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Detection of equine herpesvirus type 1 (EHV-1) by recombinase polymerase amplification
No full textEquine herpesvirus type 1 is a major pathogen of horses, which must be detected quickly and effectively to isolate infected animals and prevent transmission to healthy horses. Laboratory diagnosis depends on a combination of serology and molecular biology, making it challenging to
establish a definitive field test. The aim of this study was to use recombinase technology for the field detection of this virus, thereby enabling a point-of-care test. Glycoprotein E (gE), one of the primary targets for detecting animal herpesviruses, was chosen for isothermal amplification with
a specific recombinase kit. Several primer, time, and temperature combinations were tested. The optimal amplification conditions were 41°C for 25 minutes. This combination proved effective for amplifying the gE fragment (226 bp), even when the template DNA was diluted 100-fold. These
parameters were subsequently applied in the point-of-care assay, which used a lateral flow detection device and produced comparable results, with a limit of detection ranging from 5 × 102 to 101 viral copies. The point-of-care test was compared to a real-time PCR method reported in the literature for field samples with known positivity/negativity. There was perfect agreement with negative samples (34/34), and 18 of 26 positives were accurately diagnosed. The assay identified EHV-1 DNA in 11/12 highly positive samples (with a low Ct in real-time PCR) and 7/14 weakly positive samples (Ct more than 26). The overall diagnostic specificity was 100%, and the sensitivity was 69.2%, which increased to 91.7% when highly positive samples were used. This work represents the first application of this technology for EHV-1 detection. Although viral load affects accuracy, this device has the potential to be used in the field, as it requires no specialized equipment and yields results rapidly
A novel multimodal imaging approach for working diagnosis of acute myocardial infarction with non-obstructive coronary arteries: a promising diagnostic strategy
No full textBackground: Myocardial infarction with non-obstructive coronary arteries (MINOCA) demands prompt mechanistic clarification. Early integration of coronary CT angiography (CCTA) and cardiovascular magnetic resonance (CMR) can refine diagnosis during the acute phase. Methods: Twenty-one consecutive patients (41 ± 10 years; 71% men) presenting with troponin-positive chest pain and unobstructed coronaries underwent CCTA, delayed iodine-enhanced CT for late iodine enhancement (LIE), and CMR imaging within 14 days, with a mean interval of 5 days [interquartile range (IQR) 2–9] between both imaging modalities. CCTA assessed luminal stenosis and high-risk plaque; LIE mapped iodine retention; CMR evaluated myocardial edema and late gadolinium enhancement (LGE). Clinical, electrocardiographic, and laboratory data were collected. Results: Eight patients were classified as MINOCA and 13 as acute myocarditis. Chest pain was universal; dyspnea and syncope occurred in seven and two patients, respectively. Median peak high-sensitivity troponin-I was 1,569 ng/L (IQR 589–5 771). Biventricular systolic function was preserved (mean LVEF 58%; RVEF 55%). LGE appeared in 16 subjects: subendocardial in every MINOCA case and intramural or subepicardial in eight myocarditis cases. Myocardial edema was present in 15 patients. CCTA showed no atherosclerosis in 16 patients; five displayed non-obstructive lesions (<50% stenosis) with high-risk plaque confined to three MINOCA subjects. LIE confirmed iodine uptake matching the LGE pattern in all MINOCA patients and in six with myocarditis. Conclusions: An acute CCTA-CMR protocol may aid in distinguishing ischemic from non-ischemic myocardial injury in presumed MINOCA and unmasks occult high-risk plaques. This multimodal imaging approach reveals occult high-risk coronary plaques and enhances diagnostic accuracy, thereby supporting mechanism-targeted management strategies in patients presenting with troponin-positive chest pain
Le comunità energetiche rinnovabili come strumenti privatistici di contrasto alla povertà energetica: profili strutturali, relazionali e funzionali
No full textInhibition of Enhancer of Zeste Homolog 2 alleviates oxidative stress, inflammation and barrier damage of human corneal epithelial cells
No full textDry Eye Disease (DED) is a multifactorial condition characterized by tear film instability, ocular surface inflammation, epithelial barrier dysfunction, and oxidative stress (OS). Recent studies have identified Enhancer of Zeste Homolog 2 (EZH2) as an important contributor to ocular surface diseases by promoting inflammation and epithelial dysfunction. Based on these findings, this study investigated the potential therapeutic effects of GSK343, a selective EZH2 inhibitor, in mitigating DED-associated pathological processes using in vitro models. Our findings demonstrated that low concentrations of GSK343 (1, 3 and 5 μM) did not compromise human corneal epithelial (HCE) cell viability and enhanced wound healing, suggesting its role in promoting epithelial regeneration. In H2O2-induced DED model, GSK343 pretreatment significantly restored HCE cell viability following H2O2-induced damage, reduced reactive oxygen species (ROS) levels and upregulated the expression of antioxidant enzymes as HO-1 and MnSOD, indicating a protective effect against OS. Additionally, GSK343 restored tight junction proteins (ZO-1 and occludin), reducing permeability and maintaining barrier integrity. In TNFα-induced DED model, GSK343 restored HCE cell viability and decreased the levels of pro-inflammatory cytokines IL-1β, TNFα, IL-6, and IL-17. Furthermore, GSK343 decreased the number of apoptotic cells and p53 positive cells, highlighting its protective properties. These findings suggest that GSK343 exerts anti-inflammatory, antioxidant, and epithelial-protective effects in DED models, suggesting EZH2 inhibition as a potential therapeutic strategy for DED management
Analisi biomeccanica delle espansioni mascellari simmetriche e asimmetriche con MARPE: uno studio tridimensionale agli elementi finiti (FEM)
No full textLa miniscrew-assisted rapid palatal expansion (MARPE) rappresenta una valida alternativa non chirurgica per la correzione della discrepanza trasversale del mascellare in pazienti tardo-adolescenti e adulti. Tuttavia, la biomeccanica dell’espansione, in particolare nei casi asimmetrici, non è ancora completamente prevedibile.
