Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1

Cdc25C is a member of the dual specific protein phosphatase family capable of dephosphorylating both phospho-Tyr and phospho-Ser/Thr residues. In vertebrates, there are three isoforms of the Cdc25 enzyme (Cdc25A, Cdc25B and Cdc25C). The N-terminal (regulatory domain) region of these three enzymes is highly divergent (~20-25% identity) while the C-terminal (catalytic domain) region is more conserved (~60% identity). The Cdc25C isoform is of particular importance because it dephosphorylates Cdc2, which is part of the Cdc2/Cyclin B complex, allowing the cell to transition into mitosis. We want to gain further insight into how Cdc25C interacts with its substrate Cdc2. One way to do this is to utilize the ability of E. coli to heterologously express recombinant Cdc25c. This will allow us to obtain large amounts of the protein for in vitro characterization. First, several E. coli cell lines were tested (each with unique properties; e.g. expression of toxic protein, expression of disulfide rich proteins) for their ability to overexpress Cdc25C. The cell line and expression condition having the highest yield of full-length protein was chosen and purified using a single immobilized metal affinity chromatography (IMAC) column that will bind a 6xHis affinity tag fused to the N-terminus of Cdc25C or a tandem approach that utilizes two affinity tags (an N-terminal 6xHis affinity tag and a C-terminal StrepII tag). The pure protein will then be subjected to a variety of chromatography techniques such as UV-Vis spectroscopy and circular dichrosim (CD) to characterize structural changes the protein undergoes when binding to its zinc cofactor. Site directed mutagenesis will be used to create mutations in the zinc binding region of Cdc25C and these proteins will also be characterized for their ability to bind or not to bind the zinc cofactor.

Abstract Nanobiotechnology has been recently widely applied in multidisciplinary fields including pharmaceutical applications. The last decade has witnessed an increase research interests focused on the biosynthesis of metal nanoparticles from fungi as a natural sources and as bionanofactories. However, silver nanoparticles (AgNPs) are of great importance particularly in medical therapy applications and can be used as antimicrobial agent. The objective of this study was to biosynthesize silver nanoparticles from the soil fungus Curvularia tuberculata and to examine their efficacy against five strains of pathogenic bacteria namely; Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus using agar well diffusion technique. The synergistic efficacy of biosynthesized silver nanoparticles in a combination with commercially used antibiotic Gentamycin against the selected bacteria was also examined. The biosynthesized silver nanoparticles from fungal free-cell filtrate were characterized by using UV-Vis spectrophotometer analysis, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). UV-Vis spectrophotometer analysis revealed a peak at 420 nm indicating the synthesis of silver nanoparticles. FTIR analysis verified the detection of protein capping of silver nanoparticles while SEM micrographs showed that the synthesized silver nanoparticles are dispersed and mostly having spherical shape within the size range between 10-50 nm. The biosynthesized silver nanoparticles possessed a varied growth inhibition activity ranged between 12- 28 mm diam inhibition zones at different concentrations against the tested pathogenic bacterial strains. A remarkable increase of bacterial growth inhibition (25.5-35.5 mm diam) was detected when a combination of silver nanoparticles and Gentamycin was used. A significant increase in fold area of antibacterial efficacy was observed when AgNPs in combination with Gentamycin was applied as compared with the efficacy of Gentamycin alone. The biosynthesized AgNPs did not show any toxicity against human blood. The synthesized silver nanoparticles by the fungus C.tuberculata is promising to be used as an antimicrobial agent in medical therapy due to their broad spectrum efficacy against pathogenic bacteria. Nevertheless, further studies are needed to investigate the activity of silver nanoparticles as antibacterial agent in vivo.

Worldwide 1.24 million deaths occur annually due to road traffic injuries. Additionally, 20-50 million are injured or disabled. Similarly, within the Gulf Cooperation Council countries injury and mortality due to road traffic injury are high. In the State of Qatar, for example, the road traffic fatality rate for the year 2010 is estimated to be 14 per 100,000. Young adult males are disproportionately affected. Road safety risk factor laws namely, seat belt use, driving under the influence of alcohol, helmet use and speed management exist in Qatar but enforcement is lax. An additional new risk factor causing distraction while driving and causing road traffic injury and fatalities is the use of mobile phone whilst driving a vehicle. Currently, in Qatar baseline national data pertaining to seat belt and mobile phone use among drivers is not available. We have thus conducted an observational study to estimate the prevalence of these two variables among vehicle drivers in Doha. Additionally, we will examine and present the associations between phone and seat belt use, and variables such as the gender of the driver, type of vehicle and the time of the day during which the observations were made. Furthermore, policy implications of the study findings will be discussed. Acknowledgement: This work is supported by the Biomedical Research Program at Weill Cornell Medical College in Qatar, a program funded by Qatar Foundation.

One of the key objectives of the surveillance of communicable disease (CDs) is the identification of areas for research and healthcare system improvement. In a recent paper published in Qatar Medical Journal (http://dx.doi.org/10.5339/qmj.2014.9) is shown the delay in diagnosis of CDs of cases reported at The Cuban Hospital (TCH) during 2012 and 2013. Examples of delay in diagnosis was for tuberculosis 61.7 days, acute hepatitis 18.5 days, typhoid fever 17 days, food poisoning 9.5 days, measles 8.0 days and meningitis 3.8 days. These are diseases that require immediate reporting because of their public health importance and some of them are highly transmissible (e.g. tuberculosis and measles). Additional evaluation of patients with tuberculosis admitted at The Cuban Hospital (many reported in Hamad General Hospital and referred to TCH because of bed crisis) from January 2013 to June 2014 (105 patients) show diagnosis delay of 49.5 days (standard deviation 51.9 years) with a maximum figure of 365 days. In patients with positive smear (highly infectious) the delay was 49.7 days (SD 53.5 days) (minimum 2 days, maximum 365 days). The delay was superior in cases with pulmonary tuberculosis (51.3 days (SD 52 days)) than in non pulmonary (36.1 days (SD 51.9 days). Delay in diagnosis of CDs is significant with regard to not only disease prognosis at the individual level but also transmission within the community. The delay could be divided in ¨patient delay and system delay¨. Patient delay refers to the time between onset of symptom and the first contact with a healthcare professional. The system delay refers to the time between this first contact to the diagnosis or confirmation of the disease. The knowledge of the two components of the delay is essential to identify actions to minimize the delay and reduce the probability of disease transmission at the community. In summary data from the surveillance of CDs suggest the need of research to identify the causes of diagnostic delay and subsequently implement actions its reduction, which will contribute to the prevention and control of CDs in Qatar.

