Zebrafish (Danio rerio) have the remarkable ability to regenerate the heart, by a process referred to as epimorphic regeneration, the regrowth of amputated structures from an anatomical complex stump. Networks of transcription factors regulate heart development, maintenance, and regeneration in a dose-dependent manner, but the effects of translational regulation on the titration of these pathways are largely unknown. Here, with In Situ Hybridization (ISH) experiments, it was tested in regenerating hearts the presence and the timing expression of genes involved in FGF regulation pathway. In-vivo experiments showed that the response to the injury started already at 3 hours post amputation (hpa), just around the surgered site, by the reactivation of developmental genes such as dusp6 (mkp3), erm, pea3, raldh2, and sef. Between 6 and 12 hpa all the amputated hearts evidenced the expression of other genes such as etv5 and sprouty4, showing the reactivation of development genes. With the aim to optimize the media to reproduce the regeneration process, it was also tested the ability of zebrafish heart to survive in ex-vivo cultures after the amputation of ventricular apex. Injured hearts cultured with BCI, FGF-2, Thrombin, and with a cocktail of FGFs were able to maintain expression of raldh2 between 7 and 14 days post amputation (dpa), whereas PDGF and Basement Membrane Extract (BME) supplemented with FGFs cocktail allowed the raldh2 expression until 30 dpa. Regenerating hearts in ex-vivo conditions were able to survive and make contractions, and surprisingly showed different degree of cell replication in dependence to the culture media. The highest level of duplication, detected by BrdU incorporation, was observed at 14 dpa in hearts cultured with addiction of BCI. However operated hearts in-vivo showed a complete regeneration after 30 dpa whereas ex-vivo cultured the hearts displayed only a partial regeneration. Moreover, in ex-vivo, a big diminution of the clot site was observed in hearts cultured with BCI, suggesting that this compound could interact with FGF pathway, stimulating and supporting the heart regeneration process.
Furthermore, Nakamura (Nakamura et al.,1998) showed that, in patients subjected to bypass surgery, Trx inactivation was deleterious in I/R injury. To date, there are no indications about the role exerted by endogenous or exogenous SelT in cardioprotection. In the present study, we found that pharmacological post-conditioning with the full length rSelT and PSELT, which encompasses SelT active site, protects the Langendorff perfused rat hearts exposed to I/R injury. We found that, compared to the IS detected in hearts exposed to I/R (~75%), hearts post-conditioned with PSELT showed a remarkably reduced IS (~40%), which was similar to that detected in hearts perfused with full length rSelT (~35%). IS reduction correlates with systolic function recovery and with the absence of contracture development. These effects raise the question of the site of action of PSELT. Although no definitive answer can be provided at this point for cardiomyocytes, we have recently shown using neuroblastoma cells that a fluorescent PSELT crosses the plasma membrane and may thus act intracellularly (manuscript in preparation). Whether PSELT acts at the level of the sarcoplasmic reticulum where SelT is localized remains to be determined. Cardioprotection was more evident in ischemic hearts exposed to PSELT at the EC50 concentration (5 nM) than in hearts exposed to the highest concentration of the peptide (100 nM). This is shown by the better post-ischemic recovery of dLVP observed in hearts exposed to 5 nM of SelT. In both cases, PSELT reduced contracture. The striking protection elicited by the peptide is indicated by the significant IS reduction, observed with both PSELT concentrations tested (5 and 100 nM). This specific concentration-dependent behaviour resembles that described for major exogenous antioxidant agents known to induce protection at lower doses, but not at higher doses (Bouayed and Bohn, 2010). In fact, in vitro and in vivo evidence indicates that high concentrations of antioxidants may be harmful, due to pro-oxidative effects and/or their reactivity with the physiological ROS concentrations required for optimal cellular functioning. This results in a redox disequilibrium in favour of oxidation (Bouayed
Methylglyoxal (MGO), a highly reactive dicarbonyl compound formed as by-product of glycolysis, is an ubiquitous metabolite of cellular metabolism. Therefore, it is produced in all cells, both under normal and pathological conditions. Under physiological circumstances, MGO is detoxified through the glyoxalase system, of which Glyoxalase 1 (Glo1) is the rate limiting enzyme. In pathological conditions, as in chronic hyperglycemia, high blood glucose levels lead to increased MGO accumulation. It is known that MGO plays a major role in endothelial cell damage and development of vascular disease. We have previously demonstrated that MGO induces endothelial insulin resistance both in vitro and in animal models. In the last few years, many evidence has provided a link between microRNAs (miRNAs) and diabetic complications. Indeed, miRNAs regulate cellular molecular pathways, including insulin signaling, thus controlling the pathophysiology of vascular bed. This study includes the investigation of two aspects of MGO effects on the pathophysiology of diabetes mellitus (DM) and its associated complications: 1. the evaluation of MGO accumulation on glucose homeostasis and vascular function in a mouse model knockdown for Glo1 (Glo1 KD ) and 2. the analysis of miRNAs contribution in MGO induced damaging effect on insulin responsiveness in mouse aortic endothelial cells (MAECs).
