.

Benign Prostatic Hyperplasia (BPH) is the result of excessive cellullar proliferation in the prostatic transition zone, that compresses the urethra, causing the symptoms of the disease. Evidence supports that atherogenic environment could contribute to the development and progression of BPH, increasing its incidence and aggressiveness. The BPH hyperproliferative state requires coordinated communication between the different components that maintain a permissive microenvironment. Thus, extracellular vesicles (EVs) have become relevant as intercellular communication regulators in multiple processes. It has been described that oxidized-LDL molecule (OxLDL) would be involved in numerous signaling pathways; including signaling by EVs promoting cellullar proliferation. EVs, may contain diverse biomolecules that confer them functions in cellular communication. In addition, it has been reported that EVs derived from different cell types are capable to mediate proliferative and inflammatory effects. In this context, we proposed as objectives: 1) to evaluate the effect of OxLDL, simulating an atherogenic state, on cell proliferation in primary cultures of human prostate stromal cells (HPSC) from patient samples (n=8) from the Sanatorio Allende of Córdoba; 2) to isolate EVs from primary cultures by differential ultracentrifugation and analyze their production and release into the medium in treatments with OxLDL vs. vehicle; 3) to morphologically characterize EVs by transmission electron microscopy (TEM) and confirm the identity and presence of exosomes, through CD63 immuno-staining using colloidal gold. It was observed that OxLDL (20μM) produced a significant increase in cell proliferation rate compared to vehicle. EVs were obtained by differential ultracentrifugations (2k, 10k, 150k pellets) and visualized by TEM using negative staining. HPSC from patients with BPH showed a very low frequency of EVs released, with OxLDL inducing a 10-fold increase, especially in the 15-20nm fraction (p<0.001). Ultrastructurally, these EVs exhibited a spherical and concave appearance, compatible with exosomes. Therefore, they were positively verified by CD63 immunostaining. Finally, we conclude that OxLDL would favor cell proliferation, increase and release of EVs, which would participate in cell communication and maintenance of the permissive environment, propitious to progression and increased aggressiveness of BPH.La Hiperplasia Prostática Benigna (HPB) surge a consecuencia de la proliferación celular excesiva de la zona de transición prostática que comprime la uretra, provocando la sintomatología propia de la enfermedad. Evidencias sostienen que el entorno aterogénico podrían contribuir al desarrollo y progresión de la HPB, aumentando su incidencia y agresividad. El estado hiperproliferativo de la HPB requiere una coordinada comunicación entre los diferentes componentes que mantienen un microambiente permisivo. Así, las vesículas extracelulares (EVs) han tomado relevancia como reguladores de comunicación intercelular en múltiples procesos. Se ha descrito que la molécula de LDL-oxidada (OxLDL) estaría involucrada en diversas vías de señalización; incluida la señalización por EVs promoviendo la proliferación celular. Las EVs pueden contener diversas biomoléculas que les confieren funciones en comunicación intercelular y, además, se reportaron efectos proliferativos e inflamatorios mediados por EVs derivadas de distintos tipos celulares. Es este contexto, propusimos como objetivos: 1) evaluar el efecto de OxLDL, simulando un estado aterogénico, sobre la proliferación celular en cultivos primarios de células estromales prostáticas humanas (HPSC) de muestras de pacientes (n=8) del Sanatorio Allende de Córdoba; 2) aislar EVs de cultivos primarios mediante ultracentrifugaciones diferenciales y analizar su producción y liberación al medio en tratamientos con OxLDL vs. vehículo; 3) caracterizar morfológicamente EVs mediante microscopía electrónica de transmisión (TEM) y confirmar la identidad y presencia de exosomas, a través de inmuno-marcación de CD63 utilizando oro coloidal. Se observó que OxLDL (20μM) produjo un aumento significativo en la tasa de proliferación celular respecto al vehículo. EVs fueron obtenidas mediante utracentrifugaciones diferenciales (pellets 2k, 10k, 150k) y visualizadas por TEM utilizando tinción negativa. Las HPSC de pacientes con HPB mostraron una frecuencia muy baja de EVs liberadas, con OxLDL induciendo un aumento de 10 veces, especialmente en la fracción 15-20nm (p<0,001). Ultraestructuralmente, estas EVs exhibieron una apariencia esférica y cóncava, compatible con exosomas; luego fueron verificados positivamente por inmuno-marcación de CD63. Finalmente, podemos concluir que OxLDL favorecería la proliferación celular, incremento y liberación de EVs, las cuales participarían en comunicación celular y mantención del ambiente permisivo propicio para la progresión y aumento de agresividad de la HPB.

