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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]

Abstract: Background: The WHO-classification was shown to be an independent prognostic marker in some but not all retrospective studies possibly due to lack of reproducibility. We investigated the reproducibility of the WHO-classification and its prognostic implication using a large series of resected thymomas. Methods: Four independent pathologists histologically classified a surgical series of 129 thymic tumors in a blinded fashion. Fleiss’ kappa-coefficient was used to assess the pathologists’ overall agreement, and Cohen-Kappa to assess the agreement between two observers. Disease-related-survival (DRS) and progression-free-survival (PFS) curves were generated by Kaplan–Meier method and compared by log-rank test. Results: In 63/129 (48.8%) cases there was a complete agreement; in 43/129 (33.3%) cases 3/4 pathological diagnoses were identical; in 15/129 (11.6%) cases the diagnoses were identical by pair; in 8/129 (6.2%) cases three different pathological diagnoses were on record. The Kappa-correlation coefficient was only moderate (0.53). A following web review carried out on the 23 cases with at least two different diagnoses reached a complete consensus. The histotype showed a statistically significant impact on PFS and DRS in the classification provided by only two pathologists. Conclusions: In this study, the agreement on WHO classification of thymomas was only moderate and this impacted on patients management. Web consensus conference on the diagnosis, more stringent diagnostic criteria or the adoption of referral diagnostic centres may substantially reduce discrepancies. [Copyright &y& Elsevier]

Abstract: T1N0M0 (stage I) breast cancer (BC) has been increasing in recent decades but the optimal adjuvant approach remains controversial. To assess the outcome of BC patients stratified and treated with multimodal therapies according to St. Gallen consensus meeting recommendations, we retrospectively evaluated an unselected cohort of T1N0M0 BC patients, with respect to the St. Gallen criteria. At a median follow-up of 5 years, the recurrence rate, recurrence-free survival and overall survival were 7%, 94% and 96% respectively, and 60% of relapses were locoregional. No statistically significant difference was observed between T1a,b/T1c groups, or among risk categories (high/intermediate/low). The very low rate of distant recurrences even in patients with unfavorable prognostic factors seems to support the use of adjuvant systemic therapies but better prognostic and predictive factors are strongly needed for this subset of patients. [Copyright &y& Elsevier]

The impact of KRAS mutations on cetuximab sensitivity in epidermal growth factor receptor fluorescence in situ hybridisation-positive (EGFR FISH+) metastatic colorectal cancer patients (mCRC) has not been previously investigated. In the present study, we analysed KRAS, BRAF, PI3KCA, MET, and IGF1R in 85 mCRC treated with cetuximab-based therapy in whom EGFR status was known. KRAS mutations (52.5%) negatively affected response only in EGFR FISH+ patients. EGFR FISH+/KRAS mutated had a significantly lower response rate (P=0.04) than EGFR FISH+/KRAS wild type patients. Four EGFR FISH+ patients with KRAS mutations responded to cetuximab therapy. BRAF was mutated in 5.0% of patients and none responded to the therapy. PI3KCA mutations (17.7%) were not associated to cetuximab sensitivity. Patients overexpressing IGF1R (74.3%) had significantly longer survival than patients with low IGF1R expression (P=0.006), with no difference in response rate. IGF1R gene amplification was not detected, and only two (2.6%) patients, both responders, had MET gene amplification. In conclusion, KRAS mutations are associated with cetuximab failure in EGFR FISH+ mCRC, even if it does not preclude response. The rarity of MET and IGF1R gene amplification suggests a marginal role in primary resistance. The potential prognostic implication of IGF1R expression merits further evaluation. [ABSTRACT FROM AUTHOR]

