Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1

Background Dilated Cardiomyopathy (DCM) is a leading cause for heart failure characterized by an enlarged ventricular cavity causing systolic dysfunction. Gene mutations are estimated to be the cause in approximately 30-50% of cases, while modifier genes are thought to influence the clinical outcome. Objectives Using a global and unbiased approach: Next Generation Sequencing (NGS), for one of the largest reported cardiac gene panels, QCRC aims at discovering novel gene variants implicated in DCM pathogenesis and/or progression. Method Through the QCRC Doha-centered intercontinental DCM patient cohort, we screened 38 DCM cases, confirmed by echocardiography, who did not have a history of alcoholism or coronary angiography findings. Patients were recruited following genetic counseling and informed consent, according to institutionally approved ethics protocols. High quality DNA was extracted from peripheral blood, 170 DCM known and candidate genes (=1.6Mbases) were sequenced using the HiSeq Illumina technology, and extensive bioinformatical analysis was pursued to depict genetic variations. Results We herein report an unusual case of a male DCM patient (LVEDD: 70mm; LVESD: 59mm; LVEF: 25%), diagnosed at age 40years, who developed sustained ventricular tachycardia (VT), was implanted an AICD at age 56years, and died age 60years. By NGS we determined that he was heterozygous for a known pathogenic nonsense mutation (c.116T>G) in the phospholamban (PLN) gene, which leads to a premature stop codon (L39X). The phenotype of L39X mutation carriers in the PLN gene has been shown to vary considerably, ranging from severe DCM, to rare reports of hypertrophic CM or normal cardiac function. However, this is the first occasion of L39X DCM patient presenting with sustained VT. Three additional, novel variants, out of the 163 variants detected in this patient, were of particular interest: i) a frameshift mutation (c.1495_1496insAGAC) in the C-terminus of CACNB2 (the beta subunit of the voltage-dependent calcium channel Ca(v)1.2). Mutations in this gene/protein region have been previously associated with Brugada syndrome with shorter than normal QT or early depolarization syndrome. ii) a non-synonymous SNP (c.9217C>T; p.L3073F) in laminin 2 (LAMA2), predicted to have a deleterious (SIFT) - possibly damaging (Polyphen2) effect. LAMA2 is a major component of striated muscle cytoarchitecture, and has been proposed, in a rare occasion, to relate to DCM with ventricular arrhythmias. iii) a non-synonymous SNP (c.6082A>G; p.T2028A) in the Alstrom Syndrome 1 (ALMS1) gene, predicted to have a deleterious (SIFT) effect. ALMS1 is implicated in the development of DCM, with rare reports of cardiac arrhythmias. Conclusion The co-presence of genetic variations in genes such as CACNB2, LAMA2 or ALMS1 may have an important modifier effect to the final clinical outcome towards DCM combined with arrhythmias.

Background and objective: Sleep disorders, such as insomnia can seriously affect an individual's performance and even lead to psychological problems. Diagnosis and treatment of sleep disorders require the collection of subjective and objective measures of sleep structure. Subjective measures are currently collected during face-to-face clinical consultations with a medical practitioner. Objective measures, obtained from polysomnographic (PSG) electrodes on the patient's body, are used to quantify sleep quality, specifically slow wave sleep (SWS) periods. In this work, we present an unobtrusive and ambulatory insomnia monitoring system which uses a single electro-oculograph (EOG) channel instead of a full PSG to identify SWS periods together with a mobile health application to collect subjective measures from patients. Method: Fig. 1 presents the architecture of our proposed system. It consists of three main components. The EOG sensing device records one EOG channel and wireless transmits it to the coordinator. The Coordinator (mobile phone) collects data from the EOG device and transmits it back to the clinical back-end. It also includes a mobile sleep diary application implemented on the Android OS to collect the subjective sleep assessment data from patients. The recorded data is time and date stamped and can be used for historical data analysis. The Clinical back-end consists of a database to store the received data and a server with a SWS classifier to assess sleep quality. Our SWS classifier is based on a simple rule based algorithm using spectral features extracted from several bands (alpha band (8 - 12 Hz), beta band (18 - 30 Hz) and delta band (0.5 - 4 Hz) with adaptive thresholds. We used EOG overnight sleep recordings from nine healthy subjects (18-64 years) to validate our SWS detector. Results: Our developed sleep diary was compared to existing sleep diaries applications. It was found to allow efficient handling of data with an improved layout and interface that enhanced user experience. Information collected through the diary also provided the clinician with better access to the required subjective data for an accurate diagnosis of insomnia. Our developed SWS detection algorithm was run on a testing group with 5 subjects and a validation group of 4 subjects. Sensitivity, specificity, relative observed agreement and Cohen's kappa coefficient values were computed and used to compare the output of the algorithm to the sleep experts' analysis. The agreement of our SWS detection method for the validation data was 90.0%, sensitivity 90.5%, specificity 89.9% and kappa value 0.74. Conclusions: Current PSG systems with multiple electrodes are inconvenient and uncomfortable for the user, resulting in modified sleep activity different from their normal night of sleep. Our work has shown that it is possible to reduce the complexity of the insomnia monitoring experience and still collect the required subjective and objective information needed to assist in insomnia diagnosis. Acknowledgement: The work was supported by NPRP grant #[5-1327-2-568] from the Qatar National Research Fund which is a member of Qatar Foundation. The statements made herein are solely the responsibility of the authors.

