Kaposi sarcoma herpesvirus/human herpesvirus-8 (HHV-8) neoplastic and non-neoplastic disease in solid organ transplant (SOT) recipients can be life-threatening. We evaluated the seroprevalence of HHV-8 infection among donors (D) and recipients (R), the incidence of HHV-8 transmission/reactivation, and the clinical characteristics, management, and outcomes of HHV-8-related diseases, including Kaposi sarcoma herpesvirus-associated inflammatory cytokine syndrome (KICS), in consecutive SOT patients from 2011 to 2023. HHV-8 seroprevalence was 3.3% in 1349 donors and 8.4% in 1856 recipients screened (p<0.0001). Among the D+/R- group (n=49), 13 patients developed HHV-8-related diseases: 7 liver recipients had KICS, and 1 lung recipient had Kaposi sarcoma (KS) with subsequent KICS. Four KICS patients treated with rituximab survived, while the 3 patients not treated with rituximab died. Within the D-/R- group, out of 5 (0.3%) patients with non-donor-derived primary HHV-8 infection, 3 liver recipients developed KICS. In the R+ patients (n=155), 3 developed KS. In our cohort, 25/944 (2.6%) liver transplant recipients had a primary HHV-8 infection, and 10 of them (40%) developed KICS; 40% (4/10) of HHV-8 seropositive heart transplant recipients developed reactivation and 2 of them (50%) had fatal KS. Serologic screening and molecular surveillance of D+/R- patient groups facilitate early recognition and effective therapy of KICS.

Mesenchymal stromal/stem cells (MSCs) have emerged as a therapeutic tool in regenerative medicine. Recent studies have shown that exosome (EXO)-derived microRNAs (miRNAs) play a crucial role in mediating MSC functions. Additionally, intracellular miRNAs have been found to regulate MSC therapeutic capacities. However, the molecular mechanisms underlying miRNA-mediated MSC effects are not fully understood. We used 3D culture and IFN-γ to prime/enhance the MSC therapeutic effects in terms of functional miRNAs. After priming, our analysis revealed stable variations in intracellular miRNA among the MSC biological replicates. Conversely, a significant variability of miRNA was observed among EXOs released from biological replicates of the priming treatment. For each priming, we observed distinct miRNA expression profiles between the MSCs and their EXOs. Moreover, in both types of priming, gene ontology (GO) analysis of deregulated miRNAs highlighted their involvement in tissue repair/regeneration pathways. In particular, the 3D culture enhanced angiogenic properties in both MSCs and EXOs, while IFN-γ treatment enriched miRNAs associated with immunomodulatory pathways. These findings suggest that 3D culture and IFN-γ treatment are promising strategies for enhancing the therapeutic potential of MSCs by modulating miRNA expression. Additionally, the identified miRNAs may contribute to understanding the molecular mechanisms underlying the miRNA-mediated therapeutic effects of MSCs.

Across recent decades the increasing prevalence of multi-drug resistant KPC-carrying K. pneumoniae (KPC-Kp) has become a worldwide public concern. Herein, we characterized a ceftazidime/avibactam (CAZ/AVI), meropenem/vaborbactam (MER-VAB) and imipenem/relebactam (IMI-REL) resistant KPC-Kp strain isolated from a critically ill transplanted patient. Antimicrobial susceptibility test and whole-genome sequencing (WGS) were conducted to characterize the strain at the phenotypic and the genotypic levels. Genomic DNA was sequenced using the Illumina platform. Bioinformatic analyses were used to investigate the genome sequences both for resistance and virulence features, and for characterization of plasmids. The phenotypic characterization revealed that the KPC-Kp isolate was highly resistant to a wide range of antibiotics, including all the beta-lactam/beta lactamase inhibitor combinations, such as CAZ/AVI, MER-VAB, IMI-REL, and cefiderocol (FDC). WGS analysis showed that the isolate, belonging to the rare lineage ST661, contained several resistance and virulence genes. Among the resistance genes, we identified a new KPC variant, inside the mobile genetic element Tn4401, KPC-245, characterized by an insertion of 9 aminoacids (RAPNKDDYT) at position 263 and an aminoacidic change E274D of the protein sequence, compared to KPC-3. Interestingly, the presence of mutations only in bla_KPC gene, and not in other beta-lactamases coding genes, strongly points KPC-245 role in beta-lactam/beta-lactamase inhibitor combinations and FDC resistance. In our study, by using WGS analysis on a clinical isolate, we identified a new bla_KPC variant inside the Tn4401 transposon. Our results confirm the important role of the continuous surveillance of MDR K. pneumoniae in the clinical context.