Here, we developed an isogenic cell model of “stemness” to facilitate protein biomarker discovery in breast cancer. For this purpose, we used knowledge gained previously from the study of the mouse mammary tumor virus (MMTV). MMTV initiates mammary tumorigenesis in mice by promoter insertion adjacent to two main integration sites, namely Int-1 (Wnt1) and Int-2 (Fgf3), which ultimately activates Wnt/β-catenin signaling, driving the propagation of mammary cancer stem cells (CSCs). Thus, to develop a humanized model of MMTV signaling, we over- expressed WNT1 and FGF3 in MCF7 cells, an ER(+) human breast cancer cell line. We then validated that MCF7 cells over-expressing both WNT1 and FGF3 show a 3.5- fold increase in mammosphere formation, and that conditioned media from these cells is also sufficient to promote stem cell activity in untransfected parental MCF7 and T47D cells, as WNT1 and FGF3 are secreted factors. Proteomic analysis of this model system revealed the induction of i) EMT markers, ii) mitochondrial proteins, iii) glycolytic enzymes and iv) protein synthesis machinery, consistent with an anabolic CSC phenotype. MitoTracker staining validated the expected WNT1/FGF3-induced increase in mitochondrial mass and activity, which presumably reflects increased mitochondrial biogenesis. Importantly, many of the proteins that were up-regulated by WNT/FGF-signaling in MCF7 cells, were also transcriptionally over-expressed in human breast cancer cells in vivo, based on the bioinformatic analysis of public gene expression datasets of laser-captured patient samples. As such, this isogenic cell model should accelerate the discovery of new biomarkers to predict clinical outcome in breast cancer, facilitating the development of personalized medicine.
Goal of this research project is to explore the effects that increased levels of GCs exert in brain regions particularly susceptible to stressful stimuli, such as hippocampus, in DMD. The animal model used has been the mdx mouse, a genetic model of the disease in which a spontaneous point mutation in the exon 23 induces an anticipated stop codon in the dystrophin gene, resulting in absence of full-length dystrophin synthesis. Numerous experimental evidences are shedding a light on the role that Dp427 plays in different brain regions. Among these are hippocampus and cerebellum, which are field of demonstrated brain physiological failures and neurological disorders in both DMD patients and animal models. These areas are in part overlapping with those recognized as major targets of GCs, which are released at high levels in response to both emotional and physical stressors. This makes imperative to better investigate whether the mode of action of GCs in the brain of wild type and dystrophic mice is similar, or whether important differences related to dysfunction of the DGC complex, determined by lack of Dp427, can be revealed. Since DMD patients are subjected to repeated and prolonged application of corticosteroids, directed at lowering recurrent muscular inflammatory events, it is of some importance identifying factors, which could aggravate the already compromised neurological conditions of young DMD patients.
