Qatar Foundation Annual Research Forum Volume 2012 Issue 1

The cardiac muscle contraction is regulated by a set of proteins known as sarcomeric proteins, which are components of thick and thin filaments. Mutation in these proteins especially cardiac myosin binding protein-C, a multi-domain (C0-C10) protein, is one of the major causes of hypertrophic cardiomyopathy (HCM). However, structure-function relationship of this protein is unclear. Mutation E258K, which is located in the C-terminal of domain C1, has been shown to be associated with HCM in Egypt. Hence, the purpose of the study was to understand the molecular basis of this missense mutation. Molecular modelling study was performed using the available crystal structure of the domain C1. Here, we have carried out molecular dynamic simulations for both WT and E258K for 10 ns, and the structures that showed major changes were considered for further analyses. Our results suggest that the mutation can change the local structural stability through altering a series of intramolecular interactions. Moreover, as the mutation results in replacement of a negative amino acid by a positive one, thus, it affects the surface electrostatic properties of the domain (Figure). Hence, it might interfere with the binding to neighbouring domains and with the other sarcomeric proteins such as actin and myosin. E258K appears to affect the structural integrity of the domain C1 both directly and indirectly. It is hoped that these findings can help to understand the molecular mechanism of the disease.

Background and Objectives: Diabetic nephropathy (DN), a serious complication of diabetes, is characterized by hyperfiltration, hypertrophy, extracellular matrix accumulation, fibrosis and proteinuria leading to loss of renal function. In renal hypertrophy, tubules increase in size and cause accumulation of the extracellular matrix, and are also associated with alterations in renal sodium handling as well as hypertension; processes linked by involvement of the arachidonic acid (AA) metabolites 20-HETE and EETs. This study aims to determine the specific AA-metabolizing CYP450 isoforms present in proximal tubules (PT) that are altered by high glucose (HG) in cultured PTs, and in an animal model of diabetes. It intends to investigate the effects of alterations in CYP isoforms and/or AA-metabolite levels in DN. This work will investigate the mechanism of PT injury and the effect of inhibition of AA-metabolites in vitro and will also provide insight into the cross-talk between CYP450 isoforms and other sources of reactive oxygen species (ROS). Methods: Immunohistochemistry, hypertrophy, apoptosis, fibrosis, ROS generation, 20-HETE and EET formation, CYP4A and Nox protein expression, and mRNA levels were measured in vitro and in vivo. Results: Exposure of PT cells to HG resulted in apoptosis and hypertrophy. HG treatment increased ROS production and was associated with CYP4A and CYP2C upregulation, 20-HETE and EETs formation, and Nox oxidases upregulation. The effects of HG on Nox proteins and mRNA expression, matrix protein accumulation and apoptosis were blocked by HET0016, an inhibitor of CYP4A, and were mimicked by 20-HETE. Inhibition of EETs in vitro promoted the effects of HG on cultured proximal tubular cells. In parallel, the levels of CYPs 2B, 2C, and 4A were assessed in a rat model of streptozotocin-induced diabetes. There was significant induction of expression and activity over control of these CYPs associated with an increase in ROS production, Noxs expression, PTs injury, and this was prevented by insulin therapy. Conclusion: Our results indicate that hyperglycemia in diabetes has a significant effect on the expression of AA-metabolizing CYPs, manifested by increased AA metabolism, and might thus alter kidney function through alteration of type and amount of AA metabolites; this pathway is through an oxidative stress-dependant mechanism.

Background: Although the linkage between germline mutations of BRCA1 and hereditary breast/ovarian cancers is well established, recent evidence suggests that altered expression of wild type BRCA1 might contribute to the sporadic forms of breast cancer. The mechanism underlying the downregulation BRCA1 expression is not well understood. It might be dependent upon repressor activity that governs histone acetylation and DNA accessibility at the BRCA1 promoter. TNRC9 (TOX3) gene, highly associated with breast cancer susceptibility, encodes a nuclear protein of uncertain function but can modify chromatin structure. We hypothesized that constitutive expression of TNRC9 could be relevant to breast cancer biology through the modulation of BRCA1 activity. Methods: We assessed the associations of TNRC9 gene amplification with breast cancer onset and survival. The search for targets and effects of TNRC9 was performed using multifaceted molecular approaches. Results: The TNRC9 gene is often amplified and overexpressed in breast cancer, particularly in advanced breast cancer. TNRC9 gene amplification is associated with reduced disease-free and metastasis-free survival rates. TNRC9 significantly increased breast cancer cell proliferation, migration and survival of exposure to apoptotic stimuli, and tumor progression both in vitro and in mice models. Gene expression profiling, protein analysis and in silico assays of large datasets of breast and ovarian cancer samples suggested that TNRC9 and BRCA1 expression are inversely correlated. TNRC9 binds not only to BRCA1 promoter but also to the CREB complex, a BRCA1-transcriptional regulator. TNRC9 downregulates the expression of BRCA1 by altering the methylation status of its promoter. Conclusions: Our study unveils a molecular basis for a TNRC9 role in breast cancer and highlights a new paradigm in BRCA1 regulation.

Background: Ovarian cancer is the second leading cause of cancer-related death in women worldwide. Despite optimal cytoreduction and adequate adjuvant therapy, many patients will experience disease recurrence. Targeted therapies have been evaluated in ovarian cancer as a method to overcome resistant disease. Antiangiogenic therapy such as bevacizumab (Avastin; Genentech) has limited efficacy. Indeed, it increases hypoxia, drug resistance and, tumor rebound phenomenon observed after withdrawal. Hypothesis: We hypothesized that abnormalities in the tumor endothelium contribute to tumor growth which may be a direct source for chemotactic factors and might be responsible for residual microscopic disease and rebound effect following antiangiogenic treatment. Methods: Using a feeder-free Matrigel and spheroid models of ovarian cancer, we examined the effect of bevacizumab on residual disease. We used Akt-activated endothelial cells (EC) that replicates tumor endothelial biology and controls tumor growth, and human umbilical vein endothelial cells (HUVEC) to investigate the antiangiogenic activity of bevacizumab by angiogenesis and migration assays. We conducted an XTT assay to examine the effect of bevacizumab on proliferation of vascular endothelial growth factor (VEGF) producing human ovarian cancer cell lines. Finally, expression of FGF-2, phospho-Akt was assessed by Western blotting and flow cytometry. Results: We demonstrated the role of Akt-activated ECs in supporting expansion and self-renewal of ovarian cancer cells (OCC) in a residual disease context. We demonstrated that OCCs activate the endothelium, which displays resistance to bevacizumab. Bevacizumab had no effect on the proliferation of Akt-activated ECs, but significantly inhibited angiogenesis and delayed wound healing in HUVEC. We demonstrated that in this setting the cross-talk between cancer cells and ECs activates Pi3k/Akt inducing an autocrine loop through the pro-angiogenic factor fibroblast growth factor-2 (FGF-2) bypassing the VEGF-R pathway. We demonstrated that FGF-2 blocking would efficiently reverse the resistance to bevacizumab. Conclusion: Our data highlights the role of an activated endothelium in the constitution of the residual disease and resistance to bevacizumab. These results hint on the concept of using combination therapy to override drug resistance in ovarian cancer and to suppress or eradicate residual disease.