The CARMENES search for exoplanets around M dwarfs: Two planets on opposite sides of the radius gap transiting the nearby M dwarf LTT 3780

Full list of authors: Nowak, G.; Luque, R.; Parviainen, H.; Pallé, E.; Molaverdikhani, K.; Béjar, V. J. S.; Lillo-Box, J.; Rodríguez-López, C.; Caballero, J. A.; Zechmeister, M.; Passegger, V. M.; Cifuentes, C.; Schweitzer, A.; Narita, N.; Cale, B.; Espinoza, N.; Murgas, F.; Hidalgo, D.; Zapatero Osorio, M. R.; Pozuelos, F. J.; Aceituno, F. J.; Amado, P. J.; Barkaoui, K.; Barrado, D.; Bauer, F. F.; Benkhaldoun, Z.; Caldwell, D. A.; Casasayas Barris, N.; Chaturvedi, P.; Chen, G.; Collins, K. A.; Collins, K. I.; Cortés-Contreras, M.; Crossfield, I. J. M.; de León, J. P.; Díez Alonso, E.; Dreizler, S.; El Mufti, M.; Esparza-Borges, E.; Essack, Z.; Fukui, A.; Gaidos, E.; Gillon, M.; Gonzales, E. J.; Guerra, P.; Hatzes, A.; Henning, Th.; Herrero, E.; Hesse, K.; Hirano, T.; Howell, S. B.; Jeffers, S. V.; Jehin, E.; Jenkins, J. M.; Kaminski, A.; Kemmer, J.; Kielkopf, J. F.; Kossakowski, D.; Kotani, T.; Kürster, M.; Lafarga, M.; Latham, D. W.; Law, N.; Lissauer, J. J.; Lodieu, N.; Madrigal-Aguado, A.; Mann, A. W.; Massey, B.; Matson, R. A.; Matthews, E.; Montañés-Rodríguez, P.; Montes, D.; Morales, J. C.; Mori, M.; Nagel, E.; Oshagh, M.; Pedraz, S.; Plavchan, P.; Pollacco, D.; Quirrenbach, A.; Reffert, S.; Reiners, A.; Ribas, I.; Ricker, G. R.; Rose, M. E.; Schlecker, M.; Schlieder, J. E.; Seager, S.; Stangret, M.; Stock, S.; Tamura, M.; Tanner, A.; Teske, J.; Trifonov, T.; Twicken, J. D.; Vanderspek, R.; Watanabe, D.; Wittrock, J.; Ziegler, C.; Zohrabi, F.We present the discovery and characterisation of two transiting planets observed by the Transiting Exoplanet Survey Satellite (TESS) orbiting the nearby (d∗ ≈ 22 pc), bright (J ≈ 9 mag) M3.5 dwarf LTT 3780 (TOI-732). We confirm both planets and their association with LTT 3780 via ground-based photometry and determine their masses using precise radial velocities measured with the CARMENES spectrograph. Precise stellar parameters determined from CARMENES high-resolution spectra confirm that LTT 3780 is a mid-M dwarf with an effective temperature of Teff = 3360 ± 51 K, a surface gravity of log g∗ = 4.81 ± 0.04 (cgs), and an iron abundance of [Fe/H] = 0.09 ± 0.16 dex, with an inferred mass of M∗ = 0.379 ± 0.016M· and a radius of R∗ = 0.382 ± 0.012R·. The ultra-short-period planet LTT 3780 b (Pb = 0.77 d) with a radius of 1.35-0.06+0.06 R·, a mass of 2.34-0.23+0.24 M·, and a bulk density of 5.24-0.81+0.94 g cm-3 joins the population of Earth-size planets with rocky, terrestrial composition. The outer planet, LTT 3780 c, with an orbital period of 12.25 d, radius of 2.42-0.10+0.10 R·, mass of 6.29-0.61+0.63 M·, and mean density of 2.45-0.37+0.44 g cm-3 belongs to the population of dense sub-Neptunes. With the two planets located on opposite sides of the radius gap, this planetary system is anexcellent target for testing planetary formation, evolution, and atmospheric models. In particular, LTT 3780 c is an ideal object for atmospheric studies with the James Webb Space Telescope (JWST). © 2020 ESO.CARMENES is an instrument for the Centro Astronomico Hispano-Aleman de Calar Alto (CAHA, Almeria, Spain). CARMENES is funded by the German Max-Planck-Gesellschaft (MPG), the Spanish Consejo Superior de Investigaciones Cientificas (CSIC), the European Union through FEDER/ERF FICTS-2011-02 funds, and the members of the CARMENES Consortium (Max-Planck-Institut fur Astronomie, Instituto de Astrofisica de Andalucia, Landessternwarte Konigstuhl, Institut de Ciencies de l'Espai, Institut fur Astrophysik Gottingen, Universidad Complutense de Madrid, Thuringer Landessternwarte Tautenburg, Instituto de Astrofisica de Canarias, Hamburger Sternwarte, Centro