Topical Brazilian propolis improves corneal wound healing and inflammation in rats following alkali burns

Abstract Background The objective of this study was to investigate the effects of the Brazilian Scaptotrigona sp propolis, a widely used folk medicine, in corneal wound healing and inflammation. Methods Corneal epithelial defects of 1 mm in diameter were made in the right eyes of Wistar male adult rats by cauterization with silver nitrate sticks. Subsequently, they were divided in two groups (n = 40 rats/group): Brazilian propolis (BP) group was topically treated with a microemulsion containing 1% Brazilian propolis; vehicle (VH) group received the same formulation without propolis. The epithelial defect area was photographed and measured at t = 0 (wound induction), and after 12, 24, 48 and 120 h of treatment. The inflammatory response was evaluated based on counting of neutrophils. Epithelial regeneration rates were determined based on Ki-67 expression in basal epithelial cells. Comparisons were made using the Kruskal-Wallis and the Mann–Whitney U test. Results The BP group presented both smaller epithelial defect areas at 12, 24 and 48 h and fewer corneal infiltrating neutrophils at 24 and 48 h (P < 0.01) than the VH group. These effects were associated with more pervasive Ki-67 staining in the BP group at 12 and 24 h (P < 0.05). Conclusions Topically applied BP accelerated wound healing and reduced the inflammatory response to silver nitrate-induced corneal alkali burns in rats.

Expression analysis and in silico characterization of intronic long noncoding RNAs in renal cell carcinoma: emerging functional associations

Abstract Background Intronic and intergenic long noncoding RNAs (lncRNAs) are emerging gene expression regulators. The molecular pathogenesis of renal cell carcinoma (RCC) is still poorly understood, and in particular, limited studies are available for intronic lncRNAs expressed in RCC Methods Microarray experiments were performed with custom-designed arrays enriched with probes for lncRNAs mapping to intronic genomic regions. Samples from 18 primary RCC tumors and 11 nontumor adjacent matched tissues were analyzed. Meta-analyses were performed with microarray expression data from three additional human tissues (normal liver, prostate tumor and kidney nontumor samples), and with large-scale public data for epigenetic regulatory marks and for evolutionarily conserved sequences. Results A signature of 29 intronic lncRNAs differentially expressed between RCC and nontumor samples was obtained (false discovery rate (FDR) <5%). A signature of 26 intronic lncRNAs significantly correlated with the RCC five-year patient survival outcome was identified (FDR <5%, p-value ≤0.01). We identified 4303 intronic antisense lncRNAs expressed in RCC, of which 22% were significantly (p <0.05) cis correlated with the expression of the mRNA in the same locus across RCC and three other human tissues. Gene Ontology (GO) analysis of those loci pointed to 'regulation of biological processes’ as the main enriched category. A module map analysis of the protein-coding genes significantly (p <0.05) trans correlated with the 20% most abundant lncRNAs, identified 51 enriched GO terms (p <0.05). We determined that 60% of the expressed lncRNAs are evolutionarily conserved. At the genomic loci containing the intronic RCC-expressed lncRNAs, a strong association (p <0.001) was found between their transcription start sites and genomic marks such as CpG islands, RNA Pol II binding and histones methylation and acetylation. Conclusion Intronic antisense lncRNAs are widely expressed in RCC tumors. Some of them are significantly altered in RCC in comparison with nontumor samples. The majority of these lncRNAs is evolutionarily conserved and possibly modulated by epigenetic modifications. Our data suggest that these RCC lncRNAs may contribute to the complex network of regulatory RNAs playing a role in renal cell malignant transformation.

