Qatar Foundation Annual Research Forum Volume 2013 Issue 1

Background and Introduction—Foodborne illness has been identified as one of the major hindrances to the advancement of health around the world and bacterial pathogens play a major role in this impediment. Although most of the infections are self-limited, different estimates of the cost of illnesses around the world indicate costly episodes with the numbers ranging from $1,600 to $3,000. The global risk of the foodborne pathogens has been exacerbated by globalization of trade and ease of travel around the world. Qatar is where these two factors intersect. Understanding the pathway by which these threats enter the food chain and pose risk to humans will help in developing risk mitigation strategies. In this study we assessed the potential risk of illness from the consumption of mutton contaminated with Escherichia coli O157:H7 in Qatar and identified critical intervention points that would contribute to mitigating its associated risk. Methods—We used the quantitative risk assessment (QRA) methodology using a combination of deterministic and stochastic approaches to address the stated objectives. The QRA approach helps in identifying stages in the production system from farm-to-table that are likely to play roles in mitigating or exacerbating the risk of illness associated with this pathogen (Figure 1). Data on the probability of E. coli O157:H7 in animals, animal products, retail products, and humans were obtained through repeat cross-sectional studies in these populations. Estimates of the adverse health effects were obtained using risk characterization which integrated data on hazard characterization and exposure assessment, including a dose-response model. A Monte Carlo simulation of inputs in the model was performed using the @Risk software (Palisade Software, Newfield, NY, USA) and parameters were obtained using Latin Hypercube sampling. Sensitivity analyses were performed to capture the effect of uncertainty and variability of the different parameters used in the model on the predicted risk of illness. Results—The probability of illness from the consumption of mutton contaminated with E. coli O157:H7 for a healthy female eating at a restaurant range from 7 x 10-3 to 28 x 10-2 depending on the amount of food consumed (Figure 2). However, the risk for the same female eating at home is less (5 x 10-3 to 24 x 10-2). The estimates of illness are three times higher for immune compromised females exposed either at the restaurant or at home. We also evaluated the risk for healthy males exposed at restaurants under similar circumstances and their risk was higher than for females (13 x 10-3 to 44 x 10-2). A similar trend of reduced risk was observed for men exposed at home (9 x 10-3 to 32 x 10-2). The risk of illness due to this pathogen could be significantly reduced for either gender under different scenarios by increasing the roasting of mutton before consumption. Conclusions—This model provides a science-based justification for the awareness about the importance of the probability of adverse health effects due to E. coli O157:H7 in mutton and establishes a scientific foundation for risk managers and public health professionals.

Background: A Canadian study recommends General Practitioners (GP) to use evidence based Clinical Guidelines (CG) when dealing with co-morbid cardiovascular diseases, in particular for the diagnosis and preliminary management of co-morbid Chronic Heart Failure (CHF) and Atrial Fibrillation (AF). Although paper-based Canadian CG exist for the management of CHF and AF, the challenge for physicians is to simultaneously apply multiple independent CG when dealing with patients having cardiovascular co-morbidities. Objective: The objective of this inter-disciplinary research program is to assist physicians in handling co-morbidities through a computerized clinical decision support framework that recommends evidence-based interventions based on the patient's health profile. We target decision support for the diagnosis and treatment of CHF, AF and co-morbid CHF-AF. Approach: We take a healthcare knowledge management approach to develop a Clinical Decision Support System (CDSS) for handling comorbid diseases. Our solution involves the development of institution-specific CP from a combination of CG, and then generate a CP knowledge model using a semantically-rich formalism—i.e. a CG ontology. The CG ontology semantically defines the clinical concepts in order to establish semantic interoperability between multiple CG. Next, we systematically align the ontologically-modeled CG of different diseases to realize a unified knowledge model that derives the evidence based recommendations for handling both single and co-morbid diseases. Our methodology entails the following steps: (i) knowledge identification to derive specialized disease-specific CG from existing evidence-based sources; (b) knowledge modeling to abstract medical and procedural knowledge from the CG; (c) knowledge representation to computerize the CG in terms of a semantically-rich CG ontology; (d) knowledge