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3D-tumor sphere (SPH) cultures enriched in CSCs were generated from intrahepatic CCA (iCCA) cell lines and CCA patient-derived organoids (PDOs) were employed. Circulating monocytes were differentiated into M2c MØs and recombinant Gas-6, a MerTK ligand, was used to activate MerTK. MERTK expression in human CCA tissues was analyzed at mRNA level in public database (n=78) and confirmed by immunohistochemistry (IMH) (n=74). Single-cell RNA sequencing of CD45+ sorted cells was performed in paired non-tumoral and tumoral specimens from intrahepatic CCA patients (n=6). Conditioned media (CM) of iCCA SPH induced higher MerTK expression in macrophages. Conversely, soluble mediators released by Gas-6-stimulated M2c MØs, which express MerTK at high levels, increased sphere number and volume, expression of stem-like genes and drug-resistance in iCCA cells. Moreover, the exposure to CM of M2c MØs induced an increase in the cell viability of organoids CCA PDOs, which was further increased when macrophages were stimulated with Gas-6. These effects were reduced following treatment of macrophages with UNC2025, a small molecule inhibitor of MerTK. Transcriptomic analysis of laser-captured, micro-dissected epithelium and stroma from 23 iCCA patients showed that MerTK mRNA expression is significantly higher in intratumoral stroma. These data were further confirmed in a public iCCA dataset showing that high MerTK levels are associated with immunologically hot iCCA. Single-cell RNA sequencing of CD45+ cells from non-tumoral and tumoral areas in iCCA patients showed that MerTK is predominantly expressed at the level of myeloid cells. Further reclustering showed MerTK expression in ID3 MØs, corresponding to Kupffer cells in tumor tissue. Notably, expression of MerTK correlated with greater tumor size, tumor grade, microvascular invasion, and risk of recurrence in iCCA patients. These data indicate that a cross-talk between MerTK-expressing cells in the stroma and iCCA cells results in increased malignant features. [ABSTRACT FROM AUTHOR]

PSVD includes a group of vascular diseases featuring lesions encompassing portal venules and sinusoids, irrespective of the presence of portal hypertension (PH). Liver biopsy (LB) is fundamental for diagnosis. A recent monocenter study showed high rates of PSVD in patients with isolated and unexplained GGT elevation who underwent a LB; due to the small sample, the study could not identify factors associated with PSVD. Aim of this study was to validate in a multicenter cohort the rate of PSVD in patients with isolated GGT elevation and identify clinical factors associated with PSVD. We retrospectively included all patients who underwent a LB for unexplained isolated and persistent GGT elevation in five Italian Hepatology Units from March, 2015 to December, 2021. All LB specimens were reviewed by hepatopathologists. 144 patients met the inclusion criteria and were enrolled in the study. Most patients were males (76/144, 53%) with the mean age being 52 years (range 19-74). 11 patients (7.6%) presented liver stiffness (LS) suggestive of advanced liver disease (>10 kPa) but none showed clinical features of PH. The histological findings were compatible with PSVD in 96 out of 144 patients (67%). In 13 patients (9%) histology was compatible with NASH, in 3 patients (2%) with hepatic sarcoidosis and in 3 patients with congenital hepatic fibrosis (CHF). Histology showed healthy liver in 29 (20%) patients. At univariate analyses male sex, LS <10 kPa and GGT <214 UI/L were associated with PSVD (p<0.05). The same factors were confirmed by logistic regression: LS <10 kPa (OR 7.5 (95%, CI 1.7-32.3), GGT <214 UI/L (OR 2.8 (95%, CI 1.29-6.28) and male gender (OR 2.5 (95%, CI 1.19-5.58). PSVD is common in patients with elevated GGT and no signs of PH. Male gender, LS and GGT levels can be used to identify these patients a priori. [ABSTRACT FROM AUTHOR]

