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Qatar Foundation Annual Research Conference Proceedings Volume 2018 Issue 2
- Conference date: 19-20 Mar 2018
- Location: Qatar National Convention Center (QNCC), Doha, Qatar
- Volume number: 2018
- Published: 15 March 2018
41 - 60 of 82 results
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Qatar Steps up to Global Health Security: A Reflection on the Joint External Evaluation 2016
Authors: Mohamed Osman Bala, Mohamad Chehab and Nagah Abdel Aziz SelimSince the commencement of the International Health Regulations (IHR, 2005) in 2007, global public health security has been faced with numerous emerging and ongoing events. Moreover, the Joint External Evaluation (JEE) is a voluntary tool developed in compliance with the Global Health Security Agenda (GHSA) that represents a reaction by the international health community towards the increased incidence of emerging and re-emerging diseases. Against this background, between 29th May and 2nd June 2016, a team of WHO consultants arrived to the State of Qatar to assess, in collaboration with national experts, the country's capacity to prevent, detect, and rapidly respond to threats of public health aspect. They identified areas of strength, weakness, and recommendations for improving national health security of Qatar in anticipation of the 2022 World Cup event. Qatar has demonstrated a leading role in the region through its commitment to International Health Regulations (2005) and community. Similarly, the Qatar was the first Arab state and seventh volunteering country globally to undergo the JEE process. In this review, we highlighted Qatar's achievements and shortcomings of IHR core capacities to inform healthcare professionals and the scientific community about the country's contribution toward global health security.
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Bioinformatics and Virtual Metabolomics Core infrastructure to support omics research in Qatar
Authors: Anna Halama, Shaza Zaghlool, Aditya Bhagwat, Michal Kulinski, Aishah Latiff and Karsten SuhreMetabolic profiling assembles the information on the small molecule (metabolites) composition in the body. There are several factors like genetic makeup, protein abundance, environment, life habits or gut microbiota, which all contribute to “metabolic fingerprint” - specific for each individual. Metabolomics can be deployed to draw a map of metabolic interactions associated with diseased condition; characteristic metabolic alterations can be used to monitor disease onset, progression or response to the treatment. For instance, alteration in branch chain amino acids, aromatic amino acids and lipids was associated with prediction of type 2 diabetes (T2D). Furthermore, elevated level of 1,5-anhydroglucitol (1,5-AG) in plasma and saliva samples was determined by us and by others as biomarker, complementary to Hba1C, for detection and monitoring of T2D. Metabolomics study requires expertise in experimental design, sample measurements and data analysis to produce meaningful, reliable and reproducible data. Metabolomics data can be further integrated with other “omics” including transcriptomics, proteomics, glycomics and methylomics. Here we are presenting the “Bioinformatics and Virtual Metabolomics Core”, an infrastructure, which we established in Qatar to enable metabolic profiling as well as analysis of highly complex “omics” data. To perform metabolomics study the following components are required: 1) Technology providing reliable data with high quality; 2) Pipeline for data processing; 3) Expertise in data interpretation. 1) We were actively involved in establishment of targeted and non-targeted metabolomics platforms in Qatar. Targeted platform: In collaboration with Translational Research Institute (TRI) we established targeted metabolomics platform able to quantify 163 metabolites including amino acids, carnitines, phosphatidylcholines, lysophospholipids and sphingomyelins. This platform is deploying Absolute IDQ Biocrates Kit technology for sample preparation and processing. The measurements are performed using flow injection analysis (FIA) - mass spectrometry (MS) technology based on combined ExigentEksperLC100 and QTRAP4500 (ABSCIEX). Evaluation of the mass spectrometry data is performed with MetIDQ software package (integral part of the Absolute IDQ assay). Untargeted platform: We were involved in the process of untargeted platform establishment. This platform is hosted and operated by Anti-Doping Lab Qatar (ADLQ) and enables for semi quantitative profiling of metabolites in human biofluids, tissue samples and cell culture. The protocols for sample processing and preparation were optimized by Metabolon and are conducted in Qatar accordingly. These measurements are deploying Waters ACQUITY ultra-performance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer. Compounds identification is performed by comparison of peaks to library entries of purified standards based on retention index (RI). 2) The metabolomics study generates large amount of highly complex data, which requires further data processing. We have developed an internal pipeline for data processing, including quality control and data normalization. For visualization purposes we use Principal Component Analysis (PCA) and Orthogonal Projections to Latent Structures (OPLS). Statistical data analysis is tailored according to the experimental design and data structure. We are using web-tools to enable our customer direct data access. We also developed a tool called “AUTONOMICS” for automated data analysis, which can serve more advanced users. 3) Our team is extensively working with metabolomics data; we are using Gaussian graphical modeling to reconstruct metabolic pathways and to draw network of interactions between metabolites. For pathway analysis and data interpretation we are using KEGG database, Ingenuity® Pathway Analysis (IPA®) and our experience. To provide detailed overview on human disease including diabetes, kidney and liver disorders as well as cancer we are combining metabolomics with other “omics” data. Our expertise lies in processing and evaluation of data from human biofluids (urine, plasma, saliva). The team is also specialized in metabolomics in cell culture; we developed a pipeline for cell culture-optimized sample collection and processing. In conclusion, our platform was established to enable fully comprehensive metabolomics study in Qatar, which will serve local scientific community. We are supporting our collaborators and users over the whole process related to metabolomics study including experimental design, sample processing, metabolite measurements and data analysis.
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Adipose Tissue Derived-Mesenchymal Stromal Cell: Setting the ground for Clinical Grade Production at Sidra's GMP Facility
Adipose Tissue Derived-Mesenchymal Stromal Cell: Setting the ground for Clinical Grade Production at Sidra's GMP Facility1Asma Al-Sulaiti, 1 Moza Al- Khulaifi, 1Sara Al-Khawaga, 1Zohreh Tatari Calderone, 1Bella S. Guerrouahen, 1Chiara Cugno. 1 Clinical Research Center, Research Division, Translational Medicine, Sidra Medicine, Qatar. Correspondence: Dr. Chiara Cugno, MD, Director Clinical Research Center, Research Division, Translational Medicine Sidra Medicine, Qatar. Keyword list: MSC, Adipose tissue, SVF, GMP, ISCT criteria. Word count: 702Background: Among adult stem cells, mesenchymal stromal cells (MSCs) represent a highly examined population given their exceptional biological properties. The International Society for Cellular Therapy (ISCT) define human MSCs based on the following criteria: adherence to plastic in standard culture condition, expression of the surface molecules CD105, CD90, and CD73, and lack of expression for CD34, CD45, HLA-DR ( < 2%), CD14 or CD11b, CD79a or CD19, and finally capability to differentiate in vitro into osteocytes, chondrocytes, and adipocytes. The US National Institutes of Health database has reported 493 MSC-based clinical trials as of June 2015. Most clinical trials were implemented towards evaluating the biomedical potential of MSCs in treating graft versus host disease, hematological and inflammatory diseases diseases, organ transplantation, diabetes, and diseases in the liver, kidneys, and lungs, cardiovascular, bone and cartilage, neurological, and autoimmune disorders. Although multipotent MSCs are more commonly isolated from bone marrow (BM), adipose tissue-derived MSCs (AT-MSCs) represent a valid alternative source for MSCs due to their greater abundance, simple accessibility, and better immunomodulatory properties. Aim: We ultimately aim to develop the clinical-grade production of human AT-MSCs for Cellular Therapy at Sidra Good Manufacturig Practise (GMP) Facility. Our study represent the stepping stone for developing an MSC bank of high quality, reliable and well-characterized cells timely available for potential future clinical applications, such as Type 1 Diabetes (T1D). Objective 1 focuses on the isolation, expansion and characterisation of AT-MSCs. Objective 2 focuses on optimizing AT-MSCs production protocols to comply with the requirements of producing clinical-grade AT-MSCs. Methods: A) Sample collection: Adipose tissue is collected and obtained as waste material after liposuction procedures carried at Plastic Surgicenter, Doha. Aspirated fat is transferred to the Clinical Research Center at Sidra Medicine in sterile containers. B) Isolation and expansion of AT-MSCs: Both fat and blood/saline portion of lipoaspirate are used. AT-MSCs are isolated from the fatty portion, using a collagenase-based enzymatic method and from the blood / saline portion, using a non-enzymatic digestion. The isolated AT-MSCs are seeded and cultured in 75 cm2 flasks for expansion till passage 5. C) Phenotypic characterization, multilineage differentiation capacity and clonogenic assay: After reaching more than 95% confluence in the flasks at passage 1 (around 14 days in culture), phenotypic characterisation of MSCs is verified using a multi-color flow cytometry for the following specific antibodies: CD45, CD34, CD14, CD19, HLA-DR, CD105, CD73 and CD90. The adipogenesis and osteogenesis differentiation potential of AT-MSCs is also assessed at low passage. AT-MSCs are cultured under differentiating conditions in separate culture vessels. Cultures are processed for Oil Red O staining to evaluate adipogenesis differentiation (from day 7 to 14). Osteogenesis differentiation can be visualized after 21 days, using an Alizarin Red staining. To measure the clonogenic ability of AT-MSCs, Colony Forming Unit (CFU) is carried out using crystal violet staining. D) Translating AT-MSCs production to GMP facility: all protocols are optimized and standardized to comply the requirement of producing clinical-grade MSCs at Sidra Medicine GMP facility. The isolators (Biospherix XVIVO System) provides a strictly controlled environment that assures manufacturing of sterile, potent and uncontaminated products. It consists of modular sets of closed incubators and closed hoods, all integrated together as co-chambers and sub-chambers. The system is completely closed, with aseptic conditions throughout, and advanced controls as for GMP international standards. Results: AT-MSCs displayed a fibroblast-like morphology. The ISCT-recommended specific cell surface signature was verified: AT-MSCs were positive for CD105, CD90 and CD73 and negative for CD45, CD34, CD14, CD19 and HLA-DR. The biochemical assessment of adipogenic differentiation showed induced adipocytes with vacuoles containing fatty acids positive for Oil Red O staining. Alizarin Red staining showed induced osteocytes with calcium phosphate deposits. The stemness property of AT-MSCs was also confirmed. Conclusion: Recent studies have indicated MSCs as a powerful tool for several clinical applications. We are developing AT-MSCs clinical grade production at Sidra GMP Facility. We established standard operating procedures following the highest GMP international standards. Sidra GMP Facility is the first of its type in Qatar for cell therapy purposes, bringing future perspectives for the establishment of an AT-MSC bank from third-party healthy donors, after adequate screening for allogeneic donation, and a clinical trial in T1D to tackle the unmet need of preventative studies in this area.