alignment to systematically synthesize multiple ontologically-modeled CG to develop a unified ontology-based CG knowledge model representing comorbid diseases; (e) knowledge execution to generate patient-specific recommendations, based on patient data, by reasoning over the aligned CG model; and (f) evaluation of the knowledge model and the recommendations produced in response to a range of clinical scenarios. Results: We present the COMET (Co-morbidity Ontological Modeling & ExecuTion) system to provide clinical decision support for three scenarios: (i) Cardiac Heart Failure (CHF); (ii) Atrial Fibrillation (AF); and (iii) co-morbidity of either AF or CHF. COMET is designed for GP in Nova Scotia and is accessible over the web. Evaluation: A pilot study was conducted to assess how well COMET meets the physician's needs to manage co-morbid CHF-AF. Conclusion: In conclusion, this project provides a solution for the complex problem of handling co-morbidities in a CDSS. Our solution is based on semantic modeling of disease-specific knowledge which extends the possibility of scaling up to include additional diseases and aligning their knowledge models to handle even further co-morbid situations. Our CG alignment approach helps (a) avoiding duplication of clinical tasks; (b) re-usability of diagnostic results; (c) determine compatibility of different clinical activities; and (d) standardization of care across multiple institutions. We believe that this project achieves knowledge translation whereby we have successfully computerized and translated paper-based CG so that they can now be operationalized at the point-of-care by GP to handle comorbid diseases.

hospital infection due surface contamination becomes very important field of study in recent years. Such contamination can provide a good environment for different microorganisms (i.e. bacteria, viruses, and fungi) to grow and transmit infectious diseases when they come into contact with human body . Recent studies show that surface contamination is accounted for 100,000 deaths in United States alone. Antibacterial coatings has been utilized in many fields such as healthcare, industrial and homes to prevent microorganism's growth and used in sterilization processes. Typically, Semiconductors photocatalysts are used as antibacterial coatings to prevent hospital infections, these materials has ability to interact with light through oxidative process to release radicals to destroy bacteria and viruses. Conventional Metal oxides Semiconductor such as TiO2, ZnO have drawn much attention during the last few years because of their novel photocatalytic activity, availability, stability, strong oxidative capacity, and low cost , . However, due to their wide band gap and high excitation binding energy they only allow absorption in the UV region of solar spectrum. Nanomaterials offered cost-effective solutions for many environmental problems such as waste water treatment, pollution, and anti-bacterial treatment . . It has been known that pure ZnO exhibits low photocatalytic activity due to rapid recombination of photo-activated electrons and holes. It is expected that doping ZnO with metals will improve the photocatalytic activity and disinfection effect. Here report two step synthesis method of Ag/ZnO Nanoparticle heterogeneous Photocatalyst for anti-microbial treatment in hospitals. Our results indicate that Ag/ZnO exhibits high photocatalytic activity and disinfection effect by extending the absorption to the visible range of solar spectrum and prevent the recombination. The nanostructures of the prepared Ag/ZnO particles have been confirmed using UV-VIS absorbance, XRD, and SEM analysis. Antibacterial properties of the Ag/ZnO nanoparticles were investigated by inhibition testing against E. coli using filter sheet. The results, revealed an obvious zone of inhibition around the Ag/ZnO nanoparticles sheet, suggesting the antibacterial property of the Ag/ZnO nanoparticles. The Ag/ZnO system is tested in the presence of visible light and in the dark. The results indicate that Ag/ZnO nanoparticles exhibit antibacterial activity even in the dark. We attributed the photocatalytic activity of the Ag/ZnO Photocatalyst to the Plasmon surface effect of Ag nanoparticles and interaction of Ag with ZnO where Ag nanoparticles act as an electron sink, promoting interfacial charge transfer and reducing charge recombination. Dastjerdi, R., Montazer, M. 2010. A review on the application of inorganic nano-structured materials in the modification of textiles: Focus on anti-microbial properties. Colloids and Surfaces B: Biointerfaces 79: 5-18. Onaizi, S.A., Leong, S.S.J. 2011. Tethering Antimicrobial Peptides. Biotech. Advances 29:67-74. Hoffmann, M. R.; Martin, S. T.; Choi, W.; Bahnemann, D. W. Chem. Rev. 1995, 95,69