Human endogenous retroviruses (HERVs) have been classified into three different classes I, II, III. Betaretrovirus-like sequences, including HERV-K, belong to the class II and contains ten groups, termed HML1-10. It has been reported that an HML-10 sequence, termed HERV-K(C4), is inserted within the gene coding for the human complement C4 in an antisense orientation (Tassabehij et al. 1994). The expression of the HERV-K(C4) RNA has been hypothesized to possibly modulate C4 expression as well as to act as a possible mechanism of defense against retroviral infections (Schneider et al 2001). Recently, it has been demonstrated that low HERV-K(C4) copy number is associated with type 1 diabetes, raising the possibility that this retroviral element contributes to functional protection against this autoimmune disease (Mason et al. 2014). Given the correlation between lower HERV-K(C4) copy number and expression, it has also been proposed that HERV-K(C4) may influence C4 RNA processing and/or have an impact on other endogenous retroviral sequences (Mason et al. 2014). To have more insights into this hypothesis, we identified, lassified and characterized the HML-10 sequences that are present in the human genome GRCh37/hg19 assembly. We used the model-based software Retrotector (Sperberg et al 2007) to search HML-10 sequences in the human genome assembly GRCh37/hg19 and identified 10 HML-10 proviruses in chromosomes 1, 6, 16, 19 and Y. These proviruses were confirmed to belong to the HERV-K (HML-10) group and shown to have the Rec sequence. Out of the 10 identified HML-10s, 5 sequences have both LTRs, 4 have only 3'-LTR and 1 sequence is without LTRs. Age estimation analysis performed on the 5 sequences with both LTRs indicated that they inserted into the human genome >25 Mya ago. Five sequences conserved the primer binding site (PBS) region, all recognizing the Lys tRNA. Translational analysis showed that all proviral sequences have multiple stop codons that preclude their production of functional proteins. Phylogenetic analyses were performed with complete DNA sequences as well as with the Pol amino acid sequences and will be presented. Overall, we have identified and characterized 10 HML-10 sequences in the human genome GRCh37/hg19 assembly. Precise knowledge of all HML-10 sequences will allow further investigation of their physiological and pathological role in autoimmune diseases (Yu & Whitacre 2004), particularly their possible involvement in type 1 diabetes, giving new insights into their understanding. References 1. Tassabehij et al. NAR 22, 5211-17 (1994). 2. Mason et al. Diabetes 63, 1789-95 (2014). 3. Sperber et al. NAR 35, 4964-76 (2007). 4. Yu & Whitacre Trends in immunology 25, 694-99 (2004).

Abstract Introduction Bleeding is a common complication after cardiac surgery. However, lower gastrointestinal bleeding is not usually associated with this type of surgery. Case presentation A 50-year-old man with a history of aortic regurgitation underwent elective mechanical valve replacement under cardiopulmonary bypass. He experienced a complicated intraoperative course involving unexplained cardiac arrest following induction of anesthesia. He also developed two episodes of massive lower gastrointestinal bleeding secondary to mucosal ischemia while convalescing in the cardiothoracic surgery intensive care unit. After unsuccessful attempts to control the bleeding, exhaustion of blood products, and consideration of the high risk of mortality associated with surgery and the possibility of early- and long-term surgical complications, the decision was made to administer two successive doses of recombinant activated factor VII at 60 mcg/kg. Hemostasis was achieved without adverse systemic or valvular effects. Conclusions A favorable outcome was achieved after administration of recombinant activated factor VII, which controlled the patient's severe lower gastrointestinal bleeding. This outcome suggests the need to raise awareness about the use of this drug in dire circumstances when other conventional measures fail or are unsuitable.

Introduction: Intractable bleeding is one of common adverse events after cardiac surgeries. Recombinant activated factor VII (rVIIa) showed efficiency in controlling postoperative intractable bleeding (1). Its usage after cardiac surgeries should be more investigated. Patients and methods: - All patients received rVIIa post cardiac surgeries over a period of two years were included in a retrospective descriptive study. All patients were evaluated for base line laboratory results and comorbidities, operative details, dose of rVIIa received, total chest drains pre and post rVIIa administration, total blood and blood products given pre and post rVIIa, post rVIIa laboratory results, complications of rVIIa, length of stay in intensive care unit. Results: -. We recruited 19 patients with a mean age of 49±18 years who received RVIIa in a dose of 90 mcg/kg. Packed red blood cells transfusion post rVIIa administration was decreased from mean of 10±6 units to 3.7±1.7 units P<0.0001. Chest drains post rVIIa administration were decreased from median of 2000 ml and IQR of 1200 to median of 500 ml and IQR of 500 P<0.1 .No complications related to rVIIa administration were observed. Discussion: - The study highlight the following findings: Efficiency and safety of rVIIa in the control of intractable bleeding post cardiac surgeries. Usage of rVIIa saves blood bank resources, decreases the need of surgical reopening and decreases morbidity. More studies needed to elaborate clinical guidelines for the usage of rVIIa in the management of intractable bleeding post cardiac surgeries. Conclusion: - There is increase need for clinical guidelines for the usage of rVIIa post cardiac surgeries. In our experience at Qatar heart hospital recombinant activated factor VII is safe and efficient treatment to control intractable bleeding post cardiac surgeries. Reference: - (1) Conrad V. Bishop, William E.P. Renwick, Chris Hogan, Michael Haeusler, Annabel Tuckfield and James Tatoulis: Recombinant Activated Factor VII: Treating Postoperative Hemorrhage in Cardiac Surgery. Ann Thorac Surg 2006; 81:875-879.

Background: Small RNAs play an essential role in fundamental biological processes such as differentiation, proliferation, apoptosis and homeostasis. Evidence suggests that small RNAs may also play function in Cancer progression. The conventional in-vitro two-dimensional (2D) monolayer cell culture models are not sufficient to explain the exact mechanism behind tumor invasion and metastasis. The objective of this study is to analyze the role of small RNAs in tumor invasion and metastasis using in vitro 2D and anchorage independent (3D) models in ovarian cancer cell lines. Methods: Total RNA samples were isolated from several 2D and 3D ovarian cancer cell lines. Small RNA libraries were prepared and sequenced using Next-Generation Sequencing. Results: Gene expression levels of 3D culture differ from 2D culture, especially in epithelial and mesenchymal target genes. We were able to identify the predominant pathways that small RNAs regulate in 3D cultures. These pathways were involved in cellular invasion, migration, metastasis, proliferation, and tumor growth. Conclusion: The Results from this experiment suggest that small RNAs play a crucial role in controlling tumor invasion and metastasis in the studied phenotypes. Therefore, small RNAs may offer a critical target for the development of anti-cancer drugs.

