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The MET oncogene encodes a tyrosine kinase (TK) receptor. Its activation protects cells from death but also stimulates DNA damage response by triggering excess replicative stress. Transcriptomic classification of cancer cell lines based on MET expression showed that response to the PARP inhibitor (PARPi) olaparib is poorer in MET overexpressing cell lines. Accordingly, a high MET expressing lung carcinoma cell line was sensitized to PARPi by MET TK inhibition. This was not linked solely to MET overexpression: other MET overexpressing cell lines were biochemically but not functionally responsive to combined inhibition. Moreover, exogenously induced MET overexpression was unable to induce resistance to PARPi. The MET overexpressing cell line, responsive to the combined PARP and MET inhibition, carried a heterozygous mutation of the ATM gene and showed an attenuated response of ATM to PARPi. Among the downstream targets of ATM activation, NuMA was phosphorylated only in response to the combined PARP and MET inhibition. Given the role played by NuMA in mitosis, data show that the latter is affected by MET and PARP inhibition in cells with haploinsufficient ATM. This is important as ATM heterozygous mutation is frequently found in human cancer and in lung carcinomas in particular.

Oncostatin M (OSM) is a pleiotropic cytokine of the interleukin (IL)-6 family that contributes to the progression of chronic liver disease. Here we investigated the role of OSM in the development and progression of hepatocellular carcinoma (HCC) in non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH). The role of OSM was investigated in (1) selected cohorts of NAFLD/NASH HCC patients, (2) liver cancer cells exposed to human recombinant OSM or stably transfected to overexpress human OSM, (3) murine HCC xenografts, and (4) a murine NASH-related model of hepatic carcinogenesis. OSM was found to be selectively overexpressed in HCC cells of NAFLD/NASH patients, depending on tumor grade. OSM serum levels, barely detectable in patients with simple steatosis or NASH, were increased in patients with cirrhosis and more evident in those carrying HCC. In this latter group, OSM serum levels were significantly higher in the subjects with intermediate/advanced HCCs and correlated with poor survival. Cell culture experiments indicated that OSM upregulation in hepatic cancer cells contributes to HCC progression by inducing epithelial-to-mesenchymal transition and increased invasiveness of cancer cells as well as by inducing angiogenesis, which is of critical relevance. In murine xenografts, OSM overexpression was associated with slower tumor growth but an increased rate of lung metastases. Overexpression of OSM and its positive correlation with the angiogenic switch were also confirmed in a murine model of NAFLD/NASH-related hepatocarcinogenesis. Consistent with this, analysis of liver specimens from human NASH-related HCCs with vascular invasion showed that OSM was expressed by liver cancer cells invading hepatic vessels. In conclusion, OSM upregulation appears to be a specific feature of HCC arising on a NAFLD/NASH background, and it correlates with clinical parameters and disease outcome. Our data highlight a novel pro-carcinogenic contribution for OSM in NAFLD/NASH, suggesting a role of this factor as a prognostic marker and a putative potential target for therapy. © 2022 The Authors. The Journal of Pathology published by John WileySons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

We report on the detection of an angiogenic molecule Vascular Endothelial Growth Factor (VEGF) in different biological matrices by means of a new integrated biosensing platform exploiting the properties of Bloch surface waves. The new platform takes advantage of a tandem configuration, in which both label-free and enhanced fluorescence detection are implemented. Specifically designed one dimensional photonic crystals were deposited directly on disposable and low cost plastic biochips. A direct sandwich immunoassay was used to detect VEGF in buffer, cell culture supernatant and human plasma at low concentration (ng/mL). The platform enabled the detection of VEGF in all three matrices with high resolution, fast turnaround time (30 min) and in close agreement with the results of enzyme linked immunosorbent assays.