de Astrobiologia and Centro Astronomico Hispano-Aleman), with additional contributions by the Spanish Ministry of Economy, the German Science Foundation through the Major Research Instrumentation Programme and DFG Research Unit FOR2544 "Blue Planets around Red Stars", the Klaus Tschira Stiftung, the states of Baden-Wurttemberg and Niedersachsen, and by the Junta de Andalucia. This paper includes data collected by the TESS mission. Funding for the TESS mission is provided by the NASA Explorer Program. We acknowledge the use of public TOI Release data from pipelines at the TESS Science Office and at the TESS Science Processing Operations Center. Resources supporting this work were provided by the NASA High-End Computing (HEC) Program through the NASA Advanced Supercomputing (NAS) Division at Ames Research Center for the production of the SPOC data products. This research has made use of the Exoplanet Follow-up Observation Program website, which is operated by the California Institute of Technology, under contract with the National Aeronautics and Space Administration under the Exoplanet Exploration Program. This work has made use of data from the European Space Agency (ESA) mission Gaia (https://www.cosmos.esa.int/gaia), processed by the Gaia Data Processing and Analysis Consortium (DPAC, https://www.cosmos.esa.int/web/gaia/dpac/consortium).Funding for the DPAC has been provided by national institutions, in particular the institutions participating in the Gaia Multilateral Agreement. This article is partly based on observations made with the MuSCAT2 instrument, developed by ABC, at Telescopio Carlos Sanchez operated on the island of Tenerife by the IAC in the Spanish Observatorio del Teide. This work makes use of observations from the LCOGT network. This work makes use of observations acquired with the T150 telescope at Sierra Nevada Observatory, operated by the Instituto de Astrofisica de Andalucia (IAACSIC). Some of the Observations in the paper made use of the High-Resolution Imaging instrument 'Alopeke at Gemini-North. `Alopeke was funded by the NASA Exoplanet Exploration Program and built at the NASA Ames Research Center by Steve B. Howell, Nic Scott, Elliott P. Horch, and Emmett Quigley. IRD is operated by the Astrobiology Center of the National Institutes of Natural Sciences. The research leading to these results has received funding from the ARC grant for Concerted Research Actions, financed by the WalloniaBrussels Federation. TRAPPIST is funded by the Belgian Fund for Scientific Research (Fond National de la Recherche Scientifique, FNRS) under the grant FRFC 2.5.594.09.F, with the participation of the Swiss National Science Fundation (SNF). TRAPPIST-North is a project funded by the University of Liege (Belgium), in collaboration with Cadi Ayyad University of Marrakech (Morocco) M.G. and E.J. are F.R.S.-FNRS Senior Research Associate. The authors acknowledge funding from the Spanish Ministry of Economics and Competitiveness through projects PGC2018-098153-B-C31 and AYA2015-69350-C3-2-P. This work is partly supported by JSPS KAKENHI Grant Numbers JP18H01265 and JP18H05439, and JST PRESTO Grant Number JPMJPR1775. V.M.P. acknowledges support from NASA Grant NNX17AG24G. T.H. acknowledges support from the European Research Council under the Horizon 2020 Framework Program via the ERC Advanced Grant Origins 83 24 28. This research has been partially funded by Project No. MDM-2017-0737 Unidad de Excelencia "Maria de Maeztu" -Centro de Astrobiologia (INTA-CSIC). This research acknowledges financial support from the State Agency for Research of the Spanish MCIU through the "Center of Excellence Severo Ochoa" award to the Instituto de Astrofisica de Andalucia (SEV-2017-0709) and project AYA2016-794425

