Criteri di buona governance in Leader: l\u2019autovalutazione dei Gruppi di Azione Locale

l principale obiettivo di questo lavoro \ue8 proporre una metodologia per l\u2019autovalutazione della capacit\ue0 del Gruppi di Azione Locale (Gal) di elaborare e gestire una strategia locale Leader secondo principi di buona governance. L\u2019approccio proposto, inizialmente sviluppato e testato in via preliminare con tecniche di ricerca qualitativa \ue8 qui perfezionato e integrato con il Common Assessment Framework (Caf), un modello di autovalutazione in uso da tempo presso organizzazioni pubbliche in Europa. Il sistema proposto si basa su un set d\u2019indicatori specifici di prestazione (performance) e di percezione del Gal capaci di correlare i principali elementi di una buona governance alle specificit\ue0 dell\u2019approccio Leader. Questo strumento pu\uf2 contribuire a migliorare l\u2019attivit\ue0 di monitoraggio gestionale e indirizzare criticamente le attivit\ue0 del Gal

Cryptides Identified in Human Apolipoprotein B as New Weapons to Fight Antibiotic Resistance in Cystic Fibrosis Disease

Chronic respiratory infections are the main cause of morbidity and mortality in cystic fibrosis (CF) patients, and are characterized by the development of multidrug resistance (MDR) phenotype and biofilm formation, generally recalcitrant to treatment with conventional antibiotics. Hence, novel eective strategies are urgently needed. Antimicrobial peptides represent new promising therapeutic agents. Here, we analyze for the first time the ecacy of three versions of a cryptide identified in human apolipoprotein B (ApoB, residues 887-922) towards bacterial strains clinically isolated from CF patients. Antimicrobial and anti-biofilm properties of ApoB-derived cryptides have been analyzed by broth microdilution assays, crystal violet assays, confocal laser scanning microscopy and scanning electron microscopy. Cell proliferation assays have been performed to test cryptide eects on human host cells. ApoB-derived cryptides have been found to be endowed with significant antimicrobial and anti-biofilm properties towards Pseudomonas and Burkholderia strains clinically isolated from CF patients. Peptides have been also found to be able to act in combination with the antibiotic ciprofloxacin, and they are harmless when tested on human bronchial epithelial mesothelial cells. These findings open interesting perspectives to cryptide applicability in the treatment of chronic lung infections associated with CF disease

A Comparison among Analytical Methods to Assess Fatty Acids and Conjugated Linoleic Acids (CLA) Content and Repeatability of Ruminant Faeces

Methods to determine fatty acids (FAs) and CLA contents of faeces should limit isomerisation, provide a good repeatability of the measures, avoid the use of harmful substances. Three methods of FAs extraction from faeces for GC analysis were compared: Est-DFtol, based on extraction and esterification of FAs contained in dry faeces using Na-methoxide, methanolic-HCl and toluene as solvent; Est-EEtol, based on acid-base extraction and esterification of FAs on the faecal ether extract (EE), using toluene as solvent; and AEst-EEhept, based on an acid catalyzed esterification of FAs contained in EE, using n-heptane as solvent. Faeces were collected from bulls receiving 0, 8 and 80 g/d of rumen protected CLA (rpCLA). The faeces of 9 bulls (3 for each dose) were analysed in triplicates by each method. Methods were compared by linear regression. The measurements performed with Est-EEtol and AEst-EEhept regressed against those of Est-DFtol, evidenced, in particularly for CLA isomers and their sum, positive intercepts and slopes significantly lower than the unity. The proportions of c18:2,t9,t11 found with Est-DFtol and AEst-EEhept were correlated to the dose of rpCLA (R = 0.87 and 0.51, respectively), whereas those found with Est-EEtol did not (R = 0.17). The Est- DFtol method is recommended because it minimizes the isomerisation of the polyunsaturated fatty acids and yields a more accurate measurement of the FAs profile

Milk-derived bioactive peptides exhibit antioxidant activity through the Keap1-Nrf2 signaling pathway

