1,805 research outputs found
A novel Yap/Taz zebrafish reporter reveals a role of Hippo pathway transducers in angiogenesis
No full textYAP and TAZ, by orchestrating cell proliferation, cell death and cell-fate decisions, are key players of a complex network of signaling pathways acting during development. Deregulation of YAP/TAZ signaling causes robust organ overgrowth during organogenesis, which translates to loss of tissue homeostasis in the adult and consequent cancer development. YAP/TAZ are transcriptional co-activators that interact with TEAD transcription factors to promote cell proliferation and survival. Their transcriptional activity is regulated by nucleocytoplasmic shuttling and nuclear accumulation, which are controlled by the Hippo kinase cascade, but also by mechanical cues sensed by the cell and by other pathways. Among these, Wnt/β-catenin takes on a particular relevance, since it was recently shown to regulate YAP/TAZ activity through AXIN-mediated sequestration of YAP/TAZ in the β-catenin destruction complex.
Here, we describe the generation, validation and characterization of a novel biosensor zebrafish reporting the activity of Yap/Taz. It expresses nuclear mCherry, eGFP or the destabilized green fluorescent protein VenusPEST under the control of a promoter fragment of the human YAP/TAZ target gene CTGF, that contains 3 TEAD DNA-binding sites. Several independent founder fish transmitting the transgene to the germline were identified and used to establish the stable reporter lines. All stable transgenic fish shared a similar expression pattern, which was maintained in subsequent generations. Knockdown and overexpression approaches were used to validate the reporter. Co-injection of two morpholinos targeting Yap and Taz pre-mRNAs reduced the reporter signal, whereas injection of mRNAs coding for a constitutively active form of Yap, Taz and Tead (YAP-5SA, TAZ-4SA, TEAD-VP16) increased it. The CTGF-based transgenic lines represent therefore bona fide Yap/Taz reporters.
During development, strong reporter signal is visible mainly in the lens and otic vesicles, the pharyngeal arches, the heart, the pectoral fin and the vasculature, but the reporter protein expression is also detected in many other tissues and organs. The almost ubiquitous activation of Yap/Taz observed during early embryogenesis, consistent with the general role of YAP/TAZ in promoting cell proliferation and organ growth, is largely silenced in the adult fish, where the reporter signal is restricted to the lens, the ovary, the heart and the whole vasculature. We also showed that the CTGF-based biosensor zebrafish is able to report Yap/Taz activation during larval and adult fin regeneration, as expected from the role that YAP/TAZ signaling plays in the regenerative processes.
The zebrafish CTGF-based reporter permitted to show in a living organism during development the regulation that the Wnt/β-catenin pathway exerts on Yap/Taz activity. Our results in terms of variations of the reporter signal, after both genetic and pharmacological modulation of the Wnt pathway activity, are in accordance with the model recently depicted in vitro.
The general and sustained reporter activity we observed in the endothelium during embryogenesis suggested a functional involvement of Yap/Taz signaling in developmental angiogenesis. Yap/Taz knockdown impaired the intersegmental vessels (ISVs) growth, while the overactivation of Yap/Taz-mediated transcription caused an aberrant sprouting from the ISVs. The vessel sprouting-promoting capacity of Yap/Taz is cell-autonomous, as the same phenomenon was observed by expressing TAZ-4SA under the control of an endothelium-specific promoter.
The CTGF-based zebrafish reporter is a new powerful tool to study in vivo Yap/Taz pathway activation, with possible applications in drug screening, regeneration and cancer biology. It permitted to confirm in vivo during development the crosstalk between Wnt/β-catenin and Yap/Taz pathways and to discover a novel role of Yap/Taz in vessel sprouting, suggesting a pro-angiogenic function of YAP/TAZ transcriptional activity
Time to fight: targeting the circadian clock molecular machinery in cancer therapy
No full textThe circadian clock regulates a wide range of molecular pathways and biological processes. The expression of clock genes is often altered in cancer, fostering tumor initiation and progression. Inhibition and activation of core circadian clock genes, as well as treatments that restore circadian rhythmicity, have been successful in counteracting tumor growth in different experimental models. Here, we provide an up-to-date overview of studies that show the therapeutic effects of targeting the clock molecular machinery in cancer, both genetically and pharmacologically. We also highlight future areas for progress that offer a promising path towards innovative anticancer strategies. Substantial limitations in the current understanding of the complex interplay between the circadian clock and cancer in vivo need to be addressed in order to allow clock-targeting therapies in cancer
TUTELA DEL LAVORO E LIBERTA' D'IMPRESA NEI PROCESSI DI ESTERNALIZZAZIONE
No full textL’elaborato analizza le conseguenze lavoristiche della successione fra imprenditori, muovendo da una ricognizione delle varie tipologie di esternalizzazione con le relative esigenze e principali criticità.
