FULL-TEXT ARTICLES FROM ORAL PRESENTATIONS Transcriptomic Meta-Analysis in Pancreatic Ductal Reveals Therapeutic Targets and Diagnostic Biomarkers

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is the most common form of pancreatic cancer, which has the highest mortality rate of all solid tumors. The absence of an effective screening process and distinctive symptoms causes a delay in diagnosis. Traditional chemotherapy and curative surgery have limited benefits on patient survival. Enzymes are one of the most important groups of drug targets and are preferred markers for the detection of various diseases. OBJECTIVES: This study aims to identify up-regulated genes encoding enzymes in PDAC to suggest novel therapeutic targets for more effective treatments to be developed and diagnostic biomarkers for PDAC. MATERIALS and METHODS: NCBI Gene Expression Omnibus (GEO) was searched for datasets using keywords ‘pancreatic ductal adenocarcinoma’. The inclusion criteria were i) Gene expression microarray data, ii) human-derived pancreatic ductal adenocarcinoma tissues and normal pancreatic tissue samples. All data processing and integration procedures were performed using ExAtlas. The false discovery rate is less than 0.05, and the change of gene expression is ≥ 10-fold were considered significant. The up-regulated enzyme-coding genes were detected in the differentially expressed gene list. The identified up-regulation of enzyme-coding genes in PDAC was verified using datasets from TCGA and GTEx projects. RESULTS: The random effect integrative meta-analysis of five submissions (GSE46234, GSE19280, GSE43795, GSE41368, and GSE71989) containing 24 tumor-normal tissue pairs revealed 22 up-regulated genes, two of which encoding enzymes. The enzyme-coding genes with at least 10-fold differential expression compared to the controls were SULF1 (sulfatase, fold change=22.135) and KYNU (kynureninase, fold change=10.716), in consistence with the results from TCGA and GTEx data. CONCLUSIONS: The results of this study suggest that sulfatase and kynureninase may have the potential to become diagnostic biomarkers and therapeutic targets for PDAC, which merits further investigation. Abstract: In the Republic of North Macedonia the work of the diagnostic medical laboratories is regulated by the Law of Health Care. There is an urgent need for better development of an evidence-based, scientific, and sustainable national strategy for the improvement of health laboratory service. Clear indicators of improvement have to be established. A key indicator should be the number of laboratories that have achieved and can maintain accreditation. The Macedonian Society of Medical Biochemistry and Laboratory Medicine (MSMBLM) recommends that the quality system established meets the requirements of the International Standard for medical laboratories (‘Medical laboratories: Requirements for quality and competence [EN ISO 15189:2012]. The accreditation of Macedonian medical laboratories is not mandatory; the decision for accreditation is voluntary. So far, nine medical laboratories have been accredited according the MKS EN ISO 15189:2013. Four of them are public sector laboratories. The small number of accredited laboratories could be the result of the shortage of financial resources, poor government attention to laboratory service, the shortage of qualified personnel and/or the lack of a national laboratory policy. The experiences of laboratory professionals from accredited laboratories, who have a high level of knowledge, skills, and competence, are crucially important to the process of developing a competent laboratory service within the national health system. ABSTRACT Objectives: In this study, we aimed to determine the effect of bilberry tea samples on the markers of the intrinsic and extrinsic pathways of apoptosis in the HCT116 colon cancer cell line. Materials and Methods: Bilberry tea in different infusions and boiling periods (1 min, 3 min, 5 min, 10 min) were prepared and phenolic levels were determined by LC MS / MS technique. The highest phenolic content was determined in tea samples of dried shredded fruit for 5 min boiling, so this product was chosen for in vitro study. Cytotoxicity and viability tests were performed by adding WST-8 solution. Intrinsic and extrinsic pathways of apoptosis were assessed by determining the TNF-related apoptosis-inducing ligand (TRAIL), Apoptotic Protease Activating Factor-1 (APAF-1), Cytochrome-c, Caspase -3,-8,-9 levels in HCT-116 colon cancer cell line. Results: Cytotoxicity studies in cell culture were conducted using 50-10 µg/ml of bilberry tea samples which was prepared at a concentration of 5 g/10 ml. The levels of Caspase 3, APAF-1, TRAIL and Cytochrome-c were significantly higher in bilberry added cell culture than the control cells. Other markers (caspase -8, -9 levels) did not show any significant change compared to control cells. Conclusions: It is concluded that bilberries induced TRAIL, APAF-1, Caspase-3, Cytochrome-c and consequently induced both intrinsic and