The Structure of a Biologically Active Influenza Virus Ribonucleoprotein Complex

The influenza viruses contain a segmented, single-stranded RNA genome of negative polarity. Each RNA segment is encapsidated by the nucleoprotein and the polymerase complex into ribonucleoprotein particles (RNPs), which are responsible for virus transcription and replication. Despite their importance, information about the structure of these RNPs is scarce. We have determined the three-dimensional structure of a biologically active recombinant RNP by cryo-electron microscopy. The structure shows a nonameric nucleoprotein ring (at 12 Å resolution) with two monomers connected to the polymerase complex (at 18 Å resolution). Docking the atomic structures of the nucleoprotein and polymerase domains, as well as mutational analyses, has allowed us to define the interactions between the functional elements of the RNP and to propose the location of the viral RNA. Our results provide the first model for a functional negative-stranded RNA virus ribonucleoprotein complex. The structure reported here will serve as a framework to generate a quasi-atomic model of the molecular machine responsible for viral RNA synthesis and to test new models for virus RNA replication and transcription

Ultrastructural and functional analyses of recombinant influenza virus ribonucleoproteins suggest dimerization of nucleoprotein during virus amplification

8 pages, 7 figures.-- PMID: 10590102 [PubMed].-- PMCID: PMC111524.Full-text version available Open Access at PebMed Central: http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=10590102Influenza virus ribonucleoproteins (RNPs) were reconstituted in vivo from cloned cDNAs expressing the three polymerase subunits, the nucleoprotein (NP), and short template RNAs. The structure of purified RNPs was studied by electron microscopy and image processing. Circular and elliptic structures were obtained in which the NP and the polymerase complex could be defined. Comparison of the structure of RNPs of various lengths indicated that each NP monomer interacts with approximately 24 nucleotides. The analysis of the amplification of RNPs with different lengths showed that those with the highest replication efficiency contained an even number of NP monomers, suggesting that the NP is incorporated as dimers into newly synthesized RNPs.J. Ortega was a fellow of Instituto de Estudios Turolenses. This work was supported by Programa Sectorial de Promoción General del Conocimiento (grants PB97-1160 and PB96-0818).Peer reviewe

Monitoring SARS-COV-2 surrogate TGEV individual virions structure survival under harsh physicochemical environments

Effective airborne transmission of coronaviruses via liquid microdroplets requires a virion structure that must withstand harsh environmental conditions. Due to the demanding biosafety requirements for the study of human respiratory viruses, it is important to develop surrogate models to facilitate their investigation. Here we explore the mechanical properties and nanostructure of transmissible gastroenteritis virus (TGEV) virions in liquid milieu and their response to different chemical agents commonly used as biocides. Our data provide two-fold results on virus stability: First, while particles with larger size and lower packing fraction kept their morphology intact after successive mechanical aggressions, smaller viruses with higher packing fraction showed conspicuous evidence of structural damage and content release. Second, monitoring the structure of single TGEV particles in the presence of detergent and alcohol in real time revealed the stages of gradual degradation of the virus structure in situ. These data suggest that detergent is three orders of magnitude more efficient than alcohol in destabilizing TGEV virus particles, paving the way for optimizing hygienic protocols for viruses with similar structure, such as SARS-CoV-

Evidence on port-locking with heparin versus saline in patients with cancer not receiving chemotherapy: A randomized clinical trial

