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Combination therapy targeting HDAC8 and SIRT1 improves muscle integrity in a dmd zebrafish model
No full textDuchenne Muscular Dystrophy (DMD) is a severe genetic disease that causes progressive muscle degeneration, inflammation, and fibrosis. Gene and cell therapies now offer promising therapeutic avenues, although their complexity makes pharmacological approaches complementary and more accessible. Histone deacetylases (HDACs) are a family of epigenetic regulators found deregulated in DMD patients and their modulation has emerged as a key strategy in DMD with Givinostat, a pan-HDAC inhibitor, recently approved by the FDA. However, the chronic use of pan inhibitors might be associated with side effects; while the treatment with selective compounds might reduce off-targets effects, maintaining the therapeutic ones. Our evidence demonstrated that the modulation of two HDACs , HDAC8 and SIRT1, was able to restore muscle function, acting on the stabilization of the cytoskeleton, and muscle energy metabolism, improving mitochondrial functioning . In dmd zebrafish embryos, the co-administration of the HDAC8 inhibitor PCI-34051, and the SIRT1 activator SRT2104, restores muscle loss and reduces inflammation. Importantly, this novel combination therapy allowed for lower doses of each compound, maximizing therapeutic effects and minimizing potential side effects. Moreover, we performed the acetylation profile of HDAC8 inhibited and SIRT1 activated zebrafish embryos to identify the respective acetylation targets. The identification of new targets holds significant potential for advancing therapeutic strategies.
This approach not only provides opportunities to refine existing treatments but also opens the door to developing entirely new strategies that may synergize with current interventions. Ultimately, these efforts aim to broaden the arsenal of pharmacological tools available for the treatment of DMD and similar conditions, offering improved outcomes for affected individuals
Archimedes. New Studies in the History and Philosophy of Science and Technology, Walter Roy Laird, The Renaissance of Mechanics. Ancient Science in the Age of Humanism, Springer Cham (2024), vol. 68
No full textRiflessioni sulle interpretazioni creative del giudice amministrativo in materia processuale
No full textIl presente intervento analizza, a partire dalle riflessioni contenute nel volume di A. Averardi, Sindacato sul potere pubblico e attivismo giudiziale, ETS, 2025, le tendenze creative del giudice amministrativo nell'interpretare la norma processuale e il ruolo dell'adunanza plenaria
Diagnostic Concordance of Two- and Three-Gene SARS-CoV-2 Molecular Assays in Cameroon: Implications for Efficient Pandemic Response in Low- and Middle-Income Countries
No full textBackground: The scale-up of molecular assays for diagnosing emerging pathogens has increased in low-and-middle-income countries (LMICs) since the advent of COVID-19. We herein evaluated the diagnostic concordance of three different assays for SARS-CoV-2 in Cameroon. Methods: A laboratory-based comparative study was performed on nasopharyngeal samples collected between March-2020 to March-2023 from the biobank of Chantal Biya International Reference Centre (CIRCB), Yaoundé-Cameroon. Samples were analyzed using DaAn Gene (N/ORF1ab-genes), ThermoFisher (N/ORF1ab/S-genes), and GeneXpert (N2/E-genes). Validated cycle thresholds (CT) for positivity were CT < 37 for DaAn Gene/ThermoFisher and CT < 40 for GeneXpert. Cohen's Kappa coefficient evaluated diagnostic concordance with DaAn Gene as reference. Results: We analysed 249 samples (55.8% males, median-age [IQR], 36 [27–50] years including 21.3% symptomatic participants). Overall positivity rates (median [IQR]) were 55.0% (CT: 30.6 [23.1–35.5]); 53.4% (CT: 26.6 [21.2–30.9]); 22.1% (CT: 32.7 [26.9–36.1]) for GeneXpert, DaAn Gene and ThermoFisher respectively. GeneXpert showed stronger concordance with DaAn Gene (83.1%; k = 0.66, 95% CI: 0.57–0.75) than ThermoFisher (67.9%; k = 0.38, 95% CI: 0.29–0.47). At validated thresholds, GeneXpert showed higher positive agreement with DaAn Gene (85.0%, 113/133) as compared to ThermoFisher (41.3%, 55/133), while maintaining comparable negative agreement (81.0% [GeneXpert] and 98.3% [ThermoFisher]). At low CTs (< 20) however, positive agreement with DaAn Gene was high for GeneXpert (100%, 15/15) and ThermoFisher (93.3%, 14/15). Conclusion: GeneXpert exhibits superiority over ThermoFisher in detecting cases of COVID-19. As expected, agreement between two- and three-genes assays at CT < 20 was excellent, suggesting interoperability of these platforms during outbreaks for high viral loads cases. However, two-genes assays may be decisive to guide decision-making for effective public health response while facing intermediate to low-level viral loads in LMICs
The Potential Role of Therapeutic Drug Monitoring for Safe and Effective Anti-Infective Therapy with Manipulated Dosage Forms
No full textBackground: Therapeutic drug monitoring (TDM) is essential for ensuring safe, effective, and individualized anti-infective therapy, particularly in patients with complex clinical needs. Variability in pharmacokinetics, challenges in drug administration, and high-dose regimens can compromise adherence and increase the risk of therapeutic failure or resistance. Swallowing difficulties, a common barrier to oral therapy, often necessitate alternative administration routes or customized formulations. However, interventions such as pharmaceutical compounding or manipulation of solid dosage forms may significantly alter drug bioavailability and pharmacokinetic profiles, making TDM indispensable for guiding dose adjustments and maintaining therapeutic targets. Objectives: This review not only emphasizes the clinical relevance of TDM but also addresses practical strategies that enable therapy when standard formulations are unsuitable or unavailable, while minimizing risks that could compromise treatment efficacy and safety. Special focus is given to anti-infective agents, such as antibiotics, antivirals, and antifungals, illustrating how TDM, combined with tailored pharmaceutical approaches, supports precision dosing and informed decision-making. Conclusions: Through clinical examples and pharmacokinetic considerations, we demonstrated that TDM is a cornerstone of personalized medicine, improving outcomes, and reducing adverse effects in anti-infective treatment
IS B4GALT5 DEFICIENCY A NEW CONGENITAL DISORDER OF GLYCOSYLATION?