Many mathematical models of the cardiovascular system have been published since Grodins made the first global dynamic model of one in 1959 (Grodins, 1959). Guyton’s system analysis of circulatory regulation (Guyton et al., 1972) was performed with a complex model which comprised of 354 blocks, each representing one or more mathematical equations describing some physiological facet of circulatory function. In general, each of the functional blocks has been the subject of research investigation by one or many investigators. Some other modeling studies dedicated to characterizing certain causal relationships between circulatory variables and hemodynamic consequences using mathematical models were conducted by (Beyar et al., 1987, Maughan et al., 1987, Santamore and Burkhoff, 1991). Based on these works, a significant improvement was made by (Sun et al., 1997) who constructed an elaborated right–left heart interaction mathematical model capable of predicting cardiac hemodynamics for not only normal but also various pathological conditions. For ventricular modeling a considerable step forward was done by (Campbell et al., 1982a, Campbell et al., 1982b) that developed the so- called time-varying elastance model, as proposed by (Sunagawa and Sagawa, 1982). More recent works integrate models at different scales, e.g. from cells to system (Shim et al., 2006) or computational fluid dynamics models with lumped parameter models (Liang et al., 2007).
KMT2D were found in more than 50% of patients with KABUK1, with the majority of mutations resulting in the premature termination of the protein product. In addition, mutations in KDM6A, an H3K27me demethylase gene, were also reported to contribute to less than 10% of this syndrome, and this type is referred as Kabuki syndrome 2 (KABUK2; OMIM 300867). Recently, Bögershausen et al. identified two mutations in RAP1A/B, which encode the Ras family small GTPases, in patients with KABUK1 by whole exome sequencing. The authors also demonstrated that mutant RAP1 morphant phenocopied KDM6A and KMT2D mutants in zebrafish, and that the MEK/ERK pathway signaling was perturbed in RAP1- and KMT2D-defective cells. Interestingly, these phenotypes were rescued by treatment with an MEK inhibitor. On the other hands, the reduction in neurogenesis and hippocampal memory defects exhibited in a KABUK1 mouse model were ameliorated by the treatment with a histone deacetylase (HDAC) inhibitor, AR-42. Furthermore, a ketogenic diet rescued hippocampal memory defects through the elevation of beta-hydroxybutyrate, an endogenous HDAC inhibitor, in the same mice model. Taken together, these results potentially provide diverse therapeutic directions to treat, or at least mitigate, the symptoms of KABUK1.(Kim, Lee et al. 2017)
Different substances were selected thorough the years of study, among those NPS with structures and action similar to other compounds already prohibited in sport competitions. The substances of interest were selected according to University of Ferrara department of morphology, surgery and experimental medicine, section of legal medicine, a collaborative centre of Italian Early Warning System (IEWS) and University Cattolica of Rome. This collaboration led to the establishment of a multicentric collaborative group for the IEWS. The aim was to introduce these selected NPS as recognized doping agents searched by laboratories routine drug test of World Antidoping Agency (WADA), or forensic toxicology laboratories. With this aim specific compounds were selected and their potential effects were evaluated through in vivo behavioural studies employing murine model as a model of human behaviour and metabolism. The substances were administered to mice groups and behavioral studies were carried out at University of Ferrara to establish potential stimulant effects, which configure substances as stimulant compound in- competition. When a substance has showed typical effects as a doping agent, metabolism studies were carried out by our laboratory employing human liver microsomes or CYPs isoform as a model of human oxidative metabolism, and samples were analysed through liquid chromatography mass spectrometry techniques.
