Compensatory embryonic response to allele-specific inactivation of the murine X-linked gene Hcfc1.

Early in female mammalian embryonic development, cells randomly inactivate one of the two X chromosomes to achieve overall equal inactivation of parental X-linked alleles. Hcfc1 is a highly conserved X-linked mouse gene that encodes HCF-1 - a transcriptional co-regulator implicated in cell proliferation in tissue culture cells. By generating a Cre-recombinase inducible Hcfc1 knock-out (Hcfc1(lox)) allele in mice, we have probed the role of HCF-1 in actively proliferating embryonic cells and in cell-cycle re-entry of resting differentiated adult cells using a liver regeneration model. HCF-1 function is required for both extraembryonic and embryonic development. In heterozygous Hcfc1(lox/+) female embryos, however, embryonic epiblast-specific Cre-induced Hcfc1 deletion (creating an Hcfc1(epiKO) allele) around E5.5 is well tolerated; it leads to a mixture of HCF-1-positive and -negative epiblast cells owing to random X-chromosome inactivation of the wild-type or Hcfc1(epiKO) mutant allele. At E6.5 and E7.5, both HCF-1-positive and -negative epiblast cells proliferate, but gradually by E8.5, HCF-1-negative cells disappear owing to cell-cycle exit and apoptosis. Although generating a temporary developmental retardation, the loss of HCF-1-negative cells is tolerated, leading to viable heterozygous offspring with 100% skewed inactivation of the X-linked Hcfc1(epiKO) allele. In resting adult liver cells, the requirement for HCF-1 in cell proliferation was more evident as hepatocytes lacking HCF-1 fail to re-enter the cell cycle and thus to proliferate during liver regeneration. The survival of the heterozygous Hcfc1(epiKO/+) female embryos, even with half the cells genetically compromised, illustrates the developmental plasticity of the post-implantation mouse embryo - in this instance, permitting survival of females heterozygous for an X-linked embryonic lethal allele.

Modifications post-traductionnelles des canaux calciques cardiaques de type L : identification des résidus asparagine qui participent à la glycosylation de la sous-unité auxiliaire CaVα2δ1

Les canaux calciques de type L CaV1.2 sont principalement responsables de l’entrée des ions calcium pendant la phase plateau du potentiel d’action des cardiomyocytes ventriculaires. Cet influx calcique est requis pour initier la contraction du muscle cardiaque. Le canal CaV1.2 est un complexe oligomérique qui est composé de la sous-unité principale CaVα1 et des sous-unités auxiliaires CaVβ et CaVα2δ1. CaVβ joue un rôle déterminant dans l’adressage membranaire de la sous-unité CaVα1. CaVα2δ1 stabilise l’état ouvert du canal mais le mécanisme moléculaire responsable de cette modulation n’a pas été encore identifié. Nous avons récemment montré que cette modulation requiert une expression membranaire significative de CaVα2δ1 (Bourdin et al. 2015). CaVα2δ1 est une glycoprotéine qui possède 16 sites potentiels de glycosylation de type N. Nous avons donc évalué le rôle de la glycosylation de type-N dans l’adressage membranaire et la stabilité de CaVα2δ1. Nous avons d’abord confirmé que la protéine CaVα2δ1 recombinante, telle la protéine endogène, est significativement glycosylée puisque le traitement à la PNGase F se traduit par une diminution de 50 kDa de sa masse moléculaire, ce qui est compatible avec la présence de 16 sites Asn. Il s’est avéré par ailleurs que la mutation simultanée de 6/16 sites (6xNQ) est suffisante pour 1) réduire significativement la densité de surface de! CaVα2δ1 telle que mesurée par cytométrie en flux et par imagerie confocale 2) accélérer les cinétiques de dégradation telle qu’estimée après arrêt de la synthèse protéique et 3) diminuer la modulation fonctionnelle des courants générés par CaV1.2 telle qu’évaluée par la méthode du « patch-clamp ». Les effets les plus importants ont toutefois été obtenus avec les mutants N663Q, et les doubles mutants N348Q/N468Q, N348Q/N812Q, N468Q/N812Q. Ensemble, ces résultats montrent que Asn663 et à un moindre degré Asn348, Asn468 et Asn812 contribuent à la biogenèse et la stabilité de CaVα2δ1 et confirment que la glycosylation de type N de CaVα2δ1 est nécessaire à la fonction du canal calcique cardiaque de type L.