Background Guidelines recommend low-dose, short-term glucocorticoid (GC) treatment for rheumatoid arthritis (RA).1,2 Long-term use, especially at prednisone-equivalent doses >5 mg/d, should be avoided,2 but many patients (pts) with early established RA receive long-term GCs, often ≥5 mg/d.3 Objectives Compare GC tapering vs continuation to maintain disease control in RA pts with chronic GC exposure receiving tocilizumab (TCZ) in a randomized controlled trial. Methods Before randomization pts had to receive TCZ ± conventional synthetic (cs)DMARDs and GC (prednisone-equivalent dose 5-15 mg/d) for ≥24 wks. At randomization pts had to be in at least low disease activity (LDA: DAS28-ESR ≤3.2) and receiving stable concomitant therapy (prednisone 5 mg/d [GC5mg]) and TCZ ± csDMARDs) for ≥4 wks. Pts were randomized to continue blinded GC5mg for 24 wks or undergo blinded taper (GCtaper, from 4 mg/d with 1-mg reduction every 4 wks to 0 mg/d at wks 16-24) while receiving stable TCZ and csDMARD doses. Pts with RA flare (DAS28-ESR >3.2 and increase >0.6 vs baseline) received open-label rescue GC5mg for 2 wks and continued blinded treatment. Primary outcome was mean change in DAS28-ESR at wk 24. Key secondary outcome was treatment success—DAS28-ESR ≤3.2 at wk 24 and no RA flare during 24 wks and no adrenal insufficiency necessitating replacement therapy. Results 259 pts were randomized to GCtaper (n=131) or GC5mg (n=128); 114 and 112, respectively, completed 24 wks. Mean baseline DAS28-ESR was 1.9. Mean RA duration was 9.2 y. For the primary endpoint, the between-arm difference was 0.6 DAS28-ESR units (95% CI, 0.3, 0.9; p Conclusion Continued GC5mg provided better DAS28-ESR control than GC taper in RA pts in LDA or remission. The 0.6 DAS28-ESR unit between-arm difference should be interpreted in the context of approximately two-thirds of tapered pts experiencing treatment success and no tapered pts discontinuing due to lack of flare control. The taper schedule was safe regarding adrenal insufficiency. The results suggest that all pts achieving LDA or remission with TCZ and receiving long-term low-dose GC should be considered for GC tapering, ideally targeting discontinuation. References [1] Singh JA et al. Arthritis Rheum. 2016;68:1. [2] Smolen JS et al. Ann Rheum Dis. 2017;76:960. [3] Buttgereit F, Bijlsma JW. Ann Rheum Dis. 2017;76:1785. Disclosure of Interests Gerd Rudiger Burmester Consultant for: Roche, Sanofi-Genzyme, Speakers bureau: Roche, Sanofi-Genzyme, Frank Buttgereit: None declared, Corrado Bernasconi Consultant for: Roche, Jose-Maria Alvaro-Gracia Consultant for: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Speakers bureau: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Nidia Castro Shareholder of: F. Hoffmann-La Roche, Employee of: F. Hoffmann-La Roche, maxime dougados Grant/research support from: Eli Lilly and Company, Pfizer, AbbVie, and UCB Pharma, Consultant for: Eli Lilly and Company, Pfizer, AbbVie, and UCB Pharma, Cem Gabay Grant/research support from: Roche, Pfizer, AB2 Bio Ltd, Consultant for: Roche, Pfizer, Lilly, AbbVie, Sanofi, Regeneron, Bristol-Myers Squibb, Novartis, UCB, AB2 Bio Ltd, Debiopharm, Jacob M. van Laar Grant/research support from: Genentech, Consultant for: F. Hoffmann-La Roche, J. Michael Nebesky Shareholder of: F. Hoffmann-La Roche, Employee of: F. Hoffmann-La Roche, Attila Pethoe-Schramm Shareholder of: F. Hoffmann-La Roche, Employee of: F. Hoffmann-La Roche, Carlo Salvarani: None declared, Marc Y. Donath Consultant for: Roche, Markus R. John Shareholder of: Roche, Employee of: Roche

