Distinct miRNA signatures associate with subtypes of cholangiocarcinoma from infection with the tumourigenic liver fluke Opisthorchis viverrini

Background & Aims: Intrahepatic cholangiocarcinoma (ICC) is a significant public health problem in East Asia, where it is strongly associated with chronic infection by the food-borne parasite Opisthorchis viverrini (OV). We report the first comprehensive miRNA expression profiling by microarray of the most common histologic grades and subtypes of ICC: well differentiated, moderately differentiated, and papillary ICC. Methods: MicroRNA expression profiles from FFPE were compared among the following: ICC tumour tissue (n = 16), nontumour tissue distally macrodissected from the same ICC tumour block (n = 15), and normal tissue (n = 13) from individuals undergoing gastric bypass surgery. A panel of deregulated miRNAs was validated by qPCR. Results: Each histologic grade and subtype of ICC displayed a distinct miRNA profile, with no cohort of miRNAs emerging as commonly deregulated. Moderately differentiated ICC showed the greatest miRNA deregulation in quantity and magnitude, followed by the papillary subtype, and then well differentiated ICC. Moreover, when ICC tumour tissues were compared to adjacent non-tumour tissue, similar miRNA dysregulation profiles were observed. Conclusions: We show that common histologic grades and subtypes of ICC have distinct miRNA profiles. As histological grade and subtypes are associated with ICC aggressiveness, these profiles could be used to enhance the early detection and improve the personalised treatment for ICC. These findings also suggest the involvement of specific miRNAs during ICC tumour progression and differentiation. We plan to use these insights to (a) detect these profiles in circulation and (b) conduct functional analyses to decipher the roles of miRNAs in ICC tumour differentiation

The miRNAome of Opisthorchis viverrini induced intrahepatic cholangiocarcinoma

Intrahepatic cholangiocarcinoma (ICC) is an aggressive cancer, arising in the biliary ducts that extend into the liver. The highest incidence of ICC occurs in Southeast Asia, particularly in the Mekong River Basin countries of Thailand, Laos, Cambodia, and Vietnam, where it is strongly associated with chronic infection by the food-borne liver fluke Opisthorchis viverrini (OV), one of only three eukaryote pathogens considered Group one carcinogens. Intrahepatic cholangiocarcinoma is usually diagnosed at an advanced stage, with a poor prognosis and survival often less than 24 months. Hence, biomarkers that enable the early detection of ICC would be desirable and have a potentially important impact on the public health in the resource-poor regions where this cancer is most prevalent. As microRNAs (miRNAs) remain well preserved after formalin fixation, there is much interest in developing them as biomarkers that can be investigated using tumor biopsy samples preserved in formalin fixed paraffin embedded (FFPE) tumor blocks. Recently, we reported the first comprehensive profiling of tissue-based miRNA expression using FFPE from the three most common subtypes of OV-induced ICC tumors: moderately differentiated ICC, papillary ICC, and well-differentiated ICC. We observed that each subtype of OV-induced ICC exhibited a distinct miRNA profile, which suggested the involvement of specific sets of miRNAs in the progression of this cancer. In addition, non-tumor tissue adjacent to ICC tumor tissue on the same FFPE block shared a similar miRNA dysregulation profile with the tumor tissue than with normal (non-tumor) liver tissue (individuals without ICC or OV infection). Herein, we provide a detailed description of the microarray analysis procedures used to derive these findings

Elevated Plasma IL-6 Associates with Increased Risk of Advanced Fibrosis and Cholangiocarcinoma in Individuals Infected by Opisthorchis viverrini

Opisthorchis viverrini is considered among the most important of the food-borne trematodes due to its strong association with advanced periductal fibrosis and bile duct cancer (cholangiocarcinoma). We investigated the relationship between plasma levels of Interleukin (IL)-6 and the risk of developing advanced fibrosis and bile duct cancer from chronic Opisthorchis infection. We show that IL-6 circulates in plasma at concentrations 58 times higher in individuals with advanced fibrosis than age, sex, and nearest-neighbor matched controls and 221 times higher in individuals with bile duct cancer than controls. We also observed a dose-response relationship between increasing levels of plasma IL-6 and increasing risk of advanced fibrosis and bile duct cancer; for example, in age and sex adjusted analyses, individuals with the highest quartiles of plasma IL-6 had a 19 times greater risk of developing advanced periductal fibrosis and a 150 times greater risk of developing of bile duct cancer than individuals with no detectable level of plasma IL-6. Finally, we show that a single plasma IL-6 measurement has excellent positive predictive value for the detection of both advanced bile duct fibrosis and bile duct cancer in regions with high O. viverrini transmission. These data support our hypothesis that common mechanisms drive bile duct fibrosis and bile duct tumorogenesis from chronic O. viverrini infection. Our study also adds a unique aspect to the literature on circulating levels of IL-6 as an immune marker of hepatobiliary pathology by showing that high levels of circulating IL-6 in plasma are not related to infection with O. viverrini, but to the development of the advanced and often lethal pathologies resulting from chronic O. viverrini infection