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Prevalence of Impaired fasting glucose in Qatari women between aged of 18 to 40 years old from the Qatari Bio Bank
Authors: Youssra Dakroury, Abeer Elshewehy, Soha Dargham, Eric Kilpatrick, Lina Ahmed and Stephen AtkinBackground Impaired fasting glucose (IFG), prediabetes, has a conversion to diabetes of 10% yearly, contributing to the prevalence of diabetes of 16.7% in Qatar. This high prevalence of diabetes will translate into serious vascular complications with considerable individual early mortality.Methods The Qatar Biobank is an ambitious national project that is collecting the demographic and biological data on an unselected Qatari population that currently has data from 1100 women aged 18-84 years. Data extraction of all women between the aged 18 to 40 years (inclusive) was undertaken by the Qatar Bio bank that identified 750 women in total. The number of women with and without impaired fasting glucose (IFG: plasma glucose 5.5-6.9 mmol/l; ADA diagnostic criteria) was determined and their demographic details compared. Bio statistical analysis was undertaken using SPSS. Results The prevalence of IFG was found to be 8.3% (62/750 women). Patients with IFG had a higher cardiovascular risk profile with a higher waist hip ratio (P < 0.01), lower HDL cholesterol (p < 0.01), higher insulin levels (( < 0.01), higher pulse wave velocity (a marker of arterial stiffness, p < 0.01), higher systolic and diastolic blood pressure (p < 0.01 and p < 0.03, respectively). C reactive protein, a marker of inflammation was also elevated (p < 0.04).Conclusion Whilst this is a small cohort, 8.3% of women had a ADA diagnosis of IFG with an adverse cardiovascular risk parameter profile. This study highlights that there is a need for early screening for IFG and intervention for the prevention of diabetes in young Qatari women below the age of 40. Acknowledgments We are grateful to the Qatar Foundation and the Qatar Biobank for their support and assistance.
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CYP2C19 Genetic Polymorphism Prevalence in Qataris
Authors: Hazem Elewa, Zainab Omer Ali and Loulia BaderBackground Clopidogrel is used by around 40 million patients worldwide to treat and prevent thrombotic events. Third of the patients have clopidogrel resistance which may lead to treatment failure. cytochrome P450 (CYP450) 2C19 enzyme (CYP2C19) is one of the key enzymes involved in the hepatic bio-activation of clopidogrel. Several studies have shown that polymorphisms in the gene encoding the CYP2C19 contribute to variability in response to clopidogrel. There is no available data on the distribution of CYP2C19 genetic polymorphisms in the Qatari population. Objective The objective of this study was to estimate the distribution of CYP2C19 genetic polymorphisms in Qatari population by determining the prevalence of CYP2C19*2, *3 and *17 alleles. Methods The study was conducted on a cohort of 129 adult Qatari individuals. DNA was extracted mainly from saliva samples by Oragene® self-collection kit and purified using the prepIT™√L2P purification protocol (DNA Genotek, Inc., Canada). For those who were unable to provide saliva, blood sample was collected. Five milliliters of peripheral blood were collected in test tubes containing EDTA and DNA was extracted using PureLink® genomic DNA extraction kit for purification of genomic DNA (Invitrogen life technologies, Germany). The amount and purity of the DNA samples were quantified using Nanodrop 2000 Spectrophotometer (Thermo Scientific, Wilmington, DE, USA). Genotyping was performed for CYP2C19*2 (rs4244285), CYP2C19*3 (rs4986893), and CYP2C19*17 (rs12248560) using Fast 7500 Real-Time PCR System (Applied Biosystems™, USA) with TaqMan assays according to the manufacturer's instructions. Genotype frequencies were tested for deviations from Hardy–Weinberg equilibrium through chi-square analysis at P value of 0.05. Patients were categorized according to their CYP2C19 genotype into: ultra-rapid (*1/*17, *17/*17); extensive (*1/*1); intermediate (*1/*2, *1/*3, *2/*17); poor (*2/*2, *2/*3, *3/*3) metabolizers. Results This study included 129 Qatari individuals. Majority of our patients were 50 years and above (60%) and mostly females (63%). The allele frequencies were CYP2C19*1 (86 %), *2 (4%), *3 (0%) and *17 (10%). Fifty-eight subjects (45%) were homozygous for the *1/*1 genotype. Heterozygous genotypes were identified in 62 patients (48.1%), 42 carrying *1/*17 (32.6%), 12 carrying CYP2C19 *1/*2 (9.3%), and 8 carrying CYP2C19 *2/*17 (6.2%). Six patients were homozygous for the gain-of-function allele, CYP2C19 *17/*17 (4.7%). On the other hand, three patients were homozygous for the loss-of-function allele, CYP2C19 *2/*2 (2.3%). However, no subject was found to have *3 allele. All the genotypes were in Hardy–Weinberg equilibrium. The distribution of CYP2C19 phenotypes was divided into extensive metabolizers (*1/*1) (45%), ultra-rapid metabolizers (*1/*17, *17/*17) (37.2%), intermediate metabolizers (*1/*2, *1/*3, *2/*17) (15.5%), and poor metabolizers (*2/*2) (2.3%). Conclusion To the best of our knowledge, this is the first study to report the distribution of CP2C19 genetic polymorphisms in Qatari population. These findings have important clinical implications for the use of clopidogrel and other medications affected by CYP2C19 mutations in Qatari population. Thus, future association studies are needed to reveal clinical consequence of these genetic polymorphisms in Qataris.