Background and Objective: In the Middle East and North Africa, hepatitis C virus (HCV) infection distribution appears to present a wide range of prevalence. The scale and nature of HCV infection exposure in Yemen is poorly known. The objective of this study was to establish the national population-level HCV prevalence in Yemen and to characterize the epidemiology of this infection in the Yemeni population. Methods: We conducted a systematic literature review following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Sources of data included PubMed and Embase databases. Our analysis included all primary studies reporting HCV antibody prevalence or incidence measures in Yemen. Extracted measures were then classified and analyzed on the basis of the study population's risk of acquiring HCV. Meta-analyses were conducted incorporating inverse variance weighting and using a random-effects model to pool summary estimates of HCV prevalence among general population groups. Results: We identified 28 studies providing a total of 46 measures on HCV prevalence and none on incidence in Yemen. Pooled HCV prevalence in the general population was 1.8% (95% confidence interval (CI): 1.05-2.75). Among blood donors the prevalence ranged from 0.2 to 3.0% depending on governorate of origin. The highest prevalence within the general population groups was reported among pregnant women in one study (8.5%) followed by a study of African migrant community living in a Shanty town in Sana'a (5.2%). Among high risk populations, HCV prevalence among hemodialysis patients was 40.0% in 1999 and 62.7% in 2007. Among patients with acute and chronic liver disease it was 74.1%. Conclusions: National-level HCV prevalence in Yemen is slightly higher than that in developed countries, but comparable to several other countries in the Middle East and North Africa. The high prevalence found among high risk groups may indicate limitations in implementation of infection control and blood screening protocols, or possibly dates back to exposures before these measures were expanded. These findings are of particular relevance for resource allocation and HCV public health programming in Yemen. HCV prevention policy in Yemen should focus mainly on prevention and infection control in settings of exposure such as in medical care and among people who inject drugs.

Background It is widely perceived that little is known about the epidemiology of HIV infection among people who inject drugs (PWID) in the Middle East and North Africa (MENA). The primary objective of this study was to assess the status of the HIV epidemic among PWID in MENA by describing HIV prevalence and incidence. Secondary objectives were to describe the risk behavior environment and the HIV epidemic potential among PWID, and to estimate the prevalence of injecting drug use in MENA. Methods This was a systematic review following the PRISMA guidelines and covering 23 MENA countries. PubMed, Embase, regional and international databases, as well as country-level reports were searched up to December 16, 2013. Primary studies reporting 1) the prevalence/incidence of HIV, other sexually transmitted infections, or hepatitis C virus (HCV) among PWIDs; or 2) the prevalence of injecting or sexual risk behaviors, or HIV knowledge among PWID; or 3) the number/proportion of PWID in MENA countries, were eligible for inclusion. The quality, quantity, and geographic coverage of the data were assessed at country level. After multiple level screening, 192 eligible reports were included in the review. There were 197 HIV prevalence measures on a total of 58,241 PWID extracted from reports, and an additional 226 HIV prevalence measures extracted from the databases. Findings We estimated that there are 626,000 PWID in MENA (range: 335,000-1,635,000, prevalence of 0.24 per 100 adults). We found evidence of HIV epidemics among PWID in at least one-third of MENA countries, most of which are emerging concentrated epidemics and with HIV prevalence overall in the range of 10-15%. Some of the epidemics have however already reached considerable levels including some of the highest HIV prevalence among PWID globally (87.1% in Tripoli, Libya). The relatively high prevalence of sharing needles/syringes (18-28% in the last injection), the low levels of condom use (20-54% ever condom use), the high levels of having sex with sex workers and with men who have sex with men (15-30% and 2-10% in the last year, respectively), and of selling sex (5-29% in the last year), indicate a high injecting and sexual risk environment. The prevalence of HCV (31-64%) and of sexually transmitted infections suggest high levels of risk behavior indicative of the potential for more and larger HIV epidemics. Conclusions Our study identified a large volume of HIV-related biological and behavioral data among PWID in the MENA region. The coverage and quality of the data varied between countries. There is robust evidence for HIV epidemics among PWID in multiple countries, most of which have emerged within the last decade and continue to grow. The lack of sufficient evidence in some MENA countries does not preclude the possibility of hidden epidemics among PWID in these settings. With the HIV epidemic among PWID in overall a relatively early phase, there is a window of opportunity for prevention that should not be missed through the provision of comprehensive programs, including scale-up of harm reduction services and expansion of surveillance systems.

Background: Type II Diabetes Mellitus (DM) is one of the leading chronic diseases in Qatar as well as worldwide. However, the risk factors for DM in Qatar and their prevalence are not well understood. We conducted a case-control study with the specific aim of estimating, based on data from outpatients with DM in Qatar (cases) and inpatient/outpatient controls, the association between demographic/lifestyle factors and DM. We also estimated the contribution of these factors to the occurrence of DM cases among Qatari nationals and non-Qatari expatriate population. Methods: A total of 459 patients with DM from Hamad Medical Corporation Hospital (HMC) outpatient adult diabetes clinics and 342 control patients from various outpatient clinics and inpatient departments at HMC were identified using rigorous sampling methodologies and recruited between the years 2006 and 2008. The association between risk factors and DM was evaluated using bivariate and multivariate logistic regression analyses, and for the population at large and for only Qatari nationals. In addition to odds ratios (OR) and 95% confidence intervals (95% CI), we calculated the population attributable risk fractions for demographic and lifestyle factors in relation to DM. Results: Qatari nationality was the strongest risk factor for DM (adjusted OR=5.5; 95% CI=3.5-8.6), followed by higher monthly income (defined as ≥ 3000 riyals, OR=5.1; 95% CI=3.0-8.7), age > 65 years (OR=3.3; 95% CI=0.9-11.4), male gender (OR=2.9; 95% CI=1.8-4.8), obesity (body mass index ≥ 30, OR=2.2; 95% CI=1.5-3.2), no college education (OR=1.7; 95% CI=1.2-2.6), and no daily vigorous/moderate activity (OR=1.5; 95% CI=0.9-2.3). Among Qatari nationals, obesity was revealed as the main risk factor for DM (unadjusted OR=3.0; 95% CI=1.6-5.6) followed by no college education (OR=2.7; 95% CI=1.5-5.1), while consanguinity did not appear to play a major role in predicting DM (OR=1.5; 95% CI=0.8-2.8). Our findings further suggested that eliminating obesity and improving access to education could reduce DM cases by up to one third in the population at large (31.7% and 26.8%, respectively) and up to half (46.9% and 49.3%, respectively) among only Qatari nationals. Promoting physical activity may as well reduce the burden of DM by up to 9.4% in the population at large and up to 17.3% among Qatari nationals. Conclusions: Demographic and lifestyle factors were revealed as the main risk factors for the high DM levels observed in Qatar, with a contribution that appears to outweigh that of genetic risk factors. While further evaluation of these factors among the Qatari population (as opposed to the population at large) is important and of interest, these findings highlight the need to focus short-term DM interventions on addressing demographic and lifestyle risk factors to achieve substantial and timely declines in DM levels.

