85 research outputs found
Selection of peptides with affinity for the N-terminal domain of GATA-1: identification of a potential interacting protein
METRO - The role and future perspectives of Cohesion Policy in the planning of Metropolitan Areas and Cities. Policy brief: The role of metropolitan areas in the EU cohesion policy
This policy brief focuses on the role that metropolitan area plays, and may play, in the development, management and implementation of the EU cohesion policy. The information it includes is drawn from the ESPON METRO project, and in particular from the 9 in-depth case studies that have been analysed through the project: Barcelona Metropolitan Area, Brno Metropolitan Area, Brussels-Capital Region, Metropolitan City of Florence, Metropolitan Area of Gdańsk-Gdynia-Sopot, Lisbon Metropolitan Area, Métropole de Lyon, Riga Metropolitan Area, Metropolitan City of Turin. More in detail, the provided information discusses the different levels of engagement of metropolitan areas around Europe with the different stages of the EU cohesion policy development, the various programmes and instruments that have been put in place in different contexts as well as the coordination mechanisms that, in different metropolitan areas, have been put in place to ensure a stronger coherence between metropolitan governance and policy and the EU cohesion policy. The document also reflects on the engagement of the business actors and the civil society, as well as on the role that metropolitan areas are playing in the framework of the Recovery and Resilience Facility
An Air-well sparging minifermenter system for high-throughput protein production.
BackgroundOver the last few years High-Throughput Protein Production (HTPP) has
played a crucial role for functional proteomics. High-quality, high yield and
fast recombinant protein production are critical for new HTPP technologies.
Escherichia coli is usually the expression system of choice in protein production
thanks to its fast growth, ease of handling and high yields of protein produced.
Even though shake-flask cultures are widely used, there is an increasing need for
easy to handle, lab scale, high throughput systems.ResultsIn this article we
described a novel minifermenter system suitable for HTPP. The Air-Well
minifermenter system is made by a homogeneous air sparging device that includes
an air diffusion system, and a stainless steel 96 needle plate integrated with a
96 deep well plate where cultures take place. This system provides aeration to
achieve higher optical density growth compared to classical shaking growth
without the decrease in pH value and bacterial viability. Moreover the yield of
recombinant protein is up to 3-fold higher with a considerable improvement in the
amount of full length proteins.ConclusionsHigh throughput production of hundreds
of proteins in parallel can be obtained sparging air in a continuous and
controlled manner. The system used is modular and can be easily modified and
scaled up to meet the demands for HTPP
METRO - The role and future perspectives of Cohesion Policy in the planning of Metropolitan Areas and Cities. Annex I: Conceptual framework and methodology
The scope of the ESPON METRO project is rather broad, as its research positions at the intersection of a number of fields, ranging from territorial governance and spatial planning, to public administration and policy analysis and European integration studies, up to regional development studies.
In order to explore the role that metropolitan areas play as catalysts and drivers of global development, as a consequence of complex processes of socioeconomic reorganisation and rescaling that have evolved through time, and with particular reference to the European Union (EU) cohesion policy, the METRO research team has framed its action and analysis within a composite and articulated conceptual and methodological framework. In particular, the latter has been shaped in order to allow the researchers engaged in the project to answer the three main policy questions animating the study:
PQ1 | What role do metropolitan areas play in the development, management and implementation of the cohesion policy?
PQ2 | What is the added value of the cohesion policy in the planning and implementation of metropolitan policies?
PQ3 | What role does the cohesion policy play in consolidating metropolitan governance and cooperation?
The conceptual and methodological framework for the project has been developed during the first months of the research, building on the materials already included in the project proposal, that were further detailed and consolidated through:
A thorough consideration of previous research works on similar matters as well as of the existing scientific literature in the field of metropolitan governance and European integration and Europeanisation.
The interaction with the project’s Steering Committee Members during the METRO kick-off meeting (October the 9th, 2020), the comments received in response to the Delivery n.1 and the outcomes
of the Steering Committee Meeting n. 2 (November the 16th, 2020) and 3 (February the 23rd, 2021).