Intracellular interferons in fish : a unique means to combat viral infection

Peer reviewedPublisher PD

Transcription of toll-like receptors 2, 3, 4 and 9, FoxP3 and Th17 cytokines in a susceptible experimental model of canine Leishmania infantum infection

Canine leishmaniosis (CanL) due to Leishmania infantum is a chronic zoonotic systemic disease resulting from complex interactions between protozoa and the canine immune system. Toll-like receptors (TLRs) are essential components of the innate immune system and facilitate the early detection of many infections. However, the role of TLRs in CanL remains unknown and information describing TLR transcription during infection is extremely scarce. The aim of this research project was to investigate the impact of L. infantum infection on canine TLR transcription using a susceptible model. The objectives of this study were to evaluate transcription of TLRs 2, 3, 4 and 9 by means of quantitative reverse transcription polymerase chain reaction (qRT-PCR) in skin, spleen, lymph node and liver in the presence or absence of experimental L. infantum infection in Beagle dogs. These findings were compared with clinical and serological data, parasite densities in infected tissues and transcription of IL-17, IL-22 and FoxP3 in different tissues in non-infected dogs (n = 10), and at six months (n = 24) and 15 months (n = 7) post infection. Results revealed significant down regulation of transcription with disease progression in lymph node samples for TLR3, TLR4, TLR9, IL-17, IL-22 and FoxP3. In spleen samples, significant down regulation of transcription was seen in TLR4 and IL-22 when both infected groups were compared with controls. In liver samples, down regulation of transcription was evident with disease progression for IL-22. In the skin, upregulation was seen only for TLR9 and FoxP3 in the early stages of infection. Subtle changes or down regulation in TLR transcription, Th17 cytokines and FoxP3 are indicative of the silent establishment of infection that Leishmania is renowned for. These observations provide new insights about TLR transcription, Th17 cytokines and Foxp3 in the liver, spleen, lymph node and skin in CanL and highlight possible markers of disease susceptibility in this model

A systematic review of the evidence for single stage and two stage revision of infected knee replacement

BACKGROUND: Periprosthetic infection about the knee is a devastating complication that may affect between 1% and 5% of knee replacement. With over 79 000 knee replacements being implanted each year in the UK, periprosthetic infection (PJI) is set to become an important burden of disease and cost to the healthcare economy. One of the important controversies in treatment of PJI is whether a single stage revision operation is superior to a two-stage procedure. This study sought to systematically evaluate the published evidence to determine which technique had lowest reinfection rates. METHODS: A systematic review of the literature was undertaken using the MEDLINE and EMBASE databases with the aim to identify existing studies that present the outcomes of each surgical technique. Reinfection rate was the primary outcome measure. Studies of specific subsets of patients such as resistant organisms were excluded. RESULTS: 63 studies were identified that met the inclusion criteria. The majority of which (58) were reports of two-stage revision. Reinfection rated varied between 0% and 41% in two-stage studies, and 0% and 11% in single stage studies. No clinical trials were identified and the majority of studies were observational studies. CONCLUSIONS: Evidence for both one-stage and two-stage revision is largely of low quality. The evidence basis for two-stage revision is significantly larger, and further work into direct comparison between the two techniques should be undertaken as a priority

Green-to-red photoconvertible fluorescent proteins: tracking cell and protein dynamics on standard wide-field mercury arc-based microscopes

