Metabolic reprogramming of tumour cells sustains cancer progression. Similar to other cancer cells, glioblastoma cells exhibit an increased glycolytic flow, which encourages the use of antiglycolytics as an effective complementary therapy. We used the antiglycolytic 3-bromopyruvate (3BP) as a metabolic modifier to treat U118 glioblastoma cells and investigated the toxic effects and the conditions to increase drug effectiveness at the lowest concentration. Cellular vitality was not affected by 3BP concentrations lower than 40 μM, although p-Akt dephosphorylation, p53 degradation, and ATP reduction occurred already at 30 μM 3BP. ROS generated in mitochondria were enhanced at 30 μM 3BP, possibly by unbalancing their generation and their disposal because of glutathione peroxidase inhibition. ROS triggered JNK and ERK phosphorylation, and cyt c release outside mitochondria, not accompanied by caspases-9 and -3 activation, probably due to 3BP-dependent alkylation of cysteine residues at caspase-9 catalytic site. To explore the possibility of sensitizing cells to 3BP treatment, we exploited 3BP effects on mitochondria by using 30 μM 3BP in association with antimycin A or menadione concentrations that in themselves exhibit poor toxicity. 3BP effect on cyt c release and cell vitality loss was potentiated due the greater oxidative stress induced by antimycin or menadione association with 3BP, supporting a preeminent role of mitochondrial ROS in 3BP toxicity. Indeed, the scavenger of mitochondrial superoxide MitoTEMPO counteracted 3BP-induced cyt c release and weakened the potentiating effect of 3BP/antimycin association. In conclusion, the biochemical mechanisms leading U118 glioblastoma cells to viability loss following 3BP treatment rely on mitochondrial ROS-dependent pathways. Their potentiation at low 3BP concentrations is consistent with the goal to minimize the toxic effect of the drug towards non-cancer cells.

Wastewater surveillance has proven to be a cost-effective, non-invasive method for monitoring the spread and evolution of SARS-CoV-2, yet its value during today’s low-incidence phase is still being defined. Between August 2023 and July 2024, 42 composite wastewater samples were collected in Perugia, Italy and analyzed using RT-qPCR and whole-genome sequencing to identify circulating SARS-CoV-2 lineages. In parallel, clinical samples (respiratory tract samples) were collected and analyzed, allowing for direct comparisons to confirm the robustness of the wastewater findings. The sewage viral loads ranged from 8.9 × 10⁵ to 4.9 × 10⁷ genome copies inhabitant⁻¹ day⁻¹, outlining two modest community waves (September–December 2023 and May–July 2024). Sequencing resolved 403 Omicron lineages and revealed three successive subvariant phases: (i) XBB.* dominance (August–October 2023), when late-Omicron XBB subvariants (mainly EG.5.* and XBB.1.5) accounted for almost all genomes; (ii) a BA.2.86/JN surge (November 2023–March 2024), during which the BA.2.86 subvariant, driven mainly by its JN descendants (especially JN.1), rapidly displaced XBB.* and peaked at 89% in February 2024; and (iii) KP.* takeover (April–July 2024), with JN.1-derived KP subvariants rising steadily and KP.3 reaching 81% by July 2024, thereby becoming the dominant lineage. Comparisons of data from wastewater and clinical surveillance demonstrated how the former presented a much higher diversity of circulating viral lineages. Importantly, some subvariants (including BA.2.86*) were detected in wastewater weeks to months prior to clinical identification, and for longer periods. Taken together, the obtained data validated wastewater surveillance as an effective early warning system, especially during periods of low infection prevalence and/or limited molecular testing efforts. This methodology can thus complement clinical surveillance by offering valuable insights into viral dynamics at the community level and enhancing pandemic preparedness.

