Qatar Foundation Annual Research Forum Volume 2013 Issue 1

Immune thrombocytopenic purpura (ITP) is an autoimmune disorder, characterized by dysfunctional cellular immunity including the presence of activated platelet specific autoreactive T cells that recognize and respond to autologous platelet antigens. Autoreactive T cells drive the generation of platelet reactive autoantibodies by B cells as well as T-cytotoxic cell-mediated lysis of platelets. Interleukin-18 (IL-18) is a mediator of T helper type 1 cell responses synergistically with IL-12 that initiate and promote host defense and inflammation. IL-18 has a specific binding protein (IL-18BP) which belongs to the immunoglobulin superfamily. In the present study, serum level and messenger RNA( mRNA) expression of IL-18 as well as IL-18BP mRNA expression were measured in peripheral blood mononuclear cells (PBMNCs) of 100 Egyptian pediatric patients with ITP (70 acute and 30 chronic). In addition to this, we recruited 80 healthy volunteers in order to investigate the possible association between the imbalance of IL-18 and IL-18 BP expressions and the pathogenesis of ITP. IL-18 serum level and mRNA expression were not elevated in cases more than in the control group, but IL-18 mRNA was higher in chronic cases when compared to the acute ones (p = 0.031) and there was a good negative correlation between the platelet count and serum IL-18. IL-18 BP m-RNA was slightly elevated in cases more than in the control group (95% Confidence interval = 1.15-2.01). Our results were not supportive for previous findings of elevated IL18/BP mRNA ratio in ITP patients. This could be referred to the fact that autoimmune diseases are complex genetic disorders, therefore further studies on polymorphisms affecting IL-18 gene expression as well as kinetics of IL-18 expression are required to evaluate the role of interleukin 18 and its binding protein in the pathogenesis of ITP.

Fusarium culmorum is a major fungal pathogen of wheat, causing foot and root rot (FRR) and fusarium head blight (FHB). Yield losses are reported as the grain becomes contaminated by mycotoxins. Among the most bioactive compounds are trichothecenes, sesquiterpene epoxides which are able to inhibit eukaryotic protein synthesis and may cause toxicoses on humans or animals consuming contaminated food or feed. Trichothecenes induce apoptosis and may play an important role in the aggressiveness of phytopathogenic Fusarium species towards plant hosts. The aim of this project is to design, prepare and study new natural and natural-like compounds to be applied in the control of F. culmorum mycotoxin production. Particular attention is paid to the selection and preparation of compounds with selective trichothecene B inhibitory activity compared to compounds showing both mycotoxin inhibitory and fungitoxic activities. The first inhibition experiments were performed using compounds belonging to the family of gallic acid, phenylpropanoids and cinnamic derived acids. In vivo and in vitro test and molecular modeling with computational studies were carried out. A straightforward thin layer chromatography (TLC) method and a quantitative LC-MS analysis were used to identify the presence of B trichothecenes and to evaluate the influence of each compound on different F. culmorum culture extracts. Preliminary results indicate that several molecules are able to inhibit the severity of F. culmorum in planta and its growth as well as trichothecene production in vitro. The level of inhibition of 3AcDON range from 67 to 100% under inducing conditions. Fast and effective methodologies for seed dressing were developed using a natural matrix.

Background: The Middle East and North Africa (MENA) region harbors significant proportions of stunting and wasting coupled to surging rates of non-communicable diseases (NCDs). Recent evidence identified nutrition during the first 1,000 days of life as a common denominator not only for optimal growth and development but also for curbing the risk of NCDs later in life. Collaboration between Qatar and Lebanon was initiated to launch the first mother and child cohort study in the MENA region, examining the effect of maternal and young child nutrition and lifestyle characteristics on birth outcomes and growth patterns. The main outcome of this study is to develop evidence-based country-specific nutrition and lifestyle guidelines for pregnant women and young children to ensure optimal nutrition during the first 1,000 days of life. Methods/Design. This is a prospective three-year cohort study. Pregnant women (n=500) in their 1st trimester will be recruited from healthcare centers in Beirut, Lebanon and Doha, Qatar. Participants will be followed up three times during their pregnancy (once every trimester) and six times after delivery (when the child is 4, 6, 9, 12, 18 and 24 months old). In addition, delivery and birth data will be obtained from hospital records. Data collection will include maternal sociodemographic and lifestyle characteristics, dietary intake, anthropometric measurements, and household food security data. In addition, a blood sample will be obtained from the mother during her 1st trimester. Breastfeeding and complementary feeding practices, dietary intake, as well as the growth patterns of children will also be examined. The development of the nutrition and lifestyle guidelines will follow a multistep process using the Delphi technique. Discussion: The developed guidelines will help promote balanced nutrition and health during the first 1,000 days; constituting the foundation of effective interventions not only to ensure optimal growth and development but also to curb the epidemic of NCDs in the region. This study provides a unique opportunity for further follow up in light of the growing field of nutrition and disease risk.

