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Ewing Sarcomas (EWS, OMIM#612219) presents a major challenge in pediatric oncology due to its aggressive nature and poor prognosis, particularly in metastatic cases. Genetic fusions involving the EWSR1 gene and ETS family transcription factors are common in EWS, though other rarer fusions have also been identified. Current standard techniques like immunohistochemistry have failed to fully characterize the immune tumor microenvironment of EWS, hindering insights into tumor development and treatment strategies. Recent efforts apply gene expression datasets to quantify tumor-infiltrating immune cells in EWS. Similar deconvolution techniques can be also applied to DNA methylation (DNAm) arrays, which are much more stable compared to RNA-based methods. This study aims to characterize immune cell infiltration in EWS using DNAm array data. We collected 32 EWS samples from 32 consecutive patients referred to Bambino Gesù Children’s Hospital. DNAm analysis was performed by EPIC arrays; data loading, normalization, deconvolution and survival analysis were then performed in R programming environment. We observed a higher content of dendritic cells and longer overall survival in samples with EWSR1::FLI1 translocation compared to samples with rarer fusions. Moreover, T-memory lymphocytes and monocytes emerged as a significant predictor of overall survival. This study underscores the potential of DNAm arrays in providing robust insights into EWS immune profiles, offering a promising avenue for future research. Further investigations with larger cohorts are warranted to validate these findings and explore additional immune cell types influencing EWS outcomes.

Abstract In vitro models of pediatric brain tumors (pBT) are instrumental for better understanding the mechanisms contributing to oncogenesis and testing new therapies; thus, ideally, they should recapitulate the original tumor. We applied DNA methylation (DNAm) and copy number variation (CNV) profiling to characterize 241 pBT samples, including 155 tumors and 86 pBT-derived cell cultures, considering serum vs serum-free conditions, late vs early passages, and dimensionality (2D vs 3D cultures). We performed a t-SNE classification and identified differentially methylated regions in tumors compared to cell models. Early cell cultures recapitulate the original tumor, but serum media and 2D culturing were demonstrated to significantly contribute to the divergence of DNAm profiles from the parental ones. All divergent cells clustered together acquiring a common deregulated epigenetic signature suggesting a shared selective pressure. We identified a set of hypomethylated genes shared among unfaithful cells converging on response to growth factors and migration pathways, such as signaling cascade activation, tissue organization, and cellular migration. In conclusion, DNAm and CNV are informative tools that should be used to assess the recapitulation of pBT-cells from parental tumors.

The present study describes the Italian adaptation of the Academic Stressors Scale (E-CEA) of the Academic Stress Questionnaire, evaluating the relationships with general and domain-specific well-being and verifying the significant predictors and the amount of variance explained by the “non-intellective” academic competencies on the scores of student stress dimensions. The participants are 1305 students from all the different degree courses. The Italian version of the E-CEA, composed of 38 items, showed good psychometric properties both in terms of reliability and factorial structure with good fit indices. The 6 sub-dimensions, for the most part overlapping with those of the original version of the instrument, show good construct and concurrent validity as negative relationships were found with general and domain-specific well-being indices. With regard to the regressions performed, several dimensions of “non-intellective” academic competencies turned out to be significant predictors (with negative effect) with respect to the stress levels perceived in the academic environment by university students: in particular, time organization, emotional control, the ability to relate to professors and intrinsic motivation could decrease stress levels, while dedication to study and the tendency to involve one’s parents in one’s university career seemed to increase stress levels. Regarding the practical implications of the results, suggestions are provided in supporting the career paths of students to reduce risk factors for stress development and to promote academic well-being.

Digital health, encompassing the use of digital technologies in healthcare, and telemedicine, facilitating healthcare delivery across long distances, have witnessed widespread applications across various healthcare domains. The COVID-19 pandemic accelerated the adoption of digital solutions in healthcare, overcoming barriers to access and fostering transitions to new care models. However, healthcare professionals often lack digital health competencies, necessitating targeted training initiatives. This study presents a project initiated by the Order of Physicians and Dentists of Rome, promoting a comprehensive training program in digital health for healthcare professionals. This investigation aims to describe the project, report demographic characteristics of participants, and analyze survey results on participants’ perceptions of the training program. The Erasmus+ project, titled ‘Training of Physician Trainers in Telemedicine, eHealth, and Digital Medicine,’ facilitated the digital transition of the healthcare sector through international training. The project involved structured courses, job-shadowing, and support activities in Malta and Madrid. A survey, developed using the Delphi methodology, assessed participants’ views on telemedicine. Thirty participants, selected based on merit, engaged in the project. Survey responses highlighted a strong impact on participants’ understanding of digital health concepts and increased confidence in utilizing digital tools. Notably, 85% acknowledged significant skill acquisition in healthcare digitalization. The project addressed a critical training gap among healthcare professionals, emphasizing the need for ongoing education in digital health. Despite existing recommendations, formal digital health education remains limited. The study underscores the importance of educational efforts to foster a digitalized healthcare model.

