25-Hydroxycholesterol (25-HC) and 25R,26-hydroxycholesterol (25R,26-HC) are two endogenous oxysterols endowed with broad-spectrum antiviral activity. They hamper viral replication by targeting oxysterol-binding protein (OSBP), a host lipid transporter which plays a critical role in the replicative cycle of several viruses. Recently, we have reported the remarkable antiviral activity against herpes simplex virus 2 (HSV-2) of N,N-dimethyl-3β-hydroxychol-5-en-24-amide (PFM067, 3), identified by the screening of our synthetic oxysterol library. Further development of antiviral cholenamide-based fluorescent probes allowed us to support the hypothesis that OSBP was involved in the antiviral mechanism of action of the parent PFM067. Herein, we report the first study aimed at the definition of structure-antiviral activity relationships for PFM067 (3); successfully, we identified 24-(morpholin-4-yl)-3β-hydroxychol-5-en-24-one (8d), as highly potent, nanomolar inhibitor of HSV-2 replication, endowed with promising selectivity index. Moreover, we demonstrated that OSBP was the molecular target underlying the anti-HSV-2 activity of PFM067 (3) and its analogues by the development of a microscale thermophoresis binding assay. In-depth experiments performed using an OSBP-silenced cell line allowed us to disclose the role played by OSBP in the replicative cycle of HSV-2. Finally, molecular modelling studies evidenced a peculiar orientation of the active compound 8d within the binding site of OSBP respect to that stroked by either the endogenous ligand or an inactive compound.

Acute respiratory infections are the most common cause of acute illness globally and have a severe impact on human health and productivity. There is robust evidence of airborne transmission of many respiratory viruses via direct or indirect contact with droplets and aerosol produced by infected individuals, and this transmission route becomes crucial in crowded indoor spaces. Heating, ventilating and air conditioning (HVAC) systems filters can reduce the concentration of virus-carrying droplets, but HVAC systems able to directly inactivate bacteria and viruses are highly desirable to preserve safe indoor air.The aim of the present work was to assess the antibacterial and antiviral properties of silver nanoclusters/silica or zirconia composite coatings deposited onto polymeric air filters against Staphylococcus epidermidis, Staphylococcus aureus and a panel of representative members of human respiratory viruses, such as human coronavirus OC43 (HCoV-OC43), human rhinovirus A1 (HRV-A1), influenza virus type A (IFVA-H3N2) and adenovirus type -5 (AdV-5) according to standard protocols ISO.Results evidenced that both coatings showed a significant antimicrobial and antiviral activity at variable extent and a good cytocompatibility on all the examined cell lines, demonstrating a broad-spectrum action against selected bacteria and respiratory viruses.

Pathogenic air and water contaminations present major health issues nowadays. Filter-based protective devices have been commonly utilized to capture airborne or waterborne microorganisms and viruses in order to reduce or inhibit the transmission of infectious diseases. Some approaches to extend the functionality of filtering devices and to further enhance their capability to inactive pathogens include the application of antimicrobial coatings on the filter surface. In this regard, the current work aims to develop innovative coatings with antiviral and antibacterial properties composed of a chitosan matrix incorporating bioactive glass (BG) nanoparticles or silver nitrate (AgNO3) on metallic filters by using electrophoretic deposition. The so-produced chitosan/BG- and chitosan/AgNO3-coated filters were characterized in terms of their morphology, chemical composition, filtering capability, antibacterial properties against Staphylococcus aureus, and antiviral activity against coronavirus OC43, rhinovirus, adenovirus, and influenza virus. The results confirmed that chitosan/AgNO3 and chitosan/BG coatings exhibited no blockage of the filter pores. The bubble point values for both coating types were similar to the measurements of uncoated filters, suggesting intact filtration capability after the application of the coatings. The antibacterial assay showed the antibacterial activity of >99% for both chitosan/AgNO3 and chitosan/BG coatings against S. aureus. Finally, chitosan/AgNO3-coated filters showed antiviral activity against three out of the four tested viruses (HCoV-OC43, HRV-A1, and IFV-AH3N2) and exerted a partial antiviral action against AdV-5. On the other hand, chitosan/BG-coated filters were active against all tested respiratory viruses. The outcomes of this study evidenced that the developed multimaterial coatings could serve as a suitable platform to produce antimicrobial filtering systems for a broad-spectrum application.