Background Relapsing polychondritis (RP) is a systemic autoimmune disease characterized by inflammation of the cartilaginous tissues associated with the appearance of symptoms caused by the involvement of other non-cartilaginous tissues and the presence of systemic symptoms. Objectives To describe the clinical characteristics and treatment of patients diagnosed with RP in a third level university hospital. Methods Data from RP patients were obtained from the computerized medical history of our center, including demographics, disease phenotype, treatment, laboratory and imaging studies. Results A total of 12 patients were included; 66.7% were women, age at diagnosis was 36 [34-52.5] years (median [p25-p75]). The main manifestation observed was auricular involvement (75%) followed by nasal (66.7%), joint (66.7%), and laryngo-tracheo-bronchial disease (50%). All patients were on DMARDs treatment and two thirds of the patients were receiving biological therapy at the time of analysis. Infliximab was the first biological line in all the cases (100%). Other biological therapies used, in order of frequency, were: adalimumab (25%), rituximab (25%) and tocilizumab (12.5%). 50% of the patients associated another autoimmune disease diagnosed before or during RP follow-up: Crohn9s disease, epidermolysis bullosa, primary biliary cholangitis, autoimmune hypothyroidism, systemic lupus erythematosus, granulomatosis with polyangiitis and ulcerative colitis. Only one patient (8.33%) associated hematologic disease (myelodysplastic syndrome) and another one died during follow-up. Conclusion RP is a rare disease with a broad spectrum of clinical expression, which often requires intensive management to control its manifestations, potentially serious when there is involvement of the tracheobronchial tree or is associated with other autoimmune diseases. A large part of our patients require biological therapy at some point in their evolution. References Disclosure of Interests Pablo Moreno: None declared, DOLORES MARTINEZ-QUINTANILLA JIMENEZ: None declared, Noelia Garca Castaeda: None declared, Jose-Maria Alvaro-Gracia Consultant for: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Speakers bureau: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Isidoro Gonzlez-lvaro: None declared, Alicia Humbra: None declared, Irene Llorente : None declared, Ana Ortiz: None declared, Esther Patio: None declared, Eva Tomero Muriel: None declared, Esther Vicente: None declared, Rosario Garcia de Vicuna Grant/research support from: Abbvie, BMS, Lilly, MSD, Novartis, Roche, Consultant for: Biogen, BMS, Mylan, Pfizer, Sanofi and Sandoz, Speakers bureau: BMS, Pfizer, Lilly, Sandoz, Santos Castaeda Consultant for: Amgen, BMS, Pfizer, Lilly, MSD, Roche, Sanofi, UCB