Microbial Translocation Is Associated with Extensive Immune Activation in Dengue Virus Infected Patients with Severe Disease

Background: Severe dengue virus (DENV) disease is associated with extensive immune activation, characterized by a cytokine storm. Previously, elevated lipopolysaccharide (LPS) levels in dengue were found to correlate with clinical disease severity. In the present cross-sectional study we identified markers of microbial translocation and immune activation, which are associated with severe manifestations of DENV infection. Methods: Serum samples from DENV-infected patients were collected during the outbreak in 2010 in the State of Sa˜o Paulo, Brazil. Levels of LPS, lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14) and IgM and IgG endotoxin core antibodies were determined by ELISA. Thirty cytokines were quantified using a multiplex luminex system. Patients were classified according to the 2009 WHO classification and the occurrence of plasma leakage/shock and hemorrhage. Moreover, a (non-supervised) cluster analysis based on the expression of the quantified cytokines was applied to identify groups of patients with similar cytokine profiles. Markers of microbial translocation were linked to groups with similar clinical disease severity and clusters with similar cytokine profiles. Results: Cluster analysis indicated that LPS levels were significantly increased in patients with a profound pro-inflammatory cytokine profile. LBP and sCD14 showed significantly increased levels in patients with severe disease in the clinical classification and in patients with severe inflammation in the cluster analysis. With both the clinical classification and the cluster analysis, levels of IL-6, IL-8, sIL-2R, MCP-1, RANTES, HGF, G-CSF and EGF were associated with severe disease. Conclusions: The present study provides evidence that both microbial translocation and extensive immune activation occur during severe DENV infection and may play an important role in the pathogenesis.

Validação do escore de meningite bacteriana em crianças com pleocitose em São Paulo, Brasil

Introdução. A maioria das crianças com pleocitose tem meningite viral, não bacteriana. O Escore de Meningite Bacteriana (EMB) é uma ferramenta clínica para identificar crianças com pleocitose de muito baixo risco para meningite bacteriana (MB). Crianças são consideradas de muito baixo risco para MB se, de acordo com o EMB, não tiverem nenhum dos cinco parâmetros - presença de convulsões à admissão, neutrófilos no liquor > 1000/mm3, neutrófilos no sangue >10000/mm3, coloração de Gram demonstrando bactérias e proteínas no LCR > 80 mg/dL. Objetivo. Avaliar a performance do EMB para distinguir meningite viral de bacteriana em crianças com pleocitose. Métodos. Foi realizado um estudo de coorte retrospectivo em um departamento de emergência de hospital secundário de ensino. Foram identificadas crianças entre 1 mês e 15 anos incompletos entre 2001 e 2011 com pleocitose (mais de 10 células/mm3 no liquor). Foram calculados sensibilidade, especificidade e valores preditivos positivo e negativo do EMB. Resultados. Foram identificados 497 crianças com meningite, sendo 43 (8,6%; intervalo de confiança [IC] 95% 7-13%) com MB e 454 (91,4%; IC 95% 87-93%) com meningite asséptica. Das 208 crianças de muito baixo risco para MB de acordo com o EMB, nenhuma tinha MB (sensibilidade 100%, IC 95% 92-100%; especificidade 62%, IC 95% 57- 67%; valor preditivo negativo 100%; IC 95% 98-100%). Conclusões. O EMB identificou com acurácia elevada crianças de muito baixo risco para MB. Este modelo pode ser utilizado para ajuda na diferenciação entre MB e meningite viral, principalmente em regiões com cobertura incompleta vacinal para meningococo e pneumococo.

The intronic long noncoding RNA ANRASSF1 recruits PRC2 to the RASSF1A promoter, reducing the expression of RASSF1A and increasing cell proliferation

