Central carbon metabolism exhibits unique characteristics during the handling of fungal patterns by monocyte-derived dendritic cells.

Monocyte-derived dendritic cells (MDDCs) are key players in the defense against fungal infection because of their outstanding capacity for non-opsonic phagocytosis and phenotypic plasticity. Accordingly, MDDCs rewire metabolism to meet the energetic demands for microbial killing and biomass synthesis required to restore homeostasis. It has been commonplace considering the metabolic reprogramming a mimicry of the Warburg effect observed in tumor cells. However, this may be an oversimplification since the offshoots of glycolysis and the tricarboxylic acid (TCA) cycle are connected in central carbon metabolism. Zymosan, the external wall of Saccharomyces cerevisiae, contains ß-glucan and α-mannan chains that engage the C-type lectin receptors dectin-1/2 and Toll-like receptors. This makes it an optimal fungal surrogate for experimental research. Using real-time bioenergetic assays and [U-13C]glucose labeling, central hubs connected to cytokine expression were identified. The pentose phosphate pathway (PPP) exhibited a more relevant capacity to yield ribose-5-phosphate than reducing equivalents of NADPH, as judged from the high levels of isotopologues showing 13C-labeling in the ribose moiety and the limited contribution of the oxidative arm of the PPP to the production of ROS by NADPH oxidases (NOX). The finding of 13C-label in the purine ring and in glutathione unveiled the contribution of serine-derived glycine to purine ring and glutathione synthesis. Serine synthesis also supported the TCA cycle. Zymosan exhausted NAD+ and ATP, consistent with intracellular consumption and/or extracellular export. Poly-ADP-ribosylated proteins detected in the nuclear fractions of MDDCs did not show major changes upon zymosan stimulation, which suggests its dependence on constitutive Fe(II)/2-oxoglutarate-dependent demethylation of 5-methylcytosine by TET translocases and/or demethylation of histone H3 lysine 27 by JMJD demethylases rather than on NOX activities. These results disclose a unique pattern of central carbon metabolism following fungal challenge, characterized by the leverage of glycolysis offshoots and an extensive recycling of NAD+ and poly(ADP-ribose).

Safety and efficacy of ribociclib plus letrozole in patients with HR+, HER2- advanced breast cancer: Results from the Spanish sub-population of the phase 3b CompLEEment-1 trial.

BACKGROUND: Breast cancer is the most common malignancy and the second leading cause of cancer-related mortality in Spanish women. Ribociclib in combination with endocrine therapy (ET) has shown superiority in prolonging survival in patients with hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) advanced breast cancer (ABC) vs. ET alone. METHODS: CompLEEment-1 is a single-arm, open-label phase 3b trial evaluating ribociclib plus letrozole in a broad population of patients with HR+, HER2- ABC. The primary endpoints were safety and tolerability. Here we report data for Spanish patients enrolled in CompLEEment-1. RESULTS: A total of 526 patients were evaluated (median follow-up: 26.97 months). Baseline characteristics showed a diverse population with a median age of 54 years. At study entry, 56.5% of patients had visceral metastases and 8.7% had received prior chemotherapy for advanced disease. Rates of all-grade and Grade ≥3 adverse events (AEs) were 99.0% and 76.2%, respectively; 21.3% of patients experienced a serious AE, and 15.8% of AEs led to treatment discontinuation. AEs of special interest of neutropenia, increased alanine aminotransferase, increased aspartate aminotransferase and QTcF prolongation occurred in 77.8%, 14.8%, 11.4% and 4.0% of patients, respectively. Patients aged >70 years experienced increased rates of all-grade and Grade ≥3 neutropenia and anemia. Efficacy results were consistent with the global study. CONCLUSIONS: Results from Spanish patients enrolled in CompLEEment-1 are consistent with global data showing efficacy and a manageable safety profile for ribociclib plus letrozole treatment in patients with HR+, HER2- ABC, including populations of interest (NCT02941926). TRIAL REGISTRATION: ClinicalTrials.gov NCT02941926.

Incidence of and factors associated with SARS-CoV-2 infection among people living with HIV in Southern Spain after one year of pandemic.