Background Despite advances in the treatment armamentarium for rheumatoid arthritis (RA) over the last decade, several unmet needs remain within RA management. Objectives To enhance scientific knowledge and improve patient care by providing practical tools and evidence-based awareness – in line with EULAR recommendations – on how to implement optimal RA management and address identified needs. Methods The eRA (evolving the management of RA) programme convened a Steering Committee (SC) of 17 European and International experts in RA, including representation from rheumatologists, nurses, health professionals and patients. Through literature search and assessment of expert opinion, 4 core areas of unmet need were identified: delays in referral and initiation of disease-modifying anti-rheumatic drugs, inconsistent application of treat-to-target strategy, sub-optimal identification and management of comorbidities, and a growing need to actively engage patients with their care. Several clinical and educational gaps associated with these unmet needs were also identified. To fill these gaps and support application of an optimal approach to RA management, practical and educational tools were developed. A cascade process engaging national leaders in rheumatology was then initiated to disseminate the tools to health professionals across Europe, with tools being updated in participating countries to ensure alignment with local practices and individual country needs. Broader programme dissemination is supported by congress activities and a digital communication plan. The eRA programme is funded by Sanofi Genzyme. Programme direction and content creation are driven by the SC. Results A suite of 12 educational tools was developed (Figure 1), including educational slides to raise awareness of unmet needs, self-reflection questionnaires to evaluate personal practice alongside EULAR recommendations, case scenarios outlining the optimal approach to management, and clinical checklists to support daily practice. The process to disseminate these tools commenced in November 2017 with a Multi-Country Workshop, which convened 43 national leaders in rheumatology from countries participating in the programme. Feedback on the eRA tools at this meeting was positive, and in 2018, additional countries were onboarded to the programme. Launch of a digital web-platform aims to further support dissemination of the eRA tools. Conclusion The content of the eRA programme has been well-received, largely due to the programme’s flexible approach, which is sympathetic to individual country needs. As the cascade process continues, it is anticipated that many more health professionals and patients will benefit from the eRA tools. References: Disclosure of Interests: Gerd Rudiger Burmester Consultant for: Roche, Sanofi-Genzyme, Speakers bureau: Roche, Sanofi-Genzyme, Mart van de Laar Grant/research support from: AbbVie, Eli Lilly, Janssen-Cilag, Merck Inc, Pfizer Inc, Sanofi Genzyme, Speakers bureau: Eli Lilly, Pfizer Inc, Jose-Maria Alvaro-Gracia Consultant for: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Speakers bureau: AbbVie, Bristol-Myers Squibb, Eli Lilly, MSD, Novartis, Pfizer Inc, Roche, Sanofi, and UCB, Neil Betteridge Consultant for: Amgen, Eli Lilly, Grunenthal, GSK, Heart Valve Voice, Janssen, Roche, Sanofi Genzyme and Sanofi Regeneron, Speakers bureau: Amgen, Eli Lilly, Grunenthal, GSK, Heart Valve Voice, Janssen, Roche, Sanofi Genzyme and Sanofi Regeneron, Jaime Calvo Consultant for: Bristol-Myers Squibb, Janssen, Celgene, Sanofi Genzyme, Speakers bureau: Bristol-Myers Squibb, Bernard Combe Consultant for: Abbvie, Bristol-Myers Squibb, Gilead, Janssen, Eli Lilly, MSD, Novartis, Pfizer, Roche-Chugai, Sanofi, UCB, Patrick Durez Speakers bureau: Bristol-Myers Squibb, Eli Lilly, Sanofi, Celltrion, Ricardo J.O Ferreira Consultant for: Sanofi Genzyme, Bruno Fautrel Grant/research support from: AbbVie, Lilly, MSD, Pfizer, Consultant for: AbbVie, Biogen, BMS, Celgene, Janssen, Lilly, Medac, MSD, NORDIC Pharma, Novartis, Pfizer, Roche, Sanofi-Aventis, Sanofi Genzyme, SOBI, UCB, Cem Gabay Grant/research support from: Roche, Pfizer, AB2 Bio Ltd, Consultant for: Roche, Pfizer, Lilly, AbbVie, Sanofi, Regeneron, Bristol-Myers Squibb, Novartis, UCB, AB2 Bio Ltd, Debiopharm, Annamaria Iagnocco: None declared, Carlomaurizio Montecucco Speakers bureau: AbbVie, Bristol-Myers Squibb, Celgene, Sanofi, Genzyme, Lilly, MSD, Pfizer, UCB, Mikkel Ostergaard Grant/research support from: Abbvie, Celgene, Centocor, Merck, Novartis, Consultant for: Abbvie, BMS, Boehringer-Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo, Orion, Pfizer, Regeneron, Roche, and UCB, Speakers bureau: Abbvie, BMS, Boehringer-Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo, Orion, Pfizer, Regeneron, Roche, and UCB, Sofia Ramiro Grant/research support from: MSD, Consultant for: AbbVie, Lilly, MSD, Novartis, Pfizer, Sanofi, Speakers bureau: AbbVie, Lilly, MSD, Novartis, Pfizer, Sanofi, Andrea Rubbert-Roth Consultant for: Chugai, Eli Lilly, Roche, and Sanofi, Speakers bureau: AbbVie, Bristol-Myers Squibb, Chugai, Hexal/Novartis, Janssen, Eli Lilly, Merck Sharp & Dohme, Pfizer, Roche, and Sanofi, Tanja Stamm Grant/research support from: TS has received grant support from AbbVie., Paid instructor for: TS has received speaker fees from AbbVie, Janssen, MSD, Novartis, and Roche., Zoltan Szekanecz Grant/research support from: Pfizer, UCB, Consultant for: Pfizer, Abbvie, Roche, Sanofi, Lilly, Novartis, Speakers bureau: Pfizer, Abbvie, Roche, Sanofi, Lilly, Novartis, Peter C. Taylor Grant/research support from: Celgene, Galapagos, Eli Lilly, UCB, Consultant for: AbbVie, Galapagos, Gilead, Eli Lilly, Pfizer Inc

