MicroRNAs (miRNAs), small noncoding RNAs modulating messenger RNA (mRNA) and protein expression, have emerged as key regulatory molecules in chronic liver diseases, whose end stage is hepatic ...fibrosis, a major global health burden. Pharmacological strategies for prevention or treatment of hepatic fibrosis are still limited, what makes it necessary to establish a better understanding of the molecular mechanisms underlying its pathogenesis. In this context, we have recently shown that cyclooxygenase-2 (COX-2) expression in hepatocytes restricts activation of hepatic stellate cells (HSCs), a pivotal event in the initiation and progression of hepatic fibrosis. Here, we evaluated the role of COX-2 in the regulation of a specific set of miRNAs on a mouse model of CCl4 and bile duct ligation (BDL)-induced liver fibrosis. Our results provide evidence that COX-2 represses miR-23a-5p and miR-28-5p expression in HSC. The decrease of miR-23a-5p and miR-28-5p expression promotes protection against fibrosis by decreasing the levels of pro-fibrogenic markers α-SMA and COL1A1 and increasing apoptosis of HSC. Moreover, we demonstrate that serum levels of miR-28-5p are decreased in patients with chronic liver disease. These results suggest a protective effect exerted by COX-2-derived prostanoids in the process of hepatofibrogenesis.
•hCOX-2 Tg mice have attenuated fibrosis by modulating miR-23a and miR-28 in HSCs.•Hepatic PGE2 production downregulates miR-23a and miR-28 in HSCs.•Downregulation of miR-23a and miR-28a decreases activation of HSC.•Decreased proliferation and increased apoptosis of HSC lead to fibrosis resolution.
The pathogenic mechanisms underlying the progression of non-alcoholic fatty liver disease (NAFLD) are not fully understood. In this study, we aimed to assess the relationship between endoplasmic ...reticulum (ER) stress and autophagy in human and mouse hepatocytes during NAFLD. ER stress and autophagy markers were analyzed in livers from patients with biopsy-proven non-alcoholic steatosis (NAS) or non-alcoholic steatohepatitis (NASH) compared with livers from subjects with histologically normal liver, in livers from mice fed with chow diet (CHD) compared with mice fed with high fat diet (HFD) or methionine-choline-deficient (MCD) diet and in primary and Huh7 human hepatocytes loaded with palmitic acid (PA). In NASH patients, significant increases in hepatic messenger RNA levels of markers of ER stress (activating transcription factor 4 (ATF4), glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP)) and autophagy (BCN1) were found compared with NAS patients. Likewise, protein levels of GRP78, CHOP and p62/SQSTM1 (p62) autophagic substrate were significantly elevated in NASH compared with NAS patients. In livers from mice fed with HFD or MCD, ER stress-mediated signaling was parallel to the blockade of the autophagic flux assessed by increases in p62, microtubule-associated protein 2 light chain 3 (LC3-II)/LC3-I ratio and accumulation of autophagosomes compared with CHD fed mice. In Huh7 hepatic cells, treatment with PA for 8 h triggered activation of both unfolding protein response and the autophagic flux. Conversely, prolonged treatment with PA (24 h) induced ER stress and cell death together with a blockade of the autophagic flux. Under these conditions, cotreatment with rapamycin or CHOP silencing ameliorated these effects and decreased apoptosis. Our results demonstrated that the autophagic flux is impaired in the liver from both NAFLD patients and murine models of NAFLD, as well as in lipid-overloaded human hepatocytes, and it could be due to elevated ER stress leading to apoptosis. Consequently, therapies aimed to restore the autophagic flux might attenuate or prevent the progression of NAFLD.
The molecular hallmark of the Ewing's family of tumors is the presence of balanced chromosomal translocations, leading to the formation of chimerical transcription factors (that is, EWS/FLI1) that ...play a pivotal role in the pathogenesis of Ewing's tumors by deregulating gene expression. We have recently demonstrated that DAX1 (NR0B1), an orphan nuclear receptor that was not previously implicated in cancer, is induced by the EWS/FLI1 oncoprotein and is highly expressed in Ewing's tumors, suggesting that DAX1 is a biologically relevant target of EWS/FLI1-mediated oncogenesis. In this study we demonstrate that DAX1 is a direct transcriptional target of the EWS/FLI1 oncoprotein through its binding to a GGAA-rich region in the DAX1 promoter and show that DAX1 is a key player of EWS/FLI1-mediated oncogenesis. DAX1 silencing using an inducible model of RNA interference induces growth arrest in the A673 Ewing's cell line and severely impairs its capability to grow in semisolid medium and form tumors in immunodeficient mice. Gene expression profile analysis demonstrated that about 10% of the genes regulated by EWS/FLI1 in Ewing's cells are DAX1 targets, confirming the importance of DAX1 in Ewing's oncogenesis. Functional genomic analysis, validated by quantitative RT-PCR, showed that genes implicated in cell-cycle progression, such as CDK2, CDC6, MCM10 or SKP2 were similarly regulated by EWS/FLI1 and DAX1. These findings indicate that DAX1 is important in the pathogenesis of the Ewing's family of tumors, identify new functions for DAX1 as a cell-cycle progression regulator and open the possibility to new therapeutic approaches based on DAX1 function interference.
