Medial tunica degeneration of the ascending aortic wall is associated with specific microRNA changes in bicuspid aortic valve disease

Ascending aortic diameter is not an accurate parameter for surgical indication in patients with bicuspid aortic valve (BAV). Thus, the present study aimed to identify specific microRNAs (miRNAs/miRs) and their expression levels in aortic wall aneurysm associated with BAV according to severity of medial degeneration and to elucidate the association between the tissue expression levels of the miRNAs with their expression in plasma. Aortic wall and blood specimens were obtained from 38 patients: 12 controls and 26 patients with BAV with ascending aortic aneurysm. Of the patients with BAV, 19 had cusp fusions of right and left, 5 of right and non-coronary, and 2 of left and non-coronary. Two groups of patients were identified according to the grade of medial degeneration (MD): Low-grade D group (LGMD) and high-grade MD group (HGMD). Expression level of miR-122, miR-130, miR-718 and miR-486 were validated by reverse transcription-quantitative PCR in plasma and tissue samples. MD grade was found to be independent from the BAV phenotype. The HGD group showed increased expression levels of MMP-9 and MMP-2, and an increase in the number of apoptotic cells. Tissue expression levels of miR-718 and miR-122 were lower in the LGMD and HGD groups compared with expression in the control group; the HGD group showed increased levels of miR-486. Plasma expression levels of miR-122 were decreased in the LGMD and HGD groups, and miR-718 was only reduced in the HGD group. On the contrary, expression of miR-486 was increased in the LGMD and HGD groups. The data suggested that miR-486 may be considered as a non-invasive biomarker of aortic wall degeneration. Dysregulation of this putative biomarker may be associated with high risk of dissection and rupture in patients with BAV

Prognostic role of aldosterone in patients with acute coronary syndrome: short and medium term follow-up.

AIMS: Different studies have shown a correlation between aldosterone, atherosclerosis and ischemia in the past decade. Evidence exists for the relationship between high levels of aldosterone and augmented risk of cardiovascular diseases, such as hypertension, cardiac failure, coronary artery disease and stroke. The objective of this study was to determine the prognostic role of aldosterone in patients with myocardial infarction. METHODS:The study population included 96 consecutive patients admitted to our department for ST-elevated and non-ST-elevated myocardial infarction from June 2009 to March 2012. Plasma aldosterone levels were measured at admission to hospital in all patients. A 2-year prospective follow-up was performed, and fatal events and non-fatal events, such as reinfarction, congestive heart failure and arrhythmias, were recorded. RESULTS:Aldosterone levels at admission were associated with incidence of congestive heart failure (P\u200a=\u200a0.02), ventricular arrhythmias (P\u200a=\u200a0.01) and all complications (P\u200a=\u200a0.003) after 1-month follow-up. Moreover, high aldosterone levels gave important information in the medium term (24\u200a\ub1\u200a6 months). Specifically, aldosterone was a predictive variable of reinfarction (P\u200a<\u200a0.0001), congestive heart failure (P\u200a<\u200a0.0001) and adverse events (P\u200a=\u200a0.0002). The logistic regression analysis confirmed these results and showed that aldosterone may be predictive of adverse events at medium-term follow-up (odds ratio 1.1, 95% confidence interval 1.03-1.15, P\u200a=\u200a0.02). CONCLUSION:These data show a strong and significant correlation between plasma aldosterone levels at admission for myocardial infarction and fatal and nonfatal adverse events. Aldosterone appears to be a main marker of adverse clinical outcome, in accordance with the literature. These data suggest the need to identify whether antialdosteronic drug treatment, applied acutely in patients with aldosterone elevation, can influence favorably the prognosis of patients with myocardial infarction

Medium-term culture of normal human oral mucosa: a novel three-dimensional model to study the effectiveness of drugs administration.

Tissue-engineered oral mucosal equivalents have been developed for in vitro studies for a few years now. However, the usefulness of currently available models is still limited by many factors, mainly the lack of a physiological extracellular matrix (ECM) and the use of cell populations that do not reflect the properly differentiated cytotypes of the mucosa of the oral cavity. For this reason, we have developed a novel three-dimensional culture model reflecting the normal architecture of the human oral mucosa, with the main aim of creating a better in vitro model where to test cellular responses to drugs administration. This novel 3D cell culture model (3D outgrowth) was set up using an artificial extracellular matrix (MatrigelTM), allowing the interactions required for proper differentiation of the various citotypes which form the mucosal layer. Biopsies of human oral mucosa, in fragments of about 0.5 mm3, were placed onto 6.5mm Transwells, covered with MatrigelTM and grown in a specific culture medium. A gradual formation of an architectural structure similar to that of the in vivo oral mucosa was observed. Transmission electron and confocal microscopy were employed to characterize the newly developed model: the cell components (keratinocytes and fibroblasts) differentiated properly within the outgrowth and reconstituted, in vitro, the physiological structure of the human oral mucosa, including a stratified non-keratinized squamous layer composed of four different layers, a proper basal membrane and a lamina propria where fibroblasts produce ECM. Moreover, keratinocytes expressed CK5, CK13, CK19 and E-cadherin, whereas fibroblasts expressed collagen type I and IV, laminin and fibronectin. 3D outgrowths could be considered a valid alternative to animal models, and provide useful information for researchers interested in studying the responses of the human oral mucosa to locally delivered drugs or other exogenous treatments

