CpG-island methylation study of liver fluke-related cholangiocarcinoma

Background: Genetic changes have been widely reported in association with cholangiocarcinoma (CCA), while epigenetic changes are poorly characterised. We aimed to further evaluate CpG-island hypermethylation in CCA at candidate loci, which may have potential as diagnostic or prognostic biomarkers. Methods: We analysed methylation of 26 CpG-islands in 102 liver fluke related-CCA and 29 adjacent normal samples using methylation-specific PCR (MSP). Methylation of interest loci was confirmed using pyrosequencing and/or combined bisulfite restriction analysis, and protein expression by immunohistochemistry. Results: A number of CpG-islands (OPCML, SFRP1, HIC1, PTEN and DcR1) showed frequency of hypermethylation in >28% of CCA, but not adjacent normal tissues. The results showed that 91% of CCA were methylated in at least one CpG-island. The OPCML was the most frequently methylated locus (72.5%) and was more frequently methylated in less differentiated CCA. Patients with methylated DcR1 had significantly longer overall survival (Median; 41.7 vs 21.7 weeks, P=0.027). Low-protein expression was found in >70% of CCA with methylation of OPCML or DcR1. Conclusion: Aberrant hypermethylation of certain loci is a common event in liver fluke-related CCA and may potentially contribute to cholangiocarcinogenesis. The OPCML and DcR1 might serve as methylation biomarkers in CCA that can be readily examined by MSP

Expert consensus document:Cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA)

Cholangiocarcinoma (CCA) is a heterogeneous group of malignancies with features of biliary tract differentiation. CCA is the second most common primary liver tumour and the incidence is increasing worldwide. CCA has high mortality owing to its aggressiveness, late diagnosis and refractory nature. In May 2015, the "European Network for the Study of Cholangiocarcinoma" (ENS-CCA: www.enscca.org or www.cholangiocarcinoma.eu) was created to promote and boost international research collaboration on the study of CCA at basic, translational and clinical level. In this Consensus Statement, we aim to provide valuable information on classifications, pathological features, risk factors, cells of origin, genetic and epigenetic modifications and current therapies available for this cancer. Moreover, future directions on basic and clinical investigations and plans for the ENS-CCA are highlighted

Influence of Operating Conditions and Physical Properties of Liquid Medium on Volumetric Oxygen Transfer Coefficient in a Dual Impeller Bioreactor

The objective of this research was to study the effects of process variables on the volumetric mass transfer coefficient of oxygen, KLa, in a stirred bioreactor using the static gassing-out method. In this study, various process conditions were chosen, including 3 parameters, namely, concentration of glucose in medium (10, 15 and 20 g/l), air flow rate (1, 1.25, 1.5 and 1.75 vvm), and agitation rate (300, 400, 500 and 600 rpm). From the results, it was found that the KLa increased with increasing air flow rate and/or speed of agitation, but decreased with increasing concentration of glucose in medium. The maximum KLa occurred when the concentration of glucose in medium was the least (10 g/l), with an air flow rate of 1.75 vvm, and an agitation rate of 600 rpm. Correlations have been developed for the estimation of volumetric mass transfer coefficients at various process conditions for medium with different glucose concentrations. The exponent values representing dependence of KLa on the process conditions were then compared with literature values

Mathematical Modeling of Drying Kinetics of Bird’s Eye Chilies in a Convective Hot-Air Dryer

The drying kinetics of red bird’s eye chilies and the color of the product were investigated in a laboratory scale hot-air dryer under 3 air temperatures of 55, 60 and 65 °C. The 6 mathematical models (Lewis model; Page model; Henderson and Pabis model; Logarithmic model; Modified Page model; and Wang and Singh model) were used to fit the experimental data obtained in order to estimate the moisture ratio as the function of drying time. The results showed that operating temperature enhanced the kinetics of the drying of chilies; the drying times of chilies at 55, 60 and 65°C were 510, 360 and 330 min, respectively. The experimental drying curves obtained at all operating conditions took place in the falling rate period. Comparing the dried products, it was observed that the red bird’s eye chilies dried at a lower temperature had higher Hunter L (lightness), a* (redness) and b* (yellowness) values. The experimental data were fitted to different drying models. The performance of these models was investigated by comparing the determination of coefficient (R2) and root mean square error (RMSE) between the observed and predicted moisture ratios. Among the 6 mathematical models, the Wang and Singh model satisfactorily described the drying kinetics of chilies. doi:10.14456/WJST.2015.1

Influence of Different Additives at Various Contents on the Properties of Pottery Clay Body

