Science.gov

Sample records for ability pathways rap

  1. Functional interaction between p21rap1A and components of the budding pathway in Saccharomyces cerevisiae.

    PubMed Central

    McCabe, P C; Haubruck, H; Polakis, P; McCormick, F; Innis, M A

    1992-01-01

    The rap1A gene encodes a 21-kDa, ras-related GTP-binding protein (p21rap1A) of unknown function. A close structural homolog of p21rap1A (65% identity in the amino-terminal two-thirds) is the RSR1 gene product (Rsr1p) of Saccharomyces cerevisiae. Although Rsr1p is not essential for growth, its presence is required for nonrandom selection of bud sites. To assess the similarity of these proteins at the functional level, wild-type and mutant forms of p21rap1A were tested for complementation of activities known to be fulfilled by Rsr1p. Expression of p21rap1A, like multicopy expression of RSR1, suppressed the conditional lethality of a temperature-sensitive cdc24 mutation. Point mutations predicted to affect the localization of p21rap1A or its ability to cycle between GDP and GTP-bound states disrupted suppression of cdc24ts, while other mutations in the 61-65 loop region improved suppression. Expression of p21rap1A could not, however, suppress the random budding phenotype of rsr1 cells. p21rap1A also apparently interfered with the normal activity of Rsrlp, causing random budding in diploid wild-type cells, suggesting an inability of p21rap1A to interact appropriately with Rsr1p regulatory proteins. Consistent with this hypothesis, we found an Rsr1p-specific GTPase-activating protein (GAP) activity in yeast membranes which was not active toward p21rap1A, indicating that p21rap1A may be predominantly GTP bound in yeast cells. Coexpression of human Rap1-specific GAP suppressed the random budding due to expression of p21rap1A or its derivatives, including Rap1AVal-12. Although Rap1-specific GAP stimulated the GTPase of Rsr1p in vitro, it did not dominantly interfere with Rsr1p function in vivo. A chimera consisting of Rap1A1-165::Rsr1p166-272 did not exhibit normal Rsr1p function in the budding pathway. These results indicated that p21rap1A and Rsr1p share at least partial functional homology, which may have implications for p21rap1A function in mammalian cells. Images

  2. 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.…

  3. 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…

  4. PRL-3 activates NF-κB signaling pathway by interacting with RAP1.

    PubMed

    Lian, Shenyi; Meng, Lin; Liu, Caiyun; Xing, Xiaofang; Song, Qian; Dong, Bin; Han, Yong; Yang, Yongyong; Peng, Lirong; Qu, Like; Shou, Chengchao

    2013-01-04

    Phosphatase of regenerating liver (PRL-3) promotes cancer metastasis through enhanced cell motility and invasiveness, however its role in tumorigenesis remains unclear. Herein, we reported that PRL-3 interacts with telomere-related protein RAP1. PRL-3 promotes the cytosolic localization of RAP1, which is counteracted by silencing of PRL-3. Immunohistochemical staining of colon cancer tissue array (n=170) revealed that high level of PRL-3 associates with cytosolic localization of RAP1 (p=0.01). Microarray analysis showed that PRL-3 regulates expression of diverse genes and enhances phosphorylation of p65 subunit of NF-κB in a RAP1-dependent manner. Furthermore, PRL-3 transcriptionally activates RAP1 expression, which is counteracted by ablating p65. Therefore, our results demonstrate PRL-3 as a novel regulator of NF-κB signaling pathway through RAP1.

  5. 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

  6. CDK5RAP2 interaction with components of the Hippo signaling pathway may play a role in primary microcephaly.

    PubMed

    Sukumaran, Salil K; Stumpf, Maria; Salamon, Sarah; Ahmad, Ilyas; Bhattacharya, Kurchi; Fischer, Sarah; Müller, Rolf; Altmüller, Janine; Budde, Birgit; Thiele, Holger; Tariq, Muhammad; Malik, Naveed Altaf; Nürnberg, Peter; Baig, Shahid Mahmood; Hussain, Muhammad Sajid; Noegel, Angelika A

    2017-04-01

    Autosomal recessive primary microcephaly (MCPH) is characterized by a substantial reduction in brain size but with normal architecture. It is often linked to mutations in genes coding for centrosomal proteins; however, their role in brain size regulation is not completely understood. By combining homozygosity mapping and whole-exome sequencing in an MCPH family from Pakistan, we identified a novel mutation (XM_011518861.1; c.4114C > T) in CDK5RAP2, the gene associated with primary microcephaly-3 (MCPH3), leading to a premature stop codon (p.Arg1372*). CDK5RAP2 is a component of the pericentriolar material important for the microtubule-organizing function of the centrosome. Patient-derived primary fibroblasts had strongly decreased CDK5RAP2 amounts, showed centrosomal and nuclear abnormalities and exhibited changes in cell size and migration. We further identified an interaction of CDK5RAP2 with the Hippo pathway components MST1 kinase and the transcriptional regulator TAZ. This finding potentially provides a mechanism through which the Hippo pathway with its roles in the regulation of centrosome number is linked to the centrosome. In the patient fibroblasts, we observed higher levels of TAZ and YAP. However, common target genes of the Hippo pathway were downregulated as compared to the control with the exception of BIRC5 (Survivin), which was significantly upregulated. We propose that the centrosomal deficiencies and the altered cellular properties in the patient fibroblasts can also result from the observed changes in the Hippo pathway components which could thus be relevant for MCPH and play a role in brain size regulation and development.

  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.

  8. cAMP signaling increases histone deacetylase 8 expression via the Epac2–Rap1A–Akt pathway in H1299 lung cancer cells

    PubMed Central

    Park, Ji-Yeon; Juhnn, Yong-Sung

    2017-01-01

    This study was performed to investigate the signaling pathway that mediates cyclic AMP (cAMP)-induced inhibition of histone deacetylase 8 (HDAC8) degradation, and the effect and underlying mechanisms of the resulting increase in HDAC8 expression on cisplatin-induced apoptosis in lung cancer cells. cAMP signaling increased HDAC8 expression via a protein kinase A (PKA)-independent pathway in H1299 non-small cell lung cancer cells. However, treatment with a selective activator of an exchange protein that was activated by cAMP (Epac) increased HDAC8 expression, and Epac2 inhibition abolished the isoproterenol (ISO)-induced increase in HDAC8 expression. ISO and the Epac activator activated Rap1, and Rap1A activation increased HDAC8 expression; moreover, inhibition of Rap1A with a dominant negative Rap1A or by shRNA-mediated knockdown abolished the ISO-induced increase in HDAC8 expression. Activation of cAMP signaling and Rap1A decreased the activating phosphorylation of Akt. Akt inhibition with a pharmacological inhibitor or expression of a dominant negative Akt inhibited the MKK4/JNK pathway and increased HDAC8 expression. The Akt inhibitor-induced increase in HDAC8 expression was abolished by pretreatment with proteasomal or lysosomal inhibitors. The ISO treatment increased cisplatin-induced apoptosis, which was abolished by HDAC8 knockdown. Exogenous HDAC8 expression increased cisplatin-induced apoptosis and decreased TIPRL expression, and the knockdown of TIPRL increased the apoptosis of cisplatin-treated cells. The ISO treatment decreased cisplatin-induced transcription of the TIPRL gene in a HDAC8-dependent manner. In conclusion, the Epac–Rap1–Akt pathway mediates cAMP signaling-induced inhibition of JNK-dependent HDAC8 degradation, and the resulting HDAC8 increase augments cisplatin-induced apoptosis by repressing TIPRL expression in H1299 lung cancer cells. PMID:28232663

  9. cAMP signaling increases histone deacetylase 8 expression via the Epac2-Rap1A-Akt pathway in H1299 lung cancer cells.

    PubMed

    Park, Ji-Yeon; Juhnn, Yong-Sung

    2017-02-24

    This study was performed to investigate the signaling pathway that mediates cyclic AMP (cAMP)-induced inhibition of histone deacetylase 8 (HDAC8) degradation, and the effect and underlying mechanisms of the resulting increase in HDAC8 expression on cisplatin-induced apoptosis in lung cancer cells. cAMP signaling increased HDAC8 expression via a protein kinase A (PKA)-independent pathway in H1299 non-small cell lung cancer cells. However, treatment with a selective activator of an exchange protein that was activated by cAMP (Epac) increased HDAC8 expression, and Epac2 inhibition abolished the isoproterenol (ISO)-induced increase in HDAC8 expression. ISO and the Epac activator activated Rap1, and Rap1A activation increased HDAC8 expression; moreover, inhibition of Rap1A with a dominant negative Rap1A or by shRNA-mediated knockdown abolished the ISO-induced increase in HDAC8 expression. Activation of cAMP signaling and Rap1A decreased the activating phosphorylation of Akt. Akt inhibition with a pharmacological inhibitor or expression of a dominant negative Akt inhibited the MKK4/JNK pathway and increased HDAC8 expression. The Akt inhibitor-induced increase in HDAC8 expression was abolished by pretreatment with proteasomal or lysosomal inhibitors. The ISO treatment increased cisplatin-induced apoptosis, which was abolished by HDAC8 knockdown. Exogenous HDAC8 expression increased cisplatin-induced apoptosis and decreased TIPRL expression, and the knockdown of TIPRL increased the apoptosis of cisplatin-treated cells. The ISO treatment decreased cisplatin-induced transcription of the TIPRL gene in a HDAC8-dependent manner. In conclusion, the Epac-Rap1-Akt pathway mediates cAMP signaling-induced inhibition of JNK-dependent HDAC8 degradation, and the resulting HDAC8 increase augments cisplatin-induced apoptosis by repressing TIPRL expression in H1299 lung cancer cells.

  10. 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

  11. 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.

  12. 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…

  13. 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…

  14. Optimal expression condition of recombinant RAP.

    PubMed

    Zhang, Jie; Zhang, Hong; Bi, Hao; Liu, Zhiguo; Guo, Jianli; Qu, Shen

    2007-02-01

    In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

  15. 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.

  16. [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.

  17. Functions for the cAMP/Epac/Rap1 Signaling Pathway in Low-Dose Endothelial Monocyte-Activating Polypeptide-II-Induced Opening of Blood-Tumor Barrier.

    PubMed

    Li, Zhen; Liu, Xiao-Bai; Liu, Yun-Hui; Xue, Yi-Xue; Wang, Ping; Liu, Li-Bo; Yao, Yi-Long; Ma, Jun

    2015-09-01

    Previous studies have demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) induces blood-tumor barrier (BTB) hyperpermeability via both paracellular and transcellular pathways. In a recent study, we revealed that cAMP/PKA-dependent and cAMP/PKA-independent signaling pathways are both involved in EMAP-II-induced BTB hyperpermeability. The present study further investigated the exact mechanisms through which the cAMP/PKA-independent signaling pathway affects EMAP-II-induced BTB hyperpermeability. In an in vitro BTB model, low-dose EMAP-II (0.05 nM) induced a significant decrease in Rap1 activity in RBMECs. Pretreatment with forskolin to elevate intracellular cAMP concentration completely blocked EMAP-II-induced Rap1 inactivation. Epac/Rap1 activation by 8-pCPT-2'-O-Me-cAMP significantly prevented EMAP-II-induced activation of RhoA/ROCK. Furthermore, 8-pCPT-2'-O-Me-cAMP pretreatment significantly inhibited EMAP-II-induced decreases in TEER and increases in HRP flux. Pretreatment also significantly prevented EMAP-II-induced changes in MLC phosphorylation, actin cytoskeleton arrangement, and expression and distribution of ZO-1 in RBMECs. This study demonstrates that the cAMP/Epac/Rap1 signaling cascade is a crucial pathway in EMAP-II-induced BTB hyperpermeability.

  18. The Fluids RAP

    NASA Astrophysics Data System (ADS)

    Nedyalkov, Ivaylo

    2016-11-01

    After fifteen years of experience in rap, and ten in fluid mechanics, "I am coming here with high-Reynolds-number stamina; I can beat these rap folks whose flows are... laminar." The rap relates fluid flows to rap flows. The fluid concepts presented in the song have varying complexity and the listeners/viewers will be encouraged to read the explanations on a site dedicated to the rap. The music video will provide an opportunity to share high-quality fluid visualizations with a general audience. This talk will present the rap lyrics, the vision for the video, and the strategy for outreach. Suggestions and comments will be welcomed.

  19. Unfolding of the RAP-D3 helical bundle facilitates dissociation of RAP-receptor complexes.

    PubMed

    Estrada, Kristine; Fisher, Carl; Blacklow, Stephen C

    2008-02-12

    The receptor-associated protein (RAP) functions as an escort protein for receptors of the low-density lipoprotein receptor (LDLR) family by preventing premature intracellular binding of ligands and assisting with delivery of mature receptors to the cell surface. The modulation of affinity by pH is believed to play an important role in the escort function of RAP, because RAP binds tightly to proteins of the LDLR family at near-neutral pH early in the secretory pathway where its high affinity precludes premature binding of ligands but then dissociates from bound receptors at the lower pH of the Golgi compartment. The third domain of RAP (RAP-D3), which forms a three-helix bundle, is sufficient to reconstitute the escort activity. Here, we test the hypothesis that low-pH induced unfolding of the RAP-D3 helical bundle facilitates dissociation of RAP-receptor complexes. First, variants of RAP-D3 resistant to low pH-induced unfolding were constructed by replacing interior histidine residues with phenylalanines. In contrast to native RAP-D3, which exhibits an unfolding pKa of 6.3 and a Tm of 42 degrees C, the most hyperstable variant of RAP-D3, in which four histidine residues are replaced with phenylalanine, has an unfolding pKa of 4.8, and a Tm of 58 degrees C. The phenylalanine substitutions in RAP-D3 confer increased stability to pH-induced dissociation of complexes formed between RAP-D3 and a two-repeat fragment of the LDLR (LA3-4). When introduced into full-length RAP, the four mutations that confer hyperstability on RAP-D3 interfere with transport of endogenous LRP-1 to the cell surface in a dominant negative fashion under conditions where expression of normal RAP has no effect on LRP-1 transport. Our studies support a model in which low pH-dependent unfolding of RAP-D3 facilitates dissociation of RAP from the LA repeats of LDLR family proteins in the mildly acidic pH of the Golgi.

  20. Positive Use of Rap Music in the Classroom.

    ERIC Educational Resources Information Center

    Anderson, Edward

    As an extension of African-Americans' rich language and musical heritage and abilities, rap music has some value in the educational setting. Rap music started as a dance fad beginning in the mid-1970s among Blacks and Hispanics in New York's outer boroughs. It is another generational brand of Black language and musical usage and an extension of…

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. Dynamin2 controls Rap1 activation and integrin clustering in human T lymphocyte adhesion

    PubMed Central

    Eppler, Felix J.

    2017-01-01

    Leukocyte trafficking is crucial to facilitate efficient immune responses. Here, we report that the large GTPase dynamin2, which is generally considered to have a key role in endocytosis and membrane remodeling, is an essential regulator of integrin-dependent human T lymphocyte adhesion and migration. Chemical inhibition or knockdown of dynamin2 expression significantly reduced integrin-dependent T cell adhesion in vitro. This phenotype was not observed when T cells were treated with various chemical inhibitors which abrogate endocytosis or actin polymerization. We furthermore detected dynamin2 in signaling complexes and propose that it controls T cell adhesion via FAK/Pyk2- and RapGEF1-mediated Rap1 activation. In addition, the dynamin2 inhibitor-induced reduction of lymphocyte adhesion can be rescued by Rap1a overexpression. We demonstrate that the dynamin2 effect on T cell adhesion does not involve integrin affinity regulation but instead relies on its ability to modulate integrin valency. Taken together, we suggest a previously unidentified role of dynamin2 in the regulation of integrin-mediated lymphocyte adhesion via a Rap1 signaling pathway. PMID:28273099

  6. TFIIA and the transactivator Rap1 cooperate to commit TFIID for transcription initiation.

    PubMed

    Papai, Gabor; Tripathi, Manish K; Ruhlmann, Christine; Layer, Justin H; Weil, P Anthony; Schultz, Patrick

    2010-06-17

    Transcription of eukaryotic messenger RNA (mRNA) encoding genes by RNA polymerase II (Pol II) is triggered by the binding of transactivating proteins to enhancer DNA, which stimulates the recruitment of general transcription factors (TFIIA, B, D, E, F, H) and Pol II on the cis-linked promoter, leading to pre-initiation complex formation and transcription. In TFIID-dependent activation pathways, this general transcription factor containing TATA-box-binding protein is first recruited on the promoter through interaction with activators and cooperates with TFIIA to form a committed pre-initiation complex. However, neither the mechanisms by which activation signals are communicated between these factors nor the structural organization of the activated pre-initiation complex are known. Here we used cryo-electron microscopy to determine the architecture of nucleoprotein complexes composed of TFIID, TFIIA, the transcriptional activator Rap1 and yeast enhancer-promoter DNA. These structures revealed the mode of binding of Rap1 and TFIIA to TFIID, as well as a reorganization of TFIIA induced by its interaction with Rap1. We propose that this change in position increases the exposure of TATA-box-binding protein within TFIID, consequently enhancing its ability to interact with the promoter. A large Rap1-dependent DNA loop forms between the activator-binding site and the proximal promoter region. This loop is topologically locked by a TFIIA-Rap1 protein bridge that folds over the DNA. These results highlight the role of TFIIA in transcriptional activation, define a molecular mechanism for enhancer-promoter communication and provide structural insights into the pathways of intramolecular communication that convey transcription activation signals through the TFIID complex.

  7. Rap Music in the Classroom?

    ERIC Educational Resources Information Center

    Anderson, Edward

    1993-01-01

    Discusses the background of rap music, its definition, its themes and messages, and rap as a blend of language and music. Offers ideas for its use in the classroom as a way to motivate and instruct students. (SR)

  8. 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…

  9. 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,…

  10. The structure and conformational switching of Rap1B.

    PubMed

    Noguchi, Hiroki; Ikegami, Takahisa; Nagadoi, Aritaka; Kamatari, Yuji O; Park, Sam-Yong; Tame, Jeremy R H; Unzai, Satoru

    2015-06-19

    Rap1B is a small GTPase involved in the regulation of numerous cellular processes including synaptic plasticity, one of the bases of memory. Like other members of the Ras family, the active GTP-bound form of Rap1B can bind to a large number of effector proteins and so transmit signals to downstream components of the signaling pathways. The structure of Rap1B bound only to a nucleotide has yet to be solved, but might help reveal an inactive conformation that can be stabilized by a small molecule drug. Unlike other Ras family proteins such as H-Ras and Rap2A, Rap1B crystallizes in an intermediate state when bound to a non-hydrolyzable GTP analog. Comparison with H-Ras and Rap2A reveals conservative mutations relative to Rap1B, distant from the bound nucleotide, which control how readily the protein may adopt the fully activated form in the presence of GTP. High resolution crystallographic structures of mutant proteins show how these changes may influence the hydrogen bonding patterns of the key switch residues.

  11. Impaired TIP60-mediated H4K16 acetylation accounts for the aberrant chromatin accumulation of 53BP1 and RAP80 in Fanconi anemia pathway-deficient cells.

    PubMed

    Renaud, Emilie; Barascu, Aurelia; Rosselli, Filippo

    2016-01-29

    To rescue collapsed replication forks cells utilize homologous recombination (HR)-mediated mechanisms to avoid the induction of gross chromosomal abnormalities that would be generated by non-homologous end joining (NHEJ). Using DNA interstrand crosslinks as a replication barrier, we investigated how the Fanconi anemia (FA) pathway promotes HR at stalled replication forks. FA pathway inactivation results in Fanconi anemia, which is associated with a predisposition to cancer. FANCD2 monoubiquitination and assembly in subnuclear foci appear to be involved in TIP60 relocalization to the chromatin to acetylates histone H4K16 and prevents the binding of 53BP1 to its docking site, H4K20Me2. Thus, FA pathway loss-of-function results in accumulation of 53BP1, RIF1 and RAP80 at damaged chromatin, which impair DNA resection at stalled replication fork-associated DNA breaks and impede HR. Consequently, DNA repair in FA cells proceeds through the NHEJ pathway, which is likely responsible for the accumulation of chromosome abnormalities. We demonstrate that the inhibition of NHEJ or deacetylase activity rescue HR in FA cells.

  12. Rap Music and Choral Education.

    ERIC Educational Resources Information Center

    Bitz, Michael

    1998-01-01

    Suggests choral teachers use rap music to promote student interest and to teach music basics, such as rhythm, pitch, harmony, and timbre. Maintains that students can write the arrangements allowing them to gain experience in notating. Identifies selected recordings and offers an example of how to use rap music. (CMK)

  13. 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…

  14. Activation of G protein-coupled estrogen receptor 1 induces coronary artery relaxation via Epac/Rap1-mediated inhibition of RhoA/Rho kinase pathway in parallel with PKA

    PubMed Central

    Yu, Xuan; Zhang, Qiao; Zhao, Yan; Schwarz, Benjamin J.; Stallone, John N.; Heaps, Cristine L.; Han, Guichun

    2017-01-01

    Previously, we reported that cAMP/PKA signaling is involved in GPER-mediated coronary relaxation by activating MLCP via inhibition of RhoA pathway. In the current study, we tested the hypothesis that activation of GPER induces coronary artery relaxation via inhibition of RhoA/Rho kinase pathway by cAMP downstream targets, exchange proteins directly activated by cAMP (Epac) as well as PKA. Our results show that Epac inhibitors, brefeldin A (BFA, 50 μM), or ESI-09 (20 μM), or CE3F4 (100 μM), all partially inhibited porcine coronary artery relaxation response to the selective GPER agonist, G-1 (0.3–3 μM); while concurrent administration of BFA and PKI (5 μM), a PKA inhibitor, almost completely blocked the relaxation effect of G-1. The Epac specific agonist, 8-CPT-2Me-cAMP (007, 1–100 μM), induced a concentration-dependent relaxation response. Furthermore, the activity of Ras-related protein 1 (Rap1) was up regulated by G-1 (1 μM) treatment of porcine coronary artery smooth muscle cells (CASMCs). Phosphorylation of vasodilator-stimulated phosphoprotein (p-VASP) was elevated by G-1 (1 μM) treatment, but not by 007 (50 μM); and the effect of G-1 on p-VASP was blocked by PKI, but not by ESI-09, an Epac antagonist. RhoA activity was similarly down regulated by G-1 and 007, whereas ESI-09 restored most of the reduced RhoA activity by G-1 treatment. Furthermore, G-1 decreased PGF2α-induced p-MYPT1, which was partially reversed with either ESI-09 or PKI; whereas, concurrent administration of ESI-09 and PKI totally prevented the inhibitory effect of G-1. The inhibitory effects of G-1 on p- MLC levels in CASMCs were mostly restored by either ESI-09 or PKI. These results demonstrate that activation of GPER induces coronary artery relaxation via concurrent inhibition of RhoA/Rho kinase by Epac/Rap1 and PKA. GPER could be a potential drug target for preventing and treating cardiovascular diseases. PMID:28278256

  15. Arabidopsis RAP2.2 plays an important role in plant resistance to Botrytis cinerea and ethylene responses.

    PubMed

    Zhao, Yang; Wei, Tong; Yin, Kang-Quan; Chen, Zhangliang; Gu, Hongya; Qu, Li-Jia; Qin, Genji

    2012-07-01

    • Ethylene plays a crucial role in plant resistance to necrotrophic pathogens, in which ETHYLENE RESPONSE FACTORs (ERFs) are often involved. • Here, we evaluated the role of an ERF transcription factor, RELATED TO AP2 2 (RAP2.2), in Botrytis resistance and ethylene responses in Arabidopsis. We analyzed the resistance of transgenic plants overexpressing RAP2.2 and the T-DNA insertion mutant to Botrytis cinerea. We assessed its role in the ethylene signaling pathway by molecular and genetic approaches. • RAP2.2-overexpressing transgenic plants showed increased resistance to B. cinerea, whereas its T-DNA insertion mutant rap2.2-3 showed decreased resistance. Overexpression of RAP2.2 in ethylene insensitive 2 (ein2) and ein3 ein3-like 1 (eil1) mutants restored their resistance to B. cinerea. Both ethylene and Botrytis infection induced the expression of RAP2.2 and the induction was disrupted in ein2 and ein3 eil1 mutants. We identified rap2.12-1 as a T-DNA insertion mutant of RAP2.12, the closest homolog of RAP2.2. The hypocotyls of rap2.2-3 rap2.12-1 double mutants showed ethylene insensitivity. The constitutive triple response in constitutive triple response1 (ctr1) was partially released in the rap2.2-3 rap2.12-1 ctr1 triple mutants. • Our findings demonstrate that RAP2.2 functions as an important regulator in Botrytis resistance and ethylene responses.

  16. 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…

  17. 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)

  18. Rap1 Stabilizes β-catenin and Enhances β-catenin-dependent Transcription and Invasion in Squamous Cell Carcinoma of the Head and Neck

    PubMed Central

    Goto, Mitsuo; Mitra, Raj S.; Liu, Min; Lee, Julia; Henson, Bradley S.; Carey, Thomas; Bradford, Carol; Prince, Mark; Wang, Cun-Yu; Fearon, Eric R.; D’Silva, Nisha J.

    2009-01-01

    Purpose In head and neck squamous cell carcinoma (HNSCC) cells, Rap1 shuttles between the nucleus and cytoplasm. Prior findings suggested that Rap1 may modulate the β-catenin-independent Wnt pathway in some settings, but the role of Rap1 in β-catenin-dependent Wnt signaling remains undefined. Experimental Design and Results We observed that β-catenin bound to active Rap1 in vitro and Rap1 activated β-catenin-TCF (T cell factor)-dependent transcription. Immunofluorescence studies showed that ectopic expression of Rap1 increased nuclear translocation of β-catenin. Overexpression of active Rap1 facilitated an increase in β-catenin-mediated transcription that was abrogated by dominant negative TCF4. Conversely, siRNA-mediated inhibition of endogenous Rap1 expression inhibited β-catenin/TCF-mediated transcription as well as invasion of HNSCC. Furthermore, inhibition of Rap1 expression downregulated the expresesion of MMP7, a transcriptional target of β-catenin/TCF. In HNSCC cells stably transfected with β-catenin or treated with lithium chloride or Wnt3A to stabilize endogenous β-catenin, inhibition of Rap1 expression led to decreases in the free pool of β-catenin. Immunohistochemical studies of tissue from HNSCC patients revealed that increased β-catenin intensity correlated with higher tumor stage. Furthermore, the prognostic effect of active Rap1 on tumor N-stage was found to depend on cytosolic β-catenin expression (p<0.013). When β-catenin is high, higher rap1GTP intensity is associated with more advanced N stage. Conclusions The findings suggest that Rap1 enhances β-catenin stability and nuclear localization. In addition to indicating that Rap1 has a significant role in regulating β-catenin and β-catenin-dependent progression to more advanced N-stage lesions, these data highlight Rap1 as a potential therapeutic target in HNSCC. PMID:20028760

  19. Histone deacetylase inhibitors upregulate Rap1GAP and inhibit Rap activity in thyroid tumor cells.

    PubMed

    Dong, Xiaoyun; Korch, Christopher; Meinkoth, Judy L

    2011-06-01

    Increases in Rap activity have been associated with tumor progression. Although activating mutations in Rap have not been described, downregulation of Rap1GAP is frequent in human tumors including thyroid carcinomas. In this study, we explored whether endogenous Rap1GAP expression could be restored to thyroid tumor cells. The effects of deacetylase inhibitors and a demethylating agent, individually and in combination, were examined in four differentiated and six anaplastic thyroid carcinoma (ATC) cell lines. Treatment with the structurally distinct histone deacetylase (HDAC) inhibitors, sodium butyrate and trichostatin A, increased Rap1GAP expression in all the differentiated thyroid carcinoma cell lines and in four of the six ATC cell lines. The demethylating agent, 5-aza-deoxycytidine, restored Rap1GAP expression in one anaplastic cell line and enhanced the effects of HDAC inhibitors in a second anaplastic cell line. Western blotting indicated that Rap2 was highly expressed in human thyroid cancer cells. Importantly, treatment with HDAC inhibitors impaired Rap2 activity in both differentiated and anaplastic tumor cell lines. The mechanism through which Rap activity is repressed appears to entail effects on the expression of multiple Rap regulators, including RapGEFs and RapGAPs. These results suggest that HDAC inhibitors may provide a tractable approach to impair Rap activity in human tumor cells.