Development of a Web-based Hybrid Registry for Acute Stroke in Qatar Leopold J. Streletz1,2, Saadat I. Kamran1,2, Mohammad Shahzad1, Reggie Cruze1, Naveed Akhtar1,2, Ahmed Elsotouhy2, Ahmed Khattab3, Damian Jenkinson3, 1Weill Cornell Medical College-Qatar, 2Hamad General Hospital and Hamad Medical Corporation, Doha, Qatar, 3Bournemouth University, Dorset, United Kingdom. Stroke is a worldwide medical problem and is one of the leading causes of death and disability for people over the age of 40. In Qatar, although cardiovascular constitutes a major cause of morbidity and mortality, very few stroke studies have been reported. Current standards of care for stroke patients require medical or surgical interventions within 72 hrs of the initial presentation. This is why our research focuses on acute stroke in order to help develop and improve current models for stroke prediction in Qatar. To achieve this in a systematic way, a disease-specific registry for acute stroke needs to be developed. A disease registry is an ongoing, inclusive listing of all individuals with an identified disease from a defined population. It can be used to monitor long-term trends of disease and can also offer clinical researchers an approach to identify particular subsets of patients for research studies. Our registry will also contain patient management parameters which will serve a more practical hospital function of quality assurance in the Hamad Medical Corporation (HMC) hospitals. The development of this hybrid registry is the focus of this presentation. The lead principal investigator of this research has had a prototype Acute Stroke Database on trial in a medical center in Saudi Arabia that showed great versatility and gave easily accessible data for stroke patient management. With minor modification and appropriate security, it can be made available on-line to hospitals and clinics throughout Qatar and will provide an invaluable tool for monitoring clinical data necessary for stroke patient selection. The database will initially be hospital-based and the population included will be all adult patients admitted with acute stroke or a transient ischemic attack (TIA). The stroke database report form which will initially be used as the basis for the registry has been designed with seven specific sub-sections: Admission, Transportation, Stoke Evaluation, Stroke Management, Stroke Classification, Functional Disability, and Registry Summary. Data sources for the patient will rely heavily upon Emergency Department's records and hospital admission records for the patients. It is hoped that the registry will provide extensive clinical information on all acute stroke patients admitted into the HMC system. Using this information, we hope to be able to determine the trends and the relative frequency of specific disease manifestations in Qatar. We also hope that this information will stimulate other hospitals within the region to examine their management routines and thereby serve as a basis for improvements in patient care across the Middle East. ( NPRP No. 6-565-3-141 Award, Cycle 6, 2013 )

Background: MPNs are clonal haemopoietic disorders that are characterized by excessive proliferation of one or more of blood lineages. MPNs include PV, ET and PMF which are associated by the presence of JAK2 V617F mutation in about 90% of PV and 50% of ET and PMF. The molecular workup of JAK2 & related gene mutations were included in WHO 2008 as one major criterion for the diagnosis of Ph- MPNs. Aim: To genetically characterize MPN patients (pts) in Qatar according to the latest(WHO 2008) criteria using molecular studies for JAK2 V617F mutation, JAK2 exons 12-15 & MPL (S505N & W515 L/K) mutations. Methods: Blood samples were collected from suspected MPN cases & DNA was extracted. Allelic discrimination assays were used to evaluate point mutations causing JAK2 V617F & MPL (W515 L/K) mutations. JAK2 Exon 12 was analyzed using High Resolution Melting Curve (HRM) assay & Sanger Sequencing. In some cases the entire MPL exon 10 & exons 12-15 was studied by RNA extraction followed by cDNA synthesis, amplification & sequencing. Results: 400 patients were classified into PV, ET and PM. Out of 180 PV, 97% of cases were positive for the JAK2 V617F mutation and 3% of cases were negative for other mutations. Out of 200 ET, 48% of cases were positive for JAK2 V617F, one had MPL S505N mutation and 50% of cases were negative for other mutations. Out of 20 PMF, 33% of cases were positive for JAK2 V617F and one unclassified case was characterized by DVT had JAK2 exon 13 mutation (R564L). Conclusion: This study used novel molecular approaches to confirm the diagnosis of MPNs cases in Qatar. The observed patterns of mutations were found to be similar to the international data. In our cohorts of patients, JAK2 V617F mutation was found to be present in almost every patient with PV, nearly 50% of ET patients and less than 50% of PMF patients due to the low number of PMF patients in this study. Our original findings show the presence of MPL S505N mutation in one ET patient which was reported both as an inherited or acquired mutation in very rare cases of ET and R564L mutation in one unclassified MPNs case. An ET patient with MPL S505N progressed to AML. The impact of R564L mutation found in 1 atypical case is still unknown & needs further investigation. We report for the first time the presence of most frequent mutations found in MPNs patients in Qatar. This study is preliminary before further molecular investigations to explore & identify mutations in other candidate genes.

