotations needing to be added and helping reviewers to annotate at an increased rate. Agreement between the pre-annotations and reference standard, and agreement between the pre-annotations and assisted annotations, were similar throughout the annotation process, which suggests that pre-annotation did not introduce bias. Conclusions Pre-annotations generated by a tool capable of interactive training can reduce the time required to create an annotated document corpus by up to 50%. PMID:24431336

  3. Sequence and organization of the rhoptry-associated-protein-1 (rap-1) locus for the sheep hemoprotozoan Babesia sp. BQ1 Lintan (B. motasi phylogenetic group).

    PubMed

    Niu, Qingli; Bonsergent, Claire; Guan, Guiquan; Yin, Hong; Malandrin, Laurence

    2013-11-15

    Babesiosis is a frequent infection of animals worldwide by tick borne pathogen Babesia, and several species are responsible for ovine babesiosis. Recently, several Babesia motasi-like isolates were described in sheep in China. In this study, we sequenced the multigenic rap-1 gene locus of one of these isolates, Babesia sp. BQ1 Lintan. The RAP-1 proteins are involved in the process of red blood cells invasion and thus represent a potential target for vaccine development. A complex composition and organization of the rap-1 locus was discovered with: (1) the presence of 3 different types of rap-1 sequences (rap-1a, rap-1b and rap-1c); (2) the presence of multiple copies of rap-1a and rap-1b; (3) polymorphism among the rap-1a copies, with two classes (named rap-1a61 and rap-1a67) having a similarity of 95.7%, each class represented by two close variants; (4) polymorphism between rap-1a61-1 and rap-1a61-2 limited to three nucleotide positions; (5) a difference of eight nucleotides between rap-1a67-1 and rap-1a67-2 from position 1270 to the putative stop site of rap-1a67-1 which might produce two putative proteins of slightly different sizes; (6) the ratio of rap-1a copies corresponding to one rap-1a67, one rap-1a61-1 and one rap-1a61-2; (7) the presence of three different intergenic regions separating rap-1a, rap-1b and rap-1c; (8) interspacing of the rap-1a copies with rap-1b copies; and (9) the terminal position of rap-1c in the locus. A 31kb locus composed of 6 rap-1a sequences interspaced with 5 rap-1b sequences and with a terminal rap-1c copy was hypothesized. A strikingly similar sequence composition (rap-1a, rap-1b and rap-1c), as well as strong gene identities and similar locus organization with B. bigemina were found and highlight the conservation of synteny at this locus in this phylogenetic clade.

  4. Rapid Fabrication of Lightweight SiC Optics using Reactive Atom Plasma (RAP) Processing

    NASA Technical Reports Server (NTRS)

    Fiske, Peter S.

    2006-01-01

    Reactive Atom Plasma (RAP) processing is a non-contact, plasma-based processing technology that can be used to generate damage-free optical surfaces. We have developed tools and processes using RAP that allow us to shape extremely lightweight mirror Surfaces made from extremely hard-to-machine materials (e.g. SiC). We will describe our latest results using RAP in combination with other technologies to produce finished lightweight SiC mirrors and also discuss applications for RAP in the rapid fabrication of mirror segments for reflective and grazing incidence telescopes.

  5. YBX1 is a modulator of MIA/CD-RAP-dependent chondrogenesis.

    PubMed

    Schmid, Rainer; Meyer, Katharina; Spang, Rainer; Schittek, Birgit; Bosserhoff, Anja Katrin

    2013-01-01

    MIA/CD-RAP is a small, secreted protein involved in cartilage differentiation and melanoma progression. We recently revealed that p54(nrb) acts as a mediator of MIA/CD-RAP action to promote chondrogenesis and the progression of malignant melanoma. As the molecular mechanism of MIA/CD-RAP action in cartilage has not been defined in detail until now, we aimed to understand the regulation of p54(nrb) transcription in chondrogenesis. We concentrated on the previously described MIA/CD-RAP-dependent regulatory region in the p54(nrb) promoter and characterized the transcriptional regulation of p54(nrb) by MIA/CD-RAP in cartilage. A series of truncated p54(nrb) promoter constructs and mutagenesis analysis revealed that the transcription factor YBX1, which has not been investigated in chondrogenesis thus far, is the mediator of MIA/CD-RAP dependent activation of p54(nrb) transcription. A systematic analysis of genes carrying this binding site in their promoter region revealed further potential MIA/CD-RAP-regulated genes that have been implicated in cartilage differentiation. In summary, we described the effects of MIA/CD-RAP on transcriptional regulation in chondrocytes. Understanding the regulation of p54(nrb) via YBX1 contributes to the understanding of chondrogenesis. Uncovering new downstream effectors that function via the activation of YBX1 supports the important role of MIA/CD-RAP in these processes.

  6. Translation of the Risk Avoidance Partnership (RAP) for Implementation in Outpatient Drug Treatment Clinics.

    PubMed

    Weeks, Margaret R; Kostick, Kristin; Li, Jianghong; Dunn, Jennifer; McLaughlin, Paul; Richmond, Phil; Choudhury, Shonali; Obidoa, Chinekwu; Mosher, Heather; Martinez, Maria

    2015-01-01

    Scientific literature increasingly calls for studies to translate evidence-based interventions into real-world contexts balancing fidelity to the original design and fit to the new setting. The Risk Avoidance Partnership (RAP) is a health promotion intervention originally designed to train active drug users to become Peer Health Advocates. A theoretically driven approach was used to adapt RAP to fit implementation in outpatient methadone treatment clinics and pilot it with clinic patients. Ethnographic observations and process tracking documented the RAP translation and pilot experience, and clinic and community characteristics relevant to program implementation. Clinic administrators, staff, and patients were interviewed on their values, capacities, interest in RAP, perceived challenges of implementing RAP in drug treatment clinics, and experiences during the pilot. Findings indicated that RAP core components can be met when implemented in these settings and RAP can fit with the goals, interests, and other programs of the clinic. Balancing fidelity and fit requires recognition of the mutual impacts RAP and the clinic have on each other, which generate new interactions among staff and require ongoing specification of RAP to keep abreast of clinic and community changes. Collaboration of multiple stakeholders significantly benefited translation and pilot processes.

  7. Inhibition of the mevalonate pathway and activation of p38 MAP kinase are independently regulated by nitrogen-containing bisphosphonates in breast cancer cells.

    PubMed

    Merrell, Melinda A; Wakchoure, Savita; Lehenkari, Petri P; Harris, Kevin W; Selander, Katri S

    2007-09-10

    Bisphosphonates are widely used inhibitors of bone resorption. They also inhibit the growth of various cancer cells in vitro, but the clinical significance of this effect is unclear. The cancer growth inhibitory effects of nitrogen-containing bisphosphonates, (i.e. zoledronate) have been attributed to their ability to inhibit the mevalonate pathway. We have shown that bisphosphonates also induce p38 activation, which signals resistance against the drug-induced growth inhibition through an unknown mechanism. We show here that zoledronate induces a G1/S cell cycle arrest in human MDA-MB-231 breast cancer cells. Furthermore, p38 inhibitor augments bisphosphonate-induced growth inhibition by inducing an additional G2-phase cell cycle arrest. We also show that the nitrogen-containing bisphosphonate-induced effects on p38 phosphorylation occur before accumulation of unprenylated Rap1A or Rac1 activation. Geranylgeranyl pyrophosphate, an end-product of the mevalonate pathway, reversed the accumulation of unprenylated Rap1A but not phosphorylation of p38. Geranylgeranyl pyrophosphate also reversed n-BP induced growth inhibition, but the completeness of this reversal was nitrogen-containing bisphosphonate concentration dependent. Also mevastatin induced the accumulation of unprenylated Rap1A, but it did not induce p38 phosphorylation. In conclusion, our results suggest that in addition to the previously reported effects on apoptosis, nitrogen-containing bisphosphonates also inhibit the growth of MDA-MB-231 breast cancer cells by inducing G1/S cell cycle arrest. The bisphosphonate-induced p38 activation signals for resistance against these drugs, by promoting progression through the G2/M-checkpoint. Of these pathways only growth inhibition is mediated via inhibition of the mevalonate pathway in MDA-MB-231 cells. Combining p38 inhibitors with bisphosphonates may result in increased anti-cancer efficacy.

  8. Rap1 GTPase Activation and Barrier Enhancement in RPE Inhibits Choroidal Neovascularization In Vivo

    PubMed Central

    McCloskey, Manabu; Wang, Haibo; Quilliam, Lawrence A.; Chrzanowska-Wodnicka, Magdalena; Hartnett, M. Elizabeth

    2013-01-01

    Loss of barrier integrity precedes the development of pathologies such as metastasis, inflammatory disorders, and blood-retinal barrier breakdown present in neovascular age-related macular degeneration. Rap1 GTPase is involved in regulating both endothelial and epithelial cell junctions; the specific role of Rap1A vs. Rap1B isoforms is less clear. Compromise of retinal pigment epithelium barrier function is a contributing factor to the development of AMD. We utilized shRNA of Rap1 isoforms in cultured human retinal pigment epithelial cells, along with knockout mouse models to test the role of Rap1 on promoting RPE barrier properties, with emphasis on the dynamic junctional regulation that is triggered when the adhesion between cells is challenged. In vitro, Rap1A shRNA reduced steady-state barrier integrity, whereas Rap1B shRNA affected dynamic junctional responses. In a laser-induced choroidal neovascularization (CNV) model of macular degeneration, Rap1b−/− mice exhibited larger CNV volumes compared to wild-type or Rap1a−/−. In vivo, intravitreal injection of a cAMP analog (8CPT-2′-O-Me-cAMP) that is a known Rap1 activator significantly reduced laser-induced CNV volume, which correlated with the inhibition of CEC transmigration across 8CPT-2′O-Me-cAMP-treated RPE monolayers in vitro. Rap1 activation by 8CPT-2′-O-Me-cAMP treatment increased recruitment of junctional proteins and F-actin to cell-cell contacts, increasing both the linearity of junctions in vitro and in cells surrounding laser-induced lesions in vivo. We conclude that in vitro, Rap1A may be important for steady state barrier integrity, while Rap1B is involved more in dynamic junctional responses such as resistance to junctional disassembly induced by EGTA and reassembly of cell junctions following disruption. Furthermore, activation of Rap1 in vivo inhibited development of choroidal neovascular lesions in a laser-injury model. Our data suggest that targeting Rap1 isoforms in vivo with 8

  9. Conformational change-induced repeat domain expansion regulates Rap phosphatase quorum-sensing signal receptors.

    PubMed

    Parashar, Vijay; Jeffrey, Philip D; Neiditch, Matthew B

    2013-01-01

    The large family of Gram-positive quorum-sensing receptors known as the RNPP proteins consists of receptors homologous to the Rap, NprR, PlcR, and PrgX proteins that are regulated by imported oligopeptide autoinducers. Rap proteins are phosphatases and transcriptional anti-activators, and NprR, PlcR, and PrgX proteins are DNA binding transcription factors. Despite their obvious importance, the mechanistic basis of oligopeptide receptor regulation is largely unknown. Here, we report the X-ray crystal structure of the Bacillus subtilis quorum-sensing receptor RapJ in complex with the centrally important oligopeptide autoinducer competence and sporulation factor (CSF, also termed PhrC), a member of the Phr family of quorum-sensing signals. Furthermore, we present the crystal structure of RapI. Comparison of the RapJ-PhrC, RapI, RapH-Spo0F, and RapF-ComA(C) crystal structures reveals the mechanistic basis of Phr activity. More specifically, when complexed with target proteins, Rap proteins consist of a C-terminal tetratricopeptide repeat (TPR) domain connected by a flexible helix-containing linker to an N-terminal 3-helix bundle. In the absence of a target protein or regulatory peptide, the Rap protein 3-helix bundle adopts different conformations. However, in the peptide-bound conformation, the Rap protein N-terminal 3-helix bundle and linker undergo a radical conformational change, form TPR-like folds, and merge with the existing C-terminal TPR domain. To our knowledge, this is the first example of conformational change-induced repeat domain expansion. Furthermore, upon Phr binding, the entire Rap protein is compressed along the TPR superhelical axis, generating new intramolecular contacts that lock the Rap protein in an inactive state. The fact that Rap proteins are conformationally flexible is surprising considering that it is accepted dogma that TPR proteins do not undergo large conformational changes. Repeat proteins are widely used as scaffolds for the

  10. Reelin, Rap1 and N-cadherin orient the migration of multipolar neurons in the developing neocortex.

    PubMed

    Jossin, Yves; Cooper, Jonathan A

    2011-06-01

    Projection neurons migrate from the ventricular zone to the neocortical plate during the development of the mouse brain. Their overall movement is radial, but they become multipolar and move nonradially in the intermediate zone. Here we show that Reelin, the Rap1 GTPase and N-cadherin (NCad) are important for multipolar neurons to polarize their migration toward the cortical plate. Inhibition and rescue experiments indicated that Reelin regulates migration through Rap1 and Akt, and that the Rap1-regulated GTPases RalA, RalB, Rac1 and Cdc42 are also involved. We found that Rap1 regulated the plasma membrane localization of NCad and NCad rescued radial polarization when Rap1 was inhibited. However, inhibition of Rap1 or NCad had little effect on glia-dependent locomotion. We propose a multistep mechanism in which Reelin activates Rap1, Rap1 upregulates NCad, and NCad is needed to orient cell migration.

  11. Rap1 GTPase is required for mouse lens epithelial maintenance and morphogenesis

    PubMed Central

    Maddala, Rupalatha; Nagendran, Tharkika; Lang, Richard A.; Morozov, Alexei; Rao, Ponugoti V.

    2015-01-01

    Rap1, a Ras-like small GTPase, plays a crucial role in cell-matrix adhesive interactions, cell-cell junction formation, cell polarity and migration. The role of Rap1 in vertebrate organ development and tissue architecture, however, remains elusive. We addressed this question in a mouse lens model system using a conditional gene targeting approach. While individual germline deficiency of either Rap1a or Rap1b did not cause overt defects in mouse lens, conditional double deficiency (Rap1 cKO) prior to lens placode formation led to an ocular phenotype including microphthalmia and lens opacification in embryonic mice. The embryonic Rap1 cKO mouse lens exhibited striking defects including loss of E-cadherin- and ZO-1-based cell-cell junctions, disruption of paxillin and β1-integrin-based cell adhesive interactions along with abnormalities in cell shape and apical-basal polarity of epithelium. These epithelial changes were accompanied by increased levels of α-smooth muscle actin, vimentin and N-cadherin, and expression of transcriptional suppressors of E-cadherin (Snai1, Slug and Zeb2), and a mesenchymal metabolic protein (Dihydropyrimidine dehydrogenase). Additionally, while lens differentiation was not overtly affected, increased apoptosis and dysregulated cell cycle progression were noted in epithelium and fibers in Rap1 cKO mice. Collectively these observations uncover a requirement for Rap1 in maintenance of lens epithelial phenotype and morphogenesis. PMID:26212757

  12. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... remediation wastes in a manner that requires a RCRA permit under § 270.1, you must either obtain: (1) A RCRA... an already permitted RCRA facility. You must have these RAPs approved as a modification to your... required under § 270.110. When your permit is modified the RAP becomes part of the RCRA permit....

  13. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... remediation wastes in a manner that requires a RCRA permit under § 270.1, you must either obtain: (1) A RCRA... an already permitted RCRA facility. You must have these RAPs approved as a modification to your... required under § 270.110. When your permit is modified the RAP becomes part of the RCRA permit....

  14. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... remediation wastes in a manner that requires a RCRA permit under § 270.1, you must either obtain: (1) A RCRA... an already permitted RCRA facility. You must have these RAPs approved as a modification to your... required under § 270.110. When your permit is modified the RAP becomes part of the RCRA permit....

  15. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... remediation wastes in a manner that requires a RCRA permit under § 270.1, you must either obtain: (1) A RCRA... an already permitted RCRA facility. You must have these RAPs approved as a modification to your... required under § 270.110. When your permit is modified the RAP becomes part of the RCRA permit....

  16. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... remediation wastes in a manner that requires a RCRA permit under § 270.1, you must either obtain: (1) A RCRA... an already permitted RCRA facility. You must have these RAPs approved as a modification to your... required under § 270.110. When your permit is modified the RAP becomes part of the RCRA permit....

  17. Structural and Functional Implication of RAP80 ΔGlu81 Mutation

    PubMed Central

    Vikrant; Kumar, Rajan; Yadav, Lumbini R.; Nakhwa, Pallavi; Waghmare, Sanjeev K.; Goyal, Peyush; Varma, Ashok K.

    2013-01-01

    Receptor Associated Protein 80 (RAP80) is a member of RAP80-BRCA1-CCDC98 complex family and helps in its recruitment to the DNA damage site for effective homologous recombination repair. It encompasses two tandem UIMs (UIM1 and UIM2) motif at its N-terminus, which interact with K-63 linked polyubiquitin chain(s) on H2AX and thereby assemble the RAP80-BRCA1 complex at the damage site. Nevertheless, how RAP80 helps in the structural integrity of BRCA1 complex is still elusive. Considering the role of RAP80 in the recruitment of BRCA1 complex at the DNA damage site, we attempted to explore the molecular mechanism associated with RAP80 and mutation that causes chromosomal aberrations due to its loss of function. There is a significant loss in structural characteristics of RAP80 ΔE81, which impairs its binding affinity with the polyubiquitin chain. This leads to the defective recruitment of RAP80 and BRCA1 complex at the DNA damage site. The results presented here are very useful in understanding the cause of various repair defects (chromosomal aberration) that arise due to this mutation. Comparative study of wild type and ΔE81 could be helpful in designing the small molecules that can potentially compensate the deleterious effect(s) of ΔE81 and hence useful for therapeutic application. PMID:24039796

  18. Affiliation and Alienation: Hip-Hop, Rap, and Urban Science Education

    ERIC Educational Resources Information Center

    Emdin, Christopher

    2010-01-01

    The critiques of rap artists and other participants in hip-hop culture provide data for teachers and researchers to investigate the attitudes of US urban youth towards schooling. This study explores the complex relationships between hip-hop and science education by examining how rap lyrics project beliefs about schooling, the relevance of existing…

  19. Whats the Rap about Ecstasy? Popular Music Lyrics and Drug Trends among American Youth

    ERIC Educational Resources Information Center

    Diamond, Sarah; Bermudez, Rey; Schensul, Jean

    2006-01-01

    Trends in ecstasy use in America during the past decade were reflected in mainstream, American rap-music lyrics between 1996 and 2003. Drawing on communication and cultural studies theory, this article provides a content analysis of 69 rap songs mentioning the club drug ecstasy. The songs are coded according to whether they contain positive, mixed…

  20. Ethnic Identity, Self-Esteem and Variability in Perceptions of Rap Music's Empowering and Risky Influences

    ERIC Educational Resources Information Center

    Travis, Raphael; Bowman, Scott W.

    2012-01-01

    Violence, risky sexual behaviors, and substance use are critical targets for improved health behavior. Prior research has linked levels of exposure to rap music with a range of undesirable health behaviors. Contemporary research has also found health-enhancing and other "positive" correlations with rap music exposure. The present study examined…

  1. Species-Specific Expression of Full-Length and Alternatively Spliced Variant Forms of CDK5RAP2

    PubMed Central

    Park, John S. Y.; Lee, Marie-Katrina; Kang, SungMyung; Jin, Yan; Fu, Songbin; Rosales, Jesusa L.; Lee, Ki-Young

    2015-01-01

    CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation. We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32. The equivalent of hCDK5RAP2-V1 has been reported in rat and mouse but the presence of full-length equivalent hCDK5RAP2 in rat and mouse has not been examined. Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively. However, mouse expresses only one form of CDK5RAP2 that is equivalent to the human and rat alternatively spliced variant forms. Knowledge of this expression of different forms of CDK5RAP2 in human, rat and mouse is essential in selecting the appropriate model for studies of CDK5RAP2 and primary microcephaly but our findings further indicate the evolutionary divergence of mouse from the human and rat species. PMID:26550838

  2. Species-Specific Expression of Full-Length and Alternatively Spliced Variant Forms of CDK5RAP2.

    PubMed

    Park, John S Y; Lee, Marie-Katrina; Kang, SungMyung; Jin, Yan; Fu, Songbin; Rosales, Jesusa L; Lee, Ki-Young

    2015-01-01

    CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation. We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32. The equivalent of hCDK5RAP2-V1 has been reported in rat and mouse but the presence of full-length equivalent hCDK5RAP2 in rat and mouse has not been examined. Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively. However, mouse expresses only one form of CDK5RAP2 that is equivalent to the human and rat alternatively spliced variant forms. Knowledge of this expression of different forms of CDK5RAP2 in human, rat and mouse is essential in selecting the appropriate model for studies of CDK5RAP2 and primary microcephaly but our findings further indicate the evolutionary divergence of mouse from the human and rat species. PMID:26550838

  3. 40 CFR 270.180 - For what reasons may the Director choose to revoke and reissue my final RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... choose to revoke and reissue my final RAP? 270.180 Section 270.180 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.180 For what reasons may the Director choose to revoke and reissue my final...

  4. 40 CFR 270.180 - For what reasons may the Director choose to revoke and reissue my final RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... choose to revoke and reissue my final RAP? 270.180 Section 270.180 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.180 For what reasons may the Director choose to revoke and reissue my final...

  5. 40 CFR 270.180 - For what reasons may the Director choose to revoke and reissue my final RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... choose to revoke and reissue my final RAP? 270.180 Section 270.180 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.180 For what reasons may the Director choose to revoke and reissue my final...

  6. 40 CFR 270.180 - For what reasons may the Director choose to revoke and reissue my final RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... choose to revoke and reissue my final RAP? 270.180 Section 270.180 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.180 For what reasons may the Director choose to revoke and reissue my final...

  7. 40 CFR 270.180 - For what reasons may the Director choose to revoke and reissue my final RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... choose to revoke and reissue my final RAP? 270.180 Section 270.180 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.180 For what reasons may the Director choose to revoke and reissue my final...

  8. Solution structure of the carboxyl-terminal domain of RAP74 and NMR characterization of the FCP1-binding sites of RAP74 and human TFIIB.

    PubMed

    Nguyen, Bao D; Chen, Hung-Ta; Kobor, Michael S; Greenblatt, Jack; Legault, Pascale; Omichinski, James G

    2003-02-18

    FCP1 (TFIIF-associated CTD phosphatase) is the only known phosphatase specific for the phosphorylated CTD of RNAP II. The phosphatase activity of FCP1 is strongly enhanced by the carboxyl-terminal domain of RAP74 (cterRAP74, residues 436-517), and this stimulatory effect of TFIIF can be blocked by TFIIB. It has been shown that cterRAP74 and the core domain of hTFIIB (TFIIBc, residues 112-316) directly interact with the carboxyl-terminal domain of hFCP1 (cterFCP, residues 879-961), and these interactions may be responsible for the regulatory activities of TFIIF and TFIIB on FCP1. We have determined the NMR solution structure of human cterRAP74, and we have used NMR methods to map the cterFCP-binding sites for both cterRAP74 and human TFIIB. We show that cterFCP binds to a groove of cterRAP74 between alpha-helices H2 and H3, without affecting the secondary structure of cterRAP74. We also show that cterFCP binds to a groove of TFIIBc between alpha-helices D1 and E1 in the first cyclin repeat. We find that the cterFCP-binding site of TFIIBc is very similar to the binding site for the HSV transcriptional activator protein VP16 on the first cyclin repeat of TFIIBc. The cterFCP-binding sites of both RAP74 and TFIIBc form shallow grooves on the protein surface, and they are both rich in hydrophobic and positively charged amino acid residues. These results provide new information about the recognition of acidic-rich activation domains involved in transcriptional regulation, and provide insights into how TFIIF and TFIIB regulate the FCP1 phosphatase activity in vivo. PMID:12578358

  9. 40 CFR 270.145 - What are the procedures for public comment on the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... comment on the draft RAP or notice of intent to deny? 270.145 Section 270.145 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.145 What are the procedures for public comment on the draft RAP or notice of intent to deny? (a) The Director must: (1)...

  10. 40 CFR 270.140 - What else must the Director prepare in addition to the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... addition to the draft RAP or notice of intent to deny? 270.140 Section 270.140 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.140 What else must the Director prepare in addition to the draft RAP or notice of intent to deny? Once the Director...

  11. 40 CFR 270.205 - What happens if I have applied correctly for a RAP renewal but have not received approval by the...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... correctly for a RAP renewal but have not received approval by the time my old RAP expires? 270.205 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.205 What happens if I have...

  12. 40 CFR 270.230 - May I perform remediation waste management activities under a RAP at a location removed from the...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... management activities under a RAP at a location removed from the area where the remediation wastes originated... Plans (RAPs) Obtaining A Rap for An Off-Site Location § 270.230 May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  13. 40 CFR 270.205 - What happens if I have applied correctly for a RAP renewal but have not received approval by the...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... correctly for a RAP renewal but have not received approval by the time my old RAP expires? 270.205 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.205 What happens if I have...

  14. 40 CFR 270.205 - What happens if I have applied correctly for a RAP renewal but have not received approval by the...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... correctly for a RAP renewal but have not received approval by the time my old RAP expires? 270.205 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.205 What happens if I have...

  15. 40 CFR 270.140 - What else must the Director prepare in addition to the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... addition to the draft RAP or notice of intent to deny? 270.140 Section 270.140 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.140 What else must the Director prepare in addition to the draft RAP or notice of intent to deny? Once the Director...

  16. 40 CFR 270.140 - What else must the Director prepare in addition to the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... addition to the draft RAP or notice of intent to deny? 270.140 Section 270.140 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.140 What else must the Director prepare in addition to the draft RAP or notice of intent to deny? Once the Director...

  17. 40 CFR 270.205 - What happens if I have applied correctly for a RAP renewal but have not received approval by the...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... correctly for a RAP renewal but have not received approval by the time my old RAP expires? 270.205 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.205 What happens if I have...

  18. 40 CFR 270.145 - What are the procedures for public comment on the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... comment on the draft RAP or notice of intent to deny? 270.145 Section 270.145 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.145 What are the procedures for public comment on the draft RAP or notice of intent to deny? (a) The Director must: (1)...

  19. 40 CFR 270.145 - What are the procedures for public comment on the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... comment on the draft RAP or notice of intent to deny? 270.145 Section 270.145 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.145 What are the procedures for public comment on the draft RAP or notice of intent to deny? (a) The Director must: (1)...

  20. 40 CFR 270.230 - May I perform remediation waste management activities under a RAP at a location removed from the...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... management activities under a RAP at a location removed from the area where the remediation wastes originated... Plans (RAPs) Obtaining A Rap for An Off-Site Location § 270.230 May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  1. 40 CFR 270.140 - What else must the Director prepare in addition to the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... addition to the draft RAP or notice of intent to deny? 270.140 Section 270.140 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.140 What else must the Director prepare in addition to the draft RAP or notice of intent to deny? Once the Director...

  2. 40 CFR 270.230 - May I perform remediation waste management activities under a RAP at a location removed from the...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... management activities under a RAP at a location removed from the area where the remediation wastes originated... Plans (RAPs) Obtaining A Rap for An Off-Site Location § 270.230 May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  3. 40 CFR 270.140 - What else must the Director prepare in addition to the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... addition to the draft RAP or notice of intent to deny? 270.140 Section 270.140 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.140 What else must the Director prepare in addition to the draft RAP or notice of intent to deny? Once the Director...

  4. 40 CFR 270.230 - May I perform remediation waste management activities under a RAP at a location removed from the...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... management activities under a RAP at a location removed from the area where the remediation wastes originated... Plans (RAPs) Obtaining A Rap for An Off-Site Location § 270.230 May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  5. 40 CFR 270.205 - What happens if I have applied correctly for a RAP renewal but have not received approval by the...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... correctly for a RAP renewal but have not received approval by the time my old RAP expires? 270.205 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) How May My Rap Be Modified, Revoked and Reissued, Or Terminated? § 270.205 What happens if I have...

  6. 40 CFR 270.145 - What are the procedures for public comment on the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... comment on the draft RAP or notice of intent to deny? 270.145 Section 270.145 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.145 What are the procedures for public comment on the draft RAP or notice of intent to deny? (a) The Director must: (1)...

  7. 40 CFR 270.145 - What are the procedures for public comment on the draft RAP or notice of intent to deny?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... comment on the draft RAP or notice of intent to deny? 270.145 Section 270.145 Protection of Environment... HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.145 What are the procedures for public comment on the draft RAP or notice of intent to deny? (a) The Director must: (1)...

  8. 40 CFR 270.230 - May I perform remediation waste management activities under a RAP at a location removed from the...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... management activities under a RAP at a location removed from the area where the remediation wastes originated... Plans (RAPs) Obtaining A Rap for An Off-Site Location § 270.230 May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  9. Characterization of low-temperature properties of plant-produced rap mixtures in the Northeast

    NASA Astrophysics Data System (ADS)

    Medeiros, Marcelo S., Junior

    The dissertation outlined herein results from a Federal Highway Administration sponsored project intended to investigate the impacts of high percentages of RAP material in the performance of pavements under cold climate conditions. It is comprised of two main sections that were incorporated into the body of this dissertation as Part I and Part II. In Part I a reduced testing framework for analysis of HMA mixes was proposed to replace the IDT creep compliance and strength testing by dynamic modulus and fatigue tests performed on an AMPT device. A continuum damage model that incorporates the nonlinear constitutive behavior of the HMA mixtures was also successfully implemented and validated. Mixtures with varying percentages of reclaimed material (RAP) ranging from 0 to 40% were used in this research effort in order to verify the applicability of the proposed methodology to RAP mixtures. Part II is concerned with evaluating the effects of various binder grades on the properties of plant-produced mixtures with various percentages of RAP. The effects of RAP on mechanical and rheological properties of mixtures and extracted binders were studied in order to identify some of the deficiencies in the current production methodologies. The results of this dissertation will help practitioners to identify optimal RAP usage from a material property perspective. It also establishes some guidelines and best practices for the use of higher RAP percentages in HMA.

  10. Numerical simulation of rip-raps with the distinct element method

    NASA Astrophysics Data System (ADS)

    Mittelbach, Livia

    2013-06-01

    and costal shores. They have to resist hydraulic loads such as ship and wind induced waves, tidal and ship induced currents, tidal varying water levels and storm surges. The numerical modelling of rip-rap revetments is undertaken by using the Distinct Element Method in three dimensions. With the DEM rip-rap stones can be modelled as autonomous objects with any degrees of freedom. Typical shapes of stones are formed by using clumped spherical particles. A method for the generation of the rip-rap stones based on geometrical and probabilistic parameters has been developed in order to generate stones with a realistic size and mass distribution. The DEM program is coupled with a computational fluid dynamics program to account for the influence of the hydraulic loads on the rip-rap stones. The acting forces can be simulated realistically for waves, currents and tidal varying water levels. Field measurements and model tests serve as validation for the numerical model. Physical model tests are carried out in a hydraulic flume with an instrumented rip-rap section for the calibration of the numerical stones material parameters. The behaviour of the particles depends on properties such as density, friction coefficient, normal and shear stiffness as well as the accuracy of the numerical representation of the rip-rap stones. Influences on the accuracy of the modelling of rip-raps with regard to the variation of these parameters are examined by comparing the results of the physical flume tests and numerical model.

  11. Sensitive detection and monitoring of senescence-associated secretory phenotype by SASP-RAP assay.

    PubMed

    Gu, Liubao; Kitamura, Masanori

    2012-01-01

    Senescence-associated secretory phenotype (SASP) is characterized by abundant secretion of various proteins in senescent cells and implicated in tumor progression and inflammatory responses. However, the profile of secreted proteins in SASP is different from cell type to cell type, and currently, universal markers for SASP have not been reported. In the present investigation, we show that SASP-responsive alkaline phosphatase (SASP-RAP) serves as a sensitive, general and convenient marker for SASP. Etoposide-treated cells exhibited a senescent phenotype characterized by senile morphology, positive staining for senescence-associated β-galactosidase, growth arrest and induction of p53 and p21(WAF1/CIP1). In SASP-RAP-transfected cells, exposure to etoposide increased secretion of SASP-RAP time-dependently. The kinetics of secretion was closely correlated with that of activation of the p21(WAF1/CIP1) promoter and the p16(INK4a) promoter. The enhanced secretion of SASP-RAP by senescence was also observed in cells treated with other senescence inducers such as trichostatin A, doxorubicin and 4-phenylbutylic acid. The induction of SASP-RAP by senescence was similarly observed in natural replicative senescence. To confirm selectivity of the SASP-RAP response, cells were treated with senescence-related and -unrelated stimuli (IL-1β, LPS, TNF-α and TGF-β), and induction of senescence markers and activity of SASP-RAP were evaluated in parallel. Unlike etoposide, senescence-unrelated stimuli did not induce p53 and p21(WAF1/CIP1), and it was correlated with lack of induction of SASP-RAP. In contrast, senescence-unrelated stimuli up-regulated conventional indicators for SASP, e.g., MMP-3, IL-6 and TIMP, without induction of senescence. SASP-RAP thus serves as a selective, convenient and general marker for detection and monitoring of SASP during cellular senescence.

  12. Early molecular and behavioral response to lipopolysaccharide in the WAG/Rij rat model of absence epilepsy and depressive-like behavior, involves interplay between AMPK, AKT/mTOR pathways and neuroinflammatory cytokine release.

    PubMed

    Russo, Emilio; Andreozzi, Francesco; Iuliano, Rodolfo; Dattilo, Vincenzo; Procopio, Teresa; Fiume, Giuseppe; Mimmi, Selena; Perrotti, Nicola; Citraro, Rita; Sesti, Giorgio; Constanti, Andrew; De Sarro, Giovambattista

    2014-11-01

    The mammalian target of rapamycin (mTOR) pathway has been recently indicated as a suitable drug target for the prevention of epileptogenesis. The mTOR pathway is known for its involvement in the control of the immune system. Since neuroinflammation is recognized as a major contributor to epileptogenesis, we wished to examine whether the neuroprotective effects of mTOR modulation could involve a suppression of the neuroinflammatory process in epileptic brain. We have investigated the early molecular mechanisms involved in the effects of intracerebral administration of the lipopolysaccharide (LPS) in the WAG/Rij rat model of absence epilepsy, in relation to seizure generation and depressive-like behavior; we also tested whether the effects of LPS could be modulated by treatment with rapamycin (RAP), a specific mTOR inhibitor. We determined, in specific rat brain areas, levels of p-mTOR/p-p70S6K and also p-AKT/p-AMPK as downstream or upstream indicators of mTOR activity and tested the effects of LPS and RAP co-administration. Changes in the brain levels of pro-inflammatory cytokines IL-1β and TNF-α and their relative mRNA expression levels were measured, and the involvement of nuclear factor-κB (NF-κB) was also examined in vitro. We confirmed that RAP inhibits the aggravation of absence seizures and depressive-like/sickness behavior induced by LPS in the WAG/Rij rats through the activation of mTOR and show that this effect is correlated with the ability of RAP to dampen and delay LPS increases in neuroinflammatory cytokines IL-1β and TNF-α, most likely through inhibition of the activation of NF-κB. Our results suggest that such a mechanism could contribute to the antiseizure, antiepileptogenic and behavioral effects of RAP and further highlight the potential therapeutic usefulness of mTOR inhibition in the management of human epilepsy and other neurological disorders. Furthermore, we show that LPS-dependent neuroinflammatory effects are also mediated by a

  13. EEG and MEG source localization using recursively applied (RAP) MUSIC

    SciTech Connect

    Mosher, J.C.; Leahy, R.M.

    1996-12-31

    The multiple signal characterization (MUSIC) algorithm locates multiple asynchronous dipolar sources from electroencephalography (EEG) and magnetoencephalography (MEG) data. A signal subspace is estimated from the data, then the algorithm scans a single dipole model through a three-dimensional head volume and computes projections onto this subspace. To locate the sources, the user must search the head volume for local peaks in the projection metric. Here we describe a novel extension of this approach which we refer to as RAP (Recursively APplied) MUSIC. This new procedure automatically extracts the locations of the sources through a recursive use of subspace projections, which uses the metric of principal correlations as a multidimensional form of correlation analysis between the model subspace and the data subspace. The dipolar orientations, a form of `diverse polarization,` are easily extracted using the associated principal vectors.