extrinsic pathways of apoptosis. on HCT-116 colon cancer cell line was assessed. The studies with natural compunds have shown that the different solvents, and the methods used to dissolve the plant or the fruit affects the study results related to active compounds in the plant or fruit studied. In the studies of Yildiz et al. (2011) related to Vaccinium myrtillus L., the structure of the biological active components of Vaccinium myrtillus L. were clarified, and, the collected fruits were stored at -18˚C for 1 week, and then dried at 40˚C and pulverized. Dried fruits were extracted with methanol, ethanol, water and ethyl acetate, and left to stand in the stirrers with heater for 24 hours. The analyzes were performed by RC-HPLC. Researchers investigating the amount of phenolic compounds have found that the pehnolic content of plants show high variability due to the factors such as location, growing conditions, soil properties, irrigation, temperature and sunbathing. Previously, caffeic acid (6.29 ± 3.55 µg/g), epicatechin (8.35 ± 5.04 µg/g), and myricetin (4.16 ± 1.81 µg/g), were found as the highest phenolic molecules in bilberry extracts (2.10 ± 0.64 µg/g) (19). The most promising anticarcinogenic agents in plants are these phenolic compounds, which are abundantly present in Bilberries (Vaccinium myrtillus) (20). In these studies, the bilberry fruits were extracted in the different types of solvents (water, ethanol and ethyl acetate), and their chemical compositions were identified using HPLC - DAD and HPLC-MS/MS method and the effects on antioxidant system were investigated using in-vitro analysis. In the result of these experimental studies, antiradical and antioxidant effects were found to be higher in ethanol, and ethyl acetate extracts, as Abstract Objectives: Normal oxygen delivery is essential for survival. Hypoxia, which is a common feature of various pathological conditions, ranging from cancer to inflammatory diseases, occurs when normal oxygen delivery is altered by an im- balance between cellular oxygen demand and tissue oxygen supply. Among the intricate mechanisms organisms have developed to maintain oxygen homeosta- sis, a family of hypoxia-inducible transcription factors (HIFs), are found to be the main regulator adaptive cellular response to hypoxia. Although ELISA can be used for its measurement, the lability of the protein and length of the analysis (>5 hours) pose limitations. Thus, our aim is to develop an electrochemical impe- dance spectroscopy (EIS) based biosensor system for quick and reliable measurement of HIF-1α in serum. Materials and Methods : HIF-1alpha antibodies have been used as a biorecognition receptor. For immobilization, the electrode was first modified with albumin, followed by PAMAM. The new biosensor was compared with the conventional ELISA method. Results : Based on the chronoimpedance data, total analysis time for EIS was chosen as 15 minutes. Calibration curve was constructed by locating electron transfer resistance on y-axis and HIF1 concentration on x-axis, between 50-1000 pg/mL. LOD and LOQ of the biosensor were calculated as 14.45 pg/mL and 43.8 pg/mL, respectively. The new biosensor showed very good correlation when compared with the conventional ELISA method (R2= 0.99649). Conclusion : We developed and analytically validated a biosensor system to measure HIF-1α in serum. This new biosensor promises more timely and accurate measurements in determining the tissue oxygenation in patients who have hypoxia related conditions such as diabetic foot. target molecule HIF-1. The measurement of the binding of HIF-1 to the anti- HIF-1 on the electrode was performed by using surface-characterization EIS and the results were obtained with concentration-time-impedance function. In order to evaluate the validity of our measurement, results, procedure and performance characteristics were compared with the conventional ELISA technique. bonding properties of surface after Abstract Background: Every year more than one million people commit suicide worl- dwide. Suicide cases constitute 1-2% of the total global mortality. Reelin is an extracellular matrix glycoprotein and involved in the development of brain layers during embryogenesis. Reelin is linked with several psychiatric diseases but not investigated in the suicide cases. Objectives: We aimed to investigate reelin enzyme levels among suicide patients in comparison to the heathy controls. Materials and Methods: A total 86 suicide cases and 100 healthy controls were included in the study. Serum reelin levels were determined by using commercial Human ELISA kit. Demographic variables and clinic data were collected and analyzed. Results: Body mass index and mean age among groups were significantly dif- ferent. While the median reelin enzyme level of Suicide Group was 3038,31 (IQR:212,46-8044,21) ng/L and that of Control Group was 2271,20 (IQR:77,67- 7647,83) ng/L. The difference was statistically significant (p <0.01). Conclusions: According to the results of this study, reelin enzyme level of suici-