fibres in the tissue, challenging the cellular homeostasis. As a result, the macromolecular assembly of collagen and elastin in the skin may be impaired or modified, thus causing the skin to become thinner and lose elasticity . The process leads to the catabolism of extracellular matrix components (collagens and proteoglycans) with a loss of matrix . This phenomenon is particularly due to matrix metalloproteinases expression. A strong degradation of type I collagen also occurs in irradiated skin compared with nonirradiated skin . In addition, excessive exposure induces alteration of the biomechanical properties of dermal connective tissue resulting in dryness and reduced skin elasticity . These changes are coupled with DNA damage induced by reactive- oxygen species (ROS) production  that not only lead to premature aging of the skin but also increase the risk of contracting skin cancer. Moreover, ROS, inducing apoptosis, reduces the number of skin fibroblasts and decreases their regenerative capacity, leading to increased skin sagging . Furthermore, environmental pollutants may contribute to prompt symptoms of extrinsic skin aging, including coarse wrinkles, irregular pigment spots, and elastosis . Thus, the identification of new compounds, which are effective in protecting skin cells, is an important tool to prevent and/or to contrast the damage induced by ultraviolet radiation. Currently, there is a great tendency to use different skin care product formulations, considering their effect on the reduction of free radicals production generated from ultra- violet radiation. To date, stimulation of skin biorejuvenation is facilitated by minimally invasive intradermal injections of biologically active substances [15, 16] such as hyaluronic acid- (HA-) based gels and dermal fillers  alone or in combination with other molecules [18, 19]. Reports show that HA accelerates in vitro processes related to wound healing  and in vivo tissue
In the second part of the study, we explored post mortem pathology-MRI correlates and specifically focused on an advanced MRI technique (magnetization transfer ratio -MTR-), ideally detecting myelin content. MTR is widely used in MS observational studies and clinical trials, but its pathological correlates remain unclear. MTR maps were acquired at 3 Tesla from sixteen fixed MS brains and four healthy controls. 101 tissue blocks were immunostained and quantified, as previously described. After semi-automatic registration of digitized histologic sections and MTR maps, ROIs were manually defined. Associations between MTR and each stain were explored using linear mixed regression models (with cassettes nested within patients); differences in the associations between ROIs were explored using interaction terms. Lower MTR was associated with lower levels of NF200, SMI94, CD68, IBA1 and GFAP, with higher levels of CD8 and greater mitochondrial damage; MTR was more strongly associated with SMI94 in GM than WM. In a multivariate linear mixed regression model including all ROIs and brains, SMI94 was the best correlate of MTR. Myelin immunostain intensity is the strongest correlate of MTR, especially when measured in the GM. However, the additional histological correlates of MTR have to be kept in mind when interpreting the results of MTR clinical studies and designing experimental trials in MS.
A cross comparison of the Belgian-Hungarian results with respect to the three Italian regions estimates has demonstrated that the estimation for Belgium presents the same scale value as the Italian one, while the Hungarian results are more distant. This comparison does not constitute a method for checking the estimation goodness, but rather, given the lack of observed accounting costs, provides a narrow judgement of the estimate scale and the degree of approximation to the three Italian regions validated estimates. It is quite clear that, for a deeper validation of the results achieved for Belgium and Hungary, observed accounting costs per crop are necessary. To date, the only information available for this scope is the estimation of soft wheat developed within WP5, which has reached estimates very close to the values obtained for the Italian and Belgian case studies; while, for Hungary the PMP estimates are much higher than the WP5 outcome.
To conclude, lots of researchers stated that strategic leadership does indeed matter in organizations (Cannella & Monroe, 1997)(Thomas, 1988). However the real question should not be whether the strategic leadership matters or not, but rather under which conditions, when, how and on what criteria (Boal & Hooijberg, 1980). According to the upper echelon theory (Hambrick & Mason, 1984) the organization is WKHUHIOHFWLRQRIWKHOHDGHUV¶YDOXHVDQGFRJQLWLRQVDQGWKHDPRXQWRIGLVFUHWLRQZLOO moderate the relationship between strategic decisions and work outcomes (Kaplan & Kaiser, 2006) :LNLSHGLDGHILQHGGLVFUHWLRQDV³The ability to make decisions which represent a responsible choice and for which an understanding of what is lawful, right or wise may be presupposed ´DQGIURPDPRUHSUDFWLFDOSRLQWRIYLHZLW reflects the degree to which managers can turn their intentions into reality (Kaplan & Kaiser, 2006). Discretion summarizes three kinds of factors: environmental constraints, individual differences and organizational factors, moreover it is a reflection of demographic and personality characteristics (Cannella & Monroe, 1997). When it is high, leaders are relatively free to do as they wish and if the contrary, judgments and behaviors are constrained. Since leaders and organizational outcomes are linked by discretion, a dilemma is posed: without discretion, the leader is unable to influence firm performance instead with discretion could put self-interest ahead of their responsibilities and obligations (Kaplan & Kaiser, 2006). In regard to this topic the discussion is still open.