Background: Negative Pressure Wound Therapy (NPWT) is well-established in the management of infected surgical wounds. However, it is a relatively new modality of treatment for the protection of clean surgical sites. This study was aimed at developing guidelines for the use of negative pressure incision management to prevent surgical wound complications after heart surgery. Methods: In this prospective study, we included 134 patients at high risk for sternotomy wound complications from September 2011 to June 2014. Selection criteria included: obesity, diabetes, smoking and COPD, fragile sternum, bilateral mammary arteries, delayed primary closure and repeated operations. We applied negative pressure of - 125 mm Hg on the wounds of 67 patients immediately following skin closure, the dressing was removed after 5 to 7 days. The results were compared to a control group with matching criteria (n=67) who had standard dressings. The primary end point of the study was the development of wound complications including surgical site infection, hematoma formation and sternal dehiscence within 30 days. Results: 2 of the 67 patients who underwent negative pressure incision management developed superficial surgical site infection within the 30 days postoperative period (2.99%). The wound infection of the two patients settled with regular dressings and oral antibiotics. While in the control group with conventional wound dressings (n=67), 5 patients developed superficial surgical site infection and one patient suffered a deep sternal wound infection (total 6 out of 67 patients, 8.96%). Based on our experience, we have proposed some selection criteria for the use of negative pressure incision management following heart surgery (table1). Conclusions: Negative pressure is potentially beneficial in the reduction of wound complications. There are no published criteria so far for the application of negative pressure on clean surgical sites post cardiac surgery. Therefore, we proposed some guidelines for the use of negative pressure incision management to protect high risk sternotomy wounds.

Background: a 30 year old gentleman was referred to our in thoracic surgery clinic with an incidental finding of an abnormal shadow around the apical region of the heart on the chest radiograph. He was asymptomatic and non smoker and he had no chronic medical illness in the past. He had no specific findings on physical examination. Echocardiogram revealed rounded cyst in the pericardium around the hear apex measuring 5.9 by 4.1cm. Chest CT Angiography revealed no evidence of coronary artery disease. Magnetic Resonance Imaging (MRI) showed a well defined round pericardial cyst close to the left ventricular apex but there was no evidence of infiltration the left ventricle or wall abnormal enhancement. Procedure and Findings: He underwent robotic excision of the cardiac apical cyst. An inflamed 5 by 6cm cystic mass was found densely adherent to the apical pericardium. There were no adhesions or pleural effusion and the lungs and pleural surfaces were looking normal. The cyst was excised and the cavity was irrigated with hypertonic saline. A chest drain was left in the left pleural cavity. Postoperatively the patient had no complications.Bethadine diluted in normal saline was instilled into the pleural space via the chest drain for washing any remains of the infection. The drain was removed after 48hrs and the patient was discharged from the hospital on antiparasitic treatment (Albendazole tablets). Results: the patient had an uneventful recovery from surgery; he was reviewed in the clinic and had no evidence of recurrence of the disease. Histopathology report revealed laminated membranes with few protoscolices consistent with hydatid cyst (Eccinococcal infection). Conclusion : this is an unusual presentation of cystic echinococcosis (hydatid disease) which was excised robotically. Cystic Echinococcosis is a parasitic disease caused by infection with the larval stage of a tiny tapeworms of Echinococcus granulosus which usually grows in dogs (definitive host), sheep, cattle, goats, and pigs (intermediate hosts). Although most infections in humans are asymptomatic, hydatid disease can lead to the development of harmful, slowly enlarging cysts mainly in the liver and lungs. However, it might rarely affect other organs but it often remains unnoticed and neglected for years.

Rational: Heart failure is a common, costly, and frequently fatal disease. One in three cases of heart failure is due to dilated cardiomyopathy. NHE1 expression and activity are increased in this cardiac defect. We have previously shown that elevated activity of NHE1 in the cardiomyocytes induced cardiac hypertrophy in transgenic mice. However, it protected the myocardium following ischemia/reperfusion. This overexpression of active NHE1 elicited modulation of gene expression in cardiomyocytes including an up regulation of myocardial osteopontin (OPN) expression and a decrease of peroxisome proliferator-activated receptor (PPAR)γ. Since transgenic mice phenotype is very often influenced by the integration position of the transgene, we decide to create new transgenic mice to ascertain our previous data and to test the role of modulated genes. Method and results: We have created 5 new mouse lines overexpressing a constitutively active form of NHE1 specifically in cardiomyocytes (KNHE1+). We have evaluated by echocardiography the cardiac phenotypes and function of two of these new mouse lines. Then heart were harvested and submitted to histological, immunohistological and QRT-PCR analysis. Our data showed that in the both KNHE1 studied lines, a number of mice demonstrated heart remodeling identified by a significant decrease in diastolic interventricular septal (IVSd) and diastolic left ventricular posterior wall (LVPW) thickness and an increased diastolic left ventricular internal dimension (LVIDd) (see table 1). Moreover these hearts demonstrated impaired function with a decreased fraction shortening (<21 % vs 31% in wild type mouse) and ejection fraction. However, the proportion of 12 weeks old mice demonstrating this defect is different in both lines 13% (3/24) in line #4 and 58% (11/19) in line#1 vs 0% (0/37). ANP and BNP up regulation in line #1 confirmed heart suffering. Current experiment is designed to perform a longitudinal study examining evolution of these proportions with the age of the animals. Conclusions: We have developed an interesting comparative model of active NHE1 transgenic mouse lines with low and high rate dilated cardiomyopathy identifying evidence of early and late mechanisms. Further studies will be conducted to evaluate the phenotypes of the other mouse lines and the mechanism by which these changes are occuring.

