Mediterranean diet impact on cardiovascular diseases: a narrative review

: Cardiovascular disease (CVD) accounts for more than 17 million deaths per year worldwide. It has been estimated that the influence of lifestyle on CVD mortality amounts to 13.7% for smoking, 13.2% for poor diet, and 12% for inactive lifestyle. These results deeply impact both the healthy status of individuals and their skills in working. The impact of CVD on productivity loss accounts for the 24% in total costs for CVD management.Mediterranean diet (MedD) can positively impact on natural history of CVD. It is characterized by a relatively high consumption of inexpensive and genuine food such as cereals, vegetables, legumes, nuts, fish, fresh fruits, and olive oil as the principal source of fat, low meat consumption and low-to-moderate consumption of milk, dairy products, and wine.Its effects on cardiovascular health are related to the significant improvements in arterial stiffness. Peripheral artery disease, coronary artery disease, and chronic heart failure are all positively influenced by the MedD. Furthermore, MedD lowers the risk of sudden cardiac death due to arrhythmias.The present narrative review aims to analyze the effects of MedD on CVD

The Drosophila Helicase MLE Targets Hairpin Structures in Genomic Transcripts

<div><p>RNA hairpins are a common type of secondary structures that play a role in every aspect of RNA biochemistry including RNA editing, mRNA stability, localization and translation of transcripts, and in the activation of the RNA interference (RNAi) and microRNA (miRNA) pathways. Participation in these functions often requires restructuring the RNA molecules by the association of single-strand (ss) RNA-binding proteins or by the action of helicases. The Drosophila MLE helicase has long been identified as a member of the MSL complex responsible for dosage compensation. The complex includes one of two long non-coding RNAs and MLE was shown to remodel the roX RNA hairpin structures in order to initiate assembly of the complex. Here we report that this function of MLE may apply to the hairpins present in the primary RNA transcripts that generate the small molecules responsible for RNA interference. Using stocks from the Transgenic RNAi Project and the Vienna Drosophila Research Center, we show that MLE specifically targets hairpin RNAs at their site of transcription. The association of MLE at these sites is independent of sequence and chromosome location. We use two functional assays to test the biological relevance of this association and determine that MLE participates in the RNAi pathway.</p></div

RNase treatment strongly reduces MLE signal at the integration site of the plasmid.

<p><b>(A)</b> Left panel, MLE staining of polytene chromosomes from male larvae expressing a dsRNA targeting <i>Hrb87F</i> after induction with <i>Actin5C-GAL4</i>. The incubation of the salivary glands in RNase A (RNase A) perturbs MLE localization at the integration site of the plasmid while in the absence of RNase A (ctrl) the MLE signal is still highly enriched. The white arrows indicate the plasmid integration site. In the right panel is a detail of the region marked by the arrows. <b>(B)</b> Quantitative analysis of fluorescence levels. MLE signal at the integration site of the plasmid, expressed in terms of corrected total band fluorescence (CTBF), is significantly reduced after RNase A treatment (p value <0.001). The analysis was performed on 33 polytene chromosomes treated with RNase A and 11 control chromosomes.</p

Rm62 RNA helicase is not enriched at sites of hairpin RNA transcription.

<p>Left panel, Rm62 staining of polytene chromosomes from female larvae expressing a dsRNA targeting <i>Hrb87F</i> after induction with <i>Actin5C-GAL4</i>. The white arrows indicate the plasmid integration site. The right panel shows a detail of the region marked by the arrows.</p

MLE targets shRNA at their site of transcription.

<p>MLE staining of polytene chromosomes from female larvae expressing short hairpin RNAs after induction with <i>Actin5C-GAL4</i>.</p

MLE mutation affects RNAi efficiency.

<p><b>(A)</b> Wing phenotypes from female flies in which Notch dsRNA expression is induced by <i>C96-GAL4</i>. Class1: wild type; class 2: missing margin bristles and absent notching; class 3: moderate notching; class 4: extensive notching; class 5: missing most of the wing margins; class 6: complete lack of margins and reduced wing blade. The chart on the right side of the figure represents the <i>Notch</i> RNAi wing phenotypes distribution in <i>wild type</i>, <i>mle</i> homozygous mutant and <i>mle</i> heterozygous mutant background. The results are the sum of three to six independent crosses per genotype. <b>(B)</b> Wing phenotypes from female flies in which Egfr dsRNA expression is induced by <i>ms1096-GAL4</i>. Class1: wild type; class 2: all the veins present; class 3: absent anterior cross vein (acv) or presence of a partial longitudinal vein; class 4: absent acv plus one partial longitudinal vein; class 5: absent acv plus two partial longitudinal veins; class 6: acv vein absent plus one longitudinal vein absent plus one partial longitudinal vein; class 7: most of the veins absent. The chart on the right side of the figure represents the <i>Egfr</i> RNAi wing phenotypes distribution respectively in <i>wild type</i> and <i>mle</i> homozygous mutant background. The results are the sum of four to five independent crosses per genotype.</p

MLE localization at the integration site of the plasmid does not require the MSL complex.

<p><b>(A)</b> Left panel, Polytene chromosomes from male larvae expressing a dsRNA targeting <i>Hrb87F</i> (<i>Hrb87F</i> RNAi) under the induction of <i>Actin5C-GAL4</i>. MSL1, MSL3 and MOF paint the X chromosome but are absent at the integration site of the plasmid indicated by the white arrows. In the right panel is a detail of the region marked by the arrows. <b>(B)</b> Polytene chromosomes from female larvae expressing a dsRNA targeting <i>Hrb87F</i> (<i>Hrb87F</i> RNAi) following induction with <i>Actin5C-GAL4</i> and larvae in which the production of the dsRNA is not induced (ctrl).</p

MLE is enriched at the plasmid integration site when transcription of the transgene is active.

<p>Left panel, Polytene chromosomes from male larvae expressing a dsRNA targeting <i>Hrb87F</i> (<i>Hrb87F</i> RNAi) under the induction of <i>Actin5C-GAL4</i> or larvae in which the production of the dsRNA is not activated (ctrl). MLE paints the X chromosome in both samples and is enriched at the integration site of the plasmid only in <i>Hrb87F</i> RNAi larvae. The white arrows indicate the plasmid integration site. In the right panel is a detail of the region marked by the arrows.</p

MLE targets sites of dsRNA transcription in a sequence and chromosome location independent manner.

<p><b>(A)</b> MLE staining of polytene chromosomes from female larvae expressing either a <i>mof</i> dsRNA or an <i>Hrb87F</i> dsRNA construct from a pValium1 insertion of TRiP line collection following induction with <i>Actin5C-GAL4</i>. <b>(B)</b> GAL4 and MLE staining of polytene chromosomes from female larvae expressing dsRNA constructs integrated respectively at Chr2L 30B3 (<i>Jil1</i> RNAi) and Chr2L 22A5 (<i>Hp1</i> RNAi) and induced by <i>Actin5C-GAL4</i>.</p