<p>Abstract</p> <p>Background</p> <p>Green fluorescent protein (GFP) and other FP fusions have been extensively utilized to track protein dynamics in living cells. Recently, development of photoactivatable, photoswitchable and photoconvertible fluorescent proteins (PAFPs) has made it possible to investigate the fate of discrete subpopulations of tagged proteins. Initial limitations to their use (due to their tetrameric nature) were overcome when monomeric variants, such as Dendra, mEos, and mKikGR were cloned/engineered.</p> <p>Results</p> <p>Here, we report that by closing the field diaphragm, selective, precise and irreversible green-to-red photoconversion (330-380 nm illumination) of discrete subcellular protein pools was achieved on a wide-field fluorescence microscope equipped with standard DAPI, Fluorescein, and Rhodamine filter sets and mercury arc illumination within 5-10 seconds. Use of a DAPI-filter cube with long-pass emission filter (LP420) allowed the observation and control of the photoconversion process in real time. Following photoconversion, living cells were imaged for up to 5 hours often without detectable phototoxicity or photobleaching.</p> <p>Conclusions</p> <p>We demonstrate the practicability of this technique using Dendra2 and mEos2 as monomeric, photoconvertible PAFP representatives fused to proteins with low (histone H2B), medium (gap junction channel protein connexin 43), and high (α-tubulin; clathrin light chain) dynamic cellular mobility as examples. Comparable efficient, irreversible green-to-red photoconversion of selected portions of cell nuclei, gap junctions, microtubules and clathrin-coated vesicles was achieved. Tracking over time allowed elucidation of the dynamic live-cycle of these subcellular structures. The advantage of this technique is that it can be performed on a standard, relatively inexpensive wide-field fluorescence microscope with mercury arc illumination. Together with previously described laser scanning confocal microscope-based photoconversion methods, this technique promises to further increase the general usability of photoconvertible PAFPs to track the dynamic movement of cells and proteins over time.</p

Pyrosequencing of Bacterial Symbionts within Axinella corrugata Sponges: Diversity and Seasonal Variability

Background: Marine sponge species are of significant interest to many scientific fields including marine ecology, conservation biology, genetics, host-microbe symbiosis and pharmacology. One of the most intriguing aspects of the sponge ‘‘holobiont’’ system is the unique physiology, interaction with microbes from the marine environment and the development of a complex commensal microbial community. However, intraspecific variability and temporal stability of sponge-associated bacterial symbionts remain relatively unknown. Methodology/Principal Findings: We have characterized the bacterial symbiont community biodiversity of seven different individuals of the Caribbean reef sponge Axinella corrugata, from two different Florida reef locations during variable seasons using multiplex 454 pyrosequencing of 16 S rRNA amplicons. Over 265,512 high-quality 16 S rRNA sequences were generated and analyzed. Utilizing versatile bioinformatics methods and analytical software such as the QIIME and CloVR packages, we have identified 9,444 distinct bacterial operational taxonomic units (OTUs). Approximately 65,550 rRNA sequences (24%) could not be matched to bacteria at the class level, and may therefore represent novel taxa. Differentially abundant classes between seasonal Axinella communities included Gammaproteobacteria, Flavobacteria, Alphaproteobacteria, Cyanobacteria, Acidobacter and Nitrospira. Comparisons with a proximal outgroup sponge species (Amphimedon compressa), and the growing sponge symbiont literature, indicate that this study has identified approximately 330 A. corrugata-specific symbiotic OTUs, many of which are related to the sulfur-oxidizing Ectothiorhodospiraceae. This family appeared exclusively within A. corrugata, comprising \u3e34.5% of all sequenced amplicons. Other A. corrugata symbionts such as Deltaproteobacteria, Bdellovibrio, and Thiocystis among many others are described. Conclusions/Significance: Slight shifts in several bacterial taxa were observed between communities sampled during spring and fall seasons. New 16 S rDNA sequences and concomitant identifications greatly expand the microbial community profile for this model reef sponge, and will likely be useful as a baseline for any future comparisons regarding sponge microbial community dynamics

Search for CP violation in D+→ϕπ+ and D+s→K0Sπ+ decays

A search for CP violation in D + → ϕπ + decays is performed using data collected in 2011 by the LHCb experiment corresponding to an integrated luminosity of 1.0 fb−1 at a centre of mass energy of 7 TeV. The CP -violating asymmetry is measured to be (−0.04 ± 0.14 ± 0.14)% for candidates with K − K + mass within 20 MeV/c 2 of the ϕ meson mass. A search for a CP -violating asymmetry that varies across the ϕ mass region of the D + → K − K + π + Dalitz plot is also performed, and no evidence for CP violation is found. In addition, the CP asymmetry in the D+s→K0Sπ+ decay is measured to be (0.61 ± 0.83 ± 0.14)%