IL-10 deficiency exacerbates the brain inflammatory response to permanent ischemia without preventing resolution of the lesion

El pdf del artículo es la versión post-print.Stroke induces inflammation that can aggravate brain damage. This work examines whether interleukin-10 (IL-10) deficiency exacerbates inflammation and worsens the outcome of permanent middle cerebral artery occlusion (pMCAO). Expression of IL-10 and IL-10 receptor (IL-10R) increased after ischemia. From day 4, reactive astrocytes showed strong IL-10R immunoreactivity. Interleukin-10 knockout (IL-10 KO) mice kept in conventional housing showed more mortality after pMCAO than the wild type (WT). This effect was associated with the presence of signs of colitis in the IL-10 KO mice, suggesting that ongoing systemic inflammation was a confounding factor. In a pathogen-free environment, IL-10 deficiency slightly increased infarct volume and neurologic deficits. Induction of proinflammatory molecules in the IL-10 KO brain was similar to that in the WT 6 hours after ischemia, but was higher at day 4, while differences decreased at day 7. Deficiency of IL-10 promoted the presence of more mature phagocytic cells in the ischemic tissue, and enhanced the expression of M2 markers and the T-cell inhibitory molecule CTLA-4. These findings agree with a role of IL-10 in attenuating local inflammatory reactions, but do not support an essential function of IL-10 in lesion resolution. Upregulation of alternative immunosuppressive molecules after brain ischemia can compensate, at least in part, the absence of IL-10. © 2013 ISCBFM.Work supported by the Spanish Ministry of Economy (SAF2011-30492), and the European Community (FP7, grant agreements: n°201024 ARISE and n°278850 InMiND), and the ERANET-NEURON project (PRI-PIMNEU-2011-1342). IPP and EBT had PhD fellowships from the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) of the Generalitat de Catalunya and the FPU program of the Spanish Ministry of Economy, respectively.Peer Reviewe

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg$^{-1}$ for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH$_{2}$-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg$^{-1}$. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g$^{-1}$ and specific activities of up to 29.9 ± 0.3 U mg$^{-1}$, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules

Unexpected discovery: “A new 3,3′-bipyrazolo[3,4-b]pyridine scaffold and its comprehensive analysis”

In this study, a novel 3,3′-bipyrazolo [3,4-b]pyridine-type structure was synthesized from 5-acetylamino-3-methyl-1-phenylpyrazole using the Vilsmeier-Haack reaction as a key step. The spectroscopic properties and structural elucidation of the compound were determined with the use of FT-IR, HRMS, 1H NMR, and 13C NMR. Likewise, the theoretical analysis of the IR and NMR spectra allowed peaks to be assigned and a solid correlation was demonstrated between the experimental and theoretical results. Finally, ab initio calculations based on the density functional theory method at the B3LYP/6-311G (d,p) level of theory were used to determine the conformational energy barrier, facilitating the identification of the most probable conformers of the synthesized compound. Overall, our findings contribute to the understanding of bipyrazolo [3,4-b]pyridine derivatives.Fondo Nacional de Desarrollo Científico y Tecnológico (Chile)9 página

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg−1 for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH2-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg−1. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g−1 and specific activities of up to 29.9 ± 0.3 U mg−1, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules.This project received funding from the European Union’s Horizon 2020 research and innovation program Blue Growth: Unlockingthe potential of Seas and Oceans under grant agreement no. 634486 (project acronym INMARE). This research was also supported by the grants PCIN-2014-107 (within ERA NET IB2 grant nr. ERA-IB-14-030 - MetaCat), PCIN-2017-078 (within the ERA-MarineBiotech grant ProBone), BIO2014-54494-R, MAT2016-77496-R, BIO2017-85522-R, and CTQ2016-79138-R from the Spanish Ministry of Economy, Industry and Competitiveness. A.B. acknowledges the support from the Spanish Ministry of Economy, Industry and Competitiveness (MAT2017-88808-R grant), María de Maeztu Units of Excellence Programme (MDM-2016-0618), and the Diputación de Guipúzcoa for current funding in the frame of Gipuzkoa Fellows program. G.D. thanks the German Federal Ministry of Education and Research (BMBF, Grant No. 031A095C) for funding in the frame of the Molecular Interaction Engineering program (Biotechnologie 2020+). The authors gratefully acknowledge financial support provided by the European Regional Development Fund (ERDF). C.C. thanks the Spanish Ministry of Economy, Industry and Competitiveness for a PhD fellowship (Grant BES-2015-073829).Peer ReviewedPostprint (published version

Influence of different bottom types on the Senegal sole Solea senegalensis Kaup, 1858 ongrowing