Objective: To assess the safety and efficacy of port-locking with heparin every 2 months vs. every 4 months and vs. saline solution every 2 months in patients with cancer not receiving active chemotherapy. The hypothesis stated that locking with heparin at four-month intervals and saline at two-month intervals would not increment > 10% of port obstructions. Methods: Multicentre, phase IV parallel, post-test control group study took place at the two chemotherapy units of oncology hospitals. Included patients with cancer with ports that completed the chemotherapy treatment but still having port maintenance care or blood samples taken up to four months. A sample of 126 patients with cancer in three arms was needed to detect a maximum difference of 10% for bioequivalence on the locking methods. Consecutive cases non-probabilistic sampling and randomized to one of the three groups; group A: received heparin 60 IU/mL every two months (control) vs. group B heparin every four months and vs. saline every two months in group C. Primary variables were the type of locking regimen, port obstruction, and absence of blood return, port-related infection, or venous thrombosis during the study period. Clinical and sociodemographic variables were also collected. Results: A total of 143 patients were randomly assigned; group A, 47 patients with heparin every 2 months, group B, 51 patients with heparin 4 months, and group C, 45 patients with saline every 2 months. All participants presented an adequate blood return and no obstructions, until the month of the 10th, when one participant in the group A receiving was withdrawn due to an absence of blood flow ( P 1/4 0.587). Conclusions: Port locks with heparin every 4 months or saline every 2 months did not show differences in safety maintenance, infection, or thrombosis compared to heparin every 2 months

Patrones sinópticos aplicados a la predicción en la península antártica

Ponencia presentada en: VI Simposio Nacional de Predicción, celebrado en los servicios centrales de AEMET, en Madrid, del 17 al 19 de septiembre de 2018.Aplicando el método de análisis mediante clústeres al campo de presión en superficie de los reanálisis del modelo ERA Interim, se han definido cinco patrones para el área del paso de Drake y la península antártica. El análisis de frecuencias muestra que los cinco presentan una ocurrencia anual similar pero una gran variabilidad estacional, y la persistencia de cada uno es relativamente alta. La transición entre patrones diferentes tiende a seguir un ciclo acorde con el desplazamiento de un cuarto de onda hacia el este de las ondas sinópticas. El estudio de las configuraciones típicas de la región, basado en los patrones obtenidos, y su relación con el tiempo asociado en superficie (en particular, efemérides) en las bases antárticas españolas (BAE) favorecerá la comprensión de la meteorología de la zona. Además, la aplicación de estos patrones a situaciones cotidianas facilitará y hará más eficiente el trabajo diario de predicción operativa en las campañas antárticas españolas

Fernando de Castro and the discovery of the arterial chemoreceptors

Producción CientíficaWhen de Castro entered the carotid body(CB)field,the organ was considered to be a small autonomic ganglion,a gland,a glomusorglomerulus,or a paraganglion. In his 1928 paper,de Castro concluded:“Insum,the Glomuscarotic umisinnervated by centripetal fibers,whose trophic center sare located in thesensory ganglia of the glossopharyngeal, and not by centrifugal[efferent] or secret o motor fibers a sisthe case for glands ; these are precisely the facts which lead to suppose that the Glomuscaroticumisa sensory organ.”A few pages down,de Castro wrote:“The Glomus represents an organ with multiplereceptors furnished with specialized receptor cells like those of the sensory organs [tastebuds?]...As aplausible hypothesis we propos et hattheGlomuscaroti cum represents a sensory organ, at present the only one in its kind, dedicated to capture certain qualitative variations in the composition of blood, a function that,possibly by are flex mechanism would have an effect on the functionalactivity of other organs... Therefore, thesensory fiber would not be directly stimulated by blood, but via the intermediation of the epithelial cell soft he organ, which, as their structures suggests, possess a secretory function which would participate in the stimulation of the centripetal fibers.”In our article we will recreat et he experiments that allowed Fernando de Castrotoreach this first conclusion. Also, we will scrutinize the natural endowment sand the scientific knowledge that drove de Castrotomaket the triple hypotheses : the CBaschemoreceptor (variationsinbloodcomposition),as a secondary sensory receptor which functioning involves a chemical synapse, and as a center, origin of systemicreflexes. After a brief account of the systemic reflex effects resulting from the CB stimulation, we wil lcomplete our article with a general view of the cellular-molecular mechanisms currently thought to be involved in the functionin go fthis arterial chemoreceptor