The increasing demand for sustainable chemical processes has driven the search for renewable feedstocks, environmentally benign catalysts, and energy-efficient methodologies. In this context, we report a solvent-free protocol for the Claisen-Schmidt condensation between biomass-derived furanic aldehydes and acetophenone. Commercial magnesium oxide (MgO) was employed as a recyclable heterogeneous catalyst and subjected to physicochemical characterization and recycling tests to evaluate its stability and reusability. Microwave (MW) irradiation was integrated to ensure rapid and homogeneous heating, leading to enhanced reaction efficiency and reduced processing times. The sustainability of the proposed approach was preliminarily assessed through green chemistry metrics, which confirmed its advantages over conventional methods. The obtained furano-chalcones were investigated for their antiviral potential in in vitro cell-based models against common human pathogenic viruses, such as human herpes simplex virus, Zika virus, rhinovirus and influenza virus. Their activity was specifically targeted against HSV type 2, highlighting their relevance as pharmacologically active scaffolds and warranting further optimization and investigation. Overall, this work combines renewable resources, recyclable catalysis, and energy-efficient techniques, offering a greener and versatile strategy for the synthesis of high-value bioactive compounds.

Varicella-zoster Virus (VZV) is a relevant pathogen belonging to the herpesviridiae family. Primary VZV infection causes chickenpox, and results in latent infection of sensory ganglia. Later in life, VZV can reactivate causing herpes zoster (HZ), which can be associated with severe complications in immunocompromised individuals. Currently, the available antivirals used to treat VZV infection target the DNA replication stage; however, resistance to these drugs has been reported in both immunocompromised and immunocompetent patients. For this reason, the identification of new antiviral molecules against VZV infection is a priority. Recently our research group demonstrated that the endogenous oxysterol 25R,26-hydroxycholesterol (25R,26OHC, more commonly named 27-hydroxycholesterol) and an oxysterol synthetic analog named PFM067 inhibit herpes simplex virus (HSV) replication. In this study we explored the antiviral activity of 25-hydroxycholesterol (25OHC), 25R,26OHC, and PFM067 against VZV. We demonstrated that 25R,26OHC and PFM067 exert antiviral activity against VZV with an EC50 in the low micromolar range and are able to significantly reduce the area of the viral plaques. Moreover, 25R,26OHC and PFM067 can inhibit the egress of viral glycoprotein gE from the cis-Golgi compartment, similarly to what demonstrated by our group for HSV-2. Additionally, we show that 25R,26OHC and PFM067 act synergistically when used in combination with acyclovir (ACV). The promising antiviral activity of 25R,26OHC and PFM067, along with their different mechanism of action compared to ACV, makes these molecules suitable candidates for further investigation of the molecular target of oxysterols.

Broadly acting antivirals are needed to complement vaccines in present and future pandemics and outbreaks as safe and sustainable tools for combating virus infections. Thus, the current study aims to investigate the broad-spectrum antiviral activity of ferments of selected strains belonging to the Ganoderma lucidum complex isolated from Finland and investigate their mechanism of action. Cytopathic effect inhibition assay and endpoint assay were used to determine the antiviral activity. The antibacterial activity ws examined using recombinant biosensor strains. Mechanism of action studies included time of addition assay, transmission electron microscopy, confocal microscopy, sucrose gradient separation assay, and thermal assay. The metabolite composition of the ferments was studied using UHPLC-HR-MS/MS. Ferments showed antiviral efficacy against non-enveloped enteroviruses, already at room temperature and within one minute. Broad antimicrobial activity was demonstrated with non-enveloped rotaviruses, enveloped coronaviruses, zika viruses, and gram-positive and gram-negative bacteria. Ferments also directly affected the viruses and caused clustering of the virus particles. Additionally, treatment of enteroviruses with ferments caused strong stabilization of the virus capsid, thus preventing their genome release. UHPLC-HR-MS/MS analysis of the ferments verified the presence of several terpenoid compounds. The results show great promise for the future use of ferments from the Ganoderma lucidum complex in combating microbial infections for various applications.