  20. 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)

  1. 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…

  2. 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.

  3. 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

  4. 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…

  5. 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

  6. Rap Lyrics: Instruments for Language Arts Instruction.

    ERIC Educational Resources Information Center

    Jeremiah, Milford A.

    1992-01-01

    Offers suggestions for using rap lyrics to teach language skills. A brief overview of the sociocultural dimension of rap music and its literature, a theoretical orientation for the instructional methodology, and a curriculum model are presented. The prevalence of these musical forms can make them a valuable aid. (SLD)

  7. Ras-independent activation of ERK signaling via the torso receptor tyrosine kinase is mediated by Rap1.

    PubMed

    Mishra, Snigdha; Smolik, Sarah M; Forte, Michael A; Stork, Philip J S

    2005-02-22

    In Drosophila embryos, the Torso receptor tyrosine kinase (RTK) activates the small G protein Ras (D-Ras1) and the protein kinase Raf (D-Raf) to activate ERK to direct differentiation of terminal structures . However, genetic studies have demonstrated that Torso, and by extension other RTKs, can activate Raf and ERK independently of Ras . In mammalian cells, the small G protein Rap1 has been proposed to couple RTKs to ERKs. However, the ability of Rap1 to activate ERKs remains controversial, in part because direct genetic evidence supporting this hypothesis is lacking. Here, we present biochemical and genetic evidence that D-Rap1, the Drosophila homolog of Rap1, can activate D-Raf and ERK. We show that D-Rap1 binds D-Raf and activates ERKs in a GTP- and D-Raf-dependent manner. Targeted disruption of D-Rap1 expression decreased both Torso-dependent ERK activation and the ERK-dependent expression of the zygotic genes tailless and huckebein to levels similar to those achieved in D-Ras1 null embryos. Furthermore, combined deficiencies of D-Ras1 and D-Rap1 completely abolished expression of these genes, mimicking the phenotype observed in embryos lacking D-Raf. These studies provide the first direct genetic evidence of Rap1-mediated activation of the MAP kinase cascade in eukaryotic organisms.

  8. 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.

  9. 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

  10. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false What is a RAP? 270.80 Section 270.80... ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  11. 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...

  12. 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...

  13. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false What is a RAP? 270.80 Section 270.80... ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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... ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  20. 40 CFR 270.80 - What is a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false What is a RAP? 270.80 Section 270.80... ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  1. 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...

  2. 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...

  3. 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... ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.80 What is a RAP? (a) A RAP is a special form of RCRA permit that you, as an owner...

  4. 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...

  5. Infant information processing and family history of specific language impairment: converging evidence for RAP deficits from two paradigms.

    PubMed

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

    2007-03-01

    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 study is to determine if performance on infant information processing measures designed to tap RAP and global processing skills differ as a function of family history of specific language impairment (SLI) and/or the particular demand characteristics of the paradigm used. Seventeen 6- to 9-month-old infants from families with a history of specific language impairment (FH+) and 29 control infants (FH-) participated in this study. Infants' performance on two different RAP paradigms (head-turn procedure [HT] and auditory-visual habituation/recognition memory [AVH/RM]) and on a global processing task (visual habituation/recognition memory [VH/RM]) was assessed at 6 and 9 months. Toddler language and cognitive skills were evaluated at 12 and 16 months. A number of significant group differences were seen: FH+ infants showed significantly poorer discrimination of fast rate stimuli on both RAP tasks, took longer to habituate on both habituation/recognition memory measures, and had lower novelty preference scores on the visual habituation/recognition memory task. Infants' performance on the two RAP measures provided independent but converging contributions to outcome. Thus, different mechanisms appear to underlie performance on operantly conditioned tasks as compared to habituation/recognition memory paradigms. Further, infant RAP processing abilities predicted to 12- and 16-month language scores above and beyond family history of SLI. The results of this study provide additional support for the validity of infant RAP abilities as a behavioral marker for later language outcome. Finally, this is the first study to use a battery of

  6. 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.

  7. CDK5RAP3 acts as a tumor suppressor in gastric cancer through inhibition of β-catenin signaling.

    PubMed

    Wang, Jia-Bin; Wang, Zu-Wei; Li, Yun; Huang, Chao-Qun; Zheng, Chao-Hui; Li, Ping; Xie, Jian-Wei; Lin, Jian-Xian; Lu, Jun; Chen, Qi-Yue; Cao, Long-Long; Lin, Mi; Tu, Ru-Hong; Lin, Yao; Huang, Chang-Ming

    2017-01-28

    CDK5RAP3 was isolated as a binding protein of the Cdk5 activator p35. Although CDK5RAP3 has been implicated in cancer progression, its expression and function have not been investigated in gastric cancer. Our study demonstrated that the mRNA and protein levels of CDK5RAP3 were markedly decreased in gastric tumor tissues when compared with respective adjacent non-tumor tissues. CDK5RAP3 in gastric cancer cells significantly reduced cell proliferation, migration, invasion and tumor xenograft growth through inhibition of β-catenin. Secondly, CDK5RAP3 was found to suppress the phosphorylation of GSK-3β (Ser9), leading to the phosphorylation (Ser37/Thr41) and subsequent degradation of β-catenin. Lastly, the prognostic value of CDK5RAP3 for overall survival was found to be dependent on β-catenin cytoplasm/nucleus localization in human gastric cancer samples. Collectively, our results demonstrated that CDK5RAP3 negatively regulates the β-catenin signaling pathway by repressing GSK-3β phosphorylation and could be a potential therapeutic target for gastric cancer.

  8. Krit 1 interactions with microtubules and membranes are regulated by Rap1 and integrin cytoplasmic domain associated protein-1.

    PubMed

    Béraud-Dufour, Sophie; Gautier, Romain; Albiges-Rizo, Corinne; Chardin, Pierre; Faurobert, Eva

    2007-11-01

    The small G protein Rap1 regulates diverse cellular processes such as integrin activation, cell adhesion, cell-cell junction formation and cell polarity. It is crucial to identify Rap1 effectors to better understand the signalling pathways controlling these processes. Krev interaction trapped 1 (Krit1), a protein with FERM (band four-point-one/ezrin/radixin/moesin) domain, was identified as a Rap1 partner in a yeast two-hybrid screen, but this interaction was not confirmed in subsequent studies. As the evidence suggests a role for Krit1 in Rap1-dependent pathways, we readdressed this question. In the present study, we demonstrate by biochemical assays that Krit1 interacts with Rap1A, preferentially its GTP-bound form. We show that, like other FERM proteins, Krit1 adopts two conformations: a closed conformation in which its N-terminal NPAY motif interacts with its C-terminus and an opened conformation bound to integrin cytoplasmic domain associated protein (ICAP)-1, a negative regulator of focal adhesion assembly. We show that a ternary complex can form in vitro between Krit1, Rap1 and ICAP-1 and that Rap1 binds the Krit1 FERM domain in both closed and opened conformations. Unlike ICAP-1, Rap1 does not open Krit1. Using sedimentation assays, we show that Krit1 binds in vitro to microtubules through its N- and C-termini and that Rap1 and ICAP-1 inhibit Krit1 binding to microtubules. Consistently, YFP-Krit1 localizes on cyan fluorescent protein-labelled microtubules in baby hamster kidney cells and is delocalized from microtubules upon coexpression with activated Rap1V12. Finally, we show that Krit1 binds to phosphatidylinositol 4,5-P(2)-containing liposomes and that Rap1 enhances this binding. Based on these results, we propose a model in which Krit1 would be delivered by microtubules to the plasma membrane where it would be captured by Rap1 and ICAP-1.

  9. 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...

  10. 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.

  11. 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.

  12. The rap GTPases regulate B cell morphology, immune-synapse formation, and signaling by particulate B cell receptor ligands.

    PubMed

    Lin, Kevin B L; Freeman, Spencer A; Zabetian, Saba; Brugger, Hayley; Weber, Michele; Lei, Victor; Dang-Lawson, May; Tse, Kathy W K; Santamaria, Rene; Batista, Facundo D; Gold, Michael R

    2008-01-01

    B lymphocytes spread and extend membrane processes when searching for antigens and form immune synapses upon contacting cells that display antigens on their surface. Although these dynamic morphological changes facilitate B cell activation, the signaling pathways underlying these processes are not fully understood. We found that activation of the Rap GTPases was essential for these changes in B cell morphology. Rap activation was important for B cell receptor (BCR)- and lymphocyte-function-associated antigen-1 (LFA-1)-induced spreading, for BCR-induced immune-synapse formation, and for particulate BCR ligands to induce localized F-actin assembly and membrane-process extension. Rap activation and F-actin assembly were also required for optimal BCR signaling in response to particulate antigens but not soluble antigens. Thus by controlling B cell morphology and cytoskeletal organization, Rap might play a key role in the activation of B cells by particulate and cell-associated antigens.

  13. 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

  14. 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…

  15. 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.

  16. 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.

  17. 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.…

  18. E-cadherin endocytosis regulates the activity of Rap1: a traffic light GTPase at the crossroads between cadherin and integrin function.

    PubMed

    Balzac, Fiorella; Avolio, Maria; Degani, Simona; Kaverina, Irina; Torti, Mauro; Silengo, Lorenzo; Small, J Victor; Retta, Saverio Francesco

    2005-10-15

    The coordinate modulation of cadherin and integrin functions plays an essential role in fundamental physiological and pathological processes, including morphogenesis and cancer. However, the molecular mechanisms underlying the functional crosstalk between cadherins and integrins are still elusive. Here, we demonstrate that the small GTPase Rap1, a crucial regulator of the inside-out activation of integrins, is a target for E-cadherin-mediated outside-in signaling. In particular, we show that a strong activation of Rap1 occurs upon adherens junction disassembly that is triggered by E-cadherin internalization and trafficking along the endocytic pathway. By contrast, Rap1 activity is not influenced by integrin outside-in signaling. Furthermore, we demonstrate that the E-cadherin endocytosis-dependent activation of Rap1 is associated with and controlled by an increased Src kinase activity, and is paralleled by the colocalization of Rap1 and E-cadherin at the perinuclear Rab11-positive recycling endosome compartment, and the association of Rap1 with a subset of E-cadherin-catenin complexes that does not contain p120ctn. Conversely, Rap1 activity is suppressed by the formation of E-cadherin-dependent cell-cell junctions as well as by agents that inhibit either Src activity or E-cadherin internalization and intracellular trafficking. Finally, we demonstrate that the E-cadherin endocytosis-dependent activation of Rap1 is associated with and is required for the formation of integrin-based focal adhesions. Our findings provide the first evidence of an E-cadherin-modulated endosomal signaling pathway involving Rap1, and suggest that cadherins may have a novel modulatory role in integrin adhesive functions by fine-tuning Rap1 activation.

  19. Rap1 overexpression reveals that activated RasD induces separable defects during Dictyostelium development.

    PubMed

    Louis, S A; Weeks, G; Spiegelman, G B

    1997-10-15

    One of the Dictyostelium ras genes, rasD, is expressed preferentially in prestalk cells at the slug stage of development and overexpression of this gene containing a G12T activating mutation causes the formation of aberrant multitipped aggregates that are blocked from further development (Reymond et al., 1986, Nature, 323, 340-343). The ability of the Dictyostelium rap1 gene to suppress this abnormal developmental phenotype was investigated. The rap1 gene and G12V activated and G10V negative mutant forms of the rap1 gene were independently linked to the rasD promoter and each construct used to transform M1, a Dictyostelium cell line expressing RasD[G12T]. Transformants of M1 that expressed Rap1 or Rap1[G12V] protein still formed multitipped aggregates, but most tips were able to complete development and form fruiting bodies. Cell lines showing this modified phenotype were designated ME (multitipped escape). The rap1[G10V] construct did not modify the M1 phenotype. These data suggest that overexpression of RasD[G12T] has two effects, the formation of a multitipped aggregate and a block in subsequent differentiation and that the expression of Rap1 or Rap1[G12V] reverses only the latter. Differentiation of ME cells in low density monolayers showed the identical low level of stalk and spore cell formation seen for M1 cells under the same conditions. Thus the cell autonomous defect in monolayer differentiation induced in the M1 strain was not corrected in the ME strain. Cell type-specific gene expression during the development of M1 cells is dramatically altered: prestalk cell-specific gene expression is greatly enhanced, whereas prespore-specific gene expression is almost suppressed (Louis et al., 1997, Mol. Biol. Cell, 8, 303-312). During the development of ME cells, ecmA mRNA levels were restored to those seen for Ax3, and tagB mRNA levels were also markedly reduced, although not to Ax3 levels. cotC expression in ME cells was enhanced severalfold relative to M1

  20. Strigolactones' ability to regulate root development may be executed by induction of the ethylene pathway.

    PubMed

    Koltai, Hinanit

    2011-07-01

    The newly defined phytohormones strigolactones (SLs) were recently shown to act as regulators of root development. Their positive effect on root-hair (RH) elongation enabled examination of their cross talk with auxin and ethylene. Analysis of wild-type plants and hormone-signaling mutants combined with hormonal treatments suggested that SLs and ethylene regulate RH elongation via a common regulatory pathway, in which ethylene is epistatic to SLs. The SL and auxin hormonal pathways were suggested to converge for regulation of RH elongation; this convergence was suggested to be mediated via the ethylene pathway, and to include regulation of auxin transport.

  1. 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.

  2. 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...

  3. 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…

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. 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

  9. 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…

  10. An Atypical Phr Peptide Regulates the Developmental Switch Protein RapH ▿ †

    PubMed Central

    Mirouze, Nicolas; Parashar, Vijay; Baker, Melinda D.; Dubnau, David A.; Neiditch, Matthew B.

    2011-01-01

    Under conditions of nutrient limitation and high population density, the bacterium Bacillus subtilis can initiate a variety of developmental pathways. The signaling systems regulating B. subtilis differentiation are tightly controlled by switch proteins called Raps, named after the founding members of the family, which were shown to be response regulator aspartate phosphatases. A phr gene encoding a secreted pentapeptide that regulates the activity of its associated Rap protein was previously identified downstream of 8 of the chromosomally encoded rap genes. We identify and validate here the sequence of an atypical Phr peptide, PhrH, by in vivo and in vitro analyses. Using a luciferase reporter bioassay combined with in vitro experiments, we found that PhrH is a hexapeptide (TDRNTT), in contrast to the other characterized Phr pentapeptides. We also determined that phrH expression is driven by a promoter lying within rapH. Unlike the previously identified dedicated σH-driven phr promoters, it appears that phrH expression most likely requires σA. Furthermore, we show that PhrH can antagonize both of the known activities of RapH: the dephosphorylation of Spo0F and the sequestration of ComA, thus promoting the development of spores and the competent state. Finally, we propose that PhrH is the prototype of a newly identified class of Phr signaling molecules consisting of six amino acids. This class likely includes PhrI, which regulates RapI and the expression, excision, and transfer of the mobile genetic element ICEBs1. PMID:21908671

  11. Dissection of RAP-LRP interactions: binding of RAP and RAP fragments to complement-like repeats 7 and 8 from ligand binding cluster II of LRP.

    PubMed

    Lazic, Ana; Dolmer, Klavs; Strickland, Dudley K; Gettins, Peter G W

    2006-06-15

    The receptor associated protein (RAP) is a three domain 38kDa ER-resident chaperone that helps folding of LRP and other LDL receptor family members and prevents premature binding of protein ligands. It competes strongly with all known LRP ligands. To further understanding of the specificity of RAP-LRP interactions, the binding of RAP and RAP fragments to two domains (CR7-CR8) from one of the main ligand-binding regions of LRP has been examined by 2D HSQC NMR spectroscopy and isothermal titration calorimetry. We found that RAP contains two binding sites for CR7-CR8, with the higher affinity site (K(d) approximately 1microM) located in the C-terminal two-thirds and the weaker site (K(d) approximately 5microM) in the N-terminal third of RAP. Residues from both CR7 and CR8 are involved in binding at each RAP site. The presence of more than one binding site on RAP for CR domains from LRP, together with the previous demonstration by others that RAP can bind to CR5-CR6 with comparably low affinities suggest an explanation for the dual roles of RAP as a folding chaperone and a tight competitive inhibitor of ligand binding.

  12. Pathways to fraction learning: Numerical abilities mediate the relation between early cognitive competencies and later fraction knowledge.

    PubMed

    Ye, Ai; Resnick, Ilyse; Hansen, Nicole; Rodrigues, Jessica; Rinne, Luke; Jordan, Nancy C

    2016-12-01

    The current study investigated the mediating role of number-related skills in the developmental relationship between early cognitive competencies and later fraction knowledge using structural equation modeling. Fifth-grade numerical skills (i.e., whole number line estimation, non-symbolic proportional reasoning, multiplication, and long division skills) mapped onto two distinct factors: magnitude reasoning and calculation. Controlling for participants' (N=536) demographic characteristics, these two factors fully mediated relationships between third-grade general cognitive competencies (attentive behavior, verbal and nonverbal intellectual abilities, and working memory) and sixth-grade fraction knowledge (concepts and procedures combined). However, specific developmental pathways differed by type of fraction knowledge. Magnitude reasoning ability fully mediated paths from all four cognitive competencies to knowledge of fraction concepts, whereas calculation ability fully mediated paths from attentive behavior and verbal ability to knowledge of fraction procedures (all with medium to large effect sizes). These findings suggest that there are partly overlapping, yet distinct, developmental pathways from cognitive competencies to general fraction knowledge, fraction concepts, and fraction procedures.

  13. Efficient localization of synchronous EEG source activities using a modified RAP-MUSIC algorithm.

    PubMed

    Liu, Hesheng; Schimpf, Paul H

    2006-04-01

    Synchronization across different brain regions is suggested to be a possible mechanism for functional integration. Noninvasive analysis of the synchronization among cortical areas is possible if the electrical sources can be estimated by solving the electroencephalography inverse problem. Among various inverse algorithms, spatio-temporal dipole fitting methods such as RAP-MUSIC and R-MUSIC have demonstrated superior ability in the localization of a restricted number of independent sources, and also have the ability to reliably reproduce temporal waveforms. However, these algorithms experience difficulty in reconstructing multiple correlated sources. Accurate reconstruction of correlated brain activities is critical in synchronization analysis. In this study, we modified the well-known inverse algorithm RAP-MUSIC to a multistage process which analyzes the correlation of candidate sources and searches for independent topographies (ITs) among precorrelated groups. Comparative studies were carried out on both simulated data and clinical seizure data. The results demonstrated superior performance with the modified algorithm compared to the original RAP-MUSIC in recovering synchronous sources and localizing the epileptiform activity. The modified RAP-MUSIC algorithm, thus, has potential in neurological applications involving significant synchronous brain activities.

  14. 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…

  15. 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…

  16. 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.

  17. Structural basis of response regulator dephosphorylation by Rap phosphatases.

    PubMed

    Parashar, Vijay; Mirouze, Nicolas; Dubnau, David A; Neiditch, Matthew B

    2011-02-08

    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 β4-α4 loop adopts an active-site proximal conformation.

  18. 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.

  19. The small GTPases Ras and Rap1 bind to and control TORC2 activity

    PubMed Central

    Khanna, Ankita; Lotfi, Pouya; Chavan, Anita J.; Montaño, Nieves M.; Bolourani, Parvin; Weeks, Gerald; Shen, Zhouxin; Briggs, Steven P.; Pots, Henderikus; Van Haastert, Peter J. M.; Kortholt, Arjan; Charest, Pascale G.

    2016-01-01

    Target of Rapamycin Complex 2 (TORC2) has conserved roles in regulating cytoskeleton dynamics and cell migration and has been linked to cancer metastasis. However, little is known about the mechanisms regulating TORC2 activity and function in any system. In Dictyostelium, TORC2 functions at the front of migrating cells downstream of the Ras protein RasC, controlling F-actin dynamics and cAMP production. Here, we report the identification of the small GTPase Rap1 as a conserved binding partner of the TORC2 component RIP3/SIN1, and that Rap1 positively regulates the RasC-mediated activation of TORC2 in Dictyostelium. Moreover, we show that active RasC binds to the catalytic domain of TOR, suggesting a mechanism of TORC2 activation that is similar to Rheb activation of TOR complex 1. Dual Ras/Rap1 regulation of TORC2 may allow for integration of Ras and Rap1 signaling pathways in directed cell migration. PMID:27172998

  20. 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

  1. 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

    Figueroa-Yañez, Luis; Pereira-Santana, Alejandro; 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.

  2. 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

  3. 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...

  4. 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...

  5. 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 §...

  6. 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....

  7. 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...

  8. 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 §...

  9. 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...

  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 270.210 - 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...

  12. 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 §...

  13. 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...

  14. 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...

  15. 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....

  16. 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...

  17. 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...

  18. 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,...

  19. 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....

  20. 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...

  1. 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...

  2. 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 §...

  3. 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....

  4. 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 §...

  5. 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...

  6. 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,...

  7. 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...

  8. 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...

  9. 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....

  10. Targeted disruption of N-RAP gene function by RNA interference: a role for N-RAP in myofibril organization.

    PubMed

    Dhume, Ashwini; Lu, Shajia; Horowits, Robert

    2006-08-01

    N-RAP is a muscle-specific protein concentrated in myofibril precursors during sarcomere assembly and at intercalated disks in adult heart. We used RNA interference to achieve a targeted decrease in N-RAP transcript and protein levels in primary cultures of embryonic mouse cardiomyocytes. N-RAP transcript levels were decreased by approximately 70% within 2 days following transfection with N-RAP specific siRNA. N-RAP protein levels steadily decreased over several days, reaching approximately 50% of control levels within 6 days. N-RAP protein knockdown was associated with decreased myofibril assembly, as assessed by alpha-actinin organization into mature striations. Transcripts encoding N-RAP binding proteins associated with assembling or mature myofibrils, such as alpha-actinin, Krp1, and muscle LIM protein, were expressed at normal levels during N-RAP protein knockdown, and alpha-actinin and Krp-1 protein levels were also unchanged. Transcripts encoding muscle myosin heavy chain and nonmuscle myosin heavy chain IIB were also expressed at relatively normal levels. However, decreased N-RAP protein levels were associated with dramatic changes in the encoded myosin proteins, with muscle myosin heavy chain levels increasing and nonmuscle myosin heavy chain IIB decreasing. N-RAP transcript and protein levels recovered to normal by days 6 and 7, respectively, and the changes in myofibril organization and myosin heavy chain isoform levels were reversed. Our data indicate that we can achieve transient N-RAP protein knockdown using the RNA interference technique and that alpha-actinin organization into myofibrils in cardiomyocytes is closely linked to N-RAP protein levels. Finally, N-RAP protein levels regulate the balance between nonmuscle myosin IIB and muscle myosin by post-trancriptional mechanisms.

  11. 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

  12. Tetrahydroxystilbene glucoside improves learning and (or) memory ability of aged rats and may be connected to the APP pathway.

    PubMed

    Hou, Ying; Yang, Qidong; Zhou, Lin; Du, Xiaoping; Li, Min; Yuan, Mei; Zhou, Zhiwen; Li, Zhenguo

    2011-11-01

    The aim of this study is to evaluate the protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) on learning and (or) memory deficit in aged rats, as well as to explore the possible connection between TSG and the β-amyloid precursor protein (APP) pathway. Sprague-Dawley rats were randomly divided into a young control group (age, 4 months), an aged control group (age, 22 months), and a TSG-treated group (age, 22 months). TSG at doses of 50 mg·kg(-1)·day(-1) was intragastrically administered to 22-month-old rats for 4 weeks. The learning and (or) memory ability was measured using the Morris water maze (MWM) test, and the mRNA and protein expression of APP pathway proteins was measured by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. The aged rats exhibited obvious learning and (or) memory deficit when compared with the young rats, but TSG treatment significantly improved the learning and (or) memory ability in the aged rats, as noted from the MWM test. RT-PCR and Western blot analysis showed an increase in the expression of beta-site APP cleaving enzyme 1 (BACE1) and A Disintegrin And Metalloproteinase 17 (ADAM17) in aged rats, and a decrease in ADAM10; however, TSG treatment significantly increased the mRNA and protein expression of ADAM10 (p < 0.01, compared with aged control rats). These results provide solid evidence for the therapeutic effect of TSG on age-related cognitive impairment, especially spatial learning and memory deficit. TSG might exert this effect through the APP pathway, although further studies on the topic are required.

  13. Possible roles of the cAMP-mediators EPAC and RAP1 in decidualization of rat uterus.

    PubMed

    Kusama, Kazuya; Yoshie, Mikihiro; Tamura, Kazuhiro; Daikoku, Takiko; Takarada, Tsutomu; Tachikawa, Eiichi

    2014-06-01

    The optimal decidualization of endometrial stromal cells (ESCs) following embryo implantation is one of the critical steps to establish pregnancy in rodents and humans. This step is intricately regulated by ovarian hormones. Using in vitro human ESCs model, we previously showed that activation of a cAMP mediator, exchange protein directly activated by cAMP (EPAC), promotes ovarian steroid- or cAMP analog-induced decidualization. However, expressions and functions of EPAC and RAP1 in the uterus during pregnancy have not yet been examined. In this study, we found that the expression of EPAC2 and RAP1 was markedly upregulated in the decidual cells at the implantation sites on days 7 and 9 of pregnancy in rats. Furthermore, both delayed-implantation and artificial decidualization models showed that EPAC2 and RAP1 expression was enhanced in decidual cells. Significant activation of cAMP-responsive element-binding protein (CREB), a central transcriptional factor of cAMP signaling, was observed in decidual cells. These spatiotemporal expressions of protein related EPAC pathway are overlapped by sites with activated cAMP signaling, indicating the association of EPAC signaling with decidualization. Strikingly, further studies in in vitro rat decidualization model showed that the cAMP analog and medroxyprogesterone stimulated the expression of decidual markers, while knockdown of EPAC1/2 and RAP1 attenuated the expressions of these markers. Together, these findings suggest that EPAC and RAP1 are the crucial factors for endometrial decidualization in rat pregnancy.

  14. Identification and biochemical characterization of Rap2C, a new member of the Rap family of small GTP-binding proteins.

    PubMed

    Paganini, Simona; Guidetti, Gianni Francesco; Catricalà, Silvia; Trionfini, Piera; Panelli, Simona; Balduini, Cesare; Torti, Mauro

    2006-01-01

    The Rap family of small GTP-binding proteins is composed by four different members: Rap1A, Rap1B, Rap2A and Rap2B. In this work we report the identification and characterization of a fifth member of this family of small GTPases. This new protein is highly homologous to Rap2A and Rap2B, binds labeled GTP on nitrocellulose, and is recognized by a specific anti-Rap2 antibody, but not by an anti-Rap1 antibody. The protein has thus been named Rap2C. Binding of GTP to recombinant purified Rap2C was Mg(2+)-dependent. However, accurate comparison of the kinetics of nucleotide binding and release revealed that Rap2C bound GTP less efficiently and possessed slower rate of GDP release compared to the highly homologous Rap2B. Moreover, in the presence of Mg(2+), the relative affinity of Rap2C for GTP was only about twofold higher than that for GDP, while, under the same conditions, Rap2B was able to bind GTP with about sevenfold higher affinity than GDP. When expressed in eukaryotic cells, Rap2C localized at the plasma membrane, as dictated by the presence of a CAAX motif at the C-terminus. We found that Rap2C represented the predominant Rap2 protein expressed in circulating mononuclear leukocytes, but was not present in platelets. Importantly, Rap2C was found to be expressed in human megakaryocytes, suggesting that the protein may be down-regulated during platelets generation. This work demonstrates that Rap2C is a new member of the Rap2 subfamily of proteins, able to bind guanine nucleotides with peculiar properties, and differently expressed by various hematopoietic subsets. This new protein may therefore contribute to the still poorly clarified cellular events regulated by this subfamily of GTP-binding proteins.