Bipolar Affective Disorders are disorders of mood regulation that affect 1-5% of the population worldwide. The severe mood swings that are the hallmark of this disorder can interfere with the patient's personal life and career and often result in marked distress for the patient and family. While the etiology of the disorder remains somewhat elusive, genetic factors are known to be major contributors. Research in patients of European descent have identified several genes that are strongly associated with bipolar disorder. These genes include CACNA1C, ANK3 and Syne1 genes. However these findings have not always been replicated and it is not known whether these or other genes are also associated with bipolar disorder in populations of other racial/ethnic origin. In an effort to address this issue, we recruited 120 patients with bipolar disorder from different Arab countries. The majority were Qataris. All patients were clinically assessed with the Diagnostic Interview for Genetic Studies (DIGS) that we translated into Arabic and adapted to the local culture. The demographic and clinical characteristics of the patient population will be presented. Compared to patients recruited in other parts of the world, our population sample is unique in at least three different and important ways: 1) there is a high frequency of psychosis among our patients (>60%); 2) co-morbidity with alcohol and/or substance abuse is relatively rare (<20%) and 3) about 30% of patients had a family history of a similar disorder in first degree relatives. DNA extracted from a blood sample was obtained on each patient. The DNA was sent for whole genome sequencing at the Hudson Alpha Institute in Huntsville, Alabama in the USA. The sequencing was completed at a depth of 30X. The genetic variants across the genome through all genes, regulatory regions and intronic genetic sequence were encoded in the data. The total amount of data exceeded 10 Tbytes. We have just started the long and tedious process of data analysis that will be shared. These projects lay the foundation for the establishment here in Doha of a repository of clinical and biological data for patients with bipolar disorders. We hope this work will eventually lead to the establishment of the first Arab Psychogenomic Repository for Bipolar Patients in the Middle East.

OBJECTIVE : Taenia solium neurocysticercosis (NCC) is the most common parasitic infection of the brain and is a leading cause of epilepsy in the developing world, especially Latin America,India,Africa,and China .It is increasingly being reported in patients suffering from epilepsy . However, its true prevalence and association with adult-onset seizures is largely unknown in the Middle East particularly in Qatar. Our study will demonstrate that NCC is also a major cause of first seizures in an increasingly young population in Qatar. BACKGROUND Awareness of NCC and associated seizures in the developing countries and the increased frequencies of hospital admissions of young Asian patients with first seizures and abnormal brain CT/MRI suggestive of NCC were the main reasons behind this study. METHODS : This is a retrospective and prospective study, based on hospital populations(2010 to present ) .All patients with seizure(s) seen at the emergency department at Hamad Medical Corporation (HMC) ,the largest single governement hospital in the state of Qatar with a capacity of 1600 beds , were reviewed. Among those patients, all individuals suspected of having NCC were admitted for further investigations and treatment. NCC was diagnosed on the basis of the following : CT or MRI (brain) showing cystic lesions with scolex or Lesions suggestive of NCC on CT/MRI and a compatible epidemiological and clinical history. The complementary examinations included an awake EEG, a CT SCAN or MRI of the brain and serum and CSF studies for some. The results of this study were compared to the most recent data reported in the literature. RESULTS: During this period, 120 patients with seizure(s) were seen at the emergency department .Among them , 55 patients (45,8%) were diagnosed as having NCC on the basis of the above criteria. 54 of the NCC patients (98,3%) were males ;Most of them,53 patients(96.5%), expatriate from the Indian Subcontinent.52 patients ( 94,6%) were older than 20years ; 53 patients(96,5%) presented with seizures with 45 patients (81,8%) having their first seizure. 33 patients ,(60%), had partial seizures, the rest secondary generalized or generalized seizures.17 patients presented with headache. Most of the patients(31) showed a viable cyst on CT/MRI .More than half of patients(29) had an abnormal EEG, however relationship between focal EEG changes and cyst location was only found in 13 patients. Most patients were treated with cysticidal therapy and corticosteroids except those patients with calcification lesions on CT/MRI and absence of edema .Non-neurological side effects included abdominal pain ,nausea, and diarrhea in 8 patients. Antiepileptic therapy (AEDs) was used in 50 patients from the onset and avoided in 5 patients with first seizures and tiny lesions on CT/MRI. During the study-period, AEDs were withdrawn in 6 patients.