Collagen VI is an essential component of the extracellular matrix (ECM) composed by α1, α2 and α3 chains and encoded by COL6A1, COL6A2 and COL6A3 genes. Dominant negative pathogenic variants in COL6A genes result in defects in collagen VI protein and are implicated in the pathogenesis of muscular diseases, including Ullrich congenital muscular dystrophy (UCMD). Here, we designed a CRISPR genome editing strategy to tackle a dominant heterozygous deletion c.824_838del in exon 9 of the COL6A1 gene, causing a lack of secreted collagen VI in a patient’s dermal fibroblasts. The evaluation of efficiency and specificity of gene editing in treating patient’s fibroblasts revealed the 32% efficiency of editing the mutated allele but negligible editing of the wild-type allele. CRISPR-treated UCMD skin fibroblasts rescued the secretion of collagen VI in the ECM, which restored the ultrastructure of the collagen VI microfibril network. By using normal melanocytes as surrogates of muscle cells, we found that collagen VI secreted by the corrected patient’s skin fibroblasts recovered the anchorage to the cell surface, pointing to a functional improvement of the protein properties. These results support the application of the CRISPR editing approach to knock out COL6A1 mutated alleles and rescue the UCMD phenotype in patient-derived fibroblasts.

BackgroundCollagen is one of the major proteins of the skin and it is particularly important for its strength and resilience. Skin aging is a natural process that is characterized by the decrease and fragmentation of collagen in the dermis. Oral supplementation with collagen peptides has been clinically shown to have a positive effect on the skin condition. However, the mechanisms of aging-related changes synthesized by cells exposed to collagen are currently not well understood. Therefore, in this in vitro study, the mechanisms associated with collagen, elastin, and versican in human dermal fibroblasts were investigated after exposure to collagen peptides.MethodsThe effects of different concentrations of collagen peptides on cell viability and metabolism were analyzed. For gene expression analysis, human dermal fibroblasts were treated with collagen peptides. This was then followed by RNA extraction and DNA synthesis. Gene expressions of collagen type 1 (COL1A1), elastin (ELN), and versican (VCAN) were quantified by quantitative reverse transcription polymerase chain reaction (RT-qPCR). In addition, collagen levels were analyzed by confocal scanning laser microscopy using immunostaining.ResultsCollagen peptides tested in the study increased the expression of the relevant COL1A1, ELN, and VCAN genes in human dermal fibroblasts (p

Abstract Background Malignant peripheral nerve sheath tumors (MPNSTs) account for 3–10% of pediatric sarcomas, 50% of which occur in neurofibromatosis type 1 (NF1). Sporadic MPNSTs diagnosis may be challenging due to the absence of specific markers, apart from immunohistochemical H3K27me3 loss. DNA methylation (DNAm) profiling is a useful tool for brain and mesenchymal neoplasms categorization, and MPNSTs exhibit a specific DNAm signature. An MPNST-like group has recently been recognized, including pediatric tumors with retained H3K27me3 mark and clinical/histological features not yet well explored. This study aims to characterize the DNAm profile of pediatric/juvenile MPNSTs/MPNST-like entities and its diagnostic/prognostic relevance. Results We studied 42 tumors from two groups. Group 1 included 32 tumors histologically diagnosed as atypical neurofibroma (ANF) (N = 5) or MPNST (N = 27); group 2 comprised 10 tumors classified as MPNST-like according to Heidelberg sarcoma classifier. We performed further immunohistochemical and molecular tests to reach an integrated diagnosis. In group 1, DNAm profiling was inconclusive for ANF; while, it confirmed the original diagnosis in 12/27 MPNSTs, all occurring in NF1 patients. Five/27 MPNSTs were classified as MPNST-like: Integrated diagnosis confirmed MPNST identity for 3 cases; while, the immunophenotype supported the change to high-grade undifferentiated spindle cell sarcoma in 2 samples. The remaining 10/27 MPNSTs variably classified as schwannoma, osteosarcoma, BCOR-altered sarcoma, rhabdomyosarcoma (RMS)-MYOD1 mutant, RMS-like, and embryonal RMS or did not match with any defined entity. Molecular analysis and histologic review confirmed the diagnoses of BCOR, RMS-MYOD1 mutant, DICER1-syndrome and ERMS. Group 2 samples included 5 high-grade undifferentiated sarcomas/MPNSTs and 5 low-grade mesenchymal neoplasms. Two high-grade and 4 low-grade lesions harbored tyrosine kinase (TRK) gene fusions. By HDBSCAN clustering analysis of the whole cohort we identified two clusters mainly distinguished by H3K27me3 epigenetic signature. Exploring the copy number variation, high-grade tumors showed frequent chromosomal aberrations and CDKN2A/B loss significantly impacted on survival in the MPNSTs cohort. Conclusion DNAm profiling is a useful tool in diagnostic work-up of MPNSTs. Its application in a retrospective series collected during pre-molecular era contributed to classify morphologic mimics. The methylation group MPNST-like is a ‘hybrid’ category in pediatrics including high-grade and low-grade tumors mainly characterized by TRK alterations.