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Combination novel modalities for enlightening global molecular elemental ultrastructure and morphological biochanges in subacute stroke in rat brain
By Mohamed AliBackground and Purpose: Stroke is the main cause of adult disability in the world, leaving more than half of the patients dependent on daily assistance. Novel and advanced diagnostic modalities are required to improve treatment in order to reduce the social and economic burden of this disease. Understanding the post-stroke bio-chemical changes is critical for patient treatment and survival. Unfortunately, conventional diagnostic procedures are used to investigate global changes in the stroke brain, rather than understanding the bio-chemical changes at the molecular and cellular level. Therefore, in the current study we used combined approaches to investigate the molecular, elemental, ultrastructure and morphological changes in stroke brain. Methods: Application of Fourier transform infrared (FT-IR) spectroscopy; laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS); transmission electron microscope (TEM), Raman micro-spectroscopy and atomic force microscopy (AFM) were used to examine stroke brain and were compared with the standard approach of immunohistochemistry. The areas investigated, one-week post sub-acute photo-thrombotic ischemic stroke in male rats, were primary stroke lesion gray matter (PS-GM), perilesional gray matter (PL-GM), lesioned white matter (L-WM), contra-lesioned white matter (CL-WM), contra-lesioned gray matter (CL-GM) corpus callosum (CC) and dorso-ventral cortices (DVC). Results: The infrared spectroscopy results revealed that total lipid and protein and the lipid CH2 content decreased significantly in the (PS-GM) while the total aggregated protein increased significantly relative to total protein, in particular we detected an increase in Aβ1-42. In contrast, both (PL-GM) and (L-WM) regions experienced an increase in carbonyl esters, olefinic = CH, and the CH3 groups of lipids. In contrast, both (PL-GM) and (L-WM) regions experienced an increase of carbonyl esters, olefinic = CH, and CH3 groups of lipids which is associated with decrease in CH2 lipid groups and lipid contents. Amongst the novel findings of the study was that the aggregated protein detected in the L-WM was Aβ1-42. FT-Raman micro-spectroscopy showed a reduction in lipid content and iron accumulation around the stroke lesion in the form of heme. Elemental analyses for divalent cations: Ca, Cu, Fe and Zn realized major changes in the different brain structures that may underscore functionality within these regions. In the elemental maps high Fe intensity around the stroke region correlated well with the presence of heme shown by Raman spectroscopy. Conclusions: The complementary methodologies revealed, at multiple-levels, the effects of stroke: edema, lipid peroxidation, as detected by increased olefinic = CH and carbonyl ester content, and enhanced protein aggregation. For the latter, an example was the identification of Aβ1-42 in the L-WM. These data point to the possibility that amyloid plaque formation is not exclusive to cortical neurons, but also affects the WM. One conclusion being that application of strategies developed for AD-treatment may be fruitful in treating stroke. Alternative therapies would include targeting edema, antagonizing lipid peroxidation for the protection of the blood brain barrier (BBB) post-stroke by the application of antioxidants. Alternatively application of free Fe sequestering molecules, as FT-Raman micro-spectroscopy detected the reduction in the lipid content and the iron accumulation around the stroke lesion.
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MitochondrialDerived Peptide MOTSc promotes hepatic fatty acid metabolism and regulation by metformin
More LessMitochondrial-Derived Peptide MOTS-c promotes hepatic fatty acid metabolism and regulation by metformin. Ramanjaneya M1, Jerobin J1, Bettahi I1, Sivaraman SK1, ¬ Mohammad RM1, Skarulis MC1 and Abou-Samra AB1. 1Translational Research Institute, Academic Health System and Department of Medicine, Hamad Medical Corporation, Doha, Qatar. Objectives: Mitochondrial dysfunction contributes to the pathogenesis of metabolic disorders and is a leading cause for development of insulin resistance. This is promoted, at least partly, through lipid accumulation in ectopic tissues, including liver (non-alcoholic fatty liver disease) and skeletal muscle. The role of mitochondria as functional organelles, and the signaling molecules produced by them are critical for cellular energy homeostasis. Dysfunction in mitochondria contributes to the pathogenesis of metabolic disorders. MOTS-c (mitochondrial open reading frame of the 12S rRNA type-c) encoding a 16-amino-acid peptide is a recently identified peptide. The skeletal muscle appears to be the main target organ for MOTS-c and its cellular actions inhibit the folate cycle and its tethered de novo purine biosynthesis, leading to AMPK activation. MOTS-c promotes glucose utilization, insulin sensitivity and metabolic homeostasis through activation of AMPK dependent mechanisms in skeletal muscle. MOTS-c protects rodents against diet and ageing induced insulin resistance. However the role of MOTS-c hepatocytes is unknown. In this current study we aimed to study the functional importance of MOTS-c on liver fatty acid metabolism in using HepG2 hepatocytes model. Methods: Analysis of MOTS-c gene expression in human tissue panel: Human tissue cDNA panel was obtained from Clontech. MOTS-c gene expression in various human tissues was measured by real-time PCR. Human hepatocytes (HepG2) cells were used for this study: Cells were maintained as subconfluent monolayers in DMEM (Invitrogen) with 10% fetal bovine serum and 100 units/ml penicillin plus 100 μg/ml streptomycin (Invitrogen) at 37 °C with 5% CO2. For all stimulation experiments cells were grown for overnight in 2% serum containing media. Fatty acid-induced steatosis: Approximately 80% confluence were pretreated with MOTS-c and further stimulated for 24 hrs with oleic acid and palmitic acid 1:2 ratio in 2%-serum medium for 24 hrs in 12-well culture plate. Following this incubated with nile red solution for 20 mins and then subjected to flow cytometric analysis to measure lipid accumulation in HepG2 cells. Control cells were treated with fatty acid-free medium containing albumin. Effects of antidiabetic drugs on MOTS-c expression: Approximately about 80% confluent cells were stimulated with commonly used antidiabetic drugs insulin 100 nM, rosiglitazone 2 mM or metformin 1 mM for 24 in 2%-serum medium 12-well culture plate. Following this MOTS-c secretion was measured using commercially available elisa kits. Results: MOTS-c gene transcripts are expressed in human liver and HepG2 cells. Stimulation of HepG2 cells with MOTS-c suppresses fatty acid synthase (FAS), sterol regulatory element-binding protein-1c (SREBP1c) and acetyl-CoA carboxylase (ACC) the key genes required for hepatic lipid production. Further MOTS-c reduced fatty acid induced hepatic lipid accumulation. Conclusion: Our primary result indicates that MOTS-c promotes hepatic fatty acid metabolism through down-regulation of multiple genes involved in fatty acid metabolism in liver. Further studies are in progress to assess the effects of anti-diabetic drugs such as metformin, insulin and rosiglitazone on MOTS-c secretion. The findings from this study are of clinical importance given that non-alcoholic liver disease (NAFLD) is highly prevalent in obese and diabetes subjects in Qatar. MOTS-c and its agonists have potential to be used as therapeutic agents for treatment of NAFLD.
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Foreskin DerivedMesenchymal Stromal Cell FSKMSC: Setting the ground for the Clinical Grade Production of MSC at Sidra's GMP Facility
Moza Al- Khulaifi1, Asma Al-Sulaiti1, Bella S. Guerrouahen1, Sara Al-Khawaga1, Chiara Cugno1, Zohreh Tatari Calderone1 1 Clinical Research Center, Research Division, Translational Medicine, Sidra Medicine, Qatar. Correspondence: Dr. Zohreh Tatari Calderone; Clinical Research Center, Research Division, Translational Medicine Sidra Medicine, Qatar. Email: [email protected] Keyword list: MSC, Foreskin, GMP, ISCT criteria. Abstract 1. Background: Mesenchymal stromal cells (MSCs) are known for their multipotent, immune-privileged and regenerative properties. MSCs can be obtained from different sources such as bone marrow, fat, umbilical cord, synovial fluid, tendons, muscles, periosteum, peripheral blood and nervous system. The International Society for Cellular Therapy (ISCT) has stated minimal characteristics for defining the MSCs as multipotent fibroblast-like cells highly adherent to plastic, positive for CD73, CD105 and CD90 but negative for CD45, CD34, CD14 or CD11b, CD19 or CD79a, and HLA-DR. The MSCs constitute a promising tool for regenerative medicine, due to their self-renewal capacity, multilineage differentiation potential, paracrine effects and immunosuppressive properties. They are extensively used in the treatment of several clinical settings including but not limited to, cardiovascular diseases, inflammatory bowel diseases, liver dysfunction, rheumatological disorders and diabetes. Bone Marrow-derived MSCs has been considered as the gold standard for cell-based therapy, but bone marrow aspiration is an invasive method that limits the access to a large number of MSC donor sources. Recently, the foreskin, which has been always retained as medical waste, has been recently shown as a source for MSCs. Sidra Medicine is performing 4000- 5000 circumcisions annually, therefore foreskin can be considered as a highly abundant and easily accessible source for the isolation of MSCs. Therefore, the aim of our study is: Specific Aim: To demonstrate that MSC isolated from foreskins (FSK-MSC) meet the high standard criteria of ISCT and to compare their phenotype, potency and multilineage potential with MSCs obtained from a classical source such as adipose tissue for the clinical application. 2. Method: Specimen Collection and MSC Isolation: Upon approval by the IRB committee, foreskins were collected after the circumcision from the Surgery Department at Sidra Medicine (age range: 1.5 -14 years old). Epidermis was manually separated from the dermis and was then subjected to enzymatic digestion by liberase. Cell suspension was cultured in T75 flasks containing low-Glucose media, complemented with 10% fetal bovine serum and 1% antibiotics for 3-5 days. After 3-5 days, the non-adherent cells were removed and fresh media was added to allow the adherent MSCs to grow. MSC Characterization:Cell Surface expression: Phenotypic characterization was determined using flow cytometry. Briefly cells were stained for the surface markers using fluorochrome conjugated antibodies for CD14, CD19, CD34, HLA-DR, CD45, CD73, CD105 and CD90 according to the ISCT criteria. Differentiation: FSK-MSCs were treated with osteogenic and adipogenic mediums in separate cultures for 7, 14 and 21 days and stained with Alizarin Red and Oil Red O staining respectively. The lineage differentiation was visualized using the inverted microscopy. Colony Forming Units (CFU): Colony formations Units measures the viable clonogenic cell numbers that is an important indication of the quality of cell preparation. The CFU assay was performed according to commonly used protocol and using crystal violet staining. 3. Results: FSK-MSC displayed a typical fibroblast-like morphology and an ISCT-compliant phenotype; FSK-MSCs were highly proliferative and had a marked clonogenic potential. FSK-MSCs displayed osteogenic and adipogenic differentiation capacities. 4. Conclusion: The isolated FSK-MSCs were shown to be compliant with ISCT criteria and can be considered as a reliable source of MSC. Their compliance allows considering the standardization of clinical grade FSK-MSCs production at Sidra Medicine GMP facility, and setting the ground for the potential establishment of a MSC biorepository.
