RESUMEN
Core facilities have a different mission than academic research labs. Accordingly, they require different career paths and structures.
RESUMEN
Core facilities play a central role in the life sciences by generating data and ensuring quality standards. Their contributions to research should be appropriately acknowledged or cited in research papers.
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Bibliometría , Disciplinas de las Ciencias Biológicas , PublicacionesRESUMEN
Core facilities offer visiting scientists access to equipment and expertise to generate and analyze data. For some projects, it might however be more efficient to collaborate remotely by sending in samples.
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Servicios PostalesRESUMEN
A series of substituted benzylsulfanyl-phenylamines was synthesized, of which four substituted benzylsulfanyl-phenylguanidines (665, 666, 667 and 684) showed potent fungicidal activity (minimal fungicidal concentration, MFC ≤ 10 µM for Candida albicans and Candida glabrata). A benzylsulfanyl-phenyl scaffold with an unsubstituted guanidine resulted in less active compounds (MFC=50-100 µM), whereas substitution with an unsubstituted amine group resulted in compounds without fungicidal activity. Compounds 665, 666, 667 and 684 also showed activity against single C. albicans biofilms and biofilms consisting of C. albicans and Staphylococcus epidermidis (minimal concentration resulting in 50% eradication of the biofilm, BEC50 ≤ 121 µM for both biofilm setups). Compounds 665 and 666 combined potent fungicidal (MFC=5 µM) and bactericidal activity (minimal bactericidal concentration, MBC for S. epidermidis ≤ 4 µM). In an in vivo Caenorhabditis elegans model, compounds 665 and 667 exhibited less toxicity than 666 and 684. Moreover, addition of those compounds to Candida-infected C. elegans cultures resulted in increased survival of Candida-infected worms, demonstrating their in vivo efficacy in a mini-host model.
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Antifúngicos/síntesis química , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Guanidinas/síntesis química , Guanidinas/farmacología , Animales , Antifúngicos/química , Antifúngicos/toxicidad , Caenorhabditis elegans/efectos de los fármacos , Guanidinas/química , Guanidinas/toxicidad , Modelos Biológicos , Estructura Molecular , Sulfuros/síntesis química , Sulfuros/química , Sulfuros/farmacología , Sulfuros/toxicidadRESUMEN
To unravel the working mechanism of the fungicidal piperazine-1-carboxamidine derivative BAR0329, we found that its intracellular accumulation in Saccharomyces cerevisiae is dependent on functional lipid rafts. Moreover, BAR0329 induced caspase-dependent apoptosis in yeast, in which the mitochondrial fission machinery consisting of Fis1 (Whi2), Dnm1 and Mdv1 is involved. Our data are consistent with a prosurvival function of Fis1 (Whi2) and a proapoptotic function of Dnm1 and Mdv1 during BAR0329-induced yeast cell death.
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Antifúngicos/toxicidad , Apoptosis , Mitocondrias/efectos de los fármacos , Piperazinas/toxicidad , Saccharomyces cerevisiae/efectos de los fármacos , Proteínas Adaptadoras Transductoras de Señales/metabolismo , GTP Fosfohidrolasas/metabolismo , Microdominios de Membrana/metabolismo , Proteínas Mitocondriales/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
Herpesviruses have developed different tools to thwart efficient antibody-dependent neutralisation and lysis of virions and elimination of infected cells. This overview will briefly summarize different of these tools, including (i) viral Fc receptors and the resulting process of antibody bridging, (ii) internalization of individual viral proteins and clustered antibody-antigen complexes from the plasma membrane of infected cells, and (iii) directed egress of virus particles to sites of intimate cell-cell contact that are difficult to access for antibodies.
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Anticuerpos Antivirales/inmunología , Herpesviridae/inmunología , Proteínas del Envoltorio Viral/inmunología , Animales , Complejo Antígeno-Anticuerpo/inmunología , Antígenos Virales/inmunología , Comunicación Celular/inmunología , Pruebas Inmunológicas de Citotoxicidad , Glicoproteínas de Membrana/inmunología , Receptores Fc/inmunologíaRESUMEN
Translational research leaves no-one indifferent and everyone expects a particular benefit. We as EU-LIFE (www.eu-life.eu), an alliance of 13 research institutes in European life sciences, would like to share our experience in an attempt to identify measures to promote translational research without undermining basic exploratory research and academic freedom.