  14. Flight calibration assessment of HiRAP accelerometer data

    NASA Technical Reports Server (NTRS)

    Blanchard, Robert C.; Larman, Kevin T.; Moast, Christina D.

    1993-01-01

    A flight derived method of calibrating the High Resolution Accelerometer Package (HiRAP) flight data has been developed and is discussed for Shuttle Orbiter missions STS-35 and STS-40. These two mission data sets have been analyzed using ground calibration factors and flight derived calibration factors. This flight technique evolved early in the flight program when it was recognized that ground calibration factors are insufficient to determine absolute low-acceleration levels. The application of flight calibration factors to the data sets from these missions produced calibrated acceleration levels within an accuracy of less than +/- 1.5 microgravity of zero during a time in the flight when the acceleration level was known to be less than 1.0 microgravity. This analysis further confirms the theory that flight calibrations are required in order to obtain the absolute measurement of low-frequency, low-acceleration flight signals.

  15. Region 1: Radiological Assistance Program (RAP). Revision 2, Part 1

    SciTech Connect

    Hull, A.P.; Kuehner, A.V.

    1993-10-01

    The Department of Energy`s Radiological Assistance Program (RAP) is established under DOE Order 5530.3 to: (a) Establish and maintain response plans and resources to provide radiological assistance to other Federal agencies, State, local, and tribal governments, and private groups requesting such assistance. (b) Assist State, local, and tribal jurisdictions in preparing for radiological emergencies. (c) In the event of a real, or potential radiological accident, provide resources and monitoring and assessment assistance to other federal agencies, State, local, and tribal Governments. This plan is an integral part of a nationwide program of regionally based radiological assistance which has been established by DOE. The Brookhaven Area Office is the Regional Coordinating Office (RCO) for the Radiological Assistance Program in DOE Region 1, which consists of the New England States, New York, New Jersey, Pennsylvania, Delaware, Maryland and the District of Columbia.

  16. Crucial role of the Rap G protein signal in Notch activation and leukemogenicity of T-cell acute lymphoblastic leukemia.

    PubMed

    Doi, Keiko; Imai, Takahiko; Kressler, Christopher; Yagita, Hideo; Agata, Yasutoshi; Vooijs, Marc; Hamazaki, Yoko; Inoue, Joe; Minato, Nagahiro

    2015-01-23

    The Rap G protein signal regulates Notch activation in early thymic progenitor cells, and deregulated Rap activation (Rap(high)) results in the development of Notch-dependent T-cell acute lymphoblastic leukemia (T-ALL). We demonstrate that the Rap signal is required for the proliferation and leukemogenesis of established Notch-dependent T-ALL cell lines. Attenuation of the Rap signal by the expression of a dominant-negative Rap1A17 or Rap1GAP, Sipa1, in a T-ALL cell line resulted in the reduced Notch processing at site 2 due to impaired maturation of Adam10. Inhibition of the Rap1 prenylation with a geranylgeranyl transferase inhibitor abrogated its membrane-anchoring to Golgi-network and caused reduced proprotein convertase activity required for Adam10 maturation. Exogenous expression of a mature form of Adam10 overcame the Sipa1-induced inhibition of T-ALL cell proliferation. T-ALL cell lines expressed Notch ligands in a Notch-signal dependent manner, which contributed to the cell-autonomous Notch activation. Although the initial thymic blast cells barely expressed Notch ligands during the T-ALL development from Rap(high) hematopoietic progenitors in vivo, the ligands were clearly expressed in the T-ALL cells invading extrathymic vital organs. These results reveal a crucial role of the Rap signal in the Notch-dependent T-ALL development and the progression.

  17. The multifunctional transcription factor Rap1: a regulator of yeast physiology.

    PubMed

    Azad, Gajendra Kumar; Tomar, Raghuvir Singh

    2016-01-01

    Transcription is a fundamental process that is tightly regulated by transcription factors to maintain cellular homeostasis. Transcription factors have DNA-binding domains, some of which are sequence specific, and are found throughout the eukaryotic kingdom. Recent studies have revealed the molecular mechanisms by which transcription factors perform their functions. In the budding yeast Saccharomyces cerevisiae, Rap1 (ScRap1) can either activate or repress transcription. This bimodal transcriptional activity has led to the widespread study of the mode of action of ScRap1. This review summarizes current knowledge about yeast ScRap1, including its structure, mechanisms of transcription regulation, and biological functions, and the future directions in the field.

  18. The Ras/Rap GTPase activating protein RASA3: from gene structure to in vivo functions.

    PubMed

    Schurmans, Stéphane; Polizzi, Séléna; Scoumanne, Ariane; Sayyed, Sufyan; Molina-Ortiz, Patricia

    2015-01-01

    RASA3 (or GTPase Activating Protein III, R-Ras GTPase-activating protein, GAP1(IP4BP)) is a GTPase activating protein of the GAP1 subfamily which targets Ras and Rap1. RASA3 was originally purified from pig platelet membranes through its intrinsic ability to bind inositol 1,3,4,5-tetrakisphosphate (I(1,3,4,5)P4) with high affinity, hence its first name GAP1(IP4BP) (for GAP1 subfamily member which binds I(1,3,4,5)P4). RASA3 was thus the first I(1,3,4,5)P4 receptor identified and cloned. The in vitro and in vivo functions of RASA3 remained somewhat elusive for a long time. However, recently, using genetically-modified mice and cells derived from these mice, the function of RASA3 during megakaryopoiesis, megakaryocyte adhesion and migration as well as integrin signaling has been reported. The goal of this review is thus to summarize and comment recent and less recent data in the literature on RASA3, in particular on the in vivo function of this specific GAP1 subfamily member.

  19. Biological characterization of Drosophila Rapgap1, a GTPase activating protein for Rap1.

    PubMed

    Chen, F; Barkett, M; Ram, K T; Quintanilla, A; Hariharan, I K

    1997-11-11

    The activity of Ras family proteins is modulated in vivo by the function of GTPase activating proteins, which increase their intrinsic rate of GTP hydrolysis. We have isolated cDNAs encoding a GAP for the Drosophila Rap1 GTPase. Drosophila Rapgap1 encodes an 850-amino acid protein with a central region that displays substantial sequence similarity to human RapGAP. This domain, when expressed in Escherichia coli, potently stimulates Rap1 GTPase activity in vitro. Unlike Rap1, which is ubiquitously expressed, Rapgap1 expression is highly restricted. Rapgap1 is expressed at high levels in the developing photoreceptor cells and in the optic lobe. Rapgap1 mRNA is also localized in the pole plasm in an oskar-dependent manner. Although mutations that completely abolish Rapgap1 function display no obvious phenotypic abnormalities, overexpression of Rapgap1 induces a rough eye phenotype that is exacerbated by reducing Rap1 gene dosage. Thus, Rapgap1 can function as a negative regulator of Rap1-mediated signaling in vivo.

  20. Molecular Basis for Phosphorylation-dependent SUMO Recognition by the DNA Repair Protein RAP80.

    PubMed

    Anamika; Spyracopoulos, Leo

    2016-02-26

    Recognition and repair of double-stranded DNA breaks (DSB) involves the targeted recruitment of BRCA tumor suppressors to damage foci through binding of both ubiquitin (Ub) and the Ub-like modifier SUMO. RAP80 is a component of the BRCA1 A complex, and plays a key role in the recruitment process through the binding of Lys(63)-linked poly-Ub chains by tandem Ub interacting motifs (UIM). RAP80 also contains a SUMO interacting motif (SIM) just upstream of the tandem UIMs that has been shown to specifically bind the SUMO-2 isoform. The RAP80 tandem UIMs and SIM function collectively for optimal recruitment of BRCA1 to DSBs, although the molecular basis of this process is not well understood. Using NMR spectroscopy, we demonstrate that the RAP80 SIM binds SUMO-2, and that both specificity and affinity are enhanced through phosphorylation of the canonical CK2 site within the SIM. The affinity increase results from an enhancement of electrostatic interactions between the phosphoserines of RAP80 and the SIM recognition module within SUMO-2. The NMR structure of the SUMO-2·phospho-RAP80 complex reveals that the molecular basis for SUMO-2 specificity is due to isoform-specific sequence differences in electrostatic SIM recognition modules.

  1. Ability of Mn2+ to Permeate the Eye and Availability of Manganese-enhanced Magnetic Resonance Imaging for Visual Pathway Imaging via Topical Administration

    PubMed Central

    Chen, Yao; Shi, Chun-Yan; Li, Ying; Hu, Yun-Tao; Han, Hong-Bin; Sun, Xiao-Dong; Salvi, Satyajeet S; Ma, Zhi-Zhong

    2016-01-01

    Background: Manganese-enhanced magnetic resonance imaging (MEMRI) for visual pathway imaging via topical administration requires further research. This study investigated the permeability of the corneal epithelium and corneal toxicity after topical administration of Mn2+ to understand the applicability of MEMRI. Methods: Forty New Zealand rabbits were divided into 0.05 mol/L, 0.10 mol/L, and 0.20 mol/L groups as well as a control group (n = 10 in each group). Each group was further subdivided into epithelium-removed and epithelium-intact subgroups (n = 5 in each subgroup). Rabbits were given 8 drops of MnCl2 in 5 min intervals. The Mn2+ concentrations in the aqueous and vitreous humors were analyzed using inductively coupled plasma-mass spectrometry at different time points. MEMRI scanning was carried out to image the visual pathway after 24 h. The corneal toxicity of Mn2+ was evaluated with corneal imaging and pathology slices. Results: Between the aqueous and vitreous humors, there was a 10 h lag for the peak Mn2+ concentration times. The intraocular Mn2+ concentration increased with the concentration gradients of Mn2+ and was higher in the epithelium-removed subgroup than that in the epithelium-intact subgroup. The enhancement of the visual pathway was achieved in the 0.10 mol/L and 0.20 mol/L epithelium-removed subgroups. The corresponding peak concentrations of Mn2+ were 5087 ± 666 ng/ml, 22920 ± 1188 ng/ml in the aqueous humor and 884 ± 78 ng/ml, 2556 ± 492 ng/ml in the vitreous body, respectively. Corneal injury was evident in the epithelium-removed and 0.20 mol/L epithelium-intact subgroups. Conclusions: The corneal epithelium is a barrier to Mn2+, and the iris and lens septum might be another intraocular barrier to the permeation of Mn2+. An elevated Mn2+ concentration contributes to the increased permeation of Mn2+, higher MEMRI signal, and corneal toxicity. The enhancement of the visual pathway requires an effective Mn2+ concentration in the vitreous

  2. RAP1GA1: A candidate tumor suppressor locus in 1p36.1

    SciTech Connect

    Ranade, K.; Hussussian, C.J.; Higgins, P.

    1994-09-01

    The rap1/Krev-1 gene (RAP1A) encodes a p21-related protein that suppresses transformation by activated p21{sup ras}. The GTPase activating protein (GAP) gene for p21{sup rap1A} (RAP1GA1) has recently been assigned to chromosome 1p36.1-p35, a region of the genome that is frequently involved in deletions and rearrangements in several different tumors including breast, colon and hepatocellular carcinomas, melanoma, and neuroblastoma. GAP genes negatively regulate the activity of p21 proteins by catalyzing the conversion of the active GTP-bound forms to the inactive GDP-bound forms. The physiological function of p21{sup rap1A}-GAP makes it a strong candidate as a tumor suppressor gene that may have a role in the development of one or more of these malignancies. We have refined the localization of RAP1GA1 by linkage analysis with a highly informative (CA){sub n} repeat contained within the gene, and demonstrated that it is within the minimal deleted region for breast and colon carcinomas, and that it is excluded from the minimally deleted region in melanoma and neuroblastoma. Genetic mapping in the mouse demonstrated that Rap1ga1 is located {approximately}10 cM proximal to Pnd and therefore maps within the interval containing the modifier of Min gene (Mom-1) and the plasmocytoma susceptibility locus (Pcts). The human RAP1GA1 gene contains at least 27 exons. The coding region contains 22 exons, and there are at least five 5{prime}-UT exons that are assembled in a complex pattern of alternative splicing in different tissues. The localization of RAP1GA1 makes it a very strong candidate for a role as a modifier gene involved in the common secondary abnormalities involving 1p36 in several different carcinomas. The potential role of RAP1GA1 in these malignancies is currently being investigated by sequence analysis of breast and colon carcinomas with loss of heterozygosity in 1p36.

  3. Enhanced binding of RNAP II CTD phosphatase FCP1 to RAP74 following CK2 phosphorylation.

    PubMed

    Abbott, Karen L; Renfrow, Matthew B; Chalmers, Michael J; Nguyen, Bao D; Marshall, Alan G; Legault, Pascale; Omichinski, James G

    2005-03-01

    FCP1 (TFIIF-associated CTD phosphatase) is the first identified CTD-specific phosphatase required to recycle RNA polymerase II (RNAP II). FCP1 activity has been shown to be regulated by the general transcription factors TFIIF (RAP74) and TFIIB, protein kinase CK2 (CK2), and the HIV-1 transcriptional activator Tat. Phosphorylation of FCP1 by CK2 stimulates FCP1 phosphatase activity and enhances binding of RAP74 to FCP1. We have examined consensus CK2 phosphorylation sites (acidic residue n + 3 to serine or threonine residue) located immediately adjacent to both RAP74-binding sites of FCP1. We demonstrate that both of these consensus CK2 sites can be phosphorylated in vitro and that phosphorylation at either CK2 site results in enhanced binding of RAP74 to FCP1. The CK2 site adjacent to the RAP74-binding site in the central domain of FCP1 is phosphorylated at a single threonine site (T584). The CK2 site adjacent to the RAP74-binding site in the carboxyl-terminal domain can be phosphorylated at three successive serine residues (S942-S944), with phosphorylations at S942 and S944 both contributing to enhanced binding to RAP74. With the use of tandem Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR), we demonstrate that the phosphorylation of S942-S944 occurs in a semiordered fashion with the initial phosphorylation occurring at either S942 or S944 followed by a second phosphorylation to yield the S942/S944 diphosphorylated species. Using nuclear magnetic resonance (NMR) spectroscopy, we identify and map chemical shift changes onto the solution structure of the carboxyl-terminal domain of RAP74 (RAP74(436)(-)(517)) on complexation of RAP74(436)(-)(517) with phosphorylated FCP1 peptides. These results provide new functional and structural information on the role of phosphorylation in the recognition of acidic-rich activation domains involved in transcriptional regulation, and bring insights into how CK2 and TFIIF regulate FCP1 function. PMID:15723518

  4. RAP-PCR fingerprinting reveals time-dependent expression of development-related genes following differentiation process of Bacillus thuringiensis.

    PubMed

    Huang, Tianpei; Yu, Xiaomin; Gelbič, Ivan; Guan, Xiong

    2015-09-01

    Gene expression profiles are important data to reveal the functions of genes putatively involved in crucial biological processes. RNA arbitrarily primed polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription polymerase chain reaction (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB), which likely acted as virulence factors. RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase required for the development of Myxococcus xanthus. RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. RAP5 showed significant homology to a uridine kinase that mediates phosphorylation of uridine and azauridine. RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (also known as ketopantoate hydroxymethyltransferase or PanB) involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of B. thuringiensis and unravel novel pathogenic genes.

  5. Interaction of CDK5RAP2 with EB1 to track growing microtubule tips and to regulate microtubule dynamics.

    PubMed

    Fong, Ka-Wing; Hau, Shiu-Yeung; Kho, Yik-Shing; Jia, Yue; He, Lisheng; Qi, Robert Z

    2009-08-01

    Mutations in cdk5rap2 are linked to autosomal recessive primary microcephaly, and attention has been paid to its function at centrosomes. In this report, we demonstrate that CDK5RAP2 localizes to microtubules and concentrates at the distal tips in addition to centrosomal localization. CDK5RAP2 interacts directly with EB1, a prototypic member of microtubule plus-end tracking proteins, and contains the basic and Ser-rich motif responsible for EB1 binding. The EB1-binding motif is conserved in the CDK5RAP2 sequences of chimpanzee, bovine, and dog but not in those of rat and mouse, suggesting a function gained during the evolution of mammals. The mutation of the Ile/Leu-Pro dipeptide within the motif abolishes EB1 interaction and plus-end attachment. In agreement with the mutational analysis, suppression of EB1 expression inhibits microtubule tip-tracking of CDK5RAP2. We have also found that the CDK5RAP2-EB1 complex regulates microtubule dynamics and stability. CDK5RAP2 depletion by RNA interference impacts the dynamic behaviors of microtubules. The CDK5RAP2-EB1 complex induces microtubule bundling and acetylation when expressed in cell cultures and stimulates microtubule assembly and bundle formation in vitro. Collectively, these results show that CDK5RAP2 targets growing microtubule tips in association with EB1 to regulate microtubule dynamics. PMID:19553473

  6. Rap1 controls activation of the α(M)β(2) integrin in a talin-dependent manner.

    PubMed

    Lim, Jenson; Dupuy, Aurélien G; Critchley, David R; Caron, Emmanuelle

    2010-11-01

    The small GTPase Rap1 and the cytoskeletal protein talin regulate binding of C3bi-opsonised red blood cells (RBC) to integrin α(M)β(2) in phagocytic cells, although the mechanism has not been investigated. Using COS-7 cells transfected with α(M)β(2), we show that Rap1 acts on the β(2) and not the α(M) chain, and that residues 732-761 of the β(2) subunit are essential for Rap1-induced RBC binding. Activation of α(M)β(2) by Rap1 was dependent on W747 and F754 in the β(2) tails, which are required for talin head binding, suggesting a link between Rap1 and talin in this process. Using talin1 knock-out cells or siRNA-mediated talin1 knockdown in the THP-1 monocytic cell line, we show that Rap1 acts upstream of talin but surprisingly, RIAM knockdown had little effect on integrin-mediated RBC binding or cell spreading. Interestingly, Rap1 and talin influence each other's localisation at phagocytic cups, and co-immunoprecipitation experiments suggest that they interact together. These results show that Rap1-mediated activation of α(M)β(2) in macrophages shares both common and distinct features from Rap1 activation of α(IIb)β(3) expressed in CHO cells.

  7. An ultrasound-guided fascia iliaca catheter technique does not impair ambulatory ability within a clinical pathway for total hip arthroplasty

    PubMed Central

    Mudumbai, Seshadri C.; Kim, T. Edward; Howard, Steven K.; Giori, Nicholas J.; Woolson, Steven; Ganaway, Toni; Kou, Alex; King, Robert

    2016-01-01

    Background Both neuraxial and peripheral regional analgesic techniques offer postoperative analgesia for total hip arthroplasty (THA) patients. While no single technique is preferred, quadriceps muscle weakness from peripheral nerve blocks may impede rehabilitation. We designed this study to compare postoperative ambulation outcome in THA patients who were treated with a new ultrasound-guided fascia iliaca catheter (FIC) technique or intrathecal morphine (ITM). Methods We reviewed the electronic health records of a sequential series of primary unilateral THA patients who were part of a standardized clinical pathway; apart from differences in regional analgesic technique, all other aspects of the pathway were the same. Our primary outcome was total ambulation distance (meters) combined for postoperative days 1 and 2. Secondary outcomes included daily opioid consumption (morphine milligram equivalents) and analgesic-related side effects. We examined the association between the primary outcome and analgesic technique by performing crude and adjusted ordinary least-squares linear regression. A P value < 0.05 was considered statistically-significant. Results The study analyzed the records of 179 patients (fascia iliaca, n = 106; intrathecal, n = 73). The primary outcome (total ambulation distance) did not differ between the groups (P = 0.08). Body mass index (BMI) was the only factor (β = -1.7 [95% CI -0.5 to -2.9], P < 0.01) associated with ambulation distance. Opioid consumption did not differ, while increased pruritus was seen in the intrathecal group (P < 0.01). Conclusions BMI affects postoperative ambulation outcome after hip arthroplasty, whereas the type of regional analgesic technique used does not. An ultrasound-guided FIC technique offers similar analgesia with fewer side effects when compared with ITM. PMID:27482314

  8. Conserved motif of CDK5RAP2 mediates its localization to centrosomes and the Golgi complex.

    PubMed

    Wang, Zhe; Wu, Tao; Shi, Lin; Zhang, Lin; Zheng, Wei; Qu, Jianan Y; Niu, Ruifang; Qi, Robert Z

    2010-07-16

    As the primary microtubule-organizing centers, centrosomes require gamma-tubulin for microtubule nucleation and organization. Located in close vicinity to centrosomes, the Golgi complex is another microtubule-organizing organelle in interphase cells. CDK5RAP2 is a gamma-tubulin complex-binding protein and functions in gamma-tubulin attachment to centrosomes. In this study, we find that CDK5RAP2 localizes to the Golgi complex in an ATP- and centrosome-dependent manner and associates with Golgi membranes independently of microtubules. CDK5RAP2 contains a centrosome-targeting domain with its core region highly homologous to the Motif 2 (CM2) of centrosomin, a functionally related protein in Drosophila. This sequence, referred to as the CM2-like motif, is also conserved in related proteins in chicken and zebrafish. Therefore, CDK5RAP2 may undertake a conserved mechanism for centrosomal localization. Using a mutational approach, we demonstrate that the CM2-like motif plays a crucial role in the centrosomal and Golgi localization of CDK5RAP2. Furthermore, the CM2-like motif is essential for the association of the centrosome-targeting domain to pericentrin and AKAP450. The binding with pericentrin is required for the centrosomal and Golgi localization of CDK5RAP2, whereas the binding with AKAP450 is required for the Golgi localization. Although the CM2-like motif possesses the activity of Ca(2+)-independent calmodulin binding, binding of calmodulin to this sequence is dispensable for centrosomal and Golgi association. Altogether, CDK5RAP2 may represent a novel mechanism for centrosomal and Golgi localization. PMID:20466722

  9. RAP-011, an activin receptor ligand trap, increases hemoglobin concentration in hepcidin transgenic mice.

    PubMed

    Langdon, Jacqueline M; Barkataki, Sangjucta; Berger, Alan E; Cheadle, Chris; Xue, Qian-Li; Sung, Victoria; Roy, Cindy N

    2015-01-01

    Over expression of hepcidin antimicrobial peptide is a common feature of iron-restricted anemia in humans. We investigated the erythroid response to either erythropoietin or RAP-011, a "murinized" ortholog of sotatercept, in C57BL/6 mice and in hepcidin antimicrobial peptide 1 over expressing mice. Sotatercept, a soluble, activin receptor type IIA ligand trap, is currently being evaluated for the treatment of anemias associated with chronic renal disease, myelodysplastic syndrome, β-thalassemia, and Diamond Blackfan anemia and acts by inhibiting signaling downstream of activin and other Transforming Growth Factor-β superfamily members. We found that erythropoietin and RAP-011 increased hemoglobin concentration in C57BL/6 mice and in hepcidin antimicrobial peptide 1 over expressing mice. While erythropoietin treatment depleted splenic iron stores in C57BL/6 mice, RAP-011 treatment did not deplete splenic iron stores in mice of either genotype. Bone marrow erythroid progenitors from erythropoietin-treated mice exhibited iron-restricted erythropoiesis, as indicated by increased median fluorescence intensity of transferrin receptor immunostaining by flow cytometry. In contrast, RAP-011-treated mice did not exhibit the same degree of iron-restricted erythropoiesis. In conclusion, we have demonstrated that RAP-011 can improve hemoglobin concentration in hepcidin antimicrobial peptide 1 transgenic mice. Our data support the hypothesis that RAP-011 has unique biologic effects which prevent or circumvent depletion of mouse splenic iron stores. RAP-011 may, therefore, be an appropriate therapeutic for trials in human anemias characterized by increased expression of hepcidin antimicrobial peptide and iron-restricted erythropoiesis.

  10. Stearoyl lysophosphatidylcholine enhances the phagocytic ability of macrophages through the AMP-activated protein kinase/p38 mitogen activated protein kinase pathway.

    PubMed

    Quan, Hui; Hur, Young-Hoe; Xin, Chun; Kim, Joung-Min; Choi, Jeong-Il; Kim, Man-Young; Bae, Hong-Beom

    2016-10-01

    A previous study showed that stearoyl lysophosphatidylcholine (sLPC) suppressed extracellular high mobility group box 1 translocation in macrophages stimulated with lipopolysaccharide through AMP-activated protein kinase (AMPK) activation. In the present study, we investigated whether sLPC-induced AMPK activation could enhance macrophages phagocytosis of bacteria. We found that sLPC increased phosphorylation of AMPK and acetyl-CoA carboxylase, a downstream target of AMPK, in a time- and dose-dependent manner in macrophages. Furthermore, sLPC increased the uptake of FITC-conjugated Escherichia coli by macrophages in a dose-dependent manner, and treatment with an AMPK inhibitor (compound C) or siRNA to AMPKα1 reversed this uptake. sLPC increased the phosphorylation of p38 mitogen-activated protein kinase (MAPK), but inhibition of AMPK activity with compound C or siRNA to AMPKα1 prevented the sLPC-induced increase in p38 MAPK phosphorylation. SB203580, a p38 MAPK inhibitor, decreased sLPC-induced phagocytosis. In vivo, systemic administration of sLPC to mice led to increased AMPK and p38 MAPK activity in the lung and to increased phagocytosis of fluorescent E. coli in bronchoalveolar lavage cells. These results suggest that sLPC increases macrophages phagocytosis through activation of the AMPK/p38 MAPK pathway. Therefore, sLPC is a candidate pharmacological agent for the treatment of bacterial infections in clinically relevant conditions. PMID:27517519

  11. CDK5RAP2 functions in centrosome to spindle pole attachment and DNA damage response.

    PubMed

    Barr, Alexis R; Kilmartin, John V; Gergely, Fanni

    2010-04-01

    The centrosomal protein, CDK5RAP2, is mutated in primary microcephaly, a neurodevelopmental disorder characterized by reduced brain size. The Drosophila melanogaster homologue of CDK5RAP2, centrosomin (Cnn), maintains the pericentriolar matrix (PCM) around centrioles during mitosis. In this study, we demonstrate a similar role for CDK5RAP2 in vertebrate cells. By disrupting two evolutionarily conserved domains of CDK5RAP2, CNN1 and CNN2, in the avian B cell line DT40, we find that both domains are essential for linking centrosomes to mitotic spindle poles. Although structurally intact, centrosomes lacking the CNN1 domain fail to recruit specific PCM components that mediate attachment to spindle poles. Furthermore, we show that the CNN1 domain enforces cohesion between parental centrioles during interphase and promotes efficient DNA damage-induced G2 cell cycle arrest. Because mitotic spindle positioning, asymmetric centrosome inheritance, and DNA damage signaling have all been implicated in cell fate determination during neurogenesis, our findings provide novel insight into how impaired CDK5RAP2 function could cause premature depletion of neural stem cells and thereby microcephaly. PMID:20368616

  12. Cdk5rap2 regulates centrosome function and chromosome segregation in neuronal progenitors.

    PubMed

    Lizarraga, Sofia B; Margossian, Steven P; Harris, Marian H; Campagna, Dean R; Han, An-Ping; Blevins, Sherika; Mudbhary, Raksha; Barker, Jane E; Walsh, Christopher A; Fleming, Mark D

    2010-06-01

    Microcephaly affects approximately 1% of the population and is associated with mental retardation, motor defects and, in some cases, seizures. We analyzed the mechanisms underlying brain size determination in a mouse model of human microcephaly. The Hertwig's anemia (an) mutant shows peripheral blood cytopenias, spontaneous aneuploidy and a predisposition to hematopoietic tumors. We found that the an mutation is a genomic inversion of exon 4 of Cdk5rap2, resulting in an in-frame deletion of exon 4 from the mRNA. The finding that CDK5RAP2 human mutations cause microcephaly prompted further analysis of Cdk5rap2(an/an) mice and we demonstrated that these mice exhibit microcephaly comparable to that of the human disease, resulting from striking neurogenic defects that include proliferative and survival defects in neuronal progenitors. Cdk5rap2(an/an) neuronal precursors exit the cell cycle prematurely and many undergo apoptosis. These defects are associated with impaired mitotic progression coupled with abnormal mitotic spindle pole number and mitotic orientation. Our findings suggest that the reduction in brain size observed in humans with mutations in CDK5RAP2 is associated with impaired centrosomal function and with changes in mitotic spindle orientation during progenitor proliferation. PMID:20460369

  13. CDK5RAP2 regulates centriole engagement and cohesion in mice.

    PubMed

    Barrera, Jose A; Kao, Ling-Rong; Hammer, Robert E; Seemann, Joachim; Fuchs, Jannon L; Megraw, Timothy L

    2010-06-15

    Centriole duplication occurs once per cell cycle, ensuring that each cell contains two centrosomes, each containing a mother-daughter pair of tightly engaged centrioles at mitotic entry. Loss of the tight engagement between mother and daughter centrioles appears to license the next round of centriole duplication. However, the molecular mechanisms regulating this process remain largely unknown. Mutations in CDK5RAP2, which encodes a centrosomal protein, cause autosomal recessive primary microcephaly in humans. Here we show that CDK5RAP2 loss of function in mice causes centriole amplification with a preponderance of single, unpaired centrioles and increased numbers of daughter-daughter centriole pairs. These results indicate that CDK5RAP2 is required to maintain centriole engagement and cohesion, thereby restricting centriole replication. Early in mitosis, amplified centrosomes assemble multipolar spindles in CDK5RAP2 mutant cells. Moreover, both mother and daughter centrioles are amplified and the excess mother centrioles template multiple primary cilia in CDK5RAP2 mutant cells. PMID:20627074

  14. Fission yeast Cactin restricts telomere transcription and elongation by controlling Rap1 levels.

    PubMed

    Lorenzi, Luca E; Bah, Amadou; Wischnewski, Harry; Shchepachev, Vadim; Soneson, Charlotte; Santagostino, Marco; Azzalin, Claus M

    2015-01-01

    The telomeric transcriptome comprises multiple long non-coding RNAs generated by transcription of linear chromosome ends. In a screening performed in Schizosaccharomyces pombe, we identified factors modulating the cellular levels of the telomeric transcriptome. Among these factors, Cay1 is the fission yeast member of the conserved family of Cactins, uncharacterized proteins crucial for cell growth and survival. In cay1∆ mutants, the cellular levels of the telomeric factor Rap1 are drastically diminished due to defects in rap1+ pre-mRNA splicing and Rap1 protein stability. cay1∆ cells accumulate histone H3 acetylated at lysine 9 at telomeres, which become transcriptionally desilenced, are over-elongated by telomerase and cause chromosomal aberrations in the cold. Overexpressing Rap1 in cay1+ deleted cells significantly reverts all telomeric defects. Additionally, cay1∆ mutants accumulate unprocessed Tf2 retrotransposon RNA through Rap1-independent mechanisms. Thus, Cay1 plays crucial roles in cells by ultimately harmonizing expression of transcripts originating from seemingly unrelated genomic loci.

  15. Fission yeast Cactin restricts telomere transcription and elongation by controlling Rap1 levels

    PubMed Central

    Lorenzi, Luca E; Bah, Amadou; Wischnewski, Harry; Shchepachev, Vadim; Soneson, Charlotte; Santagostino, Marco; Azzalin, Claus M

    2015-01-01

    The telomeric transcriptome comprises multiple long non-coding RNAs generated by transcription of linear chromosome ends. In a screening performed in Schizosaccharomyces pombe, we identified factors modulating the cellular levels of the telomeric transcriptome. Among these factors, Cay1 is the fission yeast member of the conserved family of Cactins, uncharacterized proteins crucial for cell growth and survival. In cay1Δ mutants, the cellular levels of the telomeric factor Rap1 are drastically diminished due to defects in rap1+ pre-mRNA splicing and Rap1 protein stability. cay1Δ cells accumulate histone H3 acetylated at lysine 9 at telomeres, which become transcriptionally desilenced, are over-elongated by telomerase and cause chromosomal aberrations in the cold. Overexpressing Rap1 in cay1+ deleted cells significantly reverts all telomeric defects. Additionally, cay1Δ mutants accumulate unprocessed Tf2 retrotransposon RNA through Rap1-independent mechanisms. Thus, Cay1 plays crucial roles in cells by ultimately harmonizing expression of transcripts originating from seemingly unrelated genomic loci. PMID:25398909

  16. Changing images of violence in Rap music lyrics: 1979-1997.

    PubMed

    Herd, Denise

    2009-12-01

    Rap music has been at the center of concern about the potential harmful effects of violent media on youth social behavior. This article explores the role of changing images of violence in rap music lyrics from the 1970s to the 1990s. The results indicate that there has been a dramatic and sustained increase in the level of violence in rap music. The percentage of songs mentioning violence increased from 27 per cent during 1979-1984 to 60 per cent during 1994-1997. In addition, portrayals of violence in later songs are viewed in a more positive light as shown by their increased association with glamor, wealth, masculinity, and personal prowess. Additional analyses revealed that genre, specifically gangster rap, is the most powerful predictor of the increased number of violent references in songs. The discussion suggests that violence in rap music has increased in response to the complex interplay of changing social conditions such as the elevated levels of youth violence in the 1980s and changing commercial practices within the music industry.

  17. PLK1-dependent activation of LRRK1 regulates spindle orientation by phosphorylating CDK5RAP2.

    PubMed

    Hanafusa, Hiroshi; Kedashiro, Shin; Tezuka, Motohiro; Funatsu, Motoki; Usami, Satoshi; Toyoshima, Fumiko; Matsumoto, Kunihiro

    2015-08-01

    Correct formation of the cell division axis requires the initial precise orientation of the mitotic spindle. Proper spindle orientation depends on centrosome maturation, and Polo-like kinase 1 (PLK1) is known to play a crucial role in this process. However, the molecular mechanisms that function downstream of PLK1 are not well understood. Here we show that LRRK1 is a PLK1 substrate that is phosphorylated on Ser 1790. PLK1 phosphorylation is required for CDK1-mediated activation of LRRK1 at the centrosomes, and this in turn regulates mitotic spindle orientation by nucleating the growth of astral microtubules from the centrosomes. Interestingly, LRRK1 in turn phosphorylates CDK5RAP2(Cep215), a human homologue of Drosophila Centrosomin (Cnn), in its γ-tubulin-binding motif, thus promoting the interaction of CDK5RAP2 with γ-tubulin. LRRK1 phosphorylation of CDK5RAP2 Ser 140 is necessary for CDK5RAP2-dependent microtubule nucleation. Thus, our findings provide evidence that LRRK1 regulates mitotic spindle orientation downstream of PLK1 through CDK5RAP2-dependent centrosome maturation. PMID:26192437

  18. JAK tyrosine kinases promote hierarchical activation of Rho and Rap modules of integrin activation.

    PubMed

    Montresor, Alessio; Bolomini-Vittori, Matteo; Toffali, Lara; Rossi, Barbara; Constantin, Gabriela; Laudanna, Carlo

    2013-12-23

    Lymphocyte recruitment is regulated by signaling modules based on the activity of Rho and Rap small guanosine triphosphatases that control integrin activation by chemokines. We show that Janus kinase (JAK) protein tyrosine kinases control chemokine-induced LFA-1- and VLA-4-mediated adhesion as well as human T lymphocyte homing to secondary lymphoid organs. JAK2 and JAK3 isoforms, but not JAK1, mediate CXCL12-induced LFA-1 triggering to a high affinity state. Signal transduction analysis showed that chemokine-induced activation of the Rho module of LFA-1 affinity triggering is dependent on JAK activity, with VAV1 mediating Rho activation by JAKs in a Gαi-independent manner. Furthermore, activation of Rap1A by chemokines is also dependent on JAK2 and JAK3 activity. Importantly, activation of Rap1A by JAKs is mediated by RhoA and PLD1, thus establishing Rap1A as a downstream effector of the Rho module. Thus, JAK tyrosine kinases control integrin activation and dependent lymphocyte trafficking by bridging chemokine receptors to the concurrent and hierarchical activation of the Rho and Rap modules of integrin activation.