Whether people living with HIV (PLWH) are at greater risk of acquiring SARS-CoV-2 infection is currently unknown. Prospective serologic studies may allow seroincidence analyses, where all infections are accurately identified. Because of this, we evaluated the incidence of associated factors with and the clinical outcome of SARS-CoV-2 infection in PLWH in Southern Spain. This prospective cohort study included PLWH from a Tertiary University Hospital in Southern Spain. Patients were enrolled in the study if (1) they had attended as outpatients our Unit from 1 August 2019 to 8 February 2020 and (2) had two subsequent evaluations from 9 February 2020 to 4 March 2021. SARS-CoV-2 infections were diagnosed by PCR, antigen detection or serology. Seven hundred and nine PLWH were included in the study. Of them, 55 [7.8%, 95% confidence interval (95% CI) 5.9%-9.9%] patients developed SARS-CoV-2 infection. Between 18 May and 29 November 2020, the rate of seroconversion was 5.3% (95% CI: 3.1%-9.0%) for the general population in the area of Seville and 2.3% (95% CI: 1.3%-2.6%) for PLWH in this study (p = .001). After multivariable analysis, adjusted by age, sex, and risk factors for HIV infection, active tobacco use and CDC stage, active tobacco smoking was the only factor independently associated with lower risk of SARS-Cov-2 infection [Incidence rate ratio: 0.29 (95% CI 0.16-0.55) p < .001]. In conclusion, the incidence of SARS-CoV-2 infection among PLWH in Southern Spain during the ongoing pandemic was lower than that reported for the general population in the same area.

Enfoque intersectorial y responsabilidad social universitaria en la gestión del conocimiento sobre actividad física y salud / Intersector approach and university social responsibly in managing knowledge about physical activity and health

Introducción: La inactividad física constituye uno de los principales factores de riesgo de morbimortalidad a nivel mundial y son causa fundamental de las enfermedades no transmisibles. En cambio, la actividad física regular durante el curso de la vida proporciona beneficios para la salud física y mental. No obstante, aun resultan insuficientes las acciones intersectoriales que potencien alianzas estratégicas, en aras de evidenciar el rol de la actividad física como componente del estilo de vida saludable. Objetivo: Identificar la responsabilidad social del Centro de Estudio para la Actividad Física, el Deporte y la Promoción de la Salud, en la formación de capacidades para la producción, la transferencia, la diseminación y la aplicación de conocimientos en el contexto de la actividad física para la salud. Desarrollo: La inactividad física, por su estrecha relación comórbida con la obesidad y el sobrepeso, la diabetes, la hipertensión, la hipercolesterolemia, entre otras, da al traste con los principales problemas de salud durante el curso de la vida, lo cual reclama de un enfoque sistémico, integrador e intersectorial, así como de la formación de recursos humanos competentes para ello. Conclusiones: El Centro de Estudios para la Actividad Física, el Deporte y la Promoción de Salud evidencia su pertinencia académica e investigativa, al devenir en un actor clave en la producción, la distribución y el uso del conocimiento sobre la actividad física para la salud, y articular las necesidades del contexto social y el proyecto universitario, alineados con las prioridades nacionalmente establecidas(AU)

Introduction: Physical inactivity is one of the main risk factors for morbidity and mortality worldwide and its fundamental cause is non-communicable diseases. In contrast, regular physical activity during the span of life provides physical and mental health benefits. However, there are still insufficient intersector actions for strengthening strategic alliances, in view of demonstrating the role of physical activity as a component of the healthy lifestyle. Objective: To identify the social responsibility of the University of Physical Culture and Sports Sciences, particularly its Center for Studies on Physical Activity, Sports and Health Promotion, in the formation of capacities for knowledge production, transfer, dissemination and application in the context of physical activity for health. Development: Physical inactivity, due to its close comorbid relationship with obesity and overweight, diabetes, hypertension, hypercholesterolemia, among others, disrupts into the main health problems during the lifespan, which demands a systemic, integrative and intersector approach, as well as the training of competent human resources for it. Conclusions: The Center for Studies on Physical Activity, Sports and Health Promotion proves its academic and research relevance, insofar as it becomes a key stakeholder in the production, distribution and use of knowledge about physical activity for health, and articulates the needs of the social context and the university project, aligned with nationally established priorities(AU)

Diagnóstico de la práctica de ejercicio físico en el municipio Guanabacoa / Diagnosis of the practice of physical exercise in the Guanabacoa municipality

Resumen En el mundo actual, las personas realizan ejercicios físicos como vía para mejorar su estado de salud, condición física, imagen, utilización constructiva del ocio y calidad de vida. Diferentes son los estudios que demuestran la efectividad del ejercicio físico en el organismo, también hacen referencia a la importancia de la dosificación de los mismos para evitar lesiones y enfermedades en los practicantes. Esto se convierte en uno de los pasos vitales cuando se practica, pues a partir de ahí se marca la meta que se persigue y según la dosificación, que se aplique, serán los resultados. El objetivo de esta investigación fue diagnosticar el estado actual de la práctica de ejercicio físico en el municipio Guanabacoa. El tipo de estudio empleado fue no experimental-descriptivo. Los métodos utilizados fueron entre los teóricos, el analítico-sintético; dentro de los empíricos la encuesta y el análisis documental y como estadístico-matemático, la estadística descriptiva. Los resultados revelan la necesidad de que especialistas capacitados dirijan la práctica de ejercicios físicos en el municipio Guanabacoa. Se comprueba el desconocimiento por parte de los especialistas, en la dosificación y planificación de los ejercicios físicos.