Nandrolone decanoate interferes with testosterone biosynthesis altering blood-testis barrier components

The aim of this study was to investigate whether nandrolone decanoate (ND) use affects testosterone production and testicular morphology in a model of trained and sedentary mice. A group of mice underwent endurance training while another set led a sedentary lifestyle and were freely mobile within cages. All experimental groups were treated with either ND or peanut oil at different doses for 6 weeks. Testosterone serum levels were measured via liquid chromatography-mass spectrometry. Western blot analysis and quantitative real-time PCR were utilized to determine gene and protein expression levels of the primary enzymes implicated in testosterone biosynthesis and gene expression levels of the blood-testis barrier (BTB) components. Immunohistochemistry and immunofluorescence were conducted for testicular morphological evaluation. The study demonstrated that moderate to high doses of ND induced a diminished serum testosterone level and altered the expression level of the key steroidogenic enzymes involved in testosterone biosynthesis. At the morphological level, ND induced degradation of the BTB by targeting the tight junction protein-1 (TJP1). ND stimulation deregulated metalloproteinase-9, metalloproteinase-2 (MMP-2) and the tissue inhibitor of MMP-2. Moreover, ND administration resulted in a mislocalization of mucin-1. In conclusion, ND abuse induces a decline in testosterone production that is unable to regulate the internalization and redistribution of TJP1 and may induce the deregulation of other BTB constituents via the inhibition of MMP-2. ND may well be considered as both a potential inducer of male infertility and a potential risk factor to a low endogenous bioavailable testosterone

Medium-Term Culture of Primary Oral Squamous Cell Carcinoma in a Three-Dimensional Model: Effects on Cell Survival Following Topical 5-Fluororacile Delivery by Drug-Loaded Matrix Tablets

Since the activity of several conventional anticancer drugs is restricted by resistance mechanisms and dose-limiting side-effects, the design of formulations for local application on malignant lesions seems to be an efficient and promising drug delivery approach. In this study, the effect of locally applied 5-FU on cell death was evaluated both in a SCC4/HEK001 model and in a newly proposed 3D outgrowth model of oral squamous cell carcinoma (OSCC). Initially, the optimal drug dose was established by delivery of solutions containing different amounts of 5-FU. The solution containing 1% (w/v) of 5-FU resulted effective in inducing cell death with complete eradication of cell colonies. Buccal tablets were designed to deliver 5-FU locoregionally to the cancer lesions of the oral cavity. Tablets were prepared using a drug loaded matrix of acrylic/methacrylic acid copolymer containing 1% (w/w) of 5-FU and applied on 3D outgrowths. The drug release from tablets appeared to be sufficient to induce cell death as confirmed by transmission electron microscopy and enzymatic assay (TUNEL). After 120 h of treatment, when about 90% of the drug had been discharged from the tablets into the culture environment, 5-FU caused loss of cell-cell communications and apoptotic cell death. After 192 h, a complete disaggregation of the 3D oral outgrowths and the death of all the cells was observed. Buccal matrix tablets could be considered a promising new approach to the locoregional treatment of OSCC. Risks of systemic toxicity are avoided since very low drug doses are delivered

Il bipolarismo conflittuale. Il regime politico della Seconda Repubblica.

Si tratta di un contributo che affronta il tema della forma di governo. I due autori analizzano il funzionamento della forma di governo in considerazione del sistema politico bipolare evidenziando che nella esperienza più recente si è fortemente incrementata la conflittualità tra i vari attori politici