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Effect of nonnutritive sweetener stevioside on regulation of adipogenesis and oxidative stress in adipocytes
More LessEffect of non-nutritive sweetener stevioside on regulation of adipogenesis and oxidative stress in adipocytes Jayakumar Jerobin, Manjunath Ramanjaneya, Ilham Bettahi, Siveen Kodappully Sivaraman, Ramzi M Mohammad, Monica Skarulis Young, Abdul Badi Abou Samra Interim Translational Research Institute, Qatar Metabolic Research Institute & Department of Medicine, Hamad Medical Corporation, Doha, Qatar. Introduction The metabolic syndrome which includes obesity, diabetes, polycystic ovarian syndrome and non-alcoholic fatty liver disease is a major health risk in Middle Eastern countries including Qatar. The metabolic syndrome is worsened by unhealthy diet and sedentary lifestyle. Both nutritive and nonnutritive sweeteners (NNS) are consumed by people worldwide, and the consumption of sweeteners above the recommended level contributes to metabolic syndrome. In adipocytes, artificial NNS such as saccharin and acesulfame potassium promotes adipogenesis and suppresses adipose tissue lipolysis. The consumption of artificial NNS saccharin modulates the gut microbiota and promotes glucose intolerance. Currently the artificial NNS are replaced with natural NNS stevioside in foods and beverages since it has low caloric value. Studies have shown that stevioside possess glucose uptake properties which may have a potential role in type 2 diabetes mellitus treatment. This proposed study mainly focuses on the effect of NNS stevioside on regulation of adipogenesis and oxidative stress in matured adipocytes. Methods The 3T3L1 preadipocytes cells were cultured in DMEM medium, and the preadipocyte cells were differentiated in to mature adipocytes by the addition of IBMX, Insulin and Dexamethasone on 0th day. It was replaced with maintenance medium containing insulin on 2,4 and 6th days of differentiation. The stevioside at different dosage (10, 100 and 200 μM) were treated on 0, 2, 4 and 6th days of differentiation in matured adipocytes. Cell viability was analyzed using MTS solution. Oil red¬ O staining was carried out to study the accumulation of lipids in the undifferentiated and matured adipocyte cells. The different oxidative stress markers such as Reactive oxygen species (ROS) and reduced Glutathione were measured using flow cytometry. Western blot and gene expression studies were carried out to study the adipogenesis and insulin sensitivity markers. Results The stevioside was found to be non-toxic to both preadipocyte and adipocytes. On eighth day of differentiation, a significant increase in the accumulation of lipids was observed in the differentiated adipocytes compared to the undifferentiated preadipocytes as analyzed by Oil Red O. The generation of oxidative stress was higher in adipocytes compared to the undifferentiated adipocytes. This was confirmed by increase in the level of reactive oxygen species radicals and reduced glutathione in untreated adipocytes. Treatment with stevioside resulted in inhibition of oxidative stress markers in a dose dependent manner. The protein expression level of early adipogenesis marker such as PPAR-γ and C/EBP-alpha was significantly decreased in stevioside treated adipocytes. Adiponectin, an adipokine found to have association with insulin sensitivity was significantly increased in stevioside treated adipocytes. The protein and gene expression levels of GLUT4 was also found to be significantly increased in stevioside treated matured adipocytes. Conclusion Stevioside reduces oxidative stress by modulating the level of free radicals in maturing adipocytes. Adiponectin an insulin sensitizer which was found to be lower in obese individuals was significantly upregulated in stevioside treated adipocytes. Similarly, the increase in the expression of insulin regulated glucose transporter GLUT-4 in adipocytes indicates that stevioside may possess insulin sensitivity properties in matured adipocytes.
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Generating an imagingbased approach for enhanced structural and functional analysis of zebrafish cardiovascular systems
Authors: Huseyin Yalcin, UmmaySalma Abuhabib, Naila Kitaz, Ala Mohamed and Zain ZakariaCardiovascular diseases (CVDs) have latterly become one of the leading cause of death and impairment worldwide. According to the World Health Organization (WHO), data estimation in 2008 were about 17.3, 7.3 and 6.2 million death cases for CVDs, heart attacks, and strokes, respectively [1]. Congenital heart defects (CHDs) are important forms of CVDs affecting about 1% of the population. According to Dr. Ahmad Sallehuddin (Consultant and Chief of Pediatric Cardiac Surgery at HMC), CHD incidence in Qatar is about 6-8 in every 1,000 births and about 100 new patients with CHDs require surgery every year [4]. Both genetic and environmental factors were shown to contribute to CVDs. Human genetic studies are not sufficient alone to explain the genetic basis of CVDs due to disease heterogeneity, inconsistent penetrance, and predominantly a delayed onset of symptoms. Therefore, animal models are necessary to investigate and distinguish novel genes that contribute to the pathology of CVDs and also to unravel environmental factors that play role [5]. More recently, the Danio rerio (Zebrafish) has emerged to become an intriguing vertebral model in medical science research. This model has several advantages, such as an almost entirely sequenced genome that is highly preserved with humans: approximately 70% of the human genes are estimated to have orthologue genes in the zebrafish genome [2,8]. The features of the zebrafish over mammals as a vertebral model include its simplicity of genetic manipulation, a large quantity of offspring, and their external and fast development [5,8]. Transparency of zebrafish embryos provide precise observation of the heart beats, heart chambers and circulating blood in vivo via light microscopy [5]. Additionally, passive diffusion is sufficient for oxygen delivery during early stages of zebrafish embryos development because they are independent of circulatory system [7]. Therefore, embryos having intensive cardiac faults survive early development, which enables investigation of mutations whereas mammalian models are not appropriate. There are many genetic cardiovascular models for zebrafish that allow researchers to investigate cardiovascular diseases. Novel techniques are needed for these models to assess comprehensive and quantitative phenotyping of mutant hearts and blood vessels. These analyses involve determination of the cardiac atrial and ventricular shortening fractions, examination of the blood flow velocities and recording of the electrocardiograms. In addition to genetic studies, cardiotoxicity investigations for drugs or teratogens necessitate heart function and morphology assessment. For the current study, the aim is to adapt previously established experimental and image analysis techniques to zebrafish studies, which will enhance structural and functional analysis of the zebrafish cardiovascular systems in biomedical research Methodology: This project involves generation of two main protocols, which are for structural and functional analysis of zebrafish embryos. Both these analyses are required to be performed to assess the severity of induced heart defects in zebrafish embryos. For the structural analysis, we will perfuse zebrafish (3 -5 days post fertilization (dpf)) hearts with microfilm, a CT- dense agent. Once it polymerises, a cast is generated for heart lumen. The embryos are then fixed with paraformaldehyde and scanned via micro-CT. This procedure provides 3D images for the cardiovascular systems of the animals enabling measurement of the volumes of the heart chambers. This technique enables us to measure the effect of induced heart mutations on the heart morphology during the embryonic growth of the animal. We have previously used this technique for visualization of cardiac chambers for embryonic chicken successfully [3,10]. The approach is adapted for zebrafish studies in the current work. For the functional analysis, image sequences of the beating ventricles and the red blood cell (RBC) movements of the zebrafish larvae (3 -5 dpf) are recorded at a camera speed of 70 fps and above. This imaging is done by taking videos using Micromanager Program that is connected to a stereomicroscope. To take image sequences, zebrafish larvae are placed on its lateral side, in which the right side will be on top. A variety of measurements and calculations are performed using Image J Program to assess heart functions. The parameters that are calculated include myocardial wall velocities for assessment of severity of induced heart muscle defects, and measurement of RBC blood flow velocities for assessment of rhythmic beating defects of the heart. Other parameters are fractional area change, fractional shortening, heart rate, stroke volume, cardiac output and ejection fraction [9]. Stroke volume, is the blood volume that is ejected from the heart in each beat, cardiac output, is the amount of circulating blood across the heart in each minute, and ejection fraction, is the measurement of blood that is ejected from the ventricle with each beat. This will provide more precise and accurate evaluation on the heart function for genetically mutated animals to be compared with the normal ones. Several zebrafish mutants display phenotypic features resembling human cardiac diseases. These mutants whose molecular damages have been determined like mutations in regulatory myosin light chain, titin, cardiac troponin T, essential myosin light chain, and beta-myosin heavy chain, have been linked with cardiomyopathy in humans [6]. Therefore, reduced contractility, myocardial wall velocities, and cardiac output can be studied as a sign of having cardiomyopathy. Accordingly, opening ways to apply drugs for morphological and structural evaluation of the heart. In conclusion, in this study, we are developing imaging-based techniques for accurate structural and functional analysis of the zebrafish hearts through adapting previously established methods on another animal systems. Once we establish our approach, analysis methods will be readily available for other Zebrafish researchers.