A consanguineous Arab family affected by a novel autosomal recessive disorder characterized by severe mental retardation and failure to thrive, was studied by Illumina 700K SNP genotyping, candidate gene mutation screening and whole exome sequencing for one affected member. Clinical findings include ptosis, bilateral epicanthic folds, striking midface hypoplasia, downturned mouth corners, thin upper vermillion, prominent ears, bilaterally short fourth metatarsal bone, bilateral fifth finger camptodactyly, mildly limited mobility in both knees and hypotonia. The gene was mapped by homozygosity mapping to 4 possible genome intervals, as the number of family members was insufficient for a significant LOD score. The positional candidate MAP2K1 was sequenced. No pathogenic mutations were identified. Whole exome target enrichment sequencing was performed on ABI SOLiD 4 System and Illumina HiSeq platforms for a single affected individual. Six non-synonymous variants were positioned within the homozygosity intervals. Two affecting evolutionary highly conserved amino acids with damaging effects according to PolyPhen and SIFT protein-modeling software, which were validated by Sanger sequencing to determine whether variants co-segregated with the disease phenotype. The variants were absent in 188 ethnically matched control chromosomes. One gene shows very limited expression in the brain while the other, a zinc finger protein, appears to be the disease causing mutation. Zinc finger proteins, a family of DNA and RNA binding proteins are transcriptional regulators controlling developmental cascades of gene expression especially during fetal brain development. Mutations in zinc finger domains interfere with normal brain development, are associated with non-syndromic X-linked mental retardation with impairments in adaptive behavior, and manifest during the developmental period causing severe mental retardation. The c.C5054G [p.S1685W] mutation affects 2 of the 3 ZNF407 isoforms, is located in the last third of the zinc finger domains and affects a serine residue in the alpha-helical part adjoining two zinc finger domains. This is thought to eliminate the functionality of downstream domains and interferes with expression of various genes under ZNF407 control during fetal brain development. Homozygosity mapping and whole exome sequencing of a single affected individual was the most effective, least labor intensive and most economical approach in identifying the mutation for this novel autosomal recessive zinc finger protein that causes a mental retardation syndrome.

Background & Objectives: Stein-Leventhal syndrome commonly known as polycystic ovary syndrome (PCOS) is the most common endocrinopathy among reproductive females. PCOS is a multisystem challenge causing not only gynecological issues such as irregular menstrual cycles, hyperandrogenism and infertility but also causing insulin resistance, type 2 diabetes and cardiovascular diseases. The objective of our study was to determine the incidence of PCOS among reproductive females at Qatar University using the National Institutes of Health (NIH), the Rotterdam consensus (Rott.) and the Androgen Excess Society (AES) criteria to determine the common phenotypes and hormonal parameters found in Qatar. Our aim was also to conduct a community-based study to make up for the lack of work done in female healthcare. Methods: A sample size of 121 females between the ages of 18-25 years were evaluated for symptoms of PCOS using family history of PCOS, body mass index and hormonal analysis. The sample was divided into the following four groups: control (n= 49), irregular menstrual cycles only (n= 13) , hyperandrogenism only (n= 27) and PCOS group (n= 31). Blood was drawn for measurement of TSH, progesterone, insulin, estradiol, SHBG, testosterone, DHEAS and prolactin. Modified Ferriman-Gallwey score was used for the evaluation of hirsutism. Results: The incidence of PCOS was found to be 25.83% (n= 31) using the Rotterdam criteria. Lower values were achieved using AES (20.83%, n= 25) and NIH (19.17%, n =23) definitions. Among our PCOS group, 16.13% (n= 5) were Qataris, 48.39% (n= 15) were Asians, 22.58% (n= 7) were Africans and 12.90% (n= 4) Europeans. Among the clinical features, family history of PCOS (p=0.037), hirsutism (p= 0) and irregular menstrual cycles (p= 0) were found to be significant factors in the PCOS group. Biochemical analysis showed elevated testosterone, elevated DHEAS and decreased SHBG hormones to be significant factors found in the PCOS group. Insulin and prolactin did not show any significant differences between the groups. Four individuals from the PCOS group were aware of their condition while 87% were unaware of their condition. Conclusions: Our study revealed a fairly significant prevalence of PCOS in the region and indicated a lack of awareness among people about their condition. There is need for government initiatives towards adequate investigations, management and treatment of PCOS.

Background: Apolipoprotein E (ApoE), a protein component of blood lipid particles, plays an important role in lipid transport and delivery. Single polymorphisms in residues 112 and 158 define the common E2, E3 and E4 alleles. In a study of Qataris, we observed that 17.4% of the African-derived genetic subgroup were heterozygotes for the rare Arg145Cys (R145C) variant that functions as a dominant trait with incomplete penetrance associated with dyslipidemia. Based on this, we hypothesized that the R145C polymorphism may be common in African-derived populations. Methods: The prevalence of the R145C variant worldwide was assessed in the 1000 Genomes Project (1000G) and then in 1012 Caucasians and 1226 African-Americans in New York City. Lipid profiles of the Qatari and New York R145C+ heterozygotes were compared to controls. Results: R145C+ Qatari heterozygotes had higher triglyceride levels compared to Qatari controls (p <0.007). The 1000Gs data demonstrated that the R145C polymorphism is rare in non-African derived populations, but present in 4.9-12.3% of Sub-Saharan African-derived populations. The R145C polymorphism was rare in New York City Caucasians (1/1012, 0.1%), but strikingly, 53 (4.3%) of 1226 New York City African-Americans were R145C+ heterozygotes, with an average of 52% higher fasting triglyceride levels compared to African-American R145C- controls (p <0.002). Conclusions: Based on these observations, there are likely to be millions worldwide derived from Sub-Saharan Africans that are ApoE R145C+. While larger epidemiological studies will be necessary to determine the long-term consequences of this polymorphism, the available evidence suggests it is a common cause of a mild triglyceride dyslipidemia.

Objectives: Stroke prevention therapy with oral anticoagulants (OAC) is reported to be under-utilized in patients with atrial fibrillation (AF) despite the overwhelming evidence supporting their use for stroke prevention. Rates and trends of the use of OAC from the developing world in this setting are lacking particularly in elderly patients. The aim of this study is to evaluate trends of utilization of OAC in elderly patients hospitalized with AF in a real-world population in a Middle-Eastern country over 20 years. Methods: Retrospective analysis of prospective registry of all patients aged above 70 years hospitalized with AF in Qatar from 1991 through 2010 was made. Rate and trends of OAC use was examined over the 20 years of the study. Results: During the 20-years period; 744 patients above the age of 70 years old were hospitalized for AF. Overall 26.1% of these patients received OAC with warfarin at hospital discharge while 60.2% received anti-platelets therapy with aspirin. The use of warfarin in this elderly group was significantly trending higher over the study period from 3.7% in the earlier years of the study to 39.5% in the latter years (P= 0.001). Anti-platelets therapy with aspirin also significantly increased from 47.4% in the earlier years of the study to 67.3% in the latter years (P= 0.001) [Table]. Conclusions: Although our study demonstrates that stroke prevention therapy with OAC and anti-platelets therapy are used with increasing frequency in elderly patients aged above 70 years over the 20-year study period, nonetheless they remain under-utilized. Further prospective studies are needed to investigate the reasons behind under-utilization of OAC in our region to help guide healthcare providers involved in the care of elderly people with AF to maximize potential benefit from OAC and anti-platelets while minimizing the hemorrhagic risk associated with these drugs.