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder that presents with heterogeneous clinical and pathological features, necessitating improved biomarkers for accurate diagnosis and patient stratification. In this study, we applied a data-independent acquisition (DIA)-based proteomics workflow to cerebrospinal fluid (CSF) samples from 138 individuals, including AD patients with high (Aβ+/tau+) or normal (Aβ+/tau-) CSF tau levels, and non-AD controls. Analysis using an Astral mass spectrometer enabled unprecedented proteome depth, identifying 2661 proteins with high data completeness. Comparative proteomic profiling revealed distinct protein signatures for Aβ+/tau+ and Aβ+/tau- subtypes. These findings were validated using an independent internal cohort and further corroborated with publicly available datasets from larger external AD cohorts, demonstrating the robustness and reproducibility of our results. Using machine learning, we identified a panel of 15 protein classifiers that accurately distinguished the two AD subtypes and controls across datasets. Notably, several of these proteins were elevated in the preclinical stage, underscoring their potential utility for early diagnosis and stratification. Together, our results demonstrate the power of DIA proteomics on the Astral platform, combined with machine learning, to uncover subtype-specific biomarkers of AD and support the development of personalized diagnostic strategies.

Background/Objectives: Type 1 diabetes (T1D) is a chronic autoimmune condition characterized by the destruction of pancreatic β-cells, leading to insulin deficiency. Current therapies, such as islet transplantation, face significant challenges, including limited donor availability and the need for lifelong immunosuppression. Encapsulation technologies offer a promising alternative, providing immune protection and maintaining β-cell viability. In this study, we propose an encapsulation device featuring a spiral tubular semipermeable polyethersulfone (PES) membrane reinforced with a rigid biocompatible resin scaffold. Methods: The PES membrane was engineered with a tailored porosity of 0.5 µm, enabling efficient nutrient and oxygen exchange while preventing immune cell infiltration. Using INS-1E insulin-secreting cells aggregated into size-controlled islet-like spheroids (ILSs), we evaluated the device’s performance. Results: The device achieved high ILS viability and insulin secretion over 48 h at therapeutic densities, maintaining functionality comparable to free-floating ILSs (control). The PES membrane, with its mechanical stability and biocompatibility, ensured durability without compromising diffusion dynamics, overcoming a critical limitation of other encapsulation approaches. Importantly, the device geometry allowed for the encapsulation of up to 356,000 islet equivalents (IEQs) in a single capillary fiber, reaching therapeutic thresholds for T1D patients. Conclusions: this device, with its innovative design, enables high-density encapsulation while preserving ILS functionality and scalability, making it a potential platform for clinical application. This work highlights the potential of PES-based encapsulation devices to overcome key barriers in T1D treatment, paving the way for personalized, long-term solutions to restore insulin independence.

Advanced heart failure is characterized by profound autonomic dysfunction, yet resting measures of heart rate variability (HRV) provide limited insight into autonomic adaptability under physiological stress. We investigated whether graded exercise unmasks alterations in cardiac autonomic regulation in patients with advanced heart failure using both conventional and information-theoretic HRV measures. 77 patients with end-stage heart failure with reduced ejection fraction underwent resting and cycle-ergometer exercise testing with continuous electrocardiographic recording. HRV was assessed at rest, during early exercise and at peak exercise using time- and frequency-domain indices alongside entropy-based measures of cardiac rhythm complexity. Clinical follow-up was performed for a composite adverse outcome, defined as cardiovascular death, comprising cardiac-related death, heart transplantation or left ventricular assist device implantation. During graded exercise, conventional HRV measures showed the expected workload-dependent attenuation, whereas entropy-based metrics revealed a progressive increase in signal complexity and a reduction in heartbeat predictability. Notably, during the early phase of exercise, patients who subsequently experienced adverse outcomes (n = 17) exhibited lower conditional entropy and higher self-entropy compared with survivors, while no differences were observed at rest or peak exercise. In survival analyses, self-entropy measured during early exercise remained independently associated with adverse outcomes after adjustment for clinical and exercise-derived variables. These findings indicate that early exercise represents a sensitive physiological window for revealing impaired autonomic adaptability in advanced heart failure. Information-theoretic analysis of HRV during submaximal exercise may provide complementary insights into cardiac autonomic regulation that are not captured by conventional resting or peak exercise measures.