Questa tesi analizza, mediante uno studio tridimensionale agli elementi finiti (FEM), gli effetti di diverse configurazioni simmetriche e asimmetriche di MARPE, valutando l’influenza del numero e della posizione delle miniviti (TADs) sui pattern di spostamento scheletrico e sulla distribuzione delle tensioni nelle suture circum-mascellari. Il modello FEM è stato sviluppato a partire da una CBCT di un giovane adulto e sottoposto a un protocollo di attivazione clinicamente realistico.
I risultati mostrano che il posizionamento posteriore dei TADs incrementa l’espansione scheletrica, mentre configurazioni asimmetriche consentono espansioni differenziali controllate. I dati ottenuti supportano una pianificazione personalizzata della MARPE, migliorandone la prevedibilità biomeccanica e clinica
Italian clinical practice GRADE-based guidelines on the diagnosis and treatment of overweight and obesity, endorsed by the Italian National Institute of Health
No full textObesity is a chronic relapsing disease associated with increased morbidity and mortality and reduced quality of life. The present GRADE-based guidelines have been commissioned by the Italian Society of Obesity (SIO) by the Italian National Health Institute to provide evidence-based recommendations on obesity diagnosis and treatment. The panel identified 13 clinical questions, organised into four domains: A. diagnostic criteria (4 questions); B. medical nutrition therapy (4 questions); C. pharmacological, surgical, and endoscopic treatments (4 questions); and D. miscellaneous (1 question). The expert panel recommends adopting at least one anthropometric index of abdominal visceral fat distribution beyond body mass index (BMI) for better clinical risk stratification. A structured lifestyle intervention (i.e., medical–nutritional treatment—MNT), preferably based on cognitive–behavioural therapeutic approach and including physical activity and a balanced diet, should be offered to all subjects living with obesity. Pharmacological or surgical treatment should be offered in addition to MNT when MNT is unable to reach clinical goals. In particular, pharmacological treatment should be considered as the preferred option for subjects with BMI > 27 kg/m2 with comorbidities, and for those with BMI 30–39.9 kg/m2, based on individual therapeutic goals and needs, adopting surgical treatment in selected cases or in case of OMM failure to reach clinical goals. For subjects with a BMI > 40 kg/m2, surgical treatment may be considered conversely as a preferable option. Reduction of at least 10% of the initial body weight is associated with a better quality of life and a reduced risk of incident depression. The current guideline is endorsed by the Italian National Institute of Health, providing the new, updated clinical and legal reference for professionals involved in the management of subjects living with obesity in Italy
OTTIMIZZAZIONE DELLA SPECTRAL TC NELLA DIAGNOSI DEL CARCINOMA COLORETTALE: CONFRONTO TRA VMI E TC CONVENZIONALE; CORRELAZIONE SPETTRALE CON IL GRADING ISTOPATOLOGICO
No full textIntroduzione.
La TC convenzionale presenta una sensibilità limitata per la rilevazione del carcinoma del colon-retto (CRC). In questo contesto, la TC dual-energy (dlDECT) e le ricostruzioni spettrali potrebbero fornire biomarcatori quantitativi utili sia alla diagnosi sia alla stratificazione del rischio, integrando le informazioni morfologiche con parametri funzionali correlati a perfusione e composizione tissutale. Questo lavoro ha avuto tre obiettivi: (I) identificare il livello energetico ottimale delle VMI e valutarne il valore aggiunto diagnostico rispetto alla TC convenzionale; (II) correlare i parametri spettrali con il grading istopatologico (basso grado G1–G2 vs alto grado G3–G4); (III) individuare predittori indipendenti di alto grado e costruire un nomogramma di rischio.