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Disciplinas de las Ciencias Biológicas/organización & administración , Investigación Biomédica/organización & administración , Investigación Biomédica Traslacional/organización & administración , Disciplinas de las Ciencias Biológicas/tendencias , Investigación Biomédica/tendencias , Europa (Continente) , Humanos , Cooperación Internacional , Investigación Biomédica Traslacional/tendenciasRESUMEN
Two major structural elements of a cell are the cytoskeleton and the lipid membranes. Actin and cholesterol are key components of the cytoskeleton and membranes, respectively, and are involved in a plethora of different cellular processes. This review summarizes and discusses the interaction of alphaherpesviruses with actin and cholesterol during different stages of the replication cycle: virus entry, replication and assembly in the nucleus, and virus egress. Elucidating these interactions not only yields novel insights into the biology of these important pathogens, but may also shed new light on cell biological aspects of actin and cholesterol, and lead to novel avenues in the design of antiviral strategies.
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Actinas/metabolismo , Alphaherpesvirinae/fisiología , Colesterol/metabolismo , Infecciones por Herpesviridae/virología , Interacciones Huésped-Patógeno , Animales , Núcleo Celular/virología , Citoesqueleto/virología , Replicación del ADN , Humanos , Ensamble de Virus , Internalización del Virus , Liberación del VirusAsunto(s)
Investigación Biomédica/economía , Investigación Biomédica/organización & administración , Redes Comunitarias/organización & administración , Bélgica , Investigación Biomédica/instrumentación , Tecnología Biomédica/economía , Tecnología Biomédica/instrumentación , Tecnología Biomédica/métodos , Tecnología Biomédica/organización & administración , Movilidad Laboral , Redes Comunitarias/economía , Genómica/economía , Genómica/instrumentación , Genómica/organización & administración , Humanos , Laboratorios/economía , Laboratorios/organización & administración , Personal de Laboratorio/economía , Personal de Laboratorio/organización & administración , Proteómica/economía , Proteómica/instrumentación , Proteómica/organización & administración , EspañaRESUMEN
Viral glycoproteins gB and gD of the swine alphaherpesvirus pseudorabies virus (PRV), which is closely related to human herpes simplex virus and varicella-zoster virus, are able to drive internalization of antibody-antigen complexes that may form at the cell surface of infected monocytes, thereby protecting these cells from efficient antibody-mediated lysis. We found earlier that gB relies on an endocytosis motif in its cytoplasmic domain for its function during this internalization process. Here, we report that the PRV gD protein also contains a functional endocytosis motif (YRLL) in its cytoplasmic domain that drives spontaneous endocytosis of gD from the cell surface early in infection and that acts in concert with the endocytosis motif in gB to contribute to efficient internalization of antibody-antigen complexes in PRV-infected monocytes.
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Endocitosis/fisiología , Herpesvirus Suido 1/inmunología , Herpesvirus Suido 1/patogenicidad , Monocitos/inmunología , Monocitos/virología , Proteínas del Envoltorio Viral/inmunología , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Complejo Antígeno-Anticuerpo/metabolismo , Antígenos Virales/metabolismo , Secuencia de Bases , Línea Celular , ADN Viral/genética , Herpesvirus Suido 1/fisiología , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Señales de Clasificación de Proteína/genética , Señales de Clasificación de Proteína/fisiología , Porcinos , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/fisiologíaRESUMEN
The US3 protein is a viral kinase that is conserved among the Alphaherpesvirinae. Here, we show that US3 of the swine alphaherpesvirus pseudorabies virus causes dramatic alterations in the cytoskeleton, resulting in the formation of long actin- and microtubule-containing cell projections in infected and transfected cells. Analysis with a GFP-labeled virus showed that multiple virus particles move inside the projections toward the tip. GFP-labeled virus could also be found in the cytoplasm of neighboring cells that were in contact with the projections. In addition, projection formation could be inhibited by using the actin-stabilizing drug jasplakinolide and could be induced by using the Rho kinase inhibitor Y27632. Analyzing the effect of these drugs on intercellular virus spread indicated that the observed US3-induced alterations in the host cytoskeleton are associated with enhanced intercellular virus spread, thereby suggesting a previously undescribed aspect of alphaherpesvirus spread.