Abstract In today's world, people perform physical exercises as a way to improve their health status, physical condition, image, constructive use of leisure and quality of life. Different studies that demonstrate the effectiveness of physical exercise in the body, also refer to the importance of dosing them to avoid injuries and diseases in practitioners. This becomes one of the vital steps when we are going to practice them, because from there we mark the goal that we pursue and according to the dosage that we apply, the results will be. The objective of this research was to diagnose the current state of the practice of physical exercise in the Guanabacoa municipality. The type of study used was not experimental-descriptive. The methods used were among the theorists, the analytical-synthetic; within the empirical surveys and documentary analysis and as statistical-mathematical, descriptive statistics. The results reveal the need for trained specialists to direct the practice of physical exercises in the municipality of Guanabacoa. The lack of knowledge on the part of the specialists in the dosage and planning of the physical exercises is checked.

Polypropylene Carbonate-Based Adaptive Buffer Layer for Stable Interfaces of Solid Polymer Lithium Metal Batteries.

Solid polymer electrolytes (SPEs) have the potential to enhance the safety and energy density of lithium batteries. However, poor interfacial contact between the lithium metal anode and SPE leads to high interfacial resistance and low specific capacity of the battery. In this work, we present a novel strategy to improve this solid-solid interface problem and maintain good interfacial contact during battery cycling by introducing an adaptive buffer layer (ABL) between the Li metal anode and SPE. The ABL consists of low molecular-weight polypropylene carbonate , poly(ethylene oxide) (PEO), and lithium salt. Rheological experiments indicate that ABL is viscoelastic and that it flows with a higher viscosity compared to PEO-only SPE. ABL also has higher ionic conductivity than PEO-only SPE. In the presence of ABL, the interface resistance of the Li/ABL/SPE/LiFePO4 battery only increased 20% after 150 cycles, whereas that of the battery without ABL increased by 117%. In addition, because ABL makes a good solid-solid interface contact between the Li metal anode and SPE, the battery with ABL delivered an initial discharge specific capacity of >110 mA·h/g, which is nearly twice that of the battery without ABL, which is 60 mA·h/g. Moreover, ABL is able to maintain electrode-electrolyte interfacial contact during battery cycling, which stabilizes the battery Coulombic efficiency.

Polymorphisms in Receptors Involved in Opsonic and Nonopsonic Phagocytosis, and Correlation with Risk of Infection in Oncohematology Patients.

High-risk hematological malignancies are a privileged setting for infection by opportunistic microbes, with invasive mycosis being one of the most serious complications. Recently, genetic background has emerged as an unanticipated risk factor. For this reason, polymorphisms for genes encoding archetypal receptors involved in the opsonic and nonopsonic clearance of microbes, pentraxin-3 (PTX3) and Dectin-1, respectively, were studied and correlated with the risk of infection. Fungal, bacterial, and viral infections were registered for a group of 198 patients with high-risk hematological malignancies. Polymorphisms for the pentraxin-3 gene (PTX3) showed a significant association with the risk of fungal infection by Candida spp. and, especially, by Aspergillus spp. This link remained even for patients undergoing antifungal prophylaxis, thus demonstrating the clinical relevance of PTX3 in the defense against fungi. CLEC7A polymorphisms did not show any definite correlation with the risk of invasive mycosis, nor did they influence the expression of Dectin-1 isoforms generated by alternative splicing. The PTX3 mRNA expression level was significantly lower in samples from healthy volunteers who showed these polymorphisms, although no differences were observed in the extents of induction elicited by bacterial lipopolysaccharide and heat-killed Candidaalbicans, thus suggesting that the expression of PTX3 at the start of infection may influence the clinical outcome. PTX3 mRNA expression can be a good biomarker to establish proper antifungal prophylaxis in immunodepressed patients.