Cell-based phenotypic screening of a privileged in-house library composed of pyridobenzothiazolone (PBTZ) analogues was conducted against representative viruses responsible for common respiratory tract infections in humans, i.e., respiratory syncytial virus (RSV), human coronavirus type OC43 (HCoV-OC43), and influenza virus type A (IFV-A). We identified a compound with broad-spectrum inhibitory activity against multiple strains of RSV, HCoV, and IFV, with EC50 values in the low micromolar range and cell-independent activity. Its antiviral activity and cytocompatibility were confirmed in a fully differentiated 3D model of the bronchial epithelium mimicking the in vivo setting. The hit compound enters cells and localizes homogeneously in the cytosol, inhibiting replicative phases in a virus-specific manner. Overall, the selected PBTZ represents a good starting point for further preclinical development as a broad-spectrum antiviral agent that could address the continuous threat of new emerging pathogens and the rising issue of antiviral resistance.

Marine microplastics (MPs) represent a novel ecological niche, populated by fungi with high potential for pharmaceutical discovery. This study explores the bioactivity of fungal strains isolated from MPs in Mediterranean sediments, focusing on their osteogenic and antiviral activities. Crude extracts prepared via solid-state and submerged-state fermentation were tested for their effects on extracellular matrix mineralization in vitro and bone growth in zebrafish larvae, and for their activity against the respiratory syncytial virus (RSV) and herpes simplex virus type 2 (HSV-2). Several extracts exhibited significant mineralogenic and osteogenic activities, with Aspergillus jensenii MUT6581 and Cladosporium halotolerans MUT6558 being the most performing ones. Antiviral assays identified extracts from A. jensenii MUT6581 and Bjerkandera adusta MUT6589 as effective against RSV and HSV-2 at different extents, with no cytotoxic effect. Although chemical profiling of A. jensenii MUT6581 extract led to the isolation of decumbenones A and B, they did not reproduce the observed bioactivities, suggesting the involvement of other active compounds or synergistic effects. These results highlight the plastisphere as a valuable resource for novel bioactive compounds and suggest the need for further fractionation and characterization to identify the molecules responsible for these promising activities.

Sexually transmitted infections (STIs) remain a major global health challenge, highlighting the urgent need for effective and user-friendly vaginal prevention strategies. This study presents a novel composite system for vaginal application, consisting of mucoadhesive electrospun nanofibres with inherent antiviral potential embedded within a pH-responsive film. The film is designed to preserve the integrity of the nanofibres in the acidic vaginal environment and to dissolve rapidly upon contact with seminal fluid - released during sexual intercourse -, triggering nanofibre hydration and interaction with the mucosal surface. Electrospinning successfully produced uniform and defect-free nanofibres consisting of polyvinyl alcohol (PVA) blended with either κ- or ι-carrageenans (CAR), sulphated polysaccharides known for their mucoadhesive, gelling and intrinsic antiviral properties. Different solutions containing Eudragit® polymers (EL100 or EL100-55) and plasticisers (polyethylene glycol or glycerol) were prepared and cast to identify the most suitable composition for developing the composite system. Solutions capable of forming films with optimal mechanical flexibility and rapid solubility under mildly alkaline conditions were selected. The composite system was fabricated by embedding nanofibres between two partially dried layers of the selected pH-responsive solutions, forming a uniform composite structure that ensured complete fibre incorporation. The outer film effectively protected the nanofibrous core in acidic environments; upon pH increase (pH ∼7.5), the film rapidly dissolved, allowing the nanofibres to hydrate and form a cohesive, strongly mucoadhesive hydrogel, potentially enhancing their retention within the vaginal cavity. Overall, the composite system exhibited good structural integrity, pH-responsiveness, biocompatibility and antiviral potential, offering a promising, non-hormonal strategy for on-demand STI prevention.

