Suchergebnisse

As bridges provide crossing at critical points such as waterways, valleys and other type of physical obstacles, they are invaluable assets within a rail network. However, these infrastructures are exposed to several hazards of different natures that compromise their structural safety and ultimately lead to critical events. Accord-ing to literature, there are commonly five levels of assessment of safety with increasing complexity per level. Level 1, consisting on an assessment based on partial coefficients, is the simplest one, whereas level 5 is the most complex involving a combination of non-linear analysis and probabilistic assessment. Within that scope, this paper analyses the structural safety of a pre-stressed reinforced concrete railway bridge located in Portugal, by using a combination of linear elastic and probabilistic analysis procedures. The proposed framework is adaptable and suitable for common safety analysis. The obtained reliability index is compared to threshold values given by different standards which validates that the bridge ensures the safety at ultimate limit state. The reliability levels along time are also determined assuming a reduction in the cross-section area of the steel reinforcement, resulting from corro-sion initiation based on onsite inspection evidences. From the analysis of this scenario over time, it may be concluded that the bridge under study exhibits sufficient load carrying capacity during its planned service life even under the proposed damage. ; The third author would like to thank FCT – Portuguese Scientific Foundation for the research grant PD/BD/128015/2016 under the PhD program "Innovation in Railway System and Technologies- iRail". This work was partly financed by FEDER funds through the Competitivity Factors Operational Programme - COMPETE and by national funds through FCT Foundation for Science and Technology within the scope of the project POCI-01- 0145-FEDER-007633. This project has received funding from the European Union's Horizon 2020 research and innovation ...

A study of Magnetite (Fe3O4) as a suitable matrix for the improved adhesion and proliferation of MC3T3-E1 pre-osteoblast cells in bone regeneration is presented. Biodegradable and magnetic polycaprolactone (PCL)/magnetite (Fe3O4) scaffolds, which were fabricated by Thermally Induced Phase Separation, are likewise analyzed. Various techniques are used to investigate in vitro degradation at 37 °C, over 104 weeks, in a phosphate buffered saline (PBS) solution. Magnetic measurements that were performed at physiological temperature (310 K) indicated that degradation neither modified the nature nor the distribution of the magnetite nanoparticles. The coercive field strength of the porous matrices demonstrated ferromagnetic behavior and the probable presence of particle interactions. The added nanoparticles facilitated the absorption of PBS, with no considerable increase in matrix degradation rates, as shown by the Gel Permeation Chromatography (GPC) results for Mw, Mn, and I. There was no collapse of the scaffold structures that maintained their structural integrity. Their suitability for bone regeneration was also supported by the absence of matrix cytotoxicity in assays, even after additions of up to 20% magnetite. ; This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry Department under the ELKARTEK, HAZITEK and PIBA programs. Supported by the Portuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding UID/FIS/04650/2013, project POCI-01-0145-FEDER-028237 and grant SFRH/BD/111478/2015 (S.R.) is acknowledged. ; Technical and human support provided by SGIker (UPV/EHU, MICINN, GV/EJ, ERDF and ESF) is gratefully appreciated. The authors acknowledge funding by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry and Education Department under the ...

The use of mechanical anchorage systems can delay or avoid premature failure of reinforced concrete (RC) structures with externally-bonded fiber-reinforced polymer (EB-FRP) laminates. This work reports the results of an experimental program aimed at studying the bond behavior of a metallic anchorage plate, typically used for pre-stressed EB-FRP systems. The overall experimental program comprised 17 concrete prisms with carbon fiber reinforced polymer (CFRP) laminates externally bonded to the concrete with mechanical anchorage at the end and where different levels of normal stress were applied. Direct shear tests were carried out using two configurations: (i) the steady-state tests, where the laminate was pulled out from the block with increasing force and constant temperature (20 ºC, 60 ºC and 80 ºC); (ii) and the transient tests, where the laminate was pulled out with constant force (0.36%, 0.45% and 0.54% of strain) and the temperature was gradually increased. Experimental results showed that the ultimate capacity of the mechanical anchorage can decrease by 44-59% depending on the temperature and level of normal stress. ; This work was supported by FEDER funds through the Operational Program for Competitiveness Factors – COMPETE and National Funds through FCT (Portuguese Foundation for Science and Technology) under the project FRPLongDur POCI-01-0145- FEDER-016900 (FCT PTDC/ECM-EST/1282/2014) and partly financed by the project POCI544 01-0145-FEDER-007633. The first author wishes also to acknowledge the grant SFRH/BD/98309/2013 provided by FCT. The second author acknowledges the Spanish Government (Ministerio de Educación, Cultura y Deporte) for the grant José Castillejo ref. CAS16/00288. Finally, the authors also like to thank all the companies that have been involved supporting and contributing for the development of this study, mainly: the S&P Clever Reinforcement Ibérica Lda. company for providing the laminates, the epoxy adhesive and metallic anchorage plates; the Hilti Portugal-Produtos e Serviços, ...