Seis décadas de avances y retos para la promoción de la salud en Cuba / Six decades of advances and challenges for Cuban health's promotion

La prioridad otorgada por el gobierno cubano a la promoción de la salud, es una realidad que se hace evidente en los resultados sociales alcanzados en las últimas seis décadas, con una tendencia ascendente y demuestra la coherencia política y sinergias entre estrategias de inversión social y enfoques de salud pública. Presentar los avances y desafíos de la promoción de la salud, en el contexto social cubano, en las últimas décadas, es objetivo del artículo. El método, deviene del análisis crítico de la información disponible en fuentes bibliográficas, documentos y entrevistas. Los resultados se expresaron en indicadores de salud y desarrollo, publicados, disponibles y referenciados, así como en ejemplos de proyectos y estrategias comunitarias. Los resultados develaron que las políticas gubernamentales, sectoriales y locales, han permitido que el sistema de salud disponga de herramientas valiosas para desempeñar su encargo en la promoción de la salud, con acciones tangibles; se identificaron logros como mejora de indicadores y retos como la atención a la política de viviendas. Las diferentes aristas de la participación social mostraron su influencia en los resultados alcanzados, aunque imbuirlas de nuevos aires es una urgencia. Las consideraciones finales declaran que el enfoque de la promoción de la salud en Cuba, avanza, en la medida que lo permiten las complejidades y amenazas a las que se enfrenta el sistema político y social, en un país resiliente y de valores humanos demostrados a escala nacional e internacional(AU)

The priority given by the revolutionary government to health promotion for all Cubans from 1959 to 2016 is a reality which is evidenced in the social results achieved in the last six decades with an upward tendency, and as a proof of political coherence and synergy among the social investment strategies and the public health approach. The objective of this article is to present the advances and challenges of health promotion in the Cuban social context in the last sixty years. The used method becomes from the critical analysis of available information of indexed bibliographic sources, documents and interviews to key informants. The results were expressed in the health and development indicators that were published and available in the references, as the examples of community projects and strategies. The results showed that governmental, sectorial and local policies allowed the health system to have at its disposal valuable tools to perform with concrete actions its assignment related to health promotion. Some achievements as the improvement of indicators were identified, but also challenges as the attention to the housing policy. The different areas of social participation showed their influence in the achieved results, although it urges to fuel them with new approaches. As final considerations, it is declared that the approach for health promotion in Cuba is moving forward, as much as the complexities and threats faced by the political and social system let it in the context of a resilient country with high human values that have been proved nationally and internationally(AU)

Fungal pattern receptors down-regulate the inflammatory response by a cross-inhibitory mechanism independent of interleukin-10 production.

Cyclic AMP regulatory element binding protein and signal transducer and activator of transcription 3 (STAT3) may control inflammation by several mechanisms, one of the best characterized is the induction of the expression of the anti-inflammatory cytokine interleukin-10 (IL-10). STAT3 also down-regulates the production of pro-inflammatory cytokines induced by immunoreceptor tyrosine-based activation motif (ITAM)-coupled receptors, a mechanism termed cross-inhibition. Because signalling via ITAM-dependent mechanisms is a hallmark of fungal pattern receptors, STAT3 activation might be involved in the cross-inhibition associated with invasive fungal infections. The fungal surrogate zymosan produced the phosphorylation of Y705-STAT3 and the expression of Ifnb1 and Socs3, but did not induce the interferon (IFN)-signature cytokines Cxcl9 and Cxcl10 in bone marrow-derived dendritic cells. Unlike lipopolysaccharide (LPS), zymosan induced IL-10 and phosphorylated Y705-STAT3 to a similar extent in Irf3 and Ifnar1 knockout and wild-type mice. Human dendritic cells showed similar results, although the induction of IFNB1 was less prominent. These results indicate that LPS and zymosan activate STAT3 through different routes. Whereas type I IFN is the main effector of LPS effect, the mechanism involved in Y705-STAT3 phosphorylation by zymosan is more complex, cannot be associated with type I IFN, IL-6 or granulocyte-macrophage colony-stimulating factor, and seems dependent on several factors given that it was partially inhibited by the platelet-activating factor antagonist WEB2086 and high concentrations of COX inhibitors, p38 mitogen-activate protein kinase inhibitors, and blockade of tumour necrosis factor-α function. Altogether, these results indicate that fungal pattern receptors share with other ITAM-coupled receptors the capacity to produce cross-inhibition through a mechanism involving STAT3 and induction of SOCS3 and IL-10, but that cannot be explained through type I IFN signalling.

Association of Performance Status and Pain in Metastatic Bone Pain Management in the Spanish Clinical Setting.