The endoplasmic reticulum (ER) functions as a storehouse for intracellular calcium. STIM1, a calcium sensor, localizes mostly to the ER membrane. Following Ca2+ store depletion, STIM1 forms puncta that localize to the cortical ER and bind Orai1, a plasma membrane Ca2+ channel, to allow Ca2+ influx. This is the predominant pathway for Ca2+ influx in non-excitable cells and is referred to as Store-Operated Calcium Entry (SOCE). Mutations in STIM1 and Orai1 cause severe combined immunodeficiencies and are linked to several forms of cancers. Tight regulation of the levels of STIM1 and Orai1 is crucial for maintaining the subcellular levels of Ca2+ required for its numerous functions. We are interested in mechanisms of post-transcriptional regulation of STIM1. We have developed a system that allows us to test miRNA-mediated regulation by transfecting different cell types using a GFP and a 3'UTR transcriptional fusion and GFP with no 3'UTR as a control. mCherry expressed from the same vector is used as an internal control of transfection efficiency. The stoichiometry of GFP and mCherry levels in the experimental conditions for Stim1 3' UTR was found to be significantly different from those in the control. Knockdown of Ago2, an important mediator of the miRNA pathway, by siRNA restored GFP expression from these constructs suggesting miRNA-mediated regulation. We have identified a miRNA that regulates Stim1 expression in HEK 293 (embryonic kidney), MCF7 (non-metastatic breast cancer) but not in MDA-MB231 (metastatic breast cancer) cell lines. The mechanisms by which this occurs will be discussed.

Background: Breakdown of blood-retinal barrier (BRB) and subsequent hyperpermeability is a cardinal feature of early diabetic retinopathy (DR) and leading cause of blindness. Fatty acid metabolism is implicated in several biological functions via multiple signaling pathways including the synthesis of bioactive metabolites. Our previous studies established 12/15-lipoxygenase (12/15-LOX)-derived 12- and 15- hydroxyeicosatetraenoic acids or HETEs as proangiogenic mediators during DR via disrupting the retinal levels of vascular endothelial growth factor and pigment epithelium derived factor (PEDF). Purpose: 1) To screen the impact of high glucose (HG) treatment on the levels of bioactive lipids including 12/15-LOX metabolites in cultured human retinal endothelial cells (HREC), 2) To test the impact of 12/15-LOX deletion on BRB function and explore the underlying mechanism in particular the role of NADPH oxidase as a major source of oxidative stress during DR. Material and Methods: Liquid chromatography-mass spectrometry (LC/MS) was used in a comprehensive lipidomic screen of HRECs treated with or without HG (30mM D-glucose). Effect of 12- or 15-HETE (0.1 M) on leukostasis and angiogenesis (tube formation) was examined in HRECs transfected with siRNA against the catalytic subunit of NADPH oxidase (NOX2) or the scrambled siRNA or treated with or without the NADPH oxidase inhibitor (apocynin, 30°M). Human leukocytes (PMNs) labeled with lipophilic fluorescent probe were used for leukostasis assay. Migration assay was performed using the Electrical Cell Impedance Sensor (ECIS). Retinal vascular changes were examined by fluorescein angiogram (FA) and Optical Coherence tomogram (OCT) in streptozotocin-induced diabetic wild type (WT) or 12/15-LOX-knockout mice and in mice received intraocular injection with 12-HETE or 12-HETE with PEDF. Western blotting, immunofluorescence and multiplex system were used to test the changes in the levels of inflammatory markers. Results: Out of 126 bioactive lipids screened, 70 were undetected, 47 un-significantly changed, and 9 metabolites were significantly up-regulated by HG compared to osmotic control (5mM D-glucose+25 mM L-glucose). Intriguingly, six metabolites among the 9-increased are the products of 12/15-LOX pathway (15-HETE, 11-HETE, 8, 15-DiHETE, 12-HETE, 17-HDoHE, and 13-HODE). Of these 15-HETE has the highest fold increase (p value= 0.004). Thereafter, we pursued to determine whether the altered metabolites of 12/15 LOX play a role in BRB dysfunction during diabetes. Retinal vascular leakage was significantly reduced in diabetic mice deficient in 12/15-LOX as evidence by FA and less albumin leakage compared with diabetic wild-type littermates. Furthermore, intravitreal injection of 12-HETE per se in normal WT mice resulted in a marked increases in the vascular leakage and expression of inflammatory markers such as ICAM-1, VCAM-1, CD45 as well as the NOX2 compared to vehicle-injected control or to mice received both 12-HETE and PEDF. In vitro, 12/15-HETEs induced HREC leukostasis, migration, and tube formation. These effects were inhibited by the concomitant use of apocynin and by silencing NOX2. Conclusion: Retinal endothelial 12/15-LOX is activated by hyperglycemia causing overproduction of 12- and 15-HETEs. These metabolites are implicated in the pro-angiogenic, -oxidative and -inflammatory effects of hyperglycemia in HREC. Thus, targeting this signaling system represents an attractive therapeutic strategy to prevent and treat DR.

