Acid microenvironment promotes cell survival of human bone sarcoma through the activation of cIAP proteins and NF-κB pathway

Extracellular acidification is a very common cause of stress in tumor microenvironment and of Darwinian pressure. In acid areas of the tumor, most cancer cells are-albeit slowly proliferating-more resistant to cell death than those in well-perfused regions. Tumor acidosis can directly regulate the expression of pro-survival proteins since a low extracellular pH activates the caspase-dependent cell death machinery. This mechanism has never been explored in bone sarcomas. We cultured osteosarcoma and Ewing sarcoma cells under low pH (pH 6.5), and we performed deep-sequencing and protein analysis. Both in in vitro and in vivo models, acidification activity enhanced tumor cells survival. However, we did not observe any change in ERK1 phosphorylation. On the contrary, both at the mRNA and protein level, we found a significant induction of TRAF adaptor proteins and of cIAP proteins (BIRC2 and/or BIRC3). As a consequence, the downstream nuclear transcription factor kappa B (NF-κB) survival pathway was increased. Furthermore, the treatment with the cIAP inhibitor LCL161 reverted the protection from apoptosis under low pH. In vitro results were confirmed both in Ewing sarcoma xenograft and in osteosarcoma patients, since the analysis of tumor tissues demonstrated that the levels of expression of TRAF1 or NF-κB1 significantly correlate with the level of expression of the vacuolar ATPase (V-ATPase), the most important proton pump in eukaryotes. Moreover, in the tissue sections of xenograft model, the nuclear translocation of RelB, a key subunit of the NF-κB transcriptional complex, localized in the tumor region that also corresponded to the acid microenvironment associated with the highest levels of expression of LAMP2 and V-ATPase, in the internal area of the tumor, as revealed by immunohistochemistry. Our data confirm that tumor acid microenvironment activates a stress-regulated switch to promote cell survival of bone sarcoma, and support the hypothesis that this mechanism is mediated by the recruitment of TRAF/cIAP complexes. Altogether, these results suggest that TRAF/cIAP can be considered as a target for anti-cancer therapies

Engineering donor mesenchymal cells with IL-7 hastens naive T cell recruitment in vitro and supports immunologic reconstitution after HSCT in NOD/SCID mice

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117: Naive and memory T regulatory cells respond to mesenchymal cells regulation

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Materials characterisation and software tools as key enablers in Industry 5.0 and wider acceptance of new methods and products

Recently, the NMBP-35 Horizon 2020 projects -NanoMECommons, CHARISMA, and Easi-stress -organised a collaborative workshop to increase awareness of their contributions to the industry "commons" in terms of characterisation and digital transformation. They have established interoperability standards for knowledge management in characterisation and introduced new solutions for materials testing, aided by the standardisation of faster and more accurate assessment methods. The lessons learned from these projects and the discussions during the joint workshop emphasised the impact of recent developments and emerging needs in the field of characterisation. Specifically, the focus was on enhancing data quality through harmonisation and stand-ardisation, as well as making advanced technologies and instruments accessible to a broader community with the goal of fostering increased trust in new products and a more skilled society. Experts also highlighted how characterisation and the corresponding experimental data can drive future innovation agendas towards tech-nological breakthroughs. The focus of the discussion revolved around the characterisation and standardisation processes, along with the collection of modelling and characterisation tools, as well as protocols for data ex-change. The broader context of materials characterisation and modelling within the materials community was explored, drawing insights from the Materials 2030 Roadmap and the experiences gained from NMBP-35 pro-jects. This whitepaper has the objective of addressing common challenges encountered by the materials com-munity, illuminating emerging trends and evolving techniques, and presenting the industry's perspective on emerging requirements and past success stories. It accomplishes this by providing specific examples and high-lighting how these experiences can create fresh opportunities and strategies for newcomers entering the market. These advancements are anticipated to facilitate a more efficient transition from Industry 4.0 to 5.0 during the industrial revolution

Novel NPM1 exon 5 mutations and gene fusions leading to aberrant cytoplasmic nucleophosmin in AML

Nucleophosmin (NPM1) mutations in acute myeloid leukemia (AML) affect exon 12, but also sporadically affect exons 9 and 11, causing changes at the protein C-terminal end (tryptophan loss, nuclear export signal [NES] motif creation) that lead to aberrant cytoplasmic NPM1 (NPM1c+), detectable by immunohistochemistry. Combining immunohistochemistry and molecular analyses in 929 patients with AML, we found non–exon 12 NPM1 mutations in 5 (1.3%) of 387 NPM1c+ cases. Besides mutations in exons 9 (n = 1) and 11 (n = 1), novel exon 5 mutations were discovered (n = 3). Another exon 5 mutation was identified in an additional 141 patients with AML selected for wild-type NPM1 exon 12. Three NPM1 rearrangements (NPM1/RPP30, NPM1/SETBP1, NPM1/CCDC28A) were detected and characterized among 13 979 AML samples screened by cytogenetic/fluorescence in situ hybridization and RNA sequencing. Functional studies demonstrated that in AML cases, new NPM1 proteins harbored an efficient extra NES, either newly created or already present in the fusion partner, ensuring its cytoplasmic accumulation. Our findings support NPM1 cytoplasmic relocation as critical for leukemogenesis and reinforce the role of immunohistochemistry in predicting AML-associated NPM1 genetic lesions. This study highlights the need to develop new assays for molecular diagnosis and monitoring of NPM1-mutated AML