Lara Bou Khzam1, Katherine Hajjar2, Nasrin Mesaeli1 1Weill Cornell Medical College in Qatar, Education City, Doha, Qatar and 2Weill Cornell Medical College New York, USA. Diabetic retinopathy (DR) is a leading cause of legal blindness in working-age individuals resulting in disrupted vascular integrity and pathological retinal angiogenesis in both type I and type II diabetes. Annexin A2 is a regulator of endothelial morphogenesis and plays a role in the modulation of fibrin homeostasis and neoangiogenesis by regulating the generation of plasmin. Heterotetramerization of S100A10 (p11) with Annexin A2 and translocation of this complex to the cell surface is required to modulate the activation of plasmin. We hypothesize that the Annexin A2-p11 system regulates the development of retinal angiogenesis and may be a potential therapeutic target in diabetic retinopathy. Objective of current study is to determine the effect of hyperglycemia on Annexin A2 and p11 expression in endothelial cells. To address this objective we examined effect of high glucose on Annexin A2 and p11 expression in HUVEC cells. Our results illustrated no significant change in the total Annexin A2 protein or mRNA after exposure to different doses of glucose for different times. However, hyperglycemia increased p11 protein expression. To determine weather hyperglycemia affects cellular distribution of Annexin A2 and p11 we used cell surface biotinylation and immunocytochemcial staining (ICC) . Biotinylation studies illustrated an increased cell surface expression of both Annexin A2 and p11 under high-dose glucose conditions. ICC experiments also illustrated cell surface localization of Annexin A2 and p11 following high glucose exposure. Overall our data illustrates a role for high glucose in post-translational regulation of Annexin A2 and p11. Future studies are focused on examining the nature of the post-translational modification in Annexin A2 and p11. Acknowledgment: This work is supported by NPRP No: 6-736-3-187 by QNRF.

A consanguineous Saudi Arabian family with two female siblings affected by an autosomal recessive condition resembling Familial Exudative Vitreoretinopathy [FEVR], but also with short stature, bone fragility with thin and wasted appearance was studied by homozygosity mapping and positional candidate gene screening to identify the offending gene and mutation. The gene was mapped to three possible homozygous genomic regions [[2q, 4q, 11q], as the family structure did not allow identification of a single interval with a significant LOD score. Mutations in three genes (FZD4, TSPAN12, NDP and LRP5) have been associated with FEVR. The LRP5 gene localizes within the 11q13.2 homozygosity interval in this family rendering it the positional candidate of choice. Screening by Sanger sequencing identified a novel homozygous one-base splice-site deletion mutation c.3236+1 delG in exon 14. LRP5 is a low-density lipoprotein receptor (LDLR) a transmembrane protein that binds and internalizes ligands in the process of receptor-mediated endocytosis. The cDNA encodes a 1,615-amino acid protein containing conserved modules including a putative signal peptide, four epidermal growth factor (EGF) repeats with associated spacer domains, three LDLR repeats, a single transmembrane-spanning domain, and a cytoplasmic domain. The extracellular domain contains 6 potential N-linked glycosylation sites. LRP5 has a unique organization of EGF and LDLR repeats compared to other LDLR family members and in addition to FEVR, mutations in the gene have been associated with Hyperostosis corticalis OMIM 144750; Osteopetrosis, autosomal dominant 1 OMIM 607634; Osteoporosis-pseudoglioma syndrome OMIM 259770; Osteosclerosis OMIM144750; van Buchem disease, type 2 OMIM 607636; Bone mineral density variability 1 OMIM 601884; Osteoporosis OMIM 166710. Only missense mutations and splice site substitutions in LRP5 have been associated with autosomal dominant and recessive FEVR. This is the first report of an autosomal recessive LRP5 splice-site deletion mutation causing a syndromic form of FEVR.