Fish coloration may affect its market value. The present paper compares growth, survival rate and pigmentation during the ongrowing of Senegal sole Solea senegalensis Kaup, 1858 on three different bottoms. This experiment lasted five months, and at the beginning fish were 4.93 ± 0.75 cm (length) and 1.91 ± 0.68 g (weight). No significant differences were found for final length and weight between different bottoms. Several pigmentation patterns were found on the sand and white bottoms, but one single pattern appeared in the dark bottom tanks. The survival rate on the sand bottom was significantly less than others, probably due to pathologies stemming from the difficult maintenance of the sand bed.La falta de homogeneidad en la coloración de los peces puede afectar a su valor comercial. En este trabajo se ha comparado el crecimiento, la supervivencia y la pigmentación de ejemplares de lenguado senegalés Solea senegalensis Kaup, 1858 durante el preengorde en tanques con tres tipos de fondos. El experimento tuvo una duración de 5 meses. El tamaño inicial de los individuos era 4,93 ± 0,75 cm y 1,91 ± 0,68 g, y al final de la experiencia no se encontraron diferencias significativas entre las tallas y los pesos de los ejemplares de los distintos tanques. Se detectaron varios patrones de pigmentación en los especímenes correspondientes a los fondos arenoso y blanco, pero solo uno en los de los tanques de fondo gris. La supervivencia en el fondo de arena fue la menor de todas (p 0,01), probablemente atribuible a patologías asociadas al difícil mantenimiento del lecho arenoso.Instituto Español de Oceanografí

White dwarf-main-sequence binaries from Gaia EDR3: The unresolved 100 pc volume-limited sample

We use the data provided by the Gaia Early Data Release 3 to search for a highly-complete volume-limited sample of unresolved binaries consisting of a white dwarf and a main sequence companion (i.e. WDMS binaries) within 100 pc. We select 112 objects based on their location within the Hertzsprung-Russell diagram, of which 97 are new identifications. We fit their spectral energy distributions (SED) with a two-body fitting algorithm implemented in VOSA (Virtual Observatory SED Analyser) to derive the effective temperatures, luminosities and radii (hence surface gravities and masses) of both componentsARM acknowledges financial support from the MINECO under the Ramón y Cajal program (RYC-2016-20254). ST and ARM acknowledge support from the MINECO under the AYA2017-86274-P grant, and the AGAUR grant SGR-661/2017. ESM and FJE acknowledge financial support from the MINECO under the AYA2017-86274-P grant. FJE acknowledges support from the H2020 ESCAPE project (Grant Agreement no. 824064). LMC, LGA and AHC acknowledge support from AGENCIA through the Programa de Modernización Tecnológica BID 1728/OC-AR, and from CONICET through the PIP 2017-2019 GI grant. This publication makes use of VOSA and SVO DiscTool, developed under the Spanish Virtual Observatory project supported from the Spanish MINECO through grant AyA2017-84089. This research has made use of Aladin sky atlas developed at CDS, Strasbourg Observatory, France (Bonnarel et al. 2000; Boch & Fernique 2014). TOPCAT (Taylor 2005) and STILTS (Taylor 2006) have also been widely used in this paper. We thank the anonymous referee for the helpful suggestions. The authors are greatly indebted to Detlev Koester for sharing his grid of model atmosphere white dwarf spectra. The authors also thank Roberto Raddi for sharing the grid of white dwarf absolute magnitudes calculated for the Gaia EDR3 bandpasses. 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 AgreementPostprint (updated version

Analysis and description of disease-specific quality of life in patients with anal fistula

INTRODUCTION: In patients diagnosed with anal fistula, knowing the quality of life specifically related to the disease can help coloproctology specialists to choose the most appropriate therapeutic strategy for each case. The aim of our study is to analyzse and describe the factors related to the specific quality of life in a consecutive series of patients diagnosed with anal fistula. METHODS: Observational, cross-sectional study carried out from March 2015 to February 2017. All patients were assessed in the colorectal surgery unit of a hospital in southeast of Spain. After performing an initial anamnesis and a physical examination, patients diagnosed with anal fistula completed the Quality of Life in Ppatients with Anal Fistula Questionnaire (QoLAF-Q). This questionnaire specifically measures quality of life in people with anal fistula and its score range is the following: zero impact = 14 points, limited impact = 15 to 28 points, moderate impact = 29 to 42 points, high impact = 43 to 56 points, and very high impact = 57 to 70 points. RESULTS: A total of 80 patients were included. The median score obtained in the questionnaire for the sample studied was 34.00 (range=14-68). Statistically significant differences between patients with "primary anal fistula" (n=65) and "recurrent anal fistula" (n=15) were observed (mean rank=42.96 vs. mean rank=29.83, p=0.048). Furthermore, an inverse proportion (P=.016) between "time with clinical symptoms" and "impact on quality of life" was found (5 years: mean rank = 19.00). There were no statistically significant differences (P=.149) between quality of life amongst patients diagnosed with complex (mean rank = 36.13) and simple fistulae (mean rank = 43.59). CONCLUSIONS: Anal fistulae exert moderate-high impact on patients' quality of life. "Shorter time experiencing clinical symptoms" and the "presence of primary fistula" are factors that can be associated with worse quality of life