Background: The incidence and prevalence of type 2 diabetes mellitus (T2DM) is on the rise in Qatar. The pathogenesis of T2DM is complex owing to molecular heterogeneity in the afflicted population. Current diagnostic methods rely on blood glucose measurements, which are non-informative with respect to progression of the disease to other associated pathologies. Thus predicting the risk and development of T2DM-related complications like cardiovascular disease remains a major challenge. Methods: We have used a combination of quantitative methods for characterization of circulating serum biomarkers of T2DM using a cohort of non-diabetic control subjects (n=76) and patients diagnosed with T2DM (n=106). In this case-control study, the samples were randomly divided as training and validation data sets. In the first step iTRAQ (isobaric tagging for relative and absolute quantification) based protein expression profiling was performed for identification of proteins displaying a significant differential expression in the two study groups. Five of these protein markers were selected for validation using multiple reaction-monitoring mass spectrometry (MRM-MS) and further confirmed with Western Blot and QRTPCR analysis. Results: We report the identification and verification of several biomarker candidates that can be potentially used for subset stratification of T2DM cohort. We identified a total of 227 high confidence proteins corresponding to 1393 peptides with greater than 95% confidence representing >40% sequence coverage. GO analysis predicted the differentially expressed proteins to be involved in a wide variety of cellular and metabolic processes. A total of 124 proteins showed a fold change of 2 or more between the control and T2DM serum samples. We report a set of five proteins viz. apolipoprotein-A-1, vitamin D binding protein, fibronectin, afamin and transthyretin as potent biomarkers, which are significantly up-regulated in diabetics. Conclusion: Our study revealed that many distinct molecular networks and metabolic pathways were activated in T2DM. It is hoped that future studies with larger cohorts as well those that focus on dissecting mechanistic roles will enhance our understanding of pathogenesis of T2DM and contribute to the development of targeted therapeutics in accordance with the personalized medicine paradigm.

Background and Objectives: Hematopoietic stem cells (HSC) are the most widely studied and characterized adult stem cells, which play an essential role in sustaining the formation of blood and immune system. The ease of their manipulation, the lack of serious ethical issues, and, in the autologous setting, the absence of their immunogenicity, have made them an attractive tool for developing stem cell-based therapies. Exploiting the properties of HSC by microRNA (miRNA) profiling offers an attractive approach to identify new regulators of stem cell fate. Although numerous miRNA have been screened from hematopoietic stem cells (HSC), the targets corresponding to many of these miRNA have not yet been fully elucidated. Therefore the objective of this study is to generate a miRNA profile from a subpopulation of adherent CD34+ HSC isolated from G- colony-stimulating factor mobilized peripheral blood aiming to understand the role of selected miRNA in regulating HSC stemness. Methods: CD34+ cells from patients' blood were isolated using a CD34+ isolation kit (Miltenyi Biotec) according to the manufacturer's protocol. miRNA profiling of adherent and nonadherent CD34+ cells was done using TaqMan Array MicroRNA Cards. Nanog expression levels was tested using a dual-luciferase reporter construct for miR-181a* or its mutant variant and Nanog 3′ UTR mRNA. Results: In this study, we have identified eight clusters of miRNA that were differentially expressed in an adherent subpopulation of CD34+ stem cells. Further analysis of one of the clusters by bioinformatics revealed that a miRNA, miR-181a*, which is highly expressed in the adherent CD34+ cells, affects the expression levels of Nanog, a stem cell surrogate marker. We show specifically by reporter assay and mutational analysis that miR-181a* targets a seedless 3′ compensatory site in the 3′UTR of Nanog and affects gene expression. We demonstrate that inhibiting miR-181a* upregulates the Nanog expression level, in addition to an increase in alkaline phosphatase activity. Conclusions: In conclusion, our results highlight a new stem cell-related target for the miR-181 family and show that miR-181a* directly targets Nanog in a subpopulation of CD34+ stem cells suggesting a possible role of miR-181a* in regulation of adherent CD34+ HSC cells stemness.

Background: Novel survey instrument development requires reliability testing, but limited information informs the preferred approach in Qatar. The testing of survey characteristics of the Multi-Cultural Assessment Inventory-Qatar (MAI-Q), a novel multicultural assessment tool in Arabic, English, Hindi and Urdu, provided the opportunity to investigate survey distribution procedures. Objective: To determine willingness to participate, and response rates during survey reliability testing by mail and in person. Methodology: To examine participation preferences and rates, patients were recruited through HMC and six primary health centers. When recruited, subjects could choose to take the MAI-Q a second time for the reliability testing in person or by mail. Subjects were either scheduled for follow-up in-person, or were sent the survey in a prepaid, preaddressed envelope. Results: Among 387 subjects who initially took the survey, 177 (46%) declined taking the survey a second time for reliability testing. Of 210 subjects who agreed to take the survey a second time for reliability testing, 145/210 (69%) chose mail, and 65/210 (31%) were offered and chose in person follow-up. Actual follow-up to complete reliability testing was only 29/145 (21%) by post but 40/65 (63%) in person. Conclusion: These findings reveal challenges researchers face when developing and distributing surveys in Qatar. Nearly half of subjects initially recruited were not willing to participate in follow-up testing. Among those agreeing to follow-up testing, the majority chose mail, but only a fifth of them returned instruments. The majority of subjects who chose in person follow up completed testing. These data inform researchers about the tough choices for developing and conducting surveys in Qatar.

Background: In order to monitor the health of new borns , high performance sensors are placed on the body to record information from different organs: nervous system (EEG signals), heartbeat and fetal movement (accelerometer signals). As the received data are non-stationary signals, Quadratic Time-frequency distributions (QTFDs) are often used to represent the energy, temporal and spectral characteristics of these signals. The suitable QTFD should conserve a high resolution and removes the cross-terms artifacts given by inner- and outer- terms. The inner-terms artifacts are observed in mono-component signals. Due to the Quadratic form of the QTFD, outer-terms are added to inner-terms if the signal contains more than one component. Recent QTFDs such as Wigner-Ville distribution, Spectrogram, B distribution, Modified B-Distribution, Choi-Williams, focus on the elimination of outer-terms whereas few works dealt with the inner-terms artifacts. Objectives: This work aims at designing a new kernel that takes into account the presence of both inner-terms and outer-terms artifacts. The proposed QTFD should remove these artifacts while maintaining a high resolution. Methods: A theoretical analysis of the localization of the inner-terms is given. Based on this analysis, we propose first a new kernel that removes the inner-terms for mono-component signal. Multicomponent signals are also considered, in which case we extend the proposed kernel in order to take into account the presence of the outer-terms. Results and conclusion: The resulting TFDs are compared with other methods using several examples of mono-component and multi-component signals. Some real data representing EEG signals and fetal movement are also used. The different simulations indicate that the proposed TFDs are more efficient to the existing ones, leading to potential significant improvement in newborn health outcomes. This work is supported by Qatar National Research Fund, NPRP project No 09-626-2-243

Interprofessional Education (IPE) is defined by the World Health Organization (WHO) as the process by which a group of practicing health-care professionals work together in order to provide promotive, preventive, curative, rehabilitative and other health-related services. WHO strongly encourages efforts to develop and integrate IPE into educational health-care programs for patients' benefits. For this to occur, a set of shared core competencies must be developed and used as a base on which to build and strengthen interprofessional skills in health-care professionals. Little or no research has been made to develop a set of IPE shared core competencies in Qatar. This article describes the method and results of the competency specification process developed by our team. At a first stage, a list of domains with their descriptions were developed using a card-sorting approach. A pyramid model with IPE at the top was also developed. The second stage was to develop the list of shared core competencies related to the domains developed above. The method used was a Delphi-based approach. One of the necessary steps was to draft candidate competencies based on 8 local experts and review of relevant literature. Based on this, feedback from local experts was integrated in an iterative process. The list of competencies was reduced from 20 statements approximately per domain to an average of 6 statements per domain. A rubric was also developed which can be used to evaluate students on the dimensions outlined in the shared core competencies.