  19. APP independent and dependent effects on neurite outgrowth are modulated by the receptor associated protein (RAP).

    PubMed

    Billnitzer, Andrew J; Barskaya, Irina; Yin, Cailing; Perez, Ruth G

    2013-01-01

    Amyloid precursor protein (APP) and its secreted form, sAPP, contribute to the development of neurons in hippocampus, a brain region critical for learning and memory. Full-length APP binds the low-density lipoprotein receptor-related protein (LRP), which stimulates APP endocytosis. LRP also contributes to neurite growth. Furthermore, the receptor associated protein (RAP) binds LRP in a manner that blocks APP-LRP interactions. To elucidate APP contributions to neurite growth for full-length APP and sAPP, we cultured wild type (WT) and APP knockout (KO) neurons in sAPPα and/or RAP and measured neurite outgrowth at 1 day in vitro. Our data reveal that WT neurons had less axonal outgrowth including less axon branching. RAP treatment potentiated the inhibitory effects of APP. KO neurons had significantly more outgrowth and branching, especially in response to RAP, effects which were also associated with ERK2 activation. Our results affirm a major inhibitory role by full-length APP on all aspects of axonal and dendritic outgrowth, and show that RAP-LRP binding stimulated axon growth independently of APP. These findings support a major role for APP as an inhibitor of neurite growth and reveal novel signaling functions for LRP that may be disrupted by Alzheimer's pathology or therapies aimed at APP processing.

  20. Changing images of violence in Rap music lyrics: 1979-1997.

    PubMed

    Herd, Denise

    2009-12-01

    Rap music has been at the center of concern about the potential harmful effects of violent media on youth social behavior. This article explores the role of changing images of violence in rap music lyrics from the 1970s to the 1990s. The results indicate that there has been a dramatic and sustained increase in the level of violence in rap music. The percentage of songs mentioning violence increased from 27 per cent during 1979-1984 to 60 per cent during 1994-1997. In addition, portrayals of violence in later songs are viewed in a more positive light as shown by their increased association with glamor, wealth, masculinity, and personal prowess. Additional analyses revealed that genre, specifically gangster rap, is the most powerful predictor of the increased number of violent references in songs. The discussion suggests that violence in rap music has increased in response to the complex interplay of changing social conditions such as the elevated levels of youth violence in the 1980s and changing commercial practices within the music industry. PMID:20029428

  1. Rap1 integrates tissue polarity, lumen formation, and tumorigenicpotential in human breast epithelial cells

    SciTech Connect

    Itoh, Masahiko; Nelson, Celeste M.; Myers, Connie A.; Bissell,Mina J.

    2006-09-29

    Maintenance of apico-basal polarity in normal breast epithelial acini requires a balance between cell proliferation, cell death, and proper cell-cell and cell-extracellular matrix signaling. Aberrations in any of these processes can disrupt tissue architecture and initiate tumor formation. Here we show that the small GTPase Rap1 is a crucial element in organizing acinar structure and inducing lumen formation. Rap1 activity in malignant HMT-3522 T4-2 cells is appreciably higher than in S1 cells, their non-malignant counterparts. Expression of dominant-negative Rap1 resulted in phenotypic reversion of T4-2 cells, led to formation of acinar structures with correct apico-basal polarity, and dramatically reduced tumor incidence despite the persistence of genomic abnormalities. The resulting acini contained prominent central lumina not observed when other reverting agents were used. Conversely, expression of dominant-active Rap1 in T4-2 cells inhibited phenotypic reversion and led to increased invasiveness and tumorigenicity. Thus, Rap1 acts as a central regulator of breast architecture, with normal levels of activation instructing apical polarity during acinar morphogenesis, and increased activation inducing tumor formation and progression to malignancy.

  2. Hip Hop Therapy: An Exploratory Study of a Rap Music Intervention with At-Risk and Delinquent Youth.

    ERIC Educational Resources Information Center

    Tyson, Edgar H.

    2002-01-01

    Presents an exploratory study of the therapeutic potential of "Hip-Hop" therapy, an "innovative synergy of rap music, bibliotherapy, and music therapy." Finds that the quantitative and qualitative results partially supported the hypothesis that under a specific set of conditions rap music would improve the therapeutic experience and outcomes for…

  3. Feeling the Beat: The Meaning of Rap Music for Ethnically Diverse Midwestern College Students--A Phenomenological Study

    ERIC Educational Resources Information Center

    Iwamoto, Derek K.; Creswell, John; Caldwell, Leon

    2007-01-01

    Despite its national and international appeal, rap is considered one of the most controversial of music genres. Given the political charge it generates, rap music has spawned research across the social and health sciences. The majority of the research has investigated its impact on African Americans. Further, the research has tended to focus on…

  4. An Educational Exploration of Homophobia and Sexism in Rap and Hip Hop: Homo-Thugs and Divas in da House

    ERIC Educational Resources Information Center

    Chiu, Nicholas

    2005-01-01

    In this paper, the author explores the connections between hip hop and rap, sexism and homophobia, and children and teens. He describes the implications or potential consequences of sexism and homophobia within the music and media culture of hip hop and rap (with the focus on how it affects young viewers and fans in terms of gender [identity]…

  5. CDK5RAP2 is a pericentriolar protein that functions in centrosomal attachment of the gamma-tubulin ring complex.

    PubMed

    Fong, Ka-Wing; Choi, Yuk-Kwan; Rattner, Jerome B; Qi, Robert Z

    2008-01-01

    Microtubule nucleation and organization by the centrosome require gamma-tubulin, a protein that exists in a macromolecular complex called the gamma-tubulin ring complex (gammaTuRC). We report characterization of CDK5RAP2, a novel centrosomal protein whose mutations have been linked to autosomal recessive primary microcephaly. In somatic cells, CDK5RAP2 localizes throughout the pericentriolar material in all stages of the cell cycle. When overexpressed, CDK5RAP2 assembled a subset of centrosomal proteins including gamma-tubulin onto the centrosomes or under the microtubule-disrupting conditions into microtubule-nucleating clusters in the cytoplasm. CDK5RAP2 associates with the gammaTuRC via a short conserved sequence present in several related proteins found in a range of organisms from fungi to mammals. The binding of CDK5RAP2 is required for gammaTuRC attachment to the centrosome but not for gammaTuRC assembly. Perturbing CDK5RAP2 function delocalized gamma-tubulin from the centrosomes and inhibited centrosomal microtubule nucleation, thus leading to disorganization of interphase microtubule arrays and formation of anastral mitotic spindles. Together, CDK5RAP2 is a pericentriolar structural component that functions in gammaTuRC attachment and therefore in the microtubule organizing function of the centrosome. Our findings suggest that centrosome malfunction due to the CDK5RAP2 mutations may underlie autosomal recessive primary microcephaly. PMID:17959831

  6. Dual functions of Rap1 are crucial for T-cell homeostasis and prevention of spontaneous colitis

    PubMed Central

    Ishihara, Sayaka; Nishikimi, Akihiko; Umemoto, Eiji; Miyasaka, Masayuki; Saegusa, Makoto; Katagiri, Koko

    2015-01-01

    Rap1-GTP activates leukocyte function-associated antigen-1 (LFA-1) to induce arrest on the high endothelial venule (HEV). Here we show that Rap1-GDP restrains rolling behaviours of T cells on the peripheral lymph node addressin (PNAd), P-selectin and mucosal addressin cell adhesion molecule-1 (MadCAM-1) by inhibiting tether formation. Consequently, Rap1 deficiency impairs homing of naive T cells to peripheral lymph nodes, but accelerates homing of TH17 and TH1 cells to the colon, resulting in spontaneous colitis with tumours. Rap1-GDP associates with and activates lymphocyte-oriented kinase, which phosphorylates ERM (ezrin, radixin and moesin) in resting T cells. Phosphomimetic ezrin reduces the rolling of Rap1-deficient cells, and thereby decreases their homing into the colon. On the other hand, chemokines activate Rap1 at the plasma membrane within seconds, and Rap1-GTP binds to filamins, which diminishes its association with the β2 chain of LFA-1 and results in LFA-1 activation. This Rap1-dependent regulation of T-cell circulation prevents the onset of colitis. PMID:26634692

  7. Field performance of maintenance treatments constructed with reclaimed asphalt pavement (RAP). Final research report, September 1992-August 1994

    SciTech Connect

    Estakhri, C.K.

    1994-11-01

    In the study, RAP was blended with recycling emulsions and conventional maintenance mixtures in attempts to improve its field performance as a maintenance mixture. RAP was also mixed with stabilizers and used as a base material in maintenance projects. Several field experiments were constructed throughout the state, and the report documents their performance.

  8. A novel neutralization sensitive and subdominant RAP-1-related antigen (RRA) is expressed by babesia bovis merozoites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objective: The Babesia bovis genome encodes a rap-1 related gene denominated RAP-1 related antigen (RRA). In this study, we analyzed the pattern of expression, immunogenicity and functional relevance of RRA. Methods: Phylogenetic analysis was performed using the program Phylip. Expression of rra wa...

  9. The Immediate Effects of Homicidal, Suicidal, and Nonviolent Heavy Metal and Rap Songs on the Moods of College Students.

    ERIC Educational Resources Information Center

    Ballard, Mary E.; Coates, Steven

    1995-01-01

    Examined the impact of homicidal, suicidal, and nonviolent heavy metal and rap songs on the moods of male college undergraduates. Students (n=164) completed mood inventories after listening to 1 of 6 songs. Results show no effects of these songs on suicidal ideation, anxiety, or self-esteem. Rap songs elicited greater angry responses than heavy…

  10. RAP1 Is Essential for Silencing Telomeric Variant Surface Glycoprotein Genes in Trypanosoma brucei

    PubMed Central

    Yang, Xiaofeng; Figueiredo, Luisa M.; Espinal, Amin; Okubo, Eiji; Li, Bibo

    2009-01-01

    SUMMARY Trypanosoma brucei expresses Variant Surface Glycoprotein (VSG) genes in a strictly monoallelic fashion in its mammalian hosts, but it is unclear how this important virulence mechanism is enforced. Telomere position effect (TPE), an epigenetic phenomenon, has been proposed to play a critical role in VSG regulation, yet no telomeric protein has been identified whose disruption led to VSG derepression. We now identify tbRAP1 as an intrinsic component of the T. brucei telomere complex and a major regulator for silencing VSG expression sites (ESs). Knockdown of tbRAP1 led to derepression of all VSGs in silent ESs, but not VSGs located elsewhere, and resulted in stronger derepression of genes located within 10 kb from telomeres than genes located further upstream. This graduated silencing pattern suggests that telomere integrity plays a key role in tbRAP1-dependent silencing and VSG regulation. PMID:19345190

  11. Changes in the prevalence of alcohol in rap music lyrics 1979-2009.

    PubMed

    Herd, Denise

    2014-02-01

    This study examines the prevalence and context of alcohol references in rap music lyrics from 1979 through 2009. Four hundred nine top-ranked rap music songs released were sampled from Billboard magazine rating charts. Songs were analyzed using systematic content analysis and were coded for alcohol beverage types and brand names, drinking behaviors, drinking contexts, attitudes towards alcohol, and consequences of drinking. Trends were analyzed using regression analyses. The results of the study reveal significant increases in the presence of alcohol in rap songs; a decline in negative attitudes towards alcohol; decreases in consequences attributed to alcohol; increases in the association of alcohol with glamour and wealth, drugs, and nightclubs; and increases in references to liquor and champagne.

  12. Ser756 of β2 integrin controls Rap1 activity during inside-out activation of αMβ2.

    PubMed

    Lim, Jenson; Hotchin, Neil A; Caron, Emmanuelle

    2011-08-01

    During αMβ2-mediated phagocytosis, the small GTPase Rap1 activates the β2 integrin by binding to a region between residues 732 and 761. Using COS-7 cells transfected with αMβ2, we show that αMβ2 activation by the phorbol ester PMA involves Ser(756) of β2. This residue is critical for the local positioning of talin and biochemically interacts with Rap1. Using the CaM (calmodulin) antagonist W7, we found Rap1 recruitment and the inside-out activation of αMβ2 to be affected. We also report a role for CaMKII (calcium/CaM-dependent kinase II) in the activation of Rap1 during integrin activation. These results demonstrate a distinct physiological role for Ser(756) of β2 integrin, in conjunction with the actions of talin and Rap1, during αMβ2 activation in macrophages.

  13. Telomere protein RAP1 levels are affected by cellular aging and oxidative stress

    PubMed Central

    Swanson, Mark J.; Baribault, Michelle E.; Israel, Joanna N.; Bae, Nancy S.

    2016-01-01

    Telomeres are important for maintaining the integrity of the genome through the action of the shelterin complex. Previous studies indicted that the length of the telomere did not have an effect on the amount of the shelterin subunits; however, those experiments were performed using immortalized cells with stable telomere lengths. The interest of the present study was to observe how decreasing telomere lengths over successive generations would affect the shelterin subunits. As neonatal human dermal fibroblasts aged and their telomeres became shorter, the levels of the telomere-binding protein telomeric repeat factor 2 (TRF2) decreased significantly. By contrast, the levels of one of its binding partners, repressor/activator protein 1 (RAP1), decreased to a lesser extent than would be expected from the decrease in TRF2. Other subunits, TERF1-interacting nuclear factor 2 and protection of telomeres protein 1, remained stable. The decrease in RAP1 in the older cells occurred in the nuclear and cytoplasmic fractions. Hydrogen peroxide (H2O2) stress was used as an artificial means of aging in the cells, and this resulted in RAP1 levels decreasing, but the effect was only observed in the nuclear portion. Similar results were obtained using U251 glioblastoma cells treated with H2O2 or grown in serum-depleted medium. The present findings indicate that TRF2 and RAP1 levels decrease as fibroblasts naturally age. RAP1 remains more stable compared to TRF2. RAP1 also responds to oxidative stress, but the response is different to that observed in aging. PMID:27446538

  14. Structural and Functional Studies of the Rap1 C-Terminus Reveal Novel Separation-of-Function Mutants

    SciTech Connect

    Feeser, Elizabeth A.; Wolberger, Cynthia

    2010-02-19

    The yeast Rap1 protein plays an important role in transcriptional silencing and in telomere length homeostasis. Rap1 mediates silencing at the HM loci and at telomeres by recruiting the Sir3 and Sir4 proteins to chromatin via a Rap1 C-terminal domain, which also recruits the telomere length regulators, Rif1 and Rif2. We report the 1.85 {angstrom} resolution crystal structure of the Rap1 C-terminus, which adopts an all-helical fold with no structural homologues. The structure was used to engineer surface mutations in Rap1, and the effects of these mutations on silencing and telomere length regulation were assayed in vivo. Our surprising finding was that there is no overlap between mutations affecting mating-type and telomeric silencing, suggesting that Rap1 plays distinct roles in silencing at the silent mating-type loci and telomeres. We also found novel Rap1 phenotypes and new separation-of-function mutants, which provide new tools for studying Rap1 function. Yeast two-hybrid studies were used to determine how specific mutations affect recruitment of Sir3, Rif1, and Rif2. A comparison of the yeast two-hybrid and functional data reveals patterns of protein interactions that correlate with each Rap1 phenotype. We find that Sir3 interactions are important for telomeric silencing, but not mating type silencing, and that Rif1 and Rif2 interactions are important in different subsets of telomeric length mutants. Our results show that the role of Rap1 in silencing differs between the HM loci and the telomeres and offer insight into the interplay between HM silencing, telomeric silencing, and telomere length regulation. These findings suggest a model in which competition and multiple recruitment events modulate silencing and telomere length regulation.

  15. C-terminal truncation of RAP1 results in the deregulation of telomere size, stability, and function in Saccharomyces cerevisiae.

    PubMed Central

    Kyrion, G; Boakye, K A; Lustig, A J

    1992-01-01

    The Saccharomyces cerevisiae DNA-binding protein RAP1 is capable of binding in vitro to sequences from a wide variety of genomic loci, including upstream activating sequence elements, the HML and HMR silencer regions, and the poly(G1-3T) tracts of telomeres. Recent biochemical and genetic studies have suggested that RAP1 physically and functionally interacts with the yeast telomere. To further investigate the role of RAP1 at the telomere, we have identified and characterized three intragenic suppressors of a temperature-sensitive allele of RAP1, rap1-5. These telomere deficiency (rap1t) alleles confer several novel phenotypes. First, telomere tract size elongates to up to 4 kb greater than sizes of wild-type or rap1-5 telomeres. Second, telomeres are highly unstable and are subject to rapid, but reversible, deletion of part or all of the increase in telomeric tract length. Telomeric deletion does not require the RAD52 or RAD1 gene product. Third, chromosome loss and nondisjunction rates are elevated 15- to 30-fold above wild-type levels. Sequencing analysis has shown that each rap1t allele contains a nonsense mutation within a discrete region between amino acids 663 and 684. Mobility shift and Western immunoblot analyses indicate that each allele produces a truncated RAP1 protein, lacking the C-terminal 144 to 165 amino acids but capable of efficient DNA binding. These data suggest that RAP1 is a central regulator of both telomere and chromosome stability and define a C-terminal domain that, while dispensable for viability, is required for these telomeric functions. Images PMID:1406688

  16. Loss of the Rap1 effector RIAM results in leukocyte adhesion deficiency due to impaired β2 integrin function in mice.

    PubMed

    Klapproth, Sarah; Sperandio, Markus; Pinheiro, Elaine M; Prünster, Monika; Soehnlein, Oliver; Gertler, Frank B; Fässler, Reinhard; Moser, Markus

    2015-12-17

    Talin is an integrin adaptor, which controls integrin activity in all hematopoietic cells. How intracellular signals promote talin binding to the integrin tail leading to integrin activation is still poorly understood, especially in leukocytes. In vitro studies identified an integrin activation complex whose formation is initiated by the interaction of active, guanosine triphosphate (GTP)-bound Ras-related protein 1 (Rap1) with the adapter protein Rap1-GTP-interacting adapter molecule (RIAM) followed by the recruitment of talin to the plasma membrane. Unexpectedly, loss-of-function studies in mice have shown that the talin-activating role of RIAM is neither required for development nor for integrin activation in platelets. In this study, we show that leukocyte integrin activation critically depends on RIAM both in vitro and in vivo. RIAM deficiency results in a loss of β2 integrin activation in multiple leukocyte populations, impaired leukocyte adhesion to inflamed vessels, and accumulation in the circulation. Surprisingly, however, the major leukocyte β1 integrin family member, α4β1, was only partially affected by RIAM deficiency in leukocytes. Thus, although talin is an essential, shared regulator of all integrin classes expressed by leukocytes, we report that β2 and α4 integrins use different RIAM-dependent and -independent pathways to undergo activation by talin.

  17. Modulation of specific protein expression levels by PTEN: identification of AKAP121, DHFR, G3BP, Rap1, and RCC1 as potential targets of PTEN.

    PubMed

    Huang, Yanping; Wernyj, Roman P; Norton, Darrell D; Precht, Patricia; Seminario, Maria-Cristina; Wange, Ronald L

    2005-05-26

    The tumor suppressor PTEN is mutated in a high percentage of human cancers, and is implicated in pathways regulating cell growth, proliferation, survival, and migration. Despite significant advances, our understanding of its mechanisms of action remains incomplete. We have used a high-throughput proteomic immunoblotting approach to identify proteins whose expression levels are modulated by PTEN. Out of over 800 proteins screened, 22 proteins showed significant changes in expression. Five proteins that exhibited two-fold or greater changes in expression level were further characterized. AKAP121 and G3BP expression was reduced, while dihydrofolate reductase (DHFR), Rap1 and RCC1 expression was elevated in response to PTEN expression in a PTEN-null T-cell leukemia line. The phosphatase activity of PTEN was required for these effects. However, direct inhibition of PI-3 Kinase could mimic PTEN in modulating expression of DHFR, G3BP, Rap1 and RCC1, but not AKAP121. Real-time PCR showed that the effects of PTEN were primarily post-transcriptional, and would not have been revealed by mRNA-based screens. We conclude from these data that PTEN can modulate the expression level of a number of different proteins. The identified proteins have the potential to serve as previously unrecognized effectors of PTEN, and suggest the existence of additional complexity in the modes by which PTEN can regulate cellular biology.

  18. Mutations in the gene encoding KRIT1, a Krev-1/rap1a binding protein, cause cerebral cavernous malformations (CCM1).

    PubMed

    Sahoo, T; Johnson, E W; Thomas, J W; Kuehl, P M; Jones, T L; Dokken, C G; Touchman, J W; Gallione, C J; Lee-Lin, S Q; Kosofsky, B; Kurth, J H; Louis, D N; Mettler, G; Morrison, L; Gil-Nagel, A; Rich, S S; Zabramski, J M; Boguski, M S; Green, E D; Marchuk, D A

    1999-11-01

    Cerebral cavernous malformations (CCM) are congenital vascular anomalies of the brain that can cause significant neurological disabilities, including intractable seizures and hemorrhagic stroke. One locus for autosomal dominant CCM ( CCM1 ) maps to chromosome 7q21-q22. Recombination events in linked family members define a critical region of approximately 2 Mb and a shared disease haplotype associated with a presumed founder effect in families of Mexican-American descent points to a potentially smaller region of interest. Using a genomic sequence-based positional cloning strategy, we have identified KRIT1, encoding a protein that interacts with the Krev-1/rap1a tumor suppressor, as the CCM1 gene. Seven different KRIT1 mutations have been identified in 23 distinct CCM1 families. The identical mutation is present in 16 of 21 Mexican-American families analyzed, substantiating a founder effect in this population. Other Mexican-American and non-Hispanic Caucasian CCM1 kindreds harbor other KRIT1 mutations. Identification of a common Mexican-American mutation has potential clinical significance for presymptomatic diagnosis of CCM in this population. In addition, these data point to a key role for the Krev-1/rap1a signaling pathway in angiogenesis and cerebrovascular disease.

  19. Crucial Role of Rapgef2 and Rapgef6, a Family of Guanine Nucleotide Exchange Factors for Rap1 Small GTPase, in Formation of Apical Surface Adherens Junctions and Neural Progenitor Development in the Mouse Cerebral Cortex123

    PubMed Central

    Maeta, Kazuhiro; Edamatsu, Hironori; Nishihara, Kaori; Ikutomo, Junji; Bilasy, Shymaa E.

    2016-01-01

    Abstract Cerebral neocortex development in mammals requires highly orchestrated events involving proliferation, differentiation, and migration of neural progenitors and neurons. Rapgef2 and Rapgef6 constitute a unique family of guanine nucleotide exchange factors for Rap1 small GTPase, which is known to play crucial roles in migration of postmitotic neurons. We previously reported that conditional knockout of Rapgef2 in dorsal telencephalon (Rapgef2-cKO) resulted in the formation of an ectopic cortical mass (ECM) resembling that of subcortical band heterotopia. Here we show that double knockout of Rapgef6 in Rapgef2-cKO mice (Rapgef2/6-dKO) results in marked enlargement of the ECM. While Rapgef2-cKO affects late-born neurons only, Rapgef2/6-dKO affects both early-born and late-born neurons. The Rapgef2-cKO cortex at embryonic day (E) 15.5, and the Rapgef2/6-dKO cortex at E13.5 and E15.5 show disruption of the adherens junctions (AJs) on the apical surface, detachment of radial glial cells (RGCs) from the apical surface and disorganization of the radial glial fiber system, which are accompanied by aberrant distribution of RGCs and intermediate progenitors, normally located in the ventricular zone and the subventricular zone, respectively, over the entire cerebral cortex. Moreover, intrauterine transduction of Cre recombinase into the Rapgef2flox/flox brains also results in the apical surface AJ disruption and the RGC detachment from the apical surface, both of which are effectively suppressed by cotransduction of the constitutively active Rap1 mutant Rap1G12V. These results demonstrate a cell-autonomous role of the Rapgef2/6-Rap1 pathway in maintaining the apical surface AJ structures, which is necessary for the proper development of neural progenitor cells. PMID:27390776

  20. Social Context and Musical Content of Rap Music, 1979-1995

    ERIC Educational Resources Information Center

    Lena, Jennifer C.

    2006-01-01

    There is a link between the context of production and the content of rap music singles. This research finds that when independent labels owned most of the charted singles, lyrics emphasized features of the local environment and hostility to corporate music production and values. In contrast, the major-label dominated market featured lyrics…

  1. An Interview with Cathy Fowler about Sharing a Love of Reading through Book Raps.

    ERIC Educational Resources Information Center

    Strangman, Nicole

    2002-01-01

    Includes an interview with Cathy Fowler, a Year 7 teacher at Kawungan State School in Queensland, Australia. Explains that Cathy is a participant and coordinator of the extremely popular Harry Potter Book Rap, a guided Internet book discussion among students all over the world. Discusses how this activity fueled her students' love for reading. (PM)

  2. CDK5RAP2 stimulates microtubule nucleation by the gamma-tubulin ring complex.

    PubMed

    Choi, Yuk-Kwan; Liu, Pengfei; Sze, Siu Kwan; Dai, Chao; Qi, Robert Z

    2010-12-13

    CDK5RAP2 is a human microcephaly protein that contains a γ-tubulin complex (γ-TuC)-binding domain conserved in Drosophila melanogaster centrosomin and Schizosaccharomyces pombe Mto1p and Pcp1p, which are γ-TuC-tethering proteins. In this study, we show that this domain within CDK5RAP2 associates with the γ-tubulin ring complex (γ-TuRC) to stimulate its microtubule-nucleating activity and is therefore referred to as the γ-TuRC-mediated nucleation activator (γ-TuNA). γ-TuNA but not its γ-TuC-binding-deficient mutant stimulates microtubule nucleation by purified γ-TuRC in vitro and induces extensive, γ-TuRC-dependent nucleation of microtubules in a microtubule regrowth assay. γ-TuRC bound to γ-TuNA contains NME7, FAM128A/B, and actin in addition to γ-tubulin and GCP2-6. RNA interference-mediated depletion of CDK5RAP2 impairs both centrosomal and acentrosomal microtubule nucleation, although γ-TuRC assembly is unaffected. Collectively, these results suggest that the γ-TuNA found in CDK5RAP2 has regulatory functions in γ-TuRC-mediated microtubule nucleation. PMID:21135143

  3. RAP-011 improves erythropoiesis in zebrafish model of Diamond-Blackfan anemia through antagonizing lefty1.

    PubMed

    Ear, Jason; Huang, Haigen; Wilson, Tianna; Tehrani, Zahra; Lindgren, Anne; Sung, Victoria; Laadem, Abderrahmane; Daniel, Thomas O; Chopra, Rajesh; Lin, Shuo

    2015-08-13

    Diamond-Blackfan Anemia (DBA) is a bone marrow failure disorder characterized by low red blood cell count. Mutations in ribosomal protein genes have been identified in approximately half of all DBA cases. Corticosteriod therapy and bone marrow transplantation are common treatment options for patients; however, significant risks and complications are associated with these treatment options. Therefore, novel therapeutic approaches are needed for treating DBA. Sotatercept (ACE-011, and its murine ortholog RAP-011) acts as an activin receptor type IIA ligand trap, increasing hemoglobin and hematocrit in pharmacologic models, in healthy volunteers, and in patients with β-thalassemia, by expanding late-stage erythroblasts through a mechanism distinct from erythropoietin. Here, we evaluated the effects of RAP-011 in zebrafish models of RPL11 ribosome deficiency. Treatment with RAP-011 dramatically restored hemoglobin levels caused by ribosome stress. In zebrafish embryos, RAP-011 likely stimulates erythropoietic activity by sequestering lefty1 from erythroid cells. These findings identify lefty1 as a signaling component in the development of erythroid cells and rationalize the use of sotatercept in DBA patients. PMID:26109203

  4. RAP-011 improves erythropoiesis in zebrafish model of Diamond-Blackfan anemia through antagonizing lefty1.

    PubMed

    Ear, Jason; Huang, Haigen; Wilson, Tianna; Tehrani, Zahra; Lindgren, Anne; Sung, Victoria; Laadem, Abderrahmane; Daniel, Thomas O; Chopra, Rajesh; Lin, Shuo

    2015-08-13

    Diamond-Blackfan Anemia (DBA) is a bone marrow failure disorder characterized by low red blood cell count. Mutations in ribosomal protein genes have been identified in approximately half of all DBA cases. Corticosteriod therapy and bone marrow transplantation are common treatment options for patients; however, significant risks and complications are associated with these treatment options. Therefore, novel therapeutic approaches are needed for treating DBA. Sotatercept (ACE-011, and its murine ortholog RAP-011) acts as an activin receptor type IIA ligand trap, increasing hemoglobin and hematocrit in pharmacologic models, in healthy volunteers, and in patients with β-thalassemia, by expanding late-stage erythroblasts through a mechanism distinct from erythropoietin. Here, we evaluated the effects of RAP-011 in zebrafish models of RPL11 ribosome deficiency. Treatment with RAP-011 dramatically restored hemoglobin levels caused by ribosome stress. In zebrafish embryos, RAP-011 likely stimulates erythropoietic activity by sequestering lefty1 from erythroid cells. These findings identify lefty1 as a signaling component in the development of erythroid cells and rationalize the use of sotatercept in DBA patients.

  5. A higher-order entity formed by the flexible assembly of RAP1 with TRF2

    PubMed Central

    Gaullier, Guillaume; Miron, Simona; Pisano, Sabrina; Buisson, Rémi; Le Bihan, Yann-Vaï; Tellier-Lebègue, Carine; Messaoud, Wala; Roblin, Pierre; Guimarães, Beatriz G.; Thai, Robert; Giraud-Panis, Marie-Josèphe; Gilson, Eric; Le Du, Marie-Hélène

    2016-01-01

    Telomere integrity is essential to maintain genome stability, and telomeric dysfunctions are associated with cancer and aging pathologies. In human, the shelterin complex binds TTAGGG DNA repeats and provides capping to chromosome ends. Within shelterin, RAP1 is recruited through its interaction with TRF2, and TRF2 is required for telomere protection through a network of nucleic acid and protein interactions. RAP1 is one of the most conserved shelterin proteins although one unresolved question is how its interaction may influence TRF2 properties and regulate its capacity to bind multiple proteins. Through a combination of biochemical, biophysical and structural approaches, we unveiled a unique mode of assembly between RAP1 and TRF2. The complete interaction scheme between the full-length proteins involves a complex biphasic interaction of RAP1 that directly affects the binding properties of the assembly. These results reveal how a non-DNA binding protein can influence the properties of a DNA-binding partner by mutual conformational adjustments. PMID:26748096

  6. Rapping in Catalan in Class and the Empowerment of the Learner

    ERIC Educational Resources Information Center

    Aliagas, Cristina; Fernández, Júlia-Alba; Llonch, Pau

    2016-01-01

    Despite the well-known educational possibilities afforded by "Rhythm And Poetry" (RAP) for the development of musical, lyrical and critical skills [Morrell, E., & Duncan-Andrade, J. M. R. (2002). Promoting Academic Literacy with Urban Youth through Engaging Hip-hop Culture. "The English Journal," 91(6), 88-92. Retrieved…

  7. Operation Storefront. A Computer-Assisted Instruction Component of the Reading Assistance Program (RAP).

    ERIC Educational Resources Information Center

    Pasco County District School Board, Land O'Lakes, FL.

    This document contains teacher's materials that accompany Operation Storefront, the computer-assisted instruction package used in the Reading Assistance Program (RAP) that serves nonreading adults in Pasco County, Florida, who are 16 years or older and not in school. The target population for Operation Storefront is adults reading at or below the…

  8. The Racial Attitudes and Perceptions Survey (RAPS). Final Report (Mar 73-Mar 74). Technical Paper 338.

    ERIC Educational Resources Information Center

    Hiett, Robert L.; And Others

    The Racial Attitudes and Perceptions Survey (RAPS) was developed to obtain information from black and white military personnel, including reports of the frequencies of specific discriminatory behaviors and the tension levels associated with each race. The instrument was evaluated in terms of its construct validity and its reliability. Attitudes…

  9. "My Mom and Her Boyfriend Fuss": A Five-Minute RAP

    ERIC Educational Resources Information Center

    Laursen, Erik K.; Whindleton, Kendra

    2012-01-01

    Unresolved issues from home often spill over into behavior problems at school. This article discusses the five-minute RAP techniques: (1) connect; (2) clarify; and (3) restore. The authors present a case study wherein this brief restorative intervention had been a successful alternative to punishment and exclusion.

  10. Illuminating Chaucer through Poetry, Manuscript Illuminations, and a Critical Rap Album

    ERIC Educational Resources Information Center

    Lynch, Tom Liam

    2007-01-01

    Drawing connections between Chaucer, Eminem, and social issues, New York City high school teacher Tom Liam Lynch helped students become familiar with "The Canterbury Tales." Students wrote poems of rhymed couplets about today's social and political issues, created illuminated manuscripts, and recorded a rap CD. A book and album were published for…

  11. Relationships between Exposure to Rap Music Videos and Attitudes toward Relationships among African American Youth

    ERIC Educational Resources Information Center

    Bryant, Yaphet

    2008-01-01

    The purpose of the study is to (a) predict adversarial attitudes toward male-female relationships and (b) explore the relationships between traditional agents of socialization and personal acceptance of negative images in rap videos by African American adolescents. Participants completed psychosocial measures, viewed videos, and completed surveys…

  12. The RAP: A Recreational Activities Project, Academic Service-Learning Course and Qualitative Research Study

    ERIC Educational Resources Information Center

    Parker, Kathlyn

    2009-01-01

    The author (a university instructor) and her community partner (a public school teacher) have collaborated in teaching an academic service-learning course in special education. This collaboration, the RAP (recreational activities project), was completed by university undergraduate students and young adults with cognitive impairment and/or…

  13. Using Rap and Jamaican Dance Hall Music in the Secondary Music Classroom

    ERIC Educational Resources Information Center

    Minott, Mark

    2008-01-01

    This article reports on a study carried out in a secondary school in the Island of Jamaica. One grade 7 class (n = 20) and one grade 9 class (n = 23) were taught a six-week unit of lessons aimed at facilitating student listening, performing and composing. Rap and Jamaican dance hall music were used as the stimulus for students' rhythmic…

  14. Impulse Control Rap: "We Got a Skill to Help You Chill."

    ERIC Educational Resources Information Center

    Tyrone; Hall, Chris A.; Hill, John W.

    1998-01-01

    Describes how a 16-year-old African-American male, Tyrone, excelled in a mental-health day-treatment facility in which he was enrolled. Tyrone used the skills taught within the replacement-skills curriculum at the facility to compose a rap song that reminded him and his classmates how to react when faced with a difficult situation. (MKA)

  15. Seeing White through Rap: A Classroom Exercise for Examining Race Using a Hip-Hop Video

    ERIC Educational Resources Information Center

    Stein, Robert

    2011-01-01

    When discussing race in the classroom, getting students--including, importantly, white students--to see that all people have racial identities can be a challenge. This classroom exercise employs a rap video as a tool for helping to reveal white as a racial identity and for exploring the concept of white privilege--the idea that economic, social,…

  16. Reinforcing Alcohol Prevention (RAP) Program: A Secondary School Curriculum to Combat Underage Drinking and Impaired Driving

    ERIC Educational Resources Information Center

    Will, Kelli England; Sabo, Cynthia Shier

    2010-01-01

    The Reinforcing Alcohol Prevention (RAP) Program is an alcohol prevention curriculum developed in partnership with secondary schools to serve their need for a brief, evidence-based, and straightforward program that aligned with state learning objectives. Program components included an educational lesson, video, and interactive activities delivered…

  17. The Relationship between Heavy Metal and Rap Music and Adolescent Turmoil: Real or Artifact?

    ERIC Educational Resources Information Center

    Took, Kevin J.; Weiss, David S.

    1994-01-01

    Investigated association between 87 adolescents' music preferences and psychosocial turmoil. Adolescents who preferred heavy metal and rap music had higher incidence of below-average school grades, school behavior problems, sexual activity, drug and alcohol use, and arrests. When gender was controlled, only below-average school grades and history…

  18. Strong conservation of rhoptry-associated-protein-1 (RAP-1) locus organization and sequence among Babesia isolates infecting sheep from China (Babesia motasi-like phylogenetic group).