During the SARS-CoV-2 pandemic, many countries established wastewater (WW) surveillance to objectively monitor the level of infection within the population. As new variants continue to emerge, it has become clear that WW surveillance is an essential tool for the early detection of variants. The EU Commission published a recommendation suggesting an approach to establish surveillance of SARS-CoV-2 and its variants in WW, besides specifying the methodology for WW concentration and RNA extraction. Therefore, different groups have approached the issue with different strategies, mainly focusing on WW concentration methods, but only a few groups highlighted the importance of prefiltering WW samples and/or purification of RNA samples. Aiming to obtain high-quality sequencing data allowing variants detection, we compared four experimental conditions generated from the treatment of: i) WW samples by WW filtration and ii) the extracted RNA by DNase treatment, purification and concentration of the extracted RNA. To evaluate the best condition, the results were assessed by focusing on several sequencing parameters, as the outcome of SARS-CoV-2 sequencing from WW is crucial for variant detection. Overall, the best sequencing result was obtained by filtering the WW sample. Moreover, the present study provides an overview of some sequencing parameters to consider when optimizing a method for monitoring SARS-CoV-2 variants from WW samples, which can also be applied to any sample preparation methodology.

Oxysterols (OSs) represent a large family of cholesterol-derived molecules, involved in several physiological and pathological processes. Recently, we reported the remarkable antiviral activity against herpes simplex virus 2 (HSV-2) infection of three cholenamide or homocholenamide derivatives, namely PFM067, PFM064, and PFM069, identified by the screening of an in-house library of OS derivatives. With the aim to shed light on the antiviral mechanism of action of this class of molecules, we assumed to exploit the use of cholenamide-based fluorescent probes.Herein, we report that PFM120 and PFM124, two fluorescent tagged version of PFM067 maintain the same antiviral properties against HSV-2 as the parent compound and localize intracellularly inside the endoplasmic reticulum and the cis-Golgi network. Moreover, we also demonstrate that both tagged molecules co-localize with oxysterol-binding protein (OSBP) and are able to induce its re-localization. Finally, we report that PFM120 and PFM124 are endowed with antiviral activity against another OSBP-dependent viral pathogen, i.e. the human rhinovirus (HRV), different in structure and replication strategy from HSV-2.Taken together, these results candidate PFM120 and PFM124 as useful tools to investigate the actual mechanism of action and molecular target(s) of cholenamide-based antivirals and provide a proof of principle to explore them as a promising broad-spectrum class of antiviral agents.

After the end of the COVID-19 public health emergency, we analysed the relationship between Systemic Lupus Erythematosous (SLE) and COVID-19 from the virologist's perspective based on recent findings. SLE and COVID-19 co-morbidity present unique challenges, as individuals with SLE may be at increased risk for severe COVID-19 illness due to immune system abnormalities and ongoing therapies. Effective management of both diseases requires careful monitoring, adherence to vaccination programs, preventive measures and approved and patient-tailored therapies. This review covers various aspects, including the clinical outcome of SLE patients infected by SARS-CoV-2, the impact of this infection on SLE onset or flare-ups and the benefits of vaccination for this population. Furthermore, this review presents the most recent recommendations on clinical management of COVID-19 in rheumatic patients, including those with SLE, discussing the currently available therapeutic options. Finally, we explore the most effective tools for SARS-CoV-2 diagnosis in autoimmune conditions and examine prognostic biomarkers in COVID-19 rheumatic patients with potential implications on their clinical oversight. By adopting a comprehensive approach, we address these complexities from the virologist's perspective, aiming to improve health care for this vulnerable population.

