Microtubule Associated Protein 1b (MAP1B) Is a Marker of the Microtubular Cytoskeleton in Podocytes but Is Not Essential for the Function of the Kidney Filtration Barrier in Mice.

Podocytes are essential for the function of the kidney glomerular filter. A highly differentiated cytoskeleton is requisite for their integrity. Although much knowledge has been gained on the organization of cortical actin networks in podocyte's foot processes, less is known about the molecular organization of the microtubular cytoskeleton in primary processes and the cell body. To gain an insight into the organization of the microtubular cytoskeleton of the podocyte, we systematically analyzed the expression of microtubule associated proteins (Maps), a family of microtubules interacting proteins with known functions as regulator, scaffold and guidance proteins. We identified microtubule associated protein 1b (MAP1B) to be specifically enriched in podocytes in human and rodent kidney. Using immunogold labeling in electron microscopy, we were able to demonstrate an enrichment of MAP1B in primary processes. A similar association of MAP1B with the microtubule cytoskeleton was detected in cultured podocytes. Subcellular distribution of MAP1B HC and LC1 was analyzed using a double fluorescent reporter MAP1B fusion protein. Subsequently we analyzed mice constitutively depleted of MAP1B. Interestingly, MAP1B KO was not associated with any functional or structural alterations pointing towards a redundancy of MAP proteins in podocytes. In summary, we established MAP1B as a specific marker protein of the podocyte microtubular cytoskeleton.

La teocrazia : crisi e trasformazione di un modello politico nell'Europa del XVIII secolo

Ma recherche porte sur la crise et les diverses transformations subies par la théocratie en tant que modèle politique en Europe dans la première moitié du XVIIIe siècle. Mon attention se concentre sur la remise en question de la valeur normative, à savoir sacrée, que ce modèle avait acquis au cours du XVIe et du XVIIe siècle, et sur sa transformation en un modèle universel et «négatif» au cours de la première partie du XVIIIe siècle. L'analyse du modèle théocratique n'a pas fait, jusqu'à présent, l'objet de nombreuses études de la part des historiens de la pensée politique moderne. Il est resté au centre de la réflexion d'autres domaines d'études. Seulement, depuis les années 1990, certaines études historiographiques ont montré l'existence, au cours des XVIe et XVIIe siècles, d'un grand intérêt autour de la normatività du modèle théocratique. L'analyse portait sur une série de traités relatifs à l'utilisation politique de la théocratie juive, c'est-à-dire le gouvernement de l'ancien Israël ou respublica Hebraeorum, telle qu'elle était décrite dans le texte sacré. La limite de ces études est cependant d'avoir analysé seulement « l'âge d'or » de cette littérature, à savoir les XVIe et XVIIe siècles. Ils ont identifié dans le Tractatus Theo logico -Poäücus de Spinoza le déclin de l'intérêt européen pour ce modèle politique. Ma recherche vise à mettre en évidence l'évolution de la façon de considérer la théocratie en tant que. modèle politique en Europe au XVIIIe siècle, dans le but de démontrer que le Tractatus Theologico- Poäticus n'a pas signé son déclin définitif en Europe en soulignant comment le débat politique de cette époque a été caractérisé par un fort intérêt politique pour la théocratie. En particulier mon analyse porte sur trois moments différents. La première partie de mon étude concerne la remise en question de la valeur positive et normative du modèle, théocratique juif à travers son historicisation. A cet effet, je me concentre sur trois figures emblématiques, à savoir Giambattista Vico, John Toland et Jacques Basnage en démontrant comment ce trois auteurs, chacun de leur côté, ont contribué au renversement de l'image classique de la respublica Hebraeorum. Dans un deuxième temps mon étude analyse la phase de radicalisation de ces changements au milieu du XVIIIe siècle et, en particulier, dans les ouvrages de Nicolas Antoine Boulanger, les Recherches sur l'origine du despotisme oriental (1761) et l'Antiquité dévoilée par ses usags (1765). Boulanger fut le premier à fournir une explication de la naissance des différentes formes de gouvernements politiques et des différentes religions en se référant à de grandes catastrophes géologiques, notamment le Déluge. La théocratie s'explique ainsi par un millénarisme aigu suite à l'expérience catastrophique de l'inondation. Le modèle décrite par Boulanger se distingue clairement de celui de la littérature, précédente. La théocratie subit un déplacement sémantique de modèle politique associé à l'histoire juive avec une connotation «positive» à un modèle universel à connotation totalement la «négative». Dans cette transition, qui s'est produite de manière progressive entre le XVIIe et le XVIIIe siècle, le modèle biblique perd sa principale caractéristique, à savoir sa sacralité. La troisième et dernière partie de cette étude est une brève description de l'héritage du modèle théocratique dans XEncyclopédie ou Dictionnaire raisonné des sciences, des arts et des métiers. En particulier, l'analyse se concentre sur l'article «Théocratie» et l'article «OEconomie politique». Ma recherche vise à montrer les éléments en commun de ces deux articles avec les ouvrages de Boulanger. Ces deux contributions sont présentées dans mon travail comme un moyen de transmission de la pensée de Boulanger dans l'Encyclopedie.

