AimsMicroRNAs (miRNAs) play an important role in the pathogenesis of structural alterations of the failing heart through their ability to regulate negatively the expression levels of genes that govern the process of adaptive and maladaptive cardiac remodelling. We studied whether LV reverse remodelling after CRT was associated with changes of circulating miRNAs in patients with heart failure (HF) and dyssynchrony. Methods and resultsA prospective, non-randomized self-control trial was performed in 81 patients with HF eligible for CRT. At baseline, to select the HF miRNA profile, we evaluated the expression of 84 miRNAs (implicated in the pathogenesis of structural alterations of the failing heart) in three groups of patients: healthy subjects (healthy group, n ¼ 15); patients with HF (HF group, n ¼ 81); and patients without HF matched for age, sex, and concomitant disease with HF patients (control group, n ¼ 60). At 12 months, the selected miRNA profile was evaluated in plasma from responder (n ¼ 55) and non-responder HF patients (n ¼ 26) to CRT. In the test cohort, the HF patients were characterized by lower expression of 48 miRNAs (all P , 0.04) as compared with healthy subjects. In the validation cohort, the HF patients were characterized by lower expression of 24 miRNAs (all P , 0.03) as compared with control patients. At 12 months, 55 patients (68%) were considered responders and 26 non-responders to CRT (32%). Responders showed an increase in expression of 19 miRNAs (all P , 0.03) compared with baseline expression, whereas in the non-responders we observed an increase of six miRNAs (all P , 0.05) compared with baseline expression. At follow-up, miRNAs were differentially expressed between responders and non-responders. The responders were characterized by higher expression of five miRNAs (miRNA-26b-5p, miRNA-145-5p, miRNA-92a-3p, miRNA-30e-5p, and miRNA-29a-3p; P , 0.01 for all) as compared with non-responders. ConclusionsIn responders, reverse remodelling is associated with favourable changes in miRNAs that regulate cardiac fibrosis, apoptosis, and hypertrophy.--

MicroRNAs are small non-coding RNAs that modulate gene expression at post-transcriptional level, playing a crucial role in cell differentiation and development. Recently, some reports have shown that a limited number of mammalian microRNAs are also involved in anti-viral defense. In this study, the analysis of the hepatitis B virus (HBV) genome by the computer program MiRanda led to the identification of seven sites that are potential targets for human liver microRNAs. These sites were found to be clustered in a 995-bp segment within the viral polymerase ORF and the overlapping surface antigen ORF, and conserved among the most common HBV subtypes. The HBV genomic targets were then subjected to a validation test based on cultured hepatic cells (HepG2, HuH-7 and PLC/PRF/5) and luciferase reporter genes. In this test, one of the selected microRNAs, hsa-miR-125a-5p, was found to interact with the viral sequence and to suppress the reporter activity markedly. The microRNA was then shown to interfere with the viral translation, down-regulating the expression of the surface antigen. Overall, these results support the emerging concept that some mammalian microRNAs play a role in virus-host interaction. Furthermore, they provide the basis for the development of new strategies for anti-HBV intervention.

Hepatocellular carcinoma (HCC) is the third cause of cancer-related deaths worldwide. Sorafenib is the only approved drug for patients with advanced HCC but has shown limited activity. microRNAs (miRs) have been involved in several neoplasms including HCC suggesting their use or targeting as good tools for HCC treatment. The purpose of this study was to identify novel approaches to sensitize HCC cells to sorafenib through miRs. miR-423-5p was validated as positive regulator of autophagy in HCC cell lines by transient transfection of miR and anti-miR molecules. miR-423-5p expression level was evaluated by real-time polymerase chain reaction (PCR) in sera collected from 39 HCC patients before and after treatment with sorafenib. HCC cells were cotreated with sorafenib and miR-423-5p and the effects on cell cycle, apoptosis, and autophagy were evaluated. Secretory miR-423-5p was upregulated both in vitro and in vivo by sorafenib treatment and its increase was correlated with response to therapy since 75% of patients in which an increase of secretory miR423-5p was found were in partial remission or stable disease after 6 moths from the beginning of therapy. HCC cells transfected with miR-423-5p showed an increase of cell percentage in S-phase of cell cycle paralleled by a similar increase of autophagic cells evaluated at both fluorescence activated cell sorter (FACS) and transmission electron microscopy. Our results suggest the miR423-5p can be used as a useful tool to predict response to sorafenib in HCC patients and is involved in autophagy regulation in HCC cells.

Ras proteins are highly related GTPases that have key roles in regulating growth, differentiation and tumorigenesis. Genetargeting experiments have shown that, out of the three mammalian ras genes, only K-ras is essential for normal mouse embryogenesis, and that mice deprived of H-ras and/or N-ras show no major phenotype. We generated mice (HrasKI) in which the K-ras gene had been modified to encode H-Ras protein. HrasKI mice produce undetectable amounts of K-Ras but-in contrast to mice homozygous for a null K-ras allele-they are born at the expected mendelian frequency, indicating that H-Ras can be substituted for K-Ras in embryonic development. However, adult HrasKI mice show dilated cardiomyopathy associated with arterial hypertension. Our results show that K-Ras can be replaced by H-Ras in its essential function in embryogenesis, and indicate that K-Ras has a unique role in cardiovascular homeostasis.