Neuroblastoma is a solid malignant embryonic tumor. It is usually diagnosed in the infancy and rarely found after the age of 10 years. In the current treatment options, CDDP and TOPO are included. Methods: Neuroblastoma (SH SY-5Y) cell death was investigated after treatment with CDDP and TOPO (0.1nM-1μM) using Trypan Blue Cytotoxicity Test and Total Cytotoxicity and Apoptosis Detection Kit (FACS). These data are compared to the drug triggered increase of the intracellular calcium ([Ca2+]i) concentration using live calcium imaging with the calcium sensitive dye Fluo-4 AM. Furthermore, we analyzed changes of gene expression of selected [Ca2+]i signaling related genes such as the calcium binding protein S100A6, IP3 receptors, Ryanodine Receptors, Calmodulin (CALM) and the Calmoduling Binding Transcriptor Activator (CAMTA1). Results: With increasing concentration of either of the two substances the amount of viable cells was reduced after 24h of application. The tests revealed a higher cytotoxicity of TOPO (about 25% cell survival at 0.1μM) compared to CDDP (more than 80% survival). With increasing concentrations the percentage of cells which were in the “late apoptosis” stage increased for both drugs but were more pronounced for TOPO. A similar observation was made when the incubation time for the two drugs was prolonged to 48h or 72h when the amount of viable cells was further reduced. Individual (but not all) neuroblastoma cells increased their [Ca2+]i after the application of either CDDP or TOPO (both 1 μM). Over the period of three to four hours the fluorescence increased about 40%. The increase was time and concentration dependent for CDDP and TOPO, while a concentration of 0.01μM was most efficient to increase [Ca2+]i. CDDP and TOPO influence the gene expression of SHSY-5Y cells, targeting specific calcium signaling related genes. In this context, a concentration and time dependent increase in the expression of S100A6 was observed in the cells treated with CDDP. TOPO showed a more pronounced effect than CDDP in increasing the mRNA expression of S100A6. The expression of ITPR1 and ITPR3 genes encoding the type 1 and 3 of the IP3R were found deregulated in the neuroblastoma cells treated to CDDP and TOPO. The 1μM CDDP exposure for 72h was responsible of ITPR1 and ITPR3 down regulation however; the same concentration applied for 24h transiently increased the mRNA expression of ITPR3. The application of TOPO did diminished concentration and time dependently the mRNA expression of ITPR1 after 24h and 72h exposure and of ITPR3 after 72h exposure, however 24h exposure to TOPO did transiently increased the mRNA expression of ITPR3 for the lower concentrations tested. The expression of Ryanodine Receptor genes RYR1 and RYR3 was found deregulated in the neuroblastoma cells treated to CDDP and TOPO. Conclusion: Time and concentration dependently, the treatment of SHSY-5Y cells with TOPO and CDDP increased the cell death, at clinical relevant concentration. That correlates with increased [Ca2+]i levels and with the deregulation in gene expression of calcium signaling related genes suggesting that [Ca2+]i regulation is involved in the anticancer effects of TOPO and CDDP.

Biocompatible Graphene Oxide with Anchored Zwitterionic Moiety - Synthesis, Characterization and Application Markéta Ilčíková1, Zuzana Kroneková2, Anna Záhoranová2 and Peter Kasák1* 1 Center for Advanced Materials, Qatar University, P.O.Box 2713 Doha, Qatar 2 Polymer Institute, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, Slovakia *Corresponding author: [email protected] The novel biocompatible graphene oxide (GO) bearing zwitterionic moiety was synthesized and characterized. The zwitterionic structures have recently gained attention in biomedical applications as materials for development of ultra-low fouling surfaces. Generally, zwitterions contain both negative and positive charge in their structures; however the overall charge is neutral. That impart them highly hydrophilic performance; the adsorbed water then prevent the cells or bacteria to interact with surface. Graphene is two dimensional filler interesting predominantly due to high electrical conductivity. In oxidized form the electrical conductivity is compromised due to presence of hydroxyl, epoxy and carboxyl functional groups. In this work, the hydroxyl groups were utilized for anchoring the zwitterionic moiety onto the GO surface. The reaction was performed in two steps. First the silanization with (3-mercaptopropyl)trimethoxysilane was preformed to introduce the thiol functionality onto the GO surface that was reacted with sulfobetaine monomer (3-((2-methacrylamidoethyl)dimethylammonio)propane-1-sulfonate) through thiolene click reaction in the following step. The successful modification was confirmed by FTIR and TGA. Compared to neat graphene, the presence of sulfobetaine improved the graphene biotolerability for fibroblasts and pancreatic beta cells. Modification of GO surface with zwitterionic moiety prevents its negative effect on cell viability. Thus the adsorption of cells on the surface and the cell - GO surface interaction is effected. The modified GO will be used for modification of electrode at biochips construction, and the electrorheological response will be discussed as well. Acknowledgement: This publication was made possible by NPRP grant # NPRP-6-381-1- 078 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.

One goal in the field of soft tissue biomechanics is to understand and to model the properties of collagen-fibers in biological tissues such as blood vessels, brain tissues, and skin which undergo growth and remodeling processes. Experimental studies of Bhole et al.\ [1] suggest that the mechanical properties of fibers in biological tissues depend not only on the current state of the deformation but also on the deformation history. Demirkoparan et al.\ [2] developed a hyperelasticity based framework taking this effect into account. The framework developed in [2] uses an additive hyperelastic material model for the strain energy density, $W = W_m + W_f$. The matrix contribution $W_m$ does not take time-dependent changes of the material properties into account yet the fiber contribution $W_f$ changes over time as it is modeled in terms of a constant creation rate and a deformation dependent dissolution rate of the fibers. Then Topol et al.\ [3] have investigated the predictions of such a model by studying the effect of fiber deformation state at the time of their creation on the overall mechanical properties of the material. This research investigates the relation between the change in the fiber density, the Cauchy stress development, and the deformation of the material.

Diabetic retinopathy (DR) is a leading cause of blindness among working diabetic patients worldwide. The current therapeutic strategies are limited by several side effects. Thus, it is important to investigate novel approaches for therapeutic intervention. Early inflammatory response and late neovascularization (NV) are cardinal clinical signs of the DR. We have previously demonstrated the involvement of bioactive lipids especially the products of omega-6 polyunsaturated fatty acids (PUFAs) such as arachidonic acids in the development of retinal hyperpermeability and NV during DR. Hyperglycemia is a major risk factor for endothelial cell activation and development of microvascular dysfunction during DR. Overwhelming evidences suggest that omega-3 PUFAs like DHA (C22:6n-3) are protective in proliferative DR. In the present study, we investigate the effect of exogenous supplementation of DHA on human retinal microvascular endothelial cells (HREC) under hyperglycemic conditions. Cells were treated with or without high glucose (HG, 25 mM) for 36 hrs in the presence or absence of the DHA (30 µM). To study the early inflammatory response, cells were analyzed for cell barrier function by measuring the changes in transcellular electrical resistance (TER) using the Electrical Cell-substrate Impedance Sensing (ECIS). Hyperglycemia resulted in a significant decline in TER while it was significantly improved by DHA. Furthermore, to investigate the effect of DHA on the proangiogenc properties of hyperglycemia in HREC a scratch-wound repair assay was carried out in cultured HREC cells treated with or without HG in the presence or absence of DHA. The migration and proliferation of cells induced by HG was reduced on supplementation of cells with DHA. Levels of apoptosis and oxidative stress were also monitored under the effect of hyperglycemia. While HG treatment did not result in any change in level of apoptosis, it resulted in sharp induction of oxidative stress in cells, which however was ameliorated under the effect of DHA treatment to hyperglycemic cells. Real-time PCR revealed significantly enhanced transcripts for the vascular endothelial growth factor (vegf) in hyperglycemic cells while reduced expression of the pigment epithelium derived factor (pedf) and the peroxisome proliferator activated receptor gamma (PPAR) was observed. These changes were prevented by DHA. Anti-inflammatory properties of DHA were further evinced by monitoring the levels of several inflammatory cytokines and angiogenic factors by multiplex enzyme-linked immunosorbent assay. Glucose treatment resulted in upregulation of various pro-angiogenic factors like angiogenin, angiopoietin, pdgf, pigf and this effect of HG was prevented by DHA treatment. Similarly, inflammatory cytokines like TNF-alpha and MCP1, which are involved in promotion of angiogenesis and inflammation were induced by HG and was attenuated in the presence of DHA. Interestingly, anti-inflammatory cytokines like IL-1 beta and IL-6 were markedly increased by DHA as compared to HG alone. These results suggest that DHA protects HREC against the pro-angiogenic, -inflammatory and -oxidative effects of HG and thus it can be proposed as alternative or supplementary treatment to the current therapeutic strategies of DR.