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Citoesqueleto/ultraestructura , Citoesqueleto/virología , Herpesvirus Suido 1/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Testículo/ultraestructura , Amidas/farmacología , Animales , Cápside/efectos de los fármacos , Cápside/fisiología , Depsipéptidos/farmacología , Herpesvirus Suido 1/efectos de los fármacos , Herpesvirus Suido 1/ultraestructura , Masculino , Microscopía Confocal , Microtúbulos/ultraestructura , Microtúbulos/virología , Piridinas/farmacología , Porcinos , Testículo/virología , Ensayo de Placa ViralRESUMEN
The cytoplasmic domain of pseudorabies virus (PRV) glycoprotein B (gB) contains three putative internalization motifs. Previously, we demonstrated that the tyrosine-based YQRL motif at positions 902 to 905, but not the YMSI motif at positions 864 to 867 or the LL doublet at positions 887 and 888, is required for correct functioning of gB during antibody-mediated internalization of PRV cell surface-bound glycoproteins. In the present study, we demonstrate that the YQRL motif is also crucial to allow spontaneous internalization of PRV gB, and thus, that spontaneous and antibody-mediated internalizations of PRV gB occur through closely related mechanisms. Furthermore, we found that PRV gB colocalizes with the cellular clathrin-associated AP-2 adaptor complex and that this colocalization depends on the YQRL motif. In addition, by coimmunoprecipitation assays, we found that during both spontaneous and antibody-dependent internalization, PRV gB physically interacts with AP-2, and that efficient interaction between gB and AP-2 required an intact YQRL motif. Collectively, these findings demonstrate for the first time that during internalization of an alphaherpesvirus envelope protein, i.e., PRV gB, a specific amino acid sequence in the cytoplasmic tail of the protein interacts with AP-2 and may constitute a common AP-2-mediated mechanism of internalization of alphaherpesvirus envelope proteins.
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Complejo 2 de Proteína Adaptadora/metabolismo , Secuencias de Aminoácidos , Regulación Viral de la Expresión Génica , Herpesvirus Suido 1/patogenicidad , Proteínas del Envoltorio Viral/metabolismo , Subunidades alfa de Complejo de Proteína Adaptadora/metabolismo , Animales , Anticuerpos Antivirales/inmunología , Línea Celular , Endocitosis , Porcinos , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genéticaRESUMEN
Disruption of specific components of the host cytoskeleton has been reported for several viruses and is thought to be beneficial for viral replication and spread. Our previous work demonstrated that infection of swine kidney (SK-6) cells with pseudorabies virus (PRV), a swine alphaherpesvirus, induced actin stress fiber breakdown. In the present study, using several PRV deletion mutants, we found that the US3 serine/threonine (S/T) protein kinase is involved in breakdown of actin stress fibers in different PRV-infected cell lines. Further, by transfection assays, we showed that PRV US3 itself, in the absence of other viral proteins, is able to trigger actin stress fiber breakdown when it is localized in sufficient amounts in the nucleus.
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Actinas/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Línea Celular , Herpesvirus Suido 1 , Hidrólisis , Proteínas ViralesRESUMEN
Pseudorabies virus (PRV), a swine alphaherpesvirus, is capable of causing viremia in vaccinated animals. Two mechanisms that may help PRV avoid recognition by the host immune system during this viremia are direct cell-to-cell spread in tissue and antibody-induced internalization of viral cell surface glycoproteins in PRV-infected blood monocytes, the carrier cells of the virus in the blood. PRV glycoprotein B (gB) is crucial during both processes. Here we show that mutating a tyrosine residue located in a YXXPhi motif in the gB cytoplasmic tail results in decreased efficiency of cell-to-cell spread and a strong reduction in antibody-induced internalization of viral cell surface glycoproteins. Mutating the dileucine motif in the gB tail led to an increased cell-to-cell spread of the virus and the formation of large syncytia.