BackgroundTransition to digital pathology usually takes months or years to be completed. We were familiarizing ourselves with digital pathology solutions at the time when the COVID-19 outbreak forced us to embark on an abrupt transition to digital pathology. ObjectiveThe aim of this study was to quantitatively describe how the abrupt transition to digital pathology might affect the quality of diagnoses, model possible causes by probabilistic modeling, and qualitatively gauge the perception of this abrupt transition. MethodsA total of 17 pathologists and residents participated in this study; these participants reviewed 25 additional test cases from the archives and completed a final psychologic survey. For each case, participants performed several different diagnostic tasks, and their results were recorded and compared with the original diagnoses performed using the gold standard method (ie, conventional microscopy). We performed Bayesian data analysis with probabilistic modeling. ResultsThe overall analysis, comprising 1345 different items, resulted in a 9% (117/1345) error rate in using digital slides. The task of differentiating a neoplastic process from a nonneoplastic one accounted for an error rate of 10.7% (42/392), whereas the distinction of a malignant process from a benign one accounted for an error rate of 4.2% (11/258). Apart from residents, senior pathologists generated most discrepancies (7.9%, 13/164). Our model showed that these differences among career levels persisted even after adjusting for other factors. ConclusionsOur findings are in line with previous findings, emphasizing that the duration of transition (ie, lengthy or abrupt) might not influence the diagnostic performance. Moreover, our findings highlight that senior pathologists may be limited by a digital gap, which may negatively affect their performance with digital pathology. These results can guide the process of digital transition in the field of pathology.

Hepatitis B virus (HBV) infection represents a serious global health problem and persistent HBV infection is associated with an increased risk of cirrhosis, hepatocellular carcinoma and liver failure. Recently, the study of the role of microRNA (miRNA) in the pathogenesis of HBV has gained considerable interest as well as new treatments against this pathogen have been approved. A few studies have investigated the antiviral activity of vitamin E (VE) in chronic HBV carriers. Herein, we review the possible role of tocopherols in the modulation of host miRNA with potential anti-HBV activity. A systematic research of the scientific literature was performed by searching the MEDLINE, Cochrane Library and EMBASE databases. The keywords used were ‘HBV therapy’, ‘HBV treatment’, ‘VE antiviral effects’, ‘tocopherol antiviral activity’, ‘miRNA antiviral activity’ and ‘VE microRNA’. Reports describing the role of miRNA in the regulation of HBV life cycle, in vitro and in vivo available studies reporting the effects of VE on miRNA expression profiles and epigenetic networks, and clinical trials reporting the use of VE in patients with HBV-related chronic hepatitis were identified and examined. Based on the clinical results obtained in VE-treated chronic HBV carriers, we provide a reliable hypothesis for the possible role of this vitamin in the modulation of host miRNA profiles perturbed by this viral pathogen and in the regulation of some cellular miRNA with a suggested potential anti-HBV activity. This approach may contribute to the improvement of our understanding of pathogenetic mechanisms involved in HBV infection and increase the possibility of its management and treatment. [ABSTRACT FROM PUBLISHER]

Mast cells are essential not only for allergies but also for innate and acquired immunity, autoimmunity and inflammation, and they are recognized as a new type of immunoregulatory cells capable of producing different cytokines. Natural compounds have long been recognized to possess anti-inflammatory, antioxidant and anticancergenic activity. Quercitin is an inhibitor for mast cells and is a potent antioxidant, cytoprotective and anti-inflammatory compound and has a negative effect on intracellular regulator signal events initiated by FceRI receptor cross-linking and other activating receptors on mast cells. These observations candidate quercitin as a therapeutic compound in association with other therapeutic molecules. Capsaicin is a compound derived from peppers, especially capsicum, and is involved in stimulating circulation aiding digestion and relieving pain. Capsaicin receptor sub type I (VRI) is expressing in neurons and is present in a number of brain nuclei and in non-neuronal tissues, mediating inflammatory response. Capsaicin is involved in migraine, allergic symptoms, arthritis pain and gastric secretion. In this paper we review the biological effects of quercitin and capsaicin. [ABSTRACT FROM AUTHOR]