  15. 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.

  16. 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

  17. 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…

  18. 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

    2016-02-03

    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.

  19. 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…

  20. 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.…

  1. A Systematic Review on the Functions of Rap Among Gangs.

    PubMed

    Lozon, Jeffrey; Bensimon, Moshe

    2015-11-27

    Although the field of gangs is well studied, information regarding the way gangs may use or misuse music for different needs is sparse. The aim of this systematic review is to gather descriptive and empirical information to ascertain the important roles rap music possesses within gang life. This review suggests five main functions of rap used within gangs with an emphasis on the subgenre of gangsta rap. First, rap facilitates antisocial behavior by reinforcing such messages in its lyrics. Second, its deviant lyrics serve as a reflection of the violent reality experienced in many urban ghetto communities. Third, it operates as a means for constructing individual and collective identity, as well as resistance identity. Fourth, it functions as an educating force by teaching its members how to act and respond in the urban ghetto. Finally, rap glorifies gang norms among newcomers and successfully spreads its values to the general population.

  2. 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

  3. 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…

  4. 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

  5. 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.

  6. RAP30/74: a general initiation factor that binds to RNA polymerase II.

    PubMed Central

    Burton, Z F; Killeen, M; Sopta, M; Ortolan, L G; Greenblatt, J

    1988-01-01

    We have previously shown by affinity chromatography that RAP30 and RAP74 are the mammalian proteins that have the highest affinity for RNA polymerase II. Here we show that RAP30 binds to RAP74 and that the RAP30-RAP74 complex (RAP30/74) is required for accurate initiation by RNA polymerase II. RAP30/74 is required for accurate transcription from the following promoters: the adenovirus major late promoter, the long terminal repeat of human immunodeficiency virus, P2 of the human c-myc gene, the mouse beta maj-globin promoter (all of which have TATA boxes), and the mouse dihydrofolate reductase promoter (which lacks a TATA box). RAP30/74 is not required for initiation by RNA polymerase III at the adenovirus virus-associated RNA promoters. Therefore, RAP30/74 is a general initiation factor that binds to RNA polymerase II. Images PMID:3380090

  7. Rap1 Ameliorates Renal Tubular Injury in Diabetic Nephropathy

    PubMed Central

    Xiao, Li; Zhu, Xuejing; Yang, Shikun; Liu, Fuyou; Zhou, Zhiguang; Zhan, Ming; Xie, Ping; Zhang, Dongshan; Li, Jun; Song, Panai; Kanwar, Yashpal S.; Sun, Lin

    2014-01-01

    Rap1b ameliorates high glucose (HG)-induced mitochondrial dysfunction in tubular cells. However, its role and precise mechanism in diabetic nephropathy (DN) in vivo remain unclear. We hypothesize that Rap1 plays a protective role in tubular damage of DN by modulating primarily the mitochondria-derived oxidative stress. The role and precise mechanisms of Rap1b on mitochondrial dysfunction and of tubular cells in DN were examined in rats with streptozotocin (STZ)-induced diabetes that have Rap1b gene transfer using an ultrasound microbubble-mediated technique as well as in renal proximal epithelial tubular cell line (HK-2) exposed to HG ambiance. The results showed that Rap1b expression decreased significantly in tubules of renal biopsies from patients with DN. Overexpression of a constitutively active Rap1b G12V notably ameliorated renal tubular mitochondrial dysfunction, oxidative stress, and apoptosis in the kidneys of STZ-induced rats, which was accompanied with increased expression of transcription factor C/EBP-β and PGC-1α. Furthermore, Rap1b G12V also decreased phosphorylation of Drp-1, a key mitochondrial fission protein, while boosting the expression of genes related to mitochondrial biogenesis and antioxidants in HK-2 cells induced by HG. These effects were imitated by transfection with C/EBP-β or PGC-1α short interfering RNA. In addition, Rap1b could modulate C/EBP-β binding to the endogenous PGC-1α promoter and the interaction between PGC-1α and catalase or mitochondrial superoxide dismutase, indicating that Rap1b ameliorates tubular injury and slows the progression of DN by modulation of mitochondrial dysfunction via C/EBP-β–PGC-1α signaling. PMID:24353183

  8. Characterize RAP80, a Potential Tumor Suppressor Gene

    DTIC Science & Technology

    2008-04-01

    sites of DNA damage (21–23). One of them is Fanconi anemia complementation group D2 ( FANCD2 ). However, RAP80 foci still form normally after irradiation...in FANCD2 -deficient cells (fig. S7), suggesting that there may be other as-yet-unidentified ubiquitinated proteins that act RAP80WT RAP80D1...Horazdovsky for providing constructs encoding HSJ1A and Ubi-GST, respectively; and A. D’Andrea for providing FANCD2 -deficient and FANCD2 -reconstituted

  9. Rebellious Rhapsody: Metal, Rap, Community, and Individuation.

    PubMed

    Reddick, Brad H.; Beresin, Eugene V.

    2002-03-01

    Music can be a powerful force and tool in the life of an adolescent. It forms a social context and informs the adolescent about the adult world through the lens of artists' lives, language, and presence as models. Allegiance to a form of music is allegiance to those who make it, a way to friendship and kinship, and a road to personal identity through belonging. In their relationships formed through music, teens can create a sense of community that may be lacking in the life of family. The rebellious music of earlier generations has given rise to complex musical genres, rap and heavy metal, that are strong in defiance and controversial in their violent and sexual content. What do these musical affiliations tell us about certain segments of adolescent development and culture? The authors consider this question by exploring the form and content of the music while using it to illuminate psychodynamic and psychosocial aspects of adolescent development.

  10. "Everybody Gotta Have a Dream": Rap-centered Aspirations among Young Black Males Involved in Rap Music Production - A Qualitative Study.

    PubMed

    Foster, B Brian

    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.

  11. 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.

  12. Rap1 and integrin inside-out signaling.

    PubMed

    Katagiri, Koko; Kinashi, Tatsuo

    2012-01-01

    In leukocytes, integrins play important roles in adhesive interactions with endothelium, antigen-presenting cells, and effector functions such as cytotoxicity. This chapter describes methods to study Ras proximity 1 (Rap1), a signaling molecule that has been increasingly recognized as an important regulator of integrin-mediated cell adhesion in the immune system as well as hemostasis. Rap1 is activated by a wide variety of external stimuli including chemokines and antigens. Signaling via Rap1 transmits an inside-out signal to the integrins, thereby increasing adhesiveness to ligands such as immunoglobulin superfamily proteins as well as extracellular matrix proteins and plasma proteins. This process induces leukocyte cell adhesion to the endothelium and antigen-presenting cells. In addition to integrin regulation, activated Rap1 induces cell polarity of lymphocytes, which is coordinated with LFA-1 redistribution to the leading edge.

  13. Rap GTPase-mediated adhesion and migration: A target for limiting the dissemination of B-cell lymphomas?

    PubMed

    Lin, Kevin B L; Freeman, Spencer A; Gold, Michael R

    2010-01-01

    B-cell lymphomas, which arise in lymphoid organs, can spread rapidly via the circulatory system and form solid tumors within multiple organs. Rate-limiting steps in this metastatic process may be the adhesion of lymphoma cells to vascular endothelial cells, their exit from the vasculature and their migration to tissue sites that will support tumor growth. Thus proteins that control B cell adhesion and migration are likely to be key factors in lymphoma dissemination, and hence potential targets for therapeutic intervention. The Rap GTPases are master regulators of integrin activation, cell motility and the underlying cytoskeletal, adhesion and membrane dynamics. We have recently shown that Rap activation is critical for B-lymphoma cells to undergo transendothelial migration in vitro and in vivo. As a consequence, suppressing Rap activation impairs the ability of intravenously injected B-lymphoma cells to form solid tumors in the liver and other organs. We discuss this work in the context of targeting Rap, its downstream effectors, or other regulators of B cell adhesion and migration as an approach for limiting the dissemination of B-lymphoma cells and the development of secondary tumors.

  14. 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...

  15. 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...

  16. 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...

  17. 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

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 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...

  3. 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...

  4. PhotoRApToR: PHOTOmetric Research APplication TO Redshifts

    NASA Astrophysics Data System (ADS)

    Brescia, Massimo; Cavuoti, Stefano

    2014-08-01

    PhotoRApToR (PHOTOmetric Research APplication TO Redshifts) solves regression and classification problems and is specialized for photo-z estimation. PhotoRApToR offers data table manipulation capabilities and 2D and 3D graphics tools for data visualization; it also provides a statistical report for both classification and regression experiments. The code is written in Java; the machine learning model is in C++ to increase the core execution speed.

  5. Rap video vs. traditional video for teaching nutrition.

    PubMed

    Connelly, J O; Berryman, T; Tolley, E A

    1996-01-01

    This study compared the effectiveness of a rap video with a traditional video in providing nutrition information. Sixty pregnant African-American females (ages 14 through 18) were randomly assigned to view either a rap video or a traditional video about good nutrition. The data revealed no significant difference in scores between the two versions; both videos produced significant learning; and 17 and 18 year olds scored higher than 15 and 16 year olds.

  6. 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.

  7. Ras and Rap signaling in synaptic plasticity and mental disorders.

    PubMed

    Stornetta, Ruth L; Zhu, J Julius

    2011-02-01

    The Ras family GTPases (Ras, Rap1, and Rap2) and their downstream mitogen-activated protein kinases (ERK, JNK, and p38MAPK) and PI3K signaling cascades control various physiological processes. In neuronal cells, recent studies have shown that these parallel cascades signal distinct forms of AMPA-sensitive glutamate receptor trafficking during experience-dependent synaptic plasticity and adaptive behavior. Interestingly, both hypo- and hyperactivation of Ras/ Rap signaling impair the capacity of synaptic plasticity, underscoring the importance of a "happy-medium" dynamic regulation of the signaling. Moreover, accumulating reports have linked various genetic defects that either up- or down-regulate Ras/Rap signaling with several mental disorders associated with learning disability (e.g., Alzheimer's disease, Angelman syndrome, autism, cardio-facio-cutaneous syndrome, Coffin-Lowry syndrome, Costello syndrome, Cowden and Bannayan-Riley-Ruvalcaba syndromes, fragile X syndrome, neurofibromatosis type 1, Noonan syndrome, schizophrenia, tuberous sclerosis, and X-linked mental retardation), highlighting the necessity of happy-medium dynamic regulation of Ras/Rap signaling in learning behavior. Thus, the recent advances in understanding of neuronal Ras/Rap signaling provide a useful guide for developing novel treatments for mental diseases.

  8. A crucial role for DOK1 in PDGF-BB-stimulated glioma cell invasion through p130Cas and Rap1 signalling.

    PubMed

    Barrett, Angela; Evans, Ian M; Frolov, Antonina; Britton, Gary; Pellet-Many, Caroline; Yamaji, Maiko; Mehta, Vedanta; Bandopadhyay, Rina; Li, Ningning; Brandner, Sebastian; Zachary, Ian C; Frankel, Paul

    2014-06-15

    DOK1 regulates platelet-derived growth factor (PDGF)-BB-stimulated glioma cell motility. Mechanisms regulating tumour cell motility are essential for invasion and metastasis. We report here that PDGF-BB-mediated glioma cell invasion and migration are dependent on the adaptor protein downstream of kinase 1 (DOK1). DOK1 is expressed in several glioma cell lines and in tumour biopsies from high-grade gliomas. DOK1 becomes tyrosine phosphorylated upon PDGF-BB stimulation of human glioma cells. Knockdown of DOK1 or expression of a DOK1 mutant (DOK1FF) containing Phe in place of Tyr at residues 362 and 398, resulted in inhibition of both the PDGF-BB-induced tyrosine phosphorylation of p130Cas (also known as BCAR1) and the activation of Rap1. DOK1 colocalises with tyrosine phosphorylated p130Cas at the cell membrane of PDGF-BB-treated cells. Expression of a non-tyrosine-phosphorylatable substrate domain mutant of p130Cas (p130Cas15F) inhibited PDGF-BB-mediated Rap1 activation. Knockdown of DOK1 and Rap1 inhibited PDGF-BB-induced chemotactic cell migration, and knockdown of DOK1 and Rap1 and expression of DOK1FF inhibited PDGF-mediated three-dimensional (3D) spheroid invasion. These data show a crucial role for DOK1 in the regulation of PDGF-BB-mediated tumour cell motility through a p130Cas-Rap1 signalling pathway. [Corrected

  9. DLL4 overexpression increases gastric cancer stem/progenitor cell self-renewal ability and correlates with poor clinical outcome via Notch-1 signaling pathway activation.

    PubMed

    Miao, Zhi-Feng; Xu, Hao; Xu, Hui-Mian; Wang, Zhen-Ning; Zhao, Ting-Ting; Song, Yong-Xi; Xu, Ying-Ying

    2017-01-01

    Gastric cancer is one of the most common malignant diseases, and poses a serious threat to the quality of human life. Gastric cancer stem/progenitor cells (GCSPCs) have critical effects on tumor formation, affecting specific features of self-renewal and differentiation and playing a critical role in metastasis. The Notch-1 pathway is crucially important to GCSPCs and is regulated by DLL4. In this study, DLL4 and Nestin levels were measured in 383 gastric cancer tissue samples by immunohistochemistry, and the clinico-pathological features of patients assessed. After DLL4 silencing in selected gastric cancer cell lines, the expression of GCSPC markers and colony formation ability were analyzed and the self-renewal and differentiation capacities of the cells were evaluated. The relationship between DLL4 levels and Notch-1 signaling pathway effector amounts was assessed via Western blotting and immunofluorescence. Finally, the tumor formation ability of the gastric cancer cells was evaluated with different levels of DLL4 and multiple cell densities in vivo. Our results indicate that DLL4 expression is associated with TNM stage and cancer metastasis, with high amounts of DLL4 leading to poor outcome. DLL4 silencing inhibited the self-renewal ability of GCSPCs and increased their multidifferentiation capacity, resulting in reduced GCSPC ratios. DLL4 knockdown also blocked the Notch-1 pathway, weakening invasion ability and resistance to 5-FU chemotherapy. In vivo, DLL4 silencing inhibited the tumor formation ability of GCSPCs. In conclusion, DLL4 affects GCSPC stemness, altering their pathological behavior. DLL4 silencing inhibits GCSPC metastatic potential both in vitro and in vivo by impeding Notch-1 signaling pathway activation, indicating that DLL4 may be a new potential therapeutic target.

  10. 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.

  11. Regulation of vascular endothelial junction stability and remodeling through Rap1-Rasip1 signaling.

    PubMed

    Wilson, Christopher W; Ye, Weilan

    2014-01-01

    The ability of blood vessels to sense and respond to stimuli such as fluid flow, shear stress, and trafficking of immune cells is critical to the proper function of the vascular system. Endothelial cells constantly remodel their cell-cell junctions and the underlying cytoskeletal network in response to these exogenous signals. This remodeling, which depends on regulation of the linkage between actin and integral junction proteins, is controlled by a complex signaling network consisting of small G proteins and their various downstream effectors. In this commentary, we summarize recent developments in understanding the small G protein RAP1 and its effector RASIP1 as critical mediators of endothelial junction stabilization, and the relationship between RAP1 effectors and modulation of different subsets of endothelial junctions.   The vasculature is a dynamic organ that is constantly exposed to a variety of signaling stimuli and mechanical stresses. In embryogenesis, nascent blood vessels form via a process termed vasculogenesis, wherein mesodermally derived endothelial precursor cells aggregate into cords, which subsequently form a lumen that permits trafficking of plasma and erythrocytes. (1)(,) (2) Angiogenesis occurs after establishment of this primitive vascular network, where new vessels sprout from existing vessels, migrate into newly expanded tissues, and anastomose to form a functional and complex circulatory network. (1)(,) (2) In the mouse, this process occurs through the second half of embryogenesis and into postnatal development in some tissues, such as the developing retinal vasculature. (3) Further, angiogenesis occurs in a variety of pathological conditions, such as diabetic retinopathy, age-related macular degeneration, inflammatory diseases such as rheumatoid arthritis, wound healing, and tumor growth. (1)(,) (2)(,) (4) Both vasculogenesis and angiogenesis are driven through signaling by vascular endothelial growth factor (VEGF), and therapeutic

  12. RapGAP9 regulation of the morphogenesis and development in Dictyostelium.

    PubMed

    Mun, Hyemin; Lee, Mi-Rae; Jeon, Taeck J

    2014-04-04

    Recent reports have demonstrated that the importance of Rap1-specific GTPase-activating proteins (GAPs) in the spatial and temporal regulation of Rap1 activity during cell migration and development in Dictyostelium. Here, we identified another putative Rap1 GAP-domain containing protein, showing high sequence homologies with those of human Rap1GAP and Dictyotelium RapGAP3, by bioinformatic search. Loss of RapGAP9 resulted in some defects in morphogenesis and development in Dicytostelium. rapGAP9 null cells were more flattened and spread, and highly multinucleated. Compared to wild-type cells, cells lacking RapGAP9 exhibited increased levels of F-actin and more filopodia. These results suggest that RapGAP9 is involved in the regulation of cytoskeleton reorganization and cytokinesis. rapGAP9 null cells showed a small increase of cell-substratum attachment and slightly lower moving speed and directionality compared to wild-type cells. In addition, the loss of RapGAP9 resulted in an altered morphology of fruiting body with a shorter length of stalk and spore. Identification and characterization of RapGAP9 in this study will provide further insights into the molecular mechanism by which Rap1 regulates cytoskeleton reorganization and morphogenesis in Dictyostelium.

  13. Enhanced cartilage regeneration in MIA/CD-RAP deficient mice.

    PubMed

    Schmid, R; Schiffner, S; Opolka, A; Grässel, S; Schubert, T; Moser, M; Bosserhoff, A-K

    2010-11-11

    Melanoma inhibitory activity/cartilage-derived retinoic acid-sensitive protein (MIA/CD-RAP) is a small soluble protein secreted from chondrocytes. It was identified as the prototype of a family of extracellular proteins adopting an SH3 domain-like fold. In order to study the consequences of MIA/CD-RAP deficiency in detail we used mice with a targeted gene disruption of MIA/CD-RAP (MIA-/-) and analyzed cartilage organisation and differentiation in in vivo and in vitro models. Cartilage formation and regeneration was determined in models for osteoarthritis and fracture healing in vivo, in addition to in vitro studies using mesenchymal stem cells of MIA-/- mice. Interestingly, our data suggest enhanced chondrocytic regeneration in the MIA-/- mice, modulated by enhanced proliferation and delayed differentiation. Expression analysis of cartilage tissue derived from MIA-/- mice revealed strong downregulation of nuclear RNA-binding protein 54-kDa (p54(nrb)), a recently described modulator of Sox9 activity. In this study, we present p54(nrb) as a mediator of MIA/CD-RAP to promote chondrogenesis. Taken together, our data indicate that MIA/CD-RAP is required for differentiation in cartilage potentially by regulating signaling processes during differentiation.

  14. 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.

  15. Rap1-GTP-interacting Adaptor Molecule (RIAM) Protein Controls Invasion and Growth of Melanoma Cells*

    PubMed Central

    Hernández-Varas, Pablo; Coló, Georgina P.; Bartolomé, Ruben A.; Paterson, Andrew; Medraño-Fernández, Iria; Arellano-Sánchez, Nohemí; Cabañas, Carlos; Sánchez-Mateos, Paloma; Lafuente, Esther M.; Boussiotis, Vassiliki A.; Strömblad, Staffan; Teixidó, Joaquin

    2011-01-01

    The Mig-10/RIAM/lamellipodin (MRL) family member Rap1-GTP-interacting adaptor molecule (RIAM) interacts with active Rap1, a small GTPase that is frequently activated in tumors such as melanoma and prostate cancer. We show here that RIAM is expressed in metastatic human melanoma cells and that both RIAM and Rap1 are required for BLM melanoma cell invasion. RIAM silencing in melanoma cells led to inhibition of tumor growth and to delayed metastasis in a severe combined immunodeficiency xenograft model. Defective invasion of RIAM-silenced melanoma cells arose from impairment in persistent cell migration directionality, which was associated with deficient activation of a Vav2-RhoA-ROCK-myosin light chain pathway. Expression of constitutively active Vav2 and RhoA in cells depleted for RIAM partially rescued their invasion, indicating that Vav2 and RhoA mediate RIAM function. These results suggest that inhibition of cell invasion in RIAM-silenced melanoma cells is likely based on altered cell contractility and cell polarization. Furthermore, we show that RIAM depletion reduces β1 integrin-dependent melanoma cell adhesion, which correlates with decreased activation of both Erk1/2 MAPK and phosphatidylinositol 3-kinase, two central molecules controlling cell growth and cell survival. In addition to causing inhibition of cell proliferation, RIAM silencing led to higher susceptibility to cell apoptosis. Together, these data suggest that defective activation of these kinases in RIAM-silenced cells could account for inhibition of melanoma cell growth and that RIAM might contribute to the dissemination of melanoma cells. PMID:21454517

  16. 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.

  17. 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 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...

  18. 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 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...

  19. 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 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...

  20. 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 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...

  1. 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...

  2. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false When do I need a RAP? 270.85 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.85 When do I need a RAP? (a) Whenever you treat, store, or dispose of...

  3. Expression and alternative splicing of N-RAP during mouse skeletal muscle development.

    PubMed

    Lu, Shajia; Borst, Diane E; Horowits, Robert

    2008-12-01

    N-RAP alternative splicing and protein localization were studied in developing skeletal muscle tissue from pre- and postnatal mice and in fusing primary myotubes in culture. Messages encoding N-RAP-s and N-RAP-c, the predominant isoforms of N-RAP detected in adult skeletal muscle and heart, respectively, were present in a 5:1 ratio in skeletal muscle isolated from E16.5 embryos. N-RAP-s mRNA levels increased three-fold over the first 3 weeks of postnatal development, while N-RAP-c mRNA levels remained low. N-RAP alternative splicing during myotube differentiation in culture was similar to the pattern observed in embryonic and neonatal muscle, with N-RAP-s expression increasing and N-RAP-c mRNA levels remaining low. In both developing skeletal muscle and cultured myotubes, N-RAP protein was primarily associated with developing myofibrillar structures containing alpha-actinin, but was not present in mature myofibrils. The results establish that N-RAP-s is the predominant spliced form of N-RAP present throughout skeletal muscle development.

  4. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 27 2014-07-01 2014-07-01 false When do I need a RAP? 270.85 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.85 When do I need a RAP? (a) Whenever you treat, store, or dispose of...

  5. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false When do I need a RAP? 270.85 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.85 When do I need a RAP? (a) Whenever you treat, store, or dispose of...

  6. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 28 2013-07-01 2013-07-01 false When do I need a RAP? 270.85 Section...) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) General Information § 270.85 When do I need a RAP? (a) Whenever you treat, store, or dispose of...

  7. "Fear of a Black Planet": Rap Music and Black Cultural Politics in the 1990s.

    ERIC Educational Resources Information Center

    Rose, Tricia

    1991-01-01

    Explores the exercise of institutional and ideological power over rap music and fans, how artists and fans respond to that context, and the complex relationships between rap's political economy and the sociologically based crime discourse that frames it. Rap's poetic voice is a political expression of the Black experience. (JB)

  8. 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…

  9. In vitro and in vivo study of effects of fermented soybean product (chungkookjang) on NGF secretion ability and NGF receptor signaling pathway

    PubMed Central

    Lee, Young-Ju; Kim, Ji-Eun; Kwak, Moon-Hwa; Go, Jun; Son, Hong-Joo; Kim, Dong-Seob

    2013-01-01

    In order to investigate the effects of a fermented soybean product (Chungkookjang, CKJ) on nerve growth factor (NGF) metabolism, NGF secretion ability and its related signaling pathway were analyzed in B35 neuronal cells and the Tg2576 mouse model of Alzheimer's disease (AD). In B35 cells, the concentration of NGF significantly increased upon treatment with Taegwang (TG)-CKJ and Shinhwa (SH)-CKJ extracts compared with vehicle. Further, a significant increase in PC12 cell length as well as the phsophorylation levels of TrkA and Akt, which are members of a high affinity NGF receptor signaling pathway, were observed after treatment with TG-CKJ and SH-CKJ conditional medium (CM). On the other hand, there was no difference in activation of the NGF receptor p75NTR signaling pathway between vehicle and all CKJ treated groups. In Tg2576 mice showing early stage of AD, the concentrations of NGF in the serum and brain were reduced compared with those in Non-Tg mice. Treatment of Tg2576 mice with SH-CKJ, which contains high concentrations of total flavonoids and phenolic compounds, for 8 weeks dramatically recovered the NGF level to that of Non-Tg mice. Furthermore, the low phosphorylation levels of TrkA and Erk in the NGF receptor TrkA signaling pathway were rapidly recovered to those of Non-Tg mice after SH-CKJ treatment in vehicle treated Tg2576 mice, whereas the phosphorylation level of Akt was maintained at a constant level. These results suggest that CKJ may stimulate NGF secretion ability as well as the NGF receptor TrkA signaling pathway in PC12 cells and Tg2576 mice. PMID:23825484

  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. RAP-1a is the main rhoptry-associated-protein-1 (RAP-1) recognized during infection with Babesia sp. BQ1 (Lintan) (B. motasi-like phylogenetic group), a pathogen of sheep in China.

    PubMed

    Niu, Qingli; Bonsergent, Claire; Rogniaux, Hélène; Guan, Guiquan; Malandrin, Laurence; Moreau, Emmanuelle

    2016-12-15

    Babesia sp. BQ1 (Lintan) is one of the parasites isolated from infected sheep in China that belongs to the B. motasi-like phylogenetic group. The rhoptry-associated-protein 1 (rap-1) locus in this group consists of a complex organization of 12 genes of three main types: 6 rap-1a variants intercalated with 5 identical copies of rap-1b and a single 3' ending rap-1c gene. In the present study, transcription analysis performed by standard RT-PCR demonstrated that the three different rap-1 gene types and the four rap-1a variants were transcribed by the parasite cultivated in vitro. Peptides, specific for each rap-1 type gene, were selected in putative linear B-epitopes and used to raise polyclonal rabbit antisera. Using these sera, the same expression pattern of RAP-1 proteins was found in parasites cultivated in vitro or collected from acute infection whereas only RAP-1a67 was detectable in merozoite extracts. However, ELISA performed with recombinant RAP-1a67, RAP-1b or RAP-1c and sera from infected sheep demonstrated that RAP-1a67 is the main RAP-1 recognized during infection, even if some infected sheep also recognized RAP-1b and/or RAP-1c.

  12. Comparison of RapID Yeast Plus System with API 20C System for Identification of Common, New, and Emerging Yeast Pathogens

    PubMed Central

    Espinel-Ingroff, A.; Stockman, L.; Roberts, G.; Pincus, D.; Pollack, J.; Marler, J.

    1998-01-01

    The ability to identify yeast isolates by the new enzymatic RapID Yeast Plus System was compared to the ability to identify yeast isolates by the API 20C system. A total of 447 yeast isolates representing Blastoschizomyces capitatus, 17 Candida spp., 5 Cryptococcus spp., Geotrichum spp., 2 Hanseniaspora spp., Hansenula anomala, Hansenula wingei, 3 Rhodotorula spp., Saccharomyces cerevisiae, Sporobolomyces salmonicolor, Trichosporon beigelii, and 2 Prototheca spp. were evaluated. Also, five quality control strains (Candida spp. and Cryptococcus laurentii) with well-documented reactivities by the RapID Yeast Plus System were used. Each isolate was evaluated by both methods with a 48-h culture grown at 30°C on Sabouraud dextrose agar (Emmons modification) by following the recommendations of the manufacturers. The RapID Yeast Plus System enzymatic reactions were read after 4 h of incubation, and the API 20C carbohydrate assimilation identification profiles were obtained after 72 h of incubation. There was good (95.7%) agreement between the identifications obtained by the two methods with the eight common Candida spp. and with Cryptococcus neoformans. The agreement was lower when the emerging Candida spp. and other yeast-like pathogens were tested (79.1 and 75.2%, respectively). These preliminary data suggest the potential utility of the RapID Yeast Plus System for use in the clinical laboratory for the rapid identification of common yeast pathogens as well as certain new and emerging species. PMID:9542903

  13. Comparison of RapID yeast plus system with API 20C system for identification of common, new, and emerging yeast pathogens.