The regulation of integrin-linked kinase in human platelets: evidence for involvement in the regulation of integrin alpha(2)beta(1)

Background: Activation of the platelet integrin alpha(2)beta(1) is closely regulated due to the high thrombogenicity of its ligand. As a beta(1) interacting kinase, ILK represents a candidate intracellular regulator of alpha(2)beta(1) in human platelets. Objectives We investigated the regulation of ILK in human platelets and the role of ILK in regulating alpha(2)beta(1) activation in HEL cells, a megakaryocytic cell line. Methods: An in-vitro kinase assay was used to determine the effect of platelet agonists on ILK kinase activity together with the contribution of PI3K and PKC on ILK activation. Interaction of ILK with beta(1)-integrin subunits was investigated by coimmunoprecipitation and the role of ILK in regulating alpha(2)beta(1) function assessed by overexpression studies in HEL cells. Results: We report that collagen and thrombin modulate ILK kinase activity in human platelets in an aggregation-independent manner. Furthermore, ILK activity is dually regulated by PI3K and PKC in thrombin-stimulated platelets and regulated by PI3K in collagen-stimulated cells. ILK associates with the beta(1)-integrin subunits immunoprecipitated from platelet cell lysates, an association which increased upon collagen stimulation. Overexpression of ILK in HEL cells enhanced alpha(2)beta(1)-mediated adhesion whereas overexpression of kinase-dead ILK reduced adhesion, indicating a role for this kinase in the positive regulation of alpha(2)beta(1). Conclusions: Our findings that ILK regulates alpha(2)beta(1) in HEL cells, is activated in platelets and associates with beta(1)-integrins, raise the possibility that it may play a key role in adhesion events upon agonist stimulation of platelets.

Platelet endothelial cell adhesion molecule-1 signaling inhibits the activation of human platelets

Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is a 130-kd transmembrane glycoprotein and a member of the growing family of receptors with immunoreceptor tyrosine-based inhibitory motifs (ITIMs). PECAM-1 is expressed on platelets, certain T cells, monocytes, neutrophils, and vascular endothelial cells and is involved in a range of cellular processes, though the role of PECAM-1 in platelets is unclear. Cross-linking of PECAM-1 results in phosphorylation of the ITIM allowing the recruitment of signaling proteins that bind by way of Src-homology domain 2 interactions. Proteins that have been implicated in the negative regulation of cellular activation by ITIM-bearing receptors include the tyrosine phosphatases SHP-1 and SHP-2. Tyrosine phosphorylation of immunoreceptor tyrosine-based activatory motif (ITAM)-bearing receptors such as the collagen receptor GPVI-Fc receptor gamma-chain complex on platelets leads to activation. Increasing evidence suggests that ITIM- and ITAM-containing receptors may act antagonistically when expressed on the same cell. In this study it is demonstrated that cross-linking PECAM-1 inhibits the aggregation and secretion of platelets in response to collagen and the GPVI-selective agonist convulxin. In these experiments thrombin-mediated platelet aggregation and secretion were also reduced, albeit to a lesser degree than for collagen, suggesting that PECAM-1 function may not be restricted to the inhibition of ITAM-containing receptor pathways. PECAM-1 activation also inhibited platelet protein tyrosine phosphorylation stimulated by convulxin and thrombin; this was accompanied by inhibition of the mobilization of calcium from intracellular stores. These data suggest that PECAM-1 may play a role in the regulation of platelet function in vivo.