    PubMed

    Niu, Qingli; Valentin, Charlotte; Bonsergent, Claire; Malandrin, Laurence

    2014-12-01

    Rhoptry-associated-protein 1 (RAP-1) is considered as a potential vaccine candidate due to its involvement in red blood cell invasion by parasites in the genus Babesia. We examined its value as a vaccine candidate by studying RAP-1 conservation in isolates of Babesia sp. BQ1 Ningxian, Babesia sp. Tianzhu and Babesia sp. Hebei, responsible for ovine babesiosis in different regions of China. The rap-1 locus in these isolates has very similar features to those described for Babesia sp. BQ1 Lintan, another Chinese isolate also in the B. motasi-like phylogenetic group, namely the presence of three types of rap-1 genes (rap-1a, rap-1b and rap-1c), multiple conserved rap-1b copies (5) interspaced with more or less variable rap-1a copies (6), and the 3' localization of one rap-1c. The isolates Babesia sp. Tianzhu, Babesia sp. BQ1 Lintan and Ningxian were almost identical (average nucleotide identity of 99.9%) over a putative locus of about 31 Kb, including the intergenic regions. Babesia sp. Hebei showed a similar locus organization but differed in the rap-1 locus sequence, for each gene and intergenic region, with an average nucleotide identity of 78%. Our results are in agreement with 18S rDNA phylogenetic studies performed on these isolates. However, in extremely closely related isolates the rap-1 locus seems more conserved (99.9%) than the 18S rDNA (98.7%), whereas in still closely related isolates the identities are much lower (78%) compared with the 18S rDNA (97.7%). The particularities of the rap-1 locus in terms of evolution, phylogeny, diagnosis and vaccine development are discussed.

  19. Representative Agricultural Pathways and Climate Impact Assessment for Pacific Northwest Agricultural Systems

    NASA Astrophysics Data System (ADS)

    MU, J.; Antle, J. M.; Zhang, H.; Capalbo, S. M.; Eigenbrode, S.; Kruger, C.; Stockle, C.; Wolfhorst, J. D.

    2013-12-01

    Representative Agricultural Pathways (RAPs) are projections of plausible future biophysical and socio-economic conditions used to carry out climate impact assessments for agriculture. The development of RAPs iss motivated by the fact that the various global and regional models used for agricultural climate change impact assessment have been implemented with individualized scenarios using various data and model structures, often without transparent documentation or public availability. These practices have hampered attempts at model inter-comparison, improvement, and synthesis of model results across studies. This paper aims to (1) present RAPs developed for the principal wheat-producing region of the Pacific Northwest, and to (2) combine these RAPs with downscaled climate data, crop model simulations and economic model simulations to assess climate change impacts on winter wheat production and farm income. This research was carried out as part of a project funded by the USDA known as the Regional Approaches to Climate Change in the Pacific Northwest (REACCH). The REACCH study region encompasses the major winter wheat production area in Pacific Northwest and preliminary research shows that farmers producing winter wheat could benefit from future climate change. However, the future world is uncertain in many dimensions, including commodity and input prices, production technology, and policies, as well as increased probability of disturbances (pests and diseases) associated with a changing climate. Many of these factors cannot be modeled, so they are represented in the regional RAPS. The regional RAPS are linked to global agricultural and shared social-economic pathways, and used along with climate change projections to simulate future outcomes for the wheat-based farms in the REACCH region.

  20. Crystal structure of the C-terminal domain of the RAP74 subunit of human transcription factor IIF

    SciTech Connect

    Kamada, Katsuhiko; De Angelis, Jacqueline; Roeder, Robert G.; Burley, Stephen K.

    2012-12-13

    The x-ray structure of a C-terminal fragment of the RAP74 subunit of human transcription factor (TF) IIF has been determined at 1.02-{angstrom} resolution. The {alpha}/{beta} structure is strikingly similar to the globular domain of linker histone H5 and the DNA-binding domain of hepatocyte nuclear factor 3{gamma} (HNF-3{gamma}), making it a winged-helix protein. The surface electrostatic properties of this compact domain differ significantly from those of bona fide winged-helix transcription factors (HNF-3{gamma} and RFX1) and from the winged-helix domains found within the RAP30 subunit of TFIIF and the {beta} subunit of TFIIE. RAP74 has been shown to interact with the TFIIF-associated C-terminal domain phosphatase FCP1, and a putative phosphatase binding site has been identified within the RAP74 winged-helix domain.

  1. RapTOR: Automated sequencing library preparation and suppression for rapid pathogen characterization ( 7th Annual SFAF Meeting, 2012)

    ScienceCinema

    Lane, Todd [SNL

    2016-07-12

    Todd Lane on "RapTOR: Automated sequencing library preparation and suppression for rapid pathogen characterization" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  2. RapTOR: Automated sequencing library preparation and suppression for rapid pathogen characterization ( 7th Annual SFAF Meeting, 2012)

    SciTech Connect

    Lane, Todd

    2012-06-01

    Todd Lane on "RapTOR: Automated sequencing library preparation and suppression for rapid pathogen characterization" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  3. Effects of using nursing home residents to serve as group activity leaders: lessons learned from the RAP project.

    PubMed

    Skrajner, Michael J; Haberman, Jessica L; Camp, Cameron J; Tusick, Melanie; Frentiu, Cristina; Gorzelle, Gregg

    2014-03-01

    Previous research has demonstrated that persons with early to moderate stage dementia are capable of leading small group activities for persons with more advanced dementia. In this study, we built upon this previous work by training residents in long-term care facilities to fill the role of group activity leaders using a Resident-Assisted Programming (RAP) training regimen. There were two stages to the program. In the first stage, RAP training was provided by researchers. In the second stage, RAP training was provided to residents by activities staff members of long-term care facilities who had been trained by researchers. We examine the effects of RAP implemented by researchers and by activities staff member on long-term care resident with dementia who took part in these RAP activities. We also examined effects produced by two types of small group activities: two Montessori-based activities and an activity which focuses on persons with more advanced dementia, based on the work of Jitka Zgola. Results demonstrate that levels of positive engagement seen in players during RAP (resident-led activities) were typically higher than those observed during standard activities programming led by site staff. In general, Montessori-Based Dementia Programming® produced more constructive engagement than Zgola-based programming (ZBP), though ZBP did increase a positive form of engagement involving observing activities with interest. In addition, RAP implemented by activities staff members produced effects that were, on the whole, similar to those produced when RAP was implemented by researchers. Implications of these findings for providing meaningful social roles for persons with dementia residing in long-term care, and suggestions for further research in this area, are discussed.

  4. Activation of brain B-Raf protein kinase by Rap1B small GTP-binding protein.

    PubMed

    Ohtsuka, T; Shimizu, K; Yamamori, B; Kuroda, S; Takai, Y

    1996-01-19

    Rap1 small GTP-binding protein has the same amino acid sequence at its effector domain as that of Ras. Rap1 has been shown to antagonize the Ras functions, such as the Ras-induced transformation of NIH 3T3 cells and the Ras-induced activation of the c-Raf-1 protein kinase-dependent mitogen-activated protein (MAP) kinase cascade in Rat-1 cells, whereas we have shown that Rap1 as well as Ras stimulates DNA synthesis in Swiss 3T3 cells. We have established a cell-free assay system in which Ras activates bovine brain B-Raf protein kinase. Here we have used this assay system and examined the effect of Rap1 on the B-Raf activity to phosphorylate recombinant MAP kinase kinase (MEK). Recombinant Rap1B stimulated the activity of B-Raf, which was partially purified from bovine brain and immunoprecipitated by an anti-B-Raf antibody. The GTP-bound form was active, but the GDP-bound form was inactive. The fully post-translationally lipid-modified form was active, but the unmodified form was nearly inactive. The maximum B-Raf activity stimulated by Rap1B was nearly the same as that stimulated by Ki-Ras. Rap1B enhanced the Ki-Ras-stimulated B-Raf activity in an additive manner. These results indicate that not only Ras but also Rap1 is involved in the activation of the B-Raf-dependent MAP kinase cascade.

  5. Quantitative Assessment of RNA-Protein Interactions with High Throughput Sequencing - RNA Affinity Profiling (HiTS-RAP)

    PubMed Central

    Ozer, Abdullah; Tome, Jacob M.; Friedman, Robin C.; Gheba, Dan; Schroth, Gary P.; Lis, John T.

    2016-01-01

    Because RNA-protein interactions play a central role in a wide-array of biological processes, methods that enable a quantitative assessment of these interactions in a high-throughput manner are in great demand. Recently, we developed the High Throughput Sequencing-RNA Affinity Profiling (HiTS-RAP) assay, which couples sequencing on an Illumina GAIIx with the quantitative assessment of one or several proteins’ interactions with millions of different RNAs in a single experiment. We have successfully used HiTS-RAP to analyze interactions of EGFP and NELF-E proteins with their corresponding canonical and mutant RNA aptamers. Here, we provide a detailed protocol for HiTS-RAP, which can be completed in about a month (8 days hands-on time) including the preparation and testing of recombinant proteins and DNA templates, clustering DNA templates on a flowcell, high-throughput sequencing and protein binding with GAIIx, and finally data analysis. We also highlight aspects of HiTS-RAP that can be further improved and points of comparison between HiTS-RAP and two other recently developed methods, RNA-MaP and RBNS. A successful HiTS-RAP experiment provides the sequence and binding curves for approximately 200 million RNAs in a single experiment. PMID:26182240

  6. Structural basis for activation and non-canonical catalysis of the Rap GTPase activating protein domain of plexin.

    PubMed

    Wang, Yuxiao; Pascoe, Heath G; Brautigam, Chad A; He, Huawei; Zhang, Xuewu

    2013-10-01

    Plexins are cell surface receptors that bind semaphorins and transduce signals for regulating neuronal axon guidance and other processes. Plexin signaling depends on their cytoplasmic GTPase activating protein (GAP) domain, which specifically inactivates the Ras homolog Rap through an ill-defined non-canonical catalytic mechanism. The plexin GAP is activated by semaphorin-induced dimerization, the structural basis for which remained unknown. Here we present the crystal structures of the active dimer of zebrafish PlexinC1 cytoplasmic region in the apo state and in complex with Rap. The structures show that the dimerization induces a large-scale conformational change in plexin, which opens the GAP active site to allow Rap binding. Plexin stabilizes the switch II region of Rap in an unprecedented conformation, bringing Gln63 in Rap into the active site for catalyzing GTP hydrolysis. The structures also explain the unique Rap-specificity of plexins. Mutational analyses support that these mechanisms underlie plexin activation and signaling. DOI:http://dx.doi.org/10.7554/eLife.01279.001.

  7. Cdk5rap2 exposes the centrosomal root of microcephaly syndromes.

    PubMed

    Megraw, Timothy L; Sharkey, James T; Nowakowski, Richard S

    2011-08-01

    Autosomal recessive primary microcephaly (MCPH) is characterized by small brain size as a result of deficient neuron production in the developing cerebral cortex. Although MCPH is a rare disease, the questions surrounding its etiology strike at the core of stem cell biology. The seven genes implicated in MCPH all encode centrosomal proteins and disruption of the MCPH gene Cdk5rap2 in mice revealed its role in neural progenitor proliferation and in maintaining normal centriole replication control. We discuss here the impact that centrosome regulation has upon neural progenitors in the developing brain. We integrate the impact of centriole replication defects with the functions of Cdk5rap2 and other MCPH proteins, propose mechanisms for progenitor loss in MCPH, and discuss links to two other microcephaly syndromes. PMID:21632253

  8. RAP80 Targets BRCA1 to Specific Ubiquitin Structures at DNA Damage Sites

    PubMed Central

    Sobhian, Bijan; Shao, Genze; Lilli, Dana R.; Culhane, Aedín C.; Moreau, Lisa A.; Xia, Bing; Livingston, David M.; Greenberg, Roger A.

    2009-01-01

    Mutations affecting the BRCT domains of the breast cancer–associated tumor suppressor BRCA1 disrupt the recruitment of this protein to DNA double-strand breaks (DSBs). The molecular structures at DSBs recognized by BRCA1 are presently unknown. We report the interaction of the BRCA1 BRCT domain with RAP80, a ubiquitin-binding protein. RAP80 targets a complex containing the BRCA1-BARD1 (BRCA1-associated ring domain protein 1) E3 ligase and the deubiquitinating enzyme (DUB) BRCC36 to MDC1-γH2AX–dependent lysine6- and lysine63-linked ubiquitin polymers at DSBs. These events are required for cell cycle checkpoint and repair responses to ionizing radiation, implicating ubiquitin chain recognition and turnover in the BRCA1-mediated repair of DSBs. PMID:17525341

  9. Photometric redshift estimation based on data mining with PhotoRApToR

    NASA Astrophysics Data System (ADS)

    Cavuoti, S.; Brescia, M.; De Stefano, V.; Longo, G.

    2015-03-01

    Photometric redshifts (photo-z) are crucial to the scientific exploitation of modern panchromatic digital surveys. In this paper we present PhotoRApToR (Photometric Research Application To Redshift): a Java/C ++ based desktop application capable to solve non-linear regression and multi-variate classification problems, in particular specialized for photo-z estimation. It embeds a machine learning algorithm, namely a multi-layer neural network trained by the Quasi Newton learning rule, and special tools dedicated to pre- and post-processing data. PhotoRApToR has been successfully tested on several scientific cases. The application is available for free download from the DAME Program web site.

  10. Rho and Rap guanosine triphosphatase signaling in B cells and chronic lymphocytic leukemia.

    PubMed

    Mele, Silvia; Devereux, Stephen; Ridley, Anne J

    2014-09-01

    Chronic lymphocytic leukemia (CLL) cells proliferate predominantly in niches in the lymph nodes, where signaling from the B cell receptor (BCR) and the surrounding microenvironment are critical for disease progression. In addition, leukemic cells traffic constantly from the bloodstream into the lymph nodes, migrate within lymphatic tissues and egress back to the bloodstream. These processes are driven by chemokines and their receptors, and depend on changes in cell migration and integrin-mediated adhesion. Here we describe how Rho and Rap guanosine triphosphatases (GTPases) contribute to both BCR signaling and chemokine receptor signaling, particularly by regulating cytoskeletal dynamics and integrin activity. We propose that new inhibitors of BCR-activated kinases are likely to affect CLL cell trafficking via Rho and Rap GTPases, and that upstream regulators or downstream effectors could be good targets for therapeutic intervention in CLL.

  11. A centrosomal mechanism involving CDK5RAP2 and CENPJ controls brain size.

    PubMed

    Bond, Jacquelyn; Roberts, Emma; Springell, Kelly; Lizarraga, Sofia B; Lizarraga, Sophia; Scott, Sheila; Higgins, Julie; Hampshire, Daniel J; Morrison, Ewan E; Leal, Gabriella F; Silva, Elias O; Costa, Suzana M R; Baralle, Diana; Raponi, Michela; Karbani, Gulshan; Rashid, Yasmin; Jafri, Hussain; Bennett, Christopher; Corry, Peter; Walsh, Christopher A; Woods, C Geoffrey

    2005-04-01

    Autosomal recessive primary microcephaly is a potential model in which to research genes involved in human brain growth. We show that two forms of the disorder result from homozygous mutations in the genes CDK5RAP2 and CENPJ. We found neuroepithelial expression of the genes during prenatal neurogenesis and protein localization to the spindle poles of mitotic cells, suggesting that a centrosomal mechanism controls neuron number in the developing mammalian brain. PMID:15793586

  12. Rice Annotation Project Database (RAP-DB): an integrative and interactive database for rice genomics.

    PubMed

    Sakai, Hiroaki; Lee, Sung Shin; Tanaka, Tsuyoshi; Numa, Hisataka; Kim, Jungsok; Kawahara, Yoshihiro; Wakimoto, Hironobu; Yang, Ching-chia; Iwamoto, Masao; Abe, Takashi; Yamada, Yuko; Muto, Akira; Inokuchi, Hachiro; Ikemura, Toshimichi; Matsumoto, Takashi; Sasaki, Takuji; Itoh, Takeshi

    2013-02-01

    The Rice Annotation Project Database (RAP-DB, http://rapdb.dna.affrc.go.jp/) has been providing a comprehensive set of gene annotations for the genome sequence of rice, Oryza sativa (japonica group) cv. Nipponbare. Since the first release in 2005, RAP-DB has been updated several times along with the genome assembly updates. Here, we present our newest RAP-DB based on the latest genome assembly, Os-Nipponbare-Reference-IRGSP-1.0 (IRGSP-1.0), which was released in 2011. We detected 37,869 loci by mapping transcript and protein sequences of 150 monocot species. To provide plant researchers with highly reliable and up to date rice gene annotations, we have been incorporating literature-based manually curated data, and 1,626 loci currently incorporate literature-based annotation data, including commonly used gene names or gene symbols. Transcriptional activities are shown at the nucleotide level by mapping RNA-Seq reads derived from 27 samples. We also mapped the Illumina reads of a Japanese leading japonica cultivar, Koshihikari, and a Chinese indica cultivar, Guangluai-4, to the genome and show alignments together with the single nucleotide polymorphisms (SNPs) and gene functional annotations through a newly developed browser, Short-Read Assembly Browser (S-RAB). We have developed two satellite databases, Plant Gene Family Database (PGFD) and Integrative Database of Cereal Gene Phylogeny (IDCGP), which display gene family and homologous gene relationships among diverse plant species. RAP-DB and the satellite databases offer simple and user-friendly web interfaces, enabling plant and genome researchers to access the data easily and facilitating a broad range of plant research topics.

  13. Magnetic control in the RAP-200K-20 x-ray equipment

    SciTech Connect

    Gusev, E.A.; Drankov, V.P.; Naboishchikov, V.D.

    1989-03-01

    A description is given of the RAP-200K-20 cable-connected x-ray equipment, where a three-phase EHT transformer with magnetic control is used in the main circuit. The apparatus is compared with the best foreign competition. The circuit has an advantage over a pulse regulator in that the overvoltage level is low; there is also no interference and the efficiency is higher. All these advantages improve the performance and reliability in TV and fluorescent monitoring.

  14. Real-time Aerosol Forecasting over North America using RAP-Chem and the GSI.

    NASA Astrophysics Data System (ADS)

    Pagowski, M.

    2015-12-01

    RAP-Chem is an implementation of WRF-Chem meteorology-chemistry model that is run daily at NOAA/ESRL over continental domain for air-quality forecasting. The chemical forecasts are combined with observations of species using three-dimensional variational data assimilation procedure implemented in the Gridpoint Statistical Interpolation (GSI). In the presentation we detail the method of the assimilation and show verification statistics of the model performance.

  15. Dab2IP Regulates Neuronal Positioning, Rap1 Activity and Integrin Signaling in the Developing Cortex.

    PubMed

    Qiao, Shuhong; Homayouni, Ramin

    2015-01-01

    Dab2IP (DOC-2/DAB2 interacting protein) is a GTPase-activating protein which is involved in various aspects of brain development in addition to its roles in tumor formation and apoptosis in other systems. In this study, we carefully examined the expression profile of Dab2IP and investigated its physiological role during brain development using a Dab2IP-knockdown (KD) mouse model created by retroviral insertion of a LacZ-encoding gene-trapping cassette. LacZ staining revealed that Dab2IP is expressed in the ventricular zone as well as the cortical plate and the intermediate zone. Immunohistochemical analysis showed that Dab2IP protein is localized in the leading process and proximal cytoplasmic regions of migrating neurons in the intermediate zone. Bromodeoxyuridine birth dating experiments in combination with immunohistochemical analysis using layer-specific markers showed that Dab2IP is important for proper positioning of a subset of layer II-IV neurons in the developing cortex. Notably, neuronal migration was not completely disrupted in the cerebral cortex of Dab2IP-KD mice and disruption of migration was not strictly layer specific. Previously, we found that Dab2IP regulates multipolar transition in cortical neurons. Others have shown that Rap1 regulates the transition from multipolar to bipolar morphology in migrating postmitotic neurons through N-cadherin signaling and somal translocation in the superficial layer of the cortical plate through integrin signaling. Therefore, we examined whether Rap1 and integrin signaling were affected in Dab2IP-KD brains. We found that Dab2IP-KD resulted in higher levels of activated Rap1 and integrin in the developing cortex. Taken together, our results suggest that Dab2IP plays an important role in the migration and positioning of a subpopulation of later-born (layers II-IV) neurons, likely through the regulation of Rap1 and integrin signaling. PMID:25721469

  16. Dab2IP Regulates Neuronal Positioning, Rap1 Activity and Integrin Signaling in the Developing Cortex.

    PubMed

    Qiao, Shuhong; Homayouni, Ramin

    2015-01-01

    Dab2IP (DOC-2/DAB2 interacting protein) is a GTPase-activating protein which is involved in various aspects of brain development in addition to its roles in tumor formation and apoptosis in other systems. In this study, we carefully examined the expression profile of Dab2IP and investigated its physiological role during brain development using a Dab2IP-knockdown (KD) mouse model created by retroviral insertion of a LacZ-encoding gene-trapping cassette. LacZ staining revealed that Dab2IP is expressed in the ventricular zone as well as the cortical plate and the intermediate zone. Immunohistochemical analysis showed that Dab2IP protein is localized in the leading process and proximal cytoplasmic regions of migrating neurons in the intermediate zone. Bromodeoxyuridine birth dating experiments in combination with immunohistochemical analysis using layer-specific markers showed that Dab2IP is important for proper positioning of a subset of layer II-IV neurons in the developing cortex. Notably, neuronal migration was not completely disrupted in the cerebral cortex of Dab2IP-KD mice and disruption of migration was not strictly layer specific. Previously, we found that Dab2IP regulates multipolar transition in cortical neurons. Others have shown that Rap1 regulates the transition from multipolar to bipolar morphology in migrating postmitotic neurons through N-cadherin signaling and somal translocation in the superficial layer of the cortical plate through integrin signaling. Therefore, we examined whether Rap1 and integrin signaling were affected in Dab2IP-KD brains. We found that Dab2IP-KD resulted in higher levels of activated Rap1 and integrin in the developing cortex. Taken together, our results suggest that Dab2IP plays an important role in the migration and positioning of a subpopulation of later-born (layers II-IV) neurons, likely through the regulation of Rap1 and integrin signaling.

  17. Molecular evolution of the brain size regulator genes CDK5RAP2 and CENPJ.

    PubMed

    Evans, Patrick D; Vallender, Eric J; Lahn, Bruce T

    2006-06-21

    Primary microcephaly is a developmental defect of the brain characterized by severely reduced brain size but an absence of other overt abnormalities. Mutations in several loci have been linked to primary microcephaly. The underlying genes for two of these were recently identified as CDK5RAP2 and CENPJ. Here, we focus on CDK5RAP2 and show that the protein evolutionary rate of this gene is significantly higher in primates than rodents or carnivores. We further show that the evolutionary rate within primates is particularly high in the human and chimpanzee terminal branches. Thus, the pattern of molecular evolution seen in CDK5RAP2 appears to parallel, at least approximately, that seen in two other previously identified primary microcephaly genes, microcephalin and ASPM. We also briefly discuss CENPJ, which similarly exhibits higher rate of protein evolution in primates as compared to rodents and carnivores. Together, the evolutionary patterns of all four presently known primary microcephaly genes are consistent with the hypothesis that genes regulating brain size during development might also play a role in brain evolution in primates and especially humans. PMID:16631324

  18. Re-active Passive (RAP) Devices for Control of Noise Transmission through a Panel

    NASA Technical Reports Server (NTRS)

    Carneal, James P.; Giovanardi, Marco; Fuller, Chris R.; Palumbo, Daniel L.

    2008-01-01

    Re-Active Passive (RAP) devices have been developed to control low frequency (<1000 Hz) noise transmission through a panel. These devices use a combination of active, re-active, and passive technologies packaged into a single unit to control a broad frequency range utilizing the strength of each technology over its best suited frequency range. The RAP device uses passive constrained layer damping to cover the relatively high frequency range (>200 Hz), reactive distributed vibration absorber) to cover the medium frequency range (75 to 250 Hz), and active control for controlling low frequencies (<200 Hz). The device was applied to control noise transmission through a panel mounted in a transmission loss test facility. Experimental results are presented for the bare panel, and combinations of passive treatment, reactive treatment, and active control. Results indicate that three RAP devices were able to increase the overall broadband (15-1000 Hz) transmission loss by 9.4 dB. These three devices added a total of 285 grams to the panel mass of 6.0 kg, or approximately 5%, not including control electronics.

  19. The High Resolution Accelerometer Package (HiRAP) flight experiment summary for the first 10 flights

    NASA Technical Reports Server (NTRS)

    Blanchard, Robert C.; Larman, K. T.; Barrett, M.

    1992-01-01

    The High Resolution Accelerometer Package (HiRAP) instrument is a triaxial, orthogonal system of gas damped accelerometers with a resolution of 1 x 10(exp -6) g (1 micro-g). The purpose of HiRAP is to measure the low frequency component of the total acceleration along the orbiter vehicle (OV) body axes while the OV descends through the rarefied flow flight regime. Two HiRAP instruments have flown on a total of 10 Space Transport System (STS) missions. The aerodynamic component of the acceleration measurements was separated from the total acceleration. Instrument bias and orbiter mechanical system acceleration effects were incorporated into one bulk bias. The bulk bias was subtracted from the acceleration measurements to produce aerodynamic descent data sets for all 10 flights. The aerodynamic acceleration data sets were input to an aerodynamic coefficient model. The aerodynamic acceleration data and coefficient model were used to estimate the atmospheric density for the altitude range of 140 to 60 km and a downrange distance of 600 km. For 8 of 10 flights results from this model agree with expected results. For the results that do not agree with expected results, a variety of error sources have been explored.

  20. Epac-Rap signaling reduces oxidative stress in the tubular epithelium.

    PubMed

    Stokman, Geurt; Qin, Yu; Booij, Tijmen H; Ramaiahgari, Sreenivasa; Lacombe, Marie; Dolman, M Emmy M; van Dorenmalen, Kim M A; Teske, Gwendoline J D; Florquin, Sandrine; Schwede, Frank; van de Water, Bob; Kok, Robbert J; Price, Leo S

    2014-07-01

    Activation of Rap1 by exchange protein activated by cAMP (Epac) promotes cell adhesion and actin cytoskeletal polarization. Pharmacologic activation of Epac-Rap signaling by the Epac-selective cAMP analog 8-pCPT-2'-O-Me-cAMP during ischemia-reperfusion (IR) injury reduces renal failure and application of 8-pCPT-2'-O-Me-cAMP promotes renal cell survival during exposure to the nephrotoxicant cisplatin. Here, we found that activation of Epac by 8-pCPT-2'-O-Me-cAMP reduced production of reactive oxygen species during reoxygenation after hypoxia by decreasing mitochondrial superoxide production. Epac activation prevented disruption of tubular morphology during diethyl maleate-induced oxidative stress in an organotypic three-dimensional culture assay. In vivo renal targeting of 8-pCPT-2'-O-Me-cAMP to proximal tubules using a kidney-selective drug carrier approach resulted in prolonged activation of Rap1 compared with nonconjugated 8-pCPT-2'-O-Me-cAMP. Activation of Epac reduced antioxidant signaling during IR injury and prevented tubular epithelial injury, apoptosis, and renal failure. Our data suggest that Epac1 decreases reactive oxygen species production by preventing mitochondrial superoxide formation during IR injury, thus limiting the degree of oxidative stress. These findings indicate a new role for activation of Epac as a therapeutic application in renal injury associated with oxidative stress.

  1. Redundancy in regulation of chondrogenesis in MIA/CD-RAP-deficient mice.

    PubMed

    Schmid, Rainer; Bosserhoff, Anja-Katrin

    2014-02-01

    Recent in vitro analysis of MIA/CD-RAP-deficient (MIA(-/-)) mesenchymal stem cells revealed altered chondrogenic differentiation, characterised by enhanced proliferation and delayed differentiation. However, adult MIA(-/-) mice develop normally and show only ultrastructural defects of the cartilage but no major abnormalities. We therefore focused, in this study, on chondrogenesis in vivo in MIA(-/-) mouse embryos to reveal potential molecular changes during embryogenesis and possible redundant mechanisms, which explain the almost normal phenotype despite MIA/CD-RAP loss. In situ hybridisation analysis revealed larger expression areas of Col2a1 and Sox9 positive, proliferating chondrocytes at day 15.5 and 16.5 of embryogenesis in MIA(-/-) mice. The initially diminished zone of Col10a1-expressing hypertrophic chondrocytes at day 15.5 was compensated at day 16.5 in MIA(-/-) embryos. Supported by in vitro studies using mesenchymal stem cells, we discovered that chondrogenesis in MIA(-/-) mice is modified by enhanced Sox9, Sox6 and AP-2α expression. Finally, we identified reduced AP1 and CRE activity, analysed by reporter gene- and electrophoretic mobility shift assays, important for redundancy mechanism which rescued delayed hypertrophic differentiation and allows normal development of MIA(-/-) mice. In summary, as observed in other knockout models of molecules important for cartilage development and differentiation, viability and functional integrity is reached by remarkable molecular redundancy in MIA/CD-RAP knockout mice.

  2. Partnering for environmental restoration: The Port Hope Harbour Remedial Action Plan (RAP)

    SciTech Connect

    Weston, S.M.C.

    1995-12-31

    A Remedial Action Plan (RAP) is being developed for Port Hope Harbour, one of 43 Areas of Concern (AOCs) identified by the International Joint Commission (IJC). The RAP, when implemented, will lead to the restoration and protection of desirable water conditions in Port Hope Harbour. The environmental concern associated with the harbor can be best viewed as a historical contaminated sediment problem. Approximately 90,000 m{sup 3} of sediment located in Port Hope Harbour`s turning basin and west slip are contaminated by uranium and thorium series radionuclides, heavy metals, and PCBs. There are several groups contributing to the development of the RAP. All of these groups have the common goal of developing an environmentally sound plan that reflects the views of the community. Strategic partnerships have been established that recognize the need to integrate and coordinate the efforts of all agencies, stakeholders, and the community. The objective is to develop an environmentally sound remediation plan through an efficient and effective management framework.

  3. Validation and relevance of Rheumatoid Arthritis Pain Scale (RAPS) in Indian (Asian) patients suffering from rheumatoid arthritis.

    PubMed

    Kianifard, Toktam; Kianyfard, Taghi; Chopra, Arvind

    2016-01-01

    Pain in RA is multifaceted and complex. Measuring instruments are inadequate. Rheumatoid Arthritis Pain Scale (RAPS) (Arthritis Care Res 45:317-323, 2001) was designed to measure pain comprehensively but has been sparsely reported. We decided to validate a suitable version for our community. Post translation (contextual), RAPS was administered (face to face interview) to 172 consenting patients of moderately severe RA (mean pain visual analogue scale (VAS) 5.4 cm) in a cross-sectional study using standard rheumatology case record form. RAPS contained 24 questions (numeric score, anchored at 0 (never) and 6 (always); range 0-144). Fifty-seven cohort patients on supervised rheumatology care were followed for 16 weeks. SPSS (v16) was used for statistical analysis, significant p < 0.05. RAPS showed good face and content validity (consensus). Construct/criterion validity was demonstrated for subclass domains and total RAPS (Cronbach's alpha 0.91, test-retest interclass correlation (Pearson) 0.71). Fair to modest correlation (p < 0.05) was seen with swollen joint count (0.16), Indian health assessment questionnaire (0.23), medical outcome short form (SF), 36 physical score (-0.35), SF 36 mental score (-0.21) and C-reactive protein (0.25), not with pain VAS. Similar results were shown for subclass domains (physiologic, affective, sensory, cognitive), except low alpha for affective. Age, disease duration and SF 36 were significant predictors (linear regression). In factor analysis, RAPS loaded with SF 36. The standardized response mean (0.6) was equal to pain VAS and DAS 28. RAPS was found to be a valid and clinically relevant instrument for measuring pain in Indian patients suffering from RA. It merits more widespread clinical use.

  4. Pharmacological Activation of Rap1 Antagonizes the Endothelial Barrier Disruption Induced by Exotoxins ExoS and ExoT of Pseudomonas aeruginosa

    PubMed Central

    Bouillot, Stéphanie; Attrée, Ina

    2015-01-01

    Most clinical strains of Pseudomonas aeruginosa, a leading agent of nosocomial infections, are multiresistant to antibiotherapy. Because of the paucity of new available antibiotics, the investigation of strategies aimed at limiting the action of its major virulence factors has gained much interest. The type 3 secretion system of P. aeruginosa and its effectors are known to be major determinants of toxicity and are required for bacterial dissemination in the host. Bacterial transmigration across the vascular wall is considered to be an important step in the infectious process. Using human endothelial primary cells, we demonstrate that forskolin (FSK), a drug inducing cyclic AMP (cAMP) elevation in eukaryotic cells, strikingly reduced the cell retraction provoked by two type 3 toxins, ExoS and ExoT, found in the majority of clinical strains. Conversely, cytotoxicity of a strain carrying the type 3 effector ExoU was unaffected by FSK. In addition, FSK altered the capacity of two ExoS/ExoT strains to transmigrate across cell monolayers. In agreement with these findings, other drugs and a cytokine inducing the increase of cAMP intracellular levels have also protected cells from retraction. cAMP is an activator of both protein kinase A and EPAC, a GTPase exchange factor of Rap1. Using activators or inhibitors of either pathway, we show that the beneficial effect of FSK is exerted by the activation of the EPAC/Rap1 axis, suggesting that its protective effect is mediated by reinforcing cell-cell and cell-substrate adhesion. PMID:25690098

  5. Drosophila-Cdh1 (Rap/Fzr) a regulatory subunit of APC/C is required for synaptic morphology, synaptic transmission and locomotion.

    PubMed

    Wise, Alexandria; Schatoff, Emma; Flores, Julian; Hua, Shao-Ying; Ueda, Atsushi; Wu, Chun-Fang; Venkatesh, Tadmiri

    2013-11-01

    The assembly of functional synapses requires the orchestration of the synthesis and degradation of a multitude of proteins. Protein degradation and modification by the conserved ubiquitination pathway has emerged as a key cellular regulatory mechanism during nervous system development and function (Kwabe and Brose, 2011). The anaphase promoting complex/cyclosome (APC/C) is a multi-subunit ubiquitin ligase complex primarily characterized for its role in the regulation of mitosis (Peters, 2002). In recent years, a role for APC/C in nervous system development and function has been rapidly emerging (Stegmuller and Bonni, 2005; Li et al., 2008). In the mammalian central nervous system the activator subunit, APC/C-Cdh1, has been shown to be a regulator of axon growth and dendrite morphogenesis (Konishi et al., 2004). In the Drosophila peripheral nervous system (PNS), APC2, a ligase subunit of the APC/C complex has been shown to regulate synaptic bouton size and activity (van Roessel et al., 2004). To investigate the role of APC/C-Cdh1 at the synapse we examined loss-of-function mutants of Rap/Fzr (Retina aberrant in pattern/Fizzy related), a Drosophila homolog of the mammalian Cdh1 during the development of the larval neuromuscular junction in Drosophila. Our cell biological, ultrastructural, electrophysiological, and behavioral data showed that rap/fzr loss-of-function mutations lead to changes in synaptic structure and function as well as locomotion defects. Data presented here show changes in size and morphology of synaptic boutons, and, muscle tissue organization. Electrophysiological experiments show that loss-of-function mutants exhibit increased frequency of spontaneous miniature synaptic potentials, indicating a higher rate of spontaneous synaptic vesicle fusion events. In addition, larval locomotion and peristaltic movement were also impaired. These findings suggest a role for Drosophila APC/C-Cdh1 mediated ubiquitination in regulating synaptic morphology

  6. Reactive atom plasma (RAP) processing of mirrors for astronomy

    NASA Astrophysics Data System (ADS)

    Subrahmanyan, Pradeep K.; Gardopée, George

    2008-07-01

    Modern day telescopes for astronomy have very complex requirements. Both ground and space based telescopes are getting much larger placing significant productivity requirements on the manufacturing processes employed. Conventional manufacturing paradigms involving mechanical abrasion have limitations related primarily to the material removal mechanisms employed. Reactive Atom Plasma (RAPTM) processing is a sub-aperture, non-contact, deterministic figuring technology performed at atmospheric pressures. The process has high material removal rates, and given the non-contact and atmospheric nature lends itself very well to scaling up for large aperture mirrors/segments. The process also benefits from its ability to simultaneously remove sub-surface damage (SSD) while imparting the desired figure to the surface. Developments are under way currently to scale the process up towards larger clear apertures while being able to figure in high spatial frequency features.