Background & Objectives Large-scale proteome analysis by mass spectrometry is commonly preceded by enzymatic digestion of proteins. The conventional protocol for in-solution digest of complex protein mixtures includes trypsin and is performed at room temperature for at least 12h. To improve this time-consuming method we assessed the efficiency of focused ultrasound-assisted enzymatic digest. It has previously been shown that ultrasonication can dramatically reduce digestion time. However, it is not known how this affects protein identification and quantification when applied to complex protein mixtures such as whole cell lysates. Here, we explore in-depth the proteome of embryonic stem cells comparing trypsin-digestion methods with and without ultrasound support side-by-side. Methods In-solution protein digest Protein extracts were digested with sequencing-grade trypsin (Promega) for 1, 5, 10, 20, 30, 40min and 1h in the presence or absence of focused ultrasonication (Covaris). A conventional digestion overnight was included. Reactions were stopped by acidification and heat. Peptides were desalted by reverse phase C18 StageTips. Mass spectrometry Peptides were analyzed on a QExactive (Thermo Scientific) coupled to Easy nLC 1000 (Proxeon). Raw files were processed by MaxQuant v1.3.0.3 software. Peptide searches were performed by Andromeda search engine against the UniProt database. Results and Conclusions Focused ultrasonication clearly enhanced the enzymatic digestion and can be implemented in the daily workflow of mass spectrometry-based proteome analysis.

Introduction: Acute gastroenteritis is a widespread disease that is characterized by diarrhea, vomiting and abdominal pain. It has high morbidity and mortality rate globally. Bacteria, parasites and mostly viruses cause acute gastroenteritis. Most frequent viruses are norovirus, adenovirus, astrovirus and rotavirus. Objectives: To determine the prevalence of the four viruses in the stool samples of patients who were treated at HGH with complaints of acute gastroenteritis. The samples which had been submitted to the microbiology laboratory at HGH, were analyzed by real time-PCR method at Qatar University. The second objective was to compare between ELISA, IMCG and RT-PCR in terms of sensitivity and specificity. Methodology: 288 stool samples and corresponding patients' data were obtained from microbiology laboratory in HMC between 15-Jun-2009 to1-Nov-2009. They were transported to the research lab of biomedical science program- Health Department at Qatar University, where laboratory experiments involving RT-PCR were performed. Statistical analysis was carried out using Excel program. Results: Out of total 288 fecal samples, 131 (45.5%) were positive for the presence of viruses that cause gastroenteritis. Norovirus (28.5%) was the most prevalent, followed by rotavirus (10.4%), adenovirus (6.25%) and astrovirus (0.30%). Conclusion: Norovirus is the most common source of acute gastroenteritis among hospitalized patients, followed by rotavirus. Real time-PCR technique used in this study was found to be more sensitive and specific. By taking RT-PCR as a reference, ELISA and IMCG are found to be similarly low in sensitivity and specificity.

This work presents the development of an algorithm for analyzing ECG waveforms. The identification of the various waveforms on an ECG is the first and most crucial step in any automated analysis. The algorithm developed is capable of detecting all important waveforms. These include the Q,R,S,R' and S' waves, J and ST points and onset and offset of P and T waves. The various techniques utilized in their detection include adaptive thresholding, parabolic curve fitting, modified derivatives and temporal coherence. After detecting the waveforms, various measurements are obtained from the points detected. These include the polarity of the waveforms, the ST elevation, morphology of ST segment, QRS width and T wave morphologies. These measurements can easily be utilized for diagnosing the ECG for various abnormalities. The developed detection and measurement algorithm stands out from previous works in a number of ways. It is comprehensive and capable of detecting all the important ECG waveforms. It works well even on atypical ECG beats with secondary R' and S' waves. The adaptive thresholding approach minimizes the dependence on fixed hard thresholds. The synchronous behavior of ECG recordings across leads is exploited to improve accuracy. The performance of the algorithm was validated on 40 sample 12-lead ECG data of two minutes each. To validate performance on abnormal ECG waveforms, 20 datasets from patients diagnosed for myocardial infarction (MI) were included. An overall detection accuracy of 98.5% was obtained.

Animal models are essential to understand the molecular and cellular basis of human pathologies. The zebrafish has gained significant attention as a novel model system and has several advantages over rodent models, including: (1) genetic and physiological similarity to humans, (2) a high fertility rate, (3) external fertilization, (4) ease of genetic manipulation, (5) enlarging library of heart-specific reporter genes, (6) ability to survive with a dysfunctional heart, (7) transparency enabling powerful imaging modalities. The zebrafish heart has only two chambers but beats with a heart rate comparable to humans. Several genetic models have been engineered, which successfully model cardiac conditions. Random integration of reporter genes by transposon-mediated transgenesis generated a large number of zebrafish lines, specifically labeling different cardiac cell types and tissues. These include marker genes that label the myocardium, epicardium, endocardium, the conduction system, and atrioventricular and aortic valves (see Figure). Such reporter lines are useful to monitor specific cardiac cell populations and to study their role in disease processes. Similar efforts have recently been employed to identify the cell population responsible for cardiac regeneration. Gene mutations identified in humans causing cardiomyopathies have been introduced in zebrafish and successfully modeled the disease process in this species. Moreover, chemical genetic approaches have been successfully employed to identify small molecules, which are able to reverse pathological conditions such as hypertrophic cardiomyopathy. These examples demonstrate the power and versatility of this model organism, which will pave the way for novel therapeutic opportunities to treat cardiovascular disease. In the Qatar Cardiovascular Research Center we plan to utilize the zebrafish model to study the genetic basis of aortic valve disease, hypertrophic cardiomyopathy, and heart failure. A zebrafish lab will be established in Doha as part of the network in order to develop local expertise and to contribute to the overall research efforts.

Progression of colorectal cancer (CRC) culminates in liver metastasis. To identify genes that are involved in the metastatic phenotype, cDNA microarrays were used to analyze mRNA expression profiles of CC531 rat colon adenocarcinoma cells for changes related to their homing into the liver. Briefly, CC531 cells were intraportally implanted into the liver of Wag-Rij rats and re-isolated after 3, 6, 9, 14 and 21 days. Altogether 2919 genes from ca. 25000 genes were deregulated as a response to metastasis formation into the liver. Candidates from promising gene families were chosen for further analysis using different in vitro models and human CRC samples. These gene families included the claudins and the insulin-like growth factor binding proteins (IGFBPs). For claudins, it was shown that the initial phase of rat CRC cell homing into the liver involves a transient down-regulation of several claudins. Also it was shown that siRNA silencing of Cldn1 and Cldn4 increased migration and reduced colony formation, with these phenotypes being consistent with metastatic homing. Correspondingly, human CRC samples showed increased Cldn1 and Cldn4 expression in UICC stages l-lll, and significantly reduced expression in stage IV and in liver metastasis. Additionally, the IHC analysis revealed that the expression of Cldn1, Cldn4 and Cldn3 in liver metastasis specimens was lower than in the corresponding primary carcinomas. Taken together, metastasizing cells require a transient reduction in claudin expression for liberation from the primary tumor and then initialization metastatic growth in the liver. For IGFBPs it was shown that deregulation of IGFBP3 and IGFBP7 is related to the metastatic behavior of CRC cells. In CRC patients, a significant correlation between the expression levels of these genes was noticed but no relation to overall survival was registered. Further, the patients' age was inversely correlated with the expression of IGFBP3. Silencing of IGFBP3 and/or IGFBP7 in human and rat colorectal cancer cell lines reduced proliferation, colony formation, and for IGFBP3, also migration. These results indicate that IGFBP3 and 7 cannot be simply assigned to the group of tumor suppressors, but have additional properties, which become evident only in the context of cancer progression and metastasis formation.