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The triple interaction DietMicrobiomeEpigenome: new approach to the noncommunicable diseases
Background/Objective: Qatar suffers the highest percentage of non-communicable diseases (NCD), such as diabetes and obesity among few countries in the world. There are two major components that could explain the NCD, which are diet and the genetic background. It's well demonstrated that diet affects the genome function in terms of epigenetic and microbiome modification. Recent scientific advancement or studies have shown that all these components could be the factors in defining the development of the NCD, but, there is no study that explains the direct connection of Diet-Microbiome-Epigenome. We want to explore the interaction of these three factors in a small pilot study on type 1 diabetes (T1D) creating a methodology that will be applicable for all NCDs and nutrition-related diseases. This study aims to identify specific nutrients that modulate gut microbiome; to define different microbiome composition and metabolites; and to define differential methylated regions (DMR) that is possibly affected by nutrients, gut microbiome and its metabolite. This abstract presents the study design and discusses methodologies that will be used in the study. Study design and methodology: We will compare 4 groups of subjects: T1D, T1D obese, pure obese, and lean controls. Pediatric patients will be recruited from Sidra OPC based on major inclusion criteria, (such as age should be between 6-12 yrs, no antibiotic treatment in the past 3 months, no chronic diseases except T1D and no history of cancer) and divided into four groups: (1) healthy lean children (5-84th percentile of BMI), (2) Obese ( ≥ 95th percentile of BMI;), (3) T1D and (4) Obese T1D. A comprehensive set of physical measurements (body weight, height and Waist circumference), clinical biomarkers for diabetes and obesity (mainly, blood glucose level, lipid and liver profile, HbA1c will be done with blood sample) and family history of diabetes, treatment history will be collected and most importantly dietary habits will be collected by 24 hrs food recall. Two sets of stool samples (one for microbiome analysis by 16S rDNA-sequencing; and one for fatty acids analysis by gas chromatography) and blood samples (one for DNA extraction for methylation analysis by Illumina DNA-methylation Array and one for RNA extraction for gene expression analysis using Fluidigm platform) will be collected from each subject. Data analysis will investigate any association of diet to the clinical phenotypes comparing the four groups of subjects, using logistic regression; two-sided P-value of < 0.05 will be considered statistically significant. Possible Research Outcomes/Conclusion: At the end of pilot study, we can able to define the (1) methodology workflow, (2) nutrients list or diet patterns that increase the risk of T1D in obese children, (3) specific microbiome pattern, in terms of composition and metabolite, in obese T1D children, and (4) specific nutrients and microbiome metabolites that alter DNA-methylation and gene expression in obese T1D children. This methodology workflow will be applied to other studies on NCD and nutrition-related diseases.
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Healthcare practitioners' views of their role in addressing the medical comorbidites of people with mental illness
More LessBackground: The lifespan of individuals with serious mental illness (SMI) is shorter compared to the general population. These disorders include schizophrenia, schizophrenia-like psychosis (e.g., schizoaffective disorder), major depressive disorder (MDD), and bipolar affective disorder (BPAD). This excess mortality is mainly due to physical illness and the lack of preventive medical care that is provided for this sector of the population. Although this has been explored Objectives: To investigate the prevalence rates of different physical illnesses in individuals with SMI in Qatar and to examine how these are being managed. Also, to explore health care practitioners' (HCPs) role, awareness and views on addressing the medical comorbidities of people with mental illness. Methodology: The published literature was explored by searching various electronic databases (PubMed®, Embace®, CINAHL®, PsychInfo®) on prevalence rates, morbidity and excess mortality rates in SMI. This was followed by a cross-sectional retrospective chart review of a cohort of patients with SMI attending the outpatient psychiatric clinic at Hamad Medical Corporation (HMC). A comprehensive electronic data extraction tool using SurveyMonkey® was used to collect patient demographics, psychiatric and medical co-morbidities, psychiatric and non-psychiatric medications, monitoring laboratory parameters and all relevant physical assessment findings such as blood pressure, weight and height. SPSS® was used for data analysis. The final phase consisted of semi-structured interviews with HCPs working in different health sectors (hospitals and primary health care centers). Thematic analysis was used to explore themes related to their views and perceived roles in addressing the medical comorbidities of people with mental illness. Results: The literature review yielded 792 relevant citations of which 17 met the inclusion criteria. Compared to the general population, metabolic, cardiovascular, respiratory, and musculoskeletal diseases were found to be more prevalent among severe mental illness patients from a global perspective. Of three hundred thirty six patients with SMI who were eligible for the retrospective chart review, almost a one third (29.2%) had at least one medical comorbidity documented. Diabetes was the most frequent, diagnosed in (16.1%) of these patients, followed by dyslipidemia (9.8%) and hypertension (9.2%). Monitoring of the risk factors associated with the comorbidities and other relevant physical assessment parameters (such as blood pressure, weight, HbA1c, blood glucose and lipids) were documented for less than 50% of patients, and some parameters, such as smoking status, were not documented at all. A total of eighteen face-to-face interviews with HCPS were conducted in the second phase of the study from which four major themes emerged, including 1) knowledge and awareness, 2) perceptions of current practices, 3) perceived barriers to care and 4) solutions to overcome these barriers. Conclusion: Results from the two exploratory phases of this study suggest that individuals with SMI in Qatar are less likely to receive standard levels of care for their medical comorbidities and less likely to be followed-up regularly. Poor documentation and lack of adherence to key practice guidelines for the provision of a holistic approach to care for individuals with mental illness may be contributing to fragmentation of care which will need to be addressed.