  7. Role of Epac2A/Rap1 signaling in interplay between incretin and sulfonylurea in insulin secretion.

    PubMed

    Takahashi, Harumi; Shibasaki, Tadao; Park, Jae-Hyung; Hidaka, Shihomi; Takahashi, Toshimasa; Ono, Aika; Song, Dae-Kyu; Seino, Susumu

    2015-04-01

    Incretin-related drugs and sulfonylureas are currently used worldwide for the treatment of type 2 diabetes. We recently found that Epac2A, a cAMP binding protein having guanine nucleotide exchange activity toward Rap, is a target of both incretin and sulfonylurea. This suggests the possibility of interplay between incretin and sulfonylurea through Epac2A/Rap1 signaling in insulin secretion. In this study, we examined the combinatorial effects of incretin and various sulfonylureas on insulin secretion and activation of Epac2A/Rap1 signaling. A strong augmentation of insulin secretion by combination of GLP-1 and glibenclamide or glimepiride, which was found in Epac2A(+/+) mice, was markedly reduced in Epac2A(-/-) mice. In contrast, the combinatorial effect of GLP-1 and gliclazide was rather mild, and the effect was not altered by Epac2A ablation. Activation of Rap1 was enhanced by the combination of an Epac-selective cAMP analog with glibenclamide or glimepiride but not gliclazide. In diet-induced obese mice, ablation of Epac2A reduced the insulin secretory response to coadministration of the GLP-1 receptor agonist liraglutide and glimepiride. These findings clarify the critical role of Epac2A/Rap1 signaling in the augmenting effect of incretin and sulfonylurea on insulin secretion and provide the basis for the effects of combination therapies of incretin-related drugs and sulfonylureas. PMID:25315008

  8. RAP, the sole octotricopeptide repeat protein in Arabidopsis, is required for chloroplast 16S rRNA maturation.

    PubMed

    Kleinknecht, Laura; Wang, Fei; Stübe, Roland; Philippar, Katrin; Nickelsen, Jörg; Bohne, Alexandra-Viola

    2014-02-01

    The biogenesis and activity of chloroplasts in both vascular plants and algae depends on an intracellular network of nucleus-encoded, trans-acting factors that control almost all aspects of organellar gene expression. Most of these regulatory factors belong to the helical repeat protein superfamily, which includes tetratricopeptide repeat, pentatricopeptide repeat, and the recently identified octotricopeptide repeat (OPR) proteins. Whereas green algae express many different OPR proteins, only a single orthologous OPR protein is encoded in the genomes of most land plants. Here, we report the characterization of the only OPR protein in Arabidopsis thaliana, RAP, which has previously been implicated in plant pathogen defense. Loss of RAP led to a severe defect in processing of chloroplast 16S rRNA resulting in impaired chloroplast translation and photosynthesis. In vitro RNA binding and RNase protection assays revealed that RAP has an intrinsic and specific RNA binding capacity, and the RAP binding site was mapped to the 5' region of the 16S rRNA precursor. Nucleoid localization of RAP was shown by transient green fluorescent protein import assays, implicating the nucleoid as the site of chloroplast rRNA processing. Taken together, our data indicate that the single OPR protein in Arabidopsis is important for a basic process of chloroplast biogenesis.

  9. Installation and operation of a large scale RAPS system in Peru

    NASA Astrophysics Data System (ADS)

    Cole, J. F.

    In 1997, International Lead Zinc Research Organization Inc. (ILZRO), Solar Energy Industries Association (SEIA), and the Ministry of Energy and Mines (MEM) of Peru signed a Memorandum of Understanding to facilitate the installation of hybrid remote area power supply (RAPS) systems in the Amazon region of Peru. Many remote villages in this vast region have either no or limited electricity supplied by diesel generators running a few hours per day. Subsequently, ILZRO sponsored the engineering design of the hybrid RAPS system and SEIA supported a socio-economic study to determine the sustainability of such systems and the locations for pilot installations. In mid-1998, the Peruvian government approved the design of the system. ILZRO then began efforts to obtain governmental and inter-governmental funding to supplement its own funds to underwrite the cost of manufacture and installation of the systems in two villages in the Amazon region. Additional major funding has been received from the Global Environmental Facility (GEF) administered by the United Nations Development Program (UNDP) and from the Common Fund for Commodities (CFC). Funds have also been received from the US Department of Energy, the International Greenhouse Partnership (Australia) and the Peruvian government. The RAPS system consists of modules designed to provide 150 kW h per day of utility grade ac electricity over a 24 h period. Each module contains a diesel generator, battery bank using heavy-duty 2 V VRLA GEL batteries, a battery charger, a photovoltaic array and an ac/dc inverter. The batteries and electrical components are housed in modified shipping containers. The modules can be installed with a new generator or retrofitted to an existing generator. The charging and discharging regime of the batteries has been recommended by a study carried out by CSIRO, which has simulated the RAPS operation. The system will employ a partial-state-of-charge (PSOC) regime in order to optimize the life of the

  10. Cep169, a Novel Microtubule Plus-End-Tracking Centrosomal Protein, Binds to CDK5RAP2 and Regulates Microtubule Stability

    PubMed Central

    Mori, Yusuke; Inoue, Yoko; Tanaka, Sayori; Doda, Satoka; Yamanaka, Shota; Fukuchi, Hiroki; Terada, Yasuhiko

    2015-01-01

    The centrosomal protein, CDK5RAP2, is a microcephaly protein that regulates centrosomal maturation by recruitment of a γ-tubulin ring complex (γ-TuRC) onto centrosomes. In this report, we identified a novel human centrosomal protein, Cep169, as a binding partner of CDK5RAP2, a member of microtubule plus-end-tracking proteins (+TIPs). Cep169 interacts directly with CDK5RAP2 through CM1, an evolutionarily conserved domain, and colocalizes at the pericentriolar matrix (PCM) around centrioles with CDK5RAP2. In addition, Cep169 interacts with EB1 through SxIP-motif responsible for EB1 binding, and colocalizes with CDK5RAP2 at the microtubule plus-end. EB1-binding–deficient Cep169 abolishes EB1 interaction and microtubule plus-end attachment, indicating Cep169 as a novel member of +TIPs. We further show that ectopic expression of either Cep169 or CDK5RAP2 induces microtubule bundling and acetylation in U2OS cells, and depletion of Cep169 induces microtubule depolymerization in HeLa cells, although Cep169 is not required for assembly of γ-tubulin onto centrosome by CDK5RAP2. These results show that Cep169 targets microtubule tips and regulates stability of microtubules with CDK5RAP2. PMID:26485573

  11. Cep169, a Novel Microtubule Plus-End-Tracking Centrosomal Protein, Binds to CDK5RAP2 and Regulates Microtubule Stability.

    PubMed

    Mori, Yusuke; Inoue, Yoko; Tanaka, Sayori; Doda, Satoka; Yamanaka, Shota; Fukuchi, Hiroki; Terada, Yasuhiko

    2015-01-01

    The centrosomal protein, CDK5RAP2, is a microcephaly protein that regulates centrosomal maturation by recruitment of a γ-tubulin ring complex (γ-TuRC) onto centrosomes. In this report, we identified a novel human centrosomal protein, Cep169, as a binding partner of CDK5RAP2, a member of microtubule plus-end-tracking proteins (+TIPs). Cep169 interacts directly with CDK5RAP2 through CM1, an evolutionarily conserved domain, and colocalizes at the pericentriolar matrix (PCM) around centrioles with CDK5RAP2. In addition, Cep169 interacts with EB1 through SxIP-motif responsible for EB1 binding, and colocalizes with CDK5RAP2 at the microtubule plus-end. EB1-binding-deficient Cep169 abolishes EB1 interaction and microtubule plus-end attachment, indicating Cep169 as a novel member of +TIPs. We further show that ectopic expression of either Cep169 or CDK5RAP2 induces microtubule bundling and acetylation in U2OS cells, and depletion of Cep169 induces microtubule depolymerization in HeLa cells, although Cep169 is not required for assembly of γ-tubulin onto centrosome by CDK5RAP2. These results show that Cep169 targets microtubule tips and regulates stability of microtubules with CDK5RAP2. PMID:26485573

  12. Induction of Plasmid Conjugation in Bacillus subtilis Is Bistable and Driven by a Direct Interaction of a Rap/Phr Quorum-sensing System with a Master Repressor.

    PubMed

    Rösch, Thomas C; Graumann, Peter L

    2015-08-14

    Conjugation of plasmid pLS20 from Bacillus subtilis is limited to a time window between early and late exponential growth. Genetic evidence has suggested that pLS20-encoded protein RcoLS20 represses expression of a large conjugation operon, whereas Rap protein RapLS20 relieves repression. We show that RapLS20 is a true antirepressor protein that forms dimers in vivo and in vitro and that it directly binds to the repressor protein RcoLS20 in a 1:1 stoichiometry. We provide evidence that RapLS20 binds to the helix-turn-helix-containing domain of RcoLS20 in vivo, probably obstructing DNA binding of RcoLS20, as seen in competitive DNA binding experiments. The activity of RapLS20 in turn is counteracted by the addition of the cognate PhrLS20 peptide, which directly binds to the Rap protein and presumably induces a conformational change of the antirepressor. Thus, a Rap protein acts directly as an antirepressor protein during regulation of plasmid conjugation, turning on conjugation, and is counteracted by the PhrLS20 peptide, which, by analogy to known Rap/Phr systems, is secreted and taken back up into the cells, mediating cell density-driven regulation. Finally, we show that this switchlike process establishes a population heterogeneity, where up to 30% of the cells induce transcription of the conjugation operon.

  13. An Action Research Study on the Influence of Gangsta Rap on Academic and Behavioral Issues of 5th Grade African-American Males

    ERIC Educational Resources Information Center

    Lewis, Shaun; Boes, Susan R.; Chibbaro, Julie S.

    2015-01-01

    This small action research study (ARS) began with a review of the literature examining the relationship of gangsta rap in regards to academic achievement, self-esteem, decision-making, identity issues and development of young African American males. The purpose of the ARS was to examine the correlation between gangsta rap and its influence on 5th…

  14. Geeksta Rap: An Example of the Utilization of Elements of Popular Culture in Science Education and Outreach

    NASA Astrophysics Data System (ADS)

    Hall, F.; Otto, A.

    2003-12-01

    Science education and outreach efforts must compete for the attention of students in an environment filled with many distractions. Many of the most compelling of these distractions arise from popular culture. Rather than bemoaning this situation, we propose that we make use of some of those elements of popular culture which garner the most attention. In particular, we propose that we utilize the popularity of current hip-hop and rap music in science education and outreach efforts. To that end, we present some examples of what we call `geeksta rap' and discuss some of the considerations relevant for its preparation. We also discuss various uses of `geeksta rap' in science education and outreach.

  15. Wet electrostatic precipitator having removable nested hexagonal collector plates and magnetic aligning and rapping means

    SciTech Connect

    Young, C.E.; Drzewiecki, G.

    1984-04-10

    A wet electrostatic precipitator including a plurality of removable nested collecting electrodes or plates forming a repeating pattern of hexagonal collecting zones throughout the precipitator. Each collecting plate is formed with a sixty degree bend along two opposing longitudinal edges so as to allow three plates to form a self-nesting Y-shaped intersection point. Six points form a hexagonal collecting zone. The plates are removable thereby expediting replacement. A plurality of strategically placed spray nozzles provide wash fluid to the plates. Magnet sets provide for discharge electrode alignment and rapping.

  16. A Software Demonstration of 'rap': Preparing CAD Geometries for Overlapping Grid Generation

    SciTech Connect

    Anders Petersson, N.

    2002-02-15

    We demonstrate the application code ''rap'' which is part of the ''Overture'' library. A CAD geometry imported from an IGES file is first cleaned up and simplified to suit the needs of mesh generation. Thereafter, the topology of the model is computed and a water-tight surface triangulation is created on the CAD surface. This triangulation is used to speed up the projection of points onto the CAD surface during the generation of overlapping surface grids. From each surface grid, volume grids are grown into the domain using a hyperbolic marching procedure. The final step is to fill any remaining parts of the interior with background meshes.

  17. Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization

    PubMed Central

    Wang, Haibo; Han, Xiaokun; Bretz, Colin A; Becker, Silke; Gambhir, Deeksha; Smith, George W; Samulski, R Jude; Wittchen, Erika S; Quilliam, Lawrence A; Chrzanowska-Wodnicka, Magdalena; Hartnett, M Elizabeth

    2016-01-01

    To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 108 viral particle/µl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes. PMID:27606349

  18. Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization.

    PubMed

    Wang, Haibo; Han, Xiaokun; Bretz, Colin A; Becker, Silke; Gambhir, Deeksha; Smith, George W; Samulski, R Jude; Wittchen, Erika S; Quilliam, Lawrence A; Chrzanowska-Wodnicka, Magdalena; Hartnett, M Elizabeth

    2016-01-01

    To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 10(8) viral particle/µl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes.

  19. Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization

    PubMed Central

    Wang, Haibo; Han, Xiaokun; Bretz, Colin A; Becker, Silke; Gambhir, Deeksha; Smith, George W; Samulski, R Jude; Wittchen, Erika S; Quilliam, Lawrence A; Chrzanowska-Wodnicka, Magdalena; Hartnett, M Elizabeth

    2016-01-01

    To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 108 viral particle/µl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes.

  20. Interactions of the HIV-1 Tat and RAP74 proteins with the RNA polymerase II CTD phosphatase FCP1.

    PubMed

    Abbott, Karen L; Archambault, Jacques; Xiao, Hua; Nguyen, Bao D; Roeder, Robert G; Greenblatt, Jack; Omichinski, James G; Legault, Pascale

    2005-03-01

    FCP1, a phosphatase specific for the carboxyl-terminal domain of the largest subunit of RNA polymerase II, is regulated by the HIV-1 Tat protein, CK2, TFIIB, and the large subunit of TFIIF (RAP74). We have characterized the interactions of Tat and RAP74 with the BRCT-containing central domain of FCP1 (FCP1(562)(-)(738)). We demonstrated that FCP1 is required for Tat-mediated transactivation in vitro and that amino acids 562-685 of FCP1 are necessary for Tat interaction in yeast two-hybrid studies. From sequence alignments, we identified a conserved acidic/hydrophobic region in FCP1 adjacent to its highly conserved BRCT domain. In vitro binding studies with purified proteins indicate that HIV-1 Tat interacts with both the acidic/hydrophobic region and the BRCT domain of FCP1, whereas RAP74(436)(-)(517) interacts solely with a portion of the acidic/hydrophobic region containing a conserved LXXLL-like motif. HIV-1 Tat inhibits the binding of RAP74(436)(-)(517) to FCP1. In a companion paper (K. Abbott et al. (2005) Enhanced Binding of RNAPII CTD Phosphatase FCP1 to RAP74 Following CK2 Phosphorylation, Biochemistry 44, 2732-2745, we identified a novel CK2 site adjacent to this conserved LXXLL-like motif. Phosphorylation of FCP1(562)(-)(619) by CK2 at this site increases binding to RAP74(436)(-)(517), but this phosphorylation is inhibited by Tat. Our results provide insights into the mechanisms by which Tat inhibits the FCP1 CTD phosphatase activity and by which FCP1 mediates transcriptional activation by Tat. In addition to increasing our understanding of the role of HIV-1 Tat in transcriptional regulation, this study defines a clear role for regions adjacent to the BRCT domain in promoting important protein-protein interactions. PMID:15723517

  1. Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization.

    PubMed

    Wang, Haibo; Han, Xiaokun; Bretz, Colin A; Becker, Silke; Gambhir, Deeksha; Smith, George W; Samulski, R Jude; Wittchen, Erika S; Quilliam, Lawrence A; Chrzanowska-Wodnicka, Magdalena; Hartnett, M Elizabeth

    2016-01-01

    To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 10(8) viral particle/µl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes. PMID:27606349

  2. Synaptotagmin-like protein 1 interacts with the GTPase-activating protein Rap1GAP2 and regulates dense granule secretion in platelets.

    PubMed

    Neumüller, Olga; Hoffmeister, Meike; Babica, Jan; Prelle, Carola; Gegenbauer, Kristina; Smolenski, Albert P

    2009-08-13

    The small guanine-nucleotide-binding protein Rap1 plays a key role in platelet aggregation and hemostasis, and we recently identified Rap1GAP2 as the only GTPase-activating protein of Rap1 in platelets. In search of Rap1GAP2-associated proteins, we performed yeast-2-hybrid screening and found synaptotagmin-like protein 1 (Slp1) as a new binding partner. We confirmed the interaction of Rap1GAP2 and Slp1 in transfected COS-1 and HeLa cells and at endogenous level in human platelets. Mapping studies showed that Rap1GAP2 binds through amino acids T524-K525-X-T527 within its C-terminus to the C2A domain of Slp1. Slp1 contains a Rab27-binding domain, and we demonstrate that Rap1GAP2, Slp1, and Rab27 form a trimeric complex in transfected cells and in platelets. Purified Slp1 dose-dependently decreased dense granule secretion in streptolysin-O-permeabilized platelets stimulated with calcium or guanosine 5'-O-[gamma-thio] triphosphate. The isolated C2A domain of Slp1 had a stimulatory effect on granule secretion and reversed the inhibitory effect of full-length Slp1. Purified Rap1GAP2 augmented dense granule secretion of permeabilized platelets, whereas deletion of the Slp1-binding TKXT motif abolished the effect of Rap1GAP2. We conclude that Slp1 inhibits dense granule secretion in platelets and that Rap1GAP2 modulates secretion by binding to Slp1. PMID:19528539

  3. RapA2 Is a Calcium-binding Lectin Composed of Two Highly Conserved Cadherin-like Domains That Specifically Recognize Rhizobium leguminosarum Acidic Exopolysaccharides*

    PubMed Central

    Abdian, Patricia L.; Caramelo, Julio J.; Ausmees, Nora; Zorreguieta, Angeles

    2013-01-01

    In silico analyses have revealed a conserved protein domain (CHDL) widely present in bacteria that has significant structural similarity to eukaryotic cadherins. A CHDL domain was shown to be present in RapA, a protein that is involved in autoaggregation of Rhizobium cells, biofilm formation, and adhesion to plant roots as shown by us and others. Structural similarity to cadherins suggested calcium-dependent oligomerization of CHDL domains as a mechanistic basis for RapA action. Here we show by circular dichroism spectroscopy, light scattering, isothermal titration calorimetry, and other methods that RapA2 from Rhizobium leguminosarum indeed exhibits a cadherin-like β-sheet conformation and that its proper folding and stability are dependent on the binding of one calcium ion per protein molecule. By further in silico analysis we also reveal that RapA2 consists of two CHDL domains and expand the range of CHDL-containing proteins in bacteria and archaea. However, light scattering assays at various concentrations of added calcium revealed that RapA2 formed neither homo-oligomers nor hetero-oligomers with RapB (a distinct CHDL protein), indicating that RapA2 does not mediate cellular interactions through a cadherin-like mechanism. Instead, we demonstrate that RapA2 interacts specifically with the acidic exopolysaccharides (EPSs) produced by R. leguminosarum in a calcium-dependent manner, sustaining a role of these proteins in the development of the biofilm matrix made of EPS. Because EPS binding by RapA2 can only be attributed to its two CHDL domains, we propose that RapA2 is a calcium-dependent lectin and that CHDL domains in various bacterial and archaeal proteins confer carbohydrate binding activity to these proteins. PMID:23235153

  4. Linkage-specific avidity defines the lysine 63-linked polyubiquitin binding preference of Rap80

    PubMed Central

    Sims, Joshua J.; Cohen, Robert E.

    2009-01-01

    Summary Linkage-specific polyubiquitin recognition is thought to make possible the diverse set of functional outcomes associated with ubiquitination. Thus far, mechanistic insight into this selectivity has been largely limited to single domains that preferentially bind to lysine 48-linked polyubiquitin (K48-polyUb) in isolation. Here we propose a mechanism, linkage-specific avidity, in which multiple ubiquitin binding domains are arranged in space so that simultaneous, high-affinity interactions are optimum with one polyUb linkage, but unfavorable or impossible with other polyUb topologies and monoUb. Our model is human Rap80, which contains tandem ubiquitin interacting motifs (UIMs) that bind to K63-polyUb at DNA double-strand breaks. We show how the sequence between the Rap80 UIMs positions the domains for efficient, avid binding across a single K63 linkage, thus defining selectivity. We also demonstrate K48-specific avidity in a different protein, ataxin-3. Using tandem UIMs, we establish the general principles governing polyUb linkage selectivity and affinity in multivalent ubiquitin receptors. PMID:19328070

  5. Clinical evaluation of the RapID-ANA II panel for identification of anaerobic bacteria.

    PubMed

    Celig, D M; Schreckenberger, P C

    1991-03-01

    The accuracy of the RapID-ANA II system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) was evaluated by comparing the results obtained with that system with results obtained by the methods described by the Virginia Polytechnic Institute and State University. Three hundred anaerobic bacteria were tested, including 259 clinical isolates and 41 stock strains of anaerobic microorganisms representing 16 genera and 48 species. When identifications to the genus level only were included, 96% of the anaerobic gram-negative bacilli, 94% of the Clostridium species, 83% of the anaerobic, nonsporeforming, gram-positive bacilli, and 97% of the anaerobic cocci were correctly identified. When correct identifications to the genus and species levels were compared, 86% of 152 anaerobic gram-negative bacilli, 76% of 34 Clostridium species, 81% of 41 anaerobic, nonsporeforming, gram-positive bacilli, and 97% of 73 anaerobic cocci were correctly identified. Eight isolates (3%) produced inadequate identification in which the correct identification was listed with one or two other possible choices and extra tests were required for separation. A total of 9 isolates (3%) were misidentified by the RapID-ANA II panel. Overall, the system was able to correctly identify 94% of all the isolates to the genus level and 87% of the isolates to the species level in 4 h by using aerobic incubation.

  6. Ensemble covariances adaptively localized with ECO-RAP. Part 1: tests on simple error models

    NASA Astrophysics Data System (ADS)

    Bishop, Craig H.; Hodyss, Daniel

    2009-01-01

    In atmospheric data assimilation (DA), observations over a 6-12-h time window are used to estimate the state. Non-adaptive moderation or localization functions are widely used in ensemble DA to reduce the amplitude of spurious ensemble correlations. These functions are inappropriate (1) if true error correlation functions move a comparable distance to the localization length scale over the time window and/or (2) if the widths of true error correlation functions are highly flow dependent. A method for generating localization functions that move with the true error correlation functions and that also adapt to the width of the true error correlation function is given. The method uses ensemble correlations raised to a power (ECO-RAP). A gallery of periodic one-dimensional error models is used to show how the method uses error propagation information and error correlation width information retained by powers of raw ensemble correlations to propagate and adaptively adjust the width of the localization function. It is found that ECO-RAP localization outperforms non-adaptive localization when the true errors are propagating or the error correlation length scale is varying and is as good as non-adaptive localization when such variations in error covariance structure are absent.

  7. Rap as a roadway: creating creolized forms of science in an era of cultural globalization

    NASA Astrophysics Data System (ADS)

    Elmesky, Rowhea

    2011-03-01

    Even during an era of cultural globalization where diversity, hybridity, and heterogeneity prevail, educational institutions remain unchanged and economically and racially marginalized students continue to experience a sense of exclusion in school. Whereas the science education community often addresses such exclusion in terms of the achievement gap or the lack of materials and qualified teachers in urban schools, there are also more subtle ways in which these students remain as outsiders to the culture of science. The study highlights how the acceptance and affordance of students' cultural capital can encourage a sense of belonging with school science. Specifically, this paper contributes to the literature by sharing longitudinal findings that reveal students' skills of orality, in the form of rap practices, can be rich resources for developing creolized forms of school science, and how rap creates entryways for students to form and reform hybridized identities in which canonical science discourse and lyrics about non-science subjects can begin to emerge in integrated, fluid and seamless manners.

  8. Catheter Ablation of Right-Sided Accessory Pathways in Adults Using the Three-Dimensional Mapping System: A Randomized Comparison to the Conventional Approach.

    PubMed

    Ma, Yuedong; Qiu, Jia; Yang, Yang; Tang, Anli

    2015-01-01

    Three-dimensional (3D) mapping and navigation systems have been widely used for the ablation of atrial fibrillation and ventricular tachycardia, but the applicability of these systems for the ablation of supraventricular tachycardia (SVT) due to right-sided accessory pathways (RAPs) remains unknown. The goal of this prospective randomized study was to compare the safety, efficiency, and efficacy of nonfluoroscopic and conventional fluoroscopic mapping techniques in guiding catheter ablation of SVT due to RAPs. Of the 393 consecutive patients with SVT who were randomized to receive either conventional fluoroscopic or Ensite NavX mapping-guided ablation, 64 patients with RAPs were included for analysis. Endpoints for ablation were no evidence of RAP conduction and no inducible atrioventricular reentrant tachycardia (AVRT). The 3D group showed fewer ablation pulses and a shorter total ablation time compared to the conventional group, although the acute procedural success did not differ significantly between the two groups. Total procedure time, electrophysiological study time, total fluoroscopy time, and cumulative radiation doses were also significantly reduced in the 3D group. Patients in the conventional group with a right atrium diameter (RAD) ≥ 47 mm required a longer fluoroscopy time. There was no significant difference in the recurrence rates between the two groups over a follow-up period of 3 to 29 months. There were no permanent complications. The 3D mapping system may be a preferred alternative for patients with AVRT due to RAPs, especially for patients with a large RAD (≥ 47 mm).

  9. Catheter Ablation of Right-Sided Accessory Pathways in Adults Using the Three-Dimensional Mapping System: A Randomized Comparison to the Conventional Approach

    PubMed Central

    Tang, Anli

    2015-01-01

    Three-dimensional (3D) mapping and navigation systems have been widely used for the ablation of atrial fibrillation and ventricular tachycardia, but the applicability of these systems for the ablation of supraventricular tachycardia (SVT) due to right-sided accessory pathways (RAPs) remains unknown. The goal of this prospective randomized study was to compare the safety, efficiency, and efficacy of nonfluoroscopic and conventional fluoroscopic mapping techniques in guiding catheter ablation of SVT due to RAPs. Of the 393 consecutive patients with SVT who were randomized to receive either conventional fluoroscopic or Ensite NavX mapping-guided ablation, 64 patients with RAPs were included for analysis. Endpoints for ablation were no evidence of RAP conduction and no inducible atrioventricular reentrant tachycardia (AVRT). The 3D group showed fewer ablation pulses and a shorter total ablation time compared to the conventional group, although the acute procedural success did not differ significantly between the two groups. Total procedure time, electrophysiological study time, total fluoroscopy time, and cumulative radiation doses were also significantly reduced in the 3D group. Patients in the conventional group with a right atrium diameter (RAD) ≥ 47 mm required a longer fluoroscopy time. There was no significant difference in the recurrence rates between the two groups over a follow-up period of 3 to 29 months. There were no permanent complications. The 3D mapping system may be a preferred alternative for patients with AVRT due to RAPs, especially for patients with a large RAD (≥ 47 mm). PMID:26083408

  10. AgrAbility Project

    MedlinePlus

    ... It’s About Hope AgrAbility on Twitter AgrAbility on Facebook AgrAbility on You Tube AgrAbility… It’s About Hope ... anniversary throughout 2016... AgrAbility Harvest Get a copy Facebook Posts National AgrAbility Project 5 days ago AgrAbility's ...

  11. Open-Source Wax RepRap 3-D Printer for Rapid Prototyping Paper-Based Microfluidics.

    PubMed

    Pearce, J M; Anzalone, N C; Heldt, C L

    2016-08-01

    The open-source release of self-replicating rapid prototypers (RepRaps) has created a rich opportunity for low-cost distributed digital fabrication of complex 3-D objects such as scientific equipment. For example, 3-D printable reactionware devices offer the opportunity to combine open hardware microfluidic handling with lab-on-a-chip reactionware to radically reduce costs and increase the number and complexity of microfluidic applications. To further drive down the cost while improving the performance of lab-on-a-chip paper-based microfluidic prototyping, this study reports on the development of a RepRap upgrade capable of converting a Prusa Mendel RepRap into a wax 3-D printer for paper-based microfluidic applications. An open-source hardware approach is used to demonstrate a 3-D printable upgrade for the 3-D printer, which combines a heated syringe pump with the RepRap/Arduino 3-D control. The bill of materials, designs, basic assembly, and use instructions are provided, along with a completely free and open-source software tool chain. The open-source hardware device described here accelerates the potential of the nascent field of electrochemical detection combined with paper-based microfluidics by dropping the marginal cost of prototyping to nearly zero while accelerating the turnover between paper-based microfluidic designs. PMID:26763294

  12. 40 CFR 270.175 - For what reasons may the Director choose to modify my final RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... section exist(s). If one or more of these reasons do not exist, then the Director will not modify your... reasons to modify your compliance schedule, such as an act of God, strike, flood, or materials shortage or... exists that was unknown when the RAP was issued....

  13. Loss of CDK5RAP2 affects neural but not non-neural mESC differentiation into cardiomyocytes.

    PubMed

    Kraemer, Nadine; Ravindran, Ethiraj; Zaqout, Sami; Neubert, Gerda; Schindler, Detlev; Ninnemann, Olaf; Gräf, Ralph; Seiler, Andrea E M; Kaindl, Angela M

    2015-01-01

    Biallelic mutations in the gene encoding centrosomal CDK5RAP2 lead to autosomal recessive primary microcephaly (MCPH), a disorder characterized by pronounced reduction in volume of otherwise architectonical normal brains and intellectual deficit. The current model for the microcephaly phenotype in MCPH invokes a premature shift from symmetric to asymmetric neural progenitor-cell divisions with a subsequent depletion of the progenitor pool. The isolated neural phenotype, despite the ubiquitous expression of CDK5RAP2, and reports of progressive microcephaly in individual MCPH cases prompted us to investigate neural and non-neural differentiation of Cdk5rap2-depleted and control murine embryonic stem cells (mESC). We demonstrate an accumulating proliferation defect of neurally differentiating Cdk5rap2-depleted mESC and cell death of proliferative and early postmitotic cells. A similar effect does not occur in non-neural differentiation into beating cardiomyocytes, which is in line with the lack of non-central nervous system features in MCPH patients. Our data suggest that MCPH is not only caused by premature differentiation of progenitors, but also by reduced propagation and survival of neural progenitors. PMID:25942099

  14. CDK5RAP2 expression during murine and human brain development correlates with pathology in primary autosomal recessive microcephaly.

    PubMed

    Issa, Lina; Kraemer, Nadine; Rickert, Christian H; Sifringer, Marco; Ninnemann, Olaf; Stoltenburg-Didinger, Gisela; Kaindl, Angela M

    2013-09-01

    Homozygous mutations in the cyclin-dependent kinase-5 regulatory subunit-associated protein 2 gene CDK5RAP2 cause primary autosomal recessive microcephaly (MCPH). MCPH is characterized by a pronounced reduction of brain volume, particularly of the cerebral cortex, and mental retardation. Though it is a rare developmental disorder, MCPH has moved into the spotlight of neuroscience because of its proposed central role in stem-cell biology and brain development. Investigation of the neural basis of genetically defined MCPH has been limited to animal studies and neuroimaging of affected patients as no neuropathological studies have been published. In the present study, we depict the spatiotemporal expression of CDK5RAP2 in the developing brain of mouse and human. We found intriguing concordance between regions of high CDK5RAP2 expression in the mouse and sites of pathology suggested by neuroimaging studies in humans and mouse. Our findings in human tissue confirm those in mouse tissues, underlining the function of CDK5RAP2 in cell proliferation and arguing for a conserved role of this protein in the development of the mammalian cerebral cortex. PMID:22806269

  15. Cdk5rap2 interacts with pericentrin to maintain the neural progenitor pool in the developing neocortex.

    PubMed

    Buchman, Joshua J; Tseng, Huan-Chung; Zhou, Ying; Frank, Christopher L; Xie, Zhigang; Tsai, Li-Huei

    2010-05-13

    Primary autosomal-recessive microcephaly (MCPH) and Majewski osteodysplastic primordial dwarfism type II (MOPDII) are both genetic diseases that result in decreased brain size at birth. MCPH is thought to arise from alterations in the size of the neural progenitor pool, but the cause of this defect has not been thoroughly explored. We find that one of the genes associated with MCPH, Cdk5rap2, is highly expressed in the neural progenitor pool and that its loss results in a depletion of apical progenitors and increased cell-cycle exit leading to premature neuronal differentiation. We link Cdk5rap2 function to the pericentriolar material protein pericentrin, loss of function of which is associated with MOPDII. Depletion of pericentrin in neural progenitors phenocopies effects of Cdk5rap2 knockdown and results in decreased recruitment of Cdk5rap2 to the centrosome. Our findings uncover a common mechanism, involving aberrations in the neurogenesis program, that may underlie the development of microcephaly in multiple diseases. PMID:20471352

  16. A novel nonsense CDK5RAP2 mutation in a Somali child with primary microcephaly and sensorineural hearing loss.

    PubMed

    Pagnamenta, Alistair T; Murray, Jennie E; Yoon, Grace; Sadighi Akha, Elham; Harrison, Victoria; Bicknell, Louise S; Ajilogba, Kaseem; Stewart, Helen; Kini, Usha; Taylor, Jenny C; Keays, David A; Jackson, Andrew P; Knight, Samantha J L

    2012-10-01

    Primary microcephaly is a genetically heterogeneous condition characterized by reduced head circumference (-3 SDS or more) and mild-to-moderate learning disability. Here, we describe clinical and molecular investigations of a microcephalic child with sensorineural hearing loss. Although consanguinity was unreported initially, detection of 13.7 Mb of copy neutral loss of heterozygosity (cnLOH) on chromosome 9 implicated the CDK5RAP2 gene. Targeted sequencing identified a homozygous E234X mutation, only the third mutation to be described in CDK5RAP2, the first in an individual of non-Pakistani descent. Sensorineural hearing loss is not generally considered to be consistent with autosomal recessive microcephaly and therefore it seems likely that the deafness in this individual is caused by the co-occurrence of a further gene mutation, independent of CDK5RAP2. Nevertheless, further detailed clinical descriptions of rare CDK5RAP2 patients, including hearing assessments will be needed to resolve fully the phenotypic range associated with mutations in this gene. This study also highlights the utility of SNP-array testing to guide disease gene identification where an autosomal recessive condition is plausible. PMID:22887808

  17. Role of Epac1, an Exchange Factor for Rap GTPases, in Endothelial Microtubule Dynamics and Barrier Function

    PubMed Central

    Sehrawat, Seema; Cullere, Xavier; Patel, Sunita; Italiano, Joseph

    2008-01-01

    Rap1 GTPase activation by its cAMP responsive nucleotide exchange factor Epac present in endothelial cells increases endothelial cell barrier function with an associated increase in cortical actin. Here, Epac1 was shown to be responsible for these actin changes and to colocalize with microtubules in human umbilical vein endothelial cells. Importantly, Epac activation with a cAMP analogue, 8-pCPT-2′O-Me-cAMP resulted in a net increase in the length of microtubules. This did not require cell–cell interactions or Rap GTPase activation, and it was attributed to microtubule growth as assessed by time-lapse microscopy of human umbilical vein endothelial cell expressing fluorophore-linked microtubule plus-end marker end-binding protein 3. An intact microtubule network was required for Epac-mediated changes in cortical actin and barrier enhancement, but it was not required for Rap activation. Finally, Epac activation reversed microtubule-dependent increases in vascular permeability induced by tumor necrosis factor-α and transforming growth factor-β. Thus, Epac can directly promote microtubule growth in endothelial cells. This, together with Rap activation leads to an increase in cortical actin, which has functional significance for vascular permeability. PMID:18172027

  18. 40 CFR 270.125 - If I submit my RAP application as part of another document, what must I do?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... of another document, what must I do? 270.125 Section 270.125 Protection of Environment ENVIRONMENTAL... as part of another document, what must I do? If you submit your application for a RAP as a part of another document, you must clearly identify the components of that document that constitute your...

  19. 40 CFR 270.125 - If I submit my RAP application as part of another document, what must I do?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... of another document, what must I do? 270.125 Section 270.125 Protection of Environment ENVIRONMENTAL... as part of another document, what must I do? If you submit your application for a RAP as a part of another document, you must clearly identify the components of that document that constitute your...

  20. Utility of fecal calprotectin in differentiating inflammatory bowel disease (IBD) from recurrent abdominal pain (RAP) in children

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: It often is difficult to differentiate IBD from RAP in children. Fecal calprotectin concentration has been proposed as a marker to identify gastrointestinal inflammation and it may be useful in distinguishing organic disease (i.e., IBD) from normals. However, there are scant data regardi...

  1. Differential Gender Effects of Exposure to Rap Music on African American Adolescents' Acceptance of Teen Dating Violence.