Abstract Bioactive peptides are relevant nutritional factors that exhibit many functions including antioxidant, antihypertensive, anticancer and antimicrobial properties. In this paper, four synthetic peptides ARHPHPHLSFM (A-11-M), AVPYPQR (A-7-R), NPYVPR (N-6-R) and KVLPVPEK (K-8-K) with sequences present in milk proteins were examined for their antioxidant properties. The compounds show moderate free radical scavenging activity in the ABTS and crocin assays (A-7-R and N-6-R) and lipid peroxidation inhibition in Caco-2 cells (N-6-R and K-8-K). All peptides, in particular K-8-K, activate the Keap1-Nrf2 system by allowing the translocation of the transcription factor Nrf2 from the cytosol to nucleus. This activation triggers the overexpression of the antioxidant enzymes Trx1, TrxR1, GR, NQO1 and SOD1. Furthermore, molecular modeling shows that K-8-K is able to hinder the interaction of Nrf2 with Keap1. The reported results show that the antioxidant action in cells of these bioactive peptides is mostly due to the activation of Keap1-Nrf2 signaling pathway

Loading of Polydimethylsiloxane with a Human ApoB-Derived Antimicrobial Peptide to Prevent Bacterial Infections

Background: medical device-induced infections affect millions of lives worldwide and innovative preventive strategies are urgently required. Antimicrobial peptides (AMPs) appear as ideal candidates to efficiently functionalize medical devices surfaces and prevent bacterial infections. In this scenario, here, we produced antimicrobial polydimethylsiloxane (PDMS) by loading this polymer with an antimicrobial peptide identified in human apolipoprotein B, r(P)ApoBLPro. Methods: once obtained loaded PDMS, its structure, anti-infective properties, ability to release the peptide, stability, and biocompatibility were evaluated by FTIR spectroscopy, water contact angle measurements, broth microdilution method, time-killing kinetic assays, quartz crystal microbalance analyses, MTT assays, and scanning electron microscopy analyses. Results: PDMS was loaded with r(P)ApoBLPro peptide which was found to be present not only in the bulk matrix of the polymer but also on its surface. ApoB-derived peptide was found to retain its antimicrobial properties once loaded into PDMS and the antimicrobial material was found to be stable upon storage at 4◦ C for a prolonged time interval. A gradual and significant release (70% of the total amount) of the peptide from PDMS was also demonstrated upon 400 min incubation and the antimicrobial material was found to be endowed with anti-adhesive properties and with the ability to prevent biofilm attachment. Furthermore, PDMS loaded with r(P)ApoBLPro peptide was found not to affect the viability of eukaryotic cells. Conclusions: an easy procedure to functionalize PDMS with r(P)ApoBLPro peptide has been here developed and the obtained functionalized material has been found to be stable, antimicrobial, and biocompatible

Host defense peptides identified in human apolipoprotein B as natural food bio-preservatives: Evaluation of their biosafety and digestibility

The employment of chemical agents in the food industry is raising several concerns by consumers and is leading to an increasing interest in natural food preservatives. Among alternatives, host defense peptides (HDPs) have attracted great interest for their ability to preserve food samples from contamination without altering their quality, taste, and organoleptic properties. Recently, we evaluated the applicability of ApoB-derived peptides as novel food bio-preservatives and demonstrated their ability to prevent chicken meat sample contamination when immobilized on chitosan films. To perform a further step towards the applicability of these peptides in the food field, here we evaluated peptides biosafety and digestibility. To do this, we used a multidisciplinary approach including the evaluation of the peptides' toxicity and antimicrobial activity, the analysis of resistance phenotype development, an in silico prediction of the peptides' susceptibility to proteases and the evaluation of the peptides' stability in simulated gastric and intestinal fluids. ApoB-derived peptides were found to be nontoxic when tested on human gastric carcinoma cells SNU-1 and on human colon-rectal adenocarcinoma cells HT-29, and not to induce resistance phenotype in Salmonella strains. Bioinformatic analyses showed that the peptides are susceptible to several proteases, as also confirmed by experiments in simulated gastric and intestinal fluids. Altogether these findings open interesting perspectives to the future applicability of ApoB-derived peptides as novel food biopreservatives

