Ospedale Pediatrico Bambino Gesù

1) to evaluate the feasibility of a pilot trial on the effectiveness of a pediatric intensive care unit (PICU) diary intervention; 2) to explore group differences in parent's and child's psychological outcomes and children's behaviors after discharge from PICU.

A non-blinded, single-centered, pilot randomized controlled trial (RCT) was performed in two PICUs at a tertiary care children's hospital in Italy. Children admitted to the PICU, <18 years old, sedated and intubated for ≥ 48 h were eligible. The exclusion criteria included severe disability (Pediatric Overall Performance Category > 3), low Italian communication proficiency, and no life expectancy. The intervention was the PICU narrative diary.

A total of 119 patients were prospectively enrolled from June 2021 to June 2024, of which 60 received a PICU diary, and 59 did not. The composite feasibility outcomes, including eligibility, recruitment, retention, attrition, and protocol adherence met 6 of the 8 criteria. At 1 and 3 months after PICU discharge Post Traumatic Stress Disease (PTSD), anxiety, depression in parents and children, and children's behaviors were measured with validated scales. At the one-month follow-up, parents in the control group had a higher PTSD (21 % vs 11 % p = 0.1) and depression (28 % vs 25 % p = 0.7) and lower anxiety (54 % vs 60 % p = 0.5) compared to the intervention group. At 3 months, lower levels were observed across all outcomes. Parent satisfaction with the intervention was high. The small sample size and lack of blinding were the main study limitations.

The trial was feasible and acceptable. No significant differences in parents' and children' psychological outcomes were found among the study groups.

PICU diaries could be a valuable support tool for parents and children admitted to PICU. Further evidence through larger RCTs is needed to understand the effect of post-PICU discharge psychological outcomes on children and their parents.

Gliomas are aggressive brain tumors whose infiltrative growth is mediated by intercellular crosstalk. Exosomes, small extracellular vesicles, play a key role in cell-cell communication but are difficult to visualize using conventional microscopy. Performing immunostaining for CD63, a known exosome marker, and using STED microscopy, we demonstrate exosome secretion in primary glioma cells. Applying mathematical deconvolution, we enhance the contrast and resolution for in-depth analysis of STED images. We identify CD63-positive cellular footprints and exosome deposits in the extracellular space. Quantitative analysis shows CD63-positive exosomes ranging 36.55-157.06 nm in size. CD63/actin co-staining demonstrates different actin polymerization states associated with exosomes. In conclusion, STED microscopy coupled with immunostaining allows exosome primary characterization at the single-vesicle level in the cellular spatial context.