    ERIC Educational Resources Information Center

    Johnson, James D.; And Others

    1995-01-01

    Assessed the effects of exposure to nonviolent rap videos on black adolescents' perceptions of teen dating violence. Results from 60 black adolescents and teenagers indicate a significant interaction between gender and video exposure: male acceptance of the use of violence was not a function of viewing the videos, whereas video-viewing females…

  2. Slp2-a controls renal epithelial cell size through regulation of Rap-ezrin signaling independently of Rab27.

    PubMed

    Yasuda, Takao; Fukuda, Mitsunori

    2014-02-01

    Synaptotagmin-like protein 2 (Slp2-a/Sytl2) is a Rab27 effector protein that regulates transport of Rab27-bearing vesicles and organelles through its N-terminal Rab27-binding domain and a phospholipid-binding C2A domain. Here we demonstrate a Rab27-independent function of Slp2-a in the control of renal cell size through a previously uncharacterized C2B domain. We found that by recruiting Rap1 GAPs to the plasma membrane of MDCK II cells through the C2B domain, Slp2-a inactivates Rap signaling and modulates the size of the cells. Functional ablation of Slp2-a resulted in an increase in the size of MDCK II cells. Drosophila Slp Bitesize was found to compensate for the function of Slp2-a in MDCK II cells, thereby indicating that the mechanism of the cell size control by Slp proteins has been evolutionarily conserved. Interestingly, blockade of the activity of ezrin, a downstream target of Rap, with the glucosylceramide synthase inhibitor, miglustat, effectively inhibited cell spreading of Slp2-a-knockdown cells. We also discovered aberrant expression of Slp2-a and increased activity of ezrin in pcy (Nphp3(pcy)) mice, a model of polycystic kidney disease that is characterized by renal cell spreading. Our findings indicate that Slp2-a controls renal cell size through regulation of Rap-ezrin signaling independently of Rab27.

  3. The Effect of Using Rapping To Teach Selected Musical Forms to Urban African American Middle School Students.

    ERIC Educational Resources Information Center

    Akintunde, Omowale

    A study determined the effects of a pedagogical approach using rap music on the learning of musical forms among urban African American youth and whether there were differential effects among students of different levels of self-esteem. Urban African American youth (n=66) from the St. Louis County Public Schools who were enrolled in general music…

  4. Loss of CDK5RAP2 affects neural but not non-neural mESC differentiation into cardiomyocytes

    PubMed Central

    Kraemer, Nadine; Ravindran, Ethiraj; Zaqout, Sami; Neubert, Gerda; Schindler, Detlev; Ninnemann, Olaf; Gräf, Ralph; Seiler, Andrea EM; Kaindl, Angela M

    2015-01-01

    Biallelic mutations in the gene encoding centrosomal CDK5RAP2 lead to autosomal recessive primary microcephaly (MCPH), a disorder characterized by pronounced reduction in volume of otherwise architectonical normal brains and intellectual deficit. The current model for the microcephaly phenotype in MCPH invokes a premature shift from symmetric to asymmetric neural progenitor-cell divisions with a subsequent depletion of the progenitor pool. The isolated neural phenotype, despite the ubiquitous expression of CDK5RAP2, and reports of progressive microcephaly in individual MCPH cases prompted us to investigate neural and non-neural differentiation of Cdk5rap2-depleted and control murine embryonic stem cells (mESC). We demonstrate an accumulating proliferation defect of neurally differentiating Cdk5rap2-depleted mESC and cell death of proliferative and early postmitotic cells. A similar effect does not occur in non-neural differentiation into beating cardiomyocytes, which is in line with the lack of non-central nervous system features in MCPH patients. Our data suggest that MCPH is not only caused by premature differentiation of progenitors, but also by reduced propagation and survival of neural progenitors. PMID:25942099

  5. Open-Source Wax RepRap 3-D Printer for Rapid Prototyping Paper-Based Microfluidics.

    PubMed

    Pearce, J M; Anzalone, N C; Heldt, C L

    2016-08-01

    The open-source release of self-replicating rapid prototypers (RepRaps) has created a rich opportunity for low-cost distributed digital fabrication of complex 3-D objects such as scientific equipment. For example, 3-D printable reactionware devices offer the opportunity to combine open hardware microfluidic handling with lab-on-a-chip reactionware to radically reduce costs and increase the number and complexity of microfluidic applications. To further drive down the cost while improving the performance of lab-on-a-chip paper-based microfluidic prototyping, this study reports on the development of a RepRap upgrade capable of converting a Prusa Mendel RepRap into a wax 3-D printer for paper-based microfluidic applications. An open-source hardware approach is used to demonstrate a 3-D printable upgrade for the 3-D printer, which combines a heated syringe pump with the RepRap/Arduino 3-D control. The bill of materials, designs, basic assembly, and use instructions are provided, along with a completely free and open-source software tool chain. The open-source hardware device described here accelerates the potential of the nascent field of electrochemical detection combined with paper-based microfluidics by dropping the marginal cost of prototyping to nearly zero while accelerating the turnover between paper-based microfluidic designs.

  6. Role of electrostatics in differential binding of RalGDS to Rap mutations E30D and K31E investigated by vibrational spectroscopy of thiocyanate probes.

    PubMed

    Ragain, Christina M; Newberry, Robert W; Ritchie, Andrew W; Webb, Lauren J

    2012-08-01

    The human protein Rap1A (Rap) is a member of the Ras superfamily of GTPases that binds to the downstream effector Ral guanine nucleotide dissociation stimulator (RalGDS). Although Ras and Rap have nearly identical amino acid sequences and structures along the effector binding surface, the charge reversal mutation Rap K31E has previously been shown to increase the dissociation constant of Rap-RalGDS docking to values similar to that of Ras-RalGDS docking. This indicates that the difference in charge at position 31 could provide a mechanism for Ral to distinguish two structurally similar but functionally distinct GTPases, which would be of vital importance for appropriate biological function. In this report, vibrational Stark effect spectroscopy, dissociation constant measurements, and molecular dynamics simulations were used to investigate the role that electrostatic field differences caused by the charge reversal mutation Rap K31E play in determining the binding specificity of RalGDS to Rap versus Ras. To do this, six variants of RalGDS carrying a thiocyanate electrostatic probe were docked with three Rap mutants, E30D, K31E, and E30D/K31E. The change in absorption energy of the thiocyanate probe caused by RalGDS docking to these Rap variants was then compared to that observed with wild-type Ras. Three trends emerged: the expected reversion behavior, an additive behavior of the two single mutations, and cancelation of the effects of each single mutation in the double mutant. These observations are explained with a physical model of the position of the thiocyanate probe with respect to the mutated residue based on molecular dynamics simulations.

  7. Mutational Analysis Defines a C-Terminal Tail Domain of Rap1 Essential for Telomeric Silencing in Saccharomyces Cerevisiae

    PubMed Central

    Liu, C.; Mao, X.; Lustig, A. J.

    1994-01-01

    Alleles specifically defective in telomeric silencing were generated by in vitro mutagenesis of the yeast RAP1 gene. The most severe phenotypes occur with three mutations in the C-terminal 28 amino acids. Two of the alleles are nonsense mutations resulting in truncated repressor/activator protein 1 (RAP1) species lacking the C-terminal 25-28 amino acids; the third allele is a missense mutation within this region. These alleles define a novel 28-amino acid region, termed the C-terminal tail domain, that is essential for telomeric and HML silencing. Using site-directed mutagenesis, an 8-amino acid region (amino acids 818-825) that is essential for telomeric silencing has been localized within this domain. Further characterization of these alleles has indicated that the C-terminal tail domain also plays a role in telomere size control. The function of the C-terminal tail in telomere maintenance is not mediated through the RAP1 interacting factor RIF1: rap1 alleles defective in both the C-terminal tail and RIF1 interaction domains have additive effects on telomere length. Overproduction of SIR3, a dose-dependent enhancer of telomeric silencing, suppresses the telomeric silencing, but not length, phenotypes of a subset of C-terminal tail alleles. In contrast, an allele that truncates the terminal 28 amino acids of RAP1 is refractory to SIR3 overproduction. These results indicate that the C-terminal tail domain is required for SIR3-dependent enhancement of telomeric silencing. These data also suggest a distinct set of C-terminal requirements for telomere size control and telomeric silencing. PMID:7896088

  8. Typification of virulent and low virulence Babesia bigemina clones by 18S rRNA and rap-1c.

    PubMed

    Thompson, C; Baravalle, M E; Valentini, B; Mangold, A; Torioni de Echaide, S; Ruybal, P; Farber, M; Echaide, I

    2014-06-01

    The population structure of original Babesia bigemina isolates and reference strains with a defined phenotypic profile was assessed using 18S rRNA and rap-1c genes. Two reference strains, BbiS2P-c (virulent) and BbiS1A-c (low virulence), were biologically cloned in vitro. The virulence profile of the strains and clones was assessed in vivo. One fully virulent and one low-virulence clone were mixed in identical proportions to evaluate their growth efficiency in vitro. Each clone was differentiated by two microsatellites and the gene gp45. The 18S rRNA and rap-1c genes sequences from B. bigemina biological clones and their parental strains, multiplied exclusively in vivo or in vitro, were compared with strain JG-29. The virulence of clones derived from the BbiS2P-c strain was variable. Virulent clone Bbi9P1 grew more efficiently in vitro than did the low-virulence clone Bbi2A1. The haplotypes generated by the nucleotide polymorphism, localized in the V4 region of the 18S rRNA, allowed the identification of three genotypes. The rap-1c haplotypes allowed defining four genotypes. Parental and original strains were defined by multiple haplotypes identified in both genes. The rap-1c gene, analyzed by high-resolution melting (HRM), allowed discrimination between two genotypes according to their phenotype, and both were different from JG-29. B. bigemina biological clones made it possible to define the population structure of isolates and strains. The polymorphic regions of the 18S rRNA and rap-1c genes allowed the identification of different subpopulations within original B. bigemina isolates by the definition of several haplotypes and the differentiation of fully virulent from low virulence clones. PMID:24681200

  9. Typification of virulent and low virulence Babesia bigemina clones by 18S rRNA and rap-1c.

    PubMed

    Thompson, C; Baravalle, M E; Valentini, B; Mangold, A; Torioni de Echaide, S; Ruybal, P; Farber, M; Echaide, I

    2014-06-01

    The population structure of original Babesia bigemina isolates and reference strains with a defined phenotypic profile was assessed using 18S rRNA and rap-1c genes. Two reference strains, BbiS2P-c (virulent) and BbiS1A-c (low virulence), were biologically cloned in vitro. The virulence profile of the strains and clones was assessed in vivo. One fully virulent and one low-virulence clone were mixed in identical proportions to evaluate their growth efficiency in vitro. Each clone was differentiated by two microsatellites and the gene gp45. The 18S rRNA and rap-1c genes sequences from B. bigemina biological clones and their parental strains, multiplied exclusively in vivo or in vitro, were compared with strain JG-29. The virulence of clones derived from the BbiS2P-c strain was variable. Virulent clone Bbi9P1 grew more efficiently in vitro than did the low-virulence clone Bbi2A1. The haplotypes generated by the nucleotide polymorphism, localized in the V4 region of the 18S rRNA, allowed the identification of three genotypes. The rap-1c haplotypes allowed defining four genotypes. Parental and original strains were defined by multiple haplotypes identified in both genes. The rap-1c gene, analyzed by high-resolution melting (HRM), allowed discrimination between two genotypes according to their phenotype, and both were different from JG-29. B. bigemina biological clones made it possible to define the population structure of isolates and strains. The polymorphic regions of the 18S rRNA and rap-1c genes allowed the identification of different subpopulations within original B. bigemina isolates by the definition of several haplotypes and the differentiation of fully virulent from low virulence clones.

  10. The de-ubiquitylating enzymes USP26 and USP37 regulate homologous recombination by counteracting RAP80.

    PubMed

    Typas, Dimitris; Luijsterburg, Martijn S; Wiegant, Wouter W; Diakatou, Michaela; Helfricht, Angela; Thijssen, Peter E; van den Broek, Bram; van de Broek, Bram; Mullenders, Leon H; van Attikum, Haico

    2015-08-18

    The faithful repair of DNA double-strand breaks (DSBs) is essential to safeguard genome stability. DSBs elicit a signaling cascade involving the E3 ubiquitin ligases RNF8/RNF168 and the ubiquitin-dependent assembly of the BRCA1-Abraxas-RAP80-MERIT40 complex. The association of BRCA1 with ubiquitin conjugates through RAP80 is known to be inhibitory to DSB repair by homologous recombination (HR). However, the precise regulation of this mechanism remains poorly understood. Through genetic screens we identified USP26 and USP37 as key de-ubiquitylating enzymes (DUBs) that limit the repressive impact of RNF8/RNF168 on HR. Both DUBs are recruited to DSBs where they actively remove RNF168-induced ubiquitin conjugates. Depletion of USP26 or USP37 disrupts the execution of HR and this effect is alleviated by the simultaneous depletion of RAP80. We demonstrate that USP26 and USP37 prevent excessive spreading of RAP80-BRCA1 from DSBs. On the other hand, we also found that USP26 and USP37 promote the efficient association of BRCA1 with PALB2. This suggests that these DUBs limit the ubiquitin-dependent sequestration of BRCA1 via the BRCA1-Abraxas-RAP80-MERIT40 complex, while promoting complex formation and cooperation of BRCA1 with PALB2-BRCA2-RAD51 during HR. These findings reveal a novel ubiquitin-dependent mechanism that regulates distinct BRCA1-containing complexes for efficient repair of DSBs by HR. PMID:26101254

  11. Neural Correlates of Lyrical Improvisation: An fMRI Study of Freestyle Rap

    PubMed Central

    Liu, Siyuan; Chow, Ho Ming; Xu, Yisheng; Erkkinen, Michael G.; Swett, Katherine E.; Eagle, Michael W.; Rizik-Baer, Daniel A.; Braun, Allen R.

    2012-01-01

    The neural correlates of creativity are poorly understood. Freestyle rap provides a unique opportunity to study spontaneous lyrical improvisation, a multidimensional form of creativity at the interface of music and language. Here we use functional magnetic resonance imaging to characterize this process. Task contrast analyses indicate that improvised performance is characterized by dissociated activity in medial and dorsolateral prefrontal cortices, providing a context in which stimulus-independent behaviors may unfold in the absence of conscious monitoring and volitional control. Connectivity analyses reveal widespread improvisation-related correlations between medial prefrontal, cingulate motor, perisylvian cortices and amygdala, suggesting the emergence of a network linking motivation, language, affect and movement. Lyrical improvisation appears to be characterized by altered relationships between regions coupling intention and action, in which conventional executive control may be bypassed and motor control directed by cingulate motor mechanisms. These functional reorganizations may facilitate the initial improvisatory phase of creative behavior. PMID:23155479

  12. Data-Rich Astronomy: Mining Sky Surveys with PhotoRApToR

    NASA Astrophysics Data System (ADS)

    Cavuoti, Stefano; Brescia, Massimo; Longo, Giuseppe

    2014-05-01

    In the last decade a new generation of telescopes and sensors has allowed the production of a very large amount of data and astronomy has become a data-rich science. New automatic methods largely based on machine learning are needed to cope with such data tsunami. We present some results in the fields of photometric redshifts and galaxy classification, obtained using the MLPQNA algorithm available in the DAMEWARE (Data Mining and Web Application Resource) for the SDSS galaxies (DR9 and DR10). We present PhotoRApToR (Photometric Research Application To Redshift): a Java based desktop application capable to solve regression and classification problems and specialized for photo-z estimation.

  13. Immunohistochemical analysis of cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA) in murine, canine, bovine and equine cerebrospinal tissues.

    PubMed

    Tokunaga, Satoshi; Fujiki, Makoto; Yabuki, Akira; Misumi, Kazuhiro

    2012-04-01

    Cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA), which appears abundantly in hypertrophic cartilage at the stage of endochondral ossification, is also detected in cerebrospinal fluid (CSF) following spinal cord injury. In this study, the localization of the CD-RAP/MIA molecule in normal tissues of the spine and brain obtained from mice, rats, dogs, cattle and horses was examined using immunohistochemistry with a specific antibody. The positive signals of CD-RAP/MIA were found at nerve cells in the spinal cords of all species and were especially strong at cerebellar Purkinje cells. The results suggested that CD-RAP/MIA included in normal cerebrospinal tissues could be a biomarker associated with tissue injuries, as the molecules might flow into the CSF.

  14. Detection of Differentially Expressed Genes in an Isogenic Breast Metastasis Model using RNA Arbitrarily Primed-Polymerase Chain Reaction Coupled with Array Hybridization (RAP-Array)

    PubMed Central

    Sloan, Derek D.; Nicholson, Ben; Urquidi, Virginia; Goodison, Steve

    2004-01-01

    To facilitate the study of the mechanisms of breast cancer metastasis we have previously characterized a pair of breast tumor cell lines that originate from the same breast tumor cell line MDA-MB-435, but which have diametrically opposite metastatic capabilities. These cell lines constitute a stable and accessible experimental system for the identification of metastasis-related genes and for the study of their role in the process of metastasis. In this study, we used a combination of RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) fingerprinting and cDNA arrays (here termed “RAP-array”) to identify genes differentially expressed with respect to metastatic phenotype. RAP-PCR was used to generate radioactive probes of reduced complexity for hybridization to nylon membranes containing 588 cDNAs of known identity. Single RAP-PCR fingerprint probes hybridized from 61 (10.4%) to 116 (19.7%) of the filter array targets, with a signal detection overlap of ∼21%. A total of 344 (57%) of the 588 target genes were detected by five single RAP-PCR fingerprints. The advantage of using reduced complexity probes was highlighted by the fact that the combination of RAP probes before hybridization compromised the overall detection rate by up to 40%. Sequential application of RAP-PCR probes allowed the screening of a greater, and an alternative fraction of the transcript population than was achieved with a radiolabeled total cDNA probe. Verification by quantitative reverse transcriptase-PCR confirmed significantly increased expression of keratin 9 (>100-fold) in nonmetastatic breast tumor cells and of CD70 (fivefold) in metastatic cells. The differential expression of keratin 9 and CD70 was maintained between cells grown as primary xenografts in athymic mice. The RAP-array method enabled the detection of genes not revealed using other screening methods and that are candidates for further investigation in the context of metastatic phenotype. PMID:14695344

  15. Rap1 induces cytokine production in pro-inflammatory macrophages through NFκB signaling and is highly expressed in human atherosclerotic lesions

    PubMed Central

    Cai, Yin; Sukhova, Galina K; Wong, Hoi Kin; Xu, Aimin; Tergaonkar, Vinay; Vanhoutte, Paul M; Tang, Eva Hoi Ching

    2015-01-01

    Repressor activator protein 1 (Rap1) is essential for maintaining telomere length and structural integrity, but it also exerts other non-telomeric functions. The present study tested the hypothesis that Rap1 is released into the cytoplasm and induces production of pro-inflammatory cytokines via nuclear factor kappa B (NFκB) signaling in macrophages, a cell type involved in the development and progression of atherosclerotic lesions. Western blotting analysis confirmed that Rap1 was present in the cytoplasm of differentiated human monocytic leukemia cells (THP-1, a macrophage-like cell line). Co-immunoprecipitation assay revealed a direct interaction between Rap1 and I kappa B kinase (IKK). Knockdown of Rap1 suppressed lipopolysaccharide-mediated activation of NFκB, and phosphorylation of inhibitor of kappa B α (IκBα) and p65 in THP-1 macrophages. The reduction of NFκB activity was paralleled by a decreased production of NFκB-dependent pro-inflammatory cytokines and an increased expression of IκBα (native NFκB inhibitor) in various macrophage models with pro-inflammatory phenotype, including THP-1, mouse peritoneal macrophages and bone marrow-derived M1 macrophages. These changes were observed selectively in pro-inflammatory macrophages but not in bone marrow-derived M2 macrophages (with an anti-inflammatory phenotype), mouse lung endothelial cells, human umbilical vein endothelial cells or human aortic smooth muscle cells. Immunostaining revealed that Rap1 was localized mainly in macrophage-rich areas in human atherosclerotic plaques and that the presence of Rap1 was positively correlated with the advancement of the disease process. In pro-inflammatory macrophages, Rap1 promotes cytokine production via NFκB activation favoring a pro-inflammatory environment which may contribute to the development and progression of atherosclerosis. PMID:26505215

  16. Effects of prostaglandin E{sub 2} on the subcellular localization of Epac-1 and Rap1 proteins during Fc{gamma}-receptor-mediated phagocytosis in alveolar macrophages

    SciTech Connect

    Brock, Thomas G.; Serezani, Carlos H.; Carstens, Jennifer K.; Peters-Golden, Marc; Aronoff, David M.

    2008-01-15

    Recent studies have demonstrated a central role for the exchange protein activated by cAMP (Epac) in the inhibition of Fc{gamma}-receptor-mediated phagocytosis and bacterial killing by prostaglandin E{sub 2} (PGE{sub 2}) in macrophages. However, the subcellular localization of Epac, and its primary target Rap1, has yet to be determined in primary macrophages. Therefore, we used immunofluorescent techniques and phagosome isolation to localize Epac-1 and Rap1 in alveolar macrophages. Epac-1 was predominantly expressed on punctate and tubular membranes throughout the cell body; on the plasma membrane; and co-localized with microtubule organizing centers (MTOCs). Rap1 was abundant on punctate membranes, less abundant on plasma membrane, and also found on MTOCs. Following PGE{sub 2} treatment, Epac-1, but not Rap1, accumulated on the nuclear envelope and disappeared from MTOCs. By immunofluorescent microscopy, both Epac-1 and Rap1 were seen to associate with phagosomes containing IgG-opsonized beads, but this association appeared weak, as we failed to observe such interactions in phagosomes isolated from cells at various time points after bead ingestion. Strikingly, however, Epac-1, but not Rap1, appeared to accumulate on maturing phagosomes, but only after PGE{sub 2} treatment (or treatment with a selective Epac-1 agonist). This association was confirmed in isolated phagosome preparations. The changes in Epac-1 localization were too slow to account for the inhibitory effects of PGE{sub 2} on phagocytosis. However, the appearance of Epac-1 on late phagosomes following PGE{sub 2} treatment might be important for suppressing H{sub 2}O{sub 2} production and inhibiting the killing of intraphagosomal pathogens. The absence of Rap1 on late phagosomes suggests that the effect of Epac-1 might not require Rap1.

  17. NMR structure of a complex containing the TFIIF subunit RAP74 and the RNA polymerase II carboxyl-terminal domain phosphatase FCP1

    PubMed Central

    Nguyen, Bao D.; Abbott, Karen L.; Potempa, Krzysztof; Kobor, Michael S.; Archambault, Jacques; Greenblatt, Jack; Legault, Pascale; Omichinski, James G.

    2003-01-01

    FCP1 [transcription factor IIF (TFIIF)-associated carboxyl-terminal domain (CTD) phosphatase] is the only identified phosphatase specific for the phosphorylated CTD of RNA polymerase II (RNAP II). The phosphatase activity of FCP1 is enhanced in the presence of the large subunit of TFIIF (RAP74 in humans). It has been demonstrated that the CTD of RAP74 (cterRAP74; residues 436–517) directly interacts with the highly acidic CTD of FCP1 (cterFCP; residues 879–961 in human). In this manuscript, we have determined a high-resolution solution structure of a cterRAP74/cterFCP complex by NMR spectroscopy. Interestingly, the cterFCP protein is completely disordered in the unbound state, but forms an α-helix (H1′; E945–M961) in the complex. The cterRAP74/cterFCP binding interface relies extensively on van der Waals contacts between hydrophobic residues from the H2 and H3 helices of cterRAP74 and hydrophobic residues from the H1′ helix of cterFCP. The binding interface also contains two critical electrostatic interactions involving aspartic acid residues from H1′ of cterFCP and lysine residues from both H2 and H3 of cterRAP74. There are also three additional polar interactions involving highly conserved acidic residues from the H1′ helix. The cterRAP74/cterFCP complex is the first high-resolution structure between an acidic residue-rich domain from a holoenzyme-associated regulatory protein and a general transcription factor. The structure defines a clear role for both hydrophobic and acidic residues in protein/protein complexes involving acidic residue-rich domains in transcription regulatory proteins. PMID:12732728

  18. NMR structure of a complex containing the TFIIF subunit RAP74 and the RNA polymerase II carboxyl-terminal domain phosphatase FCP1.

    PubMed

    Nguyen, Bao D; Abbott, Karen L; Potempa, Krzysztof; Kobor, Michael S; Archambault, Jacques; Greenblatt, Jack; Legault, Pascale; Omichinski, James G

    2003-05-13

    FCP1 [transcription factor IIF (TFIIF)-associated carboxyl-terminal domain (CTD) phosphatase] is the only identified phosphatase specific for the phosphorylated CTD of RNA polymerase II (RNAP II). The phosphatase activity of FCP1 is enhanced in the presence of the large subunit of TFIIF (RAP74 in humans). It has been demonstrated that the CTD of RAP74 (cterRAP74; residues 436-517) directly interacts with the highly acidic CTD of FCP1 (cterFCP; residues 879-961 in human). In this manuscript, we have determined a high-resolution solution structure of a cterRAP74cterFCP complex by NMR spectroscopy. Interestingly, the cterFCP protein is completely disordered in the unbound state, but forms an alpha-helix (H1'; E945-M961) in the complex. The cterRAP74cterFCP binding interface relies extensively on van der Waals contacts between hydrophobic residues from the H2 and H3 helices of cterRAP74 and hydrophobic residues from the H1' helix of cterFCP. The binding interface also contains two critical electrostatic interactions involving aspartic acid residues from H1' of cterFCP and lysine residues from both H2 and H3 of cterRAP74. There are also three additional polar interactions involving highly conserved acidic residues from the H1' helix. The cterRAP74cterFCP complex is the first high-resolution structure between an acidic residue-rich domain from a holoenzyme-associated regulatory protein and a general transcription factor. The structure defines a clear role for both hydrophobic and acidic residues in proteinprotein complexes involving acidic residue-rich domains in transcription regulatory proteins. PMID:12732728

  19. Mutation of Dcdc2 in mice leads to impairments in auditory processing and memory ability.

    PubMed

    Truong, D T; Che, A; Rendall, A R; Szalkowski, C E; LoTurco, J J; Galaburda, A M; Holly Fitch, R

    2014-11-01

    Dyslexia is a complex neurodevelopmental disorder characterized by impaired reading ability despite normal intellect, and is associated with specific difficulties in phonological and rapid auditory processing (RAP), visual attention and working memory. Genetic variants in Doublecortin domain-containing protein 2 (DCDC2) have been associated with dyslexia, impairments in phonological processing and in short-term/working memory. The purpose of this study was to determine whether sensory and behavioral impairments can result directly from mutation of the Dcdc2 gene in mice. Several behavioral tasks, including a modified pre-pulse inhibition paradigm (to examine auditory processing), a 4/8 radial arm maze (to assess/dissociate working vs. reference memory) and rotarod (to examine sensorimotor ability and motor learning), were used to assess the effects of Dcdc2 mutation. Behavioral results revealed deficits in RAP, working memory and reference memory in Dcdc2(del2/del2) mice when compared with matched wild types. Current findings parallel clinical research linking genetic variants of DCDC2 with specific impairments of phonological processing and memory ability.

  20. Attenuation of rodent neuropathic pain by an orally active peptide, RAP-103, which potently blocks CCR2- and CCR5-mediated monocyte chemotaxis and inflammation.

    PubMed

    Padi, Satyanarayana S V; Shi, Xiang Q; Zhao, Yuan Q; Ruff, Michael R; Baichoo, Noel; Pert, Candace B; Zhang, Ji

    2012-01-01

    Chemokine signaling is important in neuropathic pain, with microglial cells expressing CCR2 playing a well-established key role. DAPTA, a HIV gp120-derived CCR5 entry inhibitor, has been shown to inhibit CCR5-mediated monocyte migration and to attenuate neuroinflammation. We report here that as a stabilized analog of DAPTA, the short peptide RAP-103 exhibits potent antagonism for both CCR2 (half maximal inhibitory concentration [IC50] 4.2 pM) and CCR5 (IC50 0.18 pM) in monocyte chemotaxis. Oral administration of RAP-103 (0.05-1 mg/kg) for 7 days fully prevents mechanical allodynia and inhibits the development of thermal hyperalgesia after partial ligation of the sciatic nerve in rats. Administered from days 8 to 12, RAP-103 (0.2-1 mg/kg) reverses already established hypersensitivity. RAP-103 relieves behavioral hypersensitivity, probably through either or both CCR2 and CCR5 blockade, because by using genetically deficient animals, we demonstrated that in addition to CCR2, CCR5 is also required for the development of neuropathic pain. Moreover, RAP-103 is able to reduce spinal microglial activation and monocyte infiltration, and to inhibit inflammatory responses evoked by peripheral nerve injury that cause chronic pain. Our findings suggest that targeting CCR2/CCR5 should provide greater efficacy than targeting CCR2 or CCR5 alone, and that dual CCR2/CCR5 antagonist RAP-103 has the potential for broad clinical use in neuropathic pain treatment.

  1. The first case of CDK5RAP2-related primary microcephaly in a non-consanguineous patient identified by next generation sequencing.

    PubMed

    Tan, Christopher A; Topper, Scott; Ward Melver, Catherine; Stein, Jennifer; Reeder, Amanda; Arndt, Kelly; Das, Soma

    2014-04-01

    Primary autosomal recessive microcephaly (MCPH) is a genetically heterogeneous condition characterized by congenital microcephaly and intellectual disability. To date, 10 MCPH loci have been identified and due to the genetic heterogeneity of this condition, molecular testing for MCPH can be complicated. Our methods involved employing a next generation sequencing panel of MCPH-related genes allowing for the evaluation of multiple disease loci simultaneously. Next generation sequencing analysis of a 6 year old female with primary microcephaly identified novel compound heterozygous mutations (c.524_528del and c.4005-1G>A) in the CDK5RAP2 gene. A review of the published literature to date reveals that only three mutations have been previously reported in the CDK5RAP2 gene in the homozygous state in three Northern Pakistani and one Somali consanguineous MCPH families. Our patient represents the first non-consanguineous Caucasian individual to have been identified with CDK5RAP2-related MCPH. As only a handful of patients have been reported in the literature with CDK5RAP2-related MCPH, we anticipate the identification of individuals with CDK5RAP2 mutations from all ethnic backgrounds will continue. Our patient contributes to the ethnic and genotypic spectrum of CDK5RAP2-related MCPH and supports the occurrence of this genetic condition beyond that of consanguineous families of certain ethnic populations. Our results also highlight the utility of multi-gene sequencing panels to elucidate the etiology of genetically heterogeneous conditions. PMID:23726037

  2. Structural Basis for Small G Protein Effector Interaction of Ras-related Protein 1 (Rap1) and Adaptor Protein Krev Interaction Trapped 1 (KRIT1)

    SciTech Connect

    Li, Xiaofeng; Zhang, Rong; Draheim, Kyle M.; Liu, Weizhi; Calderwood, David A.; Boggon, Titus J.

    2012-09-17

    Cerebral cavernous malformations (CCMs) affect 0.1-0.5% of the population resulting in leaky vasculature and severe neurological defects. KRIT1 (Krev interaction trapped-1) mutations associate with {approx}40% of familial CCMs. KRIT1 is an effector of Ras-related protein 1 (Rap1) GTPase. Rap1 relocalizes KRIT1 from microtubules to cell membranes to impact integrin activation, potentially important for CCM pathology. We report the 1.95 {angstrom} co-crystal structure of KRIT1 FERM domain in complex with Rap1. Rap1-KRIT1 interaction encompasses an extended surface, including Rap1 Switch I and II and KRIT1 FERM F1 and F2 lobes. Rap1 binds KRIT1-F1 lobe using a GTPase-ubiquitin-like fold interaction but binds KRIT1-F2 lobe by a novel interaction. Point mutagenesis confirms the interaction. High similarity between KRIT1-F2/F3 and talin is revealed. Additionally, the mechanism for FERM domains acting as GTPase effectors is suggested. Finally, structure-based alignment of each lobe suggests classification of FERM domains as ERM-like and TMFK-like (talin-myosin-FAK-KRIT-like) and that FERM lobes resemble domain 'modules.'

  3. A demonstration of the applicability of implementing the enhanced Remedial Action Priority System (RAPS) for environmental releases

    SciTech Connect

    Whelan, G.; Droppo, J.G. Jr.; Strenge, D.L.; Walter, M.B.; Buck, J.W.

    1989-12-01

    The Remedial Action Priority System (RAPS) and the Multimedia Environmental Pollutant Assessment System (MEPAS) were developed to prioritize problems associated with potential releases of hazardous chemical and radioactive materials in a scientific and objective manner based on limited site information. This report documents the model testing efforts of the RAPS/MEPAS methodology for the atmospheric, surface water, groundwater, and exposure components. Comparisons are given of model outputs with measured data at three sites: the US Department of Energy's Mound facility in Ohio and Hanford facility in Washington, and a chromium-cadmium plating site in New York. The results show that the simulated magnitudes, spacial and temporal trends, and distributions of contaminants corresponded well with the measured data. 25 refs., 86 figs., 26 tabs.

  4. Binding site structure of one LRP-RAP complex: implications for a common ligand-receptor binding motif.

    PubMed

    Jensen, Gitte A; Andersen, Olav M; Bonvin, Alexandre M J J; Bjerrum-Bohr, Ida; Etzerodt, Michael; Thøgersen, Hans C; O'Shea, Charlotte; Poulsen, Flemming M; Kragelund, Birthe B

    2006-09-29

    The low-density lipoprotein receptor-related protein (LRP) interacts with more than 30 ligands of different sizes and structures that can all be replaced by the receptor-associated protein (RAP). The double module of complement type repeats, CR56, of LRP binds many ligands including all three domains of RAP and alpha2-macroglobulin, which promotes the catabolism of the Abeta-peptide implicated in Alzheimer's disease. To understand the receptor-ligand cross-talk, the NMR structure of CR56 has been solved and ligand binding experiments with RAP domain 1 (RAPd1) have been performed. From chemical shift perturbations of both binding partners upon complex formation, a HADDOCK model of the complex between CR56 and RAPd1 has been obtained. The binding residues are similar to a common binding motif suggested from alpha2-macroglobulin binding studies and provide evidence for an understanding of their mutual cross-competition pattern. The present structural results convey a simultaneous description of both binding partners of an LRP-ligand complex and open a route to a broader understanding of the binding specificity of the LRP receptor, which may involve a general four-residue receptor-ligand recognition motif common to all LRP ligands. The present result may be beneficial in the design of antagonists of ligand binding to the LDL receptor family, and especially of drugs for treatment of Alzheimer's disease.

  5. Streptococcus pneumoniae ClpL Modulates Adherence to A549 Human Lung Cells through Rap1/Rac1 Activation

    PubMed Central

    Nguyen, Cuong Thach; Le, Nhat-Tu; Tran, Thao Dang-Hien; Kim, Eun-Hye; Park, Sang-Sang; Luong, Truc Thanh; Chung, Kyung-Tae; Pyo, Suhkneung

    2014-01-01

    Caseinolytic protease L (ClpL) is a member of the HSP100/Clp chaperone family, which is found mainly in Gram-positive bacteria. ClpL is highly expressed during infection for refolding of stress-induced denatured proteins, some of which are important for adherence. However, the role of ClpL in modulating pneumococcal virulence is poorly understood. Here, we show that ClpL impairs pneumococcal adherence to A549 lung cells by inducing and activating Rap1 and Rac1, thus increasing phosphorylation of cofilin (inactive form). Moreover, infection with a clpL mutant (ΔclpL) causes a greater degree of filopodium formation than D39 wild-type (WT) infection. Inhibition of Rap1 and Rac1 impairs filopodium formation and pneumococcal adherence. Therefore, ClpL can reduce pneumococcal adherence to A549 cells, likely via modulation of Rap1- and Rac1-mediated filopodium formation. These results demonstrate a potential role for ClpL in pneumococcal resistance to host cell adherence during infection. This study provides insight into further understanding the interactions between hosts and pathogens. PMID:24980975

  6. GCR1, a transcriptional activator in Saccharomyces cerevisiae, complexes with RAP1 and can function without its DNA binding domain.