INTRODUCTION: Bone metastasis is the most common cause of cancer-related pain, and metastatic bone pain (MBP) is not only severe but also progressive in many patients. The aim of this study was to investigate the association between pain management and performance status in patients with metastatic bone cancer in the Spanish clinical setting. METHODS: A 3-month follow-up prospective, epidemiologic, multicenter study was conducted in 579 patients to assess the evolution of their performance, the impact of pain control on sleep and functionality, and the degree of pain control according to analgesic treatment. RESULTS: In patients with MBP, Eastern Cooperative Oncology Group (ECOG) status (1.5 ± 0.7-1.3 ± 0.7 and 1.3 ± 0.8; p < 0.001) and pain (6.5 ± 1.4-2.8 ± 1.9 and 2.1 ± 1.9; p < 0.001) improved significantly from baseline to months 1 and 3, as did functionality and sleep, after a treatment change consisting of increasing the administration of opioids. Evolution of ECOG and pain were closely related. ECOG and pain outcomes were significantly more favorable in patients treated with opioids versus non-opioid treatment, and in patients who did not need rescue medication versus those who did. CONCLUSIONS: MBP is currently poorly managed in Spain. ECOG improvement is closely and directly related to pain management in MBP. Opioid treatment and a lack of requirements for rescue medication are associated with better ECOG and pain outcomes in MBP patients. FUNDING: Mundipharma Pharmaceuticals S.L.

Pharmacological inhibition of eicosanoids and platelet-activating factor signaling impairs zymosan-induced release of IL-23 by dendritic cells.

The engagement of the receptors for fungal patterns induces the expression of cytokines, the release of arachidonic acid, and the production of PGE2 in human dendritic cells (DC), but few data are available about other lipid mediators that may modulate DC function. The combined antagonism of leukotriene (LT) B4, cysteinyl-LT, and platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) inhibited IL23A mRNA expression in response to the fungal surrogate zymosan and to a lower extent TNFA (tumor necrosis factor-α) and CSF2 (granulocyte macrophage colony-stimulating factor) mRNA. The combination of lipid mediators and the lipid extract of zymosan-conditioned medium increased the induction of IL23A by LPS (bacterial lipopolysaccharide), thus suggesting that unlike LPS, zymosan elicits the production of mediators at a concentration enough for optimal response. Zymosan induced the release of LTB4, LTE4, 12-hydroxyeicosatetraenoic acid (12-HETE), and PAF C16:0. DC showed a high expression and detectable Ser663 phosphorylation of 5-lipoxygenase in response to zymosan, and a high expression and activity of LPCAT1/2 (lysophosphatidylcholine acyltransferase 1 and 2), the enzymes that incorporate acetate from acetyl-CoA into choline-containing lysophospholipids to produce PAF. Pharmacological modulation of the arachidonic acid cascade and the PAF receptor inhibited the binding of P-71Thr-ATF2 (activating transcription factor 2) to the IL23A promoter, thus mirroring their effects on the expression of IL23A mRNA and IL-23 protein. These results indicate that LTB4, cysteinyl-LT, and PAF, acting through their cognate G protein-coupled receptors, contribute to the phosphorylation of ATF2 and play a central role in IL23A promoter trans-activation and the cytokine signature induced by fungal patterns.

The unfolded protein response and the phosphorylations of activating transcription factor 2 in the trans-activation of il23a promoter produced by ß-glucans.

Current views on the control of IL-23 production focus on the regulation of il23a, the gene encoding IL-23 p19, by NF-κB in combination with other transcription factors. C/EBP homologous protein (CHOP), X2-Box-binding protein 1 (XBP1), activator protein 1 (AP1), SMAD, CCAAT/enhancer-binding protein (C/EBPß), and cAMP-response element-binding protein (CREB) have been involved in response to LPS, but no data are available regarding the mechanism triggered by the fungal mimic and ß-glucan-containing stimulus zymosan, which produces IL-23 and to a low extent the related cytokine IL-12 p70. Zymosan induced the mobilization of CHOP from the nuclear fractions to phagocytic vesicles. Hypha-forming Candida also induced the nuclear disappearance of CHOP. Assay of transcription factor binding to the il23a promoter showed an increase of Thr(P)-71-Thr(P)-69-activating transcription factor 2 (ATF2) binding in response to zymosan. PKC and PKA/mitogen- and stress-activated kinase inhibitors down-regulated Thr(P)-71-ATF2 binding to the il23a promoter and il23a mRNA expression. Consistent with the current concept of complementary phosphorylations on N-terminal Thr-71 and Thr-69 of ATF2 by ERK and p38 MAPK, MEK, and p38 MAPK inhibitors blunted Thr(P)-69-ATF2 binding. Knockdown of atf2 mRNA with siRNA correlated with inhibition of il23a mRNA, but it did not affect the expression of il12/23b and il10 mRNA. These data indicate the following: (i) zymosan decreases nuclear proapoptotic CHOP, most likely by promoting its accumulation in phagocytic vesicles; (ii) zymosan-induced il23a mRNA expression is best explained through coordinated κB- and ATF2-dependent transcription; and (iii) il23a expression relies on complementary phosphorylation of ATF2 on Thr-69 and Thr-71 dependent on PKC and MAPK activities.