Chronic Myeloid Leukemia Patient's Voice About the Experience of Treatment-Free Remission Failure: Results From the Italian Sub-Study of ENESTPath Exploring the Emotional Experience of Patients During Different Phases of a Clinical Trial

Background: The main objective of this study is to gain further insights on how chronic myeloid leukemia (CML) patients involved in an interventional clinical trial with the purpose of reaching treatment free remission (TFR) phase, perceived and experienced TFR failure. TFR failure was defined for the individual patient as either not being eligible for drug discontinuation or as having relapse in the TFR phase with reintroduction of nilotinib treatment. Methods: Using a qualitative approach, out of 25 patients with CML who experienced TFR failure 14 were interviewed. Patients' views and experiences were explored using in-depth interviews, analyzed using the Interpretative Phenomenological Analysis (IPA). Results: The analysis of the interviews revealed that the experience of the diagnosis seems to have been lived as a traumatic break that has created a dichotomy, like an ambivalence in the ways in which CML patients perceived and experienced the whole disease journey, with contradictory feelings of both positive and negative emotions (e.g., a diagnosis of cancer, that is something distressing and of being afraid of, but also with a treatment and a life expectancies of which being grateful). This ambivalence of feelings was found to give meaning to the way in which patients cognitively and emotionally experienced the different steps of their disease history. Thus, four main issues, corresponding to different steps of the patients' journey, were identified: (1) the moment of the diagnosis, (2) the experience of the illness journey: disease and treatment, (3) the moment of "TFR failure," and (4) the impact of disease, treatment and relapse on the patient's life. Conclusion: This qualitative analysis helps in understanding patients' perspective, both in terms of getting access to the inner subjective experience of having CML and its strict relationship with the involvement in a trial or its cessation. Clinicians should consider that the way in which CML patients feel engaged in a clinical trial, create expectancies about TFR or experience the TFR failure is linked to the process of coping with the diagnosis, which is characterized by ambivalence

School principals' managerial behaviours and students' achievement: An empirical analysis of Italian middle schools

Purpose: This research investigates the impact of managerial practices implemented by Italian school principals on students' outcomes, using micro-data provided by the National Evaluation Committee for Education (INVALSI) for 2013/14 school year. Design/methodology/approach: Employing an educational production function, the authors regress a set of student and school's characteristics, enriched by information from a questionnaire filled by school principals to estimate student's score at grade 8 (last year of junior secondary school), also taking into account student's prior achievement (at grade 6 – first year of junior secondary school). Findings: Indicators about managerial practices have positive coefficients, but low statistical significance. Stronger associations between management variables and test scores are detected for low-SES schools. Originality/value: The research presented here is particularly innovative in the Italian context, where little evidence exists about the impact of managerial skills in education, though institutional reforms are leading towards a strengthening of school principal's leadership role. In this paper, the authors move a first step by describing managerial practices and their diffusion in different schools and geographical areas within the country. The authors focus the attention on the role of managerial practices (what principals do) and not on the managerial skills (what principals are able to do)

Identification of rigid body inertia properties from experimental frequency response

The estimate of mass properties from the massline characteristic of experimental FRFs has attracted the attention of several authors during the last decade. The problem is tipically non-linear and, in the technical literature, iterative as well as direct methods are proposed for its solution. In this paper a new direct procedure is developed. The non-linear problem is divided into a series of simpler linear problems and solved in subsequent steps. A careful analysis is performed on the conditioning of the systems in order to assess the minimum number of required measurements, necessary to solve the problem even in noisy environments. The proposed approach produces accurate results as it is shown for two real experimentally tested systems: a two and three-dimensional structure, respectively. Of course an accurate measurement of the masslines characteristics is required. An extended discussion on this point is presented

Quis custiodet ipsos custodes? How to detect and correct teacher cheating in Italian student data

The increasing diffusion of standardized assessments of students’ competences has been accompanied by an increasing need to make reliable data available to all stakeholders of the educational system (policy makers, teachers, researchers, families and students). In this light, we propose a multistep approach to detect and correct teacher cheating, which decreases the quality of student data offered by the Italian Institute for the Educational Evaluation of Instruction and Training. Our method integrates the “mechanistic” logic of the fuzzy clustering technique with a statistical model-based approach, and it aims to improve the detection of cheating and to correct test scores at both the class and student level. The results show a normalization of the scores and a stronger correction on data for Southern regions, where the propensity to cheat appears to be highest