    PubMed Central

    Tornow, J; Zeng, X; Gao, W; Santangelo, G M

    1993-01-01

    In Saccharomyces cerevisiae, efficient expression of glycolytic and translational component genes requires two DNA binding proteins, RAP1 (which binds to UASRPG) and GCR1 (which binds to the CT box). We generated deletions in GCR1 to test the validity of several different models for GCR1 function. We report here that the C-terminal half of GCR1, which includes the domain required for DNA binding to the CT box in vitro, can be removed without affecting GCR1-dependent transcription of either the glycolytic gene ADH1 or the translational component genes TEF1 and TEF2. We have also identified an activation domain within a segment of the GCR1 protein (the N-terminal third) that is essential for in vivo function. RAP1 and GCR1 can be co-immunoprecipitated from whole cell extracts, suggesting that they form a complex in vivo. The data are most consistent with a model in which GCR1 is attracted to DNA through contact with RAP1. Images PMID:8508768

  7. Phloretin exhibits an anticancer effect and enhances the anticancer ability of cisplatin on non-small cell lung cancer cell lines by regulating expression of apoptotic pathways and matrix metalloproteinases.

    PubMed

    Ma, Lijie; Wang, Ruixuan; Nan, Yandong; Li, Wangping; Wang, Qingwei; Jin, Faguang

    2016-02-01

    Non-small cell lung cancer (NSCLC) accounts for 80-85% of all lung cancer cases and the prognosis of NSCLC patients is unsatisfactory since 5-year survival rate of NSCLC is still as low as 11%. Natural compounds derived from plants with few or no side effects have been recognized as alternative or auxiliary cure for cancer patients. Phloretin is such an agent possessing various pharmacological activities; however, there is scarce information on its anticancer effects on NSCLC. It was evaluated and confirmed, in the present study, that phloretin inhibited proliferation and induced apoptosis in A549, Calu-1, H838 and H520 cells in a dose-dependent manner, phloretin also suppressed the invasion and migration of NSCLC cells. We further confirmed that phloretin dose-dependently suppressed the expression of Bcl-2, increased the protein expression of cleaved-caspase-3 and -9, and deregulated the expression of matrix metalloproteinases (MMP)-2 and -9 on gene and protein levels. Besides, evaluations revealed that phloretin enhanced the anticancer effects of cisplatin on inhibition of proliferation and induction of apoptosis in NSCLC cells. Moreover, phloretin facilitated the effects of cisplatin on deregulation of Bcl-2, MMP-2 and -9, and upregulation of cleaved-caspase-3 and -9. In conclusion, the present study demonstrated that phloretin possessed anticancer effects and enhanced the anticancer effects of cisplatin on NSCLC cell lines by suppressing proliferation, inducing apoptosis and inhibiting invasion and migration of the cells through regulating apoptotic pathways and MMPs. PMID:26692364

  8. Phloretin exhibits an anticancer effect and enhances the anticancer ability of cisplatin on non-small cell lung cancer cell lines by regulating expression of apoptotic pathways and matrix metalloproteinases.

    PubMed

    Ma, Lijie; Wang, Ruixuan; Nan, Yandong; Li, Wangping; Wang, Qingwei; Jin, Faguang

    2016-02-01

    Non-small cell lung cancer (NSCLC) accounts for 80-85% of all lung cancer cases and the prognosis of NSCLC patients is unsatisfactory since 5-year survival rate of NSCLC is still as low as 11%. Natural compounds derived from plants with few or no side effects have been recognized as alternative or auxiliary cure for cancer patients. Phloretin is such an agent possessing various pharmacological activities; however, there is scarce information on its anticancer effects on NSCLC. It was evaluated and confirmed, in the present study, that phloretin inhibited proliferation and induced apoptosis in A549, Calu-1, H838 and H520 cells in a dose-dependent manner, phloretin also suppressed the invasion and migration of NSCLC cells. We further confirmed that phloretin dose-dependently suppressed the expression of Bcl-2, increased the protein expression of cleaved-caspase-3 and -9, and deregulated the expression of matrix metalloproteinases (MMP)-2 and -9 on gene and protein levels. Besides, evaluations revealed that phloretin enhanced the anticancer effects of cisplatin on inhibition of proliferation and induction of apoptosis in NSCLC cells. Moreover, phloretin facilitated the effects of cisplatin on deregulation of Bcl-2, MMP-2 and -9, and upregulation of cleaved-caspase-3 and -9. In conclusion, the present study demonstrated that phloretin possessed anticancer effects and enhanced the anticancer effects of cisplatin on NSCLC cell lines by suppressing proliferation, inducing apoptosis and inhibiting invasion and migration of the cells through regulating apoptotic pathways and MMPs.

  9. Interleukin 18 augments growth ability via NF-κB and p38/ATF2 pathways by targeting cyclin B1, cyclin B2, cyclin A2, and Bcl-2 in BRL-3A rat liver cells.

    PubMed

    Zhang, Jihong; Pan, Cuiyun; Xu, Tiantian; Niu, Zhipeng; Ma, Chengkai; Xu, Cunshuan

    2015-05-25

    Interleukin 18 (IL-18) is a pleiotropic cytokine and capable of stimulating proliferation of certain cell types. Nonetheless, its effect on normal liver cells cultured remains unclear. In the present study, we discovered that IL-18 expression level was remarkably elevated at 3.3 and 8.6h after synchronized BRL-3A rat liver cells (G0 phase) re-entering the cell cycle. In addition, recombinant rat IL-18 (rrIL-18) at dosages 5-10 ng/ml increased the cell viability compared to untreated cells (with medium only) at 24 and 48 h (P<0.05). At the same time, the percentage of BrdU-labeling cells was also significantly increased (P<0.01). On the other hand, knockdown of IL-18 expression with short interference RNA (siRNA), the cell viability began to decline at 24h and significantly decreased compared to negative control (NC) at 48 and 72 h after transfection (P<0.05). Meanwhile, the number of cells in division phase (G2/M) was reduced in parallel. Further, after treatment with rrIL-18 (5 ng/ml), IL-18 and its receptor subunit IL-18Rα increased both at mRNA and protein levels. Moreover, the expression levels of adaptor molecule MyD88, transcription factor NF-κB and its downstream targets cyclin B1 and cyclin B2 were remarkably enhanced in BRL-3A cells stimulated by rrIL-18. Furthermore, transcription factor ATF2 and its targeted genes cyclin A2, Bcl-2 were also markedly increased after treatment with rrIL-18. These results demonstrated that IL-18 can augment cell proliferation via NF-κB and p38/ATF2 pathway by targeting cyclin B1, cyclin B2, cyclin A2 and Bcl-2 in BRL-3A rat liver cells.

  10. Comprehensive chemical characterization of Rapé tobacco products: Nicotine, un-ionized nicotine, tobacco-specific N'-nitrosamines, polycyclic aromatic hydrocarbons, and flavor constituents.

    PubMed

    Stanfill, Stephen B; Oliveira da Silva, André Luiz; Lisko, Joseph G; Lawler, Tameka S; Kuklenyik, Peter; Tyx, Robert E; Peuchen, Elizabeth H; Richter, Patricia; Watson, Clifford H

    2015-08-01

    Rapé, a diverse group of smokeless tobacco products indigenous to South America, is generally used as a nasal snuff and contains substantial amount of plant material with or without tobacco. Previously uncharacterized, rapé contains addictive and harmful chemicals that may have public health implications for users. Here we report % moisture, pH, and the levels of total nicotine, un-ionized nicotine, flavor-related compounds, tobacco-specific N-nitrosamines (TSNAs) and polycyclic aromatic hydrocarbons (PAHs) for manufactured and hand-made rapé. Most rapé products were mildly acidic (pH 5.17-6.23) with total nicotine ranging from 6.32 to 47.6 milligram per gram of sample (mg/g). Calculated un-ionized nicotine ranged from 0.03 to 18.5 mg/g with the highest values associated with hand-made rapés (pH 9.75-10.2), which contain alkaline ashes. In tobacco-containing rapés, minor alkaloid levels and Fourier transform infrared spectra were used to confirm the presence of Nicotiana rustica, a high nicotine tobacco species. There was a wide concentration range of TSNAs and PAHs among the rapés analyzed. Several TSNAs and PAHs identified in the products are known or probable carcinogens according to the International Agency for Research on Cancer. Milligram quantities of some non-tobacco constituents, such as camphor, coumarin, and eugenol, warrant additional evaluation.

  11. RAP: Accurate and Fast Motif Finding Based on Protein-Binding Microarray Data

    PubMed Central

    Orenstein, Yaron; Mick, Eran

    2013-01-01

    Abstract The novel high-throughput technology of protein-binding microarrays (PBMs) measures binding intensity of a transcription factor to thousands of DNA probe sequences. Several algorithms have been developed to extract binding-site motifs from these data. Such motifs are commonly represented by positional weight matrices. Previous studies have shown that the motifs produced by these algorithms are either accurate in predicting in vitro binding or similar to previously published motifs, but not both. In this work, we present a new simple algorithm to infer binding-site motifs from PBM data. It outperforms prior art both in predicting in vitro binding and in producing motifs similar to literature motifs. Our results challenge previous claims that motifs with lower information content are better models for transcription-factor binding specificity. Moreover, we tested the effect of motif length and side positions flanking the “core” motif in the binding site. We show that side positions have a significant effect and should not be removed, as commonly done. A large drop in the results quality of all methods is observed between in vitro and in vivo binding prediction. The software is available on acgt.cs.tau.ac.il/rap. PMID:23464877

  12. NMR structure of a complex formed by the carboxyl-terminal domain of human RAP74 and a phosphorylated peptide from the central domain of the FCP1 phosphatase.

    PubMed

    Yang, Ao; Abbott, Karen L; Desjardins, Alexandre; Di Lello, Paola; Omichinski, James G; Legault, Pascale

    2009-03-10

    Recycling of RNA polymerase II (RNAPII) requires dephosphorylation of the C-terminal domain (CTD) of the largest subunit of the polymerase. FCP1 enables the recycling of RNAPII via its CTD-specific phosphatase activity, which is stimulated by the RAP74 subunit of the general transcription factor TFIIF. Both the central (centFCP1) and C-terminal (cterFCP1) domains of FCP1 interact independently and specifically with the C-terminal domain of RAP74 (cterRAP74), suggesting that these interactions mediate the stimulatory effect of TFIIF on the CTD phosphatase activity of FCP1. Phosphorylation of FCP1 by casein kinase 2 on residues in its central (T584) and C-terminal (S942 and S944) domains stimulates its binding to RAP74 and its CTD phosphatase activity. To improve our understanding of the FCP1-RAP74 interactions, we previously determined the NMR structure of a complex formed by human cterRAP74 and cterFCP1. We now present the high-resolution NMR structure and thermodynamic characterization by isothermal titration calorimetry of a complex formed by the same cterRAP74 domain and a phosphorylated peptide from the central domain of human FCP1 (centFCP1-PO(4)). Comparison of the cterFCP1-cterRAP74 and centFCP1-PO(4)-cterRAP74 complexes indicates that centFCP1 and cterFCP1 both utilize hydrophobic and acidic residues to recognize the same groove of RAP74, but there are significant differences in the details of their interactions. These differences point to the adaptability of RAP74 to recognize the two regions of FCP1. Our NMR and thermodynamic studies further elucidate the complex molecular mechanism by which TFIIF and FCP1 cooperate for RNAPII recycling. PMID:19215094

  13. RasGRP3 limits Toll-like receptor-triggered inflammatory response in macrophages by activating Rap1 small GTPase

    PubMed Central

    Tang, Songqing; Chen, Taoyong; Yu, Zhou; Zhu, Xuhui; Yang, Mingjin; Xie, Bin; Li, Nan; Cao, Xuetao; Wang, Jianli

    2014-01-01

    Host immune cells can detect and destruct invading pathogens via pattern-recognition receptors. Small Rap GTPases act as conserved molecular switches coupling extracellular signals to various cellular responses, but their roles as regulators in Toll-like receptor (TLR) signalling have not been fully elucidated. Here we report that Ras guanine nucleotide-releasing protein 3 (RasGRP3), a guanine nucleotide-exchange factor activating Ras and Rap1, limits production of proinflammatory cytokines (especially IL-6) in macrophages by activating Rap1 on activation by low levels of TLR agonists. We demonstrate that RasGRP3, a dominant member of RasGRPs in macrophages, impairs TLR3/4/9-induced IL-6 production and relieves dextrane sulphate sodium-induced colitis and collagen-induced arthritis. In RasGRP3-deficient RAW264.7 cells obtained by CRISPR-Cas9 genome editing, TLR3/4/9-induced activation of Rap1 was inhibited while ERK1/2 activation was enhanced. Our study suggests that RasGRP3 limits inflammatory response by activating Rap1 on low-intensity pathogen infection, setting a threshold for preventing excessive inflammatory response. PMID:25118589

  14. Rhoptry-associated protein (rap-1) genes in the sheep pathogen Babesia sp. Xinjiang: Multiple transcribed copies differing by 3' end repeated sequences.

    PubMed

    Niu, Qingli; Marchand, Jordan; Yang, Congshan; Bonsergent, Claire; Guan, Guiquan; Yin, Hong; Malandrin, Laurence

    2015-07-30

    Sheep babesiosis occurs mainly in tropical and subtropical areas. The sheep parasite Babesia sp. Xinjiang is widespread in China, and our goal is to characterize rap-1 (rhoptry-associated protein 1) gene diversity and expression as a first step of a long term goal aiming at developing a recombinant subunit vaccine. Seven different rap-1a genes were amplified in Babesia sp. Xinjiang, using degenerate primers designed from conserved motifs. Rap-1b and rap-1c gene types could not be identified. In all seven rap-1a genes, the 5' regions exhibited identical sequences over 936 nt, and the 3' regions differed at 28 positions over 147 nt, defining two types of genes designated α and β. The remaining 3' part varied from 72 to 360 nt in length, depending on the gene. This region consists of a succession of two to ten 36 nt repeats, which explains the size differences. Even if the nucleotide sequences varied, 6 repeats encoded the same stretch of amino acids. Transcription of at least four α and two β genes was demonstrated by standard RT-PCR.

  15. The rap and hor proteins of Erwinia, Serratia and Yersinia: a novel subgroup in a growing superfamily of proteins regulating diverse physiological processes in bacterial pathogens.

    PubMed

    Thomson, N R; Cox, A; Bycroft, B W; Stewart, G S; Williams, P; Salmond, G P

    1997-11-01

    The enteric bacterium Serratia marcescens is an opportunistic human pathogen. The strain ATCC39006 makes the red pigment, prodigiosin (Pig), and the beta-lactam antibiotic carbapenem (Car). Mutants were isolated that were concomitantly defective for Pig and Car production. These mutants were found to have a mutation in the rap gene (Regulation of Antibiotic and Pigment). Sequence analysis of the rap gene revealed a predicted protein product showing strong homology to SlyA, originally thought to be a haemolytic virulence determinant in Salmonella typhimurium. Homologues of rap were detected in several bacterial genera, including Salmonella, Yersinia, Enterobacter, and species of the plant pathogen, Erwinia. The Erwinia hoeEr (homologue of rap) and the Yersinia horYe genes were also found to be very similar to rap and slyA. Marker exchange mutagenesis of horEr revealed that it encoded a regulatory protein controlling the production of antibiotic and exoenzyme virulence determinants in the phytopathogen, Erwinia carotovora subspecies carotovora. We have shown that these new homologues of SlyA form a highly conserved subgroup of a growing superfamily of bacterial regulatory proteins controlling diverse physiological processes in human, animal and plant pathogens.

  16. Allowable residual contamination levels of radionuclides in soil from pathway analysis

    SciTech Connect

    Nyquist, J.E.; Baes, C.F. III

    1987-01-01

    The uncertainty regarding radionuclide distributions among Remedial Action Program (RAP) sites and long-term decommissioning and closure options for these sites requires a flexible approach capable of handling different levels of contamination, dose limits, and closure scenarios. We identified a commercially available pathway analysis model, DECOM, which had been used previously in support of remedial activities involving contaminated soil at the Savannah River Plant. The DECOM computer code, which estimates concentrations of radionuclides uniformly distributed in soil that correspond to an annual effective dose equivalent, is written in BASIC and runs on an IBM PC or compatible microcomputer. We obtained the latest version of DECOM and modified it to make it more user friendly and applicable to the Oak Ridge National Laboratory (ORNL) RAP. Some modifications involved changes in default parameters or changes in models based on approaches used by the EPA in regulating remedial actions for hazardous substances. We created a version of DECOM as a LOTUS spreadsheet, using the same models as the BASIC version of DECOM. We discuss the specific modeling approaches taken, the regulatory framework that guided our efforts, the strengths and limitations of each approach, and areas for improvement. We also demonstrate how the LOTUS version of DECOM can be applied to specific problems that may be encountered during ORNL RAP activities. 18 refs., 2 figs., 3 tabs.

  17. Scientific Ability and Creativity

    ERIC Educational Resources Information Center

    Heller, Kurt A.

    2007-01-01

    Following an introductory definition of "scientific ability and creativity", product-oriented, personality and social psychological approaches to studying scientific ability are examined with reference to competence and performance. Studies in the psychometric versus cognitive psychological paradigms are dealt with in more detail. These two…

  18. Mixed Ability Teaching.

    ERIC Educational Resources Information Center

    Skov, Poul

    1986-01-01

    As a basis for taking a position on the future school structure in grades 8-10 in Denmark, an extensive study was carried out on mixed ability teaching (teaching in heterogeneous classes) on these grade levels. Results showed that mixed ability teaching gave at least as good results as teaching in differentiated classes. (Author/LMO)

  19. Fairness and Ability Grouping.

    ERIC Educational Resources Information Center

    Strike, Kenneth A.

    1983-01-01

    A recent controversy regarding ability grouping is that it is often perceived as a means whereby racial or class bias can be subtly transformed into mechanisms of discrimination which exhibit the appearance of fairness and objectivity. This article addresses the question of fairness in ability grouping. (CJB)

  20. The purification of a Rap1 GTPase-activating protein from bovine brain cytosol.

    PubMed

    Nice, E C; Fabri, L; Hammacher, A; Holden, J; Simpson, R J; Burgess, A W

    1992-01-25

    Two GTPase-activating proteins (GAPs) have been detected in extracts from bovine brain: GAP-1, which is specific for the activation of ras GTPases, and GAP-3, which is specific for the activation of the rap1 GTPases. We present a strategy for the purification to homogeneity of a cytosolic form of GAP-3 from bovine brain. The 100,000 x g supernatant from homogenized brains was chromatographed sequentially on DEAE Fast Flow, green H-E4BD Sepharose, Bio-Gel A1.5, hydroxyapatite, and phenyl-Sepharose prior to high resolution separation on Mono Q HR 5/5, phenyl-Superose HR 5/5, Mono Q PC 1.6/5, and Superose 12 PC 3.2/30. This procedure resulted in an approximately 18,000-fold purification, yielding 50 micrograms of GAP-3 from 1.6 kg of tissue. Purified cytosolic GAP-3 migrated as a single band of apparent Mr 55,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, on gel filtration cytosolic GAP-3 chromatographed as a dimer with an apparent Mr 92,000. Purified GAP-3 does not activate ras or rho GTPases and possesses no intrinsic GTPase activity. Amino acid sequence data indicated a proline-rich N terminus. The amino acid sequences of peptides generated by Staphylococcus aureus V8 digestion of reduced and pyridine-ethylated GAP-3 showed no similarity to the predicted primary structure of GAP-1 or any other proteins in the nucleic acid or protein data bases. By comparison with the data of Rubinfeld et al. (Rubinfeld, B., Munemitsu, S., Clark, R., Conroy, L., Watt, K., Crosier, W.J., McCormick, F., and Polakis, P. (1991) Cell 65, 1033-1042), it appears that the membrane-associated (Mr 85,000-95,000) and cytosolic forms of GAP-3 are derived from equivalent, or closely related, genes. PMID:1309786

  1. Evidence for a role of rap1 protein in the regulation of human platelet Ca2+ fluxes.

    PubMed Central

    Corvazier, E; Enouf, J; Papp, B; de Gunzburg, J; Tavitian, A; Levy-Toledano, S

    1992-01-01

    The relationship between the 22-24 kDa cyclic AMP (cAMP)-dependent phosphoprotein previously described as being involved in the regulation of human platelet membrane Ca2+ transport and a GTP-binding protein of low molecular mass (ras-like protein) was investigated. After isolation of plasma membranes and intracellular membranes, it was found that guanosine 5'-[gamma-thio]triphosphate (GTP[S]) bound to plasma membrane proteins ranging in molecular mass from 22 to 29 kDa, but not to intracellular membranes. The major GTP-binding protein appeared as a 24 kDa protein under reduced conditions and a 22 kDa protein under non-reduced conditions. A similar membrane location and electrophoretic mobility were found for both the cAMP phosphoprotein and the protein recognized by a specific anti-rap1 antibody. The identity between the cAMP phosphoprotein and the rap1 GTP-binding protein was further examined by studying the functional effect of GTP on plasma membrane Ca2+ transport. A maximal GTP[S] concentration of 40 microM was found to: (1) inhibit to the same degree (40%) both Ca(2+)-ATPase activity and the Ca2+ transport function mediated by the Ca(2+)-ATPase; (2) inhibit the phosphorylation of the 22-24 kDa protein by the catalytic subunit of the cAMP-dependent protein kinase (C.Sub.); and (3) abolish the stimulation of Ca2+ uptake induced by C.Sub. It is concluded that the platelet cAMP phosphoprotein is indeed the rap1 GTP-binding protein, and that it regulates plasma membrane Ca2+ transport, thus providing evidence for a new role of a ras-related protein. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:1310590

  2. RapMap: a rapid, sensitive and accurate tool for mapping RNA-seq reads to transcriptomes

    PubMed Central

    Srivastava, Avi; Sarkar, Hirak; Gupta, Nitish; Patro, Rob

    2016-01-01

    Motivation: The alignment of sequencing reads to a transcriptome is a common and important step in many RNA-seq analysis tasks. When aligning RNA-seq reads directly to a transcriptome (as is common in the de novo setting or when a trusted reference annotation is available), care must be taken to report the potentially large number of multi-mapping locations per read. This can pose a substantial computational burden for existing aligners, and can considerably slow downstream analysis. Results: We introduce a novel concept, quasi-mapping, and an efficient algorithm implementing this approach for mapping sequencing reads to a transcriptome. By attempting only to report the potential loci of origin of a sequencing read, and not the base-to-base alignment by which it derives from the reference, RapMap—our tool implementing quasi-mapping—is capable of mapping sequencing reads to a target transcriptome substantially faster than existing alignment tools. The algorithm we use to implement quasi-mapping uses several efficient data structures and takes advantage of the special structure of shared sequence prevalent in transcriptomes to rapidly provide highly-accurate mapping information. We demonstrate how quasi-mapping can be successfully applied to the problems of transcript-level quantification from RNA-seq reads and the clustering of contigs from de novo assembled transcriptomes into biologically meaningful groups. Availability and implementation: RapMap is implemented in C ++11 and is available as open-source software, under GPL v3, at https://github.com/COMBINE-lab/RapMap. Contact: rob.patro@cs.stonybrook.edu Supplementary information: Supplementary data are available at Bioinformatics online. PMID:27307617

  3. Measuring creative imagery abilities

    PubMed Central

    Jankowska, Dorota M.; Karwowski, Maciej

    2015-01-01

    Over the decades, creativity and imagination research developed in parallel, but they surprisingly rarely intersected. This paper introduces a new theoretical model of creative visual imagination, which bridges creativity and imagination research, as well as presents a new psychometric instrument, called the Test of Creative Imagery Abilities (TCIA), developed to measure creative imagery abilities understood in accordance with this model. Creative imagination is understood as constituted by three interrelated components: vividness (the ability to create images characterized by a high level of complexity and detail), originality (the ability to produce unique imagery), and transformativeness (the ability to control imagery). TCIA enables valid and reliable measurement of these three groups of abilities, yielding the general score of imagery abilities and at the same time making profile analysis possible. We present the results of nine studies on a total sample of more than 1700 participants, showing the factor structure of TCIA using confirmatory factor analysis, as well as provide data confirming this instrument's validity and reliability. The availability of TCIA for interested researchers may result in new insights and possibilities of integrating the fields of creativity and imagination science. PMID:26539140

  4. Can reading rate acceleration improve error monitoring and cognitive abilities underlying reading in adolescents with reading difficulties and in typical readers?

    PubMed

    Horowitz-Kraus, Tzipi; Breznitz, Zvia

    2014-01-28

    Dyslexia is characterized by slow, inaccurate reading and by deficits in executive functions. The deficit in reading is exemplified by impaired error monitoring, which can be specifically shown through neuroimaging, in changes in Error-/Correct-related negativities (ERN/CRN). The current study aimed to investigate whether a reading intervention program (Reading Acceleration Program, or RAP) could improve overall reading, as well as error monitoring and other cognitive abilities underlying reading, in adolescents with reading difficulties. Participants with reading difficulties and typical readers were trained with the RAP for 8 weeks. Their reading and error monitoring were characterized both behaviorally and electrophysiologically through a lexical decision task. Behaviorally, the reading training improved "contextual reading speed" and decreased reading errors in both groups. Improvements were also seen in speed of processing, memory and visual screening. Electrophysiologically, ERN increased in both groups following training, but the increase was significantly greater in the participants with reading difficulties. Furthermore, an association between the improvement in reading speed and the change in difference between ERN and CRN amplitudes following training was seen in participants with reading difficulties. These results indicate that improving deficits in error monitoring and speed of processing are possible underlying mechanisms of the RAP intervention. We suggest that ERN is a good candidate for use as a measurement in evaluating the effect of reading training in typical and disabled readers.

  5. Genetic analysis of innate immunity in Behcet’s disease identifies an association with IL-37 and IL-18RAP

    PubMed Central

    Tan, Handan; Deng, Bolin; Yu, Hongsong; Yang, Yi; Ding, Lin; Zhang, Qi; Qin, Jieying; Kijlstra, Aize; Chen, Rui; Yang, Peizeng

    2016-01-01

    Interleukin-1 (IL-1) and the IL-1 receptor (IL-1R) family play an important role in the pathogenesis of inflammatory diseases. This study aimed to investigate the association between single nucleotide polymorphisms (SNP) of IL-1 and IL-1R family genes with Vogt-Koyanagi-Harada (VKH) and Behcet’s disease (BD) in Han Chinese. The case-control study was divided into two stages and included 419 VKH cases, 1063 BD cases and 1872 healthy controls. The MassARRAY platform (Sequenom), iPLEX Gold Assay and TaqMan SNP assays were used to score genotypes of 24 SNPs. The expression of IL-37 and IL-18Rap was measured by ELISA and real-time PCR in genotyped healthy individuals. A significantly lower frequency of the AG genotype, and a higher frequency of the GG genotype and G allele of IL-37/rs3811047 were observed in BD as compared to controls. AA genotype and A allele frequency of IL-18RAP/rs2058660 was significantly decreased in BD as compared to controls. Functional studies performed in healthy controls showed that rs3811047 AG genotype carriers had a higher IL-37 gene expression in peripheral blood mononuclear cells (PBMCs) than GG carriers. GG carriers showed a higher cytokine expression as compared to AG carriers. No association was detected between the tested SNPs and VKH. PMID:27775096

  6. Ground and flight calibration assessment of HiRAP accelerometer data from missions STS-35 and STS-40

    NASA Technical Reports Server (NTRS)

    Blanchard, Robert C.; Larman, Kevin T.; Moats, Christina D.

    1992-01-01

    A method of removing non-aerodynamic signals and calibrating the High Resolution Accelerometer Package (HiRAP) flight data was developed and is discussed for Shuttle Orbiter missions STS-35 and STS-40. These two mission data sets were analyzed using ground (dynamic) calibration data and flight calibrations using a flight calibration technique that was developed and refined over the HiRAP operational lifetime. This technique evolved early in the flight program, since it was recognized that ground calibration factors are insufficient to determine absolute low acceleration levels. The application of flight calibration factors to the data sets from these missions produced calibrated acceleration levels within an accuracy of less than plus or minus 1.5 micro-g of zero during a time in the flight when the acceleration level was known to be less than 1 micro-g. This analysis further confirms the theory that flight calibrations are required in order to obtain the absolute measurement of low-frequency, low-acceleration flight signals.

  7. Critical function of RA-GEF-2/Rapgef6, a guanine nucleotide exchange factor for Rap1, in mouse spermatogenesis.

    PubMed

    Okada, Keisuke; Miyake, Hideaki; Yamaguchi, Kohei; Chiba, Koji; Maeta, Kazuhiro; Bilasy, Shymaa E; Edamatsu, Hironori; Kataoka, Tohru; Fujisawa, Masato

    2014-02-28

    Small GTPase Rap1 has been implicated in the proper differentiation of testicular germ cells. In the present study, we investigated the functional significance of RA-GEF-2/Rapgef6, a guanine nucleotide exchange factor for Rap1, in testicular differentiation using mice lacking RA-GEF-2. RA-GEF-2 was expressed predominantly on the luminal side of the seminiferous tubules in wild-type mice. No significant differences were observed in the body weights or hormonal parameters of RA-GEF-2(-)(/)(-) and wild-type mice. However, the testes of RA-GEF-2(-)(/)(-) male mice were significantly smaller than those of wild-type mice and were markedly atrophied as well as hypospermatogenic. The concentration and motility of epididymal sperm were also markedly reduced and frequently had an abnormal shape. The pregnancy rate and number of fetuses were markedly lower in wild-type females after they mated with RA-GEF-2(-)(/)(-) males than with wild-type males, which demonstrated the male infertility phenotype of RA-GEF-2(-)(/)(-) mice. Furthermore, a significant reduction and alteration were observed in the expression level and cell junctional localization of N-cadherin, respectively, in RA-GEF-2(-)(/)(-) testes, which may, at least in part, account for the defects in testicular differentiation and spermatogenesis in these mice. PMID:24491570

  8. RAP80-directed tuning of BRCA1 homologous recombination function at ionizing radiation-induced nuclear foci

    PubMed Central

    Hu, Yiduo; Scully, Ralph; Sobhian, Bijan; Xie, Anyong; Shestakova, Elena; Livingston, David M.

    2011-01-01

    In response to DNA double-strand breaks (DSBs), BRCA1 forms biochemically distinct complexes with certain other DNA damage response proteins. These structures, some of which are required for homologous recombination (HR)-type DSB repair, concentrate at distinct nuclear foci that demarcate sites of genome breakage. Polyubiquitin binding by one of these structures, the RAP80/BRCA1 complex, is required for efficient BRCA1 focal recruitment, but the relationship of this process to the execution of HR has been unclear. We found that this complex actively suppresses otherwise exaggerated, BRCA1-driven HR. By controlling the kinetics by which other BRCA1-interacting proteins that promote HR concentrate together with BRCA1 in nuclear foci, RAP80/BRCA1 complexes suppress excessive DSB end processing, HR-type DSB repair, and overt chromosomal instability. Since chromosomal instability emerges when BRCA1 HR function is either unbridled or absent, active tuning of BRCA1 activity, executed in nuclear foci, is important to genome integrity maintenance. PMID:21406551

  9. Is Pluto a planet? Student powered video rap ';battle' over tiny Pluto's embattled planetary standing

    NASA Astrophysics Data System (ADS)

    Beisser, K.; Cruikshank, D. P.; McFadden, T.

    2013-12-01

    Is Pluto a planet? Some creative low income Bay-area middle-schoolers put a musical spin on this hot science debate with a video rap ';battle' over tiny Pluto's embattled planetary standing. The students' timing was perfect, with NASA's New Horizons mission set to conduct the first reconnaissance of Pluto and its moons in July 2015. Pluto - the last of the nine original planets to be explored by spacecraft - has been the subject of scientific study and speculation since Clyde Tombaugh discovered it in 1930, orbiting the Sun far beyond Neptune. Produced by the students and a very creative educator, the video features students 'battling' back and forth over the idea of Pluto being a planet. The group collaborated with actual space scientists to gather information and shot their video before a 'green screen' that was eventually filled with animations and visuals supplied by the New Horizons mission team. The video debuted at the Pluto Science Conference in Maryland in July 2013 - to a rousing response from researchers in attendance. The video marks a nontraditional approach to the ongoing 'great planet debate' while educating viewers on a recently discovered region of the solar system. By the 1990s, researchers had learned that Pluto possessed multiple exotic ices on its surface, a complex atmosphere and seasonal cycles, and a large moon (Charon) that likely resulted from a giant impact on Pluto itself. It also became clear that Pluto was no misfit among the planets - as had long been thought - but the largest and brightest body in a newly discovered 'third zone' of our planetary system called the Kuiper Belt. More recent observations have revealed that Pluto has a rich system of satellites - five known moons - and a surface that changes over time. Scientists even speculate that Pluto may possess an internal ocean. For these and other reasons, the 2003 Planetary Decadal Survey ranked a Pluto/Kuiper Belt mission as the highest priority mission for NASA's newly created

  10. Human abilities: emotional intelligence.

    PubMed

    Mayer, John D; Roberts, Richard D; Barsade, Sigal G

    2008-01-01

    Emotional intelligence (EI) involves the ability to carry out accurate reasoning about emotions and the ability to use emotions and emotional knowledge to enhance thought. We discuss the origins of the EI concept, define EI, and describe the scope of the field today. We review three approaches taken to date from both a theoretical and methodological perspective. We find that Specific-Ability and Integrative-Model approaches adequately conceptualize and measure EI. Pivotal in this review are those studies that address the relation between EI measures and meaningful criteria including social outcomes, performance, and psychological and physical well-being. The Discussion section is followed by a list of summary points and recommended issues for future research. PMID:17937602

  11. Who's Playin' Whom? Overwhelming Influence of Hip-Hop Culture, Rap Music on Historically Black College and University (HBCU) Campuses Concerns Students; Faculty

    ERIC Educational Resources Information Center

    Stewart, Pearl

    2004-01-01

    In December 2000, Dr. Thomas Earl Midgette had harsh words for the hip-hop movement that was sweeping his campus. When he was interviewed for an article in "Black Issues" titled "The Miseducation of Hip-Hop," Midgette didn't hold back: "You see students walking on campus reciting rap lyrics when they should be reciting something they'll need to…

  12. Sequential sentinel SNP Regional Association Plots (SSS-RAP): an approach for testing independence of SNP association signals using meta-analysis data.

    PubMed

    Zheng, Jie; Gaunt, Tom R; Day, Ian N M

    2013-01-01

    Genome-Wide Association Studies (GWAS) frequently incorporate meta-analysis within their framework. However, conditional analysis of individual-level data, which is an established approach for fine mapping of causal sites, is often precluded where only group-level summary data are available for analysis. Here, we present a numerical and graphical approach, "sequential sentinel SNP regional association plot" (SSS-RAP), which estimates regression coefficients (beta) with their standard errors using the meta-analysis summary results directly. Under an additive model, typical for genes with small effect, the effect for a sentinel SNP can be transformed to the predicted effect for a possibly dependent SNP through a 2×2 2-SNP haplotypes table. The approach assumes Hardy-Weinberg equilibrium for test SNPs. SSS-RAP is available as a Web-tool (http://apps.biocompute.org.uk/sssrap/sssrap.cgi). To develop and illustrate SSS-RAP we analyzed lipid and ECG traits data from the British Women's Heart and Health Study (BWHHS), evaluated a meta-analysis for ECG trait and presented several simulations. We compared results with existing approaches such as model selection methods and conditional analysis. Generally findings were consistent. SSS-RAP represents a tool for testing independence of SNP association signals using meta-analysis data, and is also a convenient approach based on biological principles for fine mapping in group level summary data.

  13. Priming Ability Emotional Intelligence

    ERIC Educational Resources Information Center

    Schutte, Nicola S.; Malouff, John M.

    2012-01-01

    Two studies examined whether priming self-schemas relating to successful emotional competency results in better emotional intelligence performance. In the first study participants were randomly assigned to a successful emotional competency self-schema prime condition or a control condition and then completed an ability measure of emotional…

  14. Higher Level Thinking Abilities.

    ERIC Educational Resources Information Center

    Mills, Barbara, Ed.