Objectives: There is paucity of data relevant to anticoagulation rates and trends of treatment in patients with atrial fibrillation (AF) in developing countries and whether there are ethnic differences in the utilization of warfarin as some reports have suggested. The aim of this study is to evaluate trends of utilization of warfarin in Arab and South Asian patients hospitalized with AF in a real-world population in a Middle Eastern country. Methods: Retrospective analysis of prospective registry of all patients hospitalized with AF in Qatar from 1991 through 2010 was made. Rate and trends of warfarin use was examined in relation to ethnicity over the years of the study. Results: During the 20 year period; 2857 Arabs and 548 Asians were hospitalized for AF. The mean age of Arabs was 58 years and of Asians 49 years. Overall, 31.8% of Arabs and 39.4% of Asians received warfarin at hospital discharge (p= 0.001). The overall use of warfarin was significantly trending higher from 17 % in the earlier years of the study to 44.1% in the latter years but no differences were found between Arabs and Asians throughout the study period except at the latter years of the study when it was significantly higher in Asians (53% Asians vs. 41.7% Arabs; p= 0.009). Conclusions: Although the trend is significantly increasing over time, warfarin remains underutilized in patients hospitalized with AF irrespective of ethnicity. There is an urgent need for prospective studies to investigate warfarin contraindications, relative warfarin efficacy and bleeding risks in our region to help guide healthcare providers prescribing warfarin and consequently reduce the risk of AF-related disabling strokes.

Background: There is continuing concern about the risk of long-term autograft dilatation and rupture following the Ross procedure. The aim of this study is to characterize the neo-aortic valve in terms of morphology and wall distensibility. Methods: Among 108 Ross patients undergoing yearly echocardiographic assessment in our unit, 17 (16%) had aortic root diameters of >45mm (DG) 14 ± 1 years after operation. In an attempt to characterize the autograft morphology and dynamics, 11 patients (age 47 ± 10) with dilated autografts underwent cardiac multi-slice computed tomography (CT). Dimensions and distensibility of the aortic root were derived from the cine-CT images. Morphological observation was also made with rapid prototyping models based on the multi-slice CT. The patients were compared with 11 age-matched normal controls (C, age 47 ± 9). Results: Sinus diameters (Ds) were increased in DG compared to C (50 ± 3 mm vs 31 ± 4 mm, p < 0.01). The non-coronary sinus of the autograft tended to enlarge to a greater extent than other two sinuses. Aortic diameters (Da) were also increased in DG (35 ± 6 mm vs 28 ± 3 mm, p < 0.01). DG patients had similar cyclic strain (7.2 ± 3.8% vs 8.3 ± 3.6%, p = 0.13) and distensibility (25 ± 14 vs 34 ± 14 MPa-1, p = 0.20). Conclusions: These data support the notion that the autograft root has similar physical properties to the normal aortic root despite significant dilation. This may have implications for both the selection of aortic root substitutes and the clinical management of Ross patients presenting with dilated roots.

Collagen fibers are essential components of tissues and an important protein in the extracellular matrix, which maintains the structural and mechanical integrity of tissues while providing key signals to regulate cell functions. Although animal-based collagens can be used as biomaterial for tissue engineering heart valves, they cause infections and lack flexibility. These limitations have stimulated the exploration of collagen mimetic peptides (CMPs) through a bottom-up approach using computational modeling followed by experiments. Here, a 3D-structure of 3-helices of CMP was used to model the new CMPs to identify its structural stability and hotspots. These data assist in introducing charged residues by mutations to cross-link 3-helices and to add binding motif (GFOGER) for integrin in the CMP structures. The helical stability and self-association of the mutated CMPs have been validated using molecular dynamics (MD) simulations, surface electrostatic calculation followed by biochemical experiments using HPLC and mass spectrometry. The modelling analyses indicated that the CMPs show the desired structural properties for self-assembly and high affinity towards integrin binding. The introduction of the charged residues increased the possibilities for helical cross-link (gelation). The results suggest that altering the positions and length of hydrophobic repeats (GPO) of CMPs could improve thermal stability (Figure). The structural properties of the modelled CMPs were reproduced in experimental conditions. Our study provides potential peptide candidates that promise to show some inherent structural properties of native collagen in silico and in vitro. These properties are required to produce functional scaffolds for tissue engineering heart valves.

Our strategy for heart valve tissue engineering uses autologous cells to populate appropriate template matrices.The goal is to create a suitable scaffold supporting proper cell growth while reproducing the specific mechanical properties of native heart valves extracellular matrix (ECM). Accordingly, aligned and non-aligned nanofibrillar structures of different porosities were prepared and evaluated with regards to their mechanical properties and their ability to support human adipose derived stem cell (hADSC) colonization and growth. Nanofibrillar structures were obtained by jet-spraying poly (ε-caprolactone) dissolved in chloroform at different concentrations (0.0733, 0.0833 and 0.1033 mg/ml). Morphological evaluations of the structures were performed using scanning electron microscopy and porosity calculated. hADSCs (600,000) were top and rotary seeded on nanofibrillar discs (diameter 1 cm and thickness 0.8 mm) and cultured in 10 ml of complete medium under rotation (10 rpm) for 20 days. Histology (DAPI staining) and DNA quantification characterized the resulting cellularized structures. Jet-spraying polymers resulted in biphasic structures composed of both a dense and a nanofibrillar layer (Fig. 1-A). The nanofibers could either be of random orientation (isotropic) or well aligned (anisotropic). Isotropic matrices showed an increase in porosity when increasing polymer solution (96 to 97 %). Mechanical properties were inversely related to matrice porosities. Anisotropic matrices were stiffer than isotropic ones (elastic modulus up to 22 and 2 MPa respectively). hADSC proliferated significantly over 20 days within the scaffolds (up to 5-fold DNA increase), regardless of structure and porosity. The constructs dense polymer layer was favored by the cells up to 10 culture days, but after 20 days, the cells tended to bridge the entire scaffolds thicknesses (Fig. 1-B). This study highlights the potential of biphasic nanofibrillar matrices as substrate for valve engineering, particularly with regards to their elevated mechanical properties and capacity to support cellular population and proliferation.

Background & Objectives: School children among the most vulnerable groups that pass critical periods of physical, mental, and developmental growth are in need of special nutrition and health care. Little is known about the nutritional status of school children in Qatar. The aim of the present investigation is to determine the anthropometrics measurements (height, weight and body mass index) of Qatari school children, comparing the study results with the international reference population and to determine the relationship between obesity and lifestyle. Methods: Study was conducted on a sample from Qatari school children. A representative sample (1500 children) were selected between the ages of 6-12 from 23 schools using multistage cluster random sample, while 200 cases from each age group were targeted as primary sampling unit. Data about number of physical education classes, hours of physical activity per week, and information about the sedentary life style were obtained by utilizing a standardized self-administered questionnaire. Results: The current results showed that the percentage of body weight for Qatari children were significantly higher than the reference values while their body height percentage were significantly lower than the reference. Using the WHO (2007) classification of BMI for children, 8%, 16% and 23% of studied children had thinness, overweight and obesity, respectively. The majority of children (55.1%) spent more than 2 hours daily watching TV, surfing the internet or playing electronic games, 25.8% didn't report any kind of physical activity and 48.8% had physical activity less than one hour/day. The current study showed that there is a significant relationship between children's physical activity and prevalence of overweight and obesity. Conclusions: The current work looks at school children in Qatar to create a more complete picture of the current state of health and help to determine how lifestyle and environmental changes can be mitigated as a causal factor of obesity and therefore lifelong poor health.

Objectives: Although predictors of mortality have been identified in major atrial fibrillation (AF) clinical trials, most of the data were limited to studies in the developed world and included mainly Caucasian patients. The aim of this study is to identify predictors of mortality in Middle Eastern Arab and South Asian patients hospitalized with AF in a real-world population in a Middle Eastern country. Methods: Retrospective analysis of prospective registry of all patients hospitalized with AF in Qatar from 1991 through 2010 was made. Ethnicity, clinical characteristics and co-morbidities were analyzed in relation to mortality. Results: During the 20 year period; 2857 Arabs and 548 Asians were hospitalized for AF. There were 138 deaths among Arabs and 14 among Asians (4.8% versus 2.6%, p= 0.02). Multivariate analysis of predictors of in-hospital mortality identified chronic renal impairment and acute coronary syndromes as independent predictors of poor outcome among patients hospitalized with AF whereas ethnicity was not [Table]. Conclusions: In this region, the risk of in-hospital mortality for patients hospitalized with AF remains increased in patients who have chronic renal impairment or acute coronary syndromes irrespective of ethnicity. Patients at high risk may benefit from aggressive monitoring and intervention in order to reduce mortality.

