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Opis: Objectives: We have shown that age and pathology impair the biological properties of stem cells isolated from human hearts (CSC) and functional assays showed differences between CSC isolated from normal (DCSC) and end-stage failing (ECSC) hearts. As alterations of mechanical properties of the myocardium, such as stiffening and increased wall stress, are crucial features of cardiac remodeling, this work addresses the biological effects exerted on CSC by mechanical stimuli. Materials and methods: DCSC and ECSC were cultured under defined conditions to mimic specific features of the pathologic condition: increased mechanical loading (up to 15%, cyclic at 1 Hz), differential substrate stiffness (ranging from 1 to 231 kPa), differential cell densities. After 24, 48 and 72 h, cells were fixed and stained for analysis of proliferation and subcellular localization of YAP or lysed for RT-PCR analysis.Results: Cyclic stretch was significantly associated with both increased proliferation of DCSC (n = 6, p<0.0001) and ECSC (n = 4, p = 0.003), and with a significant reduction of nuclear localized YAP (nYAP) as a function of time (p<0.05). However, while significant correlation between cell density and decreased nYAP (p = 0.003, r2 = 0.37) characterized ECSC, this was not evident for unstretched DCSC, suggesting a less stringent regulation of contact inhibition in DCSC. These data were further confirmed by seeding cells at differential density. As opposed to what previously shown for epithelial cell lines, DCSC did not reduce nYAP positivity as a function of cell density, when grown in serum containing medium, suggesting that soluble factors present in the serum could maintain the nuclear localization of YAP, independently from the cell density. In line, serum significantly increased the nYAP expressing cells in DCSC, while a significant positive correlation between cell density and nYAP positivity can be demonstrated in DCSC cultured in serum free medium. RT-PCR for YAP-regulated targets confirmed immuno- fluorescence data. Furthermore, independently from the pathologic status, cyclic stretch was significantly associated with a persistent YAP signaling at high cell density. Besides, tension and assembly of cytoskeletal network, induced by increasing substrate stiffness, correlates with nYAP (p<0.05) and YAP transcriptional activation (p<0.05). Conclusions: D- and ECSC differ in their mechanosensing properties. However, in the first cell type, nYAP localization is dictated by the combined action of paracrine factors and cytoskeletal tension, thus reducing the contact inhibition effect. This finding is in line with a more primitive phenotype of SC isolated from normal hearts.
Ključne besede: Stem cells, human cardiac stem cells, mechanosensing, mechanotransduction, heart failure, YAP
Objavljeno v RUNG: 21.03.2017; Ogledov: 4342; Prenosov: 0
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Opis: Today it is widely acknowledged by international institutions, national and local administrations, and the civic society at large, that the whole of Cultural Heritage (CH), in its tangible and intangible forms, is part of our individual and collective memory, a marker of cultural identity and at the same time a demonstration of social heterogeneity and complexity. This has led to a progressive recognition of the importance of not only protecting, but also valorising CH, through its promotion and the facilitation of its fruition. Benefiting from CH depends on what and how is done to promote it and also to create a competitive advantage for the ‘cultural economy’ which is based on its reproduction. Tourism is an industry which uses CH and to some extent achieves the objective of its valorisation, widening the market for its fruition and the significance of heritage for different ‘publics’, but only to the extent that it is sustainable in its widest sense. This sets a number of challenges regarding how the heritage is offered, explained, and made accessible to visitors. In this sense, the valorisation of CH is at the core of value generation in tourism, and the use of ICTs (Information and Communication Technologies) is the channel through which increasingly cultural resources are experienced in competitive and sustainable way. New ICT tools restructure the relationship between image and word, leading to new ways to imagine and interpret CH, and offer great potential for value creation in tourism whereby perception and visual processing are essential elements of communication and experience. Interpretation, which can be informative, inspirational and entertaining, spurs understanding and appreciation, and ultimately engages visitors in CH. Besides, online social networks have a central place in the shaping of experience of the contemporary visitor and offer potential for a smart marketing approach. This Master thesis aims to demonstrate that managing the flows of information through the new technologies may indeed enhance CH valorisation, and that the introduction of digital devices and virtual technologies should follow a visitor-centred approach. It also intends to acknowledge that the application of ICTs can have a significant return in terms of economic income, stimulating and generating a stronger understanding among cultural and public institutions and organizations. The first section of this work reviews the relationship between CH and its social dimension, presenting CH as a lever to enhance social and economic development; it presents ICT and their relation with cultural tourism, illustrating the ongoing transformation of visitors into active actors in the valorisation of CH. The second section presents two case studies of digital devices applied to CH valorisation in different contexts; the impacts of the two projects in relation to heritage itself, its users and the territory are carefully analised. The research represents a starting point for more work encased in contemporary studies on the management of CH, as well as in the developing area of digital interpretation, towards a more general development of a model of sustainable CH tourism.
Ključne besede: CH management, cultural tourism, ICT, interpretation, CH experience
Objavljeno v RUNG: 17.03.2017; Ogledov: 5177; Prenosov: 229
Celotno besedilo (2,87 MB)

Opis: Short peptides can be designed in silico and synthesized through automated techniques, making them advantageous and versatile protein binders. A number of docking-based algorithms allow for a computational screening of peptides as binders. Here we developed ex-novo peptides targeting the maltose site of the Maltose Binding Protein, the prototypical system for the study of protein ligand recognition. We used a Monte Carlo based protocol, to computationally evolve a set of octapeptides starting from a polialanine sequence. We screened in silico the candidate peptides and characterized their binding abilities by surface plasmon resonance, fluorescence and electrospray ionization mass spectrometry assays. These experiments showed the designed binders to recognize their target with micromolar affinity. We finally discuss the obtained results in the light of further improvement in the ex-novo optimization of peptide based binders.
Ključne besede: peptides, docking, optimisation, computation, maltose binding protein, probe, ligand
Objavljeno v RUNG: 12.10.2016; Ogledov: 4009; Prenosov: 140
Celotno besedilo (4,27 MB)

Ključne besede: microscopy, imaging systems, thermal lensing, medical imaging, biological imaging, cell analysis
Objavljeno v RUNG: 17.05.2016; Ogledov: 4453; Prenosov: 143
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