Metastasis to the peritoneum and lymph node is associated with poor survival in patients with epithelial ovarian cancer (EOC). To better understand EOC metastasis, we carried out exome sequencing, RNA sequencing and copy number variation (CNV) studies on ovary, peritoneum and lymph node tumors from three patients. To analyze single nucleotide polymorphisms (SNPs) from exome and RNA sequencing data, we use the ratio of reference allele reads to the total number of reads as a measure of a site's allele distribution. This approach is more appropriate for analyzing heterogeneous cancer samples than standard SNP calling methods. Analysis of significant SNP, CNV and gene expression shifts across tumor sites and patients reveals a large degree of heterogeneity between patients. The small number of shifting alleles and differentially expressed genes conserved across patients could be interesting genes for further study. In addition, using both the exome and RNA sequencing data, we identify specific alleles that are expressed at significantly higher and lower levels than expected from the genomic allele ratios. These differentially expressed alleles are very interesting candidates for further study as they may indicate tumor selection for particular alleles. Overall, our results indicate a remarkably heterogeneous landscape of ovarian cancer primary tumors and metastasis and suggest that a more patient-specific research and treatment approach is advisable to achieve better patient outcomes.

Ashraf Al-Amoudi1,2 and Achilleas Frangakis3 1Center of Advanced European Studies and Research (caesar), Department of Molecular Sensory Systems, Ludwig-Erhard-Allee 2, 53175 Bonn, Germany, 2German Center of Neurodegenerative Diseases, Ludwig-Erhard-Allee 2, 53175 Bonn, Germany European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany 3Goethe Universität - Institut für Biophysik Max-von-Laue-Str. 1 60438 Frankfurt, Germany Intercellular adhesion junctions are fundamental for the function and development of multi-cellular organisms. They are widely distributed in animal tissues and most abundant in tissues that are subjected to considerable mechanical stress such as heart, skin and muscle. Desmosomes and adherens junctions (AJs) represent major categories of these junctions. The intercellular space of desmosomes and AJs relies on the associations between members of Ca2+- dependent adhesion molecules called cadherins which share high sequence similarity among them.The intracellular region of the junctions form distinct plaque on the cytoplasmic face of the plasma membrane and form complex network of molecular interactions that link the extracellular region of cadherins with the cytoskeleton. Disruption of cadherins or plaques is the hallmark of many blistering and cancer diseases. Recently, the first crystal structure of the full extracellular domains of C-cadherin (a representative of classical cadherins) showed that the cadherin molecules adopt a stable, curved conformation1. In this crystal structure, the neighbouring molecules are oriented in antiparallel and engaged through a mutual exchange of the tryptophan 2 (Trp2) forming a W-like shape. Such Trp2 trans-interactions were supported by mutagenesis data and cell-adhesion assays and are now considered to be physiologically relevant2. Recently, using cryo-electron tomography and quantitative analysis by sub-tomogram averaging, we revealed the molecular organization of desmosomal cadherins and plaque3,4. Our results show two predominant cis- and trans- interactions alternating in a periodic manner similar to the arrangements observed in the linear zipper of the crystal structure of N-cadherins5. In addition, the resulting molecular model explains previous two dimensional images observed with CEMOVIS at various orientations and yields important insights into the assembly of cadherin-based intercellular junction. Our analysis of the desmosomal plaque revealed two-dimensional interconnected quasiperiodic lattice with similar spatial orgnaization of the extracellular region of the desmosome5. We are currently studying the molecular organization of AJs from mouse intestine. Our results indicate highly-organized structure of E-cadherin compatible with the X-ray structure of classical cadherins1. All these results will be presented in the conference. 1. T.J. Boggon, J. Murray, S. Chappuis-Flam ent, E. Wong, B.M. Gumbiner and L. Shapiro, Science. 296 (2002) 1308. 2. S. Troyanovsky, Eur J Cell Biol 84 (2005), 225. 3. A. Al-Amoudi, D. C. Díez, M. Betts, A. S. Frangakis, Nature 450 (2007), 832. 4. A. Al-Amoudi, D. Castaño-Diez, D. P. Devos, R.B. Russell, G.T. Johnson, A. S. Frangakis,Proc. Natl. Acad. Sci. 108 (2011), 6480. 5. L. Shapiro, A.M. Fannon, P.D. Kwong, A. Thompson, M.S. Lehmann,G. Grubel, J.F. Legrand, J. Als-Nielsen, D.R. Colman and W.A. Hendrickson, Nature. 374 (1995) 327.