    This report describes two systems designed to improve teaching competencies and to develop higher level thinking abilities, and presents the evaluation design, statistical results, and a brief history of the major events which occurred during development. The McCollum-Davis Model is designed to develop understanding of and skill in relating a…

  15. A Specific Calculating Ability.

    ERIC Educational Resources Information Center

    Anderson, Mike; O'Connor, Neil; Hermelin, Beate

    1998-01-01

    Studied the calculating ability used by a low IQ savant to identify prime numbers in two experiments comparing him to control subjects, one involving reaction time and the other involving inspection time. Concludes that this individual uses a complex computational algorithm to identify primes and discusses the apparent contradiction of his low IQ.…

  16. E3B1, a human homologue of the mouse gene product Abi-1, sensitizes activation of Rap1 in response to epidermal growth factor

    SciTech Connect

    Jenei, Veronika; Andersson, Tommy; Jakus, Judit; Dib, Karim . E-mail: k.dib@qub.ac.uk

    2005-11-01

    E3B1, a human homologue of the mouse gene product Abi-1, has been implicated in growth-factor-mediated regulation of the small GTPases p21{sup Ras} and Rac. E3b1 is a regulator of Rac because it can form a complex with Sos-1 and eps8, and such a Sos-1-e3B1-eps8 complex serves as a guanine nucleotide exchange factor for Rac. In the present study, we found that overexpression of e3B1 in NIH3T3/EGFR cells sensitized EGF-induced activation of Rac1, whereas it had no impact on EGF-induced activation of p21{sup Ras}. Remarkably, we found that EGF-induced activation of the p21{sup Ras}-related GTPase Rap1 was also sensitized in NIH3T3/EGFR-e3B1 cells. Thus, in NIH3T3/EGFR-e3B1 cells, maximal EGF-induced activation of Rap1 occurs with a dose of EGF much lower than in NIH3T3/EGFR cells. We also report that overexpression of e3B1 in NIH3T3/EGFR cells renders EGF-induced activation of Rap1 completely dependent on Src tyrosine kinases but not on c-Abl. However, EGF-induced tyrosine phosphorylation of the Rap GEF C3G occurred regardless of whether e3B1 was overexpressed or not, and this did not involve Src tyrosine kinases. Accordingly, we propose that overexpression of e3B1 in NIH3T3/EGFR cells leads to mobilization of Src tyrosine kinases that participate in EGF-induced activation of Rap1 and inhibition of cell proliferation.

  17. The best marker for guiding the clinical management of patients with raised intracranial pressure-the RAP index or the mean pulse amplitude?

    PubMed

    Hall, Allan; O'Kane, Roddy

    2016-10-01

    Raised intracranial pressure is a common problem in a variety of neurosurgical conditions including traumatic brain injury, hydrocephalus and intracranial haemorrhage. The clinical management of these patients is guided by a variety of haemodynamic, biochemical and clinical factors. However to date there is no single parameter that is used to guide clinical management of patients with raised intracranial pressure (ICP). However, the role of ICP indices, specifically the mean pulse amplitude (AMP) and RAP index [correlation coefficient (R) between AMP amplitude (A) and mean ICP pressure (P); index of compensatory reserve], as an indicator of true ICP has been investigated. Whilst the RAP index has been used both as a descriptor of neurological deterioration in TBI patients and as a way of characterising the compensatory reserve in hydrocephalus, more recent studies have highlighted the limitation of the RAP index due to the influence that baseline effect errors have on the mean ICP, which is used in the calculation of the RAP index. These studies have suggested that the ICP mean pulse amplitude may be a more accurate marker of true intracranial pressure due to the fact that it is uninfluenced by the mean ICP and, therefore, the AMP may be a more reliable marker than the RAP index for guiding the clinical management of patients with raised ICP. Although further investigation needs to be undertaken in order to fully assess the role of ICP indices in guiding the clinical management of patients with raised ICP, the studies undertaken to date provide an insight into the potential role of ICP indices to treat raised ICP proactively rather than reactively and therefore help prevent or minimise secondary brain injury.

  18. The best marker for guiding the clinical management of patients with raised intracranial pressure-the RAP index or the mean pulse amplitude?

    PubMed

    Hall, Allan; O'Kane, Roddy

    2016-10-01

    Raised intracranial pressure is a common problem in a variety of neurosurgical conditions including traumatic brain injury, hydrocephalus and intracranial haemorrhage. The clinical management of these patients is guided by a variety of haemodynamic, biochemical and clinical factors. However to date there is no single parameter that is used to guide clinical management of patients with raised intracranial pressure (ICP). However, the role of ICP indices, specifically the mean pulse amplitude (AMP) and RAP index [correlation coefficient (R) between AMP amplitude (A) and mean ICP pressure (P); index of compensatory reserve], as an indicator of true ICP has been investigated. Whilst the RAP index has been used both as a descriptor of neurological deterioration in TBI patients and as a way of characterising the compensatory reserve in hydrocephalus, more recent studies have highlighted the limitation of the RAP index due to the influence that baseline effect errors have on the mean ICP, which is used in the calculation of the RAP index. These studies have suggested that the ICP mean pulse amplitude may be a more accurate marker of true intracranial pressure due to the fact that it is uninfluenced by the mean ICP and, therefore, the AMP may be a more reliable marker than the RAP index for guiding the clinical management of patients with raised ICP. Although further investigation needs to be undertaken in order to fully assess the role of ICP indices in guiding the clinical management of patients with raised ICP, the studies undertaken to date provide an insight into the potential role of ICP indices to treat raised ICP proactively rather than reactively and therefore help prevent or minimise secondary brain injury. PMID:27567609

  19. Cortex Mori Radicis Extract induces neurite outgrowth in PC12 cells activating ERK signaling pathway via inhibiting Ca2+ influx

    PubMed Central

    Yin, Nina; Hong, Xiaoping; Han, Yongming; Duan, Yanjun; Zhang, Yanhong; Chen, Zebin

    2015-01-01

    Cortex Mori Radicis is a traditional Chinese herbal medicine which has a long history of use for the treatment of headaches, cough, edema and diabetes. However, its function and mode of action within nervous system remain largely unclear. In the present study, we have attempted to determine the effects of Cortex Mori Radicis Extract (CMRE) on neuronal differentiation. Here, we reported that CMRE induces the neurite outgrowth in pheochromocytoma PC12 cells and primary cortical neuron. Following the generation of neurite outgrowth, extracellular Ca2+ influx was inhibited and intracellular Ca2+ decreased. In addition, CMRE induced the extracellular signal-regulated kinase 1/2 (ERK1/2) activation and also stimulated the Rap1-GTP expression, which is closely linked to neuritogenesis. Moreover, the neurite outgrowth induced by CMRE was antagonized to a marked degree by suppressing activation of p-ERK1/2 with the specific ERK1/2 inhibitor (PD98059), suggesting the involvement of Rap1-GTP and ERK1/2 in CMRE-induced neurite outgrowth. Taken together, these results demonstrate that CMRE induces neurite outgrowth of PC12 cells through Rap1-ERK signaling pathway via inhibiting Ca2+ influx, and provide a novel insight into the manner in which CMRE participates in neuritogenesis. PMID:26131075

  20. Nutrient Sensing Mechanisms and Pathways

    PubMed Central

    Efeyan, Alejo; Comb, William C.; Sabatini, David M.

    2015-01-01

    PREFACE The ability to sense and respond to fluctuations in environmental nutrient levels is a requisite for life. Nutrient scarcity is a selective pressure that has shaped the evolution of most cellular processes. Different pathways that detect intracellular and extracellular levels of sugars, amino acids and lipids, and surrogate metabolites, are then integrated and coordinated at the organismal level via hormonal signals. During food abundance, nutrient sensing pathways engage anabolism and storage, and scarcity triggers homeostatic mechanisms, like the mobilization of internal stores through mechanisms such as autophagy. Nutrient sensing pathways are commonly deregulated in human metabolic diseases. PMID:25592535

  1. Exome sequencing identifies recessive CDK5RAP2 variants in patients with isolated agenesis of corpus callosum.

    PubMed

    Jouan, Loubna; Ouled Amar Bencheikh, Bouchra; Daoud, Hussein; Dionne-Laporte, Alexandre; Dobrzeniecka, Sylvia; Spiegelman, Dan; Rochefort, Daniel; Hince, Pascale; Szuto, Anna; Lassonde, Maryse; Barbelanne, Marine; Tsang, William Y; Dion, Patrick A; Théoret, Hugo; Rouleau, Guy A

    2016-04-01

    Agenesis of the corpus callosum (ACC) is a common brain malformation which can be observed either as an isolated condition or as part of numerous congenital syndromes. Therefore, cognitive and neurological involvements in patients with ACC are variable, from mild linguistic and behavioral impairments to more severe neurological deficits. To date, the underlying genetic causes of isolated ACC remains elusive and causative genes have yet to be identified. We performed exome sequencing on three acallosal siblings from the same non-consanguineous family and identified compound heterozygous variants, p.[Gly94Arg];[Asn1232Ser], in the protein encoded by the CDK5RAP2 gene, also known as MCPH3, a gene previously reported to cause autosomal recessive primary microcephaly. Our findings suggest a novel role for this gene in the pathogenesis of isolated ACC. PMID:26197979

  2. Hidden Worries

    ERIC Educational Resources Information Center

    Reclaiming Children and Youth, 2013

    2013-01-01

    Sometimes children are stressed about seemingly small events that escalate into problem behavior. In this article, an insightful teacher discusses restoration of emotional balance by mobilizing positive support from both school and family using A Response Ability Pathways (RAP) intervention. The RAP techniques and how they can be used are…

  3. General English Ability, Specific Purpose English Ability, and Computer Skills.

    ERIC Educational Resources Information Center

    Prapphal, Kanchana

    2003-01-01

    Aims to answer the following research questions: (1) Are general English ability and specific purpose English ability related to computer skills? and (2) Is general English ability transferable to specific purpose English ability? Subjects were third year science students enrolled in an English for academic purposes course. (Author/VWL)

  4. Cdc42 and k-Ras Control Endothelial Tubulogenesis through Apical Membrane and Cytoskeletal Polarization: Novel Stimulatory Roles for GTPase Effectors, the Small GTPases, Rac2 and Rap1b, and Inhibitory Influence of Arhgap31 and Rasa1

    PubMed Central

    Norden, Pieter R.; Kim, Dae Joong; Barry, David M.; Cleaver, Ondine B.; Davis, George E.

    2016-01-01

    A critical and understudied property of endothelial cells is their ability to form lumens and tube networks. Although considerable information has been obtained concerning these issues, including the role of Cdc42 and Rac1 and their effectors such as Pak2, Pak4, Par6b, and co-regulators such as integrins, MT1-MMP and Par3; many key questions remain that are necessary to elucidate molecular and signaling requirements for this fundamental process. In this work, we identify new small GTPase regulators of EC tubulogenesis including k-Ras, Rac2 and Rap1b that act in conjunction with Cdc42 as well as the key downstream effectors, IQGAP1, MRCKβ, beta-Pix, GIT1, and Rasip1 (which can assemble into multiprotein complexes with key regulators including α2β1 integrin and MT1-MMP). In addition, we identify the negative regulators, Arhgap31 (by inactivating Cdc42 and Rac) and Rasa1 (by inactivating k-Ras) and the positive regulator, Arhgap29 (by inactivating RhoA) which play a major functional role during the EC tubulogenic process. Human EC siRNA suppression or mouse knockout of Rasip1 leads to identical phenotypes where ECs form extensive cord networks, but cannot generate lumens or tubes. Essential roles for these molecules during EC tubulogenesis include; i) establishment of asymmetric EC cytoskeletal polarization (subapical distribution of acetylated tubulin and basal membrane distribution of F-actin); and ii) directed membrane trafficking of pinocytic vacuoles or other intracellular vesicles along acetylated tubulin tracks to the developing apical membrane surface. Cdc42 co-localizes subapically with acetylated tubulin, while Rac1 and k-Ras strongly label vacuole/ vesicle membranes which accumulate and fuse together in a polarized, perinuclear manner. We observe polarized apical membrane and subapical accumulation of key GTPases and effectors regulating EC lumen formation including Cdc42, Rac1, Rac2, k-Ras, Rap1b, activated c-Raf and Rasip1 to control EC tube network

  5. Cdc42 and k-Ras Control Endothelial Tubulogenesis through Apical Membrane and Cytoskeletal Polarization: Novel Stimulatory Roles for GTPase Effectors, the Small GTPases, Rac2 and Rap1b, and Inhibitory Influence of Arhgap31 and Rasa1.

    PubMed

    Norden, Pieter R; Kim, Dae Joong; Barry, David M; Cleaver, Ondine B; Davis, George E

    2016-01-01

    A critical and understudied property of endothelial cells is their ability to form lumens and tube networks. Although considerable information has been obtained concerning these issues, including the role of Cdc42 and Rac1 and their effectors such as Pak2, Pak4, Par6b, and co-regulators such as integrins, MT1-MMP and Par3; many key questions remain that are necessary to elucidate molecular and signaling requirements for this fundamental process. In this work, we identify new small GTPase regulators of EC tubulogenesis including k-Ras, Rac2 and Rap1b that act in conjunction with Cdc42 as well as the key downstream effectors, IQGAP1, MRCKβ, beta-Pix, GIT1, and Rasip1 (which can assemble into multiprotein complexes with key regulators including α2β1 integrin and MT1-MMP). In addition, we identify the negative regulators, Arhgap31 (by inactivating Cdc42 and Rac) and Rasa1 (by inactivating k-Ras) and the positive regulator, Arhgap29 (by inactivating RhoA) which play a major functional role during the EC tubulogenic process. Human EC siRNA suppression or mouse knockout of Rasip1 leads to identical phenotypes where ECs form extensive cord networks, but cannot generate lumens or tubes. Essential roles for these molecules during EC tubulogenesis include; i) establishment of asymmetric EC cytoskeletal polarization (subapical distribution of acetylated tubulin and basal membrane distribution of F-actin); and ii) directed membrane trafficking of pinocytic vacuoles or other intracellular vesicles along acetylated tubulin tracks to the developing apical membrane surface. Cdc42 co-localizes subapically with acetylated tubulin, while Rac1 and k-Ras strongly label vacuole/ vesicle membranes which accumulate and fuse together in a polarized, perinuclear manner. We observe polarized apical membrane and subapical accumulation of key GTPases and effectors regulating EC lumen formation including Cdc42, Rac1, Rac2, k-Ras, Rap1b, activated c-Raf and Rasip1 to control EC tube network

  6. Design of High-Affinity Stapled Peptides To Target the Repressor Activator Protein 1 (RAP1)/Telomeric Repeat-Binding Factor 2 (TRF2) Protein-Protein Interaction in the Shelterin Complex.

    PubMed

    Ran, Xu; Liu, Liu; Yang, Chao-Yie; Lu, Jianfeng; Chen, Yong; Lei, Ming; Wang, Shaomeng

    2016-01-14

    Shelterin, a six-protein complex, plays a fundamental role in protecting both the length and the stability of telomeres. Repressor activator protein 1 (RAP1) and telomeric repeat-binding factor 2 (TRF2) are two subunits in shelterin that interact with each other. Small-molecule inhibitors that block the RAP1/TRF2 protein-protein interaction can disrupt the structure of shelterin and may be employed as pharmacological tools to investigate the biology of shelterin. On the basis of the cocrystal structure of RAP1/TRF2 complex, we have developed first-in-class triazole-stapled peptides that block the protein-protein interaction between RAP1 and TRF2. Our most potent stapled peptide binds to RAP1 protein with a Ki value of 7 nM and is >100 times more potent than the corresponding wild-type TRF2 peptide. On the basis of our high-affinity peptides, we have developed and optimized a competitive, fluorescence polarization (FP) assay for accurate and rapid determination of the binding affinities of our designed compounds and this assay may also assist in the discovery of non-peptide, small-molecule inhibitors capable of blocking the RAP1/TRF2 protein-protein interaction.

  7. Biological activity and redistribution of nucleolar proteins of two different cell lines treated with cis-dichloro-1,2-propylenediamine-N,N,N',N'-tetraacetato ruthenium (III) (RAP).

    PubMed

    Delmani, Fatima Azzahra; Torreblanca, José; Moreno, Javier; García-Herdugo, Gregorio; Vilaplana, Rosario; González-Víltchez, Francisco

    2014-06-01

    The interaction of a newly synthesized antitumor complex cis-dichloro-1,2-propylenediamine-N,N,N',N'-tetraacetato ruthenium (III) (RAP) with DNA was investigated in vitro through a number of techniques including comet assay, immunoprecipitation, and immunolocalization of certain nucleolar proteins (the upstream binding factor (UBF) and fibrillarin) involved in DNA transcription, rRNA processing, and ribosomal assembly. The results showed that RAP binds to the DNA of two cell lines (H4 and Hs-683) causing a delay in cell proliferation rate leading to a number of cellular modifications. These modifications include DNA-damage assessed by the single cell gel electrophoresis method (comet assay) and variation in the expression of nucleolar proteins; UBF was more abundant in RAP treated cells, this was explained by the high affinity of this protein to DNA modified by RAP. On the other hand, fibrillarin was found in less quantities in RAP treated cells which was explained by a de-regulation of the ribosomal machinery caused by RAP.

  8. Impaired musical ability in people with schizophrenia

    PubMed Central

    Hatada, Sanae; Sawada, Ken; Akamatsu, Masanori; Doi, Erina; Minese, Masayoshi; Yamashita, Motoshi; Thornton, Allen E.; Honer, William G.; Inoue, Shimpei

    2014-01-01

    Background Assessment of the musical ability of people with schizophrenia has attracted little interest despite the diverse and substantive findings of impairments in sound perception and processing and the therapeutic effect of music in people with the illness. The present study investigated the musical ability of people with schizophrenia and the association with psychiatric symptoms and cognition. Methods We recruited patients with chronic schizophrenia and healthy controls for participation in our study. To measure musical ability and cognitive function, we used the Montreal Battery of Evaluation of Amusia (MBEA) and the Brief Assessment of Cognition in Schizophrenia (BACS). We carried out a mediation analysis to investigate a possible pathway to a deficit in musical ability. Results We enrolled 50 patients and 58 controls in the study. The MBEA global score in patients with schizophrenia was significantly lower than that in controls (p < 0.001), and was strongly associated with both the composite cognitive function score (r = 0.645, p < 0.001) and the negative symptom score (r = −0.504, p < 0.001). Further analyses revealed direct and indirect effects of negative symptoms on musical ability. The indirect effects were mediated through cognitive impairment. Limitations The relatively small sample size did not permit full evaluation of the possible effects of age, sex, education, medication and cultural influences on the results. Conclusion Examining the associations between musical deficits, negative symptoms and cognitive imapirment in patients with schizophrenia may identify shared biological mechanisms. PMID:24119791

  9. Rap1 GTPase Inhibits Tumor Necrosis Factor-α-Induced Choroidal Endothelial Migration via NADPH Oxidase- and NF-κB-Dependent Activation of Rac1.

    PubMed

    Wang, Haibo; Fotheringham, Lori; Wittchen, Erika S; Hartnett, M Elizabeth

    2015-12-01

    Macrophage-derived tumor necrosis factor (TNF)-α has been found in choroidal neovascularization (CNV) surgically removed from patients with age-related macular degeneration. However, the role of TNF-α in CNV development remains unclear. In a murine laser-induced CNV model, compared with un-lasered controls, TNF-α mRNA was increased in retinal pigment epithelial and choroidal tissue, and TNF-α colocalized with lectin-stained migrating choroidal endothelial cells (CECs). Inhibition of TNF-α with a neutralizing antibody reduced CNV volume and reactive oxygen species (ROS) level around CNV. In CECs, pretreatment with the antioxidant apocynin or knockdown of p22phox, a subunit of NADPH oxidase, inhibited TNF-α-induced ROS generation. Apocynin reduced TNF-α-induced NF-κB and Rac1 activation, and inhibited TNF-α-induced CEC migration. TNF-α-induced Rac1 activation and CEC migration were inhibited by NF-κB inhibitor Bay11-7082. Overexpression of Rap1a prevented TNF-α-induced ROS generation and reduced NF-κB and Rac1 activation. Activation of Rap1 by 8-(4-chlorophenylthio)adenosine-2'-O-Me-cAMP prevented TNF-α-induced CEC migration and reduced laser-induced CNV volume, ROS generation, and activation of NF-κB and Rac1. These findings provide evidence that active Rap1a inhibits TNF-α-induced CEC migration by inhibiting NADPH oxidase-dependent NF-κB and Rac1 activation and suggests that Rap1a de-escalates CNV development by interfering with ROS-dependent signaling in several steps of the pathogenic process. PMID:26476350

  10. Specialized Rap1p/Gcr1p Transcriptional Activation through Gcr1p DNA Contacts Requires Gcr2p, as Does Hyperphosphorylation of Gcr1p

    PubMed Central

    Zeng, X.; Deminoff, S. J.; Santangelo, G. M.

    1997-01-01

    The multifunctional regulatory factor Rap1p of Saccharomyces cerevisiae accomplishes one of its tasks, transcriptional activation, by complexing with Gcr1p. An unusual feature of this heteromeric complex is its apparent capacity to contact simultaneously two adjacent DNA elements (UAS(RPG) and the CT box, bound specifically by Rap1p and Gcr1p, respectively). The complex can activate transcription through isolated UAS(RPG) but not CT elements. In promoters that contain both DNA signals its activity is enhanced, provided the helical spacing between the two elements is appropriate; this suggests that at least transient DNA loop formation is involved. We show here that this CT box-dependent augmentation of Rap1p/Gcr1p activation requires the presence of a third protein Gcr2p; the Gcr2(-) growth defect appears to result from a genome-wide loss of the CT box effect. Interestingly, a hyperphosphorylated form of Gcr1p disappears in Δgcr2 cells but reappears if they harbor a doubly point-mutated GCR1 allele that bypasses the Gcr2(-) growth defect. Gcr2p therefore appears to induce a conformation change in Gcr1p and/or stimulate its hyperphosphorylation; one or both of these effects can be mimicked in the absence of GCR2 by mutation of GCR1. This improved view of Rap1p/Gcr1p/Gcr2p function reveals a new aspect of eukaryotic gene regulation: modification of an upstream activator, accompanied by at least transient DNA loop formation, mediates its improved capacity to activate transcription. PMID:9335588

  11. Chromosomal localization of human genes for the LDL receptor family member glycoprotein 330 (LRP2) and its associated protein RAP (LRPAP1)

    SciTech Connect

    Korenberg, J.R.; Chen, X.N.; Argraves, K.M.

    1994-07-01

    Glycoprotein 330 (gp330) is a member of a family of receptors with structural similarities to the low-density lipoprotein receptor. Gp330 is expressed by a number of specialized epithelia, including renal proximal tubules, where it can mediate endocytosis of ligands such as complexes of urokinase and the serpin, plasminogen activator inhibitor-1. Gp330 has also been shown to bind in vitro to lipoprotein lipase and apolipoprotein E-enriched {beta}VLDL, suggesting a role for this receptor in lipoprotein metabolism. The 39-kDa protein, referred to as receptor associated protein (RAP), binds to and copurifies with gp330 and antagonizes the ligand binding activity of gp330. In this paper, the authors report the use of homology-PCR cloning to isolate cDNAs encoding human gp330. Using gp330 cDNA and previously isolated human RAP cDNA probes, they performed fluorescence in situ hybridization to map the human chromosomal location of the genes for these proteins. The gene for gp330 was mapped at a single site on the long arm of human chromosome 2 on the border of bands 2q24-q31. The gene for RAP was mapped to the short arm of human chromosome 4 at position 4q16.3, which is in the region of the chromosomal deletion causing Wolf-Hirschhorn syndrome. The assignment of chromosomal map positions for gp330 and RAP genes will aid in the evaluation of their potential roles in human diseases such as Wolf-Hirschhorn syndrome and disorders of lipoprotein metabolism, such as atherosclerosis. 38 refs., 3 figs., 1 tab.

  12. Spatially selective hormonal control of RAP2.6L and ANAC071 transcription factors involved in tissue reunion in Arabidopsis.

    PubMed

    Asahina, Masashi; Azuma, Katsuya; Pitaksaringkarn, Weerasak; Yamazaki, Takashi; Mitsuda, Nobutaka; Ohme-Takagi, Masaru; Yamaguchi, Shinjiro; Kamiya, Yuji; Okada, Kiyotaka; Nishimura, Takeshi; Koshiba, Tomokazu; Yokota, Takao; Kamada, Hiroshi; Satoh, Shinobu

    2011-09-20

    When grafting or wounding disconnects stem tissues, new tissues are generated to restore the lost connection. In this study, the molecular mechanism of such healing was elucidated in injured stems of Arabidopsis. Soon after the inflorescence stems were incised, the pith cells started to divide. This process was strongly inhibited by the elimination of cauline leaves, shoot apices, or lateral buds that reduced the indole-3-acetic acid supply. Microarray and quantitative RT-PCR analyses revealed that genes related to cell division, phytohormones, and transcription factors were expressed because of incision. Among them, two plant-specific transcription factor genes, ANAC071 and RAP2.6L, were abundantly expressed. ANAC071 was expressed at 1-3 d after cutting exclusively in the upper region of the cut gap, with concomitant accumulation of indole-3-acetic acid. In contrast, RAP2.6L was expressed at 1 d after cutting exclusively in the lower region, with concomitant deprivation of indole-3-acetic acid. The expression of ANAC071 and RAP2.6L were also promoted by ethylene and jasmonic acid, respectively. In transformants suppressing the function of RAP2.6L or ANAC071, the division of pith cells was inhibited. Furthermore, the ethylene signaling-defective ein2 mutant showed incomplete healing. Hence, plant-specific transcription factors differentially expressed around the cut position were essential for tissue reunion of Arabidopsis wounded flowering stems and were under opposite control by polar-transported auxin, with modification by the ethylene and jasmonic acid wound-inducible hormones.

  13. Development and evaluation of RapTAT: a machine learning system for concept mapping of phrases from medical narratives.

    PubMed

    Gobbel, Glenn T; Reeves, Ruth; Jayaramaraja, Shrimalini; Giuse, Dario; Speroff, Theodore; Brown, Steven H; Elkin, Peter L; Matheny, Michael E

    2014-04-01

    Rapid, automated determination of the mapping of free text phrases to pre-defined concepts could assist in the annotation of clinical notes and increase the speed of natural language processing systems. The aim of this study was to design and evaluate a token-order-specific naïve Bayes-based machine learning system (RapTAT) to predict associations between phrases and concepts. Performance was assessed using a reference standard generated from 2860 VA discharge summaries containing 567,520 phrases that had been mapped to 12,056 distinct Systematized Nomenclature of Medicine - Clinical Terms (SNOMED CT) concepts by the MCVS natural language processing system. It was also assessed on the manually annotated, 2010 i2b2 challenge data. Performance was established with regard to precision, recall, and F-measure for each of the concepts within the VA documents using bootstrapping. Within that corpus, concepts identified by MCVS were broadly distributed throughout SNOMED CT, and the token-order-specific language model achieved better performance based on precision, recall, and F-measure (0.95±0.15, 0.96±0.16, and 0.95±0.16, respectively; mean±SD) than the bag-of-words based, naïve Bayes model (0.64±0.45, 0.61±0.46, and 0.60±0.45, respectively) that has previously been used for concept mapping. Precision, recall, and F-measure on the i2b2 test set were 92.9%, 85.9%, and 89.2% respectively, using the token-order-specific model. RapTAT required just 7.2ms to map all phrases within a single discharge summary, and mapping rate did not decrease as the number of processed documents increased. The high performance attained by the tool in terms of both accuracy and speed was encouraging, and the mapping rate should be sufficient to support near-real-time, interactive annotation of medical narratives. These results demonstrate the feasibility of rapidly and accurately mapping phrases to a wide range of medical concepts based on a token-order-specific naïve Bayes model and

  14. Development and evaluation of RapTAT: a machine learning system for concept mapping of phrases from medical narratives.

    PubMed

    Gobbel, Glenn T; Reeves, Ruth; Jayaramaraja, Shrimalini; Giuse, Dario; Speroff, Theodore; Brown, Steven H; Elkin, Peter L; Matheny, Michael E

    2014-04-01

    Rapid, automated determination of the mapping of free text phrases to pre-defined concepts could assist in the annotation of clinical notes and increase the speed of natural language processing systems. The aim of this study was to design and evaluate a token-order-specific naïve Bayes-based machine learning system (RapTAT) to predict associations between phrases and concepts. Performance was assessed using a reference standard generated from 2860 VA discharge summaries containing 567,520 phrases that had been mapped to 12,056 distinct Systematized Nomenclature of Medicine - Clinical Terms (SNOMED CT) concepts by the MCVS natural language processing system. It was also assessed on the manually annotated, 2010 i2b2 challenge data. Performance was established with regard to precision, recall, and F-measure for each of the concepts within the VA documents using bootstrapping. Within that corpus, concepts identified by MCVS were broadly distributed throughout SNOMED CT, and the token-order-specific language model achieved better performance based on precision, recall, and F-measure (0.95±0.15, 0.96±0.16, and 0.95±0.16, respectively; mean±SD) than the bag-of-words based, naïve Bayes model (0.64±0.45, 0.61±0.46, and 0.60±0.45, respectively) that has previously been used for concept mapping. Precision, recall, and F-measure on the i2b2 test set were 92.9%, 85.9%, and 89.2% respectively, using the token-order-specific model. RapTAT required just 7.2ms to map all phrases within a single discharge summary, and mapping rate did not decrease as the number of processed documents increased. The high performance attained by the tool in terms of both accuracy and speed was encouraging, and the mapping rate should be sufficient to support near-real-time, interactive annotation of medical narratives. These results demonstrate the feasibility of rapidly and accurately mapping phrases to a wide range of medical concepts based on a token-order-specific naïve Bayes model and

  15. Different Dimensions of Spatial Ability.

    ERIC Educational Resources Information Center

    Eliot, John; Hauptman, Anna

    1981-01-01

    Indicates that spatial ability describes a variety of different behaviors and briefly reviews efforts to define intelligence factors and identify processes involved in solving tasks requiring spatial ability. (DS)

  16. Generation of a Potent Low Density Lipoprotein Receptor-related Protein 1 (LRP1) Antagonist by Engineering a Stable Form of the Receptor-associated Protein (RAP) D3 Domain.

    PubMed

    Prasad, Joni M; Migliorini, Mary; Galisteo, Rebeca; Strickland, Dudley K

    2015-07-10

    The low density lipoprotein receptor-related protein 1 (LRP1) is a member of the low density lipoprotein receptor family and plays important roles in a number of physiological and pathological processes. Expression of LRP1 requires the receptor-associated protein (RAP), a molecular chaperone that binds LRP1 and other low density lipoprotein receptor family members in the endoplasmic reticulum and traffics with them to the Golgi where the acidic environment causes its dissociation. Exogenously added RAP is a potent LRP1 antagonist and binds to LRP1 on the cell surface, preventing ligands from binding. Following endocytosis, RAP dissociates in the acidic endosome, allowing LRP1 to recycle back to the cell surface. The acid-induced dissociation of RAP is mediated by its D3 domain, a relatively unstable three-helical bundle that denatures at pH <6.2 due to protonation of key histidine residues on helices 2 and 3. To develop an LRP1 inhibitor that does not dissociate at low pH, we introduced a disulfide bond between the second and third helices in the RAP D3 domain. By combining this disulfide bond with elimination of key histidine residues, we generated a stable RAP molecule that is resistant to both pH- and heat-induced denaturation. This molecule bound to LRP1 with high affinity at both neutral and acidic pH and proved to be a potent inhibitor of LRP1 function both in vitro and in vivo, suggesting that our stable RAP molecule may be useful in multiple pathological settings where LRP1 blockade has been shown to be effective.

  17. An Association Study of Interleukin 18 Receptor Genes (IL18R1 and IL18RAP) in Lumbar Disc Degeneration

    PubMed Central

    Omair, Ahmad; Lie, Benedicte Alexandra; Reikeras, Olav; Brox, Jens Ivar

    2012-01-01

    Objectives: To examine association of candidate genetic variants in structural, inflammatory, matrix modifying, vitamin D receptor genes and variants associated with osteoarthritis, with surgical candidates and surgical patients with lumbar disc degeneration (LDD), in light of their previously reported susceptibility for LDD. Methods: Genotyping of 146 Norwegian LDD patients and 188 Norwegian controls was performed for 20 single-nucleotide polymorphisms (SNPs) from collagen, aggrecan, interleukin, VDR, MMP3 and COX2 genes and 7 SNPs from osteoarthritic genes. Results: The neighboring genes IL18R1 and IL18RAP polymorphisms (rs2287037 and rs1420100), showed a statistically non-significant risk for developing LDD (OR 1.36 [95 % CI 0.99 – 1.87]; p=0.06 and OR 1.33 [95 % CI 0.98-1.81]; p=0.07). Homozygosity of these risk alleles was associated with LDD (p=0.023 and p=0.027). The non-risk alleles at these SNPs were situated on a haplotype negatively associated with LDD (p=0.008). Carriage of at least one non-risk allele at both loci also reduces the risk of developing LDD (OR 0.51 [95 % CI 0.33-0.80]; p=0.003). Conclusion: Our findings support the polygenic nature of LDD and suggest that variation in interleukin 18 receptor genes could affect the risk of severe LDD and associated low back pain. PMID:22550553

  18. Environmental performance and mechanical analysis of concrete containing recycled asphalt pavement (RAP) and waste precast concrete as aggregate.

    PubMed

    Erdem, Savaş; Blankson, Marva Angela

    2014-01-15

    The overall objective of this research project was to investigate the feasibility of incorporating 100% recycled aggregates, either waste precast concrete or waste asphalt planning, as replacements for virgin aggregates in structural concrete and to determine the mechanical and environmental performance of concrete containing these aggregates. Four different types of concrete mixtures were designed with the same total water cement ratio (w/c=0.74) either by using natural aggregate as reference or by totally replacing the natural aggregate with recycled material. Ground granulated blast furnace slag (GGBS) was used as a mineral addition (35%) in all mixtures. The test results showed that it is possible to obtain satisfactory performance for strength characteristics of concrete containing recycled aggregates, if these aggregates are sourced from old precast concrete. However, from the perspective of the mechanical properties, the test results indicated that concrete with RAP aggregate cannot be used for structural applications. In terms of leaching, the results also showed that the environmental behaviour of the recycled aggregate concrete is similar to that of the natural aggregate concrete. PMID:24316812

  19. Environmental performance and mechanical analysis of concrete containing recycled asphalt pavement (RAP) and waste precast concrete as aggregate.

    PubMed

    Erdem, Savaş; Blankson, Marva Angela

    2014-01-15

    The overall objective of this research project was to investigate the feasibility of incorporating 100% recycled aggregates, either waste precast concrete or waste asphalt planning, as replacements for virgin aggregates in structural concrete and to determine the mechanical and environmental performance of concrete containing these aggregates. Four different types of concrete mixtures were designed with the same total water cement ratio (w/c=0.74) either by using natural aggregate as reference or by totally replacing the natural aggregate with recycled material. Ground granulated blast furnace slag (GGBS) was used as a mineral addition (35%) in all mixtures. The test results showed that it is possible to obtain satisfactory performance for strength characteristics of concrete containing recycled aggregates, if these aggregates are sourced from old precast concrete. However, from the perspective of the mechanical properties, the test results indicated that concrete with RAP aggregate cannot be used for structural applications. In terms of leaching, the results also showed that the environmental behaviour of the recycled aggregate concrete is similar to that of the natural aggregate concrete.

  20. TRPC3 amplifies B-cell receptor-induced ERK signalling via protein kinase D-dependent Rap1 activation.

    PubMed

    Numaga-Tomita, Takuro; Nishida, Motohiro; Putney, James W; Mori, Yasuo

    2016-01-15

    Sustained activation of extracellular-signal-regulated kinase (ERK) has an important role in the decision regarding the cell fate of B-lymphocytes. Recently, we demonstrated that the diacylglycerol-activated non-selective cation channel canonical transient receptor potential 3 (TRPC3) is required for the sustained ERK activation induced by the B-cell receptor. However, the signalling mechanism underlying TRPC3-mediated ERK activation remains elusive. In the present study, we have shown that TRPC3 mediates Ca(2+) influx to sustain activation of protein kinase D (PKD) in a protein kinase C-dependent manner in DT40 B-lymphocytes. The later phase of ERK activation depends on the small G-protein Rap1, known as a downstream target of PKD, whereas the earlier phase of ERK activation depends on the Ras protein. It is of interest that sustained ERK phosphorylation is required for the full induction of the immediate early gene Egr-1 (early growth response 1). These results suggest that TRPC3 reorganizes the BCR signalling complex by switching the subtype of small G-proteins to sustain ERK activation in B-lymphocytes.