Background: The contamination of ambulances with pathogenic agents represents a potential threat for the public health, not only for common pathogens but also for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The aim of this project was to exploits the germicidal effect of the UVC radiation at 254 nm to sanitize the patient’s compartment of ambulances with an advanced UltraViolet SANitizing System (UV-SAN) and assess its relevance for avoiding the spread of COVID-19 and other drug resistant pathogens. Methods: The system is equipped with UVC lamps that are activated when the ambulance compartment is empty and sanitize the environment in less than 15 min. An Ozone sensor continuously monitors the gas concentration, ensuring it does not exceed threshold value harmful for patients and operators’ health. The system is relying on GNSS data and a satellite communication link, which allow to monitor and record traceability (when, where and what) of all the sanitation operations performed. This information is real-time monitored from a dedicated web-application. Results: UVC irradiation efficiently reduced SARS-CoV-2 virus titer (>99.99%), on inanimate surfaces such as plastic, stainless steel or rubber, with doses ranging from 5.5 to 24.8 mJ/cm² and the UV-SAN system is effective against multi drug resistant (MDR) bacteria up to >99.99%, after 10 to 30 min of irradiation. Conclusions: UV-SAN can provide rapid, efficient and sustainable sanitization procedures of ambulances.

Integrase-defective lentiviral vectors (IDLVs) represent an attractive platform for vaccine development as a result of the ability to induce persistent humoral- and cellular-mediated immune responses against the encoded transgene. Compared with the parental integrating vector, the main advantages for using IDLV are the reduced hazard of insertional mutagenesis and the decreased risk for vector mobilization by wild-type viruses. Here we report on the development and use in the mouse immunogenicity model of simian immunodeficiency virus (SIV)-based IDLV containing a long deletion in the U3 region and with the 3′ polypurine tract (PPT) removed from the transfer vector for improving safety and/or efficacy. Results show that a safer extended deletion of U3 sequences did not modify integrase-mediated or -independent integration efficiency. Interestingly, 3′ PPT deletion impaired integrase-mediated integration but did not reduce illegitimate, integrase-independent integration efficiency, contrary to what was previously reported in the HIV system. Importantly, although the extended deletion in the U3 did not affect expression or immunogenicity from IDLV, deletion of 3′ PPT considerably reduced both expression and immunogenicity of IDLV.

Background: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has been added to the World Health Organization’s list as a high-priority pathogen for which new antibiotics are urgently needed. Herein, we investigated the association between resistance/virulence genes and high-risk CRPA clinical isolates by whole genome sequencing (WGS). Methods: Between 2019 and 2025, twenty-six CRPA strains from patients hospitalized in the “Renato Dulbecco” University Hospital were characterized. WGS analysis was performed using the next generation sequencing (NGS) technique. Multi-locus sequence typing (MLST) prediction was performed. Antibiotic resistance genes were detected using Antibiotic Resistance Gene-ANNOTation, Comprehensive Antibiotic Resistance Database, and ResFinder. Virulence genes were identified by the Virulence Factor Database. Results: The MLST analysis detected 14 different sequence types (ST). The 26 strains exhibited the same resistome profile: aac(3)-Ic, aphA15, catB7, catB10, cmlA, blaCARB, blaVIM-1, and tetG genes. The genes encoding enzymes involved in resistance to chloramphenicol and beta-lactams were found in all isolates using the three databases. Biofilm formation genes, metalloproteinase, chemotaxis, fimbriae, and pyoverdine were identified in all strains. Genes of the type III secretion system exoS, exoT, exoU, and exoY were found in 46.15%, 84.61%, 53.84%, and 84.61% of the strains, respectively. Conclusions: The analysis of the 26 clinical isolates showed high clonal heterogeneity, with a predominance of ST235, a high-risk clone associated with multiple resistances. Interestingly, cefiderocol resistance was carried by 4/8 isolates belonging to the ST235 strain. The surveillance based on resistome and virulome analysis could monitor the dynamic evolution of high priorityhigh-priority pathogens to guide clinical treatment and to adapt healthcare control measures, limiting their spread in the near future.

In chronic lymphocytic leukemia (CLL), natural killer (NK) cells show a dysfunctional phenotype that correlates with disease progression. Our aim was to restore NK cell functionality in CLL through a specifically targeted IL15-stimulating activity; IL15 targeting could, in fact, potentiate the activity of NK cells and reduce off-target effects. We designed and developed a cis-acting immunocytokine composed of an anti-CD56 single-chain Fragment variable (scFv) and IL15, labeled scFvB1IL15. scFvB1IL15 was tested in vitro on peripheral blood mononuclear cells (PBMCs) obtained from both different healthy donors (HDs) and CLL patients in order to evaluate its ability to target NK cells and enhance their activation and NK-mediated directed cytotoxicity. scFvB1IL15 specifically induced strong degranulation and cytokine and chemokine production in NK cells in both HD- and CLL patient-derived PBMC samples. Furthermore, compared to IL15 alone, it was able to induce higher levels of NKG2D- and NKp30-activating receptors and restore NK-mediated direct killing in the CLL patient-derived samples. The preliminary data presented in this work suggest that IL15’s targeting of NK cells via scFvB1 potentiates the effects of IL15 and that scFvB1IL15 can be a useful agent for overcoming NK functional gaps and contribute to NK-cell-based immunotherapies.

If the number of viral hepatitis infections is to be decreased worldwide, and the World Health Organization (WHO) elimination targets are to be achieved by 2030, this requires determining the burden of infection according to the WHO’s test-and-treat approach. In 2014, the introduction of Direct-Acting Antivirals (DAAs) revolutionized the management of Hepatitis C Virus (HCV); another improvement came in 2020, when the use of bulevirtide (BLV) was authorized as a treatment for chronic Hepatitis D Virus (HDV) infection, showing good efficacy. The present observational study was carried out between 2019 and 2024. The diagnosis of viral hepatitis was carried out by routine assays. HDV typing was performed by Sanger sequencing and phylogenetic analysis. Overall, the HCV antibody prevalence was 3.4% in the studied time span, and it was higher in males than in females (59% vs. 41%). In viremic patients, HCV1b (33%) and HCV2a/2c (25%) were the most common subtypes. The overall HCV viremic rate declined in 2022 (2.8%). Unlike HCV, 71.4% of HDV viremic patients were females, and they had a median age of 58 years. The viral load of HDV RNA ranged from 20 IU/mL to 8 million IU/mL. Viral genotypes were classified as HDV1c and HDV1e. In this study, we highlight the prevalence of HCV/HDV infections and their genotype evolution in Southern Italy, underscoring the urgent need to enhance screening and linkage to care. Finally, we quantify the burden of active infections in order to provide data from real-life settings, and we describe the virological status of people living with HCV or HBV/HDV, who may experience significant benefits in terms of liver-related mortality after DAA or BLV treatment.