The down-regulation of the tumor-suppressor gene RASSF1A has been shown to increase cell proliferation in several tumors. RASSF1A expression is regulated through epigenetic events involving the polycomb repressive complex 2 (PRC2); however, the molecular mechanisms modulating the recruitment of this epigenetic modifier to the RASSF1 locus remain largely unknown. Here, we identify and characterize ANRASSF1, an endogenous unspliced long noncoding RNA (lncRNA) that is transcribed from the opposite strand on the RASSF1 gene locus in several cell lines and tissues and binds PRC2. ANRASSF1 is transcribed through RNA polymerase II and is 5'-capped and polyadenylated; it exhibits nuclear localization and has a shorter half-life compared with other lncRNAs that bind PRC2. ANRASSF1 endogenous expression is higher in breast and prostate tumor cell lines compared with non-tumor, and an opposite pattern is observed for RASSF1A. ANRASSF1 ectopic overexpression reduces RASSF1A abundance and increases the proliferation of HeLa cells, whereas ANRASSF1 silencing causes the opposite effects. These changes in ANRASSF1 levels do not affect the RASSF1C isoform abundance. ANRASSF1 overexpression causes a marked increase in both PRC2 occupancy and histone H3K27me3 repressive marks, specifically at the RASSF1A promoter region. No effect of ANRASSF1 overexpression was detected on PRC2 occupancy and histone H3K27me3 at the promoter regions of RASSF1C and the four other neighboring genes, including two well-characterized tumor suppressor genes. Additionally, we demonstrated that ANRASSF1 forms an RNA/DNA hybrid and recruits PRC2 to the RASSF1A promoter. Together, these results demonstrate a novel mechanism of epigenetic repression of the RASSF1A tumor suppressor gene involving antisense unspliced lncRNA, in which ANRASSF1 selectively represses the expression of the RASSF1 isoform overlapping the antisense transcript in a location-specific manner. In a broader perspective, our findings suggest that other non-characterized unspliced intronic lncRNAs transcribed in the human genome might contribute to a location-specific epigenetic modulation of genes.

Eduardo Zamacois: attività editoriale e opera letteraria

Eduardo Zamacois (Pinar del Río, 1873- Buenos Aires, 1971) was a main actor of the spanish’s literature and edition movement from the first third of the 20th century. He was the founder of magazines that had a big impact like “Germinal”, introductive of the sicalipsis (“La Vida Galante”) or so innovative that deserved an special chapter in the history of literature (“El Cuento Semanal” e “Los Contemporáneos”). With this work, it is intended to recreate the most significant stages from his autobiography adventure including the new information that offers his non before published letters exchange with his last sentimental partner. As a writer, his work was very popular in his homeland, translated to the world and reissued in Ibero-America. His literature work is based in three different phases. He began with the use of gallant literature (with books like La enferma, Punto negro) and took an adventure with the pays of mystery and irony (El otro, El misterio de un hombre pequñito, La opinion ajena) to finally focused in a narrative style of a realistic kind, that includes social critique (Las raíces). His last published novel in Spain is a portrait of Madrid during its long siege by the nationalist troops (El asedio de Madrid), a profound tribute to the people’s heroism and a declaration of love to the capital, that was his place of residence for a long time. While his exile, that took him to Cuba, New York and Buenos Aires, he worked in the radio, the dubbing industry and in finishing his most detailed bibliography, Un hombre que se va, a valuable document of that time.

Las estructuras antropológicas de lo imaginario en "Las Rosas de Hércules", de Tomás Morales

Programa de doctorado: Literatura y Teoría de la Literatura

Proposta di ritraduzione dei primi capitoli di A Song of Ice and Fire e analisi della resa italiana di Sergio Altieri

Lo scopo di questa tesi è quello di presentare una proposta di ritraduzione per i primi tre capitoli del primo romanzo di A Song of Ice and Fire. La scelta di questo tema è stata dettata dalla contestata traduzione ufficiale di quest'opera alla quale sono personalmente appassionato, e dal conseguente desiderio di sfruttare le conoscenze acquisite durante i miei studi per ovviare ai problemi della resa di Sergio Altieri. Il lavoro è sviluppato in tre capitoli e un'appendice. Nel primo capitolo vengono analizzate le problematiche generali legate alla traduzione. Viene inoltre presentata una sinossi all'opera per semplificare la lettura della tesi. Nel secondo capitolo vengono esaminati i problemi principali della traduzione di Altieri. Il terzo capitolo è dedicato al confronto tra l'opera originale, la traduzione ufficiale e la proposta di ritraduzione dei primi capitoli. In appendice è presente la mia traduzione integrale dei tre capitoli del romanzo.

Guidelines for the use and interpretation of assays for monitoring autophagy

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.