This Annex to the Final Report presents said conceptual framework and methodology more in detail
Angiogenic potential in biological hydrogels
Hydrogels are three-dimensional (3D) materials able to absorb and retain water in large amounts while maintaining their structural stability. Due to their considerable biocompatibility and similarity with the body\u2019s tissues, hydrogels are one of the most promising groups of biomaterials. The main application of these hydrogels is in regenerative medicine, in which they allow the formation of an environment suitable for cell differentiation and growth. Deriving from these hydrogels, it is, therefore, possible to obtain bioactive materials that can regenerate tissues. Because vessels guarantee the right amount of oxygen and nutrients but also assure the elimination of waste products, angiogenesis is one of the processes at the base of the regeneration of a tissue. On the other hand, it is a very complex mechanism and the parameters to consider are several. Indeed, the factors and the cells involved in this process are numerous and, for this reason, it has been a challenge to recreate a biomaterial able to adequately sustain the angiogenic process. However, in this review the focal point is the application of natural hydrogels in angiogenesis enhancing and their potential to guide this process
Pseudo-forces in quantum mechanics
Dynamical evolution is described as a parallel section on an infinite
dimensional Hilbert bundle over the base manifold of all frames of reference.
The parallel section is defined by an operator-valued connection whose
components are the generators of the relativity group acting on the base
manifold. In the case of Galilean transformations we show that the property
that the curvature for the fundamental connection must be zero is just the
Heisenberg equations of motion and the canonical commutation relation in
geometric language. We then consider linear and circular accelerating frames
and show that pseudo-forces must appear naturally in the Hamiltonian.Comment: 6 pages, 1 figure, revtex, new section added, to appear in PR
METRO - The role and future perspectives of Cohesion Policy in the planning of Metropolitan Areas and Cities. Annex II: The role of Metropolitan areas within the EU cohesion policy
This Annex to the final Report presents and discusses in a comparative manner the evidence collected in the nine case studies that have been explored in the framework of the ESPON Targeted Analysis METRO – The role and future perspectives of cohesion policy in the planning of Metropolitan Areas and Cities (Annexes III to XI). More in detail, the document synthesizes and compares the information collected by the various research teams through the application of the project’s analytical protocol and as a consequence of their continuous interaction with the respective stakeholders.
The report is organised following the three main policy questions that have been driving the analysis:
PQ1 | What role do metropolitan areas and cities play in the development, management and implementation of the European Union (EU) cohesion policy?
PQ2 | What is the added value of the EU cohesion policy in the planning and implementation of metropolitan policies?
PQ3 | What role does the EU cohesion policy play in consolidating metropolitan governance and cooperation?
These questions are answered through the comparative analysis and assessment of the territorial and institutional contexts in which the nine stakeholders involved in the projects are active: Metropolitan City of Turin (CMTo), Barcelona Metropolitan Area (AMB), Lisbon Metropolitan Area (LMA), Brno Metropolitan Area (BMA), Metropolitan Area of Gdańsk-Gdynia-Sopot (MAG), Metropolitan City of Florence (CMFi), Métropole de Lyon (MdL), Brussels-Capital Region (BCR), Riga Metropolitan Area (RMA)
The RNA-binding protein ILF3 binds to transposable element sequences in SINEUP lncRNAs