In order to massify energy improvement of residential existing buildings, it's important to take into account the complex criteria defining the housing stock and to consider various scales of intervention. To reduce the energy consumption of this sector, it's necessary to focus the action where energy saving is most important. Our ambition consists in developing methods allowing to exploit different residential sector databases in order to generate accuracy energy diagnosis at building scale first, to aggregate them at a higher scale level then. This approach is based on data series implementation in a GIS tool combined with a physical approach.Indeed, since GIS is not only intended for producing a map, it's possible to exploit its ability to manage spatial data and to produce geometric parameters necessary for energy calculations. These parameters can be enriched or crossed with other data (such as the age of the building, envelope components, climate) to estimate energy consumption and to forecast demand.Building geometry description is necessary to estimate energy heat balance. The geometrical envelope characteristics can be extracted from the information of building entities present in the BD TOPO spatial database. Some procedures were developed and validated to evaluate the areas of different envelope components: area of external walls, roofs, floors, volume and identify adjoining building surfaces. All these parameters allowed us to calculate the precise orientation of each elevation and to evaluate real solar gains.The second database used corresponds to data gathered, treated and structured by CEREMA from the complex taxation files of government. Among the different data collection, this database contains information about age, dwelling number and habitable area aggregated at the parcel. This database allowed us to identify archetypes. The TABULA nomenclature was chosen to be used in the framework of ATRE project because it's an exhaustive national classification. The spatial data can be crossed with ...

Free PMC Article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6242839/ ; Anthrax is an infectious disease caused by Bacillus anthracis, a bioterrorism agent that develops resistance to clinically used antibiotics. Therefore, alternative mechanisms of action remain a challenge. Herein, we disclose deoxy glycosides responsible for specific carbohydrate-phospholipid interactions, causing phosphatidylethanolamine lamellar-to-inverted hexagonal phase transition and acting over B. anthracis and Bacillus cereus as potent and selective bactericides. Biological studies of the synthesized compound series differing in the anomeric atom, glycone configuration and deoxygenation pattern show that the latter is indeed a key modulator of efficacy and selectivity. Biomolecular simulations show no tendency to pore formation, whereas differential metabolomics and genomics rule out proteins as targets. Complete bacteria cell death in 10 min and cellular envelope disruption corroborate an effect over lipid polymorphism. Biophysical approaches show monolayer and bilayer reorganization with fast and high permeabilizing activity toward phosphatidylethanolamine membranes. Absence of bacterial resistance further supports this mechanism, triggering innovation on membrane-targeting antimicrobials. ; The European Union is gratefully acknowledged for the support of the project "Diagnostic and Drug Discovery Initiative for Alzheimer's Disease" (D3i4AD), FP7-PEOPLE-2013-IAPP, GA 612347. We thank the Management Authorities of the European Regional Development Fund and the National Strategic Reference Framework for the support of the Incentive System - Research and Technological Development Co-Promotion FACIB Project number 21457. Fundação para a Ciência e a Tecnologia is also acknowledged for the support of projects UID/Multi/00612/2013, FCT/UID/ Multi/04046/2013, IF/00808/2013/CP1159/CT0003, PTDC/BBBBQB/6071/2014, as well as for the post-doc grant SFRH/BPD/42567/2007 (A.M.), the Ph.D. grants SFRH/BDE/51998/2012 (C.D.), and SFRH/BDE/51957/2012 (J.P.P.), both co-sponsored by CIPAN, and also for the Ph.D. grant SFRH/BD/116614/2016 (R.N.). ; info:eu-repo/semantics/publishedVersion