Buried in the Middle but Guilty: Intronic Mutations in the TCIRG1 Gene Cause Human Autosomal Recessive Osteopetrosis.

Autosomal recessive osteopetrosis (ARO) is a rare genetic bone disease with genotypic and phenotypic heterogeneity, sometimes translating into delayed diagnosis and treatment. In particular, cases of intermediate severity often constitute a diagnostic challenge and represent good candidates for exome sequencing. Here, we describe the tortuous path to identification of the molecular defect in two siblings, in which osteopetrosis diagnosed in early childhood followed a milder course, allowing them to reach the adult age in relatively good conditions with no specific therapy. No clearly pathogenic mutation was identified either with standard amplification and resequencing protocols or with exome sequencing analysis. While evaluating the possible impact of a 3'UTR variant on the TCIRG1 expression, we found a novel single nucleotide change buried in the middle of intron 15 of the TCIRG1 gene, about 150 nucleotides away from the closest canonical splice site. By sequencing a number of independent cDNA clones covering exons 14 to 17, we demonstrated that this mutation reduced splicing efficiency but did not completely abrogate the production of the normal transcript. Prompted by this finding, we sequenced the same genomic region in 33 patients from our unresolved ARO cohort and found three additional novel single nucleotide changes in a similar location and with a predicted disruptive effect on splicing, further confirmed in one of them at the transcript level. Overall, we identified an intronic region in TCIRG1 that seems to be particularly prone to splicing mutations, allowing the production of a small amount of protein sufficient to reduce the severity of the phenotype usually associated with TCIRG1 defects. On this basis, we would recommend including TCIRG1 not only in the molecular work-up of severe infantile osteopetrosis but also in intermediate cases and carefully evaluating the possible effects of intronic changes. © 2015 American Society for Bone and Mineral Research.

Protease inhibitors to treat hepatitis C in the Swiss HIV Cohort Study: high efficacy but low treatment uptake.

OBJECTIVES: Direct-acting antiviral agents (DAAs) have become the standard of care for the treatment of chronic hepatitis C virus (HCV) infection. We aimed to assess treatment uptake and efficacy in routine clinical settings among HIV/HCV coinfected patients after the introduction of the first generation DAAs. METHODS: Data on all Swiss HIV Cohort Study (SHCS) participants starting HCV protease inhibitor (PI) treatment between September 2011 and August 2013 were collected prospectively. The uptake and efficacy of HCV therapy were compared with those in the time period before the availability of PIs. RESULTS: Upon approval of PI treatment in Switzerland in September 2011, 516 SHCS participants had chronic HCV genotype 1 infection. Of these, 57 (11%) started HCV treatment during the following 2 years with either telaprevir, faldaprevir or boceprevir. Twenty-seven (47%) patients were treatment-naïve, nine (16%) were patients with relapse and 21 (37%) were partial or null responders. Twenty-nine (57%) had advanced fibrosis and 15 (29%) had cirrhosis. End-of-treatment virological response was 84% in treatment-naïve patients, 88% in patients with relapse and 62% in previous nonresponders. Sustained virological response was 78%, 86% and 40% in treatment-naïve patients, patients with relapse and nonresponders, respectively. Treatment uptake was similar before (3.8 per 100 patient-years) and after (6.1 per 100 patient-years) the introduction of PIs, while treatment efficacy increased considerably after the introduction of PIs. CONCLUSIONS: The introduction of PI-based HCV treatment in HIV/HCV-coinfected patients improved virological response rates, while treatment uptake remained low. Therefore, the introduction of PIs into the clinical routine was beneficial at the individual level, but had only a modest effect on the burden of HCV infection at the population level.

Evidence for daily and weekly rhythmicity but not lunar or seasonal rhythmicity of physical activity in a large cohort of individuals from five different countries.