Polarization of the innate immune response by prostaglandin E2: a puzzle of receptors and signals.

Eicosanoids tailor the innate immune response by supporting local inflammation and exhibiting immunomodulatory properties. Prostaglandin (PG) E2 is the most abundant eicosanoid in the inflammatory milieu due to the robust production elicited by pathogen-associated molecular patterns on cells of the innate immune system. The different functions and cell distribution of E prostanoid receptors explain the difficulty encountered thus far to delineate the actual role of PGE2 in the immune response. The biosynthesis of eicosanoids includes as the first step the Ca(2+)- and kinase-dependent activation of the cytosolic phospholipase A2, which releases arachidonic acid from membrane phospholipids, and later events depending on the transcriptional regulation of the enzymes of the cyclooxygenase routes, where PGE2 is the most relevant product. Acting in an autocrine/paracrine manner in macrophages, PGE2 induces a regulatory phenotype including the expression of interleukin (IL)-10, sphingosine kinase 1, and the tumor necrosis factor family molecule LIGHT. PGE2 also stabilizes the suppressive function of myeloid-derived suppressor cells, inhibits the release of IL-12 p70 by macrophages and dendritic cells, and may enhance the production of IL-23. PGE2 is a central component of the inflammasome-dependent induction of the eicosanoid storm that leads to massive loss of intravascular fluid, increases the mortality rate associated with coinfection by Candida ssp. and bacteria, and inhibits fungal phagocytosis. These effects have important consequences for the outcome of infections and the polarization of the immune response into the T helper cell types 2 and 17 and can be a clue to develop pharmacological tools to address infectious, autoimmune, and autoinflammatory diseases.

The response of human macrophages to ß-glucans depends on the inflammatory milieu.

BACKGROUND: ß-glucans are fungal cell wall components that bind to the C-type lectin-like receptor dectin-1. Polymorphisms of dectin-1 gene are associated with susceptibility to invasive fungal infection and medically refractory ulcerative colitis. The purpose of this study has been addressing the response of human macrophages to ß-glucans under different conditions mimicking the composition of the inflammatory milieu in view of the wide plasticity and large range of phenotypical changes showed by these cells, and the relevant role of dectin-1 in several pathophysiological conditions. PRINCIPAL FINDINGS: Serum-differentiated macrophages stimulated with ß-glucans showed a low production of TNFα and IL-1ß, a high production of IL-6 and IL-23, and a delayed induction of cyclooxygenase-2 and PGE2 biosynthesis that resembled the responses elicited by crystals and those produced when phagosomal degradation of the phagocytic cargo increases ligand access to intracellular pattern recognition receptors. Priming with a low concentration of LPS produced a rapid induction of cyclooxygenase-2 and a synergistic release of PGE2. When the differentiation of the macrophages was carried out in the presence of M-CSF, an increased expression of dectin-1 B isoform was observed. In addition, this treatment made the cells capable to release arachidonic acid in response to ß-glucan. CONCLUSIONS: These results indicate that the macrophage response to fungal ß-glucans is strongly influenced by cytokines and microbial-derived factors that are usual components of the inflammatory milieu. These responses can be sorted into three main patterns i) an elementary response dependent on phagosomal processing of pathogen-associated molecular patterns and/or receptor-independent, direct membrane binding linked to the immunoreceptor tyrosine-based activation motif-bearing transmembrane adaptor DNAX-activating protein 12, ii) a response primed by TLR4-dependent signals, and iii) a response dependent on M-CSF and dectin-1 B isoform expression that mainly signals through the dectin-1 B/spleen tyrosine kinase/cytosolic phospholipase A2 route.

Sirtuin 1 is a key regulator of the interleukin-12 p70/interleukin-23 balance in human dendritic cells.

