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International Conference on Antioxidants and Diseases ICADD 2018 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 International Conference on Antioxidants and Diseases ICADD 2018 "New Frontiers in Omics and Redox Medicine" 18 - 19th July 2018 Pullman Kuala Lumpur City Centre 1 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 PLENARY 1 Omics, Inflammation and Gut RAJA AFFENDI RAJA ALI Gastroenterology Unit, Department of Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia (Email: [email protected]) Gut health influences the general well-being of human as it provides the core body’s immune system. Environmental factors (exposome) such as diet, nutrients, smoking, obesity may interact with the intestinal microenvironment leading to multiple pathologies inside the gut. An inappropriate immune response towards environmental changes directing to oxidative stress in the mucosal layer of the GI tract. This will subsequently lead to inflammation and destruction in the intestinal mucosal. Inflammatory bowel disease (IBD), which is increasing steadily in the Asia pacific region, is a good example to explain these processes. In recent years, with the help of new high throughput technologies have enabled a better understanding on the genetic susceptibility of patients with IBD that interacts with the gut microbiota. The host-microbiota response will trigger the cell mediated inflammation causing the released of multiple cytokines. We have conducted transcriptomic profiling to understand the changes in the transcriptomes of long duration as compared to the short duration IBD patients. Differentially expressed genes include genes that are related with production of reactive oxygen species. Among the signaling pathway identified in this study was fatty acid metabolism, which is closely related to inflammation and IBD-related cancer (up to 20% of cancers arise from chronic inflammation). In daily routine, short chain fatty acids are produced by gut microbiota through fermentation of dietary fibers and resistant starch. Short chain fatty acids produced in colon may affect pro-inflammatory cytokines and enhance the migration of neutrophils to the inflammation site and helps in the process of phagocytosis. Our team has also shown the beneficial effects of modulating the gut immunity in post-operative patients with colorectal cancer as well as in patients with irritable bowel syndrome. With remarkable advanced in omics technology has reshaped the way we tackle gastrointestinal diseases. 2 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 PLENARY 2 Neuroinflammatory Mechanisms Involved in Alzheimer’s Disease DOUGLAS GORDON WALKER Molecular Neuroscience Research Center, Shiga University of Medical Science, Seta, Otsu, Japan (Email: [email protected]) Inflammation has been considered as a pathological mechanism for Alzheimer’s disease (AD) for more than 30 years, initially based on the pathological observations of activated microglia identified in postmortem human brains. In recent years, modern gene expression profiling techniques and single nucleotide polymorphism screening have been applied to identify different inflammation associated changes and new proteins involved. There have been discrepancies in the two approaches that need to be considered. Ultimately, the goals of all of these studies are to identify inflammation associated therapeutic targets for AD, but there is still incomplete understanding of how inflammation affects AD and the best way to modulate it. One approach is address the question ‘what are microglia in AD brains actually doing”. Comparison will be made between results from publicly available gene databases of AD and control human brain tissue, and immunohistochemical approaches using human brain tissues to phenotype microglial expression of key target proteins. Gene expression databases from human materials provide a wealth of information about inflammatory changes in AD brains with results conflicting with established concepts. To address the fundamental question about the extent and localization of activated microglia, we will consider the distribution of microglia expressing the purinergic receptor P2RY12 in relation to AD pathological structures. P2RY12 is considered a marker for non-activated microglia. In addition, genetic studies have linked triggering receptor expressed by myeloid cells-2 (TREM-2), CD33 and progranulin with inflammatory mechanisms of neurodegeneration. The expression of these proteins by microglia and their known biological functions will be considered in the context of AD inflammatory mechanisms. To understand and develop treatments for AD, an accurate picture of what is happening in