The preparation of clay body is a highly important step during the production of local decorative pottery, especially the local unglazed low-fired terracotta. In this study, sand (size < 178 mm) and Bulrush pulp were employed as an additive blended with clay (size < 125 mm) in the preparation of clay body. Various sand contents (2 - 18 wt%) and pulp contents (1 - 14 wt%) were tested. The properties of each clay body were evaluated in terms of shrinkage, water absorption and modulus of rupture. The results obtained showed that higher sand or pulp content gave higher water absorption and lower shrinkage of clay body. The dried MOR value of pulp-clay body was much higher than that of sand-clay body at the same content. The maximum fired MOR value of sand-clay body (84.51 kgf/cm2) at sand content of 2 wt% was a little higher than that of pulp-clay body (80.44 kgf/cm2) at pulp content of 4 wt%. However, at a sand content ranging between 2 and 14 % or a pulp content ranging between 1 and 5 %, the fired MOR values of clay bodies were enhanced above that of local pottery clay body (60.14 kgf/cm2). From these results the pulp could adequately be used as an additive as well. Finally, a suitable sand or pulp content for producing smooth clay bodies was between 2 and 14 wt% or between 1 and 4 wt%, respectively, due to clay bodies obtained having more smooth texture and higher strength than local pottery clay body

Serum cell-free DNA methylation of OPCML and HOXD9 as a biomarker that may aid in differential diagnosis between cholangiocarcinoma and other biliary diseases

Abstract Background Cholangiocarcinoma (CCA) is a fatal cancer of the bile duct epithelial cell lining. The misdiagnosis of CCA and other biliary diseases may occur due to the similarity of clinical manifestations and blood tests resulting in inappropriate or delayed treatment. Thus, an accurate and less-invasive method for differentiating CCA from other biliary diseases is inevitable. Methods We quantified methylation of OPCML, HOXA9, and HOXD9 in serum cell-free DNA (cfDNA) of CCA patients and other biliary diseases using methylation-sensitive high-resolution melting (MS-HRM). Their potency as differential biomarkers between CCA and other biliary diseases was also evaluated by using receiver operating characteristic (ROC) curves. Results The significant difference of methylation levels of OPCML and HOXD9 was observed in serum cfDNA of CCA compared to other biliary diseases. Assessment of serum cfDNA methylation of OPCML and HOXD9 as differential biomarkers of CCA and other biliary diseases showed the area under curve (AUC) of 0.850 (0.759–0.941) for OPCML which sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 80.00%, 90.00%, 88.88%, 81.81%, and 85.00%, respectively. The AUC of HOXD9 was 0.789 (0.686–0.892) with sensitivity, specificity, PPV, NPV, and accuracy of 67.50%, 90.00%, 87.09%, 73.46%, and 78.75%, respectively. The combined marker between OPCML and HOXD9 showed sensitivity, specificity, PPV, and NPV of 62.50%, 100%, 100%, and 72.72%, respectively, which may be helpful to prevent a misdiagnosis between CCA and other biliary diseases. Conclusions Our findings suggest the application of serum cfDNA methylation of OPCML and HOXD9 for differential diagnosis of CCA and other biliary diseases due to its less invasiveness and clinically practical method which may benefit the patients by preventing the misdiagnosis of CCA and avoiding unnecessary surgical intervention

Biomimetic scaffolds and dynamic compression enhance the properties of chondrocyte- and MSC-based tissue-engineered cartilage

Adult chondrocytes are surrounded by a protein- and glycosaminoglycan-rich extracellular matrix and are subjected to dynamic mechanical compression during daily activities. The extracellular matrix and mechanical stimuli play an important role in chondrocyte biosynthesis and homeostasis. In this study, we aimed to develop scaffold and compressive loading conditions that mimic the native cartilage micro-environment and enable enhanced chondrogenesis for tissue engineering applications. Towards this aim, we fabricated porous scaffolds based on silk fibroin (SF) and SF with gelatin/chondroitin sulfate/hyaluronate (SF-GCH), seeded the scaffolds with either human bone marrow mesenchymal stromal cells (BM-MSCs) or chondrocytes, and evaluated their performance with and without dynamic compression. Human chondrocytes derived from osteoarthritic joints and BM-MSCs were seeded in scaffolds, precultured for 1 week, and subjected to compression with 10% dynamic strain at 1 Hz, 1 hr/day for 2 weeks. When dynamic compression was applied, chondrocytes significantly increased expression of aggrecan (ACAN) and collagen X (COL10A1) up to fivefold higher than free-swelling controls. In addition, dynamic compression dramatically improved the chondrogenesis and chondrocyte biosynthesis cultured in both SF and SF-GCH scaffolds evidenced by glycosaminoglycan (GAG) content, GAG/DNA ratio, and immunostaining of collagen type II and aggrecan. However, both chondrocytes and BM-MSCs cultured in SF-GCH scaffolds under dynamic compression showed higher GAG content and compressive modulus than those in SF scaffolds. In conclusion, the micro-environment provided by SF-GCH scaffolds and dynamic compression enhances chondrocyte biosynthesis and matrix accumulation, indicating their potential for cartilage tissue engineering applications