Toxoplasma gondii (Tg), causative agent of the disease toxoplasmosis in humans is an opportunistic pathogen belonging to the phylum Apicomplexa, which includes many medically important pathogens such as Plasmodium. Tg life cycle is characterized by two cysts stages: tissue cysts containing bradyzoites and oocysts. Taken together tissue cysts in contaminated food/water and oocysts shed into the environment are responsible for nearly 25% of the adult population worldwide being chronically infected with this parasite. Clinically, encystation by Tg presents the greatest challenge towards treatment of Tg as the cyst form is currently untreatable. Our aim is to identify genes and regulatory networks that regulate Tg oocyst/tissue cyst production. Isolation of such global regulators, which regulate encystation will be crucial for designing therapeutic strategies which effectively prevent cyst formation and block transmission of the disease by felines. Using a forward genetics approach, we have isolated a unique Tg mutant, named 11BE9, that under normal growing conditions (tachyzoite stage) in vitro dramatically expresses more than 100 genes, that are usually expressed in the oocyst/bradyzoite stages. Our hypothesis is that a global regulator(s) has been deregulated in this mutant, which is responsible for the over-expression of cyst-associated genes in the tachyzoite stage in the mutant. To identify this regulatory gene, we compared the transcriptome of this mutant to wild type Tg using microarrays, and Tg FIKK kinase gene was identified as a potential candidate. FIKK kinases are novel serine threonine kinases that share a conserved stretch of amino acids and are hypothesized as drug targets due to their unique expression in Apicomplexa. Our preliminary studies show that deletion of the FIKK gene in Tg leads to an impaired in vitro conversion of the tachyzoites to bradyzoites which is required for the establishment of the chronic cyst stage of the parasite during toxoplasmosis. Currently we are in the process of complementing the gene deletion clone to confirm the role of Tg FIKK kinase on cyst formation.

Abstract Gingival tissue-derived mesenchymal stem cells (GMSCs) were recently identified and characterized as having multipotential differentiation and immunomodulatory properties in vitro and in vivo, and they represent new postnatal stem cell types for cytotherapy and regenerative medicine. Dental pulp stem cells (DPSCs) have very low morbidity, high efficiency and extensive differentiation ability. This study aimed at comparing the regenerative potential of pulp derived stem cells and gingival derived stem cells in periodontal alveolar defects through surgically created distal 3-walled periodontal defects with ligature-induced periodontitis were produced bilaterally in the premolar teeth in eight beagle dogs. Simultaneously, DPSCs were derived from the lower precanine teeth of the same dogs, and GMSCs were excised from interdental papilla from each defect site. Three months after creation of the periodontitis model, autologous DPSCs seeded in collagen sponge were implanted on one side as the test group, the other side was implanted with GMSCs seeded in collagen sponge, and unloaded collagen scaffold was used as control group, . Animals were then euthanized and regeneration of the periodontal defects was evaluated clinically in terms of clinical attachment level (CAL), probing depth (PD), and defect size (DS), histologically and histomorphometrically. Variables were compared between groups by pairwise wilcoxom statistical test. Results: All groups show decrease in PD, CAL, and DS with no statistical significant difference between gingival and pulp-derived stem cells groups at the mesial, distal sides as well as the furcation area (P-value = 0.452, 0.785 and 0.539, respectively).These results suggest that both gingival and pulp-derived stem/progenitor cells show significant periodontal regenerative potential.

Dates are important dietary component in the Arab region. Dates production in Qatar is essential for the country's long term food security plan. In this project, we set to investigate the global sources of variation in the dates metabolome. To this end, fully ripened date samples from 138 different varieties were collected from 14 countries including Morocco, Algeria, Tunisia, Libya, Egypt, Sudan, Jordan, Saudi Arabia, UAE, Qatar, Pakistan, US, Iran and Iraq. In total, 402 different metabolites were measured using GC and LC platforms by two different collaborating centers and the resulting data subsequently analyzed using the multivariate analysis software kit Simca. The analysis revealed a global trend whereby certain dates varieties contained more amino acids and less carbohydrates, secondary metabolites, nucleotides and unsaturated fatty acids than other varieties. Integrating this dataset with another set of 30 samples from the same varieties but at the stage of pre and early-ripening revealed that the same metabolic signature underlies the differences between the varying stages of the fruit development process. In summary, at the population level, dates composition differs most according to the progression of the ripening process of the fruit. The local environment and the conditions of harvest and post harvest would directly affect the kinetics of the ripening and while some dates follow full natural ripening process, others might be artificially dried at the start of the ripening process owing to unfavourable weather conditions. Beside environment, genetic factors can also influence the ripening process as it was observed that the dry types of dates possess a metabolic profile similar to that of the soft types at the early phase of ripening. This is consistent with the fact that the dry types of dates typically go through a short ripening phase owing to their low moisture content. The metabolic differences between the soft and dry types of dates may have dietary implication for people with metabolic conditions such as diabetes.