Background While the prevalence of antiphospholipid antibodies (aPL) in venous and arterial thrombotic events had already been estimated by previous studies, the prevalence of aPL in subjects with Thrombotic Microangiopathy (TMA) is still not fully elucidated. Thus, we conducted a systematic review to estimate the frequency of aPL in subjects with biopsy-proven renal TMA. Methods We conducted in the PubMed database a search for English-language studies investigating the presence of aPL in subjects with biopsy-proven renal TMA from January 1985 to December 2022. Keywords used in the search included: ‘antiphospholipid syndrome’, ‘antiphospholipid antibodies’ and ‘thrombotic microangiopathy’. Cohorts of HUS patients were excluded due to the risk of over-estimating the prevalence of aPL in these populations. The median frequency for positive aPL including anticardiolipin antibodies (aCL), antibodies against β2-glycoprotein-I (anti-β2GPI) and lupus anticoagulant (LA) was then calculated. Results 522 articles were identified through the literature search. Six studies, assessing the prevalence of aPL in 211 subjects with renal TMA, were retrieved. The overall aPL prevalence was estimated as 24.4% (range 22–56). The estimated prevalence of aCL (IgG/IgM), anti-β2GPI, (IgG/IgM) and LA was 4.0% (range 3–27), 4.0% (range 3–16) and 18.9% (range 13–25), respectively. APS was diagnosed in 16.3% (range 11–29) of the patients. Of note, a high level of heterogeneity was observed when comparing the reported aPL profiles for each study. Conclusions This comprehensive systematic analysis of studies investigating the prevalence of aPL in renal TMA showed that, despite the high heterogeneity of the included studies, aPL are present in about one case out of four renal-TMA cases.

Abstract Introduction. The bacterial genotoxin colibactin is enriched in colorectal cancer (CRC) and promotes the accumulation of mutations that drive tumorigenesis. However, systematic assessment of its impact on DNA damage response is lacking and the effect of colibactin exposure on response to other genotoxic agents (such as chemotherapy) is missing. Materials and methods: We implemented an in vitro bacteria-coculture system to assess the effect of colibactin on a representative subset of 40 molecularly and pharmacologically annotated CRC cell lines and in a panel of isogenic DDR KO cell lines we generated. We further validated our results in patient-derived organoids. Finally, we recapitulated prolonged exposure to colibactin occurring during tumorigenesis by chronically infecting sensitive cells until the emergence of a tolerant phenotype. Results: We found that different cell lines display specific sensitivity to colibactin’s genotoxic stress: while colibactin-tolerant cells are capable of quickly and efficiently repairing colibactin-induced DNA damage, sensitive cells lack this ability. Moreover, we found that homologous recombination (HR) proficiency discriminates colibactin-tolerant cells, which display higher levels of RAD51 foci (as marker of activation of HR) compared to sensitive cells upon infection with colibactin. Screening of isogenic DDR KO cell lines revealed that genetic inactivation of the intertwined pathways of HR (through KO of ATM) and replication stress (RS) response (through KO of ATRIP) significantly sensitized cells to colibactin. In addition, we found that restoration of HR activity was sufficient to induce a colibactin-tolerant phenotype in initially sensitive cell lines. Notably, thanks to a previous effort of pharmacological characterization of CRC cell lines in our lab, we found a significant correlation between sensitivity to colibactin and irinotecan active metabolite SN38, but not oxaliplatin. We validated the same correlation in patient-derived organoids annotated for response to SN38. While colibactin, SN38 and oxaliplatin all induced RS in treated cells, we found that colibactin and SN38 showed a similar DNA damage response which involved activation of ATM. Finally, chronic re-infection of sensitive, HR-deficient CRC cells with colibactin selected a tolerant phenotype characterized by restoration of HR activity. Of translational relevance, colibactin-tolerant derivative cells acquired cross-resistance to SN38 and PARP inhibitor olaparib but not to oxaliplatin. Conclusion: Our results shed novel insight into colibactin’s genotoxic mechanism and support a model in which colibactin both promotes tumorigenesis and acts as an evolutionary bottleneck which selects HR proficient CRC cells. Furthermore, our study provides preclinical evidence on colibactin’s role in promoting chemoresistance in colorectal cancer. Citation Format: Alberto Sogari, Emanuele Rovera, Nicole Megan Reilly, Simona Lamba, Erika Durinikova, Annalisa Lorenzato, Marco Avolio, Eleonora Piumatti, Mariangela Russo, Sabrina Arena, Livio Trusolino, Manuela Donalisio, Federica Di Nicolantonio, David Lembo, Alberto Bardelli. Tolerance to colibactin correlates with response to chemotherapeutic agents in colorectal cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: DNA Damage Repair: From Basic Science to Future Clinical Application; 2024 Jan 9-11; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2024;84(1 Suppl):Abstract nr B005.