Adoptive cell transfer therapy (ACT) holds a highly promising treatment for metastatic melanoma patients. ACT involves the ex vivo expansion of autologous antitumor reactive tumor infiltrating lymphocytes and their reinfusion into lymphodepleted patients, accompanied by IL-2 administration. ACT demonstrates objective clinical responsiveness in 40-50% patients, including 10-21% complete responses. However, the mechanisms underlying the observed inter-individual variability in response to this type of therapy are not well understood. We applied whole genome-wide association study (GWAS) using Illumina OmniQuad 1M SNP bead array and expanded the SNP coverage to 33M by imputation against International HapMap and 1,000 genomes data sets. The analysis was performed in 208 TIL or PBMC samples undergone adoptive therapy or high does IL-2 therapy at the Surgery Branch, NCI. Comparisons between patients experiencing a complete responder (CR, n=30) versus those suffering progression of disease (PD, n=108) excluding the reminder partial responder patients identified 140 independent association SNPs which covers 14 functional known genes at a threshold p-value < 1 x 10-5. Within the chromosomal crossover range (1000Kb); we observed possible association with 405 annotated genes. The strongest signal was rs11587096, rs4506458 on chromosome 1 and rs7894773, rs7070986 at chromosome 10. Several genes at those loci had known immunological function. Those include TLR5, MIA3, TAF1A, DUSP10, DISP1, HHIPL2, MAF177B and EIF3a, FAM45A, SFXN4, GRK5, RGS10, TIAL1, BAG3, INPPF5 AND SEC23IP on chromosome 1 and 10 respectively. In particular, comparisons between patients experiencing a complete response (CR) versus those suffering progression of disease (NR) identified a polymorphism in the region flanking the exon 2 of the G protein-coupled receptor kinase 5 (GRK5) gene as one the most statistically significant polymorphism. GRK5 can exert immunomodulatory function by promoting the desensitization of chemokine receptors, modulating NFkB signaling and promoting the Wnt signaling, a pathway involved in the development of particular CD8+ T lymphocytes. Possible functional impact of the significant SNP on Chr 10 at transcript level was further validated by analyzing encoded gene exons using Fluidigm technology. In conclusion, we have identified different loci associated with clinic outcome by GWAS study suggesting that genetic predisposition is one of the important component determine immune responsiveness.

Background: Early and rapid onset of obesity in the Qatari population, consequent to expansion primarily of abdominal adipose tissue, is closely associated with inflammation and insulin resistance. Recent interest has focused on microRNA (miRNA), both as biomarkers and mediators, of disease and therefore potentially therapeutic targets. MiRNAs are stable, non-coding species involved in the post-transcriptional regulation of gene expression. Aims: This study investigated the expression of a panel of miRNAs associated with inflammation and insulin resistance in peripheral blood cells of subjects with a range of adiposities and insulin sensitivities. The effect of surgical weight loss on these miRNAs was also investigated in a subset of patients. Method: Non-diabetic Qatari subjects were recruited from patients awaiting weight reduction surgery and from a normal population. Anthropometric measures were recorded. Fasting blood samples were obtained from all subjects, and in a subset after surgical weight loss, for determination of lipids, glucose, insulin and adipokines. miRNA from peripheral blood cells was used to assess their expression using an inflammatory array. Subjects were grouped by body mass index and insulin sensitivity. Results: Three comparisons were carried out: 1. Normal/over-weight (N/OW) versus obese (Ob), 2. Insulin Sensitive (IS) versus Insulin Resistant (IR), and, 3. Before (BWL) and after weight loss (AWL). Despite being matched for age (M 29.5±9.8 vs F 30.1±6.5 years), and BMI (M:36.1±12.9 vs 36.6±10.0 kg/m2) the males had higher systolic and diastolic blood pressure and mean arterial pressure, while levels of both leptin and adiponectin were higher in females. The N/OW, compared to the Ob group, had lower levels of systolic blood pressure, higher HDL-cholesterol, lower leptin, interleukin-6 and higher adiponectin levels. The IS group compared to the IR, matched for BMI, age and plasma glucose, had lower insulin and HOMA index of insulin resistance, as well as leptin and adiponectin. Weight loss was accompanied by a significant reduction in BMI (BWL:41.6±8.6 vs AWL 34.7±8.7 kg/m2), leptin, insulin and HOMA-IR. Three miRNA were significantly different between the different body weight groups. Conversely, 12 miRNA changed significantly when IS were compared to the IR. Weight loss also produced changes in the expression of ten miR species. Conclusion: It is apparent that significant differences in expression of the miR species occur along with changes in metabolic and inflammatory parameters in the different body weight groups, which are further enhanced both synergistically and independently by insulin resistance. Some, but not all, of the metabolic and inflammatory lesions that accompany obesity and insulin resistance appear to be ameliorated by weight loss. The target genes of these miRNAs are currently being investigated to assign possible functionality.

