Lorenza Sanfilippo

Oropouche virus (OROV) has re-emerged as a significant public health threat during the 2023–2025 outbreak, characterized by an unprecedented geographic expansion beyond endemic regions. The virus has been detected in various regions, including the Caribbean, as well as in Europe and the United States, among travelers from Central and South America. This epidemic was associated with the expansion of a novel reassortant strain, which may exhibit enhanced viral fitness and immune evasion properties. Although orthobunyaviruses are known to counteract interferon responses, the specific strategy adopted by OROV remains incompletely understood. In this study, we compared the in-vitro behavior of the prototype strain BeAn19991 and a 2024 isolate representing the reassortant virus responsible for the 2023–2025 epidemic (ISCDC-2024), focusing on innate immune response. We examined the sensitivity of the two strains to IFN-β antiviral activity and their ability to induce type I IFN in fibroblast. Furthermore, we characterized the innate immune response triggered by in-vitro infection of PBMCs from healthy donors. Although the two viral strains exhibited a similar plateau in viral RNA accumulation and comparable sensitivity to exogenous IFN-β, the IFN-β induction in fibroblasts was delayed, with higher levels induced by ISCDC-2024 than by BeAn19991. Moreover, the two strains displayed distinct innate immune profiles in PBMCs: BeAn19991 predominantly induced a robust antiviral IFN-driven response, whereas ISCDC-2024 infection was associated with an IL-6-dominated inflammatory pattern. Overall, these findings highlight distinct phenotypic features of the contemporary reassortant isolate and suggest strain-specific differences in the modulation of innate immunity, potentially reflecting distinct immune evasion strategies. If confirmed, this hypothesis could have significant implications for infection transmission, viral persistence and disease severity, highlighting the need for continuous surveillance and functional characterization of emerging OROV strains to drive public health interventions and antiviral countermeasures.

Background/Objectives: Strongyloides stercoralis is a soil-transmitted helminth that can cause severe disease in immunocompromised individuals. In non-endemic clinical settings, recommended diagnostic strategies rely on serology alone or in combination with a sensitive fecal test, maximizing sensitivity to avoid missed infections. The S. stercoralis VirClia^® IgG monotest (Vircell, Spain) is a novel automated chemiluminescent immunoassay (CLIA) that detects IgG antibodies against the recombinant NIE and SsIR antigens. This study evaluated its diagnostic accuracy (sensitivity and specificity) in a non-endemic clinical setting. Methods: In this monocentric retrospective study, 245 serum samples from the Tropica Biobank of a referral centre for parasitic diseases in Italy were analyzed. Samples originated from individuals from Africa (n = 85), America (n = 31), Asia (n = 53), and Europe (n = 76). Diagnostic accuracy was estimated using Bayesian latent class analysis based on a model including five routinely used diagnostic tests. Results: The S. stercoralis VirClia^® IgG monotest showed a sensitivity of 95.0% (95% credible interval [CrI]: 90.1–98.2%) and a specificity of 90.8% (95% CrI: 85.1–96.0%) for the tested population. Conclusions: In immunocompetent individuals with a risk of previous exposure to S. stercoralis, the S. stercoralis VirClia^® IgG monotest showed high diagnostic accuracy. Its high sensitivity supports current recommendations favouring highly sensitive serological assays for screening and diagnosis in non-endemic clinical settings.