Co-responses of bacterial and fungal communities to fire management treatments in Mediterranean pyrophytic ecosystems

Producción CientíficaCistus scrublands are pyrophytic ecosystems and occur widely across Mediterranean regions. Management of these scrublands is critical to prevent major disturbances, such as recurring wildfires. This is because management appears to compromise the synergies necessary for forest health and the provision of ecosystem services. Furthermore, it supports high microbial diversity, opening questions of how forest management practices impact belowground associated diversity as research related to this issue is scarce. This study aims to investigate the effects of different fire prevention treatments and site history on bacterial and fungi co-response and co-occurrence patterns over a fire-risky scrubland ecosystem. Two different site histories were studied by applying three different fire prevention treatments and samples were analyzed by amplification and sequencing of ITS2 and 16S rDNA for fungi and bacteria, respectively. The data revealed that site history, especially regarding fire occurrence, strongly influenced the microbial community. Young burnt areas tended to have a more homogeneous and lower microbial diversity, suggesting environmental filtering to a heat-resistant community. In comparison, young clearing history also showed a significant impact on the fungal community but not on the bacteria. Some bacteria genera were efficient predictors of fungal diversity and richness. For instance, Ktedonobacter and Desertibacter were a predictor of the presence of the edible mycorrhizal bolete Boletus edulis. These results demonstrate fungal and bacterial community co-response to fire prevention treatments and provide new tools for forecasting forest management impacts on microbial communities.Junta de Castilla y León (VA050P17

Aportaciones hidrogeológicas al estudio arqueológico de los orígenes de la Edad del Bronce de La Mancha: la cueva monumentalizada de Castillejo del Bonete (Terrinches, Ciudad Real-España)

Estudios recientes indican que las motillas, asentamientos de la Edad del Bronce de La Mancha, pudieron ser las más antiguas captaciones de agua subterránea en la Península Ibérica. Pero ¿por qué no existen motivas en el Campo de Montiel, territorio ubicado tradicionalmente en esta área cultural? En Castillejo del Bonete, sitio arqueológico situado en esa comarca, existe una cueva que fue utilizada y sellada durante la Prehistoria Reciente. Se presenta ahora la primera investigación paleohidrogeológica interdisciplinar en La Mancha, que ha analizado manantiales y niveles de agua subterránea del acuífero de Campo de Montiel, así como el interior de la sima de Castillejo del Bonete. Las conclusiones avanzan una relación entre el sustrato hidrogeomorfológico y la distribución espacial de las motillas. Además permiten descartar que la sima de Castillejo del Bonete fuera utilizada como mina o como acceso al acuífero, dos de las hipótesis de trabajo planteadas. De ese modo cobra fuerza que Castillejo del Bonete fuera un excepcional monumento funerario y simbólico durante el Calcolítico y la Edad del Bronce, perteneciente a una nueva clase de asentamientos desconocidos hasta ahora en el grupo cultural de la Edad del Bronce de La Mancha

Measurement of galactosyl-deficient IgA1 by the monoclonal antibody KM55 contributes to predicting patients with IgA nephropathy with high risk of long-term progression