Background:The eRA (evolving the management of RA) programme was initiated in Europe to provide practical educational tools that address unmet needs in the management of rheumatoid arthritis (RA). Several eRA tools – covering early access to care, management of comorbidities, treat-to-target strategies, and patient empowerment – are available to the rheumatology community. Through ongoing activities, the eRA Steering Committee (SC) identified a need for tools on non-pharmacological management of RA.Objectives:To improve accessibility to eRA tools for rheumatology professionals; to review the evidence base of non-pharmacological interventions to create new eRA resources that may support management decisions.Methods:A web platform providing information on eRA programme and tools was developed in 2019. The platform collects survey-based metrics to quantify perception of eRA and use of eRA tools in clinical practice. Platform and tools are translated to further support access and use across Europe.To address unmet needs in non-pharmacological patient management, the eRA SC reviewed the core literature on agreed priority interventions, including physical activity, diet, patient education and self-management, psychosocial interventions, occupational therapy and orthotics, hand exercises, and hydrotherapy/balneotherapy. Available evidence for each intervention was assessed and graded according to the Oxford Centre for Evidence-based Medicine Levels of Evidence.Results:The eRA web platform is now live in 3 countries (www.evolvingthemanagementofRA.com), hosting translated copies of the eRA tools, with additional countries launching throughout 2020.From a review of core literature on non-pharmacological interventions, the eRA SC determined that strong evidence exists to support use of physical activity, patient education and self-management, psychosocial interventions, and occupational therapy and orthotics. Evidence was lacking or conflicting for diet and nutrition, hand exercises, and balneotherapy/hydrotherapy. A set of educational slides was produced by the eRA SC to summarise the evidence (Fig. 1) and provide top-line guidance on use of interventions in practice that should engage relevant members of the multi-disciplinary team. These slides are available through eRA dissemination activities.Conclusion:The eRA programme content is now freely available to health care professionals in several countries on a web platform, supported by translations of the eRA tools. An additional slide set on non-pharmacological management serves to further increase the practical guidance of this programme’s educational offering.Acknowledgments:The eRA programme is funded by Sanofi Genzyme. Programme direction and content creation are driven by an independent Steering CommitteeDisclosure of Interests:Gerd Rüdiger Burmester Consultant of: AbbVie Inc, Eli Lilly, Gilead, Janssen, Merck, Roche, Pfizer, and UCB Pharma, Speakers bureau: AbbVie Inc, Eli Lilly, Gilead, Janssen, Merck, Roche, Pfizer, and UCB Pharma, Jose-Maria Alvaro-Gracia Grant/research support from: Abbvie, Elli-Lilly, MSD, Novartis, Pfizer, Consultant of: Abbvie, BMS, Janssen-Cilag, Elli-Lilly, MSD, Novartis, Pfizer, Sanofi, Tigenix, Roche, UCB, Paid instructor for: Elli-Lilly, Pfizer, Roche, Speakers bureau: Abbvie, BMS, Janssen-Cilag, Elli-Lilly, Gedeon Richter, MSD, Novartis, Pfizer, Sanofi, Tigenix, Roche, UCB, Neil Betteridge Consultant of: Amgen, Eli Lilly and Company, Grunenthal, GSK, Sanofi Genzyme, Jaime Calvo Grant/research support from: Lilly, UCB, Consultant of: Abbvie, Jansen, Celgene, Bernard Combe Grant/research support from: Novartis, Pfizer, Roche-Chugai, Consultant of: AbbVie; Gilead Sciences, Inc.; Janssen; Eli Lilly and Company; Pfizer; Roche-Chugai; Sanofi, Speakers bureau: Bristol-Myers Squibb; Gilead Sciences, Inc.; Eli Lilly and Company; Merck Sharp & Dohme; Pfizer; Roche-Chugai; UCB, Patrick Durez Speakers bureau: AbbVie, Bristol-Myers Squibb, Celltrion, Eli Lilly, Pfizer, Sanofi, Ricardo J. O. Ferreira Grant/research support from: Abbvie, Consultant of: Sanofi Genzyme, Amgen, MSD, Paid instructor for: UCB, Bruno Fautrel Grant/research support from: AbbVie, Lilly, MSD, Pfizer, Consultant of: AbbVie, Biogen, BMS, Boehringer Ingelheim, Celgene, Lilly, Janssen, Medac MSD France, Nordic Pharma, Novartis, Pfizer, Roche, Sanofi Aventis, SOBI and UCB, Annamaria Iagnocco Grant/research support from: Abbvie, MSD and Alfasigma, Consultant of: AbbVie, Abiogen, Alfasigma, Biogen, BMS, Celgene, Eli-Lilly, Janssen, MSD, Novartis, Sanofi and Sanofi Genzyme, Speakers bureau: AbbVie, Alfasigma, BMS, Eli-Lilly, Janssen, MSD, Novartis, Sanofi, Carlomaurizio Montecucco: None declared, Mikkel Ǿstergaard Grant/research support from: AbbVie, Bristol-Myers Squibb, Celgene, Merck, and Novartis, Consultant of: AbbVie, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo Nordisk, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, and UCB, Speakers bureau: AbbVie, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo Nordisk, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, and UCB, Sofia Ramiro Grant/research support from: MSD, Consultant of: Abbvie, Lilly, Novartis, Sanofi Genzyme, Speakers bureau: Lilly, MSD, Novartis, Andrea Rubbert-Roth Consultant of: Abbvie, BMS, Chugai, Pfizer, Roche, Janssen, Lilly, Sanofi, Amgen, Novartis, Tanja Stamm Grant/research support from: AbbVie, Roche, Consultant of: AbbVie, Sanofi Genzyme, Speakers bureau: AbbVie, Roche, Sanofi, Zoltán Szekanecz Grant/research support from: Pfizer, UCB, Consultant of: Sanofi, MSD, Abbvie, Pfizer, Roche, Novertis, Lilly, Gedeon Richter, Amgen, Peter C. Taylor Grant/research support from: Celgene, Eli Lilly and Company, Galapagos, and Gilead, Consultant of: AbbVie, Biogen, Eli Lilly and Company, Fresenius, Galapagos, Gilead, GlaxoSmithKline, Janssen, Nordic Pharma, Pfizer Roche, and UCB, Mart van de Laar Consultant of: Sanofi Genzyme, Speakers bureau: Sanofi Genzyme