human disease affected tissues is needed. This is needed to ensure that animal models used for testing therapies accurately reflect the human disease. Keywords: Microglia, immunohistochemistry, gene expression, phenotypes, neurodegeneration 3 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 PLENARY 3 Is H2O2 Production Increased at High Mitochondrial Membrane Potential? Facts and Artefacts under Normoxia, Hyperoxia, and Hypoxia. KOMLÒDI T1, SOBOTKA O2, DOERRIER C1, GNAIGER E1,3 1Oroboros Instruments, Innsbruck, Austria, 2Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Czech Republic, 3D. Swarovski Research Laboratory, Department of Visceral, Transplant and Thoracic Surgery, Medical University of Innsbruck, Austria (Email: [email protected]) Hydrogen peroxide (H2O2) production increases at high mitochondrial membrane potential in the resting state of LEAK-respiration relative to the ADP-activated OXPHOS state. This OXPHOS-LEAK paradigm of mitochondrial ROS production, however, is based on studies which typically ignore the fact that intracellular oxygen levels in the microenvironment of mitochondria are ‘hypoxic’ relative to environmental normoxia. Can the OXPHOS-LEAK-ROS paradigm be extended to physiological intracellular oxygen conditions? The Amplex UltraRed (AmR) assay is one of the most popular fluorescence methods for determination of mitochondrial and cellular net H2O2 flux. Quantitative and accurate measurement of ROS production, however, remains a critical issue. Quality control is required to avoid numerous artefacts. Independent of H2O2 flux, increasing fluorescence signals may result from photosensitive auto-oxidation of AmR, which represents a confounding chemical background process. What is fact and artefact in the fluorometric AmR assay under normoxia and hypoxia? H2O2 production and oxygen consumption were measured simultaneously in mitochondria isolated from mouse brain and heart, and in intact cells of Saccharomyces cerevisiae, using the O2k-FluoRespirometer (Oroboros Instruments, Austria) and four respiration media: Dulbecco‘s phosphate buffer saline (DBPS); KCl-based medium; respiratory media MiR05-Kit and Buffer z. Oxygen flux remained independent of oxygen concentration under normoxia and ‘hypoxia’. In contrast, apparent H2O2 production increased significantly during repeated hypoxia/anoxia transitions of yeast suspended in DPBS. Such hypoxic peaks of H2O2 production, however, were revealed as an artefact of auto-oxidation in DBPS and KCl-based media, not observed in MiR05 and Buffer z. The actual H2O2 flux declined from normoxia to hypoxia and anoxia in intact yeast and isolated mitochondria in the active OXPHOS state stimulated by ADP, whereas H2O2 flux was constant from normoxia to hypoxia in the LEAK state in the absence 4 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 of stimulation by ADP. Taken together, our results (1) do not show any transient reductive oxidative stress under hypoxia, (2) suggest that mitochondrial H2O2 production is independent of respiratory activity in the OXPHOS or LEAK state under intracellular oxygen conditions, and thus (3) challenge the LEAK-OXPHOS- ROS paradigm, emphasizing the importance of controlling dissolved oxygen in the range of physiological intracellular conditions and avoiding artificially high oxygen levels in studies of ROS production. 5 Med & Health Dec 2018; 13(2) (Suppl): 1-351 https://doi.org/10.17576/MH.2018.s1302 SYMPOSIUM 1 Sequencing the Malaysian Human Genome MOHD ZAKI SALLEH Faculty of Medicine, Universiti Teknologi MARA, Malaysia (Email: [email protected]) Systematic investigation of human variation using genome sequencing approach is fundamental for the investigation of genetic risks towards diseases and how natural selection shapes the genetic architecture. Despite on-going research efforts, most diseases risks remained un-elucidated. In addition, these efforts covered populations from the major continents; population of Southeast Asia which is known to be genetically unique has not been systematically studied. We thus aimed to provide an in depth characterization of genome variations of the unique Orang Asli and the Malays. The outcomes of these studies will provide platforms for the development of models and algorithms for disease prediction and prevention, changing the paradigm of current medical practices via precision health. We have the largest Orang Asli whole genome database known to date. All the genomes were assembled and the variants were annotated using GATK Best Practice workflow, SIFT and Polyphen. The Orang Asli genomics architectures, SNPs associated with diseases and novel SNPs were identified. The Orang Asli have a
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