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Antidiabetic and Antioxydant Activities of Various Extracts From Different parts of Cleome Arabica Grown in Algeria
Authors: Fatiha Seglab, Yousfi Mohamed and Ihcen KhchebaBackground Diabetes mellitus (DM) is a chronic metabolic disorder caused by an absolute or relative lack of resistance to insulin. It is characterized by hyperglycemia and accompanied by various chronic vascular complications. One therapeutic approach for managing blood glucose-and thereby preventing or delaying the above-mentioned complications-is to decrease the catalytic activity of key enzymes involved in hydrolytic cleavage of dietary oligosaccharides such as α – amylase and α – glucosidase. Plants are used from antiquity as sources of medicament against various diseases. These properties are usually attributed to secondary metabolites which are the subject of a lot of research in this field. in particular phenolic compounds (class of naturally occurring pigments with ubiquitous distribution in plant Kingdom) which they display a remarkable biological properties [3]. Many plants are drug-design targets for the development of compounds for treatment of diabetes [1] Therefore, safer natural α - amylase inhibitors have been reported from plant sources [2] The Algerian flora, is one of the richest in the world, with its many species belonging to several botanical families of which a large percentage endemic, remains very little phytochemically as pharmacologically explored. In keeping with the general pattern of bringing one's contribution to the development of the vegetable reign as a source of natural bioactive substances we will be interested in the ethnobotanical and phytochemical study of some Algerian plants to discover a new therapeutics compounds, The expertise of our laboratory in natural substance from medicinal plants obtained by extractive phytochemistry reported the screening results for α - amylase inhibitory activity of more than 30 herbal extracts. This plant species were submitted to chemical screening, the analysis of the preceding biological targets led to the evaluation of the biological activity of the extracts of the specie Cleome arabica, to confirm and distinguish their antidiabetic activity by the inhibition of α-amylase.[4][2] A B S T R A C T The aim of this study consisted on the investigation of the antidiabetic and antioxidant activities of various extracts from Cleome Arabica collected on two seasons in the town of Laghouat in the steppe region of Algeria. The preliminary evaluation of the phytochemical composition of this extracts highlighted the presence of some chemical groups. This was confirmed by a quantitative analysis based on the determination of phenolic, flavonoid and condensed tannin content. The results of the effects of phenolic compounds on the kinetic catalyzed by the α – amylase using an in vitro model, to find a natural anti- diabetic compound from the plant indicate that the phenolic extracts from these plant have an inhibitory effect on the enzyme. The antioxidant activity test shows that our phenolic extracts exhibit good antioxidant capacity. Our study is the first report on potential inhibition of these plant extracts of digestive enzyme
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Harnessing Qatar Biobank to Understand Type 2 Diabetes in Adult Qataris
Authors: Ehsan Ullah, Raghvendra Mall and Halima BensmailObjectives: Chronic diseases such as diabetes, obesity and cancer are caused by the complex interaction between environmental factors (such as diet, lifestyle, and the built environment) and genetic factors. To understand the ultimate role of environmental, behavioral, and genetic factors along with their interactions, large-scale population cohorts have been established, mainly in Europe, North America, China, Japan, and Korea. The Qatar Biobank is a Qatar national population based prospective cohort study which includes the collection of biological samples, with long-term storage of data and samples for future research. The ultimate goal is to allow physicians and researchers to use the data collected from the biobank to conduct a large-scale study of the combined effects of genes, environment, and lifestyle on these diseases, to educate people on risk factors for these common diseases and to study disease incidence patterns and develop new diagnostic and therapeutic approaches. Using this pilot data, we had access to 60 features measured on 1000 Qatari citizen. To the best of our knowledge, this is the first study that has been done on Qatari biobank few months after its release. The main objective of the study is to identify the associated risk factors in Qatari population compared to those previously found in other parts of the world. Methods: In this study, we apply a panorama of state-of-the-art statistical methods and machine learning algorithms to investigate risk factors for diabetes. The statistical methods rely on lasso and group-lasso based techniques that can even use mixed continuous and categorical variables. The machine learning methods rely on tree-based models that provide importance of variables in predictions. In contrast to relying solely on the widely used baseline statistics, which perform marginal analysis considering a single variable at a time, these methods are based on multivariate analysis of the medical conditions. Moreover, we have applied survival and risk analysis on the prognosis of diabetes in the Qatari population. In our analysis, we used survival analysis to estimate the distribution of time of diabetes development. We have used Cox proportional hazards model to investigate the effect of different variables on the risk of diabetes. Results: Our study strongly confirms known risk factors associated with diabetes in Qatari population as previously found in other population studies in different parts of the world. For diabetes, biomarkers in Qatari population (as identified by different methods) include magnesium, calcium, HDL-C, chloride, insulin, c-peptide of insulin which have been previously reported by to list a few. Our study has revealed interactions of hypomagnesemia with HDL-C, triglycerides, and free thyroxine. These findings need further investigations. The survival analysis reveals that at the age of 40, there are 15% chances of developing diabetes in Qatari population and the chances increase to 50% at the age of 63. Qatari females are slightly at more risk to diabetes than males before the age of 40 but later on males have more chances to develop diabetes. The risk analysis reveals that calcium, magnesium, hemoglobin, triglycerides, and free-triiodothrymine play a very significant role in determining risk of the disease in Qatari population. Conclusion: Our study strongly confirms known risk factors associated with diabetes and obesity in Qatari population as previously found in other population studies in different parts of the world. Moreover, interactions of hypomagnesemia with other risk factors merit further investigations.
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Characterization of human microbiota of biological samples collected from healthy adult volunteers using different DNA extracting protocols
Introduction:The human body harbors bacteria, archaea, viruses, and eukaryotic microbes that inhabit interactive interfaces exposed to the external environment. This collection of microorganisms consists of about 100 trillion of microbes called the human microbiota and the total genes encoded by these microbes collectively called as human microbiome. They are strongly associated with the development of non-communicable diseases (NCDs) in addition to the role they play in communicable infectious diseases. Changes in microbiota has been reported in various NCDs including obesity, cancer, diabetes, metabolic syndrome, inflammatory bowel disease (IBD), asthma, cardiovascular disease (CVD), kidney disease (KD) and others. Advances in DNA sequencing and bioinformatics had enabled researchers the exploration of the genetic diversity of the uncultured component of host-associated microbial communities. The aim of this work is to test different DNA extracting protocols to characterize human microbiota of biological samples collected from adult volunteers. Materials & Methods: Biological samples such as human saliva, vaginal swabs and stool were collected from well-characterized adult participants. Total DNA were extracted from Vaginal swabs (n = 5) using DNeasy Blood & Tissue Kit and Mobio power Soil protocol; Stool samples (n = 17) using MoBio Power Fecal and Qiamp fast stool kits; Saliva samples (n = 8) using QIASymphony DNA Midi kit. DNA quantity and quality was assessed by Nanodrop spectrophotometer. Integrity of the DNA was reviewed on LabChip. Human microbiota diversities of different biological samples through different extraction protocols will be determined using MiSeq-Illumina high-throughput sequencing of bacterial 16S rDNA V1-V3 fingerprint. Sequenced data will be analyzed using QIIME pipeline. Results DNA measurement revealed that DNeasy Blood & Tissue Kit gave higher DNA yield with good integrity (2-8 ng/μL) than Mobio power Soil protocol (0.6-2 ng/μL) for Vaginal swabs. Likewise, MoBio Power Fecal kit (1.8-41.5 ng/mL) gave higher DNA yield than Qiamp fast stool kit (5-21 ng/mL) for stool samples. On the other hand, DNA Qiasymphony DSP kit gave (3-241 ng/μL) for saliva. Expected Outcome: From the previous studies by other groups, it is expected that Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria are the most abundant phyla in human microbiota. The abundance of the mentioned phylum may differ with respect to body sites. Actinobacteria members will be abundant on skin, Firmicutes and Bacteroidetes will be more abundant in gut. Firmicutes, Proteobacteria, and Bacteroidetes will be the more abundant in saliva and Firmicutes in Saliva. In addition, Each DNA extraction protocol has different strategies to lyse the human microbes. We speculate that the microbial diversity of same body site can be different with different protocols. Characterization of healthy human microbiome gives a better understanding of their role in human health during diseased conditions. Especially, Human gut microbiota influence the metabolism of host and it links to several metabolic disorders such as obesity, Type 2 diabetes and Metabolic syndrome when there is an imbalance in microbiota. Exploring Vaginal microbiome can guide as to find new therapeutic targets to treat several infections and to treat complications with preterm birth to pregnant women. This work will be a promising candidate to choose the adequate protocol based on the nature of the sample, which can guide us to explore the human microbiome in right path.