The third dataset consists in displacement time series derived from Sentinel-1A DInSAR data, using the SBAS technique (Fiaschi et al., 2018). The data consist of 54 (track 95) SAR images, acquired in C-band (5.6 cm of wavelength) with vertical co-polarization and in descending geometry. The SAR images were acquired with the Interferometric Wide swath mode from 17 November 2014 to 17 May 2017 and have a revisit time of 12 days, a look angle of 39.0° and a ground resolution of about 15 m × 15 m by adopting a multi-look of 4–1 in range and azimuth directions (Fiaschi et al., 2018). The processing workflow is structured in progressive steps: generation of connection graph, interferograms generation, phase unwrapping, refinement, first estimation of velocities, final estimation of velocity and atmospheric contribution removal, displacement time series estimation. The final outputs are imported in a Geographic Information System (GIS) for being classified and interpreted. During the processing a temporal baseline of 110 days and a perpendicular baseline of 2% have been chosen. The satellite’s orbit geometry was estimated using precise orbits data available from European Space Agency (ESA), while topographic phase residuals were removed using the Shuttle Radar Topographic Mission (SRTM) DEM with a ground resolution of around 30 m × 30 m (pixel size) (Fiaschi et al., 2018). The obtained DInSAR results are not calibrated but compared and validated with in-situ observations provided by previous high-precision levelling surveys and GPS data (TGPO and PTO1 European dataset) (Fiaschi et al., 2018).
phases, related to (i) identifying abbreviations within a source text, (ii) resolving the identified abbreviations by finding their corresponding ex- planation, and (iii) matching these explanations with entities of a de- sired domain, given a domain entity repository. The overall approach overcomes typical drawbacks of similar techniques, by employing: light- weight morphological checks and criteria for the identification and resolu- tion phases, with minimal Natural Language Processing (NLP) overhead, resulting in extremely fast execution times; weak constraints upon the structure of the candidate words, in order to broaden recall with respect to other methods using strong constraints; full-text papers as specific targets around which the whole system has been designed and built, and which differ from their abstract counterparts not just in terms of length, for they feature a far higher level of complexity and “unstructuredness” and a wider range of abbreviation forms, especially in chaotic domains like biomedical publications; a final entity recognition phase, which is indeed lacking in the major abbreviation discovery approaches.
The exposure of perfused working heart preparations of eel, goldfish and frog to increasing concentrations of either Serp or pGlu-Serp showed that these CgA C-terminal fragments induced a significant cardio-suppressive influence. We also found that the inhibitory effect on contractility was more evident in the presence of pGlu-Serp than Serp. In fact, in all species tested, pGlu-Serp caused a concentration-dependent reduction of contractile perfor- mance, significant from the concentration of 33 nM, whereas Serp was effective only in the frog heart, in which it reduced SV starting from a higher concentration (69 nM). The magnitude of Serpinin effects (changes of approximately 10–15%) is in the same range of that reported for the CgA-derived fragments VS-1 and CST (see for example, Imbrogno et al., 2004, 2010 for eel; Mazza et al., 2007, 2008 for frog), the full length CgA ( Pasqua et al., 2013 ), and other cardioactive agonists (e.g. nesfatin-1: Mazza et al., 2015a,b ; Isoproterenol: Tota et al., 2004 ), whose biological significance in the modulation of cardiac performance is well established. In par- ticular, we previously showed in the teleost A. anguilla ( Imbrogno et al., 2004, 2010 ) and in the frog R. esculenta ( Corti et al., 2004; Mazza et al., 2008 ), that the peptides VS-1 and CST exert dose- dependent negative contractile effects which started at 33 nM (#15%), reaching a maximum (by #20%) at the highest concentra- tion tested. On both species, the two peptides are also able to coun- teract the influence of physiological cardio-stimulating factors (e.g. catecholamines and Endothelin-1) ( Imbrogno et al., 2004, 2010; Corti et al., 2004; Mazza et al., 2008 ). This is in agreement with the idea that CgA generates different hormone peptides which work as homeostatic stabilizers of a number of physiological pro- cesses, as illustrated for example by Koeslag and co-workers (1999) . This would be of physiological