Introduction: Obesity is a global problem affecting adolescents and teens with profound psychological and medical sequelae. The role of bariatric surgery in adolescents remains controversial. The aim of the study is to report our results with sleeve gastrectomy in this population. Goal of study: To study the efficacy of sleeve gastrectomy as a treatment for obese adolescents in terms of weight loss, comorbidity resolution and safety of procedure. Methods: Retrospective review of our records of all patients under the age of 19 who underwent laparoscopic sleeve gastrectomy in Hamad General Hospital in the years 2010-2012. Variables studied included; age, BMI, preoperative and postoperative weight, sex, comorbidities, length of stay, operative time, and early or late onset complications. Results: Out of 700 patients from our institute, 37 patients (5%) were adolescents. 45% males N=17 and 55% females N=20. Mean age was 17 ±1.5. Mean length of stay was 3.6 ±0.8. Mean operative time was 64 ±17 minute. Mean preoperative weight 141 ±27 kgs (200-94.) Mean BMI 51±8.8 kg/m2 (72-35). With regard to comorbidities, one person had obstructive sleep apnea and reported resolution. Mean follow up was 8 months (17-1), mean post op BMI 38 ±11.8 kg/m2 (23-70), mean excess weight loss -28% (4-45%). Mortality 0%. Morbidity 0%. Conclusion: Laparoscopic sleeve gastrectomy is a safe option for treatment of obese adolescents. It is particularly effective in this particular group with satisfactory safety profile and adequate weight loss. However, longer follow up is needed to evaluate the long term effects of the surgery.

Objectives: Little is known about the ethnic variability in trends of the etiologies of atrial fibrillation (AF) and related mortality. This was examined in this study in patients hospitalized with AF in a real-world population in a Middle Eastern country. Methods: Retrospective analysis of prospective registry of all patients hospitalized with AF in Qatar from 1991 through 2010 was made. Rates and trends of clinical characteristics, cardiac comorbidities and in-hospital mortality were analyzed according to ethnicity. Results: During the 20-year period; 2857 Arabs and 548 Asians were hospitalized for AF. Arabs were 9 years older and more likely to have hypertension, diabetes mellitus, chronic renal impairment and dyslipidemia compared to Asians. Valvular heart disease and acute coronary syndromes were more common among Asians while congestive heart failure was more common in Arabs. Overall in-hospital mortality was lower for Asians compared to Arabs while stroke rates were comparable [Table1]. There was an increase in the prevalence of diabetes mellitus and hypertension in both groups in the latter years of the study period but, more so in Arabs when compared to patients hospitalized in the earlier years. Associated valvular heart disease was trending lower in both groups while ischemic heart disease prevalence was trending higher with no significant differences between the two groups. Rheumatic heart disease was statistically trending lower in prevalence; however it remained more prevalent among Asian patients over the study period. There were no significant differences in mortality trends over the study period between the two groups [Table 2]. Conclusions: The current study is the first ever report of comparative ethnic trends in etiologies and outcome of AF among Middle Eastern Arab and South Asian patients over a 20-year period. The current study underscores the need to study differences in atrial fibrillation among various ethnicities.

Background and Objectives: Most previous explanations of fatigue during repeated sprint exercise have focused on muscular factors associated with cellular mechanisms. At present, the nature of the neural adjustments and the extent to which corticospinal responsiveness is altered after the completion of repeated sprint exercise are yet to be elucidated. This study was undertaken to test the hypothesis that, in addition to the large peripheral fatigue, supraspinal factors would contribute substantially to neuromuscular adjustments of the quadriceps muscle in response to a repeated sprint cycling exercise. Methods: Twelve active individuals performed 10 x 6-s 'all out' sprints on a cycle ergometer (recovery= 30 s), followed 6 min latter by 5 x 6-s sprints (recovery= 30 s). Surface electromyographic activity was continuously recorded during the exercise. In addition, transcranial magnetic and electrical femoral nerve stimulations during brief (5-s) and sustained (30-s) isometric contractions of the knee extensors were performed before and 3 min post-exercise. Results: Peak power output and vastus lateralis root mean square activity decreased across sprint repetitions (P <0.001). These decrements were significantly (r= 0.89; P <0.001) correlated. Maximal strength of the knee extensors decreased during brief and sustained contractions (∼11% and 9%, respectively; P <0.001). Cortical voluntary activation values (∼90%), motor evoked potential amplitude and silent period duration responses measured during briefs contractions were unaltered. While cortical voluntary activation declined (P <0.01) during the sustained maximal contraction in both test sessions, larger reductions occurred (P <0.05) after exercise. Lastly, resting twitch amplitude in response to both femoral nerve and cortical stimulation was largely reduced (>40%, P <0.001) following exercise. Conclusion: The novel findings provided by motor cortex stimulation are that exercise induced reductions in cortical voluntary activation were seen in sustained, but not brief, maximal isometric contractions of the knee extensors. This, along with the lack of corticospinal responsiveness impairment from pre- to post-exercise, indicates that supraspinal processes do not necessarily limit repeated sprint ability. Rather, fatigue exhibited by the quadriceps when cycling sprints are repeated resides within the muscle itself, due to limitations in energy supply and/or the intramuscular accumulation of metabolic by-products.

Background and Objectives: The provision of emergency care is globally recognized as among the most difficult and challenging elements of any healthcare system. Hamad Medical Corporation (HMC) was facing increasing pressure to introduce initiatives to significantly ease problems with congestion at Hamad General Hospital's (HGH) Emergency Department (ED). This ED is one of the busiest in the world, with an average daily census of 1,500 patients. Due to the large number of patients, fragmented design and inappropriate placement and distribution of resources, all previous attempts at decongesting the ED and improving patient flow were unsuccessful. HGH leaders knew from internal data that the large volume of the male low-acuity patients represented approximately 70% of total ED patients, and were the main reason for congestion and crowding. Therefore, targeting this group would bring about the most significant impact. Methods: Data were studied in order to predict the number of patients who would fit the low-acuity criteria. This study confirmed that up to 70% of all ED patients fell within the low-acuity criteria. Lean process mapping was conducted to determine the current state of patient flow, processes, and practices in the male low-acuity areas of the ED. The impact of the existing fragmented facility design was also studied. Detailed analysis of a patient's journey from arrival to disposition was undertaken. Results: The process mapping confirmed many unacceptable variations when managing low-acuity cases in the ED and it was evident new design and processes were required. A 'See and Treat' service was opened on 7 March 2010 resulting in: reduction in door-to-door transit time, reduction in adverse events and complaints, improvements in maintaining patients' privacy and confidentiality, more appropriate resource utilization, routine referring orthopedic team, better pathway for x-ray, pharmacy and cashier, and new documentation to support efficiency. Conclusions: We believe that the remarkable success of the 'See and Treat' service in such a challenging and unique department as ours is a clear evidence that it should be acknowledged as a benchmark for best practice for the provision of efficient and safe patient care in an emergency department, particularly when managing high-volume, low-acuity presentations.