    PubMed

    Espinel-Ingroff, A; Stockman, L; Roberts, G; Pincus, D; Pollack, J; Marler, J

    1998-04-01

    The ability to identify yeast isolates by the new enzymatic RapID Yeast Plus System was compared to the ability to identify yeast isolates by the API 20C system. A total of 447 yeast isolates representing Blastoschizomyces capitatus, 17 Candida spp., 5 Cryptococcus spp., Geotrichum spp., 2 Hanseniaspora spp., Hansenula anomala, Hansenula wingei, 3 Rhodotorula spp., Saccharomyces cerevisiae, Sporobolomyces salmonicolor, Trichosporon beigelii, and 2 Prototheca spp. were evaluated. Also, five quality control strains (Candida spp. and Cryptococcus laurentii) with well-documented reactivities by the RapID Yeast Plus System were used. Each isolate was evaluated by both methods with a 48-h culture grown at 30 degrees C on Sabouraud dextrose agar (Emmons modification) by following the recommendations of the manufacturers. The RapID Yeast Plus System enzymatic reactions were read after 4 h of incubation, and the API 20C carbohydrate assimilation identification profiles were obtained after 72 h of incubation. There was good (95.7%) agreement between the identifications obtained by the two methods with the eight common Candida spp. and with Cryptococcus neoformans. The agreement was lower when the emerging Candida spp. and other yeast-like pathogens were tested (79.1 and 75.2%, respectively). These preliminary data suggest the potential utility of the RapID Yeast Plus System for use in the clinical laboratory for the rapid identification of common yeast pathogens as well as certain new and emerging species.

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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,...

  20. 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

    ... 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...

  1. 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,...

  2. 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...

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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 §...

  11. 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...

  12. 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...

  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 270.175 - 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 270.90 - 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 §...

  16. 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...

  17. 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,...

  18. 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,...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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 §...

  5. 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...

  6. 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 §...

  7. 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...

  8. 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...

  9. 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...

  10. 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 §...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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,...

  16. 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...

  17. 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...

  18. Characterize RAP80, a Potential Tumor Suppressor Gene

    DTIC Science & Technology

    2009-04-01

    Fanconi anemia complementation group D2 ( FANCD2 ). However, RAP80 foci still form normally after irradiation in FANCD2 -deficient cells (fig. S7...encoding HSJ1A and Ubi-GST, respectively; and A. D’Andrea for providing FANCD2 -deficient and FANCD2 -reconstituted cells. This work was supported by grants

  19. 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…

  20. 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,…

  1. 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…

  2. 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…

  3. 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.

  4. 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…

  5. Teaching Literacy as Rap at Southeast Community College.

    ERIC Educational Resources Information Center

    Sundeen, Jim

    2003-01-01

    Describes how the author became critically aware of the dynamics of literacy and race in a composition classroom. Introduces his students to rap music as a legitimate literacy and a type of literature in its own right. Describes the lesson plan he used in the project and explains some of the theory that inspired his classroom inquiry. (SG)

  6. MAGI-1 Interacts with Nephrin to Maintain Slit Diaphragm Structure through Enhanced Rap1 Activation in Podocytes*

    PubMed Central

    Ni, Jie; Bao, Sujin; Johnson, Ruth I.; Zhu, Bingbing; Li, Jianhua; Vadaparampil, Justin; Smith, Christopher M.; Campbell, Kirk N; Grahammer, Florian; Huber, Tobias B.; He, John C.; D'Agati, Vivette D.; Chan, Andrew; Kaufman, Lewis

    2016-01-01

    MAGI-1 is a multidomain cytosolic scaffolding protein that in the kidney is specifically located at the podocyte slit diaphragm, a specialized junction that is universally injured in proteinuric diseases. There it interacts with several essential molecules, including nephrin and neph1, which are required for slit diaphragm formation and as an intracellular signaling hub. Here, we show that diminished MAGI-1 expression in cultured podocytes reduced nephrin and neph1 membrane localization and weakened tight junction integrity. Global magi1 knock-out mice, however, demonstrated normal glomerular histology and function into adulthood. We hypothesized that a second mild but complementary genetic insult might induce glomerular disease susceptibility in these mice. To identify such a gene, we utilized the developing fly eye to test for functional complementation between MAGI and its binding partners. In this way, we identified diminished expression of fly Hibris (nephrin) or Roughest (neph1) as dramatically exacerbating the effects of MAGI depletion. Indeed, when these combinations were studied in mice, the addition of nephrin, but not neph1, heterozygosity to homozygous deletion of MAGI-1 resulted in spontaneous glomerulosclerosis. In cultured podocytes, MAGI-1 depletion reduced intercellular contact-induced Rap1 activation, a pathway critical for proper podocyte function. Similarly, magi1 knock-out mice showed diminished glomerular Rap1 activation, an effect dramatically enhanced by concomitant nephrin haploinsufficiency. Finally, combined overexpression of MAGI-1 and nephrin increased Rap1 activation, but not when substituting a mutant MAGI-1 that cannot bind nephrin. We conclude that the interaction between nephrin and MAGI-1 regulates Rap1 activation in podocytes to maintain long term slit diaphragm structure. PMID:27707879

  7. Myofibril assembly visualized by imaging N-RAP, alpha-actinin, and actin in living cardiomyocytes.

    PubMed

    Manisastry, Shyam M; Zaal, Kristien J M; Horowits, Robert

    2009-07-15

    N-RAP is a striated muscle-specific scaffolding protein that organizes alpha-actinin and actin into symmetrical I-Z-I structures in developing myofibrils. Here we determined the order of events during myofibril assembly through time-lapse confocal microscopy of cultured embryonic chick cardiomyocytes coexpressing fluorescently tagged N-RAP and either alpha-actinin or actin. During de novo myofibril assembly, N-RAP assembled in fibrillar structures within the cell, with dots of alpha-actinin subsequently organizing along these structures. The initial fibrillar structures were reminiscent of actin fibrils, and coassembly of N-RAP and actin into newly formed fibrils supported this. The alpha-actinin dots subsequently broadened to Z-lines that were wider than the underlying N-RAP fibril, and N-RAP fluorescence intensity decreased. FRAP experiments showed that most of the alpha-actinin dynamically exchanged during all stages of myofibril assembly. In contrast, less than 20% of the N-RAP in premyofibrils was exchanged during 10-20 min after photobleaching, but this value increased to 70% during myofibril maturation. The results show that N-RAP assembles into an actin containing scaffold before alpha-actinin recruitment; that the N-RAP scaffold is much more stable than the assembling structural components; that N-RAP dynamics increase as assembly progresses; and that N-RAP leaves the structure after assembly is complete.

  8. 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.

  9. 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.

  10. Yeast telomere repeat sequence (TRS) improves circular plasmid segregation, and TRS plasmid segregation involves the RAP1 gene product.

    PubMed Central

    Longtine, M S; Enomoto, S; Finstad, S L; Berman, J

    1992-01-01

    Telomere repeat sequences (TRSs) can dramatically improve the segregation of unstable circular autonomously replicating sequence (ARS) plasmids in Saccharomyces cerevisiae. Deletion analysis demonstrated that yeast TRSs, which conform to the general sequence (C(1-3)A)n, are able to stabilize circular ARS plasmids. A number of TRS clones of different primary sequence and C(1-3)A tract length confer the plasmid stabilization phenotype. TRS sequences do not appear to improve plasmid replication efficiency, as determined by plasmid copy number analysis and functional assays for ARS activity. Pedigree analysis confirms that TRS-containing plasmids are missegregated at low frequency and that missegregated TRS-containing plasmids, like ARS plasmids, are preferentially retained by the mother cell. Plasmids stabilized by TRSs have properties that distinguish them from centromere-containing plasmids and 2 microns-based recombinant plasmids. Linear ARS plasmids, which include two TRS tracts at their termini, segregate inefficiently, while circular plasmids with one or two TRS tracts segregate efficiently, suggesting that plasmid topology or TRS accessibility interferes with TRS segregation function on linear plasmids. In strains carrying the temperature-sensitive mutant alleles rap1grc4 and rap1-5, TRS plasmids are not stable at the semipermissive temperature, suggesting that RAP1 protein is involved in TRS plasmid stability. In Schizosaccharomyces pombe, an ARS plasmid was stabilized by the addition of S. pombe telomere sequence, suggesting that the ability to improve the segregation of ARS plasmids is a general property of telomere repeats. PMID:1569937

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. Interleukin-18 augments growth ability of primary human melanocytes by PTEN inactivation through the AKT/NF-κB pathway.

    PubMed

    Zhou, Jia; Shang, Jing; Song, Jing; Ping, Fengfeng

    2013-02-01

    Normal human skin relies on melanocytes to provide photoprotection and thermoregulation by producing melanin. The growth and behavior of melanocytes are controlled by many factors. Interleukin-18 (IL-18) is expressed in both immune and non-immune cells and participates in the adjustment of multitude cellular functions. Nonetheless, the regulative roles of IL-18 in melanogenesis and growth of melanocytes have not been explored. The present study was conducted to investigate the effects of IL-18 on melanocytes and elucidate the underlying mechanisms. We proved that IL-18 increased the tyrosinase activity and melanin content in normal human foreskin-derived epidermal melanocytes (NHEM). Treatment with IL-18 (20 ng/ml) enhanced the expression of c-Kit, microphtalmia-associated transcription factor (MITF) and its downstream tyrosinase-related protein 1 (TRP-1), and TRP-2. In addition, IL-18 induced NHEM migration at concentration of 20 ng/ml. These results indicated a promotive action of IL-18 on melanogenesis in NHEM. Our data revealed that IL-18 stimulated ERK1/2 and NF-κB activation, improved p-Akt, p70 S6K and anti-apoptotic Bcl-2 levels, and deactivated phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in NHEM. Besides, IL-18 increased level of PTEN phosphorylation to protect NHEM from damage induced by H(2)O(2). These results in vitro showed the accommodation of IL-18 in melanocytes growth. Therefore, we suggested an important regulating action of IL-18 to melanogenesis and cell growth ability of skin melanocytes.

  17. Sevoflurane represses the self-renewal ability by regulating miR-7a,7b/Klf4 signalling pathway in mouse embryonic stem cells.

    PubMed

    Wang, Qimin; Li, Guifeng; Li, Baolin; Chen, Qiu; Lv, Dongdong; Liu, Jiaying; Ma, Jieyu; Sun, Nai; Yang, Longqiu; Fei, Xuejie; Song, Qiong

    2016-10-01

    Sevoflurane is a frequently-used clinical inhalational anaesthetic and can cause toxicity to embryos during foetal development. Embryonic stem cells (ESCs) are derived from the inner cell mass of blastospheres and can be used as a useful model of early development. Here, we found that sevoflurane significantly influenced self-renewal ability of mESCs on stemness maintenance and cell proliferation. The cell cycle was arrested via G1 phase delay. We further found that sevoflurane upregulated expression of miR-7a,7b to repress self-renewal. Next we performed rescue experiments and found that after adding miR-7a,7b inhibitor into mESCs treated with sevoflurane, its influence on self-renewal could be blocked. Further we identified stemness factor Klf4 as the direct target of miR-7a,7b. Overexpression of Klf4 restored self-renewal ability repressed by miR-7a,7b or sevoflurane. In this work, we determined that sevoflurane repressed self-renewal ability by regulating the miR-7a,7b/Klf4 signalling pathway in mESCs. Our study demonstrated molecular mechanism underlying the side effects of sevoflurane during early development, laying the foundation for studies on safe usage of inhalational anaesthetic during non-obstetric surgery.

  18. 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.

  19. “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

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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

  6. Spa-1 (Sipa1) and Rap signaling in leukemia and cancer metastasis.

    PubMed

    Minato, Nagahiro; Hattori, Masakazu

    2009-01-01

    Although Rap GTPases of the Ras family remained enigmatic for years, extensive studies in this decade have revealed diverse functions of Rap in the control of cell proliferation, differentiation, survival, adhesion, and movement. With the use of genetic engineering strategies, we have uncovered essential roles of Rap signaling in normal lymphohematopoietic cell development as well as its crucial involvement in the development of a wide spectrum of leukemia in manners highly dependent on the contexts of cell lineages. Incidentally, recent results also indicate an important role of Spa-1, a Rap GTPase-activating protein, in invasion and metastasis in human cancers. While it is unlikely that Rap can function as a classic oncogene by itself, like Ras, emerging findings unveil crucial involvements of Rap GTPases in the distinct aspects of malignancy, including leukemia genesis and cancer metastasis.

  7. GKN2 increases apoptosis, reduces the proliferation and invasion ability of gastric cancer cells through down-regulating the JAK/STAT signaling pathway

    PubMed Central

    Ouyang, Jun; Pan, Xiaohui; Lin, Hui; Hu, Zecheng; Xiao, Ping; Hu, Haobin

    2017-01-01

    Objectives: To investigate the effect of gastric motility protein 2 (GKN2) on the proliferation, apoptosis and invasion of gastric cancer cell and on the JAK/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods: Expression of GKN2 was qualified using Western blot analysis in four gastric cancer cell lines and immortalized human gastric mucosal epithelial cell line GES-1. The cells were then transfected with pcDNA3.1-GKN2 and control vector using Lipofectaminetm2000 and assayed for viability, apoptosis, cell cycle changes, invasion ability as well as expression of cell cycle protein D1 (Cylin D1), Bcl-2, Bax, matrix metalloproteinase 2 (MMP2), MMP9, JAK2 and p-STAT3. Results: Western blot analyses showed that the expression of GKN2 was significantly lower in 4 gastric cancer cell lines (BGC-823, SGC-7901, AGS and MKN-45) than in GES-1. Of them, SGC-7901 had the lowest expression. The line was chosen for subsequent transfection experiments. Compared with control (transfection with empty vector), pcDNA3.1-GKN2-transfected cells had significantly more GKN2 protein and mRNA, decreased cell viability, increased apoptosis, more cells arrested at G1 phase and reduced invasiveness. Expression analyses showed that expression of Cyclin D1, Bcl-2, MMP2, MMP9, JAK2 and STAT3 was significantly down-regulated, while Bax was significantly up-regulated. Conclusion: Over-expression of GKN2 can increase apoptosis, reduce proliferation and invasion ability of gastric cancer cells as a result of down-regulated JAK2/STAT3 signaling pathway. PMID:28337309

  8. Teratogen-induced distortions in the classical NF-{kappa}B activation pathway: Correlation with the ability of embryos to survive teratogenic stress

    SciTech Connect

    Molotski, Natali; Savion, Shoshana; Gerchikov, Natalie; Fein, Amos; Toder, Vladimir; Torchinsky, Arkady

    2008-06-01

    Studies with diverse teratogens implicated the transcription factor NF-{kappa}B in mechanisms determining teratological susceptibility of embryos. Here, a teratogen such as cyclophosphamide (CP) was used to test whether teratogenic insult alters the classical NF-{kappa}B activation pathway, and how these alterations correlate with the ability of mouse embryos to resist the teratogen-induced process of maldevelopment. We observed that embryos tested 24 h after the exposure of females to 40 mg/kg CP exhibited a dramatic decrease in the level of NF-{kappa}B (p65 subunit)-DNA binding, I{kappa}B kinase beta (IKK{beta}) activity, expression of p65 and IKK{beta} proteins, as well as NF-{kappa}B inhibitory proteins (I{kappa}Bs) such as I{kappa}B{alpha}, I{kappa}B{beta}, and I{kappa}B{epsilon}, and died within the next 24 h. Embryos of females exposed to 15 mg/kg CP exhibited only a decrease in NF-{kappa}B-DNA binding and IKK{beta} activity at 24 h. However, at 48 h, a more prominent decrease in NF-{kappa}B activity was observed, accompanied by a decreased expression of p65 and IKK{beta} proteins. These embryos died within the next 24 h. After treatment with 10 mg/kg CP, embryos survived until the end of the antenatal period of development, demonstrating a transient decrease in NF-{kappa}B-DNA binding activity and no alterations in NF-{kappa}B signaling. These results suggest that the classical NF-{kappa}B activation pathway may be among targets that teratogens engage to initiate abnormal development. Besides, the observation that embryos destined to be dead exhibited a dramatically decreased rate of cell proliferation suggests a pathway, whereby teratogen-induced alterations in NF-{kappa}B signaling may culminate in such a final effect as embryonic death.

  9. B cell receptor-induced phosphorylation of Pyk2 and focal adhesion kinase involves integrins and the Rap GTPases and is required for B cell spreading.

    PubMed

    Tse, Kathy W K; Dang-Lawson, May; Lee, Rosaline L; Vong, Doris; Bulic, Anica; Buckbinder, Leonard; Gold, Michael R

    2009-08-21

    Signaling by the B cell receptor (BCR) promotes integrin-mediated adhesion and cytoskeletal reorganization. This results in B cell spreading, which enhances the ability of B cells to bind antigens and become activated. Proline-rich tyrosine kinase (Pyk2) and focal adhesion kinase (FAK) are related cytoplasmic tyrosine kinases that regulate cell adhesion, cell morphology, and cell migration. In this report we show that BCR signaling and integrin signaling collaborate to induce the phosphorylation of Pyk2 and FAK on key tyrosine residues, a modification that increases the kinase activity of Pyk2 and FAK. Activation of the Rap GTPases is critical for BCR-induced integrin activation as well as for BCR- and integrin-induced reorganization of the actin cytoskeleton. We now show that Rap activation is essential for BCR-induced phosphorylation of Pyk2 and for integrin-induced phosphorylation of Pyk2 and FAK. Moreover Rap-dependent phosphorylation of Pyk2 and FAK required an intact actin cytoskeleton as well as actin dynamics, suggesting that Rap regulates Pyk2 and FAK via its effects on the actin cytoskeleton. Importantly B cell spreading induced by BCR/integrin co-stimulation or by integrin engagement was inhibited by short hairpin RNA-mediated knockdown of either Pyk2 or FAK expression and by treatment with PF-431396, a chemical inhibitor that blocks the kinase activities of both Pyk2 and FAK. Thus Pyk2 and FAK are downstream targets of the Rap GTPases that play a key role in regulating B cell morphology.

  10. 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.

  11. 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.

  12. 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.

  13. Enhancement of the proline and nitric oxide synthetic pathway improves fermentation ability under multiple baking-associated stress conditions in industrial baker's yeast

    PubMed Central

    2012-01-01

    plays an important role in baking-associated stress tolerance. Conclusions In this work, we clarified the importance of Put1- and Mpr1-mediated NO generation from proline to the baking-associated stress tolerance in industrial baker's yeast. We also demonstrated that baker's yeast that enhances the proline and NO synthetic pathway by expressing the Pro1-I150T and Mpr1-F65L variants showed improved fermentation ability under multiple baking-associated stress conditions. From a biotechnological perspective, the enhancement of proline and NO synthesis could be promising for breeding novel baker's yeast strains. PMID:22462683

  14. 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.

  15. 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

  16. 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.

  17. HS1 deficiency impairs neutrophil recruitment in vivo and activation of the small GTPases Rac1 and Rap1.

    PubMed

    Latasiewicz, Joanna; Artz, Annette; Jing, Ding; Blanco, Mariana Pacheco; Currie, Silke M; Avila, Martha Velázquez; Schnoor, Michael; Vestweber, Dietmar

    2017-01-25

    Neutrophil extravasation is a critical step of the innate immune system's response to inflammation. This multistep process is tightly regulated by adhesion and signaling molecules in the endothelium and neutrophils. Activation of the β2 integrin LFA-1 is critical for adhesion of leukocytes to postcapillary venules. This step requires coordinated activation of signaling pathways in chemokine-stimulated neutrophils, including GTPase activation and cytoskeletal remodeling, leading to conformational changes in LFA-1. Hematopoietic cell-specific lyn substrate 1 (HS1) is a cortactin-related and leukocyte-specific actin-binding protein (ABP) that regulates several processes in various immune cells. It has been shown in vitro that HS1 is important for neutrophil chemotaxis and transendothelial migration of NK cells, but its role in neutrophil extravasation in vivo has not been investigated yet. Intravital microscopy of CXCL1-stimulated cremaster venules revealed an increased rolling velocity and reduced neutrophil adhesion and transmigration in HS1 knockout (KO) mice. CXCL1-induced rapid neutrophil arrest in vivo and adhesion under flow conditions in vitro were also reduced significantly. Whereas random motility of neutrophils was unaffected, chemotaxis toward a CXCL1 gradient was reduced in the absence of HS1. Further analysis of the underlying mechanisms demonstrated that HS1 controls CXCL1-induced activation of the small GTPases Ras-related C3 botulinum toxin substrate 1 (Rac1) and Ras-related protein 1 (Rap1), thus supporting LFA-1-mediated neutrophil adhesion. Importantly, with the use of Rac1 KO neutrophils, we could show that Rac1 acts upstream of Rap1. Our results establish HS1 as an important regulator of proper Rac1 and Rap1 activation and neutrophil extravasation.

  18. Requirement for Plk2 in orchestrated ras and rap signaling, homeostatic structural plasticity, and memory.

    PubMed

    Lee, Kea Joo; Lee, Yeunkum; Rozeboom, Aaron; Lee, Ji-Yun; Udagawa, Noriko; Hoe, Hyang-Sook; Pak, Daniel T S

    2011-03-10

    Ras and Rap small GTPases are important for synaptic plasticity and memory. However, their roles in homeostatic plasticity are unknown. Here, we report that polo-like kinase 2 (Plk2), a homeostatic suppressor of overexcitation, governs the activity of Ras and Rap via coordination of their regulatory proteins. Plk2 directs elimination of Ras activator RasGRF1 and Rap inhibitor SPAR via phosphorylation-dependent ubiquitin-proteasome degradation. Conversely, Plk2 phosphorylation stimulates Ras inhibitor SynGAP and Rap activator PDZGEF1. These Ras/Rap regulators perform complementary functions to downregulate dendritic spines and AMPA receptors following elevated activity, and their collective regulation by Plk2 profoundly stimulates Rap and suppresses Ras. Furthermore, perturbation of Plk2 disrupts Ras and Rap signaling, prevents homeostatic shrinkage and loss of dendritic spines, and impairs proper memory formation. Our study demonstrates a critical role of Plk2 in the synchronized tuning of Ras and Rap and underscores the functional importance of this regulation in homeostatic synaptic plasticity.

  19. CDK5RAP3 is a novel repressor of p14ARF in hepatocellular carcinoma cells.

    PubMed

    Mak, Grace Wing-Yan; Lai, Wai-Lung; Zhou, Yuan; Li, Mingtao; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2012-01-01

    CDK5 regulatory subunit associated protein 3 (CDK5RAP3) is a novel activator of PAK4 and processes important pro-metastatic function in hepatocarcinogenesis. However, it remains unclear if there are other mechanisms by which CDK5RAP3 promotes HCC metastasis. Here, we showed that in CDK5RAP3 stable knockdown SMMC-7721 HCC cells, p14(ARF) tumor suppressor was upregulated at protein and mRNA levels, and ectopic expression of CDK5RAP3 was found to repress the transcription of p14(ARF). Using chromatin immunoprecipitation assay, we demonstrated that CDK5RAP3 bound to p14(ARF) promoter in vivo. Furthermore, knockdown of p14(ARF) in CDK5RAP3 stable knockdown HCC cells reversed the suppression of HCC cell invasiveness mediated by knockdown of CDK5RAP3. Taken together, our findings provide the new evidence that overexpression of CDK5RAP3 promotes HCC metastasis via downregulation of p14(ARF).

  20. CDK5RAP3 Is a Novel Repressor of p14ARF in Hepatocellular Carcinoma Cells

    PubMed Central

    Mak, Grace Wing-Yan; Li, Mingtao; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2012-01-01

    CDK5 regulatory subunit associated protein 3 (CDK5RAP3) is a novel activator of PAK4 and processes important pro-metastatic function in hepatocarcinogenesis. However, it remains unclear if there are other mechanisms by which CDK5RAP3 promotes HCC metastasis. Here, we showed that in CDK5RAP3 stable knockdown SMMC-7721 HCC cells, p14ARF tumor suppressor was upregulated at protein and mRNA levels, and ectopic expression of CDK5RAP3 was found to repress the transcription of p14ARF. Using chromatin immunoprecipitation assay, we demonstrated that CDK5RAP3 bound to p14ARF promoter in vivo. Furthermore, knockdown of p14ARF in CDK5RAP3 stable knockdown HCC cells reversed the suppression of HCC cell invasiveness mediated by knockdown of CDK5RAP3. Taken together, our findings provide the new evidence that overexpression of CDK5RAP3 promotes HCC metastasis via downregulation of p14ARF. PMID:22860085

  1. 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…

  2. 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…

  3. 40 CFR 270.85 - When do I need a RAP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false When do I need a RAP? 270.85 Section 270.85 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED...) General Information § 270.85 When do I need a RAP? (a) Whenever you treat, store, or dispose of...

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

    PubMed

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

    2015-10-01

    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.

  5. The Boisterous Beauty of the Urban Bard: Houston Baker Crusades for the Cultural Relevance of Rap.

    ERIC Educational Resources Information Center

    Wiley, Ed, III

    1993-01-01

    Examines Houston Baker's argument for rap music's place in academe. Baker suggests rap as being one of the most creative and productive forms of cultural expression to come along in some time warranting the same kind of scholarly attention afforded the works of history's most noted philosophers and poets. (GLR)

  6. Black Studies, Rap and the Academy. Black Literature and Culture Series.

    ERIC Educational Resources Information Center

    Baker, Houston A., Jr.

    The relationships among Black Studies as an intellectual discipline and rap music are explored. It is argued that black urban culture has provided much of the impetus for Black Studies, and that the academy and those involved in the black studies discipline should feel a responsibility to take rap music seriously as the expression of urban youth,…

  7. 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…

  8. A Receptor-associated Protein/Phosphatidylinositol 3-Kinase Pathway Controls Pseudopod Formation

    PubMed Central

    Kortholt, Arjan; Bolourani, Parvin; Rehmann, Holger; Keizer-Gunnink, Ineke; Weeks, Gerald; Wittinghofer, Alfred

    2010-01-01

    GbpD, a Dictyostelium discoideum guanine exchange factor specific for Rap1, has been implicated in adhesion, cell polarity, and chemotaxis. Cells overexpressing GbpD are flat, exhibit strongly increased cell-substrate attachment, and extend many bifurcated and lateral pseudopodia. Phg2, a serine/threonine-specific kinase, mediates Rap1-regulated cell-substrate adhesion, but not cell polarity or chemotaxis. In this study we demonstrate that overexpression of GbpD in pi3k1/2-null cells does not induce the adhesion and cell morphology phenotype. Furthermore we show that Rap1 directly binds to the Ras binding domain of PI3K, and overexpression of GbpD leads to strongly enhanced PIP3 levels. Consistently, upon overexpression of the PIP3-degradating enzyme PTEN in GbpD-overexpressing cells, the strong adhesion and cell morphology phenotype is largely lost. These results indicate that a GbpD/Rap/PI3K pathway helps control pseudopod formation and cell polarity. As in Rap-regulated pseudopod formation in Dictyostelium, mammalian Rap and PI3K are essential for determining neuronal polarity, suggesting that the Rap/PI3K pathway is a conserved module regulating the establishment of cell polarity. PMID:20089846

  9. Cantharidin and norcantharidin inhibit the ability of MCF-7 cells to adhere to platelets via protein kinase C pathway-dependent downregulation of α2 integrin.