Background Biological rhythmicity has been extensively studied in animals for many decades. Although temporal patterns of physical activity have been identified in humans, no large-scale, multi-national study has been published, and no comparison has been attempted of the ubiquity of activity rhythms at different time scales (such as daily, weekly, monthly, and annual). Methods Using individually worn actigraphy devices, physical activity of 2,328 individuals from five different countries (adults of African descent from Ghana, South Africa, Jamaica, Seychelles, and the United States) was measured for seven consecutive days at different times of the year. Results Analysis for rhythmic patterns identified daily rhythmicity of physical activity in all five of the represented nationalities. Weekly rhythmicity was found in some, but not all, of the nationalities. No significant evidence of lunar rhythmicity or seasonal rhythmicity was found in any of the groups. Conclusions These findings extend previous small-scale observations of daily rhythmicity to a large cohort of individuals from around the world. The findings also confirm the existence of modest weekly rhythmicity but not lunar or seasonal rhythmicity in human activity. These differences in rhythm strength have implications for the management of health hazards of rhythm misalignment. Key Messages Analysis of the pattern of physical activity of 2,328 individuals from five countries revealed strong daily rhythmicity in all five countries, moderate weekly rhythmicity in some countries, and no lunar rhythmicity or seasonal rhythmicity in any of the countries.

Dpp spreading is required for medial but not for lateral wing disc growth.

Drosophila Decapentaplegic (Dpp) has served as a paradigm to study morphogen-dependent growth control. However, the role of a Dpp gradient in tissue growth remains highly controversial. Two fundamentally different models have been proposed: the 'temporal rule' model suggests that all cells of the wing imaginal disc divide upon a 50% increase in Dpp signalling, whereas the 'growth equalization model' suggests that Dpp is only essential for proliferation control of the central cells. Here, to discriminate between these two models, we generated and used morphotrap, a membrane-tethered anti-green fluorescent protein (GFP) nanobody, which enables immobilization of enhanced (e)GFP::Dpp on the cell surface, thereby abolishing Dpp gradient formation. We find that in the absence of Dpp spreading, wing disc patterning is lost; however, lateral cells still divide at normal rates. These data are consistent with the growth equalization model, but do not fit a global temporal rule model in the wing imaginal disc.

NLRP12 is a neutrophil-specific, negative regulator of in vitro cell migration but does not modulate LPS- or infection-induced NF-κB or ERK signalling.

NOD-like receptors (NLR) are a family of cytosolic pattern recognition receptors that include many key drivers of innate immune responses. NLRP12 is an emerging member of the NLR family that is closely related to the well-known inflammasome scaffold, NLRP3. Since its discovery, various functions have been proposed for NLRP12, including the positive regulation of dendritic cell (DC) and neutrophil migration and the inhibition of NF-κB and ERK signalling in DC and macrophages. We show here that NLRP12 is poorly expressed in murine macrophages and DC, but is strongly expressed in neutrophils. Using myeloid cells from WT and Nlrp12(-/)(-) mice, we show that, contrary to previous reports, NLRP12 does not suppress LPS- or infection-induced NF-κB or ERK activation in myeloid cells, and is not required for DC migration in vitro. Surprisingly, we found that Nlrp12 deficiency caused increased rather than decreased neutrophil migration towards the chemokine CXCL1 and the neutrophil parasite Leishmania major, revealing NLRP12 as a negative regulator of directed neutrophil migration under these conditions.

Gli altri noi : Rom e residenti nella Svizzera italiana : etnografia e mediazione