The vascular endothelial growth factor-A (VEGF) plays a crucial role in tumor angiogenesis. Through its primary receptor VEGFR-2, VEGF exerts the activity of a multitasking cytokine, which is able to stimulate endothelial cell survival, invasion and migration into surrounding tissues, proliferation, as well as vascular permeability and inflammation. The core components of VEGF signaling delineate well-defined intracellular routes. However, the whole scenario is complicated by the fact that cascades of signals converge and branch at many points in VEGF signaling, thus depicting a complex signal transduction network that is also finely regulated by different mechanisms. In this chapter, we present a careful collection of the best-characterized VEGF-induced signal transduction pathways, attempting to offer an overview of the complexity of VEGF signaling in the context of tumor angiogenesis.

A one-dimensional photonic crystal (1DPC) based on a planar stack of dielectric layers is used as an optical transducer for biosensing, upon the coupling of TE-polarized Bloch Surface Waves (BSW). The structure is tailored with a polymeric layer providing a chemical functionality facilitating the covalent binding of orienting proteins needed for a subsequent grafting of antibodies in an immunoassay detection scheme. The polymeric layer is impregnated with Cy3 dye, in such a way that the photonic structure can exhibit an emissive behavior. The BSW-coupled fluorescence shift is used as a means for detecting refractive index variations occurring at the 1DPC surface, according to a label-free concept. The proposed working principle is successfully demonstrated in real-time tracking of protein G covalent binding on the 1DPC surface within a fluidic cell.

Dielectric multilayered structures are introduced here as building blocks of disposable chips for sensing applications. Some experimental configurations are illustrated wherein flat or patterned multilayers are employed for sensing in either label-free, spectroscopic or fluorescence operational modes. In particular, the possibility to enhance and control the emission from fluorescence markers on the chip surface is demonstrated to improve the detection limit in bio-assays for cancer diagnostics.