    PubMed

    Shou, Liu-Mei; Zhang, Qiong-Yan; Li, Wei; Xie, Xin; Chen, Kai; Lian, Lian; Li, Zhen-Yu; Gong, Fei-Ran; Dai, Ke-Sheng; Mao, Yi-Xiang; Tao, Min

    2013-09-01

    Cancer metastasis is a highly coordinated and dynamic multistep process in which cancer cells interact with a variety of host cells. Morphological studies have documented the association of circulating tumor cells with host platelets, where a surface coating of platelets protects tumor cells from mechanical trauma and the immune system. Cantharidin is an active constituent of mylabris, a traditional Chinese medicine. Cantharidin and norcantharidin are potent protein phosphatase 2A (PP2A) inhibitors that exhibit in vitro and in vivo antitumor activity against several types of cancer, including breast cancer. We investigated whether cantharidin and norcantharidin could repress the ability of MCF-7 breast cancer cells to adhere to platelets. Using MTT, clone formation, apoptosis, adhesion and wound-healing assays, we found that cantharidin and norcantharidin induced apoptosis and repressed MCF-7 cell growth, adhesion and migration. Moreover, we developed a flow cytometry-based analysis of tumor cell adhesion to platelets. We proved that cantharidin and norcantharidin repressed MCF-7 cell adhesion to platelets through downregulation of α2 integrin, an adhesion molecule present on the surface of cancer cells. The repression of α2 integrin expression was found to be executed through the protein kinase C pathway, the activation of which could have been due to PP2A inhibition.

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. Platinum(IV) complex LA-12 exerts higher ability than cisplatin to enhance TRAIL-induced cancer cell apoptosis via stimulation of mitochondrial pathway.

    PubMed

    Jelínková, Iva; Šafaříková, Barbora; Vondálová Blanářová, Olga; Skender, Belma; Hofmanová, Jiřina; Sova, Petr; Moyer, Mary Pat; Kozubík, Alois; Kolář, Zdeněk; Ehrmann, Jiří; Hyršlová Vaculová, Alena

    2014-12-01

    In search for novel strategies in colon cancer treatment, we investigated the unique ability of platinum(IV) complex LA-12 to efficiently enhance the killing effects of tumor necrosis factor-related apoptosis inducing ligand (TRAIL), and compared it with the sensitizing action of cisplatin. We provide the first evidence that LA-12 primes human colon cancer cells for TRAIL-induced cytotoxicity by p53-independent activation of the mitochondrial apoptotic pathway. The cooperative action of LA-12 and TRAIL was associated with stimulation of Bax/Bak activation, drop of mitochondrial membrane potential, caspase-9 activation, and a shift of the balance among Bcl-2 family proteins in favor of the pro-apoptotic members. In contrast to cisplatin, LA-12 was a potent inducer of ERK-mediated Noxa and BimL protein upregulation, and more effectively enhanced TRAIL-induced apoptosis in the absence of Bax. The cooperative action of LA-12 and TRAIL was augmented following the siRNA-mediated silencing of Mcl-1 in both Bax proficient/deficient cells. We newly demonstrated that LA-12 induced ERK-mediated c-Myc upregulation, and proved that c-Myc silencing inhibited the mitochondrial activation and apoptosis in colon cancer cells treated with LA-12 and TRAIL. The LA-12-mediated sensitization to TRAIL-induced apoptosis was demonstrated in several colon cancer cell lines, further underscoring the general relevance of our findings. The selective action of LA-12 was documented by preferential priming of cancer but not normal colon cancer cells to TRAIL killing effects. Our work highlights the promising potential of LA-12 over cisplatin to enhance the colon cancer cell sensitivity to TRAIL-induced apoptosis, and provides new mechanistic insights into their cooperative action.

  16. 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...

  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, 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...

  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, 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)...

  19. 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...

  20. 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...

  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, 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...

  2. 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)...

  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, 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...

  4. 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)...

  5. 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...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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)...

  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, 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...

  12. 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.

  13. 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.

  14. KIF14 negatively regulates Rap1a–Radil signaling during breast cancer progression

    PubMed Central

    Ahmed, Syed M.; Thériault, Brigitte L.; Uppalapati, Maruti; Chiu, Catherine W.N.; Gallie, Brenda L.; Sidhu, Sachdev S.

    2012-01-01

    The small GTPase Rap1 regulates inside-out integrin activation and thereby influences cell adhesion, migration, and polarity. Several Rap1 effectors have been described to mediate the cellular effects of Rap1 in a context-dependent manner. Radil is emerging as an important Rap effector implicated in cell spreading and migration, but the molecular mechanisms underlying its functions are unclear. We report here that the kinesin KIF14 associates with the PDZ domain of Radil and negatively regulates Rap1-mediated inside-out integrin activation by tethering Radil on microtubules. The depletion of KIF14 led to increased cell spreading, altered focal adhesion dynamics, and inhibition of cell migration and invasion. We also show that Radil is important for breast cancer cell proliferation and for metastasis in mice. Our findings provide evidence that the concurrent up-regulation of Rap1 activity and increased KIF14 levels in several cancers is needed to reach optimal levels of Rap1–Radil signaling, integrin activation, and cell–matrix adhesiveness required for tumor progression. PMID:23209302

  15. Rap2b, a novel p53 target, regulates p53-mediated pro-survival function

    PubMed Central

    Zhang, Xinyue; He, Yunlong; Lee, Kyoung-Hwa; Dubois, Wendy; Li, Ziqing; Wu, Xiaolin; Kovalchuk, Alexander; Zhang, Weimin; Huang, Jing

    2013-01-01

    The tumor suppressor p53 is a critical regulator of apoptosis and cell cycle arrest/pro-survival. Upon DNA damage, p53 evokes both cell cycle arrest/pro-survival and apoptosis transcriptional programs. The ultimate cellular outcome depends on the balance of these two programs. However, the p53 downstream targets that mediate this cell fate decision remain to be identified. Using an integrative genomic approach, we identify Rap2b as a conserved p53-activated gene that counters p53-mediated apoptosis after DNA damage. Upon DNA damage, p53 directly binds to the promoter of Rap2b and activates its transcription. The reduction of Rap2b levels by small interference RNA sensitizes cells to DNA damage-induced apoptosis in a p53-dependent manner. Consistent with its pro-survival function, analysis of cancer genomic data reveals that Rap2b is overexpressed in many types of tumors. Anchorage-independent growth assays show that Rap2b has only weak transformation activity, suggesting that it is not an oncogene by itself. Together, our results identify Rap2b as a new player in the pro-survival program conducted by p53 and raise the possibility that targeting Rap2b could sensitize tumor cells to apoptosis in response to DNA damage. PMID:23535297

  16. Defective angiogenesis, endothelial migration, proliferation, and MAPK signaling in Rap1b-deficient mice

    PubMed Central

    Kraus, Anna E.; Gale, Daniel; White, Gilbert C.; VanSluys, Jillian

    2008-01-01

    Angiogenesis is the main mechanism of vascular remodeling during late development and, after birth, in wound healing. Perturbations of angiogenesis occur in cancer, diabetes, ischemia, and inflammation. While much progress has been made in identifying factors that control angiogenesis, the understanding of the precise molecular mechanisms involved is incomplete. Here we identify a small GTPase, Rap1b, as a positive regulator of angiogenesis. Rap1b-deficient mice had a decreased level of Matrigel plug and neonatal retinal neovascularization, and aortas isolated from Rap1b-deficient animals had a reduced microvessel sprouting response to 2 major physiological regulators of angiogenesis: vascular endothelial growth factor (VEGF) and basic fibroblasts growth factor (bFGF), indicating an intrinsic defect in endothelial cells. Proliferation of retinal endothelial cells in situ and in vitro migration of lung endothelial cells isolated from Rap1b-deficient mice were inhibited. At the molecular level, activation of 2 MAP kinases, p38 MAPK and p42/44 ERK, important regulators of endothelial migration and proliferation, was decreased in Rap1b-deficient endothelial cells in response to VEGF stimulation. These studies provide evidence that Rap1b is required for normal angiogenesis and reveal a novel role of Rap1 in regulation of proangiogenic signaling in endothelial cells. PMID:17993608

  17. Rap1 and its effector RIAM are required for lymphocyte trafficking.

    PubMed

    Su, Wenjuan; Wynne, Joseph; Pinheiro, Elaine M; Strazza, Marianne; Mor, Adam; Montenont, Emilie; Berger, Jeffrey; Paul, David S; Bergmeier, Wolfgang; Gertler, Frank B; Philips, Mark R

    2015-12-17

    Regulation of integrins is critical for lymphocyte adhesion to endothelium and trafficking through secondary lymphoid organs. Inside-out signaling to integrins is mediated by the small GTPase Rap1. Two effectors of Rap1 regulate integrins, RapL and Rap1 interacting adaptor molecule (RIAM). Using mice conditionally deficient in both Rap1a and Rap1b and mice null for RIAM, we show that the Rap1/RIAM module is not required for T- or B-cell development but is essential for efficient adhesion to intercellular adhesion molecule (ICAM) 1 and vascular cell adhesion molecule (VCAM) 1 and for proper trafficking of lymphocytes to secondary lymphoid organs. Interestingly, in RIAM-deficient mice, whereas peripheral lymph nodes (pLNs) were depleted of both B and T cells and recirculating B cells were diminished in the bone barrow (BM), the spleen was hypercellular, albeit with a relative deficiency of marginal zone B cells. The abnormality in lymphocyte trafficking was accompanied by defective humoral immunity to T-cell-dependent antigens. Platelet function was intact in RIAM-deficient animals. These in vivo results confirm a role for RIAM in the regulation of some, but not all, leukocyte integrins and suggest that RIAM-regulated integrin activation is required for trafficking of lymphocytes from blood into pLNs and BM, where relatively high shear forces exist in high endothelial venules and sinusoids, respectively.

  18. 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.

  19. Ultrastructural localization of the small GTP-binding protein Rap1 in human platelets and megakaryocytes.

    PubMed

    Berger, G; Quarck, R; Tenza, D; Levy-Toledano, S; de Gunzburg, J; Cramer, E M

    1994-10-01

    Several functions have been proposed for Rap1B in human platelets, including the regulation of phospholipase (PL) C gamma and Ca2+ ATPase. However, its localization is largely unknown. In the present study we have investigated the subcellular distribution of Rap1 by immunocytochemical techniques using affinity purified polyclonal antibodies raised against residues 121-137 common to the 95% homologous Rap1A and Rap1B proteins. By immunofluorescence, a positive labelling was obtained on intact resting platelets and was abolished after adsorption of the antibodies with the control peptide. Immunoelectron microscopy was then used to further define the subcellular localization of Rap1B in platelets and megakaryocytes (MK). In resting cells, immunolabelling for Rap1B was associated with the plasma membrane, mostly at its inner face, and lined the membrane of the open canalicular system (OCS). Some labelling was also found outlining the alpha-granules, identified as such by a double labelling with an anti-GPIIb-IIIa. On thrombasthenic platelets the same localization was observed. When platelets were stimulated by thrombin, immunolabelling for Rap1B was redistributed to the zones of fusion of the granules with the OCS, and to the plasma membrane with a higher concentration on pseudopods. Human MK expressed Rap1 and the staining revealed the association of the protein with the demarcation membranes and alpha-granules. This study presents a first approach to the localization of a small GTP binding-protein Rap1B in whole platelets and MK, and shows its association with both the plasma and OCS membranes, as well as with the alpha-granule membranes.

  20. Downregulation of Rap1GAP in Human Tumor Cells Alters Cell/Matrix and Cell/Cell Adhesion▿ †

    PubMed Central

    Tsygankova, Oxana M.; Ma, Changqing; Tang, Waixing; Korch, Christopher; Feldman, Michael D.; Lv, Yu; Brose, Marcia S.; Meinkoth, Judy L.

    2010-01-01

    Rap1GAP expression is decreased in human tumors. The significance of its downregulation is unknown. We show that Rap1GAP expression is decreased in primary colorectal carcinomas. To elucidate the advantages conferred on tumor cells by loss of Rap1GAP, Rap1GAP expression was silenced in human colon carcinoma cells. Suppressing Rap1GAP induced profound alterations in cell adhesion. Rap1GAP-depleted cells exhibited defects in cell/cell adhesion that included an aberrant distribution of adherens junction proteins. Depletion of Rap1GAP enhanced adhesion and spreading on collagen. Silencing of Rap expression normalized spreading and restored E-cadherin, β-catenin, and p120-catenin to cell/cell contacts, indicating that unrestrained Rap activity underlies the alterations in cell adhesion. The defects in adherens junction protein distribution required integrin signaling as E-cadherin and p120-catenin were restored at cell/cell contacts when cells were plated on poly-l-lysine. Unexpectedly, Src activity was increased in Rap1GAP-depleted cells. Inhibition of Src impaired spreading and restored E-cadherin at cell/cell contacts. These findings provide the first evidence that Rap1GAP contributes to cell/cell adhesion and highlight a role for Rap1GAP in regulating cell/matrix and cell/cell adhesion. The frequent downregulation of Rap1GAP in epithelial tumors where alterations in cell/cell and cell/matrix adhesion are early steps in tumor dissemination supports a role for Rap1GAP depletion in tumor progression. PMID:20439492

  1. 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.

  2. 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.

  3. Phospholipase Cε is a nexus for Rho and Rap-mediated G protein-coupled receptor-induced astrocyte proliferation

    PubMed Central

    Citro, Simona; Malik, Sundeep; Oestreich, Emily A.; Radeff-Huang, Julie; Kelley, Grant G.; Smrcka, Alan V.; Brown, Joan Heller

    2007-01-01

    Phospholipase Cε (PLCε) has been suggested to transduce signals from small GTPases, but its biological function has not yet been clarified. Using astrocytes from PLCε-deficient mice, we demonstrate that endogenous G protein-coupled receptors (GPCRs) for lysophosphatidic acid, sphingosine 1-phosphate, and thrombin regulate phosphoinositide hydrolysis primarily through PLCε. Stimulation by lysophospholipids occurs through Gi, whereas thrombin activates PLC through Rho. Further studies reveal that PLCε is required for thrombin- but not LPA-induced sustained ERK activation and DNA synthesis, providing a novel mechanism for GPCR and Rho signaling to cell proliferation. The requirement for PLCε in this pathway can be explained by its role as a guanine nucleotide exchange factor for Rap1. Thus, PLCε serves to transduce mitogenic signals through a mechanism distinct from its role in generation of PLC-derived second messengers. PMID:17878312

  4. 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.

  5. Mammalian RAP1 controls telomere function and gene expression through binding to telomeric and extra-telomeric sites

    PubMed Central

    Martinez, Paula; Thanasoula, Maria; Carlos, Ana R.; Gómez, Gonzalo; Tejera, Agueda M.; Schoeftner, Stefan; Dominguez, Orlando; Pisano, David G.; Tarsounas, Madalena; Blasco, Maria A.

    2013-01-01

    Shelterin binds and protects mammalian telomeres. Here, we generated cells and mice conditionally deleted for the shelterin component RAP1. We find that Rap1 deficiency is dispensable for telomere capping but leads to increased telomere recombination and fragility. Mice with Rap1 deletion in stratified epithelia are viable but have shorter telomeres and develop skin hyperpigmentation at aduldhood. By performing chromatin immunoprecipitation coupled with ultra-highthroughput sequencing, we find that RAP1 binds to telomeres and to extra-telomeric sites through the (TTAGGG)2 consensus motif. Extra-telomeric RAP1 binding sites are enriched at subtelomeric regions, in agreement with preferential deregulation of subtelomeric genes in Rap1-deficient cells. More than 70% of extra-telomeric RAP1 binding sites are at the vicinity of genes and 31% of the genes deregulated in Rap1-null cells contain RAP1 binding sites, suggesting a role of RAP1 in transcriptional control. These findings place a shelterin component at the interface between telomere function and transcriptional regulation. PMID:20622869

  6. The Rap GTPases regulate the migration, invasiveness and in vivo dissemination of B-cell lymphomas.

    PubMed

    Lin, K B L; Tan, P; Freeman, S A; Lam, M; McNagny, K M; Gold, M R

    2010-01-28

    B-cell lymphomas are common malignancies in which transformed B cells enter the circulation, extravasate into tissues and form tumors in multiple organs. Lymphoma cells are thought to exit the vasculature and enter tissues through the same chemokine- and adhesion molecule-dependent mechanisms as normal B cells. We have previously shown that activation of the Rap GTPases, proteins that control cytoskeletal organization and integrin activation, is critical for chemokine-induced migration and adhesion in B-lymphoma cell lines. Using the A20 murine B-lymphoma cell line as a model, we now show that Rap activation is important for circulating lymphoma cells to enter tissues and form tumors in vivo. In vitro assays showed that Rap activation is required for A20 cells to efficiently adhere to vascular endothelial cells and undergo transendothelial migration. These findings suggest that Rap or its effectors could be novel targets for treating B-cell lymphomas.

  7. Phage Genetic Sites Involved in λ Growth Inhibition by the Escherichia Coli Rap Mutant

    PubMed Central

    Guzman, P.; Guarneros, G.

    1989-01-01

    The rap mutation of Escherichia coli prevents the growth of bacteriophage λ. We have isolated phage mutants that compensate for the host deficiency. The mutations, named bar, were genetically located to three different loci of the λ genome: barI in the attP site, barII in the cIII ea10 region, and barIII within or very near the imm434 region. The level of λ leftward transcription correlates with rap exclusion. Phage λ mutants partially defective in the pL promoter or in pL-transcript antitermination showed a Bar(-) phenotype. Conversely, mutants constitutive for transcription from the pI or pL promoters were excluded more stringently by rap bacteria. We conclude that rap exclusion depends on the magnitude of transcription through the wild type bar loci in the phage genome. PMID:2523838

  8. 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

  9. 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.

  10. Role of nonmuscle myosin IIB and N-RAP in cell spreading and myofibril assembly in primary mouse cardiomyocytes.

    PubMed

    Lu, Shajia; Horowits, Robert

    2008-09-01

    We investigated the role of nonmuscle myosin heavy chain (NMHC) IIB in cultured embryonic mouse cardiomyocytes by specific knockdown using RNA interference. NMHC IIB protein levels decreased 90% compared with mock-transfected cells by 3 days post transfection. NMHC IIB knockdown resulted in a slow decrease in N-RAP protein levels over 6 days with no change in N-RAP transcript levels. N-RAP is a scaffold for alpha-actinin and actin assembly during myofibrillogenesis, and we quantitated myofibril accumulation by morphometric analysis of alpha-actinin organization. Between 3 and 6 days, NMHC IIB knockdown was accompanied by the abolishment of cardiomyocyte spreading. During this period the rate of myofibril accumulation steadily decreased, correlating with the slowly decreasing levels of N-RAP. Between 6 and 8 days NMHC IIB and N-RAP protein levels recovered, and cardiomyocyte spreading and myofibril accumulation resumed. Inhibition of proteasome function using MG132 led to accumulation of excess N-RAP, and the secondary decrease in N-RAP that otherwise accompanied NMHC IIB knockdown was abolished. The results show that NMHC IIB knockdown led to decreased N-RAP levels through proteasome-mediated degradation. Furthermore, these proteins have distinct functional roles, with NMHC IIB playing a role in cardiomyocyte spreading and N-RAP functioning in myofibril assembly.

  11. Modulation of cartilage differentiation by melanoma inhibiting activity/cartilage-derived retinoic acid-sensitive protein (MIA/CD-RAP).

    PubMed

    Schubert, Thomas; Schlegel, Jacqueline; Schmid, Rainer; Opolka, Alfred; Grassel, Susanne; Humphries, Martin; Bosserhoff, Anja-Katrin

    2010-03-31

    Melanoma inhibiting activity/cartilage-derived retinoic acid-sensitive protein (MIA/CD-RAP) is a small soluble protein secreted from malignant melanoma cells and from chondrocytes. Recently, we revealed that MIA/CD-RAP can modulate bone morphogenetic protein (BMP)2-induced osteogenic differentiation into a chondrogenic direction. In the current study we aimed to find the molecular details of this MIA/CD-RAP function. Direct influence of MIA on BMP2 by protein-protein-interaction or modulating SMAD signaling was ruled out experimentally. Instead, we revealed inhibition of ERK signaling by MIA/CD-RAP. This inhibition is regulated via binding of MIA/CD-RAP to integrin alpha5 and abolishing its activity. Active ERK signaling is known to block chondrogenic differentiation and we revealed induction of aggrecan expression in chondrocytes by treatment with MIA/CD-RAP or PD098059, an ERK inhibitor. In in vivo models we could support the role of MIA/CD-RAP in influencing osteogenic differentiation negatively. Further, MIA/CD-RAP-deficient mice revealed an enhanced calcified cartilage layer of the articular cartilage of the knee joint and disordered arrangement of chondrocytes. Taken together, our data indicate that MIA/CD-RAP stabilizes cartilage differentiation and inhibits differentiation into bone potentially by regulating signaling processes during differentiation.

  12. 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.

  13. A Conserved Motif within RAP1 Plays Diversified Roles in Telomere Protection and Regulation in Different Organisms

    PubMed Central

    Chen, Yong; Rai, Rekha; Zhou, Zi-Ren; Kanoh, Junko; Ribeyre, Cyril; Yang, Yuting; Zheng, Hong; Damay, Pascal; Wang, Feng; Tsujii, Hisayo; Hiraoka, Yasushi; Shore, David; Hu, Hong-Yu; Chang, Sandy; Lei, Ming

    2013-01-01

    Repressor activator protein 1 (RAP1) is the most highly conserved telomere protein. It is involved in protecting chromosome ends in fission yeast, promoting gene silencing in Saccharomyces cerevisiae while in Kluyveromyces lactis it is required to repress homology directed recombination (HDR) at telomeres. Since mammalian RAP1 requires TRF2 for stable expression, its role in telomere function has remained obscure. To understand how RAP1 plays such diverse functions at telomeres, we solved the crystal or solution structures of the C-terminal RCT domains of RAP1 from multiple organisms in complex with their respective protein-binding partners. Our comparative structural analysis establishes the RCT domain of RAP1 as an evolutionarily conserved protein-protein interaction module. In mammalian and fission yeast cells, this module interacts with TRF2 and Taz1, respectively, targeting RAP1 to chromosome ends for telomere end protection. While RAP1 repress NHEJ at fission yeast telomeres, at mammalian telomeres it is required to repress HDR. In contrast, S. cerevisiae RAP1 utilizes the RCT domain to recruit Sir3 to telomeres to mediate gene silencing. Together, our results reveal that depending on the organism, the evolutionarily conserved RAP1 RCT motif plays diverse functional roles at telomeres. PMID:21217703

  14. 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.

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

    PubMed Central

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

    2014-01-01

    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 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 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 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. PMID:25236856

  16. Characterization of interactions of adapter protein RAPL/Nore1B with RAP GTPases and their role in T cell migration.

    PubMed

    Miertzschke, Mandy; Stanley, Paula; Bunney, Tom D; Rodrigues-Lima, Fernando; Hogg, Nancy; Katan, Matilda

    2007-10-19

    Using a model of integrin-triggered random migration of T cells, we show that stimulation of LFA-1 integrins leads to the activation of Rap1 and Rap2 small GTPases. We further show that Rap1 and Rap2 have distinct roles in adhesion and random migration of these cells and that an adapter protein from the Ras association domain family (Rassf), RAPL, has a role downstream of Rap2 in addition to its link to Rap1. Further characterization of the RAPL protein and its interactions with small GTPases from the Ras family shows that RAPL forms more stable complexes with Rap2 and classical Ras proteins compared with Rap1. The different interaction pattern of RAPL with Rap1 and Rap2 is not affected by the disruption of the C-terminal SARAH domain that we identified as the alpha-helical region responsible for RAPL dimerization in vitro and in cells. Based on mutagenesis and three-dimensional modeling, we propose that interaction surfaces in RAPL-Rap1 and RAPL-Rap2 complexes are different and that a single residue in the switch I region of Rap proteins (residue 39) contributes considerably to the different kinetics of these protein-protein interactions. Furthermore, the distinct role of Rap2 in migration of T cells is lost when this critical residue is converted to the residue present in Rap1. Together, these observations suggest a wider role for Rassf adapter protein RAPL and Rap GTPases in cell motility and show that subtle differences between highly similar Rap proteins could be reflected in distinct interactions with common effectors and their cellular function.

  17. Overexpression of a novel activator of PAK4, the CDK5 kinase-associated protein CDK5RAP3, promotes hepatocellular carcinoma metastasis.

    PubMed

    Mak, Grace Wing-Yan; Chan, Mandy Man-Lok; Leong, Veronica Yee-Law; Lee, Joyce Man-Fong; Yau, Tai-On; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2011-04-15

    The CDK5 kinase regulatory subunit-associated protein 3 (CDK5RAP3 or C53/LZAP) regulates apoptosis induced by genotoxic stress. Although CDK5RAP3 has been implicated in cancer progression, its exact role in carcinogenesis is not well established. In this article, we report that CDK5RAP3 has an important prometastatic function in hepatocarcinogenesis. An examination of human hepatocellular carcinoma (HCC) samples revealed at least twofold overexpression of CDK5RAP3 transcripts in 58% (39/67) of HCC specimens when compared with corresponding nontumorous livers. CDK5RAP3 overexpression was associated with more aggressive biological behavior. In HCC cell lines, stable overexpression of CDK5RAP3 promoted, and small interfering RNA-mediated knockdown inhibited, tumorigenic activity and metastatic potential. We found that overexpression of CDK5RAP3 and p21-activated protein kinase 4 (PAK4) correlated in human HCCs, and that CDK5RAP3 was a novel binding partner of PAK4, and this binding enhanced PAK4 activity. siRNA-mediated knockdown of PAK4 in CDK5RAP3-expressing HCC cells reversed the enhanced cell invasiveness mediated by CDK5RAP3 overexpression, implying that PAK4 is essential for CDK5RAP3 function. Taken together, our findings reveal that CDK5RAP3 is widely overexpressed in HCC and that overexpression of CDK5RAP3 promotes HCC metastasis through PAK4 activation.

  18. Phosphorylation of Rap1 by cAMP-dependent Protein Kinase (PKA) Creates a Binding Site for KSR to Sustain ERK Activation by cAMP.

    PubMed

    Takahashi, Maho; Li, Yanping; Dillon, Tara J; Stork, Philip J S

    2017-01-27

    Cyclic adenosine monophosphate (cAMP) is an important mediator of hormonal stimulation of cell growth and differentiation through its activation of the extracellular signal-regulated kinase (ERK) cascade. Two small G proteins, Ras and Rap1 have been proposed to mediate this activation. Using HEK293 cells as a model system, we have recently shown that both Ras and Rap1 are required for cAMP signaling to ERKs. However, cAMP-dependent Ras signaling to ERKs is transient and rapidly terminated by PKA phosphorylation of the Raf isoforms C-Raf and B-Raf. In contrast, cAMP-dependent Rap1 signaling to ERKs and Rap1 is potentiated by PKA. We show that this is due to sustained binding of B-Raf to Rap1. One of the targets of PKA is Rap1 itself, directly phosphorylating Rap1a on serine 180 and Rap1b on serine 179. We show that these phosphorylations create potential binding sites for the adaptor protein 14-3-3 that links Rap1 to the scaffold protein KSR. These results suggest that Rap1 activation of ERKs requires PKA phosphorylation and KSR binding. Because KSR and B-Raf exist as heterodimers within the cell, this binding also brings B-Raf to Rap1, allowing Rap1 to couple to ERKs through B-Raf binding to Rap1 independently of its Ras-binding domain.

  19. 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.

  20. 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.

  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. Cdk5rap2 regulates centrosome function and chromosome segregation in neuronal progenitors

    PubMed Central

    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.