Search for narrow resonances and quantum black holes in inclusive and b-tagged dijet mass spectra from pp collisions at root s=7 TeV

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Search for W ' -> tb decays in the lepton plus jets final state in pp collisions at root s=8 TeV

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The cohomological invariant E'(G,W) and some properties

Let G be a group, W a nonempty G-set and M a Z2G-module. Consider the restriction map resG W : H1(G,M) → Pi wi∈E H1(Gwi,M), [f] → (resGG wi [f])i∈I , where E = {wi, i ∈ I} is a set of orbit representatives in W and Gwi = {g ∈ G | gwi = wi} is the G-stabilizer subgroup (or isotropy subgroup) of wi, for each wi ∈ E. In this work we analyze some results presented in Andrade et al [5] about splittings and duality of groups, using the point of view of Dicks and Dunwoody [10] and the invariant E'(G,W) := 1+dimkerresG W, defined when Gwi is a subgroup of infinite index in G for all wi in E, andM = Z2 (where dim = dimZ2). We observe that the theory of splittings of groups (amalgamated free product and HNN-groups) is inserted in the combinatory theory of groups which has many applications in graph theory (see, for example, Serre [12] and Dicks and Dunwoody [10]).

Tradução para o português brasileiro, adaptação transcultural e validação do 3D Assessment of Hand Function

Pós-graduação em Bases Gerais da Cirurgia - FMB

Perfil de aptidão cardiorrespiratória em corredores jovens pela cinética do VO2 em testes progressivos e retangulares

Pós-graduação em Desenvolvimento Humano e Tecnologias - IBRC

Scalar field description for parametric models of dark energy

We investigate theoretical and observational aspects of a time-dependent parameterization for the dark energy equation of state w(z), which is a well behaved function of the redshift z over the entire cosmological evolution, i.e., z is an element of [-1, infinity). By using a theoretical algorithm of constructing the quintes-sence potential directly from the w(z) function, we derive and discuss the general features of the resulting potential for the cases in which dark energy is separately conserved and when it is coupled to dark matter. Since the parameterization here discussed allows us to divide the parametric plane in defined regions associated to distinct classes of dark energy models, we use some of the most recent observations from type Ia supernovae, baryon acoustic oscillation peak and Cosmic Microwave Background shift parameter to check which class is observationally preferred. We show that the largest portion of the confidence contours lies into the region corresponding to a possible crossing of the so-called phantom divide line at some point of the cosmic evolution.

W Z plus missing-E-T signal from gaugino pair production at LHC7

LHC searches for supersymmetry currently focus on strongly produced sparticles, which are copiously produced if gluinos and squarks have masses of a few hundred GeV. However, in supersymmetric models with heavy scalars, as favored by the decoupling solution to the SUSY flavor and CP problems, and m((g) over tilde) greater than or similar to 500 GeV as indicated by recent LHC results, chargino-neutralino ((W) over tilde (+/-)(1)(Z) over tilde (2)) production is the dominant cross section for m((W) over tilde1) similar to m((Z) over tilde2) < m(<(g)over tilde>)/3 at LHC with root s = 7 TeV (LHC7). Furthermore, if m((Z) over tilde1) + (m (Z) over tilde) less than or similar to m((Z) over tilde2) less than or similar to m((Z) over tilde1) + m(h), then (Z) over tilde (2) dominantly decays via (Z) over tilde (2) -> (Z) over tilde (1)Z, while (W) over tilde (1) decays via (W) over tilde (1) -> (Z) over tilde W-1. We investigate the LHC7 reach in the W Z + (sic)T channel (for both leptonic and hadronic decays of the W boson) in models with and without the assumption of gaugino mass universality. In the case of the mSUGRA/CMSSM model with heavy squark masses, the LHC7 discovery reach in the W Z+ (sic)T channel becomes competetive with the reach in the canonical (sic)T + jets channel for integrated luminosities similar to 30 fb(-1). We also present the LHC7 reach for a simplified model with arbitrary m((Z) over tilde1) and m((W) over tilde1) similar to m((Z) over tilde2). Here, we find a reach of up to m((W) over tilde1) similar to 200 (250) GeV for 10 (30) fb(-1).