Coronaviruses (CoVs) share key genomic elements critical for viral replication, suggesting the feasibility of developing therapeutics with efficacy across different viruses. In a previous work, we demonstrated the antiviral activity of the antipsychotic drug lurasidone against both SARS-CoV-2 and HCoV-OC43. In this study, our investigations on the mechanism of action of lurasidone suggested that the drug exhibits antiviral activity by targeting the papain-like protease (PL-Pro) of both viruses, and the Spike protein of SARS-CoV-2, thereby hampering both the entry and the viral replication. In vitro assays demonstrate that lurasidone significantly reduces viral load in infected cells, showing that the drug is a promising candidate for further development as a dual-action antiviral, offering a potential new strategy in the fight against COVID-19 and other coronavirus-related diseases.

Human respiratory viruses have an enormous impact on national health systems, societies, and economy due to the rapid airborne transmission and epidemic spread of such pathogens, while effective specific antiviral drugs to counteract infections are still lacking. Here, we identified two Keggin-type polyoxometalates (POMs), [TiW11CoO40]8- (TiW11Co) and [Ti2PW10O40]7- (Ti2PW10), endowed with broad-spectrum activity against enveloped and non-enveloped human respiratory viruses, i.e., coronavirus (HCoV-OC43), rhinovirus (HRV-A1), respiratory syncytial virus (RSV-A2), and adenovirus (AdV-5). Ti2PW10 showed highly favorable selectivity indexes against all tested viruses (SIs >700), and its antiviral potential was further investigated against human coronaviruses and rhinoviruses. This POM was found to inhibit replication of multiple HCoV and HRV strains, in different cell systems. Ti2PW10 did not affect virus binding or intracellular viral replication, but selectively inhibited the viral entry. Serial passaging of virus in presence of the POM revealed a high barrier to development of Ti2PW10-resistant variants of HRV-A1 or HCoV-OC43. Moreover, Ti2PW10 was able to inhibit HRV-A1 production in a 3D model of the human nasal epithelium and, importantly, the antiviral treatment did not determine cytotoxicity or tissue damage. A mucoadhesive thermosensitive in situ hydrogel formulation for nasal delivery was also developed for Ti2PW10. Overall, good biocompatibility on cell lines and human nasal epithelia, broad-spectrum activity, and absence of antiviral resistance development reveal the potential of Ti2PW10 as an antiviral candidate for the development of a treatment of acute respiratory viral diseases, warranting further studies to identify the specific target/s of the polyanion and assess its clinical potential.