Stimulation of human dendritic cells with the fungal surrogate zymosan produces IL-23 and a low amount of IL-12 p70. Trans-repression of il12a transcription, which encodes IL-12 p35 chain, by proteins of the Notch family and lysine deacetylation reactions have been reported as the underlying mechanisms, but a number of questions remain to be addressed. Zymosan produced the location of sirtuin 1 (SIRT1) to the nucleus, enhanced its association with the il12a promoter, increased the nuclear concentration of the SIRT1 co-substrate NAD(+), and decreased chromatin accessibility in the nucleosome-1 of il12a, which contains a κB-site. The involvement of deacetylation reactions in the inhibition of il12a transcription was supported by the absence of Ac-Lys-14-histone H3 in dendritic cells treated with zymosan upon coimmunoprecipitation of transducin-like enhancer of split. In contrast, we did not obtain evidence of a possible effect of SIRT1 through the deacetylation of c-Rel, the central element of the NF-κB family involved in il12a regulation. These data indicate that an enhancement of SIRT1 activity in response to phagocytic stimuli may reduce the accessibility of c-Rel to the il12a promoter and its transcriptional activation, thus regulating the IL-12 p70/IL-23 balance and modulating the ongoing immune response.

Reversible Inter- and Intra-Microgel Cross-Linking using Disulfides.

Thermoresponsive hydrogel nanoparticles composed of poly(N-isopropylmethacrylamide) (pNIPMAm) and the disulfide-based cross-linker N,N'-bis(acryloyl)cystamine (BAC) have been prepared using a redox-initiated, aqueous precipitation polymerization approach, leading to improved stability of the disulfide bond compared to traditional thermally-initiated methods. The resultant particles demonstrate complete erosion in response to reducing conditions or thiol competition. This stands in contrast to the behavior of thermally-initiated particles, which retain a cross-linked network following disulfide cleavage due to uncontrolled chain-branching and self-cross-linking side reactions. The synthetic strategy has also been combined with the non-degradable cross-linker N,N-methylenebisacrylamide (BIS) to generate "co-cross-linked" pNIPMAm-BAC-BIS microgels. These particles are redox-responsive, swell upon BAC cross-link scission and present reactive thiols. This pendant thiol functionality was demonstrated to be useful for conjugation of thiol-reactive probes and in reversible network formation by assembling particles cross-linked by disulfide linkages.

Notch- and transducin-like enhancer of split (TLE)-dependent histone deacetylation explain interleukin 12 (IL-12) p70 inhibition by zymosan.

The fungal analog zymosan induces IL-23 and low amounts of IL-12 p70. This study addresses the molecular mechanisms underlying this cytokine pattern in human monocyte-derived dendritic cells. The transcriptional regulation of il23a, one of the chains of IL-23, depended on the activation of c-Rel and histone H3 phosphorylation, as judged from the association of c-Rel with the il23a promoter and the correlation between IL-23 production and Ser-10-histone H3 phosphorylation. Consistent with its reduced ability to produce IL-12 p70, zymosan induced a transient occupancy of the il12a promoter by c-Rel, blocked the production of IL-12 p70 and the transcription of il12a induced by other stimuli, and triggered the expression and nuclear translocation of the transcriptional repressors of the Notch family hairy and enhancer of split (Hes)-1, Hes5, hairy/enhancer-of-split related with YRPW motif protein (Hey)-1, and transducin-like enhancer of split (TLE). Zymosan also induced the interaction of Hes1 and TLE with histone H3 phosphorylated on Ser-10 and deacetylated on Lys-14. Inhibition of class III histone deacetylases increased the production of IL-12 p70 and partially blunted the inhibitory effect of zymosan on the production of IL-12 p70 elicited by LPS and IFN-γ. These results indicate that the selective induction of IL-23 by ß-glucans is explained by the activation of c-Rel associated with Ser-10-histone H3 phosphorylation in the il23a promoter mediated by mitogen- and stress-activated kinase and/or protein kinase A and inhibition of il12a transcription by a mechanism involving activation of several corepressors with the ability to bind TLE and to promote histone deacetylation.

Eicosanoids in the innate immune response: TLR and non-TLR routes.

The variable array of pattern receptor expression in different cells of the innate immune system explains the induction of distinct patterns of arachidonic acid (AA) metabolism. Peptidoglycan and mannan were strong stimuli in neutrophils, whereas the fungal extract zymosan was the most potent stimulus in monocyte-derived dendritic cells since it induced the production of PGE(2), PGD(2), and several cytokines including a robust IL-10 response. Zymosan activated kappaB-binding activity, but inhibition of NF-kappaB was associated with enhanced IL-10 production. In contrast, treatments acting on CREB (CRE binding protein), including PGE(2), showed a direct correlation between CREB activation and IL-10 production. Therefore, in dendritic cells zymosan induces il10 transcription by a CRE-dependent mechanism that involves autocrine secretion of PGE(2), thus unraveling a functional cooperation between eicosanoid production and cytokine production.