    2010-01-01

    Microcephaly affects ∼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 Cdk5rap2an/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. Cdk5rap2an/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

  3. RIAM (Rap1-interacting adaptor molecule) regulates complement-dependent phagocytosis.

    PubMed

    Medraño-Fernandez, Iria; Reyes, Raquel; Olazabal, Isabel; Rodriguez, Elena; Sanchez-Madrid, Francisco; Boussiotis, Vassiliki A; Reche, Pedro A; Cabañas, Carlos; Lafuente, Esther M

    2013-07-01

    Phagocytosis mediated by the complement receptor CR3 (also known as integrin αMß2 or Mac-1) is regulated by the recruitment of talin to the cytoplasmic tail of the ß2 integrin subunit. Talin recruitment to this integrin is dependent on Rap1 activation. However, the mechanism by which Rap1 regulates this event and CR3-dependent phagocytosis remains largely unknown. In the present work, we examined the role of the Rap1 effector RIAM, a talin-binding protein, in the regulation of complement-mediated phagocytosis. Using the human myeloid cell lines HL-60 and THP-1, we determined that knockdown of RIAM impaired αMß2 integrin affinity changes induced by stimuli fMLP and LPS. Phagocytosis of complement-opsonized RBC particles, but not of IgG-opsonized RBC particles, was impaired in RIAM knockdown cells. Rap1 activation via EPAC induced by 8-pCPT-2'-O-Me-cAMP resulted in an increase of complement-mediated phagocytosis that was abrogated by knockdown of RIAM in HL-60 and THP-1 cell lines and in macrophages derived from primary monocytes. Furthermore, recruitment of talin to ß2 integrin during complement-mediated phagocytosis was reduced in RIAM knockdown cells. These results indicate that RIAM is a critical component of the phagocytosis machinery downstream of Rap1 and mediates its function by recruiting talin to the phagocytic complement receptors.

  4. 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.

  5. Celastrol negatively regulates cell invasion and migration ability of human osteosarcoma via downregulation of the PI3K/Akt/NF-κB signaling pathway in vitro

    PubMed Central

    Yu, Xiaolong; Wang, Qiang; Zhou, Xin; Fu, Changlin; Cheng, Ming; Guo, Runsheng; Liu, Hucheng; Zhang, Bin; Dai, Min

    2016-01-01

    Osteosarcoma (OS) is a primary malignant tumor of the bone, with a tendency to metastasize early. Despite the advances in treatment options, more than 30% of patients develop distant metastases, and the prognosis of these patients with metastases is extremely poor. Celastrol has been demonstrated to manifest multiple pharmacological activities, including induction of apoptosis in numerous types of cancer cell lines. Our previous studies have also suggested that Celastrol is capable of inducing apoptosis of human osteosarcoma cells via the mitochondrial-dependent pathway. The purpose of this study was to investigate the effects of Celastrol on the migration and invasion of human osteosarcoma U-2OS cells in vitro. Cell migration and invasion were investigated using wound healing and Boyden chamber Transwell assays. We observed that Celastrol suppressed cell invasion and migration in human osteosarcoma U-2OS cells. Furthermore, protein expression levels of phosphorylated phosphatidylinositol 3-kinase (PI3K), Akt, inhibitor of κB kinase α/β, inhibitor of κB α, nuclear factor-κB (NF-κB subunit p65) and matrix metalloproteinase (MMP)-2 and −9 were measured by western blot analysis. We observed that the PI3K/Akt/NF-κB signaling pathway was inhibited following Celastrol treatment. In addition, the expression levels of MMP-2 and −9 proteins were also reduced significantly following Celastrol treatment. Therefore, we confirmed that Celastrol suppressed osteosarcoma U-2OS cell metastasis via downregulation of the PI3K/Akt/NF-κB signaling pathway in vitro. PMID:27900015

  6. 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

  7. 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…

  8. 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.

  9. Antibodies to the Plasmodium falciparum rhoptry protein RAP-2/RSP-2 in relation to anaemia in Cameroonian children.

    PubMed

    Awah, N; Balogun, H; Achidi, E; Mariuba, L A; Nogueira, P A; Orlandi, P; Troye-Blomberg, M; Gysin, J; Berzins, K

    2011-02-01

    Previous studies have implicated reactive antibodies to the low molecular weight rhoptry-associated proteins (RAP-1, RAP-2/RSP-2 and RAP-3) in erythroid cell destruction during Plasmodium falciparum infection. In this pilot study, the frequency, specificity and functional capacity of naturally acquired anti-RAP-2/RSP-2 antibodies were investigated in the sera of anaemic and nonanaemic malaria-infected Cameroonian children. All sera recognized RAP-2/RSP-2 by FACS, irrespective of the clinical status of the subjects. However, the anaemic children showed higher levels of IgG antibodies than the nonanaemic group, while both groups showed similar levels of IgM antibodies. Only few individuals had detectable levels of RAP-2/RSP-2-specific IgG1 and IgG3 subclass antibodies, while no IgG2 and IgG4 subclass antibodies were detected in these subjects. By ELISA, the anaemic group tended to show higher levels of antibodies to RAP-2/RSP-2 regarding all antibody classes tested, except for IgG4 and IgE. Unexpectedly, sera from the nonanaemic group activated complement to a greater extent than those from the anaemic group. These results need to be confirmed in extended studies but indicate that the effector functions of the RAP-2/RSP-2-reactive antibodies may be more important than their amounts. Such antibodies could play a role in both immunity and pathogenesis during P. falciparum infection.

  10. 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...

  11. 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…

  12. 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

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false May the decision to approve or deny my... ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Remedial Action Plans (RAPs) Getting A Rap Approved § 270.155 May...

  13. 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…

  14. 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

  15. The Relationship between an Oral Rhythmic Style of Communication (Rap Music) and Learning in the Urban Preschool.

    ERIC Educational Resources Information Center

    Hicks, Patricia Thandi

    A study explored the effectiveness of "rap music" as a method of instruction for urban preschool children. It was hypothesized that urban preschool children would learn more new content (10 unfamiliar names of body parts) in a classroom environment through the use of rap music for instruction, than would children who received instruction…

  16. 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]…

  17. Explicit Rap Music Lyrics and Attitudes toward Rape: The Perceived Effects on African American College Students' Attitudes.

    ERIC Educational Resources Information Center

    Wade, Bruce H.; Thomas-Gunnar, Cynthia A.

    1993-01-01

    Examines the effects of rap music on the attitudes and behaviors of students in historically black colleges. Interviews with 38 females indicate that they find explicit lyrics inappropriate and harmful to society, but they feel that rap music accurately represents some of the realities of gender relations between black males and females. (SLD)

  18. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei.

    PubMed

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

    2009-04-03

    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, 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.

  19. 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.

  20. The relationship between heavy metal and rap music and adolescent turmoil: real or artifact?

    PubMed

    Took, K J; Weiss, D S

    1994-01-01

    Adolescents and their parents were surveyed to investigate the association between heavy metal and rap music and adolescent psychosocial turmoil. Subjects were asked about current and past psychosocial functioning, as well as their music preferences. Adolescents who preferred heavy metal and rap music were compared with those who preferred other types of music. Results indicated that adolescents who preferred heavy metal and rap had a higher incidence of below-average school grades, school behavior problems, sexual activity, drug and alcohol use, and arrests. However, when gender was controlled, only below-average current and elementary school grades and a history of counseling in elementary school for school problems remained significant. Implications of these findings are discussed.

  1. 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... management activities under a RAP at a location removed from the area where the remediation wastes originated? (a) You may request a RAP for remediation waste management activities at a location removed from...

  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... management activities under a RAP at a location removed from the area where the remediation wastes originated? (a) You may request a RAP for remediation waste management activities at a location removed from...

  3. Development of RAP Tag, a Novel Tagging System for Protein Detection and Purification.

    PubMed

    Fujii, Yuki; Kaneko, Mika K; Ogasawara, Satoshi; Yamada, Shinji; Yanaka, Miyuki; Nakamura, Takuro; Saidoh, Noriko; Yoshida, Kanae; Honma, Ryusuke; Kato, Yukinari

    2017-03-24

    Affinity tag systems, possessing high affinity and specificity, are useful for protein detection and purification. The most suitable tag for a particular purpose should be selected from many available affinity tag systems. In this study, we developed a novel affinity tag called the "RAP tag" system, which comprises a mouse antirat podoplanin monoclonal antibody (clone PMab-2) and the RAP tag (DMVNPGLEDRIE). This system is useful not only for protein detection in Western blotting, flow cytometry, and sandwich enzyme-linked immunosorbent assay, but also for protein purification.

  4. 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

  5. DIXDC1 increases the invasion and migration ability of non-small-cell lung cancer cells via the PI3K-AKT/AP-1 pathway.

    PubMed

    Xu, Zhonghai; Liu, Di; Fan, Chuifeng; Luan, Lan; Zhang, Xiupeng; Wang, Enhua

    2014-11-01

    DIX domain containing 1 (DIXDC1), is a human homolog of Ccd1, a recently identified DIX domain containing protein in zebrafish. DIXDC1 protein was detected in human colorectal adenocarcinoma tissues and was found to be correlated with a high cell proliferation index. We demonstrated DIXDC1 overexpression in 55% (92/167) of non-small cell lung cancer (NSCLC) cases, compared to adjacent noncancerous lung tissues (P < 0.01). Overexpression of DIXDC1 was associated with lymph node metastasis and more advanced TNM stage (P < 0.001 and P = 0.001, respectively). Kaplan-Meier survival curves and log-rank testing indicated that overexpression of DIXDC1 correlated with worse overall survival in NSCLC (P = 0.031). DIXDC1 was more abundant in seven NSCLC lines than the bronchial cell line HBE, and modulation of its expression regulated AP-1 activity; MMP2, MMP7, and MMP9 protein and mRNA; and invasion ability. Metalloproteinase induction was reversed by PI3K/AKT and AP-1 inhibition. These results suggest DIXDC1 is associated with stage and prognosis in NSCLC, and may promote invasion and migration through PI3K-AKT/AP-1-dependent activation of metalloproteinases.

  6. Receptor-associated protein (RAP) has two high-affinity binding sites for the low-density lipoprotein receptor-related protein (LRP): consequences for the chaperone functions of RAP.

    PubMed

    Jensen, Jan K; Dolmer, Klavs; Schar, Christine; Gettins, Peter G W

    2009-06-26

    RAP (receptor-associated protein) is a three domain 38 kDa ER (endoplasmic reticulum)-resident protein that is a chaperone for the LRP (low-density lipoprotein receptor-related protein). Whereas RAP is known to compete for binding of all known LRP ligands, neither the location, the number of binding sites on LRP, nor the domains of RAP involved in binding is known with certainty. We have systematically examined the binding of each of the three RAP domains (D1, D2 and D3) to tandem and triple CRs (complement-like repeats) that span the principal ligand-binding region, cluster II, of LRP. We found that D3 binds with low nanomolar affinity to all (CR)2 species examined. Addition of a third CR domain increases the affinity for D3 slightly. A pH change from 7.4 to 5.5 gave only a 6-fold increase in Kd for D3 at 37 degrees C, whereas temperature change from 22 degrees C to 37 degrees C has a similar small effect on affinity, raising questions about the recently proposed D3-destabilization mechanism of RAP release from LRP. Surprisingly, and in contrast to literature suggestions, D1 and D2 also bind to most (CR)2 and (CR)3 constructs with nanomolar affinity. Although this suggested that there might be three high-affinity binding sites in RAP for LRP, studies with intact RAP showed that only two binding sites are available in the intact chaperone. These findings suggest a new model for RAP to function as a folding chaperone and also for the involvement of YWTD domains in RAP release from LRP in the Golgi.

  7. Clinical analysis of the rap stress stimulator applied for crus fracture after skeletal external fixation

    PubMed Central

    Zhuang, Ping; Hong, Jiayuan; Chen, Wei; Wu, Jin

    2015-01-01

    Introduction Open crus fracture is still difficult in clinical treatment because of the delayed fracture union and high rate of nonunion after the operation. A consensus has been reached that mechanical stress can promote fracture healing. We independently developed a stress stimulator, which can provide longitudinal pressure for the fixed fracture end of the lower legs to promote fracture healing. The purpose of this study is to explore the advantages and clinical effect of the rap stress stimulator applied for open crus fracture after skeletal external fixation. Material and methods One hundred and sixty-five patients (183 limbs) who suffered from open tibia and fibula fracture received skeletal external fixation, of which 108 limbs were treated with the rap stress stimulator after external fixation and 75 limbs were treated with regular functional exercises of muscle contraction and joint activity only. Then the fracture healing time and rate of nonunion were compared between the two groups. Results The mean fracture healing time and rate of nonunion in the group treated with the rap stress stimulator were 138.27 ±4.45 days and 3.70% respectively, compared to 153.43 ±4.89 days and 10.67% in the group treated without the stimulator. Conclusions The rap stress stimulator significantly shortened the fracture healing time and reduced the rate of nonunion for treating open tibia and fibula fractures. PMID:26170856

  8. ABNORMAL GASTRIC AND COLONIC PERMEABILITY IN CHILDREN WITH RECURRENT ABDOMINAL PAIN (RAP)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent histologic studies have suggested evidence of low grade inflammation in many patients with irritable bowel syndrome (IBS). Additionally, small intestinal permeability recently has been reported to be abnormal in some adults with IBS. Whether the same is true for children with RAP, a condition...

  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. 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…

  11. 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

  12. 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.

  13. 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…

  14. 40 CFR 270.110 - What must I include in my application for a RAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...? You must include the following information in your application for a RAP: (a) The name, address, and EPA identification number of the remediation waste management site; (b) The name, address, and... States Geological Survey (USGS) or county map showing the location of the remediation waste...

  15. 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…

  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. 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…

  18. "Fight the Power" : Rap Music Pounds Out a New Anthem for Many Black Students.

    ERIC Educational Resources Information Center

    Collison, Michele N-K

    1990-01-01

    Black college students are increasingly embracing the message of rap music and challenging the status quo. While not all students share the view, students advocating self-determination and emphasizing their African roots have become a vocal majority among Blacks on many campuses. (MSE)

  19. The Relationship between Types of Rap Music and Memory in African American Children.

    ERIC Educational Resources Information Center

    Hall, Pamela D.

    1998-01-01

    Studied the types of rap music that are most understandable and the age groups most likely to be affected with 30 children, half ages 7 to 9 and the others ages 10 to 12 years. Younger children could not always describe what the music was about, but they did have a general understanding. (SLD)

  20. 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…

  1. 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…

  2. A Musical Color Line: The Problem of Race in White Rap Rhetoric.

    ERIC Educational Resources Information Center

    Riddles, Allison

    An instructor of freshman composition at Ohio University always teaches a section on music in her courses because freshmen jump at the chance to discuss a part of their youth culture that they readily identify with. The problem, however, has been how to incorporate rap music successfully into these discussions with a classroom full of white…

  3. 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,…

  4. 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…

  5. 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…

  6. The Influence of Misogynous Rap Music on Sexual Aggression against Women.

    ERIC Educational Resources Information Center

    Barongan, Christy; Hall, Gordon C. Nagayama

    1995-01-01

    Results from 54 college men who heard misogynous or neutral rap music and then selected neutral, sexual-violent, or assaultive film vignettes to show a female companion suggest that misogynous music facilitates sexually aggressive behavior and support a relationship between cognitive distortions concerning women and sexual aggression. (SLD)

  7. 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)

  8. 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

  9. The role of RAP1 in the regulation of the MAT alpha locus.

    PubMed Central

    Giesman, D; Best, L; Tatchell, K

    1991-01-01

    The RAP1 gene of Saccharomyces cerevisiae encodes an abundant DNA-binding protein, also known as GRF1, TBA, or TUF, that binds to many sites in the yeast genome in vitro. These sites define a consensus sequence, [sequence: see text], and deletion analyses of genes that contain this sequence have implicated the involvement of RAP1 in numerous cellular processes, including gene activation and repression. The MAT alpha locus, required for determination of the alpha cell type in yeast cells, contains a RAP1 binding site; this site coincides with the MAT alpha upstream activating sequence (UAS) and is necessary for expression of the two genes encoded by the MAT alpha locus, MAT alpha 1 and MAT alpha 2. We show that the MAT alpha UAS is sufficient to activate transcription from a promoterless gene fusion of the yeast CYC1 upstream region and the lacZ gene. Constructs containing only the MAT alpha UAS generated elevated levels of beta-galactosidase activity which were indistinguishable from those of constructs containing the entire MAT alpha intergenic region. Further, the MAT alpha UAS has an intrinsic polarity of transcriptional activation; transcription of CYC1-lacZ was six- to sevenfold higher when the UAS was oriented in the direction normally associated with MAT alpha 2 transcription. Point mutations in the MAT alpha UAS that reduce MAT alpha expression three- to fivefold resulted in a bi-mating phenotype, while a mutation that reduced MAT alpha expression still further resulted in an a-mating phenotype. We isolated plasmids from a high-copy-number yeast library that suppressed the bi-mating defect of point mutations in the MAT alpha UAS, and the most effective dosage suppressor contained the gene encoding RAP1. A temperature-sensitive rap1 mutant bi-mates at the semipermissive temperature. Double mutants at rap1 and mat alpha mate exclusively as a cells, at all temperatures, and do not express detectable levels of MAT alpha RNA. These data provide evidence that the

  10. cAMP-induced Epac-Rap activation inhibits epithelial cell migration by modulating focal adhesion and leading edge dynamics.

    PubMed

    Lyle, Karen S; Raaijmakers, Judith H; Bruinsma, Wytse; Bos, Johannes L; de Rooij, Johan

    2008-06-01

    Epithelial cell migration is a complex process crucial for embryonic development, wound healing and tumor metastasis. It depends on alterations in cell-cell adhesion and integrin-extracellular matrix interactions and on actomyosin-driven, polarized leading edge protrusion. The small GTPase Rap is a known regulator of integrins and cadherins that has also been implicated in the regulation of actin and myosin, but a direct role in cell migration has not been investigated. Here, we report that activation of endogenous Rap by cAMP results in an inhibition of HGF- and TGFbeta-induced epithelial cell migration in several model systems, irrespective of the presence of E-cadherin adhesion. We show that Rap activation slows the dynamics of focal adhesions and inhibits polarized membrane protrusion. Importantly, forced integrin activation by antibodies does not mimic these effects of Rap on cell motility, even though it does mimic Rap effects in short-term cell adhesion assays. From these results, we conclude that Rap inhibits epithelial cell migration, by modulating focal adhesion dynamics and leading edge activity. This extends beyond the effect of integrin affinity modulation and argues for an additional function of Rap in controlling the migration machinery of epithelial cells.

  11. The small GTPase Rap1b negatively regulates neutrophil chemotaxis and transcellular diapedesis by inhibiting Akt activation.

    PubMed

    Kumar, Sachin; Xu, Juying; Kumar, Rupali Sani; Lakshmikanthan, Sribalaji; Kapur, Reuben; Kofron, Matthew; Chrzanowska-Wodnicka, Magdalena; Filippi, Marie-Dominique

    2014-08-25

    Neutrophils are the first line of cellular defense in response to infections and inflammatory injuries. However, neutrophil activation and accumulation into tissues trigger tissue damage due to release of a plethora of toxic oxidants and proteases, a cause of acute lung injury (ALI). Despite its clinical importance, the molecular regulation of neutrophil migration is poorly understood. The small GTPase Rap1b is generally viewed as a positive regulator of immune cell functions by controlling bidirectional integrin signaling. However, we found that Rap1b-deficient mice exhibited enhanced neutrophil recruitment to inflamed lungs and enhanced susceptibility to endotoxin shock. Unexpectedly, Rap1b deficiency promoted the transcellular route of diapedesis through endothelial cell. Increased transcellular migration of Rap1b-deficient neutrophils in vitro was selectively mediated by enhanced PI3K-Akt activation and invadopodia-like protrusions. Akt inhibition in vivo suppressed excessive Rap1b-deficient neutrophil migration and associated endotoxin shock. The inhibitory action of Rap1b on PI3K signaling may be mediated by activation of phosphatase SHP-1. Thus, this study reveals an unexpected role for Rap1b as a key suppressor of neutrophil migration and lung inflammation.

  12. Rap1 in Candida albicans: an Unusual Structural Organization and a Critical Function in Suppressing Telomere Recombination▿

    PubMed Central

    Yu, Eun Young; Yen, Wei-Feng; Steinberg-Neifach, Olga; Lue, Neal F.

    2010-01-01

    Rap1 (repressor activator protein 1) is a conserved multifunctional protein initially identified as a transcriptional regulator of ribosomal protein genes in Saccharomyces cerevisiae but subsequently shown to play diverse functions at multiple chromosomal loci, including telomeres. The function of Rap1 appears to be evolutionarily plastic, especially in the budding yeast lineages. We report here our biochemical and molecular genetic characterizations of Candida albicans Rap1, which exhibits an unusual, miniaturized domain organization in comparison to the S. cerevisiae homologue. We show that in contrast to S. cerevisiae, C. albicans RAP1 is not essential for cell viability but is critical for maintaining normal telomere length and structure. The rap1 null mutant exhibits drastic telomere-length dysregulation and accumulates high levels of telomere circles, which can be largely attributed to aberrant recombination activities at telomeres. Analysis of combination mutants indicates that Rap1 and other telomere proteins mediate overlapping but nonredundant roles in telomere protection. Consistent with the telomere phenotypes of the mutant, C. albicans Rap1 is localized to telomeres in vivo and recognizes the unusual telomere repeat unit with high affinity and sequence specificity in vitro. The DNA-binding Myb domain of C. albicans Rap1 is sufficient to suppress most of the telomere aberrations observed in the null mutant. Notably, we were unable to detect specific binding of C. albicans Rap1 to gene promoters in vivo or in vitro, suggesting that its functions are more circumscribed in this organism. Our findings provide insights on the evolution and mechanistic plasticity of a widely conserved and functionally critical telomere component. PMID:20008550

  13. Comparative Evaluation of the BD Phoenix Yeast ID Panel and Remel RapID Yeast Plus System for Yeast Identification.

    PubMed

    Grant, Michelle L; Parajuli, Shobha; Deleon-Gonsalves, Raquel; Potula, Raghava; Truant, Allan L

    2016-01-01

    Becton Dickinson Phoenix Yeast ID Panel was compared to the Remel RapID Yeast Plus System using 150 recent clinical yeast isolates and the API 20C AUX system to resolve discrepant results. The concordance rate between the Yeast ID Panel and the RapID Yeast Plus System (without arbitration) was 93.3% with 97.3% (146/150) and 95.3% (143/150) of the isolates correctly identified by the Becton Dickinson Phoenix and the Remel RapID, respectively, with arbitration.

  14. Comparative Evaluation of the BD Phoenix Yeast ID Panel and Remel RapID Yeast Plus System for Yeast Identification

    PubMed Central

    Grant, Michelle L.; Parajuli, Shobha; Deleon-Gonsalves, Raquel; Potula, Raghava; Truant, Allan L.

    2016-01-01

    Becton Dickinson Phoenix Yeast ID Panel was compared to the Remel RapID Yeast Plus System using 150 recent clinical yeast isolates and the API 20C AUX system to resolve discrepant results. The concordance rate between the Yeast ID Panel and the RapID Yeast Plus System (without arbitration) was 93.3% with 97.3% (146/150) and 95.3% (143/150) of the isolates correctly identified by the Becton Dickinson Phoenix and the Remel RapID, respectively, with arbitration. PMID:27366167

  15. Musical ability.

    PubMed

    Sloboda, J

    1993-01-01

    Musical ability is the ability to 'make sense' of music, and develops in most people over the first decade of life through normal enculturation. Whether this ability is developed to a high level usually depends on the decision to start learning a musical instrument, which forces high levels of focused cognitive engagement (practice) with musical materials. Performance ability has both technical and expressive aspects. These aspects are not always developed equally well. Factors contributing to the development of a well-balanced musical performer include (a) lengthy periods of engagement with music through practice and exploration, (b) high levels of material and emotional support from parents and other adults, (c) relationships with early teachers characterized by warmth and mutual liking, and (d) early experiences with music that promote, rather than inhibit, intense sensuous/affective experiences. It is argued that much formal education inhibits the development of musical ability through over-emphasis on assessment, creating performance anxiety, coupled with class and sex stereotyping of approved musical activities. Early free exploration of a medium is a necessity for the development of high levels of musicality.

  16. Interspecies Complementation of the LuxR Family Pathway-Specific Regulator Involved in Macrolide Biosynthesis.

    PubMed

    Mo, SangJoon; Yoon, Yeo Joon

    2016-01-01

    PikD is a widely known pathway-specific regulator for controlling pikromycin production in Streptomyces venezuelae ATCC 15439, which is a representative of the large ATP-binding regulator of the LuxR family (LAL) in Streptomyces sp. RapH and FkbN also belong to the LAL family of transcriptional regulators, which show greatest homology with the ATP-binding motif and helix-turn-helix DNA-binding motif of PikD. Overexpression of pikD and heterologous expression of rapH and fkbN led to enhanced production of pikromycin by approximately 1.8-, 1.6-, and 1.6-fold in S. venezuelae, respectively. Cross-complementation of rapH and fkbN in the pikD deletion mutant (ΔpikD) restored pikromycin and derived macrolactone production. Overall, these results show that heterologous expression of rapH and fkbN leads to the overproduction of pikromycin and its congeners from the pikromycin biosynthetic pathway in S. venezuelae, and they have the same functionality as the pathwayspecific transcriptional activator for the pikromycin biosynthetic pathway in the ΔpikD strain. These results also show extensive "cross-communication" between pathway-specific regulators of streptomycetes and suggest revision of the current paradigm for pathwayspecific versus global regulation of secondary metabolism in Streptomyces species.

  17. 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.

  18. 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.

  19. 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.

  20. Feeling the beat: the meaning of rap music for ethnically diverse Midwestern college students--a phenomenological study.

    PubMed

    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 negative aspects of the music; there has been a dearth of in-depth qualitative studies that explore how rap impacts the listener. Our phenomenological study explores that impact on ethnically diverse college students. Results indicate a profound psychological and educational effect and the discussion goes on to highlight the potential and innovative ways rap music can be utilized with adolescents in fields such as education, risk reduction programs, and counseling psychology.

  1. 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.

  2. Relationship between Rap1 protein phosphorylation and regulation of Ca2+ transport in platelets: a new approach.

    PubMed Central

    Magnier, C; Corvazier, E; Aumont, M C; Le Jemtel, T H; Enouf, J

    1995-01-01

    Although the interrelationship between the two messengers Ca2+ and cyclic AMP in platelet function is well documented, its mechanism of action still remains to be established. We investigated here the question of the regulation of platelet Ca(2+)-ATPases by cyclic AMP through the phosphorylation of the Rap1 protein using a pathological model. We first found experimental conditions where Ca(2+)-transport by platelet membrane vesicles appeared to be dependent on the phosphorylation of the Rap1 protein. Then, we studied platelets of patients with congestive heart failure for their expression of the potential 97 kDa Ca(2+)-ATPase target of regulation through the Rap1 protein as well as the phosphorylation of the Rap1 protein using the catalytic subunit of the cyclic AMP-dependent protein kinase (C. Sub.). In the first patients studied, we found no significant modification in the expression of the 97 kDa Ca(2+)-ATPase by Western blotting using the PL/IM 430 monoclonal antibody which specifically recognized this isoform. In contrast, the Rap1 protein was differentially phosphorylated when using 15 micrograms/ml of the C. Sub. These results allowed us to use these pathological platelets to study the relationship between the expression of Rap1 protein and the regulation of Ca2+ transport by selecting a patient with severe heart failure. We could show a decrease in the expression as well as in the phosphorylation of Rap1 protein and demonstrate a lower effect of C. Sub. on Ca2+ transport. Finally, by studying a further series of patients, we could confirm that the decrease in Rap1 protein expression in heart failure, whatever its extent, was variable, and could strictly correlate the expression of Rap1 protein with the stimulatory effect of C. Sub. on Ca2+ transport. Besides the evidence for regulation of the expression of the Rap1 protein in platelets from patients with heart failure, these findings constitute a new approach in favour of the regulation of platelet Ca2

  3. A genetic modifier screen identifies multiple genes that interact with Drosophila Rap/Fzr and suggests novel cellular roles.

