Targeted NGS gene panel identifies mutations in RSPH1 causing primary ciliary dyskinesia and a common mechanism for ciliary central pair agenesis due to radial spoke defects.

Primary ciliary dyskinesia (PCD) is an inherited chronic respiratory obstructive disease with randomized body laterality and infertility, resulting from cilia and sperm dysmotility. PCD is characterized by clinical variability and extensive genetic heterogeneity, associated with different cilia ultrastructural defects and mutations identified in >20 genes. Next generation sequencing (NGS) technologies therefore present a promising approach for genetic diagnosis which is not yet in routine use. We developed a targeted panel-based NGS pipeline to identify mutations by sequencing of selected candidate genes in 70 genetically undefined PCD patients. This detected loss-of-function RSPH1 mutations in four individuals with isolated central pair (CP) agenesis and normal body laterality, from two unrelated families. Ultrastructural analysis in RSPH1-mutated cilia revealed transposition of peripheral outer microtubules into the 'empty' CP space, accompanied by a distinctive intermittent loss of the central pair microtubules. We find that mutations in RSPH1, RSPH4A and RSPH9, which all encode homologs of components of the 'head' structure of ciliary radial spoke complexes identified in Chlamydomonas, cause clinical phenotypes that appear to be indistinguishable except at the gene level. By high-resolution immunofluorescence we identified a loss of RSPH4A and RSPH9 along with RSPH1 from RSPH1-mutated cilia, suggesting RSPH1 mutations may result in loss of the entire spoke head structure. CP loss is seen in up to 28% of PCD cases, in whom laterality determination specified by CP-less embryonic node cilia remains undisturbed. We propose this defect could arise from instability or agenesis of the ciliary central microtubules due to loss of their normal radial spoke head tethering

Implementation of optimized supportive care and hospital needs along the management of patients with advanced lung cancer

Background: Supportive care in cancer (SCC) have been recommended to be integrated in the management of patients with lung cancer all along the course of the disease. We took advantage of a pilot program of early implementation of optimized SCC, to report the feasibility such program in patients with advanced lung cancer, and correlate patient characteristics and outcomes with the actual use of optimized SCC.Methods: This study is a retrospective analysis of all consecutive patients with lung cancer treated at our center between 2012 and 2016. Optimized SCC included the intervention of a nurse for the home-hospital network coordination, as well as socio-aesthetics, psychomotricity, art-therapy, adapted physical activity, and also establishment of at-home hospitalization.Results: 309 patients were included. Median overall survival was 11.2 months. Unplanned hospitalizations occurred for 276 (89%) patients. The median duration of hospital stay was 19 days. Unplanned hospitalizations more frequently occurred within the first 3 months after the diagnosis of advanced cancer, and in the last 3 months before death. A short - less than 3 months - delay between diagnosis and unplanned hospitalization was associated with poor outcome. 272 (88%) patients received optimized SCC, within a median delay of 8 weeks after diagnosis. Intervention of the nurse for in- and out-patient network coordination was done for 143 (46%) patients, and at-home hospitalization was organized for 78 (25%) patients. The outcome of patients who received optimized SCC was numerically, but not significantly better (median overall survival of 11.8 vs. 6.9 months, p = 0.270).Conclusion: Our study provides landmark data to support an early integration of optimized SCC for patients with advanced lung cancer, that includes multimodal supportive care interventions along the course of the disease. This highlights the role of multidisciplinary teams to optimize the management of patients with advanced lung cancer