Transposable elements (TEs) compose about half of the mammalian genome and, as embedded sequences, up to 40% of long noncoding RNA (lncRNA) transcripts. Embedded TEs may represent functional domains within lncRNAs, providing a structured RNA platform for protein interaction. Here we show the interactome profile of the mouse inverted short interspersed nuclear element (SINE) of subfamily B2 (invSINEB2) alone and embedded in antisense (AS) ubiquitin C-terminal hydrolase L1 (Uchl1), an lncRNA that is AS to Uchl1 gene. AS Uchl1 is the representative member of a functional class of AS lncRNAs, named SINEUPs, in which the invSINEB2 acts as effector domain (ED)-enhancing translation of sense protein-coding mRNAs. By using RNA-interacting domainome technology, we identify the IL enhancer-binding factor 3 (ILF3) as a protein partner of AS Uchl1 RNA. We determine that this interaction is mediated by the RNA-binding motif 2 of ILF3 and the invSINEB2. Furthermore, we show that ILF3 is able to bind a free right Arthrobacter luteus (Alu) monomer sequence, the embedded TE acting as ED in human SINEUPs. Bioinformatic analysis of Encyclopedia of DNA Elements-enhanced cross-linking immunoprecipitation data reveals that ILF3 binds transcribed human SINE sequences at transcriptome-wide levels. We then demonstrate that the embedded TEs modulate AS Uchl1 RNA nuclear localization to an extent moderately influenced by ILF3. This work unveils the existence of a specific interaction between embedded TEs and an RNA-binding protein, strengthening the model of TEs as functional modules in lncRNAs.-Fasolo, F., Patrucco, L., Volpe, M., Bon, C., Peano, C., Mignone, F., Carninci, P., Persichetti, F., Santoro, C., Zucchelli, S., Sblattero, D., Sanges, R., Cotella, D., Gustincich, S. The RNA-binding protein ILF3 binds to transposable element sequences in SINEUP lncRNAs
High-throughput assessment of the antibody profile in ovarian cancer ascitic fluids
The identification of effective biomarkers for early diagnosis, prognosis, and response to treatments remains a challenge in ovarian cancer (OC) research. Here, we present an unbiased high-throughput approach to profile ascitic fluid autoantibodies in order to obtain a tumor-specific antigen signature in OC. We first reported the reactivity of immunoglobulins (Igs) purified from OC patient ascites towards two different OC cell lines. Using a discovery set of Igs, we selected tumor-specific antigens from a phage display cDNA library. After biopanning, 700 proteins were expressed as fusion protein and used in protein array to enable large-scale immunoscreening with independent sets of cancer and noncancerous control. Finally, the selected antigens were validated by ELISA. The initial screening identified eight antigenic clones: CREB3, MRPL46, EXOSC10, BCOR, HMGN2, HIP1R, OLFM4, and KIAA1755. These antigens were all validated by ELISA in a study involving ascitic Igs from 153 patients (69 with OC, 34 with other cancers and 50 without cancer), with CREB3 showing the highest sensitivity (86.95%) and specificity (98%). Notably, we were able to identify an association between the tumor-associated (TA) antibody response and the response to a first-line tumor treatment (platinum-based chemotherapy). A stronger association was found by combining three antigens (BCOR, CREB3, and MRLP46) as a single antibody signature. Measurement of an ascitic fluid antibody response to multiple TA antigens may aid in the identification of new prognostic signatures in OC patients and shift attention to new potentially relevant targets
SINEUPs: A novel toolbox for RNA therapeutics
RNA molecules have emerged as a new class of promising therapeutics to expand the range of druggable targets in the genome. In addition to 'canonical' protein-coding mRNAs, the emerging richness of sense and antisense long non-coding RNAs (lncRNAs) provides a new reservoir of molecular tools for RNA-based drugs. LncRNAs are composed of modular structural domains with specific activities involving the recruitment of protein cofactors or directly interacting with nucleic acids. A single therapeutic RNA transcript can then be assembled combining domains with defined secondary structures and functions, and antisense sequences specific for the RNA/DNA target of interest. As the first representative molecules of this new pharmacology, we have identified SINEUPs, a new functional class of natural antisense lncRNAs that increase the translation of partially overlapping mRNAs. Their activity is based on the combination of two domains: An embedded mouse inverted SINEB2 element that enhances mRNA translation (effector domain) and an overlapping antisense region that provides specificity for the target sense transcript (binding domain). By genetic engineering, synthetic SINEUPs can potentially target any mRNA of interest increasing translation and therefore the endogenous level of the encoded protein. In this review, we describe the state-of-the-art knowledge of SINEUPs and discuss recent publications showing their potential application in diseases where a physiological increase of endogenous protein expression can be therapeutic
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