Objective To determine the effect on kidney oxalate-salt deposition of a prolonged diet which induced vitamin B6 deficiency in adult rats, as there are reports of the pathogenic involvement of vitamin B6 deficiency in the formation of renal calcium oxalate calculi. Materials and methods The study comprised 24 6-month-old male albino Wistar rats; 12 were fed with a purified vitamin B6 -deficient diet and the others provided with the same diet but supplemented with 6 mg/kg of vitamin B6 . After 12 weeks, all rats were killed, and their kidneys fixed in formalin and routinely processed to paraffin for morphological examination; some fragments were fixed in glutaraldehyde and prepared for ultrastructural examination. From each rat consecutive sections of both kidneys were cut and stained with haematoxylin and eosin, periodic-acid Schiff, Sirius red and the Von Kossa method for calcium. Sections were examined in polarized light and by electron microscopy. Results The histopathological and ultrastructural features of the kidney of vitamin B6 -deficient rats were those of tubular-interstitial nephritis, characterized by tubular atrophy, interstitial fibrosis and chronic inflammatory infiltration. Oxalate and phosphate crystals were present in the papillary and parenchymal connective tissue. Ultrastructural features confirmed severe tubular epithelial lesions and the presence of an interstitial and intraepithelial inflammatory infiltrate; there was mild interstitial fibrosis. None of these features were apparent in the kidney of control rats. Conclusions Histopathological and ultrastructural data indicate that a prolonged vitamin B6 -deficient diet may contribute to the formation and deposition of calcium phosphate and oxalate crystals, which lead to severe damage of the renal parenchyma. This phenomenon may occur not only in growing rats, which have more active protein metabolism and consequently higher... [ABSTRACT FROM AUTHOR]

Objective: To describe our experience of endovascular repair of paraanastomotic aortic aneurysm. Methods and results: From March 2001 to December 2004 we identified 6 patients with a paraanastomotic aortic aneurysms following previous open repair of abdominal aortic aneurysm. All patients were treated with endovascular surgery under epidural anaesthesia. There were no major complications, surgical conversions or deaths. Four patients received a bifurcated aortic stent-graft, and two an aorto-uniliac stent-graft followed by a femoro-femoral bypass. At follow-up (mean 26.1±10.2 months) there were no deaths, endoleaks or graft migrations observed. Conclusion: Endovascular surgery, avoiding general anesthesia and re-laparotomy, is the ideal technique for treatment of this complication resulting from failed primary conventional AAA repair. [Copyright &y& Elsevier]

The treatment of benign broncho-oesophageal fistula is usually surgical but sometimes other methods of treatment are preferred. The case history is described of an adult in poor general condition in which an endoscopic approach combined with the use of fibrin glue on the bronchial side and metallic clips on the oesophageal side was used to close the fistula. [ABSTRACT FROM AUTHOR]

Summary: Surgical drapes are a physical barrier between the surgical team''s sterile working space (the operating field) and the non-sterile environment of the anaesthesia team and the nursing team within the operating theatre. Once the patient''s position has been checked and the skin preparation has been done, they are put in place. They are usually opaque, leaving visible just the areas where the surgical and anaesthesia teams will be working. When the patient''s position is changed perioperatively, malpositions may be unnoticed and cause complications. Using a transparent head drape allows one to check the position of the head and upper limbs at all times, thus reducing the risk of complications. It is a simple, easy, economic, non-restrictive method which will produce an improvement in the quality of patient care. [Copyright &y& Elsevier]

We report the case of a 34-year-old woman affected by breast cancer that had metastasized to the bone. She had been treated with oral cyclophosphamide and methotrexate (metronomic chemotherapy) and achieved 3.5 years of clinical remission. To our knowledge, this is the first description of such a prolonged response to therapy. This case report adds weight to known data on metronomic treatment and supports further investigation of this therapy. [ABSTRACT FROM AUTHOR]

A computer-aided image analysis system was applied to evaluate the Sirius-red stained collagen fibers amount (CF), their spatial organization, and to simulate the impact of stromal deposition/degradation. B Results: b An increased deposition from natural pancreatic tissue (nPA: 2.23±0.28%) to chronic pancreatitis (iPA: 14.27±1.27%) to PC (22.30±2.031%), (p<0.0001) and a statistically significant difference in the spatial organization between nPA (1.35±0.02) vs iPA (1.70±0.01) and PC (1.73±0.01) (p<0.0001) were found. [Extracted from the article]

B Background: b Combined hepatocellular carcinoma and mass-forming cholangiocarcinoma (cHCC-MFCCC) is a rare tumor. B Conclusion: b Patients with cHCC-MFCCC showed similar rates of OS and DFS to those patients with classic HCC and MFCCC. Further evaluations of differences in tumor features and biology are necessary to better characterize the prognosis of patients with cHCC-MFCCC. [Extracted from the article]