Obesity has increased at an alarming rate over the past three decades. It is likely caused by increased caloric intake combined with genetic predisposing factors and is a major risk factor for type 2 diabetes (T2D). Obesity is linked to chronic inflammation, which was proposed as a cause for insulin resistance and T2D. The obese state creates a microenvironment within the adipose tissue that is susceptible to accumulation of reactive oxygen species (ROS). Accumulated ROS leads to DNA damage and activation of DNA damage response. When DNA is severely damaged and can't be repaired, the cell undergoes senescence, an irreversible cell cycle arrest. Senescence has been linked to aging illnesses and even to obesity with accumulation of senescent pre-adipocytes. A major player in detecting and defending against oxidative stress is PARP-1. PARP-1 activation has been shown to be associated to melanoma senescence, inflammation, obesity and diabetes. The role of PARP-1 in ROS-induced senescence and inflammation of pre-adipocytes has not yet been investigated. Therefore, to study PARP-1 function in pre-adipocytes senescence, we generated and characterized an in vitro experimental model for cellular senescence in primary human and mice pre-adipocytes using hydrogen peroxide (H2O2) as a source of ROS. Our preliminary data shows PARP-1 induction in ROS-treated pre-adipocytes. We are currently using this senescence model to characterize the function of PARP-1 by using gene-silencing approaches.

Obesity is an established risk factor for hypertension. About 1 billion adults are overweight globally and numbers appear to be increasing. Stimulating endogenous activation of brown adipose tissue (BAT) mass that has unprecedented metabolic capacity can combat obesity, reverse insulin resistance and reduce comorbidities. Renalase is a recently discovered flavin adenine dinucleotide (FAD)-dependent oxidase, predominantly secreted into the circulation from the kidney and regulates catecholamine degradation and metabolism. Renalase gene is also present in heart muscle, skeletal muscle and liver cells in humans, and in mouse testicles. Four isoforms of renalase gene has been identified in humans (hRenalase1 to hRenalase4), however only hRenalase1 is detected in human blood suggesting that hRenalase2 to 4 may have alternative function. Unlike the classical amine oxidases which are expressed intracellularly, renalase is present intracellularly and is also secreted into the circulation. Circulating renalase appears to mirror sympathetic tone, a brief increase in catecholamines results in significant up-regulation of renalase synthesis, secretion and activity. Catecholamines are principle activators of BAT thermogenesis and angiogenesis both in rodents and humans. Characterization studies in renalase‐knockout mice shows that these mice have about 25% reduced body weight compared to control, increased circulating catecholamines, are hypertensive, and have cardiovascular defects. Renalase‐knockout mice have activated sympathetic system and possibly have increased BAT activity as a result of elevated catecholamine's in the circulation. Our findings in adipose tissue demonstrate that the renalase gene and protein is expressed in both white and brown adipose tissue depots. Expression of renalase increases in BAT of mice during non-shivering thermogenesis and in diet-induced obese mice. The expression of renalase gene, protein and secretion are significantly higher in differentiated mature brown and white adipocytes compared to preadipocytes. Stimulation of mature brown adipocytes with norepinephrine, the main catecholamine released during cold induced non-shivering thermogenesis in vivo, results in significant increase in renalase expression suggesting the importance of renalase in non-shivering thermogenesis. In white adipocytes, renalase expression is up-regulated in response to treatment with obesity linked adipokines, specifically, leptin and insulin. We report renalase as a novel adipokine expressed and secreted from both BAT and WAT. From our preliminary findings and observations in renalase knockout mice having increased sympathetic activity and circulating catecholamines, we hypothesise "renalase regulates BAT development, thermogenesis and metabolism". Therefore further studies are needed in renalase knockout mice to understand the role of renalase in adipose tissue metabolism which could provide us clues on its role in obesity and insulin resistance. Understanding the mechanisms of renalase action may provide translational data that underpins future treatment in obesity associated cardio-metabolic complications.