Profils aérodynamiques de patients dysodiques : une étude de cas

International audienceLa voix chantée sollicite parfois de façon extrême l’appareil vocal selon le style de chant, les différentes pratiques et le niveau d’entraînement (Aronson et Bless, 2009). Lorsqu’elle est altérée, on parle de dysodie (perte des habiletés vocales chantées, associée ou non à une laryngopathie), se traduisant par des signes acoustiques (anomalies tonales, dynamiques ou de timbre) et/ou physiques. Elle peut avoir un retentissement psychologique (Amy de la Bretèque, 2012). Les patients consultant pour dysodie sont des chanteurs amateurs, professionnels ou étudiants. Les chanteurs sont plus enclins à développer des troubles vocaux (Miller et Verdolini, 1995). La grande majorité des dysodiques sont des femmes (Hunter et al., 2011). Le nodule reste la lésion la plus retrouvée mais les lésions congénitales arrivent en second chez les chanteurs (Bouchayer et Cornut, 1992).L’intérêt des paramètres aérodynamiques pour le diagnostic des pathologies vocales est aujourd’hui largement démontré (Holmberg et al., 2003). Parmi eux, les mesures de pression sous-glottique estimée (PSGE) et de débit d’air oral (DAO) dépendent du niveau d’entraînement des chanteurs et du type de dysfonction vocale. Ces paramètres sont plus élevés chez les chanteurs lyriques que chez les non chanteurs (Dargin et Searl, 2015). En clinique, la PSGE est plus élevée lors du forçage vocal (Morsomme et al., 2015). Concernant le DAO, les résultats sont controversés : augmentation avec l’effort vocal chez des non chanteurs euphoniques (Rosenthal et al. 2014) ; diminution lors de l’augmentation d’intensité chez un non chanteur euphonique (Pillot-Loiseau, 2011).Nous avons étudié le comportement aérodynamique de 6 chanteuses et 4 chanteurs venus consulter en phoniatrie, puis les avons comparés à une population euphonique (13 chanteuses et 6 chanteurs) recrutée lors d’une précédente étude (Beaud, 2015). Les profils de PSGE et de DAO selon la fréquence montrent une très grande variabilité interindividuelle (Figures 1 à 4). Chez les patients dysodiques (Figures 1 et 4), ils corroborent les observations cliniques. L’exemple d’une patiente (P4), chanteuse d’opéra professionnelle avec sulcus, montre une corrélation entre sa plainte (difficultés d’aigus depuis le début de son activité), les données cliniques (difficulté d’initialisation du son au-delà du ré4) et les données aérodynamiques : le contrôle des débits et pressions est correct jusqu’au do4 (554Hz) environ. Au-delà, ces valeurs diminuent soudainement. Chez une autre chanteuse (P8), choriste amateur et enseignante avec nodules, le bilan vocal décrit une dysphonie modérée et une désonorisation lors de la transition entre mécanismes laryngés M1 et M2. Les données aérodynamiques reflètent le dysfonctionnement : le débit est anormalement bas, les valeurs de pression sont augmentées et leur évolution avec la hauteur n’est pas régulière. On retrouve une zone d’instabilité autour de 300Hz (zone de passage) : la pression et le débit diminuent puis ré-augmentent. Ces comportements s’éloignent de ceux des chanteurs euphoniques chez qui la pression augmente régulièrement avec la hauteur et l’intensité (Figures 2 et 3). Ces difficultés reflétées dans la gestion aérodynamique des notes chantées appellent à une rééducation ciblée sur le comportement pneumophonique dans le chant, pour laquelle la méthode de rééducation à la paille pourrait être prometteuse

Risk of Anticoagulant Rodenticide Exposure for Mammals and Birds in Parc National des Pyrénées, France

International audienceThe extensive use of anticoagulant rodenticides (ARs) to control rodent populations poses intoxication risks for wildlife: persistence of ARs in rodents can cause secondary exposure and poisoning of predators or scavengers. We investigated risk factors for wildlife exposure to ARs in the Parc National des Pyrenees (PNP), France, using a multivariable logistic regression analysis. A total of 157 liver samples were collected from carcasses of 10 mammal and three bird species found in the PNP between 2010 and 2018 and screened for presence of AR residues. First- and second-generation ARs were detected in more than 60% of red fox (Vu/pes vulpes) and stone marten (Martes foina) samples and in around 40% of wild cat (Fells silvestris), European pine marten (Martes martes), American mink (Neovison vison), and Eurasian Buzzard (Buteo buteo) samples. Wildlife exposure to ARs was significantly associated with species having a regular consumption of small mammals (odds ratio [OR]: 2.5, 95% confidence interval [CI]: 1.1-5.8) being collected in the Ossau valley (OR: 2.5, 95% CI: 1.1-6.1) and between 2013 and 2015 (OR: 4.8, 95% CI: 2.0-11.7). We identified wild species that could be targeted for risk-based surveillance program for AR secondary exposure and determined high risk areas in which alternative measures should be applied for rodent control