    PubMed

    Kaplow, Margarita E; Mannava, Laura J; Pimentel, Angel C; Fermin, Hector A; Hyatt, Vanetta J; Lee, John J; Venkatesh, Tadmiri R

    2007-01-01

    In the developing Drosophila eye, Rap/Fzr plays a critical role in neural patterning by regulating the timely exit of precursor cells. Rap/Fzr (Retina aberrant in pattern/Fizzy related) is an activator of the E3 Ubiquitin ligase, the APC (Anaphase Promoting Complex-cyclosome) that facilitates the stage specific proteolytic destruction of mitotic regulators, such as cyclins and cyclin-dependent kinases. To identify novel functional roles of Rap/Fzr, we conducted an F(1) genetic modifier screen to identify genes which interact with the partial-loss-function mutations in rap/fzr. We screened 2741 single P-element, lethal insertion lines and piggyBac lines on the second and third chromosome for dominant enhancers and suppressors of the rough eye phenotype of rap/fzr. From this screen, we have identified 40 genes that exhibit dosage-sensitive interactions with rap/fzr; of these, 31 have previously characterized cellular functions. Seven of the modifiers identified in this study are regulators of cell cycle progression with previously known interactions with rap/fzr. Among the remaining modifiers, 27 encode proteins involved in other cellular functions not directly related to cell-cycle progression. The newly identified variants fall into at least three groups based on their previously known cellular functions: transcriptional regulation, regulated proteolysis, and signal transduction. These results suggest that, in addition to cell cycle regulation, rap/fzr regulates ubiquitin-ligase-mediated protein degradation in the developing nervous system as well as in other tissues.

  4. Platelet sarco/endoplasmic reticulum Ca2+ATPase isoform 3b and Rap 1b: interrelation and regulation in physiopathology.

    PubMed Central

    Lacabaratz-Porret, C; Corvazier, E; Kovàcs, T; Bobe, R; Bredoux, R; Launay, S; Papp, B; Enouf, J

    1998-01-01

    Platelet Ca2+ signalling involves intracellular Ca2+ pools, whose content is controlled by sarco/endoplasmic reticulum Ca2+ATPases (SERCAs). Among these, a key role is played by the inositol trisphosphate-sensitive Ca2+ pool, associated with the SERCA 3b isoform. We have investigated the control of this Ca2+ pool through the cAMP-dependent phosphorylation of the GTP-binding protein, Rap (Ras-proximate) 1b. We first looked for this Ca2+ pool target of regulation by studying the expression of the different SERCA and Rap 1 proteins in human platelets and various cell lines, by Western blotting and reverse transcription-PCR. Since co-expression of Rap 1b and SERCA 3b was obtained, we looked for their protein-protein interaction as a function of the cAMP-dependent phosphorylation of Rap 1b. Co-immunoprecipitations of SERCA 3b and Rap 1b proteins were found in the absence of phosphorylation, induced by the catalytic subunit of the cAMP-dependent protein kinase (csPKA). In contrast, upon pre-treatment of platelet membranes with csPKA, the SERCA 3b dissociated from the Rap 1b protein, in agreement with a role of its phosphorylated state in their interaction. Finally, we looked for adaptation of this complex in a platelet pathological model of hypertension. We investigated the expression of both proteins, as well as the cAMP-dependent phosphorylation of Rap 1b and SERCA 3b activity in platelets from control normotensive Wistar-Kyoto rats and from spontaneously hypertensive rats (SHRs). A decrease in SERCA 3b activity was associated with a decrease in Rap 1b endogenous phosphorylation in SHR platelets, consistent with a functional role in the regulation of the SERCA 3b-associated Ca2+ pool. PMID:9576865

  5. 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.

  6. "Rap Universal": Using Multimodal Media Production to Develop ICT Literacies

    ERIC Educational Resources Information Center

    Turner, K. C. Nat

    2011-01-01

    Through a multimodal media production literacy intervention in an extended-day program, culturally and linguistically diverse youth developed valuable information and communication technology literacies, including: (1) Specific how-to skills useful in future academic, professional, social, and civic contexts; (2) Abilities to critically interpret…

  7. 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

  8. 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.

  9. [Effects of lead and selenium on telomere binding protein Rap1p, telomerase and telomeric DNA in Saccharomyces cerevisiae].

    PubMed

    Cui, Qing-Hua; Tang, Chia-Chun; Huang, You-Guo

    2002-03-01

    The effects on S.cerevisiae telomere binding protein Rap1p, telomerase and telomeric DNA by the lead (Pb), the selenium (Se) and Pb + Se were tested respectively in this study. Compared with the control S.cerevisiae after 100 gene rations, the mean telomere length shortened, Rap1p concentration was significantly lower and the secondary structure of Rap1p was disturbed, the telomerase activity was reduced in Pb treated cells. In Se treated cells, telomere length was significantly longer, and telomerase activity expressed higher. The concentration and secondary structure of Rap1p were similar to that of the control. Further more, the viability of Pb treated cells were significantly reduced while cells undergone other three treatments were similar and normal. These results suggest that Pb could damage Rap1p, reduce telomerase activity, resulting in the telomer length shortening and cell death. On the other hand, Se could protect and repair the damage in Rap1p and telomere caused by Pb to some extent.

  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. 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.

  12. Rap1: a turnabout for the crosstalk between cadherins and integrins.

    PubMed

    Retta, Saverio Francesco; Balzac, Fiorella; Avolio, Maria

    2006-04-01

    The coordinate modulation of the cellular functions of cadherins and integrins plays an essential role in fundamental physiological and pathological processes, including morphogenesis, tissue differentiation and renewal, wound healing, immune surveillance, inflammatory response, tumour progression, and metastasis. However, the molecular mechanisms underlying the fine-balanced relationship between cadherin and integrin functions are still elusive. This review focuses on recent findings on the involvement of the small GTPase Rap1 in the regulation of cadherin- and integrin-dependent cell adhesion and signal transduction. In particular, it highlights some of the novel results recently obtained that raise the possibility of a pivotal role for Rap1 in the functional crosstalk between cadherins and integrins, suggesting interesting new regulatory mechanisms.

  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. 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.

  15. 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.

  16. The Recruit Assessment Program (RAP) Experience With Adverse Childhood Experiences (ACE) Questions

    DTIC Science & Technology

    2006-02-22

    recruits in San Diego. It differs from previous versions in that it asks additional education questions, such as history of homeschooling , school...Albuquerque, New Mexico . 17. Landis JR, Koch GG. The measurement of observer agreement for categorical data. Biometrics. 1977;33(1):159–174. 18...Protection Conference, August 11–17, 2003, Albuquerque, New Mexico . 14 15 Table 4. Top 10 RAP Questions Fort Jackson, SC, Participants “Skipped or

  17. 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.

  18. 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.

  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. Dipole localization using beamforming and RAP-MUSIC on simulated intracerebral recordings.

    PubMed

    Chang, N; Gotman, J; Gulrajani, R

    2004-01-01

    Interpreting intracerebral recordings in the search of an epileptic focus can be difficult because the amplitude of the potentials are misleading. Small generators located near the electrode site generate large potentials, which could swamp the signal of a nearby epileptic focus. In order to address this problem, two inverse problem algorithms, beamforming and recursively applied and projected multiple signal classification (RAP-MUSIC), were used with simulated intracerebral potentials to calculate equivalent dipole positions. Three dipoles were positioned in an infinite plane medium near three intracerebral electrodes. The potentials generated by the dipoles were simulated and contaminated with white noise. Initial localization simulations showed that both methods detected the sources accurately with RAP-MUSIC reporting lower orientation errors. A spatial resolution analysis for both methods was undertaken in which two dipoles were placed on a plane with the same orientation and overlapping time-courses. Beamforming was able to adequately distinguish the sources for separation distances of 1.2 cm, whereas RAP-MUSIC managed to separate the sources for dipoles as close as 0.4-0.6 cm.

  1. 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.

  2. 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.

  3. miR-337-3p and Its Targets STAT3 and RAP1A Modulate Taxane Sensitivity in Non-Small Cell Lung Cancers

    PubMed Central

    Du, Liqin; Subauste, Maria C.; DeSevo, Christopher; Zhao, Zhenze; Baker, Michael; Borkowski, Robert; Schageman, Jeoffrey J.; Greer, Rachel; Yang, Chin-Rang; Suraokar, Milind; Wistuba, Ignacio I.; Gazdar, Adi F.; Minna, John D.; Pertsemlidis, Alexander

    2012-01-01

    NSCLC (non-small cell lung cancer) often exhibits resistance to paclitaxel treatment. Identifying the elements regulating paclitaxel response will advance efforts to overcome such resistance in NSCLC therapy. Using in vitro approaches, we demonstrated that over-expression of the microRNA miR-337-3p sensitizes NCI-H1155 cells to paclitaxel, and that miR-337-3p mimic has a general effect on paclitaxel response in NSCLC cell lines, which may provide a novel adjuvant strategy to paclitaxel in the treatment of lung cancer. By combining in vitro and in silico approaches, we identified STAT3 and RAP1A as direct targets that mediate the effect of miR-337-3p on paclitaxel sensitivity. Further investigation showed that miR-337-3p mimic also sensitizes cells to docetaxel, another member of the taxane family, and that STAT3 levels are significantly correlated with taxane resistance in lung cancer cell lines, suggesting that endogenous STAT3 expression is a determinant of intrinsic taxane resistance in lung cancer. The identification of a miR-337-3p as a modulator of cellular response to taxanes, and STAT3 and RAP1A as regulatory targets which mediate that response, defines a novel regulatory pathway modulating paclitaxel sensitivity in lung cancer cells, which may provide novel adjuvant strategies along with paclitaxel in the treatment of lung cancer and may also provide biomarkers for predicting paclitaxel response in NSCLC. PMID:22723956

  4. 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

  5. Molecular characterization and chromosomal assignment of equine cartilage derived retinoic acid sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA).

    PubMed

    Berg, Lise C; Mata, Xavier; Thomsen, Preben D

    2008-01-15

    Cartilage-derived retinoic acid sensitive protein (CD-RAP) also known as melanoma inhibitory activity (MIA) has already been established as a marker for chondrocyte differentiation and a number of cancerous conditions in humans. Studies have also shown that CD-RAP/MIA is a potential marker of joint disease. The objective of this study was to characterize the equine CD-RAP/MIA gene and thus make it available as a marker in cartilage research and clinical studies. Gene analysis revealed that the equine gene (GenBank accession no. EF679787) consists of four exons and three introns, and the homology to the human gene is 90% for the translated region. The upstream sequence includes regulatory elements and putative transcription factor binding sites previously described in the human and murine promoter regions. The deduced amino acid sequence consists of 130 aa including a signal peptide of 23 aa, and has a 91% identity to the human protein. Using radiation hybrid mapping, the CD-RAP/MIA gene was localized to the p arm of equine chromosome 10 (ECA10p), which is in accordance with prediction based on the current human-equine comparative map. Gene expression studies showed expression of CD-RAP/MIA mRNA in articular cartilage and chondrocytes from horses with no signs of joint disease. The expression decreased as the cells dedifferentiated in monolayer culture. We also identified an equine CD-RAP/MIA splice variant similar to that reported in humans. The CD-RAP/MIA protein was detected in equine synovial fluid, serum and culture medium from chondrocyte cultures. In conclusion, CD-RAP/MIA is expressed in equine cartilage and chondrocytes, and the protein can be detected in equine serum, synovial fluid and in culture medium from chondrocyte cultures. The equine gene and resulting protein share great homology with the human gene, making future studies on CD-RAP/MIAs potential as a marker in joint disease possible using the equine joint as a model.

  6. Evidence for negative selection on the gene encoding rhoptry-associated protein 1 (RAP-1) in Plasmodium spp.

    PubMed

    Pacheco, M Andreína; Ryan, Elizabeth M; Poe, Amanda C; Basco, Leonardo; Udhayakumar, Venkatachalam; Collins, Williams E; Escalante, Ananias A

    2010-07-01

    Assessing how natural selection, negative or positive, operates on genes with low polymorphism is challenging. We investigated the genetic diversity of orthologous genes encoding the rhoptry-associated protein 1 (RAP-1), a low polymorphic protein of malarial parasites that is involved in erythrocyte invasion. We applied evolutionary genetic methods to study the polymorphism in RAP-1 from Plasmodium falciparum (n=32) and Plasmodium vivax (n=6), the two parasites responsible for most human malaria morbidity and mortality, as well as RAP-1 orthologous in closely related malarial species found in non-human primates (NHPs). Overall, genes encoding RAP-1 are highly conserved in all Plasmodium spp. included in this investigation. We found no evidence for natural selection, positive or negative, acting on the gene encoding RAP-1 in P. falciparum or P. vivax. However, we found evidence that the orthologous genes in non-human primate parasites (Plasmodium cynomolgi, Plasmodium inui, and Plasmodium knowlesi) are under purifying (negative) selection. We discuss the importance of considering negative selection while studying genes encoding proteins with low polymorphism and how selective pressures may differ among orthologous genes in closely related malarial parasites species.

  7. 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.

  8. Comparative performance of the RapID Yeast Plus System and the API 20C AUX Clinical Yeast System.

    PubMed

    Smith, M B; Dunklee, D; Vu, H; Woods, G L

    1999-08-01

    The performance of the RapID Yeast Plus System (Innovative Diagnostic Systems, Norcross, Ga.), a 4-h micropanel using single-substrate enzymatic test reactions, was compared with that of the API 20C AUX Clinical Yeast System (bioMerieux Vitek, Hazelwood, Mo.), a 48- to 72-h carbohydrate assimilation panel. Two hundred twenty-five yeasts, yeast-like fungi, and algae, comprising 28 species and including 30 isolates of Cryptococcus neoformans, an important pathogen not tested in appreciable numbers in other comparisons, were tested by both methods. On initial testing, 196 (87.1%) and 215 (95.6%) isolates were correctly identified by the RapID and API systems, respectively. Upon repeat testing, the number of correctly identified isolates increased to 220 (97.8%) for the RapID system and 223 (99.1%) for the API system. Reducing the turbidity of the test inoculum to that of a no. 3 McFarland turbidity standard, which is below that recommended by the manufacturer, resulted in the correct identification of most of the isolates initially misidentified by the RapID system, including 10 of 30 C. neoformans isolates. Concordance between the RapID and API results after repeat testing was 97.3%.

  9. Evaluation of a monoclonal antibody to ras peptide, RAP-5, claimed to bind preferentially to cells of infiltrating carcinomas.

    PubMed Central

    Robinson, A.; Williams, A. R.; Piris, J.; Spandidos, D. A.; Wyllie, A. H.

    1986-01-01

    RAP-5, a monoclonal antibody raised against a p21ras peptide, has been claimed to show immunohistochemical localisation of cells with infiltrative properties in human tumours. We confirmed that this antibody reveals pronounced cellular heterogeneity in human colonic neoplasms but could find no obvious relationship to infiltrative activity. RAP-5 bound to many different cell types, neoplastic and normal. In order to clarify the specificities of RAP-5 we applied it to two cell lines: nontumorigenic hamster fibroblasts in which ras expression is barely detectable, and a vigorously tumorigenic line derived from these fibroblasts by insertion of the human mutated Ha-ras oncogene in a high expression vector. Another antibody to p21ras, Y13-259, clearly distinguished between these cell lines both on immunoblots and immunocytochemically, but RAP-5 did not. Rather, it bound to proteins of a variety of molecular weights in both cell lines. The results show that RAP-5 is unlikely to be a useful reagent for detection of ras associated proteins in human tissues. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:3542007

  10. The yeast telomere-binding protein RAP1 binds to and promotes the formation of DNA quadruplexes in telomeric DNA.

    PubMed Central

    Giraldo, R; Rhodes, D

    1994-01-01

    The protein RAP1 is essential for the maintenance of the telomeres of Saccharomyces cerevisiae and binds in vitro to multiple sites found within the TG1-3 telomeric repeats. We show here that, in addition to its known binding activity for double-stranded DNA, RAP1 binds sequence-specifically to the GT-strands. This indicates that RAP1 is the protein that binds to the telomeric terminal GT-tails. Furthermore, we have found that RAP1 binds to and promotes the formation of G-tetrads, i.e. DNA quadruplexes, in GT-strand oligonucleotides at nanomolar concentrations. The formation of DNA quadruplexes appears to involve the intermolecular association of GT-strands. The minimal DNA-binding domain of RAP1 (DBD) binds only to double-stranded DNA, so that the novel DNA-binding activity we have found involves regions of the protein located outside of the DBD. The finding that a telomeric protein promotes the formation of G-tetrads argues for the use of DNA quadruplexes in telomere association. Images PMID:8194531

  11. 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.

  12. 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

  13. Voltage-gated ion channel Kv4.3 is associated with Rap guanine nucleotide exchange factors and regulates angiotensin receptor type 1 signaling to small G-protein Rap.

    PubMed

    Potapova, Irina A; Cohen, Ira S; Doronin, Sergey V

    2007-09-01

    The voltage-gated potassium channel Kv4.3 was coexpressed with its beta-subunit Kv channel-interacting protein 2 and the angiotensin type 1 receptor in HEK-293 cells. Proteomic analysis of proteins coimmunoprecipitated with Kv4.3 revealed that Kv4.3 is associated with Rap guanine nucleotide exchange factors MR-GEF and EPAC-1. Previously, we demonstrated that Kv4.3 interacts with the angiotensin type 1 receptor in HE293 cells and cardiac myocytes. On the basis of this, we investigated the angiotensin type 1 receptor signaling to small G-proteins Ras and Rap-1 in the presence and absence of the Kv4.3-Kv channel-interacting protein 2 macromolecular complex. Ras activation was not significantly affected by coexpression of Kv4.3 and Kv channel-interacting protein 2. Ras exhibited a rapid activation-inactivation pattern with maximum activity at 2.5 min after addition of angiotensin II. In contrast, activation of Rap-1 was affected dramatically by coexpression of Kv4.3 and Kv channel-interacting protein 2 with the angiotensin type 1 receptor. In the absence of Kv4.3 and Kv channel-interacting protein 2, stimulation of the angiotensin type 1 receptor resulted in steady activation of Rap-1 that reached a plateau 25 min after addition of angiotensin II. In the presence of Kv4.3 and Kv channel-interacting protein 2, Rap-1 reaches a maximum activity 2.5 min after addition of angiotensin II and then deactivates rapidly, demonstrating a pattern of activation similar to that of Ras. Our findings show that Kv4.3 regulates angiotensin type 1 receptor signaling to the small G-protein Rap-1.

  14. PERFORMANCE OF THE RAPS4/RAPS4-QF FOR DSM-5 COMPARED TO DSM-IV ALCOHOL USE DISORDERS IN THE GENERAL POPULATION: DATA FROM THE 2000–2010 NATIONAL ALCOHOL SURVEYS

    PubMed Central

    Cherpitel, Cheryl J.; Ye, Yu

    2015-01-01

    Background A number of relatively short screening instruments have been developed for identifying alcohol use disorders (AUD), but performance has been evaluated against the standard Diagnostic and Statistical Manual of Mental and Behavior Disorders (DSM) criteria, and it is not known how screening instruments may perform based on the newly formulated DSM-5 criteria, which is a radical departure from previous versions of the DSM. Analyzed here is the performance of the RAPS4/RAPS4-QF against DSM-5 criteria for AUD compared to DSM-IV dependence and abuse criteria. Methods Sensitivity and specificity are analyzed in a merged sample of 21,386 respondents from three National Alcohol Surveys of the U.S. general population (2000, 2005, 2010) Results Sensitivity of the RAPS4 was lower for DSM-5 AUD (62.5%) than for DSM-IV dependence (88%), while the RAPS4-QF was higher for DSM-5 AUD (90.3%) than for DSM-IV abuse (81.3%), or abuse/dependence (85.8%), while maintaining good specificity (84%). Sensitivity of the RAPS4-QF was higher for males (92%) compared to females (86.6%) and highest for whites (93.8%) followed by Hispanics (84.2%) and blacks (82.4%). Conclusions Screening instruments may not perform similarly for DSM-5 as for DSM-IV AUD, and data here suggest the RAPS4-QF may be a good instrument choice for identifying those meeting criteria for DSM-5 AUD. These data also suggest the need for additional research and a similar evaluation of other commonly used screening instruments for DSM-5 AUD. PMID:25823905

  15. 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.

  16. In vitro expression and redistribution of nucleolar proteins following the treatment 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

    2009-01-01

    In this study, we used a newly synthesized antitumor complex [RuLCl2]H.4H2O (RAP), having the same antitumor effects as cisplatin but showing lower cytotoxicity. We found that RAP-DNA adducts induce a high expression of proteins with high molecular weight and a low expression of proteins with low molecular weight. We choose two proteins: the upstream binding factor (UBF), an RNA polymerase I-specific transcription factor that recognizes the ribosomal RNA gene promoter and initiates transcription; and fibrillarin, which is involved in many posttranscriptional processes including pre-rRNA processing, pre-rRNA methylation, and ribosome assembly. Our results showed that UBF was present in high quantities in TG cell extracts treated with RAP with a major abundance of UBF1 more than UBF2, which was explained by a high affinity of UBF1 for DNA modified by RAP than UBF2; while fibrillarin was present in low quantities in protein extracts treated with RAP. Also, following treatment with RAP, there was a similar redistribution of UBF along the nucleus of TG cells as in the controls but with the presence of higher quantities of this factor in the nucleoplasm, which could be explained by an increase of the UBF affinity for the no nucleolar chromatin as a consequence of the modifications induced by RAP. Fibrillarin was found in low quantities in the fibrillar centers and in the nucleoplasm after treatment with RAP.

  17. 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…

  18. 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 Central

    Rösch, Thomas C.; Graumann, Peter L.

    2015-01-01

    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. PMID:26112413

  19. 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.

  20. 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.

  1. Retrospective study of long-term outcome after brain arteriovenous malformation rupture: the RAP score.

    PubMed

    Shotar, Eimad; Debarre, Matthieu; Sourour, Nader-Antoine; Di Maria, Federico; Gabrieli, Joseph; Nouet, Aurélien; Chiras, Jacques; Degos, Vincent; Clarençon, Frédéric

    2017-01-20

    OBJECTIVE The authors aimed to design a score for stratifying patients with brain arteriovenous malformation (BAVM) rupture, based on the likelihood of a poor long-term neurological outcome. METHODS The records of consecutive patients with BAVM hemorrhagic events who had been admitted over a period of 11 years were retrospectively reviewed. Independent predictors of a poor long-term outcome (modified Rankin Scale score ≥ 3) beyond 1 year after admission were identified. A risk stratification scale was developed and compared with the intracranial hemorrhage (ICH) score to predict poor outcome and inpatient mortality. RESULTS One hundred thirty-five patients with 139 independent hemorrhagic events related to BAVM rupture were included in this analysis. Multivariate logistic regression followed by stepwise analysis showed that consciousness level according to the Glasgow Coma Scale (OR 6.5, 95% CI 3.1-13.7, p < 10(-3)), hematoma volume (OR 1.8, 95% CI 1.2-2.8, p = 0.005), and intraventricular hemorrhage (OR 7.5, 95% CI 2.66-21, p < 10(-3)) were independently associated with a poor outcome. A 12-point scale for ruptured BAVM prognostication was constructed combining these 3 factors. The score obtained using this new scale, the ruptured AVM prognostic (RAP) score, was a stronger predictor of a poor long-term outcome (area under the receiver operating characteristic curve [AUC] 0.87, 95% CI 0.8-0.92, p = 0.009) and inpatient mortality (AUC 0.91, 95% CI 0.85-0.95, p = 0.006) than the ICH score. For a RAP score ≥ 6, sensitivity and specificity for predicting poor outcome were 76.8% (95% CI 63.6-87) and 90.8% (95% CI 81.9-96.2), respectively. CONCLUSIONS The authors propose a new admission score, the RAP score, dedicated to stratifying the risk of poor long-term outcome after BAVM rupture. This easy-to-use scoring system may help to improve communication between health care providers and consistency in clinical research. Only external prospective cohorts and population

  2. Essential role of Rap signal in pre-TCR-mediated beta-selection checkpoint in alphabeta T-cell development.

    PubMed

    Kometani, Kohei; Moriyama, Masaki; Nakashima, Yasuhiro; Katayama, Yoshinori; Wang, Shu-Fang; Yamasaki, Sho; Saito, Takashi; Hattori, Masakazu; Minato, Nagahiro

    2008-12-01

    We demonstrate that lck promoter-driven conditional expression of transgenic SPA-1, a Rap GTPase-activation protein, causes a profound defect of alphabeta T-cell development at the CD4/CD8 double-negative (DN) stage due to enhanced cell death without affecting gammadelta T-cell development. The effect was specific to the DN stage, because CD4 promoter-driven SPA-1 expression hardly affected T-cell development. Rap1A17, a dominant-negative Rap mutant, interfered with the generation of double-positive (DP) cells from Rag2(-/-) fetal thymocytes in vitro in the presence of anti-CD3epsilon antibody and Notch ligand. Rap GTPases were activated in a DN cell line by the expression of self-oligomerizing CD3 (CD8:CD3epsilon chimera), which substituted autonomous pre-T-cell receptor (TCR) signal, inducing CD69 expression and CD25 down-regulation. Reciprocally, expression of C3G, a Rap guanine nucleotide exchange factor, in both normal and Rag2(-/-) DN cells markedly enhanced Notch-dependent generation and expansion of DP cells without additional anti-CD3epsilon antibody, thus bypassing pre-TCR. Defective alphabeta T-cell development in the conditional SPA-1-transgenic mice was restored completely by introducing a p53(-/-) mutation. These results suggest that endogenous Rap GTPases downstream of pre-TCR play an essential role in rescuing pre-T cells from the p53-mediated checkpoint response, thus allowing Notch-mediated expansion and differentiation.

  3. 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

  4. In Vitro Assay for the Rap GTPase-Activating Protein Activity of the Purified Cytoplasmic Domain of Plexin.

    PubMed

    Pascoe, Heath G; Wang, Yuxiao; Zhang, Xuewu

    2017-01-01

    Plexins are cell surface receptors that bind semaphorins and regulate essential processes such as axon guidance and angiogenesis. The cytoplasmic regions of plexins contain a functionally essential GTPase-activating protein (GAP) domain, which initiates downstream signaling by specifically inactivating the Rap GTPase. Here we describe the methods for expression and purification of the plexin cytoplasmic region in E. coli, and characterization of its GAP activity using a photometric assay. We also provide a protocol for measuring GAP activity of single-chain constructs with Rap covalently linked to the plexin cytoplasmic region.

  5. Protein Kinase A-independent Ras Protein Activation Cooperates with Rap1 Protein to Mediate Activation of the Extracellular Signal-regulated Kinases (ERK) by cAMP.

    PubMed

    Li, Yanping; Dillon, Tara J; Takahashi, Maho; Earley, Keith T; Stork, Philip J S

    2016-10-07

    Cyclic adenosine monophosphate (cAMP) is an important mediator of hormonal stimulation of cell growth and differentiation through its activation of the extracellular signal-regulated kinase (ERK) cascade. Two small G proteins, Ras and Rap1, have been proposed to mediate this activation, with either Ras or Rap1 acting in distinct cell types. Using Hek293 cells, we show that both Ras and Rap1 are required for cAMP signaling to ERKs. The roles of Ras and Rap1 were distinguished by their mechanism of activation, dependence on the cAMP-dependent protein kinase (PKA), and the magnitude and kinetics of their effects on ERKs. Ras was required for the early portion of ERK activation by cAMP and was activated independently of PKA. Ras activation required the Ras/Rap guanine nucleotide exchange factor (GEF) PDZ-GEF1. Importantly, this action of PDZ-GEF1 was disrupted by mutation within its putative cyclic nucleotide-binding domain within PDZ-GEF1. Compared with Ras, Rap1 activation of ERKs was of longer duration. Rap1 activation was dependent on PKA and required Src family kinases and the Rap1 exchanger C3G. This is the first report of a mechanism for the cooperative actions of Ras and Rap1 in cAMP activation of ERKs. One physiological role for the sustained activation of ERKs is the transcription and stabilization of a range of transcription factors, including c-FOS. We show that the induction of c-FOS by cAMP required both the early and sustained phases of ERK activation, requiring Ras and Rap1, as well as for each of the Raf isoforms, B-Raf and C-Raf.