L’indagine si concentra in primo luogo sul trasferimento d’azienda, esaminando la normativa e la giurisprudenza europee per passare poi alla disciplina di diritto interno, alle procedure sindacali e a uno specifico focus sul trasferimento delle aziende in crisi.
Successivamente l’autore si sofferma sull’appalto, prendendone in particolare considerazione gli indici di genuinità, i criteri di distinzione dalla somministrazione illecita di manodopera e la tutela delle maestranze in caso di avvicendamento fra imprese.
Da ultimo, la ricerca approfondisce le c.d. “clausole sociali”, sia di prima che di seconda generazione, valutandone la compatibilità con il diritto eurounitario e con la costituzione nonché riflettendo sui possibili rimedi in caso di loro violazione.The author analyzes the labour consequences of the succession between entrepreneurs, starting from a recognition of the various types of outsourcing with the related needs and main critical issues.
The survey focuses primarily on the transfer of businesses, examining European legislation and case-law and then moving on to internal legislation, trade union procedures and a specific focus on the transfer of companies in crisis.
The author then dwells on the contract, taking into account in particular the indications of authenticity, the criteria of distinction from the illicit administration of labour and the protection of workers in the event of turnover between companies.
Finally, the research deepens the "social clauses", both first and second generation, assessing their compatibility with European law and with the constitution and reflecting on possible remedies in case of their violation
Ultra Low Carbon Vehicles: New Parameters for Automotive Design
No full textAs the influence of vehicle emissions on our environment has become better understood, the UK government has recently placed urgent emphasis on the implementation of low carbon
technologies in the automotive industry through: the UK Low Carbon Industrial Strategy. The overall objective is to offer big incentives to consumers and support for the development of infrastructure and engineering solutions. This scheme however does not consider how the development of functional and experiential user value might drive consumer demand, contributing to the adoption of low carbon vehicles (LCVs) in the mass market.
With the emergence of the North East of England as the UK’s first specialised region for the development of ultra-low carbon vehicles (ULCVs), ONE North East, as a development agency for the region's economic and business development, and Northumbria University Ideas-lab have supported a project to facilitate innovation through the collaboration of technology, research and development (R&D) and business. The High Value Low Carbon (HVLC) project aims to envisage
new user value made possible by the integration of low carbon vehicle platforms with new process and network technologies. The HVLC consortium represents vehicle manufacturers and their suppliers as well as technology based companies and through an ongoing process of design concept generation the project offers a hub for innovation led enterprise.
Whilst new technological developments in areas such as power generation, nano materials, hydrogen fuel cells, printed electronics and networked communications will all impact on future automotive design, the mass adoption of low carbon technologies represents a paradigm shift for the motorist. This paper aims to describe how the mapping of new parameters will lead to new transport scenarios that will create the space for new collaborative research on user experiences supported by innovative technologies and related services
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Matteo Maria Boiardo (1441-1494)
No full textMatteo Maria Boiardo – contemporary of Sir Thomas Malory and forerunner of Shakespeare, Spenser, Milton, Cervantes, Tolkien, and C.S. Lewis – is best known as the author of the Orlando Innamorato [Orlando in Love], a poem in ottava rima organically merging Carolingian epic and Arthurian romance. He was also a lyric and pastoral poet, playwright, and translator of classical texts into the Italian vernacular. Matteo was active in Italy during the years of 1463 throughout 1494, the year of his death
Uno sguardo all'Est. Lavoro e diritti fondamentali in Russia e negli altri Paesi della Comunità di Stati indipendenti
No full textNel contributo l'autore, dopo aver esaminato i diritti fondamentali del lavoro presenti nelle Costituzioni dei Paesi della Comunità di Stati indipendenti, si sofferma sul diritto del lavoro russo. La trattazione, a carattere diacronico, abbraccia i rami principali del diritto del lavoro: il rapporto individuale, le relazioni collettive, il mercato del lavoro. L'autore rileva una sorprendente continuità nelle linee essenziali del diritto del lavoro russo, pur nel susseguirsi di sistemi socioeconomici completamente diversi. Egli conclude che il diritto del lavoro russo appartiene saldamente alla matrice europea della materia.The contribution of the author starts by examining fundamental labour rights recognised by the Constitutions of the Countries of the Community of Independent States. Then the author focuses upon Russian Labour Law, going through the history of the main branches of the discipline: the individual relationship, industrial relations, the labour market. The article finds a surprising continuity in the essential lines of Russian Labour Law, in spite of the completely different socio-economic systems that were experimented in Russia during the twentieth century. The author concludes that Russian Labour Law clearly belongs to the European matrix of the discipline
