Stefano Ratti

Adult-onset Autosomal Dominant Leukodystrophy (ADLD) is a rare fatal leukodystrophy caused by increased LMNB1 gene dosage, most commonly resulting from duplication of the LMNB1 locus. Because ADLD is a gene dosage disorder, selective reduction of pathological LMNB1 expression represents a rational therapeutic strategy. Although allele-specific RNA interference has previously been shown to lower LMNB1 levels in patient-derived fibroblasts and directly reprogrammed neurons, its therapeutic effects have not been evaluated in disease-relevant human glial cells or using functional efficacy endpoints. Here, we established human induced pluripotent stem cell-derived astrocytes from ADLD patients as a human glial model in which to validate allele-specific LMNB1 silencing across molecular, cellular, and functional readouts. ADLD astrocytes recapitulated increased LMNB1 expression and characteristic nuclear abnormalities and displayed transcriptional alterations affecting extracellular matrix organization, calcium homeostasis, metabolism and RNA processing. Functionally, these cells also exhibited functional phenotypes suitable for therapeutic evaluation: astrocyte-conditioned medium impaired the viability of both murine and human oligodendroglial cultures, while conditioned-medium and direct astrocyte-seeding paradigms revealed impaired post-lesion myelin recovery in lysolecithin-treated cerebellar organotypic slices. Allele-specific LMNB1 silencing restored physiological LMNB1 levels, corrected nuclear abnormalities, attenuated astrocyte-mediated oligodendroglial toxicity, improved post-lesion myelin recovery, and was associated with selective transcriptional programs associated with extracellular support and cholesterol metabolism. Together, these findings provide molecular, cellular, and functional validation of allele-specific LMNB1 dosage correction in patient-derived human astrocytes and offer key support for LMNB1-lowering strategies in disease-relevant human glial cells.

Background: Robotic-assisted surgery requires training that integrates technical skills, visuospatial coordination, platform management, and anatomical understanding. Although synthetic and virtual-reality simulators are widely used, high-fidelity training on bodies donated to science remains comparatively underreported. This pilot study therefore aimed to evaluate participants’ perceived value of a robotic surgery training experience with the da Vinci Xi platform in a body donation setting, and to examine whether this perceived value was associated with participants’ views on the educational role of body donation. Methods: This pilot post-course mixed-methods evaluation involved 41 residents and surgeons in general surgery and urology. The program combined dry-lab and wet-lab activities using the da Vinci Xi platform and three body donors. Participants completed a study-specific questionnaire including seven Likert-type items and two open-ended questions. Quantitative data were analyzed descriptively, through Spearman correlations and exploratory subgroup comparisons; qualitative responses underwent thematic analysis. Results: Course evaluations ratings were highly positive, with mean scores ranging from 4.59 to 4.88 on a 5-point scale. The mean Training Value Index was 4.70, and the five course evaluation items showed good internal consistency. All participants expressed high willingness to attend a similar course again, and 97.6% would recommend it to colleagues. Agreement that body donation represents a gold standard for medical and surgical education and training, and its perceived ethical importance, received high ratings from 95.1 and 97.6% of participants, respectively. Both were significantly associated with the Training Value Index. Qualitative feedback highlighted hands-on robotic practice, anatomical fidelity, operative-like workflow, and requests for longer or repeated training opportunities. Conclusion: Participants perceived robotic surgery training in a body donation setting as highly valuable, and this perception was closely linked to their broader recognition of the educational value of body donation. These preliminary findings suggest that combining technical training, anatomical complexity, team-based workflow, and ethical awareness may represent a promising complement to dry-lab and virtual-reality simulation, warranting further investigation to inform the development of more integrated robotic surgery curricula.

Cell therapies represent a promising frontier in modern medicine. Mesenchymal stromal cells (MSCs) constitute a valuable source due to their minimal ethical concerns, low immunogenicity, negligible tumorigenicity in vivo, and robust paracrine activity mediated by the secretion of anti‐inflammatory and angiogenic factors. Despite these advantages, MSCs undergo rapid replicative senescence accompanied by a progressive loss of stemness markers and functional potency. In this context, cellular rejuvenation has been emerging as a pivotal strategy to restore their original biological features. These approaches aim to modulate the hallmarks of aging; as these processes are highly interconnected, multiple and complementary targets have been proposed. This review aims to discuss and analyze the main rejuvenation strategies reported in the literature for MSCs, with particular attention to approaches that can enhance their functionality and translational potential. In this context, the review focuses on five key categories of interventions: epigenetic modifications, metabolic reprogramming, modulation of cellular senescence, telomere biology, and extracellular vesicles. These strategies represent the fundamental axes through which cellular aging can be counteracted and the efficacy of cell‐based therapies improved.