Objective: To determine the frequencies of abnormal pregnancy outcomes in a cohort of patients in Qatar and to identify clinical and laboratory factors predicting adverse fetal and maternal outcomes with systemic lupus erythematosus in multinational population in Qatar. Study design: Data of 69 pregnancies of 37 systemic lupus erythematosus (SLE) patients from January 2005 to July 2012 in hmc analyzed retrospectively. Lupus activity was assessed based on SLE Disease Activity Index (SLEDAI) criteria. Results: Among 69 pregnancies 35 (50.7%) pregnancies were in Qatari nationals and 34 (49.3%) pregnancies were in Non-Qatari population including Asians and Africans. There were total of 69 pregnancies from 36 patients. Average numbers of pregnancies prior to and after onset of SLE were 3.73±1.8 and 2.72±1.5 respectively. Age at conception (in years) and gestational age at delivery (in weeks) were 34.5±5.4 and 37.4±2.8 respectively. Anti phospholipid antibodies (aPLs), Anti-SSA (Anti Ro) antibody and Anti SSB (Anti La) antibody were present in 18 (26.1%), 23 (33.3%), 13 (18.8%) pregnancies respectively. There were 10 (14.5%) abortions, 5 (5.7%) stillbirths, 1 neonatal death and 54 (78.3%) live births including two twin gestations. Although, not statistically significant, mean gestational age (weeks) was found to be higher in active lupus patients (37.4±1.9vs. 37.2±3.23, p=0.789) and baby weight at birth was found to be low (2.68±0.64 vs. 2.87±0.60, p=0.360) compared to patients on remission. Pregnancy induced hypertension (PIH) (17.4% vs 11.1%), intrauterine growth retardation (IUGR) (36.4% vs 11.8%), preterm delivery ([31.6 % vs 11.8%), still birth (13% Vs 5.6%) and Eclampsia (13% Vs 0%), all were observed to be higher in active lupus patients compared to patients on remission. However thees differences were not statistically significant. Compared with pregnancies without lupus nephritis (n=44), pregnancies with lupus nephritis (n=7) were associated with a higher risk of still birth (28.6% vs. 4.5%, p=0.092), higher rate of eclampsia (28.6% vs. 4.9%, p=0.103), IUGR (42.9% vs. 26.2%, p=0.626), PIH (28.6% vs. 9.8%, p=0.412). The percentage of live births was higher in pregnancies without lupus nephritis compared to pregnancies with lupus nephritis (42/44, 95.5% vs. 5/7, 71.4%, p=0.092), and live births was also significantly higher in pregnancies without eclampsia/preeclampsia compared to pregnancies with eclampsia/preeclampsia (42/42, 100% vs. 2/7, 28.6%, p<0.001), Stillbirth and preterm delivery were found to be higher in pregnancies with protinurea. Among the laboratory parameters, presence of Anti Ro antibody was found to be significantly associated with IUGR (8/18, 44.4% vs. 6/37, 16.2%, p=0.034). One case of neonatal heart block was found in which Anti Ro/La antibody was positive. Low level of C3 was associated with higher rate of stillbirth, IUGR, preterm delivery, and PIH, however, the difference were not statistically significant (p>0.05). Conclusion: SLE in pregnancy in the Qatar population were associated with higher risk adverse pregnancy outcomes. Disease activity during pregnancy, protinurea, lupus nephritis and eclampsia/preeclampsia were all negatively associated with pregnancy outcome such as IUGR, still births and preterm delivery. Laboratory parameters such as presence of Anti Ro/La antibody and low level of C3 were also associated with adverse pregnancy outcomes.