Legionella pneumophila, a waterborne pathogen naturally present in freshwater and capable of colonizing artificial water systems, is responsible for Legionnaires’ disease (LD), a severe form of pneumonia transmitted through inhalation of contaminated aerosols. Virulence of Legionella strains is affected by the plasticity of their genome, shaped by horizontal gene transfer and recombination events. Thus, contaminated water systems can host diverse Legionella populations with a distinct virulence potential. Here, we compare the genomic diversity of Legionella pneumophila strains isolated in water systems of academic buildings, together with their cytotoxicity and intracellular replication in THP-1-like macrophages. A six-year environmental surveillance revealed Legionella pneumophila contamination in 20 out of the 50 monitored sites, identifying five serogroups (sg) and 13 Sequence Types (STs). Phylogenetic investigations based on core genome multilocus sequence typing (cgMLST) and comparative genomics of representative isolates of each ST showed a broad diversity and a heterogeneous virulence repertoire, especially within the Dot/Icm and Lvh secretion systems. Following macrophage infection, a strain-dependent cytotoxicity and intracellular replication was observed, underlying significant pathogenic diversity within the same species and stage-dependent infection dynamics. Together, these results showed strain-specific genetic and phenotypic virulence traits to be considered during risk assessment in environmental surveillance.

Antimicrobial resistance (AMR) is increasingly recognized as an environmental issue, with wastewater treatment plants (WWTPs) acting both as receptors and potential dispersal points. This study compared a conventional activated sludge (CAS) and a membrane bioreactor (MBR, hollow-fiber, 0.04 µm) operating in parallel at a full-scale municipal WWTP (120,000 population equivalent, Trento, Italy), fed by the same primary-settled wastewater. Over six months, 24-h composite samples were collected at four points (raw influent, primary-settled wastewater, CAS effluent, MBR permeate) and analyzed by qPCR for six targets (16S rRNA, intI-1, qnrS, blaCTX-M, blaKPC, vanA) and by shotgun metagenomics. Primary settling had negligible impact on ARG abundance (0.05 log removal), whereas biological treatment achieved substantial reductions, with the MBR (3.3 logs, up to 4.9) significantly outperforming CAS (1.9 logs). Removal was gene-specific, highest for qnrS and blaCTX-M, and correlated with influent flow rate, mixed-liquor suspended solids, and organic matter. Metagenomics showed that the MBR reduced not only abundance but also the richness and diversity of bacteria and resistance genes, limiting their potential to spread further. These findings support MBR technology as a robust, though not absolute, barrier against AMR dissemination, pointing to the role that quaternary treatment could play in future wastewater management.

Background/Objectives: Genetic variation in bitter taste receptors, particularly TAS2R38, influences sensitivity to bitter compounds such as 6-n-propylthiouracil (PROP). However, its influence on food preferences and dietary habits remains unclear. This study investigated the relationship between TAS2R38 rs713598 polymorphism, a variant in strong linkage disequilibrium with other common variants and therefore served as a proxy for TAS2R38 bitter taste-related genetic variation, PROP responsiveness, food preferences, and adherence to the Mediterranean diet. Methods: A cross-sectional molecular epidemiological study was conducted on 200 Italian adults. Participants completed questionnaires assessing food preferences, Mediterranean diet adherence (energy-restricted Mediterranean Diet Adherence Screener; er-MEDAS), and physical activity (IPAQ). Bitter taste perception was evaluated using the PROP test and a Labelled Magnitude Scale. Saliva samples were collected for TAS2R38 rs713598 genotyping by qPCR. Associations were analysed using Spearman’s correlation and generalised linear models. Results: Genotyping was completed for 176 participants. A significant positive correlation was found between rs713598 genotype and PROP sensitivity (ρ = 0.404, p < 0.01). Women showed higher PROP responsiveness than men. No significant associations were observed between PROP responsiveness or TAS2R38 genotype and either food preferences or Mediterranean diet adherence. Older age was associated with greater preference for bitter foods (p = 0.008), whereas higher physical activity and non-smoking status were associated with greater adherence to the Mediterranean diet (p = 0.020 and p = 0.024, respectively). Conclusions: TAS2R38 polymorphism influenced bitter taste perception but showed limited relevance for food preferences and dietary habits. Lifestyle factors appeared more influential in shaping dietary behaviour.