Calcium is a ubiquitous signaling molecule involved in various cellular processes, including fertilization, muscle contraction or synaptic transmission. There are two major sources for Ca2+: the extracellular media and intracellular Ca2+ stores, particularly the endoplasmic reticulum (ER). When intracellular stores are depleted, a signal is transmitted from a Ca2+ sensor protein that localizes to the ER membrane (STIM) to the plasma membrane to trigger the opening of the Ca2+ channel Orai. This mechanism, known as store-operated calcium entry (SOCE) activates multiple physiological processes and provides Ca2+ to the SERCA pump to refill the ER stores. To avoid unspecific Ca2+ diffusion in the cytosol, SOCE is a highly localized signal, spatially restricted to dense domains (clusters) created by protein-protein interactions between STIM and Orai. Ca2+-sensitive effectors that respond rapidly to Ca2+ influx through SOCE are therefore in the immediate vicinity of the cluster. We here describe a novel mechanism that expands this Ca2+ signaling microdomain through functional coupling of SOCE, SERCA and the IP3 receptor, to allow efficient activation of Ca2+-activated Chloride channels (CACC) localized away from SOCE clusters. When intracellular Ca2+ stores were emptied using ionomycin, a small CACC activation by SOCE was observed. Conversely, when the stores were depleted following injection of IP3 there was a significantly larger CACC activation (2.5 ± 0.5 nA, n=14 vs 0.08 ± 0.02 nA, n=20). Surprisingly the size of the SOCE current was similar in both conditions, indicating that similar levels of Ca2+ influx lead to different CACC activation. The same mechanism could also be induced when IP3 production was stimulated by lysophosphatidic acid (LPA) and we did show that the cell membrane depolarization induced by LPA required SOCE activation, indicating a physiological function of the process in regulating membrane potential. Ca2+ injection experiments ruled out any increase in the CACC sensitivity to Ca2+, while imaging intracellular Ca2+ levels reflected the amplitude of the Cl- currents Analysis of the localization of the Xenopus CACC (TMEM16a) and of the ER calcium sensor STIM1 indicated that they did not co-localize but rather have a tendency to exclude each other following store depletion. Simultaneous expression of STIM1, Orai1 and SERCA2b tagged with fluorescent markers indicated that the three proteins co-localized in clusters at the plasma membrane following store depletion, creating a very restricted cytoplasmic volume where Ca2+ is increased prior to its pumping into the ER lumen. Finally, GFP-tagged IP3 receptors were not found to be enriched in the clusters. Our results show functional coupling between SOCE, SERCA and IP3 receptors to effectively activate CACCs in response to agonist induced store depletion. Here, Ca2+ entering the cell through SOCE is rapidly taken up into the ER by SERCAs and released through open IP3 receptors close to its target, the CACC. This mechanism extends the function of SOCE to the activation of channels that or not located in the immediate vicinity of the extracellular Ca2+ source without requiring large intracellular calcium events to target distant Ca2+ sensitive elements.