The problem of Power Quality analysis is becoming crucial to ensuring the proper functioning of complex systems and big plants. In this regard, it is essential to rapidly detect anomalies in voltage and current signals to ensure a prompt response and maximize the system’s availability with the minimum maintenance cost. In this paper, anomaly detection algorithms based on machine learning, such as One Class Support Vector Machine (OCSVM), Isolation Forest (IF), and Angle-Based Outlier Detection (ABOD), are used as a first tool for rapid and effective clustering of the measured voltage and current signals directly on-line on the sensing unit. If the proposed anomaly detection algorithm detects an anomaly, further investigations using suitable classification algorithms are required. The main advantage of the proposed solution is the ability to rapidly and efficiently detect different types of anomalies with low computational complexity, allowing the implementation of the algorithm directly on the sensor node used for signal acquisition. A suitable experimental platform has been established to evaluate the advantages of the proposed method. All the different models were tested using a consistent set of hyperparameters and an output dataset generated from the principal component analysis technique. The best results achieved included models reaching 100% recall and a 92% F1 score.

Currently, there are a few detailed guidelines on the overall management of children and adolescents with moderate-severe atopic dermatitis. AD ​​is a complex disease presenting with different clinical phenotypes, which require an individualized and multidisciplinary approach. Therefore, appropriate interaction between primary care pediatricians, pediatric allergists, and pediatric dermatologists is crucial to finding the best management strategy. In this manuscript, members of the Italian Society of Pediatric Allergology and Immunology (SIAIP), the Italian Society of Pediatric Dermatology (SIDerP), and the Italian Society of Pediatrics (SIP) with expertise in the management of moderate-severe atopic dermatitis have reviewed the latest scientific evidence in the field. This narrative review aims to define a pathway to appropriately managing children and adolescents with moderate-severe atopic dermatitis.

(1) Background: To improve academic performance and prevent dropouts, many studies have investigated the effects of non-intellective competencies on performance, and the effects of performance on school satisfaction. The aim of this study was to investigate the direct role of both non-intellective competencies and performance on school satisfaction at the same time. (2) Methods: The study involved 731 Italian students, attending three different high schools, who responded to the H-Comp Scale, a questionnaire assessing twelve different aspects of students' skills, attitudes and motivations over the study, and the H-Sat Scale, a questionnaire assessing five different areas of school satisfaction. (3) Results: We found a strong role of non-intellective competencies on school satisfaction and a marginal role of academic performance, gender and the attended class, which reduced as students' seniority increased. (4) Conclusions: This study showed that school satisfaction depends on students' self-perception in terms of their study competencies and the motivations possessed more than their sole performance, suggesting that this helps students to improve their non-intellective competencies in order to increase their performance and reduce the risk of dropouts, both directly and indirectly, through increasing their school satisfaction.

Abstract Background The Order of Physicians and Dentists of the Province of Rome aims at focusing on the satisfaction of healthcare personnel as an essential factor for the quality of medical care in the health sector. The aim of this study is to assess and prioritize the factors that can be linked to a higher or lower degree of job satisfaction in Primary Care Pediatricians (PCPs). Methods This study is a cross sectional survey. A questionnaire was administered to all primary care pediatricians registered to the Order, exploring in particular the work activity organization, the level of satisfaction in their professional life, and the level of perceived health. A pilot activity was conducted to validate the questionnaire. Fisher exact test and ordinal logistic regression (ologit) models were used for the univariate and multivariate analysis. Results The highest level of job dissatisfaction, in both men and women, was found to be in the practice type without any form of association; among women, it reached an even higher level for those who had their own practice at a distance of 20–40 km from their home. Women, compared to men, maintained a lower level of job satisfaction also while working in Pediatric Primary Care Units (PPCUs). In PPCUs, for the same distance, females showed a more similar pattern to males. Men working in PPCUs, regardless of distance, declared a higher degree of job satisfaction. Both men and women, working as a group pediatrician or in PPCUs, did not show a significant difference in the level of job satisfaction. Conclusions The study contributes to a deeper understanding of the factors that may influence levels of career satisfaction in female and male PCPs. Therefore, research and interventions regarding job satisfaction should foster an organizational network connection among PCPs for their job and individual well-being, from a perspective of enhancing patient care. A major effort to improve work-life balance and career satisfaction among women is important, suggesting that interventions for improving job satisfaction could benefit from a gender-specific approach.