Cette recherche porte sur les roms (vlach) qui séjournent périodiquement en Suisse italienne, en camping-cars ou en caravanes, pratiquant un semi- nomadisme lié à des motivations principalement économiques, mais également familiales et religieuses. Elle poursuit simultanément deux objectifs. Le premier est lié à un souci de médiation culturelle, qui rejoint les réflexions sur la façon de gérer au mieux les conflits entre les diverses composantes du vivre ensemble. Le second vise à approfondir la connaissance des populations roms concernées (lovara, éurara ou kalderasa, de nationalités italienne, française et espagnole). Les deux objectifs sont évidemment reliés, car comprendre les dynamiques conflictuelles exige la connaissance approfondie des acteurs en interaction. Les relations entre résidents tessinois et roms ont évolué dans le temps, avec des oscillations entre le pôle du refus et celui de l'attraction, entre rejet et reconnaissance, méfiance et confiance. Ces variations historiques renvoient à un fondement commun, à une dynamique des oppositions réciproques. L'analyse anthropologique a fait apparaître le rôle ambivalent du conflit dans la production et la reproduction de la relation d'altérité. L'exclusion des roms fournit à certaines tendances politiques locales l'occasion d'apparaître comme les protecteurs des résidents contre les étrangers ; inversement, l'opposition des roms aux gaiés leur permet de resserrer leur communauté et de prévenir l'assimilation. La médiation culturelle peut espérer gérer cette dynamique relationnelle, certainement pas la faire disparaître. Le modèle intégratif traditionnel de l'État- nation se révèle trop étroit pour accueillir la culture rom. Casser la résistance des roms envers le travail salarié et envers la scolarisation obligatoire, entraver leur semi-nomadisme (qui incite aussi leurs rencontres familiales et religieuses), refuser de reconnaître leur organe de justice interne et leur langue, tout cela ne ferait que concourir à leur disparition culturelle. Les roms sont conscients de ces possibilités, raison pour laquelle ils maintiennent une ambivalence relationnelle avec les ga£és, quitte à apparaître comme des « tsiganes indésirables ». Abstract : This research focuses on the Roma people (Vlach) who periodically stay in Italian Switzerland in caravans or camper vans, practising a semi-nomadic lifestyle - a choice based mainly on economic imperatives but also on familial and religious grounds. This study will address two objectives at the same time. The first involves cultural mediation, and considers the best way to manage the existing conflicts between the cultural components of the social environment. The second aims to increase knowledge of the Roma populations under examination - the Italian, French and Spanish Lovara, Éurara or Kalderasa. The two objectives are clearly linked, as a deeper understanding of the people involved is vital if we are to increase understanding of the dynamics of confrontation. The relationships between the residents of the Ticino area and the Roma people have evolved over time, oscillating in varying degrees between rejection and attraction, confrontation and recognition, suspicion and trust. These variations spring from a common base, a dynamic shaped by mutual opposition. The anthropological analysis highlights the ambivalent role of conflict in the production and reproduction of relations of otherness. Excluding the Roma has given some local politicians the opportunity to position themselves as protectors of residents against foreigners. Inversely, the Roma people's opposition to the Gazé strengthens links within their community and prevents assimilation. Cultural mediation may aspire to manage this relational dynamic, but not abolish it. The traditional integrative model of the nation-state has shown itself to be too restrictive to accommodate the Romani culture. Rejecting the Roma's resistance to salaried work and obligatory schooling, obstructing their semi-nomadic lifestyle (one that also involves family and religious assemblies) and refusing to recognise their traditional courts and mother tongue, all that can only contribute to the dilution of their cultural heritage. The Roma are fully aware of these possibilities, which is why they retain a relational ambivalence in their dealings with the Gazé, at the cost of appearing to be "undesirable gypsies".

Methotrexate Is Not Superior to Placebo for Inducing Steroid-Free Remission, but Induces Steroid-Free Clinical Remission in a Larger Proportion of Patients With Ulcerative Colitis.

BACKGROUND & AIMS: Parenteral methotrexate is an effective treatment for patients with Crohn's disease, but has never been adequately evaluated in patients with ulcerative colitis (UC). We conducted a randomized controlled trial to determine its safety and efficacy in patients with steroid-dependent UC. METHODS: We performed a double-blind, placebo-controlled trial to evaluate the efficacy of parenteral methotrexate (25 mg/wk) in 111 patients with corticosteroid-dependent UC at 26 medical centers in Europe from 2007 through 2013. Patients were given prednisone (10 to 40 mg/d) when the study began and were randomly assigned to groups (1:1) given placebo or methotrexate (intramuscularly or subcutaneously, 25 mg weekly) for 24 weeks. The primary end point was steroid-free remission (defined as a Mayo score ≤2 with no item >1 and complete withdrawal of steroids) at week 16. Secondary endpoints included clinical remission (defined as a Mayo clinical subscore ≤2 with no item >1) and endoscopic healing without steroids at weeks 16 and/or 24, remission without steroids at week 24, and remission at both weeks 16 and 24. RESULTS: Steroid-free remission at week 16 was achieved by 19 of 60 patients given methotrexate (31.7%) and 10 of 51 patients given placebo (19.6%)-a difference of 12.1% (95% confidence interval [CI]: -4.0% to 28.1%; P = .15). The proportion of patients in steroid-free clinical remission at week 16 was 41.7% in the methotrexate group and 23.5% in the placebo group, for a difference of 18.1% (95% CI: 1.1% to 35.2%; P = .04). The proportions of patients with steroid-free endoscopic healing at week 16 were 35% in the methotrexate group and 25.5% in the placebo group-a difference of 9.5% (95% CI: -7.5% to 26.5%; P = .28). No differences were observed in other secondary end points. More patients receiving placebo discontinued the study because of adverse events (47.1%), mostly caused by UC, than patients receiving methotrexate (26.7%; P = .03). A higher proportion of patients in the methotrexate group had nausea and vomiting (21.7%) than in the placebo group (3.9%; P = .006). CONCLUSIONS: In a randomized controlled trial, parenteral methotrexate was not superior to placebo for induction of steroid-free remission in patients with UC. However, methotrexate induced clinical remission without steroids in a significantly larger percentage of patients, resulting in fewer withdrawals from therapy due to active UC. ClinicalTrials.gov ID NCT00498589.

