Demographic Change Across the Lifespan of Pet Dogs and Their Impact on Health Status

Although dogs' life expectancies are six to twelve times shorter than that of humans, the demographics (e. g., living conditions) of dogs can still change considerably with aging, similarly to humans. Despite the fact that the dog is a particularly good model for human healthspan, and the number of aged dogs in the population is growing in parallel with aged humans, there has been few previous attempts to describe demographic changes statistically. We utilized an on-line questionnaire to examine the link between the age and health of the dog, and owner and dog demographics in a cross-sectional Hungarian sample. Results from univariate analyses revealed that 20 of the 27 demographic variables measured differed significantly between six dog age groups. Our results revealed that pure breed dogs suffered from health problems at a younger age, and may die at an earlier age than mixed breeds. The oldest dog group (>12 years) consisted of fewer pure breeds than mixed breeds and the mixed breeds sample was on average older than the pure breed sample. Old dogs were classified more frequently as unhealthy, less often had a “normal” body condition score, and more often received medication and supplements. They were also more often male, neutered, suffered health problems (such as sensory, joint, and/or tooth problems), received less activity/interaction/training with the owner, and were more likely to have experienced one or more traumatic events. Surprisingly, the youngest age group contained more pure breeds, were more often fed raw meat, and had owners aged under 29 years, reflecting new trends among younger owners. The high prevalence of dogs that had experienced one or more traumatic events in their lifetime (over 40% of the sample), indicates that welfare and health could be improved by informing owners of the greatest risk factors of trauma, and providing interventions to reduce their impact. Experiencing multiple life events such as spending time in a shelter, changing owners, traumatic injury/prolonged disease/surgery, getting lost, and changes in family structure increased the likelihood that owners reported that their dogs currently show behavioral signs that they attribute to the previous trauma

Expression of Tight Junction Components in Hepatocyte-Like Cells Differentiated from Human Embryonic Stem Cells

Human embryonic stem cells can be differentiated in vitro into a wide variety of progeny cells by addition of different morphogens and growth factors. Our aim was to monitor the expression pattern of tight junction (TJ) components and various cellular markers during differentiation of stem cell lines toward the hepatic lineage. Human embryonic stem cell lines (HUES1, HUES9) were differentiated into endoderm-like cells, and further differentiated to hepatocyte-like cells. Gene expressions of Oct3/4, Nanog, alpha-fetoprotein, albumin, cytokeratins (CK-7, CK-8, CK-18, CK-19), ATP-binding cassette (ABC) transporters (ABCC2, ABCC7, ABCG2), and various TJ components, including claudin-1, claudin-4, claudin-5, claudin-7, and tricellulin, as well as an extracellular matrix component, agrin were monitored during hepatic differentiation by real-time quantitative PCR. The differentiated cells exhibit epithelial morphology and functional assessments similar to that of hepatocytes. The expression level of stem cell marker genes (Oct3/4 and Nanog) significantly and gradually decreased, while liver-associated genes (alpha-fetoprotein, albumin) reached their highest expression at the end of the differentiation. The endoderm-like cells expressed claudin-1, which declined eventually. The expression levels of cholangiocyte markers including claudin-4, CK-7, CK-19, and agrin gradually increased and reached their highest level at the final stage of differentiation. In contrast, these cells did not express notable level of claudin-7, CK-8 and tricellulin. The marker set used for monitoring differentiation revealed both hepatocyte and cholangiocyte characteristics of the differentiated cells at the final stage. This is the first report describing the expression level changes of various TJ components, and underlining their importance in hepatic differentiation. © 2015 Arányi Lajos Foundatio

Molecular Characteristics of Fibrolamellar Hepatocellular Carcinoma

Fibrolamellar hepatocellular carcinoma (FLC) occurs in non-cirrhotic liver and the etiopathogenesis is still obscure. Both hepatocellular and cholangiocellular markers are expressed in the tumor, however, molecular alterations and altered pathways playing role in the tumor pathogenesis are not clearly identified. The purpose of the present study was to compare the expression level of EGFR, syndecan-1 and ss-catenin in FLC, conventional hepatocellular carcinoma (cHCC) and cholangiocellular carcinoma (CCC) and to investigate the possibility of mutation both in EGFR and K-RAS. Eight FLCs were compared with 7 cHCCs, 7 CCCs and 5 normal liver samples. Cytokeratins 7, 8, 18, 19, HepPar1 (HSA), EGFR, syndecan-1 (CD138) and ss-catenin were detected by immunohistochemistry. In addition EGFR, ss-catenin and syndecan-1 were evaluated by digital morphometry and K-RAS, EGFR mutations in FLC cases using paraffin-embedded samples. All FLCs were positive for HepPar1 (HSA) and cytokeratins 7, 8, 18, but negative for cytokeratin 19 by immunohistochemistry. EGFR was significantly overexpressed in all three tumor types, being highest in FLCs (p = 0,0001). EGFR, K-RAS mutation analyses revealed no mutations in exons studied in FLCs. Our findings proved that expression of EGFR is higher in FLC than in other types of primary malignant hepatic tumors and no K-RAS mutation can be detected, so FLC is a good candidate for anti-EGFR treatment

Tricellulin Expression and its Prognostic Significance in Primary Liver Carcinomas.

Numerous data suggest that altered expression of tight junction proteins such as occludin and claudins plays important role in carcinogenesis. However, little is known about tricellulin, a transmembrane tight junction protein concentrated where three epithelial cells meet. We aimed to characterize tricellulin expression in normal and cirrhotic liver in comparison to primary hepatic neoplasms. Tricellulin expression of 20 control livers, 12 cirrhotic livers, 32 hepatocellular carcinomas (HCC), and 20 intrahepatic cholangiocarcinomas (iCCC) was investigated by immunohistochemistry and Western blotting. Co-localization of tricellulin with claudin-1, -4, and MRP2 was studied using double immunofluorescence. Scattered tricellulin immunopositivity was restricted to biliary pole of hepatocytes confirmed by co-localization with MRP2. Moreover, spotted-like reaction was observed between bile duct epithelial cells. In 40 % of HCCs marked tricellulin overexpression was measured regardless of tumor grades. In iCCCs, however, tricellulin expression decreased parallel with dedifferentiation. In HCCs high tricellulin expression, in iCCCs low tricellulin expression correlated with poor prognosis. Co-localization with MRP2 might substantiate that tricellulin plays role in blood-biliary barrier. Overexpressed tricellulin in a subset of HCCs correlated with unfavorable prognosis. Similar to ductal pancreatic adenocarcinoma, higher grades of iCCCs were associated with decreased tricellulin expression correlating with poor prognosis