General consensus exists that biomedical research is a key determinant of innovation, health, and economic wellbeing. However, the answer to the question how do we determine and quantify the value and the impact of research is apparently simple, while a thoughtful and analytical answer quickly becomes very probing and difficult to address. Qatar has dedicated itself to transitioning from a resource-based to a knowledge-based economy and Sidra Medical and Research Center is envisioned to become a beacon of learning and a world-class institution of its kind in the MENA region. A unique opportunity exists at Sidra for the development of a comprehensive system to capture the return on investment (ROI) for research on knowledge, the economy, and on health outcomes. This would enable Sidra to establish itself as a leader in the field of impact assessment, ahead of many other countries also grappling with this thorny problem. In fact, Sidra's status as a startup creates a rare situation where measurement and analysis of ROI can be initiated from a baseline. Work can then be done prospectively rather than trying to gather information retroactively, a difficult obstacle that institutions that have attempted to implement similar systems have had to try to overcome. In order to take advantage of this opportunity at Sidra, we propose creating a core system consisting of a smart, web-based, auto-populated "Sidra Biosketch". This system will interact with internal enterprise resource planning, grants and proposal management systems, as well as external publication, patent, startup and commercial license databases (e.g., PubMed). This system takes into consideration two critical factors. First, it minimizes the administrative burden on investigators and administrators. Second, it does not solely measure stochastically by counting numbers of publications, citations, etc. Rather, it focuses and relies heavily on content and contextual assessment to enable Sidra to fully understand exactly what research is being done, where it is being done, by whom, and with what collaborators. The impact on knowledge, economic and health outcomes can then be mapped through dynamic network analyses, natural language processing, and topic modeling approaches. This approach to measuring the impact of Sidra research will maximize efficiency of input and analysis as well as the quality of outputs and results. Meanwhile, this system will also enable Sidra to make decisions that will enhance resource allocation and allow for better assessment of its contribution to progress in science, the economy, and health outcomes in Qatar and worldwide.

Nonlinear continuum mechanics is commonly used to study the response of highly de- formable materials. Nowadays, the proposed material models often include internal ma- terial variables in order to capture realistic engineering behavior. The theoretical back- ground is mature and its implementation in the frame of nite element method is well established. Solely, these material constitutive models are expressed in terms of defor- mation gradient F, its derived quantities, and various multiplicative decompositions such as F = ^F F (1) This multiplicative decomposition serves to pertain particular portions of F to specic parts of the material response. The usual scheme is then that ^F models elastic response and it is associated to the rules of variational calculus. The F portion then models inelastic response, usually by means of a time dependent evolution law. Recently, the arguments of variational calculus have been applied to both portions of the deformation gradient decomposition for incompressible hyperelastic material [1]. The decomposition itself is then determined by an additional internal balance equation that is generated by such a variational treatment. The internally balanced compressible hyperelastic material response is presented in [2]. Fundamental studies of this type are key to better physics-based modeling of complex biomechanical processes in soft tissue, including long time scale processes of growth, and short time scale processes of wound healing. The FE formulation of this internal balance multiplicative decomposition is naturally achieved by linearizing the weak form that is obtained by the variation with respect to both portions of the multiplicative decomposition. In this work, we would like to present the Updated Lagrange Formulation. This formulation will be implemented into com- mercial nite element package FEAP that will facilitate in future studying engineering application in the eld of biomechanics. The correctness of implementation will be ex- amined by simulating homogeneous deformations such as uniaxial loading, dilatation and simple shear. References [1] H. Demirkoparan, T. J. Pence, and H. Tsai. Hyperelastic internal balance by multi- plicative decomposition of the deformation gradient. Archive for Rational Mechanics and Analysis, 2014. DOI: 10.1007/s00205-014-0770-9. [2] A. Hadoush, H. Demirkoparan, and T.J. Pence. Modeling of soft materials via mul- tiplicative decomposition of deformation gradient. In USNCTAM, 2014.

Background Islet cell death is a common feature of type 1 diabetes (T1D), including after islet cell transplantation, as well as of type 2 diabetes (T2D). As of today, we lack tools to quantitatively detect islet cell loss prior to the clinical onset of diabetes, or to predict the progression of established diabetes. Our preliminary data demonstrates that a signature of 20 different non-coding (nc) RNAs (including microRNAs) reflects beta cell death (RAPID: RNA-based Analysis for Prediction of Islet Death signature). Objectives 1)To identify a high-throughput platform for detection of ncRNAs/microRNAs 2)To validate the RAPID signature of 20 ncRNAs using this high-throughput platform 3)To test the suitability of these ncRNA biomarkers in predicting the progression to T1D Method Next Generation Sequencing (NGS) studies on developing human pancreas have led to the identification of specific ncRNAs that are found to be expressed specifically in the human pancreatic islets. We use combined FISH and immunostaining to confirm the localization of each miRNA in human islets. TaqMan-based real-time PCR on plasma from at-risk diabetic individuals and age/gender matched controls in a longitudinal study, is used to measure the abundance of specific ncRNAs in plasma-EDTA samples of individuals at risk of T1D. I will discuss the comparison of 2 different ultra-high-throughput TaqMan real-time PCR platforms that we use to analyze these samples and present the data demonstrating the suitability of these RNA-based biomarkers in predicting the progression to T1D. Results Present study has allowed us to validate a microRNA-based signature of islet cell death in diabetes. Using a cohort of kids at risk of T1D, we demonstrate the suitability and advantages of these ncRNA biomarkers over traditional / conventional antibody- and glucose-based detection. The development of an assay for early detection of islet injury and death has direct applications in clinical medicine. Hopefully, this study results will inform medical researchers as to how to predict the development of Type 1 diabetes, monitor response to interventions such as islet transplantation, vaccines and drugs that aim to retard beta cell loss or promote beta cell regeneration.