Title: Preliminary Results and Clinical Relevance of Next Generation Sequencing in Hypertrophic Cardiomyopathy Patients in Qatar. Kholoud Al-Shafai 1, Despina Sanoudou 1,2, Paul Barton 3 , Lama Shuayb 1 , Emmeleia Nana 2 , Pothitos Pitychoutis 2 , Rachel Buchan 3 , Roddy Walsh 3 , Stuart Cook 3, Magdi Yacoub 1,3, Mohammed Alhashemi 4 1 Qatar Cardiovascular Research Center (QCRC), Qatar Foundation, Doha, Qatar, 2 Department of Pharmacology, Medical School, University of Athens ,Greece, 3National Heart & Lung Institute, Imperial College London, London, UK. 4Cardiac OPD and Rehabilitation, Heart Hospital, Doha, Qatar. Hypertrophic cardiomyopathy (HCM) is the most common inherited heart muscle disease with a prevalence of 1 in 500 in the general population. HCM exhibits a remarkable clinical and genetic heterogeneity, with over 30 genes being implicated so far. However, the full spectrum of genes and mutations leading to HCM remains to be discovered to help the understanding of genotype-phenotype relationship and its clinical implications. Next generation sequencing (NGS) technologies are powerful tools to study these issues. Here, we utilized NGS technologies to sequence the exomes of over 170 genes including all known HCM genes. Preliminary results of seventeen HCM patients recruited at the Heart Hospital-Qatar is presented, with particular reference to the clinical implications of NGS in early disease detection, diagnosis and disease management. Different previously known HCM disease-causing genetic variants were detected, within sarcomeric genes including TNNI3, MYBPC3, MYL3 and MYH7. Patients carrying those variants were offered genetic counseling and advised for clinical and genetic screening of relatives, leading to the early detection of HCM in a 13 years old boy, coupled with efforts to detect genotype-positive phenotype-negative relatives. Of particular interest are genetic variants found within PRKAG2, NEXN that have not been previously described in HCM and need to be further assessed and screened in relatives. In addition, two genetic variants were detected in SOS1 and RAF1 that were previously related to Noonan Syndrome leading to the diagnosis of Noonan syndrome variants in these patients with cardiac hypertrophy associated phenotype. This is an ongoing program which is expected to yield further valuable information which could influence the management in these patients.

The interest in smart body‐centric wireless networks (BCWNs) for healthcare applications is increasing as clinical acceptance of wireless technology is growing; they potentially combine ease of use with greater independence to the patient. BCWNs are emerging for in‐patient and out‐patient monitoring of ECGs, pulse oximeter, blood pressure, insulin pumps and blood glucose. Coupled with the use of location‐based systems, BCWNs can be applied to ensure the well‐being and safety of aged people in different scenarios with continuous monitoring and tracking. Particularly, Wearable electrocardiogram (W-ECG) recorders are increasingly in use for long-term care for patients with chronic disease, assistive technology for the elderly, risk management for people in rehabilitation, lifestyle monitoring, pre-and post-chirurgical monitoring, and heart anomalies early detection [1]. Devices currently available in the market can typically provide an activity-aware, 2 channels, 3-channels ECG or a 5-channels ECG [2]. However, the systems mentioned are limited in the quality of ECG singles received due to the restrictions on the number of electrodes used. A 12-channels ECG would give a more detailed look at the heart&#39;s three areas (anterior, lateral, inferior), and changes in certain segments of the ECG in the related leads for each area suggest the area of concern. Thus, this work addresses the demand of a wireless activity-aware 12-channels ECG to replace inconvenient wires. Available off-the-shelf programmable sensors is initially applied to test the functionality of the algorithms and also to identify the required components aiding in reducing size and cost. A key point of the successful deployment is a long-term test trial performed within a partnered cardiologist institution, to verify ECGs data against obtained with data from standardized devices. Eliminating wires attached to the patient that usually strap the patient and limit his/her movement and activities would make a patient more comfortable while wearing the ECG system and therefore allow continuous monitoring over a longer period of time. Moreover, the compensation of ECG noise due to the body movement enables the patient to wear the W-ECG while during normal life activities. In this way, a long-term evaluation monitoring the heart activities after a surgery, enabling an early detection of several heart-related diseases or predict heart activity failures can be performed without the patient repeatedly having to visit the doctor or being in bed, with a dramatic improvement of quality of life. Fig. 1 shows measured average power consumption against margins for multi-hop (MH) and single-hop (SH) networks in W-ECG monitoring applications tested on real subject. References: [1] J. Sriram, M. Shin, T. Choudhury, D. Kotz, &quot;Activity-aware ECG-based patient authentication for remote health monitoring&quot;, ICMI-MLMI &#39;09 Proceedings of the 2009 international conference on Multimodal interfaces, Pages 297-304. [2] Shuo Xiao; Dhamdhere, A.; Sivaraman, V.; Burdett, A.; , &quot;Transmission Power Control in Body Area Sensor Networks for Healthcare Monitoring,&quot; Selected Areas in Communications, IEEE Journal on , vol.27, no.1, pp.37-48, January 2009