  6. 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

  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. Enzyme profiles of oral spirochetes in RapID-ANA system.

    PubMed Central

    Syed, S A; Salvador, S L; Loesche, W J

    1988-01-01

    Enzyme profiles of oral Treponema species were determined by using RapID-ANA (Innovative Diagnostic System, Atlanta, Ga.), a 4-h test system which detects 18 enzymatic reactions, including aminopeptidases and glycosidases. Seventy-two clinical isolates of Treponema denticola, four reference strains of T. denticola (ATCC 35404, ATCC 35405, ATCC 35520, and ATCC 33521), one strain of T. vincentii (ATCC 35580), and two strains of T. socranskii subspecies (T. socranskii subsp. buccale ATCC 35534 and T. socranskii subsp. socranskii ATCC 35536) were used in this study. All T. denticola strains produced indole and a variety of aminopeptidases and glycosidases. These organisms could be differentiated into two groups on the basis of tetrazolium reductase and serine, phenylalanine, and glycine aminopeptidase activities. T. vincentii produced N-acetylglucosaminidase and arginine aminopeptidase, which facilitated the differentiation of this organism from T. socranskii subspecies and the T. denticola group. T. socranskii subspecies gave positive reactions for alkaline phosphatase only. These findings suggest that the RapID-ANA system is useful for enzymatic characterization and differentiation of oral spirochetes. PMID:3183013

  9. RAP: a computer program for exploring similarities in behavior sequences using random projections.

    PubMed

    Quera, Vicenç

    2008-02-01

    A computer program (RAP, for random projection) for exploring similarities between and within sequences of behavior is presented. Given a time window of a sequence, the program calculates a signature, a real-valued vector that is a random projection of the contents of the window (i.e., the codes occurring within it and their relative location, or onset and offset times) into an arbitrary K-dimensional space. Then, given two different time windows from the same sequence or from different sequences, their similarity is computed as an inverse function of the Euclidean distance between their respective signatures. By defining moving (overlapped or not overlapped) windows along each sequence and calculating similarities between every pair of windows from the two sequences, a map of similarities or possible recurrent patterns is obtained; the RAP program represents them as gray-level lattices, which are displayed as mouse-sensitive images in an HTML file. Computation of similarities is based on the random projection method, as presented by Mannila and Seppänen (2001), for the analysis of sequences of events. The program reads sequence data files in Sequential Data Interchange Standard (SDIS) format (Bakeman Quera, 1992,1995a).

  10. Sumoylation stabilizes RACK1B and enhance its interaction with RAP2.6 in the abscisic acid response

    PubMed Central

    Guo, Rongkai; Sun, Weining

    2017-01-01

    The highly conserved eukaryotic WD40 repeat protein, Receptor for Activated C Kinase 1 (RACK1), is involved in the abscisic acid (ABA) response in Arabidopsis. However, the regulation of RACK1 and the proteins with which it interacts are poorly understood. Here, we show that RACK1B is sumoylated at four residues, Lys50, Lys276, Lys281 and Lys291. Sumoylation increases RACK1B stability and its tolerance to ubiquitination-mediated degradation in ABA response. As a result, sumoylation leads to enhanced interaction between RACK1B and RAP2.6, an AP2/ERF family transcription factor. RACK1B binds directly to the AP2 domain of RAP2.6, which alters the affinity of RAP2.6 for CE1 and GCC cis-acting regulatory elements. Taken together, our findings illustrate that protein stability controlled by dynamic post-transcriptional modification is a critical regulatory mechanism for RACK1B, which functions as scaffold protein for RAP2.6 in ABA signaling. PMID:28272518

  11. 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…

  12. 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.

  13. 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…

  14. Novel mechanisms of controlling the activities of the transcription factors Spo0A and ComA by the plasmid-encoded quorum sensing regulators Rap60-Phr60 in Bacillus subtilis

    PubMed Central

    Boguslawski, Kristina M.; Hill, Patrick A.; Griffith, Kevin L.

    2015-01-01

    Summary Bacillus subtilis and its closest relatives have multiple rap-phr quorum sensing gene pairs that coordinate a variety of physiological processes with population density. Extra-chromosomal rap-phr genes are also present on mobile genetic elements, yet relatively little is known about their function. In this work, we demonstrate that Rap60-Phr60 from plasmid pTA1060 coordinates a variety of biological processes with population density including sporulation, cannibalism, biofilm formation and genetic competence. Similar to other Rap proteins that control sporulation, Rap60 modulates phosphorylation of the transcription factor Spo0A by acting as a phosphatase of Spo0F~P, an intermediate of the sporulation phosphorelay system. Additionally, Rap60 plays a noncanonical role in regulating the autophosphorylation of the sporulation-specific kinase KinA, a novel activity for Rap proteins. In contrast, Rap proteins that modulate genetic competence interfere with DNA binding by the transcription factor ComA. Rap60 regulates the activity of ComA in a unique manner by forming a Rap60–ComA–DNA ternary complex that inhibits transcription of target genes. Taken together, this work provides new insight into two novel mechanisms of regulating Spo0A and ComA by Rap60 and expands our general understanding of how plasmid-encoded quorum sensing pairs regulate important biological processes. PMID:25598361

  15. Regulation of glia number in Drosophila by Rap/Fzr, an activator of the anaphase-promoting complex, and Loco, an RGS protein.

    PubMed

    Kaplow, Margarita E; Korayem, Adam H; Venkatesh, Tadmiri R

    2008-04-01

    Glia mediate a vast array of cellular processes and are critical for nervous system development and function. Despite their immense importance in neurobiology, glia remain understudied and the molecular mechanisms that direct their differentiation are poorly understood. Rap/Fzr is the Drosophila homolog of the mammalian Cdh1, a regulatory subunit of the anaphase-promoting complex/cyclosome (APC/C). APC/C is an E3 ubiquitin ligase complex well characterized for its role in cell cycle progression. In this study, we have uncovered a novel cellular role for Rap/Fzr. Loss of rap/fzr function leads to a marked increase in the number of glia in the nervous system of third instar larvae. Conversely, ectopic expression of UAS-rap/fzr, driven by repo-GAL4, results in the drastic reduction of glia. Data from clonal analyses using the MARCM technique show that Rap/Fzr regulates the differentiation of surface glia in the developing larval nervous system. Our genetic and biochemical data further indicate that Rap/Fzr regulates glial differentiation through its interaction with Loco, a regulator of G-protein signaling (RGS) protein and a known effector of glia specification. We propose that Rap/Fzr targets Loco for ubiquitination, thereby regulating glial differentiation in the developing nervous system.

  16. 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

    2012-08-09

    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.

  17. The de-ubiquitylating enzymes USP26 and USP37 regulate homologous recombination by counteracting RAP80

    PubMed Central

    Typas, Dimitris; Luijsterburg, Martijn S.; Wiegant, Wouter W.; Diakatou, Michaela; Helfricht, Angela; Thijssen, Peter E.; van de Broek, Bram; Mullenders, Leon H.; van Attikum, Haico

    2015-01-01

    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

  18. 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.

  19. Neural correlates of lyrical improvisation: an FMRI study of freestyle rap.

    PubMed

    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.

  20. The challenge of treating conduct disorder in low-resourced settings: rap music to the rescue.

    PubMed

    Evans, Dylan J

    2010-12-01

    Conduct disorder is one of the most frequent serious childhood problems that present for treatment in community clinic settings. Evidence-based treatments for conduct disorder are intensive and require considerable resources to implement. In low-resourced contexts it is often not feasible to implement evidence-based treatments in their current form, which poses significant challenges for clinicians attempting to treat children in these settings. This article explores these challenges using a case study of the treatment of a young adolescent boy with a short-term multisystem intervention where rap music was employed as a powerful tool to facilitate an empathic connection in therapy and as a projective technique to explore underlying emotional difficulties.

  1. 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

  2. DNA DSB repair pathway choice: an orchestrated handover mechanism.

    PubMed

    Kakarougkas, A; Jeggo, P A

    2014-03-01

    DNA double strand breaks (DSBs) are potential lethal lesions but can also lead to chromosome rearrangements, a step promoting carcinogenesis. DNA non-homologous end-joining (NHEJ) is the major DSB rejoining process and occurs in all cell cycle stages. Homologous recombination (HR) can additionally function to repair irradiation-induced two-ended DSBs in G2 phase. In mammalian cells, HR predominantly uses a sister chromatid as a template for DSB repair; thus HR functions only in late S/G2 phase. Here, we review current insight into the interplay between HR and NHEJ in G2 phase. We argue that NHEJ represents the first choice pathway, repairing approximately 80% of X-ray-induced DSBs with rapid kinetics. However, a subset of DSBs undergoes end resection and repair by HR. 53BP1 restricts resection, thereby promoting NHEJ. During the switch from NHEJ to HR, 53BP1 is repositioned to the periphery of enlarged irradiation-induced foci (IRIF) via a BRCA1-dependent process. K63-linked ubiquitin chains, which also form at IRIF, are also repositioned as well as receptor-associated protein 80 (RAP80), a ubiquitin binding protein. RAP80 repositioning requires POH1, a proteasome component. Thus, the interfacing barriers to HR, 53BP1 and RAP80 are relieved by POH1 and BRCA1, respectively. Removal of RAP80 from the IRIF core is required for loss of the ubiquitin chains and 53BP1, and for efficient replication protein A foci formation. We propose that NHEJ is used preferentially to HR because it is a compact process that does not necessitate extensive chromatin changes in the DSB vicinity.

  3. 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.

  4. 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.

  5. 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.

  6. 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.'

  7. The rhizobial adhesion protein RapA1 is involved in adsorption of rhizobia to plant roots but not in nodulation.

    PubMed

    Mongiardini, Elías J; Ausmees, Nora; Pérez-Giménez, Julieta; Julia Althabegoiti, María; Ignacio Quelas, Juan; López-García, Silvina L; Lodeiro, Aníbal R

    2008-08-01

    The effect of the rhizobium adhesion protein RapA1 on Rhizobium leguminosarum bv. trifolii adsorption to Trifolium pratense (red clover) roots was investigated. We altered RapA1 production by cloning its encoding gene under the plac promoter into the stable vector pHC60. After introducing this plasmid in R. leguminosarum bv. trifolii, three to four times more RapA1 was produced, and two to five times higher adsorption to red clover roots was obtained, as compared with results for the empty vector. Enhanced adsorption was also observed on soybean and alfalfa roots, not related to R. leguminosarum cross inoculation groups. Although the presence of 1 mM Ca2+ during rhizobial growth enhanced adsorption, it was unrelated to RapA1 level. Similar effects were obtained when the same plasmid was introduced in Rhizobium etli for its adsorption to bean roots. Although root colonization by the RapA1-overproducing strain was also higher, nodulation was not enhanced. In addition, in vitro biofilm formation was similar to the wild-type both on polar and on hydrophobic surfaces. These results suggest that RapA1 receptors are present in root but not on inert surfaces, and that the function of this protein is related to rhizosphere colonization.

  8. AgrAbility Project

    MedlinePlus

    ... It’s About Hope AgrAbility on Twitter AgrAbility on Facebook AgrAbility on You Tube AgrAbility… It’s About Hope ... summary report available... AgrAbility Harvest Get a copy Facebook Posts National AgrAbility Project 12 hours ago Good ...

  9. 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.

  10. TLR signaling paralyzes monocyte chemotaxis through synergized effects of p38 MAPK and global Rap-1 activation.

    PubMed

    Yi, Ling; Chandrasekaran, Prabha; Venkatesan, Sundararajan

    2012-01-01

    Toll-like receptors (TLRs) that recognize pathogen associated molecular patterns and chemoattractant receptors (CKRs) that orchestrate leukocyte migration to infected tissue are two arms of host innate immunity. Although TLR signaling induces synthesis and secretion of proinflammatory cytokines and chemokines, which recruit leukocytes, many studies have reported the paradoxical observation that TLR stimulation inhibits leukocyte chemotaxis in vitro and impairs their recruitment to tissues during sepsis. There is consensus that physical loss of chemokine receptor (CKR) at the RNA or protein level or receptor usage switching are the mechanisms underlying this effect. We show here that a brief (<15 min) stimulation with LPS (lipopolysaccharide) at ~0.2 ng/ml inhibited chemotactic response from CCR2, CXCR4 and FPR receptors in monocytes without downmodulation of receptors. A 3 min LPS pre-treatment abolished the polarized accumulation of F-actin, integrins and PIP(3) (phosphatidylinositol-3,4,5-trisphosphate) in response to chemokines in monocytes, but not in polymorphonuclear neutrophils (PMNs). If chemoattractants were added before or simultaneously with LPS, chemotactic polarization was preserved. LPS did not alter the initial G-protein signaling, or endocytosis kinetics of agonist-occupied chemoattractant receptors (CKRs). The chemotaxis arrest did not result from downmodulation of receptors or from inordinate increase in adhesion. LPS induced rapid p38 MAPK activation, global redistribution of activated Rap1 (Ras-proximate-1 or Ras-related protein 1) GTPase and Rap1GEF (guanylate exchange factor) Epac1 (exchange proteins activated by cyclic AMP) and disruption of intracellular gradient. Co-inhibition of p38 MAPK and Rap1 GTPase reversed the LPS induced breakdown of chemotaxis suggesting that LPS effect requires the combined function of p38 MAPK and Rap1 GTPase.

  11. Usefulness of CDK5RAP3, CCNB2, and RAGE genes for the diagnosis of lung adenocarcinoma.

    PubMed

    Stav, D; Bar, I; Sandbank, J

    2007-01-01

    We used oligonucleotide microarrays with probe sets to 22,283 genes to analyze the gene expression profile of lung adenocarcinoma. Cancerous and noncancerous tissue samples were obtained from 23 patients with stage I or II lung cancer; 18 tissue pairs and 5 cancerous tissues. A list of 2065 genes that differentiate between cancerous and noncancerous tissues was generated using Winsorized paired t-tests. We analyzed CDK5RAP3 and CCNB2, which are involved in cell cycle progression, and RAGE. The first 2 of these 3 genes proved to be overexpressed in tumor tissue, whereas the RAGE gene was suppressed in tumor tissue. When CDK5RAP3 and CCNB2 were examined in individual patients we found that in cases where one of these genes was only slightly overexpressed the other was highly overexpressed. The combined expression of the 2 cell cycle genes was found to be statistically significant for differentiating between cancerous and noncancerous tissues. Inclusion of the data for the RAGE gene made the differentiation more powerful. The gene expression ratio gave a clear result: when CDK5RAP3 was expressed more than RAGE, the tissue was carcinomatous, and vice versa. We therefore conclude that these 3 genes may be used as a very reliable biomarker of lung adenocarcinoma.

  12. 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

  13. Function of wild-type or mutant Rac2 and Rap1a GTPases in differentiated HL60 cell NADPH oxidase activation.

    PubMed

    Gabig, T G; Crean, C D; Mantel, P L; Rosli, R

    1995-02-01

    Studies of neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation in a cell-free system showed that the low molecular-weight guanosine triphosphatase (GTPase) Rac was required, and that Rap1a may participate in activation of the catalytic complex. Full-length posttranslationally modified Rac2 was active, whereas only the 1-166 truncated form of Rap1a was functional in the cell-free system, and thus, clarification of the function of Rap1a and Rac2 in intact human phagocytes is needed to provide further insight into their roles as signal transducers from plasma membrane receptors. In the present studies, oligonucleotide-directed mutagenesis was used to introduce a series of mutations into human rap1a or rac2 in the mammalian expression vector pSR alpha neo. HL60 cells transfected with wild-type or mutated rac2 or rap1a cDNA constructs and control HL60 cells transfected with the pSR alpha neo vector containing no inserted cDNA were selected in G418-containing media, then subclones were isolated. Compared with the parent HL60 cells, each of the stable transfected cell lines differentiated similarly into neutrophil-like cells and expressed comparable levels of NADPH oxidase components p47-phox, p67-phox and gp91-phox. The differentiated vector control cell line produced O2. in response to receptor stimulation at rates that were not significantly different from parent HL60 cells. O2-. production by differentiated cell lines expressing mutated N17 Rap1a or N17 Rac2 dominant-negative proteins was inhibited, whereas O2-. production by the subline overexpressing wild-type Rap1a was increased by fourfold. O2-. production by the differentiated cell line expressing GTPase-defective V12 Rap1a was also significantly inhibited, a finding that is consistent with a requirement for cycling between guanosine diphosphate- and GTP-bound forms of Rap1a for continuous NADPH oxidase activation in intact neutrophils. A model is proposed in which Rac2 mediates

  14. Conceptions of Ability.

    ERIC Educational Resources Information Center

    Jagacinski, Carolyn M.; Nicholls, John G.

    Two different conceptions of ability are proposed. The first conception of ability is more differentiated and generally employed by adults and older children. Here ability level is defined with reference to the performance of others assuming that optimum effort was employed. High ability means higher than others. The second conception of ability…

  15. 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.

  16. 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.

  17. miR-28-5p acts as a tumor suppressor in renal cell carcinoma for multiple antitumor effects by targeting RAP1B

    PubMed Central

    Ding, Meng; Zhong, Jinsha; Zhang, Chen-Yu; Ge, Jingping; Wang, Junjun; Zhang, Chunni

    2016-01-01

    The incidence and mortality rate of renal cell carcinoma (RCC) have been significantly increasing; however, the mechanisms involved in RCC development and progression are unclear. In this study, we found that miR-28-5p was decreased in RCC tumor specimens and several renal carcinoma cell lines. By using a combination of luciferase reporter assays and western blotting, we identified RAP1B, a Ras-related small GTP-binding oncoprotein implicated in a variety of tumors, as a direct target of miR-28-5p in RCC. The RAP1B protein level was increased in RCC tumor specimens and renal carcinoma cell lines, and this was inversely correlated with miR-28-5p expression. In vitro gain-of-function and loss-of-function studies in human renal carcinoma cell lines, demonstrated that miR-28-5p suppressed cell proliferation and migration by directly inhibiting RAP1B, and this effect was reversed by co-transfection with RAP1B. In addition, the stable overexpression of miR-28-5p inhibited tumor cell proliferation in vivo. This newly identified miR-28-5p/RAP1B axis provides a novel mechanism for the pathogenesis of RCC, and molecules in this axis may serve as potential biomarkers and therapeutic targets for RCC. PMID:27729617

  18. Indictment of Ability Grouping

    ERIC Educational Resources Information Center

    Tuckman, Bruce

    1972-01-01

    The use of ability grouping restricts students to interact with others who have been identified as similar in ability and carries with it the stigma of failure and the operation of the self-fulfilling prophecy. (Author)

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. 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)

  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, 2012 CFR

    2012-07-01

    ... assumed to be in compliance with this requirement. (e) These alternative locations are remediation waste... 40 Protection of Environment 28 2012-07-01 2012-07-01 false May I perform remediation waste management activities under a RAP at a location removed from the area where the remediation wastes...

  5. 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.

  6. 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…

  7. Localization and role of RAP55/LSM14 in HeLa cells: a new finding on the mitotic spindle assembly.

    PubMed

    Mili, Donia; Georgesse, Dane; Kenani, Abderraouf

    2015-01-01

    The MAP family includes large proteins like MAP-1A, MAP-1B, MAP-1C, MAP-2, and MAP-4 and smaller components like tau and MAP-2C. This article focuses on the relevant aspects of RAP55/LSM14 position with emphasis on its role in mitotic spindle formation and stability. In this context, the localization of RNA associated Protein 55kDa (RAP55/LSM14) during mitosis was identified as a Mitotic Spindle Protein (MSP). We found a new location obtained by expressing GFP-tagged proteins in HeLa Cells during mitosis that has never been previously reported. We demonstrated also, for the first time, that the depletion of RAP55/LSM14 destabilizes spindle assembly, stops cells in mitosis and induces many other cell cytoskeletal disorders. Finally, by using an "in vitro" assay investigation, we found that RAP55/LSM14 binds directly the tubulin and that is implicated in the process of the mitotic spindle stabilization, which is a novel discovery in this field of research.

  8. Decreased interactions of mutant muscle LIM protein (MLP) with N-RAP and alpha-actinin and their implication for hypertrophic cardiomyopathy.

    PubMed

    Gehmlich, Katja; Geier, Christian; Osterziel, Karl Josef; Van der Ven, Peter F M; Fürst, Dieter O

    2004-08-01

    Previous work has shown that mutations in muscle LIM protein (MLP) can cause hypertrophic cardiomyopathy (HCM). In order to gain an insight into the molecular basis of the disease phenotype, we analysed the binding characteristics of wild-type MLP and of the (C58G) mutant MLP that causes hypertrophic cardiomyopathy. We show that MLP can form a ternary complex with two of its previously documented myofibrillar ligand proteins, N-RAP and alpha-actinin, which indicates the presence of distinct, non-overlapping binding sites. Our data also show that, in comparison to wild-type MLP, the capacity of the mutated MLP protein to bind both N-RAP and alpha-actinin is significantly decreased. In addition, this single point mutation prevents zinc coordination and proper folding of the second zinc-finger in the first LIM domain, which consequently renders the protein less stable and more susceptible to proteolysis. The molecular basis for HCM-causing mutations in the MLP gene might therefore be an alteration in the equilibrium of interactions of the ternary complex MLP-N-RAP-alpha-actinin. This assumption is supported by the previous observation that in the pathological situation accompanied by MLP down regulation, cardiomyocytes try to compensate for the decreased stability of MLP protein by increasing the expression of its ligand N-RAP, which might finally result in the development of myocyte disarray that is characteristic of this disease.

  9. 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

  10. 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.

  11. 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.

  12. Microstructure evolution and thixoforming behavior of 7075 aluminum alloy in the semi-solid state prepared by RAP method

    NASA Astrophysics Data System (ADS)

    Fu, Jin-long; Wang, Kai-kun; Li, Xiao-wei; Zhang, Hai-kuan

    2016-12-01

    The effects of isothermal treatments on the microstructural evolution and coarsening rate of semi-solid 7075 aluminum alloy produced via the recrystallization and partial remelting (RAP) process were investigated. Samples of 7075 aluminum alloy were subjected to cold extrusion, and semi-solid treatment was carried out for 5-30 min at temperatures ranging from 580 to 605°C. A backward-extrusion experiment was conducted to investigate liquid segregation during the thixoforming process. The results revealed that obvious grain coarsening and spheroidization occurred during prolonged isothermal treatments. In addition, higher soaking temperatures promoted the spheroidization and coarsening process because of the increased liquid fraction and the melting of second phases. Segregation of the liquid phase caused by the difference in fluidity between the liquid and the solid phases was observed in different regions of the thixoformed specimens.

  13. Epac1 interacts with importin β1 and controls neurite outgrowth independently of cAMP and Rap1

    PubMed Central

    Baameur, Faiza; Singhmar, Pooja; Zhou, Yong; Hancock, John F.; Cheng, Xiaodong; Heijnen, Cobi J.; Kavelaars, Annemieke

    2016-01-01

    Exchange protein directly activated by cAMP-1 (Epac1) is a cAMP sensor that regulates multiple cellular functions including cellular migration, proliferation and differentiation. Classically, Epac1 is thought to exert its effects through binding of cAMP leading to a conformational change in Epac1 and its accumulation at the plasma membrane (PM) where it activates Rap1. In search for regulators of Epac1 activity, we show here that importin β1 (impβ1) is an Epac1 binding partner that prevents PM accumulation of Epac1. We demonstrate that in the absence of impβ1, endogenous as well as overexpressed Epac1 accumulate at the PM. Moreover, agonist-induced PM translocation of Epac1 leads to dissociation of Epac1 from impβ1. Localization of Epac1 at the PM in the absence of impβ1, requires residue R82 in its DEP domain. Notably, the PM accumulation of Epac1 in the absence of impβ1 does not require binding of cAMP to Epac1 and does not result in Rap1 activation. Functionally, PM accumulation of Epac1, an Epac1 mutant deficient in cAMP binding, or an Epac1 mutant tethered to the PM, is sufficient to inhibit neurite outgrowth. In conclusion, we uncover a cAMP-independent function of Epac1 at the PM and demonstrate that impβ1 controls subcellular localization of Epac1. PMID:27808165

  14. 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.

  15. Ability Is Ageless.

    ERIC Educational Resources Information Center

    Wooten, Andrea

    2002-01-01

    Experience Works is a national organization that provides training and employment services to older adults. Its Prime Time Awards Program honors contributions of older workers in their 70s and beyond, demonstrating the continued ability and productivity of this population as well as the benefits they derive from productive work. (SK)

  16. Computerized Adaptive Ability Measurement.

    ERIC Educational Resources Information Center

    Weiss, David J.

    The general objective of a research program on adaptive testing was to identify several sources of potential error in test scores, and to study adaptive testing as a means for reducing these errors. Errors can result from the mismatch of item difficulty to the individual's ability; the psychological effects of testing and the test environment; the…

  17. 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…

  18. Promoting Logical Ability

    ERIC Educational Resources Information Center

    Osborne, Alan R.

    1973-01-01

    This article reports one search for factors or conditions shaping the child's growth in logical ability. The search indicated the existence of a relationship between the quantity of teacher talk that contains the language of logic and the change exhibited by students. Implications for classroom practice are discussed. (JA)

  19. Conservatism and Cognitive Ability

    ERIC Educational Resources Information Center

    Stankov, Lazar

    2009-01-01

    Conservatism and cognitive ability are negatively correlated. The evidence is based on 1254 community college students and 1600 foreign students seeking entry to United States' universities. At the individual level of analysis, conservatism scores correlate negatively with SAT, Vocabulary, and Analogy test scores. At the national level of…

  20. Sequence heterogeneity in the gene encoding the rhoptry-associated protein-1 (RAP-1) of Babesia caballi isolates from South Africa.

    PubMed

    Bhoora, Raksha; Quan, Melvyn; Zweygarth, Erich; Guthrie, Alan J; Prinsloo, Sandra A; Collins, Nicola E

    2010-05-11

    A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) developed for the detection of antibody specific for Babesia caballi was used to test sera collected from 1237 South African horses. None of these samples tested positive using the cELISA, although 63 samples tested positive for B. caballi antibody using the indirect fluorescent antibody test (IFAT). We therefore characterized the rap-1 gene that codes for the antigen (rhoptry-associated protein, RAP-1) used in the cELISA, from South African B. caballi isolates. Three sets of primers were designed to amplify the complete gene and flanking regions (approximately 1800 bp), but only one set of primers yielded PCR products, and we were only able to amplify a region at the 5' end of the gene (615 bp) from ten South African B. caballiin vitro-cultured isolates. Sequence data from seven of these were obtained. The sequences showed between 79% and 81% identity to B. caballirap-1 gene sequences that have been reported in the literature (accession numbers: AF092736 and AB017700). The GenomeWalker Universal kit (Clonetech) was used to amplify the regions flanking the 615bp B. caballirap-1 fragment from two South African isolates. Amplified products were cloned into the pGEM-T Easy vector and sequenced. The complete rap-1 gene sequence, comprising a single open reading frame of 1479 bp that encodes a protein consisting of 493 amino acids, was obtained from the two South African isolates. This sequence data was used to redesign the amplification primers and rap-1 homologues were obtained from a further eight isolates. BLASTP analysis indicated an amino acid identity of between 57.9% and 65.1% to the two RAP-1 protein sequences, AF092736 and AB017700, with most differences occurring at the carboxy-terminus. The amino acid sequence differences probably explain why it was not possible to detect B. caballi antibody in IFAT positive sera from South Africa using the cELISA. Redesigning the current cELISA using a