Probing the functions of carbohydrate binding Modules in the CBEL protein from the oomycete Phytophthora parasitica

Oomycetes are microorganisms that are distantly related to true fungi and many members of this phylum are major plant pathogens. Oomycetes express proteins that are able to interact with plant cell wall polysaccharides, such as cellulose. This interaction is thought to be mediated by carbohydrate-binding modules that are classified into CBM family 1 in the CAZy database. In this study, the two CBMs (1-1 and 1-2) that form part of the cell wall glycoprotein, CBEL, from Phytophthora parasitica have been submitted to detailed characterization, first to better quantify their interaction with cellulose and second to determine whether these CBMs can be useful for biotechnological applications, such as biomass hydrolysis. A variety of biophysical techniques were used to study the interaction of the CBMs with various substrates and the data obtained indicate that CBEL's CBM1-1 exhibits much greater cellulose binding ability than CBM1-2. Engineering of the family 11 xylanase from Talaromyces versatilis (TvXynB), an enzyme that naturally bears a fungal family 1 CBM, has produced two variants. The first one lacks its native CBM, whereas the second contains the CBEL CBM1-1. The study of these enzymes has revealed that wild type TvXynB binds to cellulose, via its CBM1, and that the substitution of its CBM by oomycetal CBM1-1 does not affect its activity on wheat straw. However, intriguingly the addition of CBEL during the hydrolysis of wheat straw actually potentiates the action of TvXynB variant lacking a CBM1. This suggests that the potentiating effect of CBM1-1 might not require the formation of a covalent linkage to TvXynB

Understanding EU policies and the EU Green Deal:A policy mapping and scoping of institutional barriers within EU governance

The PermaGov Deliverable focuses on exploring the EU policy landscape within the context of the European Green Deal (EGD), structured around four regime complexes: marine life, marine plastics, marine energy, and maritime transport. These complexes provide a framework for analysing the EU's approach to achieving the EGD's vision for sustainable marine governance. This report aims to offer a descriptive overview of marine EU policies relevant to the PermaGov project, focusing on policies identified as relevant to the overarching goals set forth in the EGD. It also considers relevant initiatives at global and regional levels.The marine life regime sees the EU Biodiversity Strategy for 2030 as its overarching strategy, essential for the EGD’s element of preserving and restoring ecosystems and biodiversity. Tackling the challenges of marine waste pollution, the marine plastics regime is guided by the EU Circular Economy Action Plan and the EU Action Plan: Towards Zero Pollution for Air, Water, and Soil, targeting the EGD’s elements of a mobilising industry for a clean and circular economy and a zero-pollution ambition for a toxic-free environment. The marine energy regime is shaped by the European Climate Law and the Offshore Renewable Energy Strategy, which are the overarching instruments that contribute to the EGD’s elements of increase the EU’s climate ambition for 2030 and 2050 and ensure the supply of clean, affordable, and secure energy. Lastly, the maritime transport regime sees the'Fit for 55'Package and the'Sustainable and Smart Mobility Strategy'as the two main instruments to achieve the EGD’s elements of increase the EU

Characterization of CBEL binding to cellulose nanocrystals by fluorescence spectroscopy.

<p>N.D: no binding detected.</p><p>Characterization of CBEL binding to cellulose nanocrystals by fluorescence spectroscopy.</p

Characterization of CBEL and <i>Tv</i>XynB binding to Avicel by solid state depletion assay.

<p>¹ Bmax is expressed in μmol of protein per g of substrate.</p><p>²<i>K</i>_D is expressed as μmol.L^-1 of proteins.</p><p>Characterization of CBEL and <i>Tv</i>XynB binding to Avicel by solid state depletion assay.</p

Schematic representation of CBEL protein domains.

<p>CBM1s and PAN/Apple are numbered from the N- to the C-terminus of the protein. CBM1s and PAN/Apple domains are symbolized by grey and white boxes respectively. The linker is represented by a black line.</p