importance under condi- tions of excessive adrenergic stimulations, such as under stress conditions in which CgA peptides counterbalance the effects of Iso- proterenol and Endothelin 1 (see Tota et al., 2004; Mazza et al., 2008 , for references). Interestingly, on the isolated and Langendorff perfused rat heart, the exposure to similar concentrations of pGlu- Serp induced a dose-dependent increase of contractility and relax- ation ( Tota et al., 2012 ). The reason for this divergence in the cardiac effects of Serpinin peptides in mammalian vs non-mammalian vertebrates is unknown. However, this is not surprising since the response to cardio-active agents is strongly influenced by species-specificity. For example, the non-specific b-AR agonist Iso- proterenol is known to induce a positive inotropic and chronotro-
To this extent, after describing the hydraulic properties of volcanic rocks, the analysis of water cir- culation was performed through the observation of the reconstructed water table from 2009 field data in comparison with previously acquired piezometric maps. Starting from the ancient volcanic deposits emplaced in the eastern part (TGT, 1 in Fig. 2.18). The paroxysmal explosive eruption that produced the emplacement of the “tufo rosso a scorie nere” Sabatino (TRSNS, 2 in Fig. 2.18), changes strongly the paleo-morphology of this sector (as well as the eastern). Pyroclastic deposits 10 to 25 m thick were deposited. These deposits of TRSNS, having predominant pozzolana charac- teristic, highly permeable in depressed paleo-morphology, were transformed, with cooling and deposition of zeolites in tuff lithoid reddish, fractured permeability and modest porosity. For its permeability in pozzolana facies, its thickness, even higher than 20 meters, and extension across the area, TRSNS could be considered one of the main aquifer in Sabatini context. The unity of TRSN Sabatino emerges fairly continuous at the edge of the volcanic district. This unit is considered a good stratigraphic indicator: its distribution must have been influenced by topographical conditions existing prior to its emplacement and not present in the high structural of Baccano-Cesano (Cioni, 1993), still morphologically prominent at the time of deposition (De Rita et al., 1993).
However, the MSSM in turn can be formulated as the low energy description of an high energy theory less rigid than the mSUGRA models. Without assuming any special scenario for the ultimate theory, the choice of appropriate experimental quantities to compare to the experimental predictions becomes of particular concern. In this context, the dynamics of flavour physics in the quark sector provides a promising way to explore the structure of the SUSY theories. We are thinking, in particular, to the already mentioned FCNC processes. The theoretical predictions for these observables, receiving loop-induced contributions from the new SUSY particles, must be as general as possible. The philosophy under this statement is clear: whatever the ultimate theory, once the SUSY parameter space is provided at a scale around the EW one, it is possible to directly compute the theoretical prediction for these processes, compare with the experimental number and decide if the model is phenomenologically aviable.
antimicrobial activities and activate neutrophils, with important implications in inflammation and innate immunity. Staphylococcus aureus is an opportunistic pathogen and the leading cause of a wide range of severe clinical infections and one of the most important cause of hospital-acquired infections, in fact infections caused by this bacterium have classically an important impact in morbidity and mortality in the nosocomial and community scene. Furthermore, this pathogen is the primary cause of surgical site infections and the most frequently isolated pathogen in Gram-positive sepsis. In the specific field of cardiovascular disease S. aureus leading infective cause of destruction of endocardial tissue after implantation of prosthetic heart valve. This pathogen is also notorious for its ability to resist the available antibiotics and dissemination of various multidrug-resistant S. aureus clones that limit therapeutic options for a S. aureus infection. Aslam et al. in 2013 shown that Ctl is resistant to the degradation of S. aureus protease and is the most antibacterial CgA derived peptide against this bacterium. The aim of study was to evaluate the: 1) Effects of Chr on isolated and Langendorff perfused rat hearts in basal and pathological conditions; 2) In vitro antibacterial activity of a synthetic Cateslytin-derived peptide to cover artificial heart valves and prevent infection by S.