CD69 modulates sphingosine-1-phosphate-induced migration of skin dendritic cells

In this study, we have investigated the role of CD69, an early inducible leukocyte activation receptor, in murine dendritic cell (DC) differentiation, maturation, and migration. Skin DCs and DC subsets present in mouse lymphoid organs express CD69 in response to maturation stimuli. Using a contact sensitization model, we show that skin DCs migrated more efficiently to draining lymph nodes (LNs) in the absence of CD69. This was confirmed by subcutaneous transfer of CD69-/- DCs, which presented an increased migration to peripheral LNs. Two-photon microscopy analysis showed that once DCs reached the LNs, CD69 deficiency did not alter DC interstitial motility in the LNs. Chemotaxis to sphingosine-1-phosphate (S1P) was enhanced in CD69-/- DCs compared with wild-type DCs. Accordingly, we detected a higher expression of S1P receptor type-1 (S1P(1)) by CD69-/- DCs, whereas S1P(3) expression levels were similar in wild-type and CD69-/- DCs. Moreover, in vivo treatment with S1P analogs SEW2871 and FTY720 during skin sensitization reduced skin DC migration to peripheral LNs. These results suggest that CD69 regulates S1P-induced skin DC migration by modulating S1P(1) function. Together, our findings increase our knowledge on DC trafficking patterns in the skin, enabling the development of new directed therapies using DCs for antigen (Ag) delivery.

Na 1 Nachlass Max Horkheimer, 643 - Korrekturen zur Neupublikation der Aufsätze Max Horkheimers aus der Zeitschrift für Sozialforschung und Vorwort zur Neupublikation 1968 (p. IX 23.1-20-IX 24.1-9)

1. Friedrich Pollock: Verzeichnis der Änderungen in den Essays nach der Bearbeitung durch Alfred Schmidt, 4 Blatt; 2. Derselbe: Eigenhändige Korrekturnotiz zum Manuskript Schmidt, 1 Blatt; 3. Derselbe: Eigenhändige Notiz über Verteilung von Korrekturen, 1 Blatt; 4. Derselbe: Ergänzungen zu den Korrekturen vom 15. und 20.03.1968. a) 8 Blatt b) 8 Blatt; 5. Korrekturen, 2 Blatt; 6. Friedrich Pollock [?]: Verzeichnis der Essays von Max Horkheimer und der Korrekturen. a) 8 Blatt b) Teilstücke, 2 Blatt; 7. Derselbe [?]: Verzeichnis der Korrekturen, 3 Blatt; 8. Derselbe: Eigenhändige Gesprächsnotiz, 1 Blatt; 9. Derselbe: Eigenhändige Gesprächsnotiz, 1 Blatt; 10. Derselbe: "Sprachregeln", eigenhändige Notiz, 1 Blatt; 11. Derselbe: "Bedenkliche Stellen", eigenhändiges Verzeichnis, 1 Blatt; 12. Derselbe: "Vorschlag für den Inhalt von Max Horkheimers Essays I", eigenhändiges Verzeichnis, 1 Blatt; 13. Derselbe: Eigenhändige Gesprächsnotiz Friedrich Pollocks - Max Horkheimers über Pro und Contra, Neuveröffentlichung, 1 Blatt; 14. S. Fischer Verlaf: Schätzung des Umfangs der Essys, 1 Blatt; 15. Friedrich Pollock: Verzeichnis der Essays von Max Horkheimer, 3 Blatt; 16. Handschriftliches Verzeichnis der Aufsätze, Reden und Schriften Max Horkheimers, 4 Blatt; 17. "Max Horkheimer Essays" Verzeichnis, 1 Blatt; 18. Schönbach, Peter: 1 Brief mit Unterschrift an Max Horkheimer Frankfurt, 11.09.1964, 7 Blatt; 19. Liste der Anmerkungen Frau Dr. Adornos zu den Aufsätzen Prof. Horkheimers in der Zeitschrift für Sozialforschung, 2 Blatt; 20. Adorno, Gretel: 4 Briefe mit Unterschrift an Max Horkheimer, Korrekturvorschläge Frankfurt am Main 1962-1963, 5 Blatt; Vowort zur Neupublikation 1968 der Aufsätze aus der Zeitschrift für Sozialforschung (GS 3, S. 14-19); 1964-1968; Veröffentlicht in: Max Horkheimer "Kritische Theorie", Frankfurt am Main 1968, Seite IX- IXV; 1. Entwürfe Juli- September 1968; 2. Kalb, Peter E.: 1 Brief an Max Horkheimer und Beilage, Frankfurt am Main, 02.08.1968, 2 Blatt; 3. Klappentext der Buchausgabe, Korrekturfahne mit handschriftlichen Korrekturen; 4. Adorno, Theodor W.: 1 Brief mit Unterschrift an Max Horkheimer, Frankfurt, 17.07.1968, 1 Blatt; 5. Horkheimer, Max: "Der neuste Angriff auf die Metaphysik". Sonderdruck der Zeitschrift für Sozialforschung VI, 1937, mit eigenhändigen Korrekturen; 6. Derselbe: "Autoritärer Staat" Teilstücke aus der Gedenkschrift für Walter Benjamin, 1942. Als Typoskript vervielfältigt, 21 Blatt; 7. "Zitate aus 'Autoritärer Staat' heute", 4 Blatt; 8. Schmidt, Alfred: "Zur Idee der kritischen Theorie" = Nachwort zur Buchausgabe. Typoskript mit handschriftlichen Korrekturen von Max Horkheimer, 38 Blatt; 9. N.N.: Ergänzungsvorschläge zu dem Nachwort von Dr. Schmidt, 3 Blatt;