To study in HBsAg chronic carriers the expression of liver hsa-miR-125a-5p and its correlation with liver HBV-DNA values and clinical presentation, 27 consecutive Caucasian, HBsAg/anti-HBe/HBV-DNA-positive patients who were naive to nucleos(t)ide analogues and interferon therapy and had no marker of HCV, HDV or HIV infection and no history of alcohol intake were enrolled. For each patient, liver HBV DNA and liver hsa-miR-125a-5p were quantified by real-time PCR in relation to β-globin DNA or RNU6B, respectively. Liver fibrosis and necroinflammation were graded by applying Ishak's scoring system. Liver hsa-miR-125a-5p was detected in all patients enrolled and a correlation between its concentration and liver HBV DNA was demonstrated (p<0.0001). Higher liver hsa-miR-125a-5p concentrations were observed in patients with HBV-DNA plasma level >103 IU/ml (p<0.02), in those with HAI >6 (p = 0.02) and those with fibrosis score >2 (p<0.02) than in patients with lower scores. Higher HBV-DNA liver concentrations were found in patients with abnormal AST (p = 0.005) and ALT serum levels (p = 0.05), in those with serum HBV DNA higher than 10E3 IU/mL (p = 0.001) and those with fibrosis score >2 (p = 0.02) than in patients with a lower load. By multivariate logistic regression analysis, liver hsa-miR-125a-5p was identified as an independent predictor of disease progression: O.R. = 4.21, C.I. 95%  = 1.08–16.43, p<0.05, for HAI >6; O.R. = 3.12, C.I. 95%  = 1.17–8.27, p<0.05, for fibrosis score >2. In conclusion, in HBsAg/anti-HBe-positive patients, the liver hsa-miR-125a-5p level correlated with liver and plasma HBV-DNA values and was associated to a more severe disease progression.

Oxidative stress has been proposed to be an important factor in the pathogenesis of Alzheimer's disease (AD), playing a central role in amyloid β-protein (Aβ) generation and neuronal apoptosis. Oxidative damage directly correlates with the presence of Aβ deposits. Aβ and oxidative stress jointly induce neuronal death, Aβ deposits, gliosis, and memory impairment in AD. In order to counteract AD neurodegeneration, the inhibition of the vicious cycle of Aβ generation and oxidation is an attractive therapeutic strategy, and antiamyloidogenic and antioxidant herbal drugs could represent an alternative and valid approach. In this context, an alcoholic extract from Laurus nobilis leaves (LnM) and seven fractions obtained therefrom were of interest. All extracts prepared through extractive and chromatographic techniques were phytochemically studied by chromatographic techniques including gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS(n)). The potential antioxidant efficacy of the obtained fractions was screened by DPPH(•) and ABTS(•+) assays, as well as specific assay media characterized from the presence of highly reactive ROS and RNS species (ROO(•), OH(•), O2(•-), and NO). In order to evaluate the preparation of safe and nontoxic extracts, MTT, SRB, and LDH assays toward SH-5YSY and SK-N-BE(2)-C human neuronal cell lines, as well as on C6 mouse glial cell line, were performed. The apoptosis-inducing properties by spectroscopic evaluation of the extracts' ability to activate caspase-3 and by a DNA fragmentation assay were also investigated. Data thus obtained allowed us to state the absence of toxic effects induced by phenolic-rich fractions (LnM, LnM-1, LnM-1a, LnM-1b, and LnM-2c), which at the same time exerted significant cytoprotective and antioxidant responses in hydrogen peroxide and Aβ(25-35)-fragment-oxidized cell systems. The potential antiamyloidogenic efficacy of Laurus nobilis leaf polar extracts in the Aβ(25-35) fragment oxidized cell systems was further analyzed by Congo red staining.

Edited by Tamas DalmayKeywords: MicroRNA Virus RNA interference Hepatitis HIV-1 Papillomavirus a b s t r a c t MicroRNAs are small non-coding RNAs that modulate gene expression at post-transcriptional level, playing a crucial role in cell differentiation and development. Recently, some reports have shown that a limited number of mammalian microRNAs also display antiviral effects. This article summarizes the data in the field paying a special attention to the conservation of the microRNA target sequences in the viral populations. This issue is relevant both for the evaluation of the biological significance of the antiviral effects and for the development of microRNA-based strategies for antiviral intervention.

MicroRNAs are small non-coding RNAs that regulate gene expression at post-transcriptional level by inhibiting translation of complementary mRNAs and/or targeting them for degradation. MicroRNAs play crucial roles in development, cell differentiation, and apoptosis. In addition, recent studies indicate that they are important regulators of virus-host interactions. MicroRNA-125a is a homolog of C. elegans lin-4, the first discovered microRNA, shown to dictate the onset of larval stages in the nematode. In this review, we focus on the gene structure of microRNA-125a, its evolution, its expression pattern in mammalian organs and tissues, and its functional targets. Overall, the available data indicate that microRNA-125a plays crucial roles both in development and in the adult tissues. In fact, it (1) contributes to the control of phase transitions in development and/or cell differentiation; (2) regulates the expression of several target proteins that are involved in cell proliferation, apoptosis, and migration; (3) interferes with the expression of the hepatitis B virus surface antigen in liver cells, thus counteracting viral replication. These findings suggest that delivery of microRNA-125a mimics or treatments that modulate its cellular expression may be valuable tools for the development of new therapeutic strategies for human diseases, including cancer and viral hepatitis B.