Backgroundandobjective:About25%ofpatientswithIgAnephropathy(IgAN)progresstostage5chronickidneydisease(CKD)afteryearsofevolution.Varioustoolshavebeendevelopedinrecentyearsdesignedtopredictwhichofthepatientswillhadpooreroutcomes.Thevalueofcirculatinggalactosyl-deficientIgA1(Gd-IgA1)hasbeenrelatedtoaworseevolutionofIgANinseveralstudies.TherearealsosomepublicationsthatrelatehigherAPRILvalueswithaworseevolution.Recently,anewmethodhasbeendevelopedthatallowsmeasuringthevalueofcirculatingGd-IgA1inasimplerwaythanthosepreviouslyavailable.TheobjectiveofthisstudyistoanalyzetheinfluenceofcirculatingGd-IgA1,measuredbythismethod,ontheprogressionofIgAN.Materialsandmethods:Forty-ninepatientswithadiagnosisofIgANdemonstratedbyrenalbiopsywereselectedinourcenter,withouthavingreceivedpriorimmunosuppressivetreat-ment,forwhomfrozenserumwasavailable.Themedianfollow-upwas4years.Gd-IgA1wasmeasuredbylectin-independentELISAwiththemonoclonalantibodyKM55(IgA1kitCat.No.30111694.IBLInt.,Hamburg,Germany).Likewise,APRILlevelswerealsomeasuredinthesepatients.Results:19(38.8%)patientsreachedstage5CKD.ThefourthquartileofcirculatingGd-IgA1wasrelatedtoahighercumulativeriskofreachingstage5CKDintheKaplan?Meieranalysis(riskatthe5thyear39.4%vs.24.3%,logrankp=0.019).TheGd-IgA1valuewasrelatedto anincreasedriskofCKDstage5(HR1.147,95%CI1.035?1.270,p=0.009),regardlessofglomerularfiltrationrate,proteinuria,thepercentageofsclerosedglomeruliandthevalueofsegmentalsclerosis.WedidnotfindsignificantdifferencesintheAPRILvalues.Conclusions:ThevalueofcirculatingGd-IgA1measuredbythemonoclonalantibodyKM55isrelatedtoaworseevolutionofpatientswithIgANindependentlyofothervariables,soitcouldbeincludedinthestudyofpatientstoimprovethepredictionoftheriskofdiseaseprogression

Simulation of electron transport and secondary emission in a photomultiplier tube and experimental validation

[EN] The electron amplification and transport within a photomultiplier tube (PMT) has been investigated by developing an in-house Monte Carlo simulation code. The secondary electron emission in the dynodes is implemented via an effective electron model and the Modified Vaughan¿s model, whereas the transport is computed with the Boris leapfrog algorithm. The PMT gain, rise time and transit time have been studied as a function of supply voltage and external magnetostatic field. A good agreement with experimental measurements using a Hamamatsu R13408-100 PMT was obtained. The simulations have been conducted following different treatments of the underlying geometry: three-dimensional, two-dimensional and intermediate (2.5D). The validity of these approaches is compared. The developed framework will help in understanding the behavior of PMTs under highly intense and irregular illumination or varying external magnetic fields, as in the case of prompt gamma-ray measurements during pencil-beam proton therapy; and aid in optimizing the design of voltage dividers with behavioral circuit models.This work was supported by Conselleria de Educación, Investigación, Cultura y Deporte (Generalitat Valenciana) under grant numbers CDEIGENT/2019/011 and CDEIGENT/2021/012. P. Martín-Luna is supported by the Ministerio de Universidades (Gobierno de España), Spain under Grant Number FPU20/04958. We thank Hamamatsu (V. Sánchez, D. Castrillo) for technical support and guidance; R. Carrasco (IFIC) and P. Wohlfahrt (Siemens Healthineers) for the CT scanning; D. Calvo and D. Real (KM3net-IFIC) for their LED test platform, the electronics and maintenance services at IFIC for excellent support; and K. Albiol, J. V. Casaña-Copado, A. Gallas Torreira, E. Lemos Cid, G. Pausch, A. Pazos Álvarez, E. Pérez Trigo, S. Rit, A. Ros, J. Roser, J. Stein, J. L. Taín and R. Viegas for useful discussions.Martín-Luna, P.; Esperante, D.; Fernández Prieto, A.; Fuster-Martínez, N.; García Rivas, I.; Gimeno, B.; Ginestar Peiro, D.... (2024). Simulation of electron transport and secondary emission in a photomultiplier tube and experimental validation. Sensors and Actuators A Physical. 365:1-10. https://doi.org/10.1016/j.sna.2023.11485911036