The bacterial genotoxin colibactin promotes colorectal cancer (CRC) tumorigenesis, but systematic assessment of its impact on DNA repair is lacking, and its effect on response to DNA-damaging chemotherapeutics is unknown. We find that CRC cell lines display differential response to colibactin on the basis of homologous recombination (HR) proficiency. Sensitivity to colibactin is induced by inhibition of ATM, which regulates DNA double-strand break repair, and blunted by HR reconstitution. Conversely, CRC cells chronically infected with colibactin develop a tolerant phenotype characterized by restored HR activity. Notably, sensitivity to colibactin correlates with response to irinotecan active metabolite SN38, in both cell lines and patient-derived organoids. Moreover, CRC cells that acquire colibactin tolerance develop cross-resistance to SN38, and a trend toward poorer response to irinotecan is observed in a retrospective cohort of CRCs harboring colibactin genomic island. Our results shed insight into colibactin activity and provide translational evidence on its chemoresistance-promoting role in CRC.

Infectious and toxicological risks are the main potential hazards that operators of Human Milk Banks (HMBs) encounter and must eliminate. HMBs are trying to implement procedures that allow to manage and sanitize human milk without altering significantly its nutritional and biologically protective components, obtaining a product characterized by a valid balance between safety and biological quality. The history of human milk processing is linked to the origins of HMBs themselves. And although other forms of sterilization were used originally, pasteurization soon became the recognized most effective means for sanitizing milk: all the milk that arrives at the HMB must be pasteurized. Holder pasteurization (HoP) is the most used methodology, and it is performed using low temperature and long time (+62.5°C for 30 min). With HoP some bioactive milk components are lost to varying degrees, but many other precious bioactive compounds are completely or partially preserved. To improve the quality of human milk processed by HMBs, maintaining in the meantime the same microbiological safety offered by HoP, new technologies are under evaluation. At present, High-Temperature Short-Time pasteurization (HTST) and High-Pressure Processing are the most studied methodologies. HTST is already utilized in some HMBs for daily practical activity and for research purposes. They seem to be superior to HoP for a better preservation of some nutritional and biologically protective components. Freeze-drying or lyophilization may have advantages for room temperature storage and transportation. The aim of this study is to evaluate the advancement regarding the processing of DHM with a literature search from 2019 to 2022. The effects of the new technologies on safety and quality of human milk are presented and discussed. The new technologies should assure microbiological safety of the final product at least at the same level as optimized HoP, with an improved preservation of the nutritional and bioactive components of raw human milk.

The indoor air quality should be better controlled and improved to avoid numerous health issues. Even if different devices are developed for air filtration, the proliferation of microorganisms under certain conditions must be controlled. For this purpose, a silver nanocluster/silica composite coating was deposited via a cosputtering technique onto fiber glass and polymeric based substrates. The aim of this work is focused on the evaluation of the antibacterial and antiviral effects of the developed coating. The preliminary results of the compositional and morphological tests showed an evenly distributed coating on filters surfaces. Several antibacterial tests were performed, confirming strong effect both in qualitative and quantitative methods, against S. epidermidis and E. coli. To understand if the coating can stop the proliferation of bacteria colonies spread on it, simulation of everyday usage of filters was performed, nebulizing bacteria solution with high colonies concentration and evaluating the inhibition of bacteria growth. Additionally, a deep understanding of the virucidal action and mechanism of Ag nanoclusters of the coating was performed. The effect of the coating both in aqueous medium and in dry methods was evaluated, in comparison with analysis on ions release. The virucidal performances are assessed against the human coronavirus OC43 strain (HCoV-OC43).

Despite the increasing body of evidence supporting the use of simulation in medicine, a question remains: when should we introduce it into the medical school's curriculum? We present the experience and future perspectives of the MD program in Medicine and Surgery of University of Turin-MedInTo. Since its launch, MedInTo has been dedicated to integrating innovative teaching approaches at the early stages into the medical curriculum. Herewith, we describe a case-based approach for our activities, which includes the utilization of simulation for emergency medical care training for students and the integration of virtual and augmented reality technology. Dedicated surgical training activities using virtual-augmented reality and life-like simulator for students are also described.