No full textGlycosphingolipids (GSLs) are relevant components of mammalian cell membranes where they play multiple roles. Many specific glycosyltransferase enzymes are responsible for the synthesis of the different oligosaccharide chains of GSLs. Knock-out mouse models lacking single or multiple glycosyltransferases were generated in the past elucidating the role of individual GSLs in mouse brain. Inactive variants of two specific glycosyltransferases were found responsible for two human congenital disorders of glycosylation (CDG) characterized by clinical pictures not really predicted by the corresponding mouse KO models. Previous studies showed that two similar galactosyltransferases, B4GALT5 and B4GALT6, are both responsible for the synthesis of lactosylceramide (LacCer), the precursor of many GSLs. B4GALT5 appeared more relevant, but B4GALT6 was still able to rescue B4GALT5 KO in mice.
Genome analysis of a patient revealed a compound heterozygous mutation in B4GALT5 gene (DECIPHER Patient ID: 412221) opening the question if a novel human CDG exists not predicted by mouse models.
The aim of this thesis is to investigate if the above B4GALT5 variants are actually inactive and if human B4GALT6 is unable to compensate for the lack of B4GALT5 activity, indicating the possibility of such a novel CDG.
Cell models were constructed trough KO of either B4GALT5 or B4GALT6, or both together in HEK-293T cells using CRISPR/Cas9 genome editing. Transfections were performed with either coding cDNAs placed in expression vectors without or with N-terminal (Halo) or C-terminal (C-myc) tagging. Transcript levels by qPCR, protein expression by western blotting, galactosyltransferase activity by in vitro assay, and cell GSL biosynthesis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) were then determined. Similar analyses were also performed on patient’s serum and dermal fibroblasts.
Results showed that the mutations do not impair transcript levels or protein expression, while both variants are totally unable to use their substrates in vitro and to restore LacCer and GM3 ganglioside expression upon transfection in double KO HEK-293T cells. In silico studies of mutated proteins revealed pronounced conformational instability compared to the wildtype, affecting both protein backbone and UDP-Gal binding. ΔStability calculations confirmed strong destabilizing effects, particularly for G274R.
Moreover, the GSL profile of patient serum and fibroblasts displayed an accumulation of GlcCer and a strong reduction of LacCer, GM3 and globoside Gb3. These findings together indicate that both variants have lost their functional activity.
While B4GALT6 KO clones maintain almost unchanged the GSL pattern of HEK-293T cells, B4GALT5 KO clones retain less than 20% of the above GSLs, despite the transcript level of B4GLT6 is about 10-fold higher than that of B4GALT5. Moreover, upon transfection of B4GALT6 cDNA untagged or tagged at the C-terminus, no protein is detectable by western blotting, despite the high levels of B4GALT6 transcript achieved. Protein expression becomes evident only upon Halo tagging at the N-terminus, allowing detection of LacCer synthase activity in vitro and GSL rescuing in dKO clones. In particular, B4GALT6 ability to restore GSL expression is comparable to that of B4GALT5. However, detection of Halo-tagged B4GALT6 by western blotting is not comparable with that of Halo-tagged B4GALT5, since it requires much higher transcript levels to achieve comparable expression levels, as confirmed by transfecting the cDNAs placed under the control of weaker promoters. Hypothesizing a reduced efficiency of B4GALT6 translation, we constructed and transfected B4GALT6 chimeras where B4GALT5 N-terminal tail and transmembrane domain, or those plus the entire stem region, replaced the corresponding sequence of the native cDNA. Western blot analysis reveals detectable B4GALT6 protein with both chimeras, confirming the hypothesis.