This study aimed to assess the bioaccessibility of different marine biotoxins in naturally contaminated shellfish and fish gonads using an in vitro digestion methodology. In general, hydrophilic toxins (domoic acid, paralytic shellfish poisoning toxins and tetrodotoxins) showed higher bioaccessibility than lipophilic ones (okadaic acid and azaspiracids). The bioaccessibility of toxins from the okadaic acid group ranged from 69% (raw European razor clams) to 74% (raw donax clams). Regarding azaspiracids, 47% of the initial content was bioaccessible in steamed blue mussel. As for hydrophilic toxins, 100% of the initial content was bioaccessible after digestion in raw shellfish and puffer fish gonads. The total tetrodotoxin bioaccessibility in puffer fish gonads decreased significantly after steaming. The profile of tetrodotoxins changed during the digestion process: TTX and 11-norTTX-6S-ol analogues decreased significantly after digestion, but the 5,6,11-trideoxy TTX analogue increased in both raw and steamed puffer fish gonads. These preliminary findings confirm the need to consider bioaccessibility data in future seafood risk assessment, as such information enables a more accurate and realistic estimation of potential seafood hazards, particularly in what concerns lipophilic toxins, therefore, constituting a crucial tool in the refinement of regulatory limits for the presence of biotoxins in seafood. ; The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under the ECsafeSEAFOOD project (grant agreement n° 311820) and through the European Regional Development Fund. IRTA members acknowledge support from CERCA Programme/Generalitat de Catalunya. L.R. acknowledges a scholarship from the ECsafeSEAFOOD project. Ana C. Braga has a Doctoral Grant (PD/BD/113484/2015) from the Portuguese Science and Technology Foundation (FCT), whereas Antonio Marques and Pedro R. Costa are supported through the FCT Investigator program (IF). Lagocephalus sceleratus was kindly provided by Andrés Izquierdo-Muñoz from Centro de Investigación Marina de Santa Pola (CIMA) (Spain).

Polyoxyethanyl--tocopheryl sebacate (PTS) is an amphiphilic compound with self-emulsifying properties known to form micelles. In this work, we report the production of PTS micelles for the encapsulation and delivery of a hydrophobic derivative of methotrexate, MTX di-ethylated (MTX-OEt). We optimized the micelles production by testing two different techniques: auxiliary solvent and sonication. Small and homogeneous micelles ( 40 nm) were obtained through the auxiliary solvent method performed at 30 °C and using 15 mg/mL of PTS. The produced micelles with the most promising physicochemical properties did not induce cytotoxicity when tested in normal human cells (BJ5ta cells), being considered for the encapsulation of MTX-OEt. This prodrug was achieved by Fisher esterification using ethanol, being isolated in good yield (= 68 %). MTX-OEt was efficiently encapsulated onto the produced micelles which preserved their physicochemical properties. The PTS micelles loaded with MTX-OEt, free MTX-OEt and free unmodified MTX revealed similar biological effect against cancer cells (Caco-2 cells). These results demonstrated that the biological activity of MTX is not altered after modification. The developed PTS micelles revealed a promising intracellular delivery performance with potentiality for cancer therapy. ; This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/ BIO/04469/2019 unit and BioTecNorte operation (NORTE-01-0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. The authors thanks to FCT for funding their scholarship: Jennifer Noro (SFRH/BD/121673/2016) and Carla Silva (SFRH/IF/00186/ 2015). This work has also received funding from the European Union Horizon 2020 research and innovation program under grant agreement NMP-06-2015-683356 FOLSMART. ...

Tendon-to-bone interfaces exhibit a hierarchical multitissue transition. To replicate the progression from mineralized to nonmineralized tissue, a novel 3D fibrous scaffold is fabricated with spatial control over mineral distribution and cellular alignment. For this purpose, wet-spun continuous microfibers are produced using polycaprolactone (PCL)/ gelatin and PCL/gelatin/hydroxyapatite nano-to-microparticles (HAp). Higher extrusion rates result in aligned PCL/gelatin microfibers while, in the case of PCL/gelatin/HAp, the presence of minerals leads to a less organized structure. Biological performance using human adipose-derived stem cells (hASCs) demonstrates that topography of PCL/gelatin microfibers can induce cytoskeleton elongation, resembling native tenogenic organization. Matrix mineralization on PCL/gelatin/HAp wet-spun composite microfibers suggests the production of an osteogenic-like matrix, without external addition of osteogenic medium supplementation. As proof of concept, a 3D gradient structure is produced by assembling PCL/gelatin and PCL/gelatin/HAp microfibers, resulting in a fibrous scaffold with a continuous topographical and compositional gradient. Overall, the feasibility of wet-spinning for the generation of continuously aligned and textured microfibers is demonstrated, which can be further assembled into more complex 3D gradient structures to mimic characteristic features of tendon-to-bone interfaces. ; The authors thank to Hospital da Prelada (Porto, Portugal) for providing lipoaspirate tissue (Plastic Surgery Department) samples; Dr. Rui Domingues and Dr. Pedro Babo for the technical help with confocal microscopy and micro-CT analysis. The authors acknowledge the financial support from the European Union Framework Programme for Research and Innovation HORIZON2020, under the TEAMING Grant agreement No 739572 – The Discoveries CTR, the ERC Grant CoG MagTendon nr 772817, FCT-Fundação para a Ciência e a Tecnologia for the PhD grant of IC (PD/BD/128088/2016); and the Project ...