Quantitative detection of angiogenic biomarkers provides a powerful tool to diagnose cancers in early stages and to follow its progression during therapy. Conventional tests require trained personnel, dedicated laboratory equipment and are generally time-consuming. Herein, we propose our developed biosensing platform as a useful tool for a rapid determination of Angiopoietin-2 biomarker directly from patient plasma within 30 minutes, without any sample preparation or dilution. Bloch surface waves supported by one dimensional photonic crystal are exploited to enhance and redirect the fluorescence arising from a sandwich immunoassay that involves Angiopoietin-2. The sensing units consist of disposable and low-cost plastic biochips coated with the photonic crystal. The biosensing platform is demonstrated to detect Angiopoietin-2 in plasma samples at the clinically relevant concentration of 6 ng/mL, with an estimated limit of detection of approximately 1 ng/mL. This is the first Bloch surface wave based assay capable of detecting relevant concentrations of an angiogenic factor in plasma samples. The results obtained by the developed biosensing platform are in close agreement with enzyme-linked immunosorbent assays, demonstrating a good accuracy, and their repeatability showed acceptable relative variations.

The need for decentralized clinical tests together with the concept of time and cost saving are pushing the development of portable, miniaturized, compact biosensors with diagnostic and prognostic purpose. Here, we propose an innovative detection system based on a Single Photon Avalanche Diode (SPAD) with high sensitivity and low noise, crucial features for an efficient chemiluminescence biosensor. The SPAD detector, having 60. ?m diameter, has a Photon Detection Efficiency higher than 55% at 425. nm and a Dark Count Rate lower than 100. Hz at room temperature. Our design allows a good optical coupling efficiency between sample and detector. A specific biofunctional surface was implemented taking advantage of aptamers, short DNA sequences having high selectivity and affinity toward their targets. We successfully detected physiological levels of Vascular Endothelial Growth Factor (VEGF), a circulating protein biomarker highly correlated with cancer. The SPAD aptasensor showed a Limit of Detection (LoD) in the pM range, stability (up to 42 days) and re-usability (up to seven cycles). This compact biosensor is therefore a promising step toward the actual use of portable microdevices in diagnostics.

Innate immunity may activate paracrine circuits able to entail vascular system in the onset and progression of several chronic degenerative diseases. In particular, interleukin (IL)-12 triggers a genetic program in lymphomononuclear cells characterized by the production of interferon-γ and specific chemokines resulting in an angiostatic activity. The aim of this study is to identify molecules involved in the regulation of cell cycle in endothelial cells co-cultured with IL-12-stimulated lymphomonuclear cells. By using a transwell mediated co-culture system we demonstrated that IL-12-stimulated lymphomonuclear cells induce an arrest of endothelial cells cycle in G1, which is mainly mediated by the up-regulation of p21(Cip1/Waf1), an inhibitor of cyclin kinases. This effect requires the activation of STAT1, PKCδ and p38 MAPK, while p53 is ineffective. In accordance, siRNA-dependent silencing of these molecules in endothelial cells inhibited the increase of p21(Cip1/Waf1) and the modification in cell cycle promoted by IL-12-stimulated lymphomonuclear cells. These results indicate that the angiostatic action of IL-12-stimulated lymphomononuclear cells may lie in the capability to arrest endothelial cells in G1 phase through a mechanisms mainly based on the specific up-regulation of p21(Cip1/Waf1) induced by the combined activity of STAT1, PKCδ and p38 MAPK.

MEK and ERK are central components of the mitogen-activated protein kinase pathway. However, an accurate interaction has never been studied and accurate binding constants of the binary interaction have never been directly measured. In the present work, we studied the interaction between MEK and ERK by stopped-flow fluorescence intensity and evaluated the association and dissociation rate constants (kon and koff) from the kinetic study. We compared the results obtained by using commercial and homemade protein productions. The dissociation binding constant (Kd) value determined for the binding of MEK to ERK is in good agreement with the values obtained from the analysis of the kinase enzymatic reaction in previous in vitro studies.