Corpo, spazio, architettura. Fenomenologia dell’esperienza spaziale Matteo Vegetti, Fabrizia Bandi (eds.), Editrice Morcelliana, Brescia 2024
No full textThe author reviews "Corpo, spazio, architettura: Fenomenologia dell’esperienza spaziale," edited by Matteo Vegetti and Fabrizia Bandi. The book examines the philosophical underpinnings of spatial experience through a phenomenological lens, engaging with both traditional and contemporary notions of how space, body, and architecture interconnect. The author highlights the text\u27s exploration of how architects and designers might integrate phenomenological insights into their work, transforming space from mere physical dimensions into lived, experienced reality. The review appreciates the book’s thematic organization, which starts with foundational theories from Merleau-Ponty and expands through discussions on virtual spaces and the future of architecture in the digital age. Through this anthology, readers gain a comprehensive view of the architectural implications of phenomenological thought, making it a critical resource for anyone involved in the conceptualization and design of space
The circadian protein BMAL1 supports endothelial cell cycle during angiogenesis
No full textAims The circadian clock is an internal biological timer that co-ordinates physiology and gene expression with the 24-h solar day. Circadian clock perturbations have been associated to vascular dysfunctions in mammals, and a function of the circadian clock in angiogenesis has been suggested. However, the functional role of the circadian clock in endothelial cells (ECs) and in the regulation of angiogenesis is widely unexplored. Methods and results Here, we used both in vivo and in vitro approaches to demonstrate that ECs possess an endogenous molecular clock and show robust circadian oscillations of core clock genes. By impairing the EC-specific function of the circadian clock transcriptional activator basic helix-loop-helix ARNT like 1 (BMAL1) in vivo, we detect angiogenesis defects in mouse neonatal vascular tissues, as well as in adult tumour angiogenic settings. We then investigate the function of circadian clock machinery in cultured EC and show evidence that BMAL and circadian locomotor output cycles protein kaput knock-down impair EC cell cycle progression. By using an RNA- and chromatin immunoprecipitation sequencing genome-wide approaches, we identified that BMAL1 binds the promoters of CCNA1 and CDK1 genes and controls their expression in ECs. Conclusion(s) Our findings show that EC display a robust circadian clock and that BMAL1 regulates EC physiology in both developmental and pathological contexts. Genetic alteration of BMAL1 can affect angiogenesis in vivo and in vitro settings
Wnt/β-Catenin Signaling Regulates Yap/Taz Activity during Embryonic Development in Zebrafish
No full text: Hippo-YAP/TAZ and Wnt/β-catenin signaling pathways, by controlling proliferation, migration, cell fate, stemness, and apoptosis, are crucial regulators of development and tissue homeostasis. We employed zebrafish embryos as a model system to elucidate in living reporter organisms the crosstalk between the two signaling pathways. Co-expression analysis between the Wnt/β-catenin Tg(7xTCF-Xla.Siam:GFP)ia4 and the Hippo-Yap/Taz Tg(Hsa.CTGF:nlsmCherry)ia49 zebrafish reporter lines revealed shared spatiotemporal expression profiles. These patterns were particularly evident in key developmental regions such as the midbrain-hindbrain boundary (MHB), epidermis, muscles, neural tube, notochord, floorplate, and otic vesicle. To investigate the relationship between the Wnt/β-catenin pathway and Hippo-Yap/Taz signaling in vivo, we conducted a series of experiments employing both pharmacological and genetic strategies. Modulation of the Wnt/β-catenin pathway with IWR-1, XAV939, or BIO resulted in a significant regulation of the Yap/Taz reporter signal, highlighting a clear correlation between β-catenin and Yap/Taz activities. Furthermore, genetic perturbation of the Wnt/β-catenin pathway, by APC inhibition or DKK1 upregulation, elicited evident and robust alteration of Yap/Taz activity. These findings revealed the intricate regulatory mechanisms underlying the crosstalk between the Wnt/β-catenin and Hippo-Yap/Taz signaling, shedding light on their roles in orchestrating developmental processes in vivo
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