BACKGROUND: Biocides play an integral role in infection control. Paralleling concern about rising incidence of multidrug-resistant (MDR) organisms is a concern for resistance to biocides. In small studies, several genes involved in the production of efflux pump proteins have been identified as markers of biocide resistance in Klebsiella spp., namely cepA, qacA, qacE, qac∆E, and acrA. This study aimed to analyze the Klebsiella spp. isolates of a previously defined military trauma group with a high incidence of MDR organisms for the presence of these genes and their correlation with other resistance. METHODS: All infecting K. pneumoniae, K. variicola, and K. quasipneumoniae isolates archived by the Trauma Infectious Disease Outcomes Study (June 2009–December 2014) were selected. Additionally, all colonizing isolates linked with infecting isolates were included; the remainder to total 50 MDR and 46 non-MDR colonizing isolates were chosen randomly. Antimicrobial identification and susceptibilities were determined by CLSI criteria using the BD Phoenix Automated Microbiology System. PCR according to published methods for cepA, qacA, qacE, qac∆E, and acrA was accomplished in duplicate. MDR was defined as either resistance to ≥3 classes of aminoglycosides, β-lactams, carbapenems and/or fluoroquinolones or production of an ESBL or KPC. RESULTS: A total of 237 isolates (221 K. pneumoniae, 10 K. variicola, 6 K. quasipneumoniae) met inclusion criteria, of which 149 (63%) were MDR. All isolates had been exposed to antimicrobials prior to isolation. Of all isolates, 234 (98%) carried cepA: 218 (98%) K. pneumoniae carried cepA, 10 (100%) K. variicola carried cepA, and 6 (100%) of K. quasipneumoniae carried cepA. In addition, 148 (62%) isolates with cepA were MDR. One (10%) K. variicola isolate carried qacE along with cepA. This isolate was the only MDR K. variicola. None of the isolates carried qacA, qac∆E, or acrA. CONCLUSION: We confirmed the near universal presence of the cepA biocide resistance gene in Klebsiella spp. ...

scRNA-seq data for this project has been deposited in ArrayExpress under the accession numbers E-MTAB-6072 and E-MTAB-7311. Processed data can be found in https://figshare.com/projects/Treg_scRNA-seq/38864, and analysis notebooks can be found in https://github.com/tomasgomes/Treg_analysis. ; Non-lymphoid tissues (NLTs) harbor a pool of adaptive immune cells with largely unexplored phenotype and development. We used single-cell RNA-seq to characterize 35,000 CD4+ regulatory (Treg) and memory (Tmem) T cells in mouse skin and colon, their respective draining lymph nodes (LNs) and spleen. In these tissues, we identified Treg cell subpopulations with distinct degrees of NLT phenotype. Subpopulation pseudotime ordering and gene kinetics were consistent in recruitment to skin and colon, yet the initial NLT-priming in LNs and the final stages of NLT functional adaptation reflected tissue-specific differences. Predicted kinetics were recapitulated using an in vivo melanoma-induction model, validating key regulators and receptors. Finally, we profiled human blood and NLT Treg and Tmem cells, and identified cross-mammalian conserved tissue signatures. In summary, we describe the relationship between Treg cell heterogeneity and recruitment to NLTs through the combined use of computational prediction and in vivo validation. ; Wethank V. Proserpio, M. Stubbington, T. Hagai, F.V. Braga, V. Svensson, and J. Henriksson for helpful discussions and advice; T. Hagai, R. Vento-Tormo, K. Meyer, and J. Park for critical reading of the manuscript; and B. Koppelman for editorial support. We thank WSI Single-cell Genomic Core Facility, WSI Sequencing Facility, WSI Flow Cytometry facilities, K. Polanski, and the Kennedy Institute Flow Cytometry Facility for expert technical advice and assistance. R.J.M. was supported by a PhD Fellowship from the Fundação para a Ciência e Tecnologia, Portugal (SFRH/BD/51950/2012) and T.G. by the European Union's H2020 research and innovation program ''ENLIGHT-TEN'' under the Marie Sklodowska-Curie grant ...