Galactofuranose in Mycoplasma mycoides is important for membrane integrity and conceals adhesins but does not contribute to serum resistance.

Mycoplasma mycoides subsp. capri (Mmc) and subsp. mycoides (Mmm) are important ruminant pathogens worldwide causing diseases such as pleuropneumonia, mastitis and septicaemia. They express galactofuranose residues on their surface, but their role in pathogenesis has not yet been determined. The M. mycoides genomes contain up to several copies of the glf gene, which encodes an enzyme catalysing the last step in the synthesis of galactofuranose. We generated a deletion of the glf gene in a strain of Mmc using genome transplantation and tandem repeat endonuclease coupled cleavage (TREC) with yeast as an intermediary host for the genome editing. As expected, the resulting YCp1.1-Δglf strain did not produce the galactofuranose-containing glycans as shown by immunoblots and immuno-electronmicroscopy employing a galactofuranose specific monoclonal antibody. The mutant lacking galactofuranose exhibited a decreased growth rate and a significantly enhanced adhesion to small ruminant cells. The mutant was also 'leaking' as revealed by a β-galactosidase-based assay employing a membrane impermeable substrate. These findings indicate that galactofuranose-containing polysaccharides conceal adhesins and are important for membrane integrity. Unexpectedly, the mutant strain showed increased serum resistance.

High Altitude Increases Alteration in Maximal Torque but Not in Rapid Torque Development in Knee Extensors after Repeated Treadmill Sprinting.

We assessed knee extensor neuromuscular adjustments following repeated treadmill sprints in different normobaric hypoxia conditions, with special reference to rapid muscle torque production capacity. Thirteen team- and racquet-sport athletes undertook 8 × 5-s "all-out" sprints (passive recovery = 25 s) on a non-motorized treadmill in normoxia (NM; FiO2 = 20.9%), at low (LA; FiO2 = 16.8%) and high (HA; FiO2 = 13.3%) normobaric hypoxia (simulated altitudes of ~1800 m and ~3600 m, respectively). Explosive (~1 s; "fast" instruction) and maximal (~5 s; "hard" instruction) voluntary isometric contractions (MVC) of the knee extensors (KE), with concurrent electromyographic (EMG) activity recordings of the vastus lateralis (VL) and rectus femoris (RF) muscles, were performed before and 1-min post-exercise. Rate of torque development (RTD) and EMG (i.e., Root Mean Square or RMS) rise from 0 to 30, -50, -100, and -200 ms were recorded, and were also normalized to maximal torque and EMG values, respectively. Distance covered during the first 5-s sprint was similar (P > 0.05) in all conditions. A larger (P < 0.05) sprint decrement score and a shorter (P < 0.05) cumulated distance covered over the eight sprints occurred in HA (-8 ± 4% and 178 ± 11 m) but not in LA (-7 ± 3% and 181 ± 10 m) compared to NM (-5 ± 2% and 183 ± 9 m). Compared to NM (-9 ± 7%), a larger (P < 0.05) reduction in MVC torque occurred post-exercise in HA (-14 ± 9%) but not in LA (-12 ± 7%), with no difference between NM and LA (P > 0.05). Irrespectively of condition (P > 0.05), peak RTD (-6 ± 11%; P < 0.05), and normalized peak RMS activity for VL (-8 ± 11%; P = 0.07) and RF (-14 ± 11%; P < 0.01) muscles were reduced post-exercise, whereas reductions (P < 0.05) in absolute RTD occurred within the 0-100 (-8 ± 9%) and 0-200 ms (-10 ± 8%) epochs after contraction onset. After normalization to MVC torque, there was no difference in RTD values. Additionally, the EMG rise for VL muscle was similar (P > 0.05), whereas it increased (P < 0.05) for RF muscle during all epochs post-exercise, independently of the conditions. In summary, alteration in repeated-sprint ability and post-exercise MVC decrease were greater at high altitude than in normoxia or at low altitude. However, the post-exercise alterations in RTD were similar between normoxia and low-to-high hypoxia.