The induction of IL-10 by zymosan in dendritic cells depends on CREB activation by the coactivators CREB-binding protein and TORC2 and autocrine PGE2.

Stimulation of human monocyte-derived dendritic cells with the yeast extract zymosan is characterized by a predominant production of IL-10 and a strong induction of cyclooxygenase-2, but the molecular mechanisms underlying this response are only partially understood. To address this issue, the activation of transcription factors that may bind to the il10 proximal promoter was studied. Binding activity to Sp1, Sp3, NF-Y, and cAMP response element (CRE) sites was detected in the nuclear extracts of dendritic cells; however these binding activities were not influenced by zymosan. No binding activity to Stat1, Stat3, and c/EBP sites was detected. Notably, zymosan activated kappaB-binding activity, but inhibition of NF-kappaB was associated with enhanced IL-10 production. In sharp contrast, treatments acting on CREB (CRE binding protein), including 8-Br-cAMP, PGE(2), and inhibitors of PKA, COX, and glycogen-synthase kinase-3beta showed a direct correlation between CREB activation and IL-10 production. Zymosan induced binding of both P-CREB and CREB-binding protein (CBP) to the il10 promoter as judged from chromatin immunoprecipitation assays, whereas negative results were obtained with Ab reactive to Sp1, Sp3, c-Maf, and NF-Y. Zymosan also induced nuclear translocation of the CREB coactivator transducer of regulated CREB activity 2 (TORC2) and interaction of TORC2 with P-CREB coincidental with the association of CREB to the il10 promoter. Altogether, our data show that zymosan induces il10 transcription by a CRE-dependent mechanism that involves autocrine secretion of PGE(2) and a network of interactions of PKA, MAP/ERK, glycogen-synthase kinase-3beta, and calcineurin, which regulate CREB transcriptional activity by binding the coactivators CBP and TORC2 and inhibiting CBP interaction with other transcription factors.

Efecto del extracto acuoso de Aloe vera (L) N L Burm sobre indicadores lipídicos de suero de conejo / Effect of the water extract from Aloe vera (L) N L Burm on rabbit serum lipids

Introducción: trabajos realizados con extracto acuoso y gel hojas de Aloe vera (L) N L Burm, administrados a ratones en un modelo de hiperlipidemia aguda inducida con tritón, demostraron un efecto protector al incremento de los lípidos. Objetivos: estudiar el efecto de la administración a dosis repetida de extracto acuoso de A. vera sobre los indicadores lipídicos de suero de conejo. Métodos: conejos machos Nueva Zelandia fueron tratados con extractos acuosos de A. vera a las dosis de 17 y 34 mg/kg de peso corporal, sobre la base de sólidos totales durante 6 semanas, también se incluyó un grupo control sin tratamiento, el peso corporal, colesterol total, HDL-colesterol, LDL colesterol y triglicéridos fueron determinados al inicio y al final del período del experimento. Resultados: se encontró una disminución significativa del colesterol, LDL colesterol y triglicéridos en suero de los animales tratados con la dosis de 34 mg/kg de extracto acuoso al final del experimento, pero el HDL-colesterol no mostró variaciones. Conclusiones: la administración de extracto acuoso de A. vera a conejo durante 6 semanas causó una disminución significativa de los lípidos en suero.

Introduction: the research works conducted with water extract and gel from Aloe vera (L.) N. L. Burm., administered to mice in a triton-induced acute hyperlipidemia pattern, showed a protective effect against the increase of lipids. Objectives: to study the effect of administration of water extract from A. vera at repeated doses on the lipid indicators of rabbit serum. Methods: male New Zealand rabbits were treated with water extracts from Aloe vera at doses of 17 and 34 mg/kg of bodyweight, on the basis of whole solids for 6 weeks. An untreated control group was included. Body weight, total cholesterol, HDL-cholesterol, LDL cholesterol and triglycerides were determined at the beginning and at the end of the experimental period. Results: it was found that cholesterol, LDL cholesterol and triglycerides significantly decreased in serum of animals treated with 34 mg/kg dose of the water extract at the end of the experiment, however, the HDL-cholesterol did not show any variation. Conclusions: the administration of water extract from Aloe vera to a rabbit for 6 weeks significantly reduced serum lipids.