Turkey is a member of the Council of Europe, of the North Atlantic Treaty Organization, and a candidate to the accession to the European Union. As a transcontinental country, Turkey belongs both to Asia and Europe, and can be used as a benchmark to assess the narrowing of human rights and civil liberties in the fight against terrorism. While having become the central platform for action for Islamist groups in the Middle East, as a result of the gradually Islamized domestic and foreign policy of Ankara since 2011, Turkey demands its Western allies to stand by it in the fight against terrorism. Turkey has a concept of terrorism that clashes with the principles enshrined in the European Convention on Human Rights, that should be obliged to comply, together with other international instruments. The authoritarian turn put in place by the Turkish government after the failed coup d'état of 15 July 2016 is a challenge to the values of European and Western civilization, and pillories all the contradictions and the limits of the latter concerning defending fundamental human rights while countering terrorism. This paper scrutinizes the policy of the Turkish authorities to counter terrorism within the country and at its external borders, and the inconsistencies of this policy with the international legal commitments and with the values of European and Western nations. ; Abstract of the paper presentation published in the Conference Programme & Abstract Book of the 6th International Conference on Eurasian Politics and Society (IEPAS2019), held at the Universidade Autónoma de Lisboa (UAL), Portugal, on 4-5 July 2019. The author gratefully acknowledges the European Social Fund (ESF) and the Fundação para a Ciência e a Tecnologia (FCT), Portugal, for supporting this research through grant SFRH/BD/136170/2018.

Threats from Africa undermine the security of Europe. The European Union and NATO have 22 member countries in common, namely most of EU member states. The fundamental role of the Atlantic Alliance is to safeguard the freedom and security of its member countries by political and military means. Since the fall of the Berlin Wall, non-state actors and hybrid threats emerged as the major challenges to international security. NATO is evolving in response to new strategic reality, and terrorism and weak states are among the most pressing challenges the Alliance faces. NATO is playing an increasingly important role in crisis management and peacekeeping. Since the African Union was established in 2001 as the successor to the Organization of African Unity,it faces difficulties in achieving the African ownership in peace and security. The chronic lack of funding and of appropriate military resources, makes the African Union depending on external aid –NATO provides military support, while the EU provides money. NATO cooperates with the AU since 2005, and all NATO-AU activities are coordinated with the EU. The NATO-EU partnership is complementar and mutual, and is based on common values and strategic interests, and should be strengthened to secure the European southern border, and to guarantee security in the Mediterranean region. Therefore, stronger and permanent NATO–EU partnership in Africa would be desirable. This brief paper investigates the ties between these three organizations, and suggests some recommendations to improve the 'trilateral' relationship to ensure security to their member states. ; The author gratefully acknowledges the European Social Fund (ESF) and the Fundação para a Ciência e a Tecnologia (FCT), Portugal, for supporting this publication through grant SFRH/BD/136170/2018, and the Knights of Vartan Fund for Armenian Studies administered by the National Association for Armenian Studies and Research (NAASR). This working paper is published at https://iep.lisboa.ucp.pt/pt-pt/asset/5121/file

Tetraspanins, such as CD81, can form lateral associations with each other and with other transmembrane proteins. These interactions may underlie CD81 functions in multiple cellular processes, such as adhesion, morphology, migration, and differentiation. Since CD81's role in neuronal cells' migration has not been established, we here evaluated effects of CD81 on the migratory phenotype of SH-SY5Y neuroblastoma cells. CD81 was found enriched at SH-SY5Y cell's membrane, co-localizing with its interactor filamentous-actin (F-actin) in migratory relevant structures of the leading edge (filopodia, stress fibers, and adhesion sites). CD81 overexpression increased the number of cells with a migratory phenotype, in a potentially phosphatidylinositol 3 kinase (PI3K)-Ak strain transforming (AKT) mediated manner. Indeed, CD81 also co-localized with AKT, a CD81-interactor and actin remodeling agent, at the inner leaflet of the plasma membrane. Pharmacologic inhibition of PI3K, the canonical AKT activator, led both to a decrease in the acquisition of a migratory phenotype and to a redistribution of intracellular CD81 and F-actin into cytoplasmic agglomerates. These findings suggest that in neuronal-like cells CD81 bridges active AKT and actin, promoting the actin remodeling that leads to a motile cell morphology. Further studies on this CD81-mediated mechanism will improve our knowledge on important physiological and pathological processes such as cell migration and differentiation, and tumor metastasis. ; This work was supported by Fundação para a Ciência e Tecnologia (Portuguese Ministry of Science and Technology), Centro 2020 and Portugal2020, the COMPETE program, QREN, and the European Union (FEDER program) via the Institute for Biomedicine iBiMED UID/BIM/ 04501/2013, fellowship SFRH/BD/90996/2012, project PTDC/CVT-CVT/ 32261/2017, and the support of the LiM facility of iBiMED, a member of the Portuguese Platform of BioImaging (PPBI- POCI-01-0145-FEDER-022122). ; published

