Mcmanuslamont5472

© 2020 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc.Cholangiocytes secrete bicarbonate and absorb glucose, producing bile with alkaline pH and low glucose content. These functions of cholangiocytes have been suggested as a marker of bile duct viability during normothermic ex situ liver perfusion (NESLiP), and are now monitored routinely post reperfusion in our centre. In this study, we reviewed the composition of bile immediately after reperfusion in liver transplant recipients to determine normal post-transplant parameters and the predictive value of bile biochemistry for the later development of cholangiopathy. After reperfusion of the liver graft, a cannula was placed in the bile duct to collect bile over a median 44 minute time period. The bile produced was analysed using a point of care blood gas analyser (Roche Cobas b221). 100 liver transplants (35 donation after circulatory death [DCD] and 65 donation after brain death [DBD]) were studied. Median bile pH was 7.82 (Interquartile range [IQR] 7.67 -7.98), median bile glucose 2.1 (1.4 - 3.7) mmol/L, median blood and bile pH difference 0.5 (0.37 - 0.62) and the median blood and bile glucose difference 7.1 (5.6 - 9.1) mmol/L. 12 recipients developed cholangiopathy over a median follow up of 15 months (IQR 11 - 20). Bile sodium (142 vs 147 mmol/L; p=0.02) and blood-bile glucose concentration differences (5.2 vs 7.6; p=0.001) were significantly lower and associated with ischaemic cholangiopathy. In conclusion, bile biochemistry may provide useful insights into cholangiocyte function, and hence bile duct viability. Our results suggest bile glucose is the most sensitive predictor of cholangiopathy. This article is protected by copyright. All rights reserved.Platelets are most important to provide cellular hemostasis but also take part in inflammation and immune processes. Increased platelet size has been regarded as a feature describing a young and more reactive subpopulation until studies were published which questioned this concept. Changes of platelet size given by the mean platelet volume (MPV) were described for immune thrombocytopenia, cardiovascular disease, atherosclerosis, venous thromboembolism, chronic lung disease, sepsis, cancer associated thrombosis, autoimmune disorders and others. This review summarizes the literature on what is known about platelets with different size and describes controversies of studies with large and small platelets putting a focus on their thrombogenicity and age in functional studies and on the association of MPV with the mentioned diseases. This article is protected by copyright. All rights reserved.Collagen biomaterials are widely used for soft tissue augmentation. Cross-linking techniques for collagen matrix (CM) achieve mechanical and volumetric stability; nevertheless, cross-linking may compromise biocompatibility. The aim of the present study was to investigate two different three-dimensional (3D) porcine-derived CMs, noncross-linked (ncl)_CM and cross-linked (cl)_CM, for their effects on macrophages (Mφ) and gingival fibroblasts. The effects of the CMs on the cell viability, proliferation, and polarization of Mφ derived from human monocyte THP-1 cells were assessed. The effects of paracrine factors from Mφ cultured on the CMs were further studied in human gingival fibroblasts (HGF-1 cells). The spongy layer of ncl_CM was partially resorbed after 1 day of culture. cl_CM maintained increased numbers of viable cells when compared with ncl_CM on day 3 for both THP-1 and HGF-1 cells. Higher mRNA levels of M1 markers, including IL-1 and IL-6, were found in Mφ cultured on cl_CM, while no significant differences were observed in M2 marker expression levels, including Arg1 and CD206, for cells cultured on both CMs when compared with those of the control. Furthermore, the conditioned medium collected from Mφ cultured on both CMs decreased cell viability. Nevertheless, neither of the CM-conditioned media influenced the mRNA levels of TGF-β, COL1a2, and PDGF-A in HGF-1 cells when compared with the control media. A comparison showed that cl_CM tended to result in more viable cells than ncl_CM, while cl_CM polarized Mφ toward an M1 phenotype, which was confirmed by the observation of increased mRNA levels of pro-inflammatory cytokines. © 2020 Wiley Periodicals, Inc.Multidrug resistance is one of the reasons for low survival of advanced hepatocellular carcinoma (HCC). Our previous studies indicate that the hedgehog signalling is involved in hepatic carcinogenesis, metastasis and chemo-resistance. The present study aims to uncover molecular mechanisms underlying hepatoma chemo-resistance. TAP1 and GLI1/2 gene expression was assessed in both poorly differentiated hepatoma cells and HCC specimens. Potential GLI-binding site in the TAP1 promoter sequence was validated by molecular assays. Approximately 75% HCC specimens exhibited an elevated expression of hedgehog GLI1 transcription factor compared with adjacent liver tissue. Both GLI1/2 and TAP1 protein levels were significantly elevated in poorly differentiated hepatoma cells. Both Huh-7-trans and Huh-7-DN displayed more karyotypic abnormalities and differential gene expression profiles than their native Huh-7 cells. Sensitivity to Sorafenib, doxorubicin and cisplatin was remarkably improved after either GLI1 or TAP1 gene was inhibited by an RNAi approach or by a specific GLI1/2 inhibitor, GANT61. Further experiments confirmed that hedgehog transcription factor GLI1/2 binds to the TAP1 promoter, indicating that TAP1 is one of GLI1/2 target genes. In conclusion, TAP1 is under direct transcriptional control of the hedgehog signalling. Targeting hedgehog signalling confers a novel insight into alleviating drug resistance in the treatment of refractory HCC. © 2020 The Authors. Journal of Cellular and Molecular Medicine published by by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.BACKGROUND Previous studies have demonstrated that the A1A2A3 domains of VWF play a key role in regulating macrophage-mediated clearance in-vivo. In particular, the A1-domain has been shown to modulate interaction with macrophage LRP1 clearance receptor. selleckchem Furthermore, N-linked glycans within the A2-domain have been shown to protect VWF against premature LRP1-mediated clearance. Importantly however, the specific regions within A1A2A3 that enable macrophage binding have not been defined. OBJECTIVE AND METHODS To address this, we utilised site-directed PEGylation and introduced novel targeted N-linked glycosylation within A1A2A3-VWF and subsequently examined VWF clearance. RESULTS Conjugation with a 40-kDa PEG moiety significantly extended the half-life of A1A2A3-VWF in VWF-/- mice in a site-specific manner. For example, PEGylation at specific sites within the A1-domain (S1286) and A3-domain (V1803, S1807) attenuated VWF clearance in-vivo, compared to wild-type A1A2A3-VWF. Furthermore, PEGylation at these specific sites ablated binding to differentiated THP-1 macrophages and LRP1 cluster II and cluster IV in-vitro.