In conclusion, the evidence that: (1) B4GALT5 variants reported in a patient are totally inactive; (2) human B4GALT6 is not efficiently translated being able to drive relevant GSL biosynthesis only at extremely high levels of its transcripts, suggest that the amounts of brain GSLs, probably gangliosides, could not be sufficient, at least in some cell types, to assure proper brain development and function. It is possible that a novel CDG exists, although additional patients and/or studied on differentiated neural cells or organoids are necessary for definitive proof
L'Affascinante spiritualità dell'Antico Egitto : Uno scavo archeologico svela i costumi della popolazione cosmopolita di Assuan
No full textLa presente pubblicazione accompagna la mostra multimediale "L'affascinante spiritualità dell'Antico Egitto. Uno scavo svela i costumi della popolazione cosmopolita di Assuan" ed espone i primi risultati dello scavo di EIMAWA (Egyptian-Italian Mission at West Aswan), diretto dall'Università degli Studi di Milano e dal Ministero del Turismo e delle Antichità egiziano
Education Research: Bridging the Artificial Intelligence Training Gap
No full textBackground and objectives: As artificial intelligence (AI) rapidly becomes an integral tool in clinical neurology, future clinicians will need to master its application in patient care. While previous studies focused primarily on medical students' perspectives, our survey, addressed to Italian neurology residents, aims to assess their familiarity with AI tools and identify educational needs of learners close to clinical practice and care delivery. Methods: A cross-sectional, web-based survey designed by the University of Milan was distributed nationwide to neurology residents from May 22 through July 30, 2025. The questionnaire included items on demographics, self-assessed AI knowledge, exposure to AI training, clinical applications, perceived challenges, and attitudes toward the impact of AI on neurology practice. Descriptive statistics and association analyses were performed. Results: A total of 173 residents (12.7%) completed the survey, with 37% affiliated with research hospitals. Although AI is frequently used in practice, with 40.7% using AI tools at least on a weekly basis, only 30.7% rated their knowledge as "good" or higher. Paradoxically, most residents (79.2%) reported no curriculum-integrated AI education, which was instead significantly associated with geographical background (Χ2 = 45.36, p < 0.001), while nearly all respondents expressed interest in further AI training (93.1%). The most familiar application was generative AI for clinical decision support (64.2%), and neuroimaging was rated as the first AI integration area (mean priority 2.99). Main barriers to AI application included concerns about reliability (71.1%) and data privacy (43.9%). Regarding career perspectives, 41.6% of residents believed that AI will create new job opportunities, whereas most (89%) agreed that the AI revolution will not replace human professionals. Discussion: Despite the limited number of participants, our survey provides a representative snapshot of AI knowledge, use, and attitudes among neurology residents in Italy, a critical country in the digital health care transformation, contributing important evidence for international shared recommendations on AI educational needs to support future clinical practice. Although AI tools are widely used, most residents had only basic knowledge and limited formal training, prompting calls for more structured, learner-centered educational modules to be integrated into neurology curricula
Association between early arterial pH, base excess and lactate and 24-h mortality and neurological outcomes after cardiac arrest and cardiopulmonary resuscitation: a translational study
No full textAim We aimed to assess the translational relevance of blood gas-derived acid-base parameters measured in rat and pig models of cardiac arrest and cardiopulmonary resuscitation, evaluating their potential as predictors of mortality and poor neurological outcome. Methods Seventy-seven rats, 83 pigs and 61 patients who experienced cardiac arrest of proven or suspected cardiac origin were retrospectively analyzed . Blood gas analyses were performed 4 h after return of spontaneous circulation. Neurological recovery was assessed using Neurological Deficit Score in rats, overall performance category in pigs, and cerebral performance category in patients. Nonlinear associations between blood gas-derived acid-base parameters and outcomes were analyzed using a generalized additive model. Receiver operating characteristics curve analyses were performed. Results In a multivariate regression analysis area under the curve, considering pH, base excess and lactate, for prediction of mortality were respectively: 0.796 (95%CI: 0.635–0.956), 0.980 (95%CI: 0.946–1.000), 0.959 (95%CI: 0.896–1.000) in rats; 0.908 (95%CI: 0.826–0.990), 0.933 (95%CI: 0.863–1.000), 0.798 (95%CI: 0.588–1.000) in pigs; and 0.830 (95%CI: 0.724–0.936), 0.832 (95%CI: 0.731–0.933), 0.839 (95%CI: 0.738–0.940) in patients. Area under the curve, considering pH, base excess and lactate, for prediction of poor neurological outcome were respectively: 0.673 (95%CI: 0.515–0.831), 0.724 (95%CI: 0.576–0.872), 0.900 (95%CI: 0.760–1.000) in pigs; and 0.835 (95%CI: 0.734–0.937), 0.835 (95%CI: 0.735–0.936), 0.884 (95%CI: 0.793–0.945) in patients. Conclusion Arterial pH, base excess and lactate were early independent predictors of both 24-h mortality and neurological outcome following cardiac arrest in animal models and in humans. BE showed the highest predictive value for mortality, while lactate was the strongest predictor for poor neurological outcome