Background: There are many children who present with a problem of Foreign Body (FB) that can be inserted in different orifices of their body like nose, ears or that are ingested in to their gut or being aspirated to their trachea or bronchioles. FB can be even life threating especially in case of chocking and potential of obstructing child's airway, or ingestion of sharp objects, magnetics, or batteries that may contribute to a lot of morbidity. The intervention to retrieve these FB are different according to the content of FB, location, shape, and child's clinical presentation. Therefore we are planning to study the prevalence of FB in children in our community for the purpose to increase public health awareness to decrease these events from happening again. Objectives: The prevalence, characteristics and outcomes of FB in children in the state of Qatar. Methods: It's a retrospective study, to study all children who presented in 2013 with a problem related to FB to Pediatric Emergency Centers (PECS) in Qatar. PECS are the main pediatric emergency center in the state of Qatar with approximately 200,000 visits annually. Results: There were a total of 697 cases of FB presented to PEC last year. 316 FB in alimentary tract, 178 FB in respiratory tract, 85 FB in Ears, 93 FB in nostrils, and 25 FB in conjunctival sac. Majority of patients with FB in respiratory tract required admission to hospital for bronchoscopy. More data will be available on the presentation date Conclusions: Increase awareness of high risk of FB ingestion or aspiration in small children in Qatar.

Ultrasound imaging system is an important imaging modality for the diagnosis of most pathology. However, in certain situations the accuracy of diagnosis can be altered due to the speckle noise that affects these images, which can lead to a misdiagnosis. Ultrasonic speckle is an interference effect caused by the scattering of the ultrasonic beam from microscopic tissues inhomogeneities. To curb this difficulty many despeckling algorithms are being discussed in literature. Several adaptive speckle filters are proposed based on statistics extracted from the local environment of each pixel. These filters smooth speckle adequately, but they do not preserve details efficiently. In this work we aimed to develop an adapted anisotropic diffusion filter based on Perona and Malik method (PM), that can reduce the speckle noise and at the same time preserve the edges. Experimental results were taken from Sheikh Zaid Hospital located in Rabat-Morocco and are considered to illustrate the performance of the proposed technique.

Background: Vitamin D(VitD) deficiency is associated with co-morbidities in the elderly. VitD deficiency remains an under recognized problem in the general population and is poorly defined in elderly patients. In a geriatric population, VitD deficiency has been associated with poor muscular, physical and cognitive physical performance as well as falls and fractures. VitD deficiency is significantly associated with older age and elderly patients who need hospitalization for a longer duration are more susceptible. Advanced age and low exposure to sunlight are the major factors associated with VitD deficiency. VitD also plays a role in insulin secretion and therefore is associated with type 2 DM (T2DM). Earlier studies suggested a significantly higher risk of T2DM in VitD deficient patients. There are no studies in the elderly population in the Gulf region. Therefore, the present study was designed to assess the prevalence of VitD deficiency and the associated risk factors among a geriatric population in Qatar. Objectives: To investigate the prevalence of VitD deficiency among the elderly in Qatar. Design: A retrospective study conducted between April 2010 and April 2012 that involved chart reviews. Settings: All patients in geriatrics facilities including Rumailah hospital, skilled nursing facility and home healthcare services in Qatar. Participants: geriatric patients of age ≥65 years. Measurements: Patient characteristics and outcomes were analyzed and compared according to the severity of VitD deficiency. Correlation of VitD with co-morbidities was analysed. Mean follow-up period was 6 months. Results: A total of 889 patients were enrolled; the majority (66%) was females and the mean age was 74.9±8.7 years. Patient comorbidities included hypertension (76.5%), diabetes mellitus (63%), dyslipidemia, (47.5%), dementia (26%) coronary artery disease (24%) and cerebrovascular accident (24%). The mean baseline serum Vit D level was 24.4±13.5 International Unit; 72% of patients had VitD deficiency: mild (31%); moderate (30%) and severe (11%). Patients with severe VitD deficiency had significantly higher HbA1c levels compared with patients with optimal VitD (P=0.03). High Density Lipoprotein Cholesterol levels (HDL-Cholesterol) were significantly lower in severe VitD deficiency patients compared with optimal VitD patients (p=0.04). There was a positive correlation between HDL-C and Vit D level (r=0.17, P=0.001) whereas, HbA1c levels showed negative correlation with VitD (r=-0.15, P=0.009). Conclusions: A high prevalence of VitD deficiency (72%) was observed among the elderly in Qatar. Lower VitD was associated with higher HbA1c and lower HDL-C levels. Further studies are warranted to evaluate whether VitD supplementation controls DM and low HDL-C levels among the elderly.