Background and Objectives: There is an escalating interest among patients and consumers in the use of complementary and alternative medicines (CAM), specifically natural health products, for a variety of illnesses. The public interest in CAM makes it important to explore the attitudes and beliefs of pharmacists involved in selling and dispensing these products. Our objectives were to explore the attitudes of community pharmacists towards CAM and to gauge their opinions about the challenges faced when dispensing CAM during their daily practice. Methods: A qualitative exploratory study was conducted in Doha, Qatar. The Study utilized focus group meetings and a purposive sample of community pharmacists whose pharmacies sell CAM. Recruiting for focus group meetings continued until a point of saturation was reached. The discussions and responses to questions were audio-taped and transcribed verbatim. All recordings were analyzed for recurring themes and subsequently coded and categorized. Results: Most pharmacists felt they were not equipped with sufficient information to competently recommend CAM to their clients and all have no access to information resources to update their knowledge. Pharmacists who graduated from the Indian subcontinent shared strong belief in traditional Indian remedies which shaped their practice in respect to CAM. Pharmacists have doubts about the safety of CAM and felt their undergraduate pharmacy courses did not prepare them to adequately assist their patients. Conclusion: Pharmacists were not confident in dispensing or recommending CAM to their patients. In view of the escalating interest in CAM, and in order for pharmacists to support patient autonomy in selecting health care interventions, colleges of pharmacy should make radical changes in those undergraduate courses that teach subjects related to CAM.

Background and Objectives: Neonatal mortality, a component of United Nations Millennium Development Goal 4 (MDG-4) (two thirds reduction in childhood mortality by 2015), has been a focus of all global health improvement strategies and plans of action since 1990. Our study aims to prospectively ascertain Qatar's neonatal mortality rate (NMR) during 2011, analyze trends between 1975 and 2011, and compare with recent data from high income countries. Methods: A PEARL study (perinatal neonatal outcomes research study in the Arabian Gulf), a joint collaborative research project between Hamad Medical Corporation (HMC), Qatar and University of Gloucestershire, United Kingdom, is Qatar's prospective national perinatal epidemiologic study funded by Qatar National Research Fund. The study quantifies maternal, neonatal and perinatal mortality, morbidities and their correlates by establishing a national neonatal perinatal registry for Qatar called Q-Peri-Reg. Data on live births and neonatal mortality was collected from all public and private maternity facilities in Qatar during 2011 and compared with historical neonatal mortality data (1975-2011) ascertained from the database of maternity and neonatal units of the Women's Hospital and annual reports of HMC. Inter-country comparisons were made using World Health statistics 2011 and the European perinatal health report 2008. Results: Total live births during the study period were 20,583 and neonatal deaths 102. NMR was 4.9, early neonatal mortality rate (ENMR) 2.7, late neonatal mortality rate (LNMR) 2.2 and corrected neonatal mortality rate (cNMR) 3.26. Between 1975 and 2011, relative risk of NMR decreased by 87% (RR 0.13, 95% CI 0.10-0.18, p <0.001), ENMR by 91% (RR 0.09, 95% CI 0.06-0.12, p <0.001) and LNMR by 58% (RR 0.42, 95% CI 0.23-0.74, p= 0.002) though the population increased by 10 fold and number of deliveries by 7.2 folds. The comparable NMR from selected high income countries are between 3 and 5. Conclusion: Qatar has achieved its target MDG-4 by 2011. The improvement has been more marked in ENMR than LNMR. The current NMR, ENMR, LNMR and cNMR are comparable with selected high income counties. Further in-depth analysis of correlates and determinants of neonatal survival in Qatar may form the basis of a strategic global neonatal mortality improvement plan.

Background: Coronary artery disease remains one of the leading causes of morbidity and mortality in developed countries and is projected to be the leading cause of death in the developing world by 2010. Affecting nearly 12 million people in the USA, it accounts for 50% of all cardiovascular deaths and is the leading cause of congestive heart failure as well as premature permanent disability in workers. About 1.1 million myocardial infarctions and >400,000 new cases of congestive heart failure each year, cardiovascular disease severely impacts men and women as well as various ethnic groups. Objectives: The aim of this study is to evaluate the results of transmyocardial laser revascularization (TMLR) in patients with coronary artery disease, in whom percutaneous transluminal coronary angiography (PTCA) or coronary artery bypass grafting (CABG) cannot be done. Method: Fifty patients were included in the study: 1) Canadian cardiovascular angina class (CCS) III or IV patients (only 5 patients with CCS class II were included and those patients were the early trial in year 1997 where the recommendations for TMLR was not yet established); 2) Ejection fraction of 30%; 3) Those with evidence of reversible ischemia (based on stress thallium perfusion scanning); 4) Candidates of neither CABG or PTCA (according to the results of coronary angiography). The laser system used was a heart laser system (PLC medical systems). Results: There was a significant improvement in the severity of angina as assessed by Canadian Cardiovascular Society grading after the procedure. Improvement was noticed after 1 month and the improvement was most significant after 3 months and then slightly declined after 6 months but still significant in comparison to the pre-procedural values (P <0.05). Also, there was a significant improvement in the grade of dyspnoea as assessed by the New York Heart Association functional dyspnoea class and the most significant improvement was after 3 months and the degree of improvement decreased but still significantly better than the pre-procedural values (P <0.05). Conclusion: TMLR offers consistent amelioration of severe angina in patients having no conventional therapeutic alternative. Surgeons should recognize that the procedure is intended only for the purpose of reducing angina symptoms.

Background and Objectives: Type 2 diabetes mellitus (T2DM) is a major health problem in Qatar. The prevalence of T2DM in the adult population of Qatar is approximately 16% (twice that in the USA) and the country is ranked sixth in the world for diabetes prevalence. The number of cases of childhood diabetes in Qatar has doubled in ten years. The rapid rise of T2DM cases is thought to be due, in part, to genetic factors that interact with lifestyle factors including, but not limited to, poor nutrition, lack of physical activity and obesity. Recent evidence suggests that lack of vitamin D and inflammation may also play a role. Methods: We examined the current knowledge from literature linking T2DM to the chronic low-grade inflammation and the possible role of vitamin D in attenuating the inflammatory factors and, thereby, preventing T2DM. Results: Low-grade inflammation resulting from imbalances in the innate immune system has been associated with an array of chronic disorders that predispose to the later development of T2DM (e.g., obesity, metabolic syndrome, and insulin resistance). As a result, inflammation may contribute to the pathogenesis of T2DM. Therefore, attenuation of this inflammatory response via modulating the innate immune system could lead to improved insulin sensitivity and delayed disease onset. Dietary supplementation with vitamin D, may present a novel strategy toward the prevention and control of T2DM at the population level due to its anti-inflammatory and antioxidant properties. Conclusion: The concept that vitamin D, via attenuating inflammation, could be employed as a novel preventative measure for T2DM, is evaluated in the context of its relevance to healthcare and public health practices. Overall, to reduce the disease risk and burden at the population level, preventative strategies can be developed to include vitamin D supplementation into the traditional intervention protocols (e.g., exercise, life-style modification, etc.).

Background & Objectives: A recent randomized clinical trial, CAPRISA 004, reported levels of HIV incidence rate among general population women in South Africa that are comparable or higher than those observed among high-risk populations such as female sex workers, men who have sex with men, or injecting drug users despite low coital frequency. Understanding the factors leading to such high HIV incidence levels is critical for guiding prevention efforts. To this end, we have assessed the risk of HIV transmission among stable couples across a range of epidemic settings in Sub-Saharan Africa (SSA). Methods: We constructed a mathematical model describing the process of HIV discordancy in a population in the context of an HIV epidemic to estimate the risk of HIV transmission within stable partnerships. The model was parameterized using nationally representative demographic health survey data for 20 countries in SSA. Uncertainty and sensitivity analyses were performed to explore the robustness of our estimates to systematic biases in model parameters. Results: HIV incidence rate among discordant couples hovered around 10 per 100 person-years across SSA. A clustering based on HIV prevalence was observed with a median of 7.8 per 100 person-years in low HIV prevalence countries (<5%) compared to 18.4 per 100 person-years in high prevalence countries (>5%). HIV population prevalence explained 49% of the variation in HIV transmission, with every 1% increase in HIV prevalence implying an increase of 1.2 per 100 person-years in HIV infectiousness. The uncertainty and sensitivity analyses suggested that potential systematic biases are more likely to lead to a slight overestimation in HIV incidence rate particularly in high prevalence countries. Conclusions: Our results suggest that a variability in HIV infectiousness may have contributed to the contrasting HIV epidemic trajectories across Africa. The key drivers of this variability may include male circumcision, different circulating virus subtypes, sexually transmitted co-infections, tropical co-infections, hormonal contraception, and host genetics and immunology. The role of behavioral cofactors such as coital frequency and the uptake of condom use are less evident.