Energy Balance Estimated from Individual Measurements of Body Weight and Backfat Thickness of Heavy Pigs of Four Genetic Lines Fed Different Diets

Pigs of four genetic lines (GL): Anas (A), DanBred (D), Goland (G) and Topigs received either a conventional (140 g CP/kg and 46 g lysine/kg CP; C-CP) or a low protein diet (106 g CP/kg and 46 g lysine/kg CP; L-CP). Body weight (BW) and backfat depth (P2) were individually measured at the start and the end of two growing periods and individual feed intake (FI) was recorded daily. Body protein and lipid mass at the start and at the end of each period were estimated from BW and P2, and hence protein (Pr) and lipid (Lr) retention were computed. Energy requirement for maintenance (MEm), and growth (MEg) were estimated according to National Research Council guidelines, while ME intake (MEI) was computed from measured FI and ME content of the diets. The MEI/(MEm + MEg) ratio was used as index of efficiency. Differences among GL (P<0.001) were observed for Pr, which averaged 103, 113, 108 and 101 g/d for A, D, G and T, respectively, and Lr which averaged 204, 186, 194 and 172 g/d for A, D, G, and T, respectively. The L-CP diet reduced (P = 0.014) Pr by 8% compared to C-CP, but not Lr. Th e MEI/(MEm+MEg) index was influenced by GL (P<0.001) being 0.99, 0.96, 0.99 and 1.03 for A, D, G and T, respectively. Measurements of BW and P2 permits to achieve acceptable quantification of Pr and Lr. In this range of BW (90 to 165kg), gain composition is influenced more by GL than by the substantial reduction of CP and essential amino acids dietary density used in this trial

CK2 modulates adipocyte insulin-signaling and is up-regulated in human obesity

Insulin plays a major role in glucose metabolism and insulin-signaling defects are present in obesity and diabetes. CK2 is a pleiotropic protein kinase implicated in fundamental cellular pathways and abnormally elevated in tumors. Here we report that in human and murine adipocytes CK2-inhibition decreases the insulin-induced glucose-uptake by counteracting Akt-signaling and GLUT4-translocation to the plasma membrane. In mice CK2 acts on insulin-signaling in adipose tissue, liver and skeletal muscle and its acute inhibition impairs glucose tolerance. Notably, CK2 protein-level and activity are greatly up-regulated in white adipose tissue from ob/ob and db/db mice as well as from obese patients, regardless the severity of their insulin-resistance and the presence of pre-diabetes or overt type 2 diabetes. Weight loss obtained by both bariatric surgery or hypocaloric diet reverts CK2 hyper-activation to normal level. Our data suggest a central role of CK2 in insulin-sensitivity, glucose homeostasis and adipose tissue remodeling. CK2 up-regulation is identified as a hallmark of adipose tissue pathological expansion, suggesting a new potential therapeutic target for human obesity

Pathophysiological mechanisms and clinical evidence of relationship between Nonalcoholic fatty liver disease (NAFLD) and cardiovascular disease

Evidence suggests a close connection between Nonalcoholic Fatty Liver Disease (NAFLD) and increased cardiovascular (CV) risk. Several cross-sectional studies report that NAFLD is related to preclinical atherosclerotic damage, and to coronary, cerebral and peripheral vascular events. Similar results have been showed by prospective studies and also by meta-analyzes on observational studies. The pathophysiological mechanisms of NAFLD are related to insulin resistance, which causes a dysfunction in adipokine production, especially adiponectin, from adipose tissue. A proinflammatory state and an increase in oxidative stress, due to increased reacting oxygen species (ROS) formation with consequent oxidation of free fatty acids and increased de novo lipogenesis with accumulation of triglycerides, are observed. These mechanisms may have an impact on atherosclerotic plaque formation and progression, and they can lead to increased cardiovascular risk in subjects with NAFLD. This review extensively discusses and comments current and developing NAFLD therapies and their possible impact on cardiovascular outcome