Mangrove swamps and forests cover over 137,000 km2 distributed latitudinally among subtropical zones, 7% of which are in Brazil, with a greater density in the country's northernmost region. ...Considering that the community of Myxomycetes recorded for this environment is hardly known, three areas located in the state of Maranhão were investigated. Two field trips were conducted, one at the beginning of the rainy season and another during the dry season. In each area, two plots (125 m2) equidistant 100 m apart from each other were surveyed. In these areas, standing dead tree trunks and dead branches still attached to the mother plant that were above the tideline, were examined. On these same occasions, samples of the aerial litter and from the cortex of living trees (Rhizophora) were collected for the preparation of moist chambers cultures. Twenty-one specimens were obtained from field and moist chambers, belonging to 11 species, distributed in nine genera and five families. Seven species are new records from Maranhão. There was a predominance of r-strategist (73%) over K-strategist (27%) species. Cribraria violacea, Comatricha tenerrima, Echinostelium minutum, and Fuligo septica are new worldwide records from mangrove environments, and Oligonema flavidum is reported for the first time from Brazil.
In this study, we identified candidate genes associated with the reproductive success of
Varroa destructor
in
Apis mellifera
brood cells at a key point in time, the 4
th
day after host cell capping. ...First-life cycle reproducing (R) and non-reproducing (NR) mites obtained by applying a semi-field rearing protocol were analyzed by RNA-Seq and qPCR. We observed a general under-expression of genes associated with metabolism and a few over-expressed genes putatively involved in signal transduction and DNA-binding transcription factor activity in R compared to NR mites. Specifically, PTCH1 and AP-1 genes, associated with embryonic segmentation and apoptosis, emerged as the best candidates for reproductive success in R mites and the early abortion of reproduction in NR mites. We also detected differentially transcribed long non-coding RNAs between R and NR mites with
cis
target genes annotated as regulators of replication, transcription, and translation pathways. Some of these genes were specifically associated with embryonic development and apoptosis. We discuss putative metabolic pathways associated with
V. destructor
reproduction to provide novel information for developing innovative and environmentally friendly mite control strategies.
Cyclooxygenases (COX-1 and 2) catalyze the first step in prostanoid biosynthesis. They are implicated in homeostatic processes with an important role in inflammation and carcinogenesis. In the liver, ...COX-2 expression is restricted to proliferation or dedifferentiation situations. The COX-2 promoter contains numerous CpG islands that, when hypermethylated, result in transcriptionally silencing thus regulating the growth of carcinoma cells. In this work, we investigated whether a correlation exists between COX-2 expression and methylation signatures at the 5'region of the gene in hepatoma cell lines and human hepatocellular carcinoma (HCC). We also examined the acetylation status of the COX-2 promoter and the effects of histone deacetylase (HDAC) inhibitors on COX-2 expression. Our results suggest a significant association between reduced COX-2 expression and promoter hypermethylation of COX-2 and histone deacetylation in some hepatoma cell lines and in HCC. Treatment with demethylating agents or HDAC inhibitors restored the expression of COX-2. Moreover, in an HCC cohort, a statistically significant inverse association was observed between COX-2 mRNA levels and promoter methylation. In agreement with these data, a reduction of overall survival of the patients was observed after decreased COX-2 expression by promoter hypermethylation and histone H3 hypoacetylation.
Identification of the "bean smut" reported in 1998 in abstracts from two conferences was later disseminated by a Plant Disease Note; citations in books, papers, and blogs; and in several official ...sites, including databases curated by the United States Department of Agriculture and Embrapa-Brazil. After seeing the illustration of the syndrome in 2002, the need became clear for a review of the so-called bean smut. Field collections indicated that it is common in no-till bean and soybean farming in Brazil. Our studies revealed that the "bean smut" attributed to Ustilago sp. or "Ustilago phaseoli" and, later, to "Microbotryum phaseoli" is not a real smut but is Physarum cinereum (Physaraceae, Physarales, Myxomycetes), sporulating superficially on leaves, stems, and pods of dry bean and soybean. To unravel this imbroglio, we produced detailed morphological documentation supported by molecular treatment. This will correct the spread and further incorporation of an error in the literature based upon mistaken taxonomical work related to a plant-associated nonpathogenic organism.
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