Polymeric ion selective electrodes are highly sensitive to changes in zero current ion flow and this offers a route to signal amplification in label-free potentiometric immunosensors. In this work, a label-free potentiometric immunosensor toward Salmonella typhimurium (ST) assembled in a home-made pipette-tip electrode is described. The signal-output amplification was implemented on a gold nanoparticle polymer inclusion membrane (AuNPs-PIM) which was used as sensing platform and for antibody immobilization. Additionally, a marker ion was used to detect the antibody-antigen binding event at the electrode surface. The immunosensor construction was performed in several steps: i) gold salt ions extraction in PVC membrane; ii) AuNPs formation using Na2EDTA as reduction agent; iii) antibody anti-Salmonella conjugation on AuNPs-PIM in pipette-tip electrodes. The potential shift observed in potentiometric measurements was derived simply from the blocking effect in the ionic flux caused by antigen-antibody conjugation, without no extra steps, mimetizing the ion-channel sensors. A detection limit of 6 cells mL-1 was attained. As proof-of-concept, recovery studies were performed in spiked commercial apple juice samples with success. Due to the simplicity of use, the appealing cost of equipment and sensor production and being able to provide a quick analytical response (less than 1 h for a complete assay, including sample preparation for analysis), this scheme represents a good prototype device for the detection of foodborne pathogens like ST or other immune-responsive bacteria. ; This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds from Portugal; FCT-Fundação para a Ciência e a Tecnologia through projects UID/QUI/50006/2013 and Norte-01-0145-FEDER-000011-RL1-QUALIFOOD. N.F.D Silva and M.F. Barroso are gratefully to FCT grant SFRH/BD/112414/2015 and SFRH/BPD/78845/2011, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by FSE and MCTES. ; info:eu-repo/semantics/publishedVersion

Cytoplasmic dynein 1 (dynein) is the predominant microtubule minus end-directed motor in animals and participates in a wide range of cellular processes, including membrane trafficking, nuclear migration, and cell division. Dynein's functional diversity depends on co-factors that regulate its subcellular localization, interaction with cargo, and motor activity. The ubiquitous co-factor nuclear distribution gene E (NudE) is implicated in many of dynein's functions, and mutations in NudE cause the brain developmental disease microcephaly. To identify genetic interactors of the Caenorhabditis elegans NudE homolog nud-2, we performed a genome-wide RNAi screen with the null allele nud-2(ok949), which compromises dynein function but leaves animals viable and fertile. Using bacterial feeding to deliver dsRNAs in a 96-well liquid format and a semi-automated fluorescence microscopy approach for counting parents and progeny, we screened 19762 bacterial clones and identified 38 genes whose inhibition caused enhanced lethality in nud-2(ok949) relative to the nud-2(+) control. Further study of these genes, many of which participate in cell division, promises to provide insight into the function and regulation of dynein. ; The authors acknowledge the support of the BioSciences Screening i3S Scientific Platform. Funding for this project was provided by the European Research Council under the European Union's Seventh Framework Programme (ERC grant agreement n o ERC-2013-StG-338410-DYNEINOME to R.G.), by the European Molecular Biology Organization (EMBO Installation Grant 2545 to R.G.), by the Fundacao para a Ciencia e a Tecnologia (IF/01015/2013/CP1157/CT0006 to R.G. and SFRH/BD/103495/2014 to H. R.), and by 'Norte-01-0145-FEDER-000029-Advancing cancer research: from basic knowledge to application', supported by the Norte Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement through the European Regional Development Fund (FEDER)