Decellularized Saphena: Biologic Scaffold for 3D Cellular Growth

Cardiovascular diseases are leading causes of mortality in Western society and countries adopting an Occidental Life style (1). Restoration of the circulation via bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and requires (vascular/tissue/circulatory/blood vessel - delete as appropriate) grafts. Normally the great saphena vein is used as (a/the) \u201cdonor\u201d blood vessel. It is collected from its superficial location on the leg and used in case of auto-transplantation in coronary artery bypass operations, and also for peripheral arterial bypass operations. Autologous great saphenous veins are often not available, and synthetic grafts have their limitations. Therefore novel techniques to produce alternative grafts have been developed, including tissue engineering which is a promising alternative to provide biocompatible grafts. Our aim is to reconstruct the endothelium layer of decellularized vein scaffolds using mesenchymal stem cells (MSCs) and growth factors. The use of allo/xenogenic material as a scaffold for donors has previosuly been embraced by the scientific community (2), and standardised procedures have been developed that consists of a decellularization phase of the donor element, a re-seeding phase of the decellularized scaffold and an implant phase on the host (3). In particular the opportunity to use a decellularized blood vessel as a scaffold for vascular tissue engineering, and the saphena vein specifically, is something well known (4-5). Our project concerns the development of blood vessel substitutes to be used mainly in arteriovenous fistulae surgery. We propose to develop a procedure that automates the initial phases (decellularization and re-seeding) to reduce the manipulation of the vein and to promote a more efficient and natural cell seeding. We have developed a specific bioreactor linked with a rotary infusion pump to create a closed circuit in which the donated saphena vein was integrated during the decellularization phase. We have enabled a flow of SDS (0.1% in sterile PBS) for 24h inside the circuit, and consequentially inside the saphena. After 24h the saphena was removed, decellularized from the circuit and samples were prepared for analysis. Observations were performed using an optical microscope and a transmission electric microscope to evaluate the condition of connective scaffold and the potential presence of cells. The total removal of cellular elements and the upkeep of the connective architecture, essential conditions for the subsequent seeding phase, were observed. Subsequently, we replaced the SDS solution inside the reservoir of the bioreactor with a specific stromal medium (alpha-MEM, supplemented with 20% FBS, 1% L-glutamine and 1% penicillin/streptomycin) containing adiposederived mesenchymal stem cells obtained from rats in order to obtain the re-seeding of the scaffold. Scaffolds were kept under these conditions for 7 days in order to obtain a proper vascular equivalent. After this period, sampleswere prepared for analysis. Observations with a light microscope confirmed the presence of properly differentiated endothelial-like cells that however were too few to form a continuous layer. In conclusion, our model represents a valid system to obtain natural scaffolds to be implemented in the creation of vascular equivalents, however, more experiments are needed in order to validate the results obtained so far. References 1. Diet Quality Scores and Prediction of All-Cause, Cardiovascular and Cancer Mortality in a Pan-European Cohort Study. Lassale C, Gunter MJ, Romaguera D, Peelen LM, Van der Schouw YT, Beulens JW, Freisling H, Muller DC, Ferrari P, Huybrechts I, Fagherazzi G, Boutron-Ruault MC, Affret A, Overvad K, Dahm CC, Olsen A, Roswall N, Tsilidis KK, Katzke VA, K\ufchn T, Buijsse B, Quir\uf3s JR, S\ue1nchez-Cantalejo E, Etxezarreta N, Huerta JM, Barricarte A, Bonet C, Khaw KT, Key TJ, Trichopoulou A, Bamia C, Lagiou P, Palli D, Agnoli C, Tumino R, Fasanelli F, Panico S, Bueno-de-Mesquita HB, Boer JM, Sonestedt E, Nilsson LM, Renstr\uf6m F, Weiderpass E, Skeie G, Lund E, Moons KG, Riboli E, Tzoulaki I. PLoS One 2016;11:e0159025. doi: 10.1371/journal.pone.0159025. eCollection 2016. 2. The use of xenogeneic small intestinal submucosa as a biomaterial for Achilles tendon repair in a dog model. Badylak SF1, Tullius R, Kokini K, Shelbourne KD, Klootwyk T, Voytik SL, Kraine MR, Simmons C. J Biomed Mater Res. 1995 Aug;29(8):977-85. 3. Enhanced in vivo function of bioartificial lungs in rats. Song JJ, Kim SS, Liu Z, Madsen JC, Mathisen DJ, Vacanti JP, Ott HC. Ann Thorac Surg. 2011 Sep;92(3):998-1005; discussion 1005-6. doi:10.1016/j.athoracsur.2011.05.018 4. Decellularized vein as a potential scaffold for vascular tissue engineering. Schaner, Patrick J et al. Journal of Vascular Surgery, Volume 40 , Issue 1 , 146 - 153 5. Reconstruction of small diameter arteries using decellularized vascular scaffolds. Nagaoka Y, Yamada H, Kimura T, Kishida A, Fujisato T, Takakuda K. J Med Dent Sci. 2014 Mar 19;61(1):33-40

Indoor environmental quality survey: a brief comparison between different Post Occupancy Evaluation methods

Building occupants are important factor in giving information on indoor conditions such as comfort, productivity, building performance and occupants\ub4 health. Even that, being their appraisal mainly based on subjective judgments, it is difficult to set up a standard and objective method for this purpose. The present work aimed to describe, analyze and compare the most common tests on this topic, in order to highlight critical, failings and strengths

LA VALORIZZAZIONE DELLA BIOMASSA ATTRAVERSO IL RICICLAGGIO DEI RIFIUTI DELLA FILIERA OLIVICOLO OLEARIA

IL PRESENTE LAVORO DEFINISCE STRATEGIE PER LA VALORIZZAZIONE DELLA BIOMASSA ATTRAVERSO IL RICICLAGGIO DEI RIFIUTI DELLA FILIERA OLIVICOLO OLEARIA