Background: Myeloproliferative neoplasms (MPNs), that is, essential thrombocythemia (ET), polycythemia vera (PV), and myelofibrosis (MF), are a group of MPNs that can lead to significant rates of morbidity and mortality among affected patients. Symptomatic burden in MPNs is present in most of MPN patients. We sought to use the broadly applicable instrument MPN symptom assessment form (MPN-SAF) to assess symptoms in MF, ET and PV among populations of Qatar. Methods: Using the MF-SAF as a base instrument, we added several key additional symptoms previously identified as present in all subtypes of MPNs including headaches, concentration, dizziness, insomnia, sexual problems and mood changes on a 0 (absent) to 10 (worst-imaginable) scale. The MPN-SAF was administered jointly with the EORTC-QLQ-C30 as the co-validation instrument using prospective cohorts in Qatar. Results: MPN-SAF patient data: 123 MPN-SAF surveys were administered (English (45%), Arabic (55%) in 45 ET patients (36.5%); 35 PV patients (28.5%); 15 MF patients (12.2%); 28 MPN unclassified (22.7%); an average of 7.8 years (range 0 - 43 years)) from their MPN diagnosis. Participants were of age range (26 - 58 years) and gender (52% female) characteristic of disease. Prior hemorrhage (10%) and thrombosis (25%) were frequent. 78% of the patients currently receive cytoreductive therapy and 87% received cytoreductive therapy in the past. Patients and Symptomatic Burden: 19 items assessed in the MPN-SAF demonstrated consistently that the most common symptoms were decreased quality of life (93%), fatigue (84%) and insomnia (65%). The least common symptoms (<50% prevalence) were fevers (15%) and weight loss (10%), although symptoms are present in all 3 MPN subgroups. The majority found the MPN-SAF easy to understand (92%) and "addressed most of my MPN symptoms" (95%). Comparison to EORTC-QLQ-C30: Strong correlations existed between individual items represented on both the MPN-SAF and the EORTC-QLQC30 including pain, fatigue, appetite and insomnia (all p <0.001). Symptoms characteristic of advanced disease including weight loss, fever, and cough displayed lower Pearson correlations (r= 0.46, -0.08, and 0.38 respectively). Conclusions: The MPN-SAF is a comprehensive and reliable instrument which is available in multiple languages (including Arabic and English) to evaluate MPN-associated symptoms.

Background: Osteoarthritis (OA) is the most common joint disease in middle aged and older people. Despite the multiple modalities of treatment, the outcome is still poor and focuses on temporary measures to alleviate the symptoms. Objectives: We want to determine whether or not an intra-articular injection of a single dose of chondrogenic induced bone marrow mesenchymal stem cells (BMSC) can promote cartilage regeneration in surgically induced osteoarthritis in sheep. Methods: Sheep BMSCs were isolated and divided into two groups. One group was cultured in chondrogenic media containing 5 ng/ml TGFβ3 + 50 ng/ml IGF-1, and the other group was cultured in basal non-chondrogenic media for the duration of 3 weeks. The procedure for surgically inducing osteoarthritis was performed on the donor sheep six weeks prior to intra-articular injection to the knee joint. The injection is a single dose of BMSCs from either group, suspended in 5 ml Dulbecco's Modified Eagle Medium (DMEM) at density of 2 million cells/ml. The control groups were injected with basic cell free media. Results: Six weeks after injection, evidence of articular cartilage regeneration and meniscus repair in osteoarthritic knee joints treated with autologous BMSCs cultured in chondrogenic medium were observed. No evidence of regeneration and meniscus repair was observed for the control group and the group treated with BMSCs cultured in basal medium. Conclusions: Intra-articular injection of a single dose of BMSCs in chondrogenic culture could stimulate regeneration of articular cartilage and meniscal tissue.

Background and Objectives: A major challenge to global health initiatives is food safety. In the USA it is estimated that 76 million foodborne illnesses occur each year causing 300,000 hospitalizations and 5,000 deaths. Escherichia coli O157:H7, Campylobacter and Salmonella enterica are among the leading causes of these illnesses including inflammatory bowel disease (IBD) with severe sequelae. The severity of illness is in part attributable to the virulent factors encoded by genes present in individual pathogens. Data on the risk associated with infection by foodborne bacterial pathogens in Qatar and the complement of virulent factors in these pathogens is lacking. Cost-effective intervention strategies rely on accurate microbial risk assessment (MRA) of the adverse consequences of exposure and infection with specific pathogens (dose-response relationships), but also according to the complement of virulent factors present in each agent. We carried out molecular studies to investigate the profile of virulent genes of three major foodborne bacterial pathogens isolated from humans in Qatar in support of developing accurate MRA. Methods: Multiplex polymerase chain reactions (PCR) test and PCR restriction fragment length polymorphism were used to examine E. coli O157:H7 isolates for virulent factors. PCR amplification was used to screen C. jejuni and C. coli for the presence of cytolethal distending toxin (cdt) subtypes (cdtA, cdtB, and cdtC). Results: E. coli isolates had a number of virulent factors in common. The enterotoxigenic E. coli (ETEC) pathotypes were detected in human samples at a rate of 40%, while Shiga-toxin producing E. coli (STEC) pathotype was detected at a much lesser rate. The attaching and effacing E. coli (AEEC) pathotype which included several serotypes (O26, O45, O103, O111, O121 and O145) causing illnesses in 2010 were detected at a rate of 60%. The cdtB gene was detected in 82% and 30% of the C. jejuni and C. coli isolates, respectively. The majority of Salmonella enterica isolates expressed cdtB gene. Conclusions: The data indicate that infection by foodborne bacterial pathogens are common causes of gastroenteritis in Qatar. Implementation of cost-effective control measures for these important foodborne pathogens require intervention strategies that take into account MRA within the context of Qatar.

Background: Anterior cruciate ligament (ACL) tears are common injuries occurring upwards of 250,000 times annually only in the US. These injuries create an annual billion dollar expense in the US alone. It is important to understand these mechanisms of injury as these injuries continue to be one of the largest problems in orthopedic sports medicine. Improved understanding of the injury mechanisms may improve prevention, rehabilitation and surgical procedures. Objectives: To perform a parametric study of knee joint movement to highlight which movement combinations are most detrimental to the ligaments of the knee joint, and also to study the effect of ramping rate on varus and valgus failure angle. The study provides a spectrum of injury propensity based on internal and external femoral rotation and varus or valgus angle as well as showing the effect of ramping rate on the prescribed boundary conditions. Methods: Bones and ligaments were digitized from magnetic resonance images (MRI). Bone was modeled as rigid, and a transversely isotropic visco-elastic material was applied to the ligament structures. This study incorporates a novel approach for developing bundle specific prestrain within 3D ligament structures. The bundles were stretched from their zero load lengths to their reference lengths, producing a strain field mimicking the in vivo strain conditions of the ACL at full knee extension. A failure locus was created by performing multiple FE simulations of knee joint motion combinations until ACL failure at