Up-regulation of annexin A2 in cholangiocarcinoma caused by Opisthorchis viverrini and its implication as a prognostic marker

Cholangiocarcinoma (CCA), or cancer of the bile ducts, is primarily associated with infection with the liver fluke Opisthorchis viverrini in northeast Thailand. The disease is associated with late presentation, poses challenges for diagnosis and has a high mortality rate--features that highlight the need for tumor markers. At present, there are no specific tumor markers that can indicate the early stages and status of CCA. Proteomic analysis of the proteins expressed on the surface of tumor cells is particularly difficult since proteome-wide analysis of surface membrane proteins has thus far been hampered by the lack of effective strategies to profile hydrophobic membrane proteins. In this study, a sequential protein extraction was utilized to overcome this problem. Membrane protein was extracted from four CCA cell lines with different tumor forming capabilities. The non-tumor H69 biliary cell line was used as a control. Two-dimensional-PAGE followed by MALDI-TOF-MS was used to identify differentially expressed proteins. Among 20 up-regulated membrane proteins identified in the CCA cell lines was ANXA2, a participant in tumor invasion and metastasis in other cancers. Accordingly, ANXA2 was verified in human subjects by probing, using a commercial anti-mouse monoclonal antibody and a tissue microarray of CCA (301 diagnosed cases), where it was found to associate with one of several tumor progression stages as reflected by lymphatic invasion (P=0.014) and metastasis (P=0.026). Patients with high expression of ANXA2 had a significantly shorter survival time (P=0.011). ANXA2 expression in tumors may be useful for predicting the poor outcome of CCA patients

Cost Analysis of Physical Therapy Clinic In Thailand: The impact of the COVID-19 outbreak

Objective: The aim of the study was to identify the financial burden and compare the unit costs that Coronavirus disease 2019 (COVID-19) imposed on a physical therapy clinic that are one of the primary forms of healthcare facilities in Thailand.Design and Setting: The study was a retrospective study conducted at the physical therapy clinic, Khon Kaen University Community Outreach Center (KKUCOMOC) in Thailand. To assess the impact of the outbreak, the service unit was divided into two units: physical therapy (PT) and Thai massage (TM), and the annual report for the 2019-2021 fiscal year was analyzed as representative of before and during an outbreak. The study tool was the Handbook of Unit Cost Analysis from the Office of the Permanent Secretary. Revenue, expenses, and the number of patient visits were all gathered. The data was then analyzed and summarized using Microsoft Excel programs, and descriptive statistics were presented. Results: The COVID-19 pandemic not only reduced the number of patient visits but also the revenue. During the pandemic, the percentage of expense to the facility’s revenue increased in both units. Labor, material, and capital costs were the major cost components, with labor costs accounting for the majority of direct costs. The first emerged in the 2020 fiscal year, resulting in an increase in PT's and TM's unit costs of 26.66% ($US10.24 to$US12.97) and 22.69% ($US6.92 to$US8.49), respectively. Following that, it continued to rise in both units throughout the subsequent fiscal year. As a result, at the end of the study, the unit costs of PT and TM were $US13.95 and$US9.22 respectively. Conclusions: The COVID-19 pandemic reduced the number of patient visits and revenue. Furthermore, it raised the unit cost of PT and TM to $US13.95 and$US9.22 respectively

Segmental Assessment of Trunk Control in Moderate-to-Late Preterm Infants Related to Sitting Development

(1) Background: The assessment of postural segment control in premature infants seems to be critical during the onset of upright gross motor development, especially sitting. Identifying correlations between postural segment control and the development of sitting milestones could help with promoting optimal gross motor movement. However, data on this topic in home-raised premature infants via longitudinal design are still limited. The purpose of this study was to examine relationships between postural segment control and sitting development through series assessments from the corrected age of 4 months until the early onset of independent sitting attainment. (2) Methods: 33 moderate-to-late premature infants were recruited. Their trunk segment control was assessed using the Segmental Assessment of Trunk Control (SATCo), and sitting development was examined by the Alberta Infant Motor Scale (AIMS). Relationships between SATCo and sitting scores were analysed using Spearman’s rank correlation (rs). (3) Results: significant fair-to-good correlations between segmental trunk control and sitting scales were found from 4 months (rs = 0.370–0.420, p < 0.05) to the age of independent sitting attainment (rs = 0.561–0.602, p < 0.01). (4) Conclusion: relationships between the trunk segment control and sitting ability of moderate-to-late preterm infants were increased in accordance with age

Banking on the future: Biobanking for omics approaches to biomarker discovery for Opisthorchis-induced cholangiocarcinoma in Thailand

Cholangiocarcinoma (CCA) – bile duct cancer - is associated with late presentation, poses challenges for diagnosis, and has high mortality. These features t highlight the desperate need for biomarkers than can be measured early and in accessible body fluids such as plasma of people at risk for developing this lethal cancer. In this manuscript, we address previous limitations in the discovery stage of biomarker(s) for CCA and indicate how new generation of “omics’ technologies could be used for biomarker discovery in Thailand. A key factor in the success of this biomarker program for CCA is the combination of cutting edge technology with strategic sample acquisition by a biorepositories