Over the last two decades the prevalence of obesity has reached epidemic levels worldwide. Secondary major health risks, such as hypertension, insulin resistance, type 2 diabetes and cardiovascular diseases, can be summarized as the metabolic syndrome and are highly associated with obesity. In cases in which energy intake e.g. through high caloric food intake exceeds energy expenditure the overall energy homeostasis is imbalanced. White adipose tissue depots mainly act as the body's main energy storage and additionally act as an endocrine organ by releasing adipokines and cytokines into the body. Additionally, functional brown adipose tissue (BAT) has been discovered through radiological detection in the last years in substantial amount in adults. Formerly overlooked BAT, which was thought to be absent in the human adult, has therefore recently become an interesting anti‐obesity target due to its ability to dissipate energy in form of heat. We extensively characterized human brown PAZ6 adipocytes in comparison with a white adipose cell line SW872. In addition, human SGBS adipocytes were included in the analysis. Brown and white adipocyte markers were tested by quantitative Real-time PCR. Fluorescent and Oil‐Red staining assessed the quantity and quality of the differentiation process. Next generation RNA sequencing of undifferentiated and mature SGBS and PAZ6 cells was performed in order to elucidate pathways distinctly activated in white vs. brown human adipocytes. Functional assessment of oxidative rates of each cell line was conducted using the Seahorse technology. Whereas PAZ6 and SW872 cells showed classical molecular and phenotypic markers of brown and white adipocytes, respectively, SGBS cells presented a versatile phenotype of adipocyte. 14 days after initiating the differentiation process the expression of classical brown marker such as UCP‐1 and PPARγ peaked and declined until day 28. The white adipocyte marker Tcf21 however, showed reciprocal behavior. Interestingly, Leptin levels peaked at day 28 whereas the highest adiponectin mRNA levels were monitored at day 14. Phenotypic analysis of the abundance and shape of lipid droplets were consistent with the molecular patterns. On day 14, SGBS cells showed multiple small droplets, however the number of droplets decreased and the size increased until day 28 as expected for a white adipocyte phenotype. Lastly, functional metabolic analysis showed the highest oxidative rate of mature SGBS cells on Day 14, which remained consistent or slightly decreased until day 28. SGBS cells are widely used as a model for white adipocytes. Our data suggest that the cell line harbors a versatile phenotype, which changes throughout their mature stage. Many reports suggest recently that the traditional classification of adipocytes is not exclusive and the existence of beige/brite adipocytes has been shown. We are presenting data derived from a human cell line model, which harbors characteristics of both distinct phenotypes. This knowledge will be of importance for studies aimed to increase brown fat depots in order to increase energy expenditure in obese subjects with the ultimate goal of weight reduction.

Background: Polycythemia Vera (PV), Essential Thrombocytosis (ET) & Primary Myelofibrosis (PMF) are Philadelphia negative Myeloproliferative Neoplasms (MPNs) characterized by overproduction of one or more myeloid cell lineages. MPNs are associated with the presence JAK2 V617F mutation in 95% of PV & 50% of ET & PMF patients. Several molecular methods such as RQ-PCR, HRM & Sequencing are currently used to detect common mutations. However, there are still significant numbers of MPNs are negative to the most common genetic anomalies. The advent of Next Generation Sequencing (NGS) gives the opportunity to study relevant mutations in several genes. Aim: Utilizing NGS to identify potential genetic anomalies causing familial MPNs patients in Qatar. Methods: 6 MPNs patients from consanguineous families & 5 healthy individuals were consented into the study gDNA was extracted & used for multiplex amplification of amplicons targeting cancer associated mutations in 28 key genes using the Ion AmpliSeq Kit. NGS was performed via the Ion Torrent using the 318 chip & data was analyzed with the Torrent Suite. The confirmation of NGS data was done by RQ-PCR or Sequencing. Results: NGS identified novel deleterious mutations in MPNs patients. Out of 6 familial cases, 5 patients (P1- P5) were ET & 1 patient (P6) was PV. P1 had JAK2 V617F, ASXL1 T600P, CBFB G180S, THPO S184R & ITGA2 R76Q, P2 had JAK2 V617F, MPL A554G & ATM F582L, the other three Patients (P3, P4 & P5) had CLAR K385fs*47 & one PV patient (P6) had TYK2 E1163G, ASXL1 P808H, PDGFRB P4L TERT G300fs. In patients & healthy individuals, mutations/SNVs such as MPL P106L, K553N, SH2B3 L476F, ATM F1036F KIT N564S & TET2 T730R were also found Discussion & conclusion: In this study, several deleterious somatic/germ-line mutations/SNVs were identified using Targeted Exome Sequencing approach. A complex combination of mutations occurred in ET patients & coexistence of several oncogenic events occurred in PV patient. This finding may also suggest that the MPNs phenotype may depend on presence of other mutations. It is worth to mention that the presence of ATM variant in P2 is associated with increased risk of CLL. Somatic CALR type-2 mutation was identified in 3 ET (nonmutated JAK2 or MPL) patients. This mutation is 5-bp TTGTC insertion in exon 9 that generates a mutant protein with a novel C-terminal (p.K385fs*47). In patients & healthy individuals, a heterozygous germ-line mutation in exon 3 of the MPL gene (MPL P106L) has been observed and previously described as a rare autosomal-dominant disorder. However, this mutation is considered to be frequent in Arabic populations also several unreported variants of uncertain significance were identified. Our finding suggested that familial MPNs patients in Qatari tribes have several potential disease- associated variants/mutations which represent a unique interest for the scientific community worldwide and present an unprecedented opportunity for local and international collaborations to carry out WES in society with its unique coherence and genetic pool (founder effect), this would offer invaluable resources to unravel the genetic etiology and pathogenesis, delineate the mechanisms behind MPNs phenotypic diversity.