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1 25 Vitamin D3 levels in Qatari diabetes patients
Lina H.M Ahmed1, Youssra Dakroury1, Soha R. Dargham1, Aishah Latif2, Stephen Atkin1, Amal Robay1, Omar M. Chidiac1, Charbel Abi Khalil1 1 Weill Cornell Medicine Qatar, 2Antidoping Laboratory Qatar. Introduction: Vitamin D deficiency is a major issue worldwide and particularly in countries in the Middle East where full coverage of the body is a normal cultural practice. Epidemiology studies have suggested that vitamin D deficiency is associated with the development of diabetes where those levels are lower than normal subjects. However, the role of vitamin D in the development of diabetes complications is unclear. We hypothesized that vitamin D3 levels (25OHD3) and its active form 1,25 dihydroxyvitaminD3 (1,25OHD3) would differ between subjects with and without diabetes due to full coverage of the body being the determining factor. Methods: 499 Qatari subjects were recruited of whom 274 (54.9%) had type 2 diabetes and 222 (44.5%) did not. We used the Mann-Whitney test to compare the levels of both vitamin D outcomes, 25OHD3 and 1,25OHD3, their data were not normally distributed. First we compared the 25OHD3 and its active form 1,25OHD3 between diabetes and non diabetes subjects. Among those who were diabetic, we then correlated the 25OHD3 and 1,25OHD3 with diabetes complications (Table). 25OHD3 and 1,25OHD3 were measured by LC-MS/MS analysis. Results: Patients with diabetes were significantly more deficient in both 25OHD3 (p-value = 0.001) and 1,25OHD3 (p-value < 0.001) than non-diabetic patients. There were gender differences in the levels of vitamin D, with males observing less vitamin D levels than females (p-value < 0.001). Higher 25OHD3 levels were associated diabetic retinopathy (p-value = 0.014) but there was no difference in other parameters (Table). 1,25OHD3 deficiency was associated with hypertension (p-value = 0.043), but no other factors. Conclusion These data show that both 25OHD3 and 1,25OHD3 levels are lower in diabetes patients and in Qatari females, with only 1,25OHD3 deficiency being associated with hypertension, whilst both 25OHD3 and 1,25OHD3 deficiency were not associated with other complications of T2DM Table Alpha_D3, median (IQR) P-value 25OHD3, median (IQR) P-value Gender Gender Male (n = 97) 0.031 (0.037) 0.009 Male (n = 121) 8.26 (11.01) 0.000 Female (n = 96) 0.022 (0.032) Female (n = 153) 4.53 (7.44) Hypertension Hypertension No (n = 76) 0.033 (0.031) 0.043 No (n = 100) 6.39 (10.39) 0.612 Yes (n = 117) 0.023 (0.033) Yes (n = 174) 6.06 (10.61) Dyslipidemia Dyslipidemia No (n = 61) 0.032 (0.036) 0.166 No (n = 75) 6.86 (10.63) 0.061 Yes (n = 132) 0.024 (0.032) Yes (n = 199) 5.56 (9.00) Diab Retinopathy Diab Retinopathy No (n = 127) 0.029 (0.032) 0.196 No (n = 192) 6.25 (8.96) 0.014 Yes (n = 66) 0.022 (0.031) Yes (n = 82) 7.98 (11.76) Diab Neuropathy Diab Neuropathy No (n = 159) 0.028 (0.032) 0.613 No (n = 223) 6.25 (10.66) 0.740 Yes (n = 34) 0.023 (0.034) Yes (n = 51) 6.18 (7.79) Peripheral Artery Disease Peripheral Artery Disease No (n = 183) 0.027 (0.032) 0.222 No (n = 262) 6.14 (10.53) 0.287 Yes (N = 10) 0.035 (0.037) Yes (N = 12) 10.78 (12.66) Coronary Artery Disease Coronary Artery Disease No (N = 161) 0.029 (0.034) 0.058 No (N = 234) 6.31 (10.62) 0.871 Yes (N = 32) 0.021 (0.023) Yes (N = 40) 5.49 (8.74) Stroke Stroke No (n = 186) 0.027 (0.032) 0.669 No (n = 262) 6.27 (10.62) 0.369 Yes (n = 7) 0.023 (0.021) Yes (n = 12) 4.92 (9.56)
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Prevalence of type 2 diabetes in Qatari women aged between 18 to 40 from Qatar Biobank
Authors: Abeer Elshewehy and Soha DarghamPrevalence of type 2 diabetes in Qatari women aged between 18 to 40 from Qatar Biobank. 1Abeer Elshewehy, 1Soha Dargham, 2Eric Kilpatrick, 1Lina Ahmed, 1Youssra Dakroury, 1Stephen L Atkin 1Weill Cornell Medicine- Qatar, 2 Sidra Medical and Research Center Background: The Qatar STEPwise surveillance reported that the prevalence of type 2 diabetes amongst Qatar nationals was 16.7% in 2012. This high prevalence of diabetes will translate into significant morbidity and mortality through diabetes complications. Methods: Qatar Biobank extracted data from all women between 18 to 40 years (inclusive), 749 women in total. Women with (Group A: HbA1c ≥ 6.5%; n = 12/708; 1.7%) and without (Group B; HbA1c ≤ 5.6%; 608/708; 85.9%) type 2 diabetes were identified and their demographic details compared. Prediabetes was found in 88/708 women (HbA1c 5.6-6.4%; 11.7%). Biostatistical analysis was undertaken using SPSS. Results: Group A (HbA1c ≥ 6.5%) had worse Framingham Risk Score (2%) than Group B (HbA1c ≤ 5.6 %) ( < 1%). All results are reported as mean value Group A versus Group B. Conclusion: In this small young cohort, 1.7% of women had a WHO diagnosis of type 2 diabetes and 11.7% with prediabtes. Diabetes is perceived to be a disease of the middle aged and older individuals, but this study shows that there is a need for early screening and intervention for the prevention of type 2 diabetes in young Qatari women below the age of 40. Limitation of the study: Small number of participants and that latent type 1 diabetes could not be excluded that may have affected the overall prevalence, though this was expected to have a minimal impact. Acknowledgments: We are grateful to the Qatar Foundation and the Qatar Biobank for their support and assistance, and for Biostatistics, Epidemiology, and Biomathematics Research Core at Weill Cornell Medicine in Qatar
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Tracking health awareness in MENA through Facebook Advertising audience estimates
Diabetes epidemic is claiming more lives than ever, with the number of people with diabetes rising from 108 million in 1980 to 422 million in 2014. World Health Organization estimates that in 2015, diabetes was the direct cause of 1.6 million deaths and in 2012 high blood glucose was the cause of another 2.2 million deaths (http://www.who.int/mediacentre/factsheets/fs312/en/). The obesity and diabetes are especially prevalent in GCC, and Qatar in particular where 70% of the population either overweight or obese (http://www.qatar-tribune.com/news-details/id/54297). The most common form of diabetes – Type II – is caused by a combination of lifestyle factors and genes, thus numerous campaigns are underway to encourage change in daily behaviors which may lead to obesity and diabetes. Recently, Facebook and other social media has become a platform for patient interaction, outreach and education.The scale and reach of Facebook and other social media provide a unique opportunity to track the awareness and interest in health-related topics of large slices of population. This is imperative, as the awareness of an issue is the first step to lifestyle behavior change. In this work, we use the Facebook Advertising platform to track the interests of millions of people across the MENA regions on health-related topics. In particular, before ordering an advertising campaign, the potential advertiser is free to query Facebook about the potential reach, or estimated audience, of a particular selection of location, gender, age, language, interests, and a variety of other attributes. For example, when we select Arabic-speaking women living in Qatar between the ages of 30 and 50 who are interested in dieting, Facebook gives an estimated audience of 4,300 users who would see our hypothetical ad. As such reach estimates do not divulge information on any particular user, this information provides a window into otherwise private behaviors of Facebook users, in aggregate. In the present study, we examine the interest Facebook users from the countries in the MENA region have concerning health-related topics such as Obesity and Diabetes awareness, as well as general Fitness and Wellness and Healthcare. These interests are then compared to baselines such as Luxury Goods and Shopping, as well as health-related ones like Fast Food. We designed a visual analytic interface to enable exploration of these health awareness data. Once the interests are selected (Fig. 1, top), demographic slices of the data are presented in tree maps (Fig. 1, left) and on a choropleth map (Fig. 1, right), with each segment colored with a Health Awareness Score. The user can select a particular demographic (such as male gender) or country, automatically updating all other demographic segments to correspond with the current selection.Fig. 1. Comparison of interest in Physical Activity vs Fast Food for young women who are local to each of the MENA countries. Tool is available online at http://sha.qcri.org/ Using Facebook Advertising, or any other social media, as a source for health awareness monitoring has several drawbacks, the most prominent of which is the biased sampling of the population. However, with the continued adaptation of these platforms across the world and segments of population, they become increasingly representative of the general population. This can be mediated by careful cross-correlation with authoritative data sources such as statistics provided by the World Health Organization. Another drawback comes from the black box nature of the platform, as we are not purview to the internal processing and definitions the platforms use to come up with the metrics they reveal. Our ongoing efforts are addressing this by comparing the metrics provided by Facebook to the known disease prevalence as collected. In particular, if we look at the rates of diabetes (as published by the Institute for Health Metrics and Evaluation (IHME) (http://www.healthdata.org/), we can compare them to the estimates of interest Facebook provides. In Qatar, the Pearson correlation between diabetes rates and Facebook awareness of Diabetes Mellitus Type II across different age groups is r = 0.963 (p < 0.001), suggesting the awareness of disease rises as the incidence of the disease increases in the population. Interestingly, the trend reverses when we correlate interest in Diabetes Mellitus Type I (a much rarer form of disease, usually diagnosed in children) at r = –0.844 (p < 0.002), indicating the focus of the population remains with the Type II. In summary, this demographically rich data source has a great potential for improving awareness campaign tracking and dynamic opinion monitoring. Our study develops new methodologies and tools to collect, verify, visualize, and make this data available to the policy makers, campaign organizers, and many other potential actors interested in monitoring public opinion.