Body donation is essential for medical education and research, supporting anatomical training and clinical competence. In Italy, national data on awareness of the legislation regulating body donation and factors shaping donation‐related decisions among healthcare trainees remain limited. This study investigated knowledge of the legal framework, attitudes, and willingness for body donation among Italian medical students and residents. A two‐phase mixed‐methods study was conducted. In phase 1, qualitative interviews informed the development of a structured questionnaire. In phase 2, a national cross‐sectional survey was carried out in 2023 using a CAWI questionnaire. Items explored exposure to body‐based training, awareness of body donation and related legislation, perceived barriers and facilitators, and willingness to donate. Group comparisons and multivariable logistic regression analyses were performed. A total of 1044 participants completed the survey. Most were aware of body donation (92.8%), awareness of regulating law was limited (78.2% unaware), and public information was perceived as insufficient (69.6%). Overall, 53.8% reported willingness for body donation. Cultural and spiritual beliefs, emotional discomfort, and bureaucratic procedures emerged as major barriers. Students more often identified knowledge as a facilitator and less often viewed emotional involvement as a barrier compared with residents. In multivariable analysis, willingness to donate was negatively associated with male gender and lack of exposure or interest in body‐based training. Among Italian healthcare trainees, willingness to consider body donation is high but coexists with limited legal awareness and persistent social barriers. Educational strategies within healthcare training contexts may support informed decision‐making and greater engagement.

Obstructive Sleep Apnoea (OSA) is a sleep-related chronic disorder affecting over one billion people worldwide. Mild to moderate OSA can be treated with a Mandibular Advancement Device (MAD), but its effects on airway anatomy remain unclear. The present study aims to standardize upper airway and soft tissue CT measurements and to evaluate three-dimensional changes following progressive mandibular protrusion. Eight body donors underwent CT acquisitions at baseline and at 2 mm mandibular advancement steps up to 12 mm using a MAD-like device. Upper Airway Space (AS), Tongue (T) and Floor of the Oral Cavity (CF) measurements were obtained using a standardized protocol. Inter- and intra-observer reliability were assessed using ICC. Superior Airway Space (SPAS) increased significantly starting from 4 mm, while Inferior Airway Space at Gonion (G-IAS) changed from 6 mm. Inferior portions of T and CF increased from 2 mm, whereas differences in the superior CF occurred only at 12 mm. SPAS and G-IAS appear the most responsive airway regions, while early soft tissue changes are detected in their inferior anatomical sections. High intra- and inter-observer reliability has been observed. Although limited by the model, the proposed protocol provides a reproducible and reliable framework for analysing structural effects of mandibular advancement and may support treatment optimization.

Objective: Transfer RNA-derived fragments (tRFs) belong to an emerging class of small non-coding RNAs that dynamically respond to metabolic stressors and drive different pathological processes, yet their role in osteoarthritis (OA) remains poorly explored. We aimed to define the tRF landscape in OA and investigate the function of 3’tRF ^Asp(GTC) in chondrocyte stress adaptation and translational control. Methods: Ex vivo , cartilage specimens from OA patients (n=6) and healthy donors (n=7) were analyzed by small RNA sequencing to define disease-associated tRF signatures. In vitro , primary chondrocytes derived from OA patients were treated with lipopolysaccharide (LPS) to mimic inflammatory environment of OA, used for small RNA sequencing (n=3) and validation analysis (n=6). Functional studies in C28/I2 chondrocytes included antisense oligonucleotide-mediated 3’tRF ^Asp(GTC) inhibition, AGO2-RNA immunoprecipitation (RIP), polysome profiling, stress granule (SG) immunofluorescence, and differential protein analysis. Computational target prediction and pathway enrichment were used to explore tRF-mediated regulatory networks. Results: Both OA cartilage and LPS-treated chondrocytes displayed upregulation of 3’tRF ^Asp(GTC) and 5’tRF ^Glu(CTC) , indicating a shared inflammatory tRF signature. Predicted targets of upregulated tRFs were enriched in stress-adaptive, proteostasis, and translational control pathways, whereas downregulated tRFs modulated mitochondrial processes. Silencing 3’tRF ^Asp(GTC) inhibited LPS-induced COX2 and MMP13 expression, prevented ER stress, and blocked SG assembly. RIP confirmed selective recruitment of 3’tRF ^Asp(GTC) into AGO2 complexes. Polysome profiling revealed association with 40S ribosomal subunit, mediating translational arrest and influencing selective mRNA expression. Conclusion: 3’tRF ^Asp(GTC) emerges as a regulator linking inflammation to translational control and SG dynamics in OA. tRFs thus could represent novel therapeutic targets in OA disease.

Human skeletal remains are a crucial source for understanding biocultural and evolutionary processes. Yet, their study and management are challenged by social, religious, and political factors, placing them in a ‘grey area’ within cultural heritage. Human skeletal collections often carry colonial legacies, raising ethical concerns and new challenges for research, curation, and public engagement in academic and museum institutions. In this context, digitization offers expanding opportunities for public exhibition and definition of human remains as part of our culture, while ensuring long-term preservation. Virtual approaches serve also as a useful tool to investigate human variability from evolutionary, bioarchaeological, and forensic perspectives. Moreover, digital access fosters interdisciplinary collaboration and research by enabling global scholarly engagement beyond physical limitations. Through the CHANGES project, we have initiated the digitization of the Documented Human Osteological Collections (DHOC) of the University of Bologna—one of Italy’s largest collections—making these resources available via the 3D data repository MorphoSource. This contribution provides updates on newly digitized material and reports on access requests received to date. We conclude by considering the emerging responsibilities of anthropologists in the use of virtual human skeletal collections, promoting best practices for the management of the anthropological digital twins.