Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis-An experimental study.

BACKGROUND: The development of next-generation sequencing approaches has accelerated the diagnostic process, although at present, there is a lack of a clear consensus on efficient management of human samples for downstream applications. This study aims to investigate timeframe (in terms of short preservation), temperature, and additional preservation procedures (i.e., freeze and thaw cycles) for human biospecimens to implement the reliability and reproducibility of molecular investigations. METHODS: Overall, 45 whole peripheral bloods, 22 peripheral blood mononuclear cells samples, 15 saliva, and 15 buccal swab biospecimens (through the extracted DNA) were investigated, assessing yield, integrity, amplifiability, and sizing accuracy via the most common molecular techniques. RESULTS: Based on the overall evaluation criteria, the results indicate that DNA extracted from all samples, shortly preserved, have suitable quality and reliable reproducibility to be used in diagnostic activities and biomedical research, even if DNA from peripheral blood mononuclear cells is more affected by the experimental conditions. CONCLUSION: Our findings confirm the reliability of peripheral blood samples in almost all the experimental conditions. Saliva and buccal swabs are efficient almost as well, while peripheral blood mononuclear cells, albeit remain a primary source of DNA for molecular screenings, represent a less efficient source.

Osteogenesis imperfecta: a cross-sectional study of skeletal and extraskeletal features in a large cohort of Italian patients.

Introduction: The present study aims to describe a large cohort of Italian patients affected by osteogenesis imperfecta, providing a picture of the clinical bony and non-bony features and the molecular background to improve knowledge of the disease to inform appropriate management in clinical practice. Methods: A total of 568 subjects (from 446 unrelated Italian families) affected by osteogenesis imperfecta who received outpatient care at Istituto Ortopedico Rizzoli from 2006 to 2021 were considered in the present study. Results: Skeletal and extraskeletal features were analyzed showing a lower height (mean z-scores equal to -1.54 for male patients and -1.47 for female patients) compared with the general Italian population. Half of the patient population showed one or more deformities, and most of the patients had suffered a relatively low number of fractures (<10). An alteration in the sclera color was identified in 447 patients. Similarly, several extraskeletal features, like deafness, dental abnormalities, and cardiac problems, were investigated. Additionally, inheritance and genetic background were evaluated, showing that most of the patients have a positive family history and the majority of pathogenic variants detected were on collagen genes, as per literature. Conclusion: This study supports the definition of a clear picture of the heterogeneous clinical manifestations leading to variable severity in terms of skeletal and extra-skeletal traits and of the genetic background of an Italian population of osteogenesis imperfecta patients. In this perspective, this clearly highlights the crucial role of standardized and structured collection of high-quality data in disease registries particularly in rare disease scenarios, helping clinicians in disease monitoring and follow-up to improve clinical practice.

Effectiveness of a "Spring Pillow" Versus Education in Chronic Nonspecific Neck Pain: A Randomized Controlled Trial.

BACKGROUND: Different types of pillows have been proposed for neck pain, but no previous randomized controlled trial has investigated the effectiveness of a "spring pillow" for adults with chronic nonspecific neck pain. OBJECTIVE: We evaluated the effectiveness of using a pillow made from viscoelastic polyurethane and 60 independent springs compared with an educational intervention in individuals with chronic nonspecific neck pain. DESIGN: This was a randomized controlled trial with crossover study design. SETTING: The setting was the Occupational Medicine Unit, University Hospital, Bologna (Italy). PARTICIPANTS: We recruited 70 adults with chronic nonspecific neck pain, of whom 64 completed the trial. INTERVENTION: Participants were randomly assigned to 2 groups. One group used the spring pillow for 4 weeks, and the other group followed educational advice for 4 weeks while continuing to use their own pillows. After 4 weeks of treatment and 4 weeks of washout, groups were crossed over. Pain perceived in the neck, thoracic, and shoulder areas and headache were the primary outcome measures. In addition, disability, sleep quality, subjective improvement, and pillow comfort were assessed. Measures were captured at pretreatment, after 4 weeks, after the 4-week washout period, and 4 weeks after crossover. The mean differences (MD) in outcomes between groups were assessed. RESULTS: Treatment with the spring pillow appeared to reduce neck pain (MD = -8.7; 95% confidence interval [CI] = -14.7 to -2.6), thoracic pain (MD = -8.4; 95% CI = -15.2 to -1.5), and headache (MD = -16.0; 95% CI = -23.2 to -8.7). Reductions in shoulder pain were not statistically significant between groups (MD = -6.9; 95% CI = -14.1-0.3). Neither the crossover sequence nor the period (first vs second intervention administration) significantly affected the results. LIMITATIONS: Education may not have been the best comparator for the spring pillow; drug consumption, actual pillow use, and the implementation of the educational suggestions as prescribed were not controlled. CONCLUSIONS: Use of the spring pillow in this study was more effective than an educational intervention for improving cervical, thoracic, and head pain. Whether a spring pillow is more effective than other ergonomic pillows remains to be tested.