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Vascular endothelial dysfunction as the mechanism for the development of obesity
Authors: Nasrin Mesaeli, Rajaa Saleh Dalloul, Tatiana Lobo and Hamid MassaeliVascular endothelial dysfunction as the mechanism for the development of obesity Raja'a Dalloul, Tatiana Lobo, Hamid Massaeli, Nasrin Mesaeli Weill Cornell Medicine- Qatar, Qatar Foundation, Doha, Qatar Background: Obesity is one of the major public health issues in the world with a rapid increase in its prevalence. According to the last public health report from supreme public health in Qatar in 2012, 71.8 % of the women were overweight compared to 68.3% of men. Among the gulf region, Qatar has the 6th highest rate of obesity in young boys. Moreover, the WHO survey in 2009 showed 70% of the Qatari children were obese because of the nutritional changes and unhealthy lifestyle. Adipocytes are considered the only cells where their size can vary in physiological conditions. Adipose tissues grow as they store excess energy intake. It's well established that adipose tissue is highly vascularized. These vascular networks play a vital role in the adipogenesis process. The vasculature of adipose tissue provide oxygen, growth factors, nutrients, and cytokines to the progenitor cells that are differentiated into pre-adipocytes and vascular endothelial cells. Vascular endothelial cells form the inner barrier of the vessel and is responsible for maintaining the vascular vasodilation and constriction. Defect in the endothelial cell function has been shown to result as a consequence of different diseases such as obesity, diabetes and high blood pressure. An increase or decrease in the generation of the reactive oxygen is one of the main cause of endothelium dysfunction. The fluctuation in the balance of these factors in the endothelial has an influence in the adipocyte cells which is responsible for the formation of fats or fatty tissues which cause an impairment of adipocytes and may lead to obesity. Calreticulin (CRT) is a multifunctional protein that is expressed in the endoplasmic reticulum of all mammalian cells. The main functions of CRT are regulation of intracellular Ca2+ hemostasis and lectin like chaperone. As part of ongoing research in our lab, we have developed a mouse model overexpressing CRT in endothelial cells. One of the phenotypes of this mouse is the development of obesity and type II diabetes overtime. Therefore, the current research was to examine the hypothesis that endothelial specific overexpression of CRT in mice leads to endothelial dysfunction leading to obesity and diabetes. Methodology: A cell targeted transgenic mouse model overexpressing CRT in endothelial cells [will be referred to as (ECCRT+)] was developed in our lab and used in our study. One of the major phenotype of these mice is the development of visceral obesity and diabetes. To characterize these mice phenotype, 4 weeks old wild-type (wt) and transgenic (ECCRT+) litter mate mice were fed with special diet containing either high fat (60%) or low fat (10%) diet for different time points (8-24 weeks). Body weight and blood glucose was measured bi-weekly. At the end time point glucose tolerance test (GTT) was performed to determine the state of diabetes. Epididymal adipose tissues were collected from the wt and ECCRT+ mice. The tissues were embedded in paraffin, sectioned and stained using histological and immunohistochemical techniques to examine the phenotypic changes (shape and size) of adipocyte in ECCRT+ and wildtype mice. Results: Our results illustrated that these mice suffered from endothelial dysfunction. The GTT assay illustrated that ECCRT+ mice developed diabetes at 16 weeks of age when on regular diet (10% fat diet) and high fat diet expedited the development of diabetes. Fat to body weight ratios, and histological analysis of the fat from these wt and ECCRT+ mice showed significant changes in the fat volume, adipocyte size and adipocyte number suggesting a possible association between endothelial dysfunction and adipogenesis which correlate to the obesity and diabetic developed in these mice. Conclusion: Many studies have focused on how obesity induces endothelial dysfunction. Our study is the first to show an important role of endothelial dysfunction in the process of adipogenesis leading to the development of obesity and diabetes. Our data also highlights the importance of an endoplasmic reticulum chaperone in this process. Acknowledge: This project has been funded by NPRP07-208-3-046.
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Retinal blood vessel analysis using IFLEXIS software on fundus images from the Qatar Biobank
Authors: Arnout Standaert, Patrick De Boever, Bart Elen and Rayaz MalikRetinal examination is a diagnostic pillar in ophthalmology for the early detection of eye diseases such as retinopathy (hemorrhage/exudate) and glaucoma (optic disc abnormality). Large population-based studies have shown that retinal vessel morphological quantification may be useful in predicting the development and progression of hypertension, diabetes and cardiovascular diseases (CVD). The rationale for this is that the retinal microcirculatory bed shares similar anatomical and physiological characteristics with the coronary and cerebrovascular circulations. Therefore, quantification of retinal vessel metrics may reveal insights into the development of coronary artery and cerebrovascular disease. Indeed, retinal blood vessel analysis has gained increasing interest in recent years for predicting the development of hypertension, coronary heart disease, stroke and type II diabetes. A number of software algorithms have been used to quantify retinal vessel morphology, but they require manual input and are time- and labor-intensive. Fully automated image analysis allows objective and accurate assessment of retinal vessel parameters. VITO has developed and released IFLEXIS, software for semi-automated retinal vessel analysis, which includes algorithms to determine blood vessel widths, vessel branching and vessel network complexity, integrated in a user-friendly workflow. During the Belgian Economic Mission to Qatar in March 2015, WCM-Q, VITO and Qatar Biobank signed a Memorandum of Understanding to explore the potential of retinal analysis for the early detection of retinal vessel abnormalities in subjects attending the QBB. Fundus images from 774 people attending the Qatar Biobank contained images from healthy controls, subjects with Impaired Glucose Tolerance (IGT), Hypertension (HT) and/or Type 2 diabetes (T2DM). Here, we report the utility of IFLEXIS in a subset of 597 fundus images obtained from 574 persons which could be analyzed. The following parameters were quantified: (i) FD (fractal dimension, using the box counting method), FFD (Fourier fractal dimension) and lacunarity of the vessel network, and (ii) CRAE, CRVE (central artery/ vein equivalent) and AVR (artery-to-vein ratio). Analysis revealed the following mean (range) for this population: 1.39 (1.32 – 1.53) for FD, 2.75 (1.77 – 2.98) for FFD, 1.00 (0.94 – 1.08) for lacunarity, 154.73 (93.79 – 195.67) for CRAE, 233.26 (159.67 – 307.94) for CRVE and 0.67 (0.48 – 0.93) for AVR. Statistical tests for the retinal parameters reveal interesting differences between the studied disease groups. When comparing HT with control subjects, statistically significant differences were found for the CRAE (150.05 ± 2.17 versus 157.88 ± 1.91, p=1.02×10-6) and the CRVE (231.74 ± 3.60 versus 238.28 ± 2.83, p=0.031). Comparing subjects with HT and T2DM with controls also revealed significant differences in CRAE (145.15 ± 3.62 versus 157.88 ± 1.91, p=4.66×10-8) and CRVE (218.47 ± 5.16 versus 238.28 ± 2.83, p=3.31×10-9). This study showed the feasibility of retinal vessel analysis of standard fundus images from the QBB using IFLEXIS. Despite the fact that the fundus images were originally not taken for this purpose, IFLEXIS software can extract novel retinal blood vessel dimensions. As blood vessel analysis has been shown to be relevant for chronic disease prediction, we propose to utilize the baseline assessment to augment the Qatar Biobank database to predict incident disease in follow-up studies. Single retinal features already appear to differentiate subjects with hypertension and/or diabetes compared to controls. We will undertake quantification of a larger image data set and will perform further data analysis to refine both the diagnostic and/or prognostic use of retinal metric analysis in the QBB population. To this end, we will combine retinal vessel metrics with demographics such as age, duration of disease and other metabolic parameters and study the association with whole genome sequence patterns in this population.
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