The effect of diet and sociopolitical change on physiological stress and behavior in late Roman‐Early Byzantine (300–700 AD) and Islamic (902–1,235 AD) populations from Ibiza, Spain

Objectives This study evaluated chronological changes in physiological stress and levels of habitual loading of Ibizan populations from the Late Roman-Early Byzantine to the Islamic period (300-1235 AD) using measures of body size and bone cross-sectional properties. It also explored the effect of diet, modeled using stable isotopes, on physiological stress levels and behavior. Materials and Methods American Journal of Physical Anthropology, Volume 172, Issue 2, June 2020 pp. 189-213 DOI:10.1002/ajpa.24062 Published by Wiley. This is the Author Accepted Manuscript issued with: Creative Commons Attribution Non-Commercial License (CC:BY:NC 4.0). The final published version (version of record) is available online at DOI:10.1002/ajpa.24062. Please refer to any applicable publisher terms of use. 2 The sample comprised individuals from three archaeological populations: Urban Late Roman- Early Byzantine (LREB) (300-700 AD), Medieval Urban Islamic (902-1235 AD), and Rural Islamic. Bone lengths, femoral head dimensions, and diaphyseal products and circumferences were compared to assess differences in body size and habitual loading in 222 adult individuals. Ordinary least squares regression evaluated the correlations between these measures and carbon (δ13C) and nitrogen (δ15N) stable isotope ratios in 115 individuals for whom both isotope values and osteological measures are available. Results The Rural Islamic group had shorter stature and reduced lower limb cross-sectional properties compared to the two urban groups. In both LREB and Islamic groups, body mass and femur length was positively correlated with δ13C values, and δ15N shows a positive correlation with left humerus shape in the LREB Urban sample. Conclusions The low stature and cross-sectional properties of the Rural Islamic group are most likely an indicator of greater physiological stress, potentially due to poorer diet. Positive correlations between measures of body size and δ13C values further suggest that greater access to C4 resources improved diet quality. Alternatively, this relationship could indicate greater body size among migrants from areas where individuals consumed more C4 resources

Clinical evaluation of antiseptic mouth rinses to reduce salivary load of SARS-CoV-2

Most public health measures to contain the COVID-19 pandemic are based on preventing the pathogen spread, and the use of oral antiseptics has been proposed as a strategy to reduce transmission risk. The aim of this manuscript is to test the efficacy of mouthwashes to reduce salivary viral load in vivo. This is a multi-centre, blinded, parallel-group, placebo-controlled randomised clinical trial that tests the effect of four mouthwashes (cetylpyridinium chloride, chlorhexidine, povidone-iodine and hydrogen peroxide) in SARS-CoV-2 salivary load measured by qPCR at baseline and 30, 60 and 120 min after the mouthrinse. A fifth group of patients used distilled water mouthrinse as a control. Eighty-four participants were recruited and divided into 12-15 per group. There were no statistically significant changes in salivary viral load after the use of the different mouthwashes. Although oral antiseptics have shown virucidal effects in vitro, our data show that salivary viral load in COVID-19 patients was not affected by the tested treatments. This could reflect that those mouthwashes are not effective in vivo, or that viral particles are not infective but viral RNA is still detected by PCR. Viral infectivity studies after the use of mouthwashes are therefore required

SARS-CoV-2 viral load in nasopharyngeal swabs is not an independent predictor of unfavorable outcome

The aim was to assess the ability of nasopharyngeal SARS-CoV-2 viral load at first patient’s hospital evaluation to predict unfavorable outcomes. We conducted a prospective cohort study including 321 adult patients with confirmed COVID-19 through RT-PCR in nasopharyngeal swabs. Quantitative Synthetic SARS-CoV-2 RNA cycle threshold values were used to calculate the viral load in log10 copies/mL. Disease severity at the end of follow up was categorized into mild, moderate, and severe. Primary endpoint was a composite of intensive care unit (ICU) admission and/or death (n = 85, 26.4%). Univariable and multivariable logistic regression analyses were performed. Nasopharyngeal SARS-CoV-2 viral load over the second quartile (≥ 7.35 log10 copies/mL, p = 0.003) and second tertile (≥ 8.27 log10 copies/mL, p = 0.01) were associated to unfavorable outcome in the unadjusted logistic regression analysis. However, in the final multivariable analysis, viral load was not independently associated with an unfavorable outcome. Five predictors were independently associated with increased odds of ICU admission and/or death: age ≥ 70 years, SpO2, neutrophils > 7.5 × 103/µL, lactate dehydrogenase ≥ 300 U/L, and C-reactive protein ≥ 100 mg/L. In summary, nasopharyngeal SARS-CoV-2 viral load on admission is generally high in patients with COVID-19, regardless of illness severity, but it cannot be used as an independent predictor of unfavorable clinical outcome