Introduction. The SPIRIT (Standard Protocol Items: Recommendations for Interventional Trials) Statement has long been the international reference for the content of clinical trial protocols, ensuring transparency, methodological rigour and ethical soundness [1]. The recent release of SPIRIT 2025 has updated and expanded its recommendations [2]. Nonetheless, numerous studies continue to document suboptimal adherence, particularly regarding study design and statistical analysis. Assessing real-world adherence helps identify recurrent shortcomings and areas requiring targeted support or training. Objectives. In a previous single-centre study we assessed whether it is possible to predict which protocols show high adherence to the methodological SPIRIT items, identifying study-level characteristics associated with quality. The main limitation was the single-centre nature of the data. The present work extends the analysis to a multicentre design, combining protocols from two Italian research centres: Fondazione IRCCS Policlinico San Matteo (Pavia) and IRCCS Sacro Cuore Don Calabria (Negrar di Valpolicella, Verona), aiming to confirm the predictors of adherence and improve their generalisability. Methods. Design and methodological features of no-profit interventional protocols submitted between 2021 and 2025 to the reference Ethics Committees of the two centres were collected and recorded in a centralised REDCap (Research Electronic Data Capture) database. Adherence was assessed on the SPIRIT 2013 items concerning methods, design, data collection and management, and analysis (items 9-21b); the 2013 version was used because all protocols predate SPIRIT 2025. Each item was scored as fulfilled or not, and an individual score was computed as the number of satisfied items, dichotomised at the median into good and poor adherence. For the predictive objective, machine learning algorithms (Random Forest, eXtreme Gradient Boosting, Boosted Logistic Regression) were applied to a pool of candidate predictors; performance was evaluated through accuracy, area under the ROC curve (AUC) and F1 score. Variable importance was then summarised and the most influential predictors were entered into a multivariable logistic regression model, reporting odds ratios and 95% confidence intervals. Covariates included sponsorship, methodological features, submission timing and thematic area. Analyses were performed using Stata 19 and R 4.4.3. Results. Data collection and harmonisation across the two centres are currently ongoing. The IRCCS Sacro Cuore Don Calabria contributed 107 no-profit interventional protocols submitted to its reference Ethics Committee between 2021 and 2025. Combined with the 132 protocols of the previous single-centre study, the pooled cohort reaches 239 protocols. This larger and more heterogeneous sample is expected to produce more stable predictive models and to enable a formal assessment of the reproducibility of the predictors identified in the single-centre analysis. We expect to confirm the availability of statistical and methodological support as a factor associated with good adherence, while the relative contribution of the remaining candidate predictors (sponsorship, methodological features, submission timing and thematic area) will be clarified in the pooled cohort. The machine learning algorithms are expected to discriminate good from poor adherence and to converge on a consistent subset of influential predictors, which the multivariable logistic regression should retain as independent. The final estimates will be presented. Conclusions. The multicentre extension addresses the main limitation of the previous work, strengthening the external validity of the findings. By integrating conventional statistical approaches with machine learning models, the study aims to clarify which protocol features predict high adherence to SPIRIT and to confirm the role of multidisciplinary teams, including biostatistics, in improving methodological quality. The findings are expected to provide evidence to inform training strategies aimed at improving the quality of clinical trial protocols.

Purpose: to outline the role of CT in pre- and post-treatment evaluation in the case of lumbar endoscopic foraminotomy. Methods: This prospective study, conducted between September 2020 and January 2024, included consecutive patients with clinical symptoms of lumbar sciatica/lumbalgia/lombo-cruralgia/lower limb peripheral neuropathy. Pre- and post-foraminotomy CT imaging was used to assess the foraminal diameters (cranio-caudal, transverse and free hand ROI area) before and after the treatment. Two independent blinded readers assessed the CT randomly. VAS pain scale and the measurements of each foramen were compared before and after treatment. Interobserver agreement was assessed using the Intraclass Correlation Coefficient (ICC). Results: A total of 47 participants were enrolled, with 53 intervertebral levels analyzed. The mean VAS value decreased from 9.17 in the preoperative period to 0.66 at the one-month postoperative follow-up. The clinical response was associated with statistically significant changes in the cranio-caudal and transverse diameters, as well as the area of the treated neuroforamina (p-values < 0.05). Inter-rater reliability between the two operators ranged from 0.75 to 0.90. Conclusions: CT can demonstrate a significant enlargement of the neuroforaminal diameters after the endoscopic foraminotomy, with good correlation with clinical improvement.

Dual-energy CT helped to accurately diagnose bone stress fractures and stress reactions of the lower limb (leg, ankle, and foot) using MRI as the reference standard.