Na 1 Nachlass Max Horkheimer, 712 - Entwürfe für ein mögliches Forschungsprojekt über Liberalismus des 19. Jahrhunderts; "Untersuchung des sozialen Klimas in Stadt- und Landkreisen Hessens"; "Betriebsklime. Eine industrie-soziologische Untersuchung aus dem Ruhrgebiet"; Sardemann, Karl: Interview mit einem Juden über sein Leben 1933 bis circa 1953; "Altersbild und Altersvorsorge der Arbeiter und Angestellte" (p. IX 187 - IX 191.1-3)

Entwürfe für ein Mögliches Forschungsprojekt über Liberlaismus des 19. Jahrhunderts; 1953; 1. Typoskript mit eigenhändigen Korrekturen von Theodor W. Adorno; 7 Blatt; 2. Typoskript mit eigenhändigen Korrekturen von Theodor W. Adorno; 3 Blatt; "Untersuchungn des sozialen Klimas in Stadt- und Landkreisen Hessens"; Entwürfe für ein "Handbuch der Methoden zur Messung des sozialen Klimas", nicht veröffentlicht; 1953; 1. Pollock, Friedrich: "Einführung" zum "Handbuch" a) Typoskript mit eigenhändigen Korrekturen; 8 Blatt; b) Typoskript mit eigenhändigen Korrekturen, 5 Blatt; c) Manuskript, "Vorwort"; 5 Blatt; d) Baumert, Gerhard: Entwurf des "Vorworts"; Typoskript mit handschriftlichen Korrekturen, 2 Blatt; e) Baumert, Gerhard: "Notizen zu Vorwort und Einführung des Handbuchs"; Typoskript, 1 Blatt; 2. Inhaltsverzeichnis zum "Handbuch"; 1 Blatt; 3. Pollock, Friedrich: eigenhändige Notizen zur Einführung; 7 Blatt; 4. Pollock, Friedrich: 1 eigenhändiger Brief mit Unterschrift an Theodor W. Adorno, Santa Monica, 21.09.1953; 1 Blatt; "Betriebsklime. Eine industrie-soziologische Untersuchung aus dem Ruhrgebiet" 1954-1956 veröffentlicht als Band 3 der Frankfurter Beiträge zur Soziologie, Frankfurt 1955; 1. Druckfahnen, mit handschriftlichen Korrekturen; 38 Blatt; 2. Korrektur-Notizen zu den Druckfahnen; 4 Blatt; 3. Adorno, Theodor W. [Mitarbeit]: "Grundreiz [Betriebsumfrage)" a) Typoskript mit handschriftlichen Korrekturen, 4 Blatt; b) Typoskript, 5 Blatt; 4. Adorno, Theodor W.: 1 Brief an Max Horkheimer, ohne Ort, 30.06.1954; 1 Blatt; 5. Fragebogen- Entwurf; Typsokript, 2 Blatt; 6. Fragebogen- Entwurf; Typoskript, 2 Blatt; 7. Adorno, Theodor W.: 1 Brief an Max Horkheimer, ohne Ort, ohne Datum [1954]; 1 Blatt; 8. Dirks, Walter: "Notiz über meine Reise nach Köln, Düsseldorf und Essen", 22.02.1955. Typoskript, 2 Blatt; 9. Becker, Hellmut: "Aktennotiz, Betreff: Ersetzung der Pressekonferenz über die Mannesmann-Studie durch in den nächsten Wochen und Monaten aufeinanderfolgende Artikel qualifizierter Korrespondenten", 15.02.1955. Typoskript, 1 Blatt; 10. Adorno, Theodor W.: 1 Brief an Hermann Winkhaus, Mannesmann AG, ohne Ort, 05.03.1956; 2 Blatt; 11. Winkhaus, Hermann, Mannesmann AG: 1 Breifabschrift an das Institut für Sozialforschung, Düsseldorf, 18.07.1955; 12. Presseveröffentlichungen zur Betriebsklima-Untersuchung; 1 Ordner, 13 Blatt; 13. Zeitschriftenartikel und Abschriften von Artikeln zur Betriebsklima- Untersuchung; mit: Becker, Egon: 2 Briefe mit Unterschrift an Max Horkheimer, Frankfurt, 1955; 1 Brief von Max Horkheimer, Zürich, 25.10.1955; Sardemann, Karl: Interview mit einem Juden über sein Leben 1933 bis circa 1953, insbesondere seine Erlebnisse im Konzentrationslager; 1955; 1. Interview; Typsokript, 38 Blatt; 2. Sardemann, Karl: 1 Brief mit Unterschrift an Max Horkheimer, ohne Ort, 15.12.1955; 1 Blatt; "Altersbild und Altersvorsorge der Arbeiter und Angestellten" veröffentlicht als Sonderheft 1 der Frankfurter Beiträge zur Soziologie, Frankfurt 1958.; Zwischenbericht zum Projekt und Material, 1955; 1. Becker, Egon, u.a.: "Zwischenbericht" 10.03.1953; Typoskript, 9 Blatt; mit einem Brief mit Unterschrift von Egon Becker an Max Horkheimer, ohne Ort, 11.03.1955; 1 Blatt; 2. Fragebogen; als Typoskript vervielfältigt, 16 Blatt; 3. Fragebogen; als Typoskript vervielfältigt, 11 Blatt;