Materiali e metodi.
Studio retrospettivo monocentrico condotto presso il Policlinico di Messina. Tra maggio 2021 e giugno 2025 sono stati valutati 245 pazienti con CRC istologicamente confermato sottoposti a dlDECT addominale per stadiazione, con intervallo ≤30 giorni tra TC e biopsia.
Per l’obiettivo (I) sono stati inclusi 108 soggetti (66 CRC e 42 controlli con endoscopia disponibile): sono state ricostruite VMI in fase arteriosa da 40 a 100 keV e calcolati SNR e CNR. La performance diagnostica è stata confrontata tra TC convenzionale e VMI al keV ottimale con analisi ROC.
Per gli obiettivi (II)–(III) è stata analizzata una coorte di 111 pazienti con CRC: sono state misurate in arteriosa e portale le HU su VMI, la concentrazione di iodio (IC) su mappe iodiniche, la concentrazione di iodio normalizzata (NIC) e la pendenza spettrale (λHU). Le associazioni con alto grado sono state valutate con analisi univariata e regressione logistica multivariata (backward) per identificare predittori indipendenti; è stato sviluppato un nomogramma e valutati discriminazione (AUC), calibrazione e decision curve analysis.
Risultati.
Obiettivo (I): La VMI 40 keV ha mostrato la massima attenuazione ed i migliori indici di qualità (SNR e CNR), significativamente superiori rispetto alla TC convenzionale ed alle altre energie. L’integrazione delle VMI a 40 keV ha migliorato in modo significativo la diagnosi di CRC: AUC da 0,875 a 0,943 nel lettore meno esperto e da 0,916 a 0,954 nel lettore più esperto, con incremento della sensibilità fino al 97%.
Obiettivo (II): I tumori di alto grado hanno mostrato valori più elevati di HU su VMI 40 in fase portale, IC in fase arteriosa e portale e λHU in fase portale; tra le variabili clinico-anatomiche, sesso e sede tumorale risultavano associati più frequentemente all’alto grado.
Obiettivo (III): al modello multivariato, la sede tumorale (OR 0,195) e la λHU in fase portale (OR 2,220) sono emerse come predittori indipendenti di alto grado. Il nomogramma derivato ha mostrato discriminazione soddisfacente (AUC 0,72), buona calibrazione (intercetta ~0; pendenza 1,0) e utilità clinica alla DCA con net benefit rispetto alle strategie “treat-all” e “treat-none” per soglie di probabilità circa 0,20–0,50.
Conclusioni.
La dlDECT migliora la rilevazione del CRC rispetto alla TC convenzionale. I parametri quantitativi spettrali risultano associati al grading istopatologico e, combinati con la sede tumorale, consentono un modello predittivo con potenziale applicazione preoperatoria per stratificazione del rischio e supporto decisionale
Uncertainty and biases on the determination of stability constants of metal complexes derived from potentiometric data. What can be expected?
No full textThe definition of reliable equilibrium constants is an essential step in speciation studies, as the uncertainty and the consistency of the estimated values concur to define the reliability of the speciation model. It is therefore necessary to evaluate the uncertainty contribution of all factors involved at each step of the data acquisition procedure and to be aware about the consequences of systematic errors on the best-estimated values of experimentally measured equilibrium constants. In this work, a series of computer-generated H+-ion selective electrode titration curves simulating the alkalimetric titration of equimolar mixtures of Zn2+-EDTA in aqueous solutions have been processed by different equilibrium data fitting software. Hence, the uncertainties of the refined stability constants of the Zn2+-EDTA complexes could be derived, while excluding the experimental variability that intrinsically affects all practical experiments. The sensitivity of a given chemical system to different data processing strategies and to possible errors in the input data was evaluated. The systematic errors considered relate to the potential reading, the concentration of the titrant and solution components, and ionic strength variations during titrations. The outcomes highlight how unsuitable decisions taken at the stage of nonlinear least squares fitting of the data can affect the results and underline that the main error contribution is related to the measurement of the glass-electrode potential. The processing of simulated data sets can be a useful tool to alert the researchers to the sensitivity of a given chemical system to different strategies and to possible errors in the input data
From the Memorial of Deir ez-Zor to the Cathedral of Shushi: Cultural Cleansing and Transnationalism
No full textThis chapter focuses on heritage memory as representative of a culture of belonging. Focusing on Syrian and Karabakh Wars, the chapter aims to shed light on the determinants of the contemporary conflicts in the area; particularly those that arise when heritage memory is denied and the process of “cultural cleansing” acquires transnational dimensions. The authors ask, how does the destruction of cultural heritage erase the collective memory of a people