Characterization of the NOP -1 opsin photoreceptor in the filamentous fungus Neurospora crassa

Light absorption is an important process for energy production and sensory perception in many organisms. In the filamentous fungus, Neurospora crassa, blue-light is an important regulator of both asexual and sexual development, but the identity of the blue-light receptor is unknown. The work presented in this dissertation initiated the characterization of the putative N. crassa opsin photoreceptor, NOP-1. Opsins were thought to exist only in the archaea and mammals until the discovery of nop-1. All opsins have the same conserved structure of seven transmembrane helical domains with a lysine residue in the seventh helix specific for forming a Schiff-base linkage with retinal. The predicted NOP-1 protein sequence is equally similar to archaeal rhodopsins and a newly identified fungal opsin-related protein group (ORPs). ORPs maintain the seven transmembrane helical structure of opsins, but lack the conserved lysine residue for binding retinal. An ORP gene, orp-1 was identified in N. crassa and this work includes the cloning and sequence analysis of this gene. Characterization of NOP-1 function in N. crassa development began with the construction of a Δnop-1 deletion mutant. Extensive phenotypic analysis of Δnop-1 mutants revealed only subtle defects during development primarily under environmental conditions that induce a stress response. NOP-1 was overexpressed in the heterologous system Pichia pastoris, and it was demonstrated that NOP-1 protein bound all-trans retinal to form a green-light absorbing pigment (λmax = 534 nm) with a photochemical reaction cycle similar to archaeal sensory rhodopsins. nop-1 gene expression was monitored during N. crassa development. nop-1 transcript is highly expressed during asexual sporulation (conidiation) and transcript levels are abundant in the later stages of conidial development. nop-1 expression is not regulated by blue-light or elevated temperatures. Potential functions for NOP-1 were discovered through the transcriptional analysis of conidiation-associated genes in Δnop-1 mutants. NOP-1 exhibits antagonistic transcriptional regulation of conidiation-associated genes late in conidial development, by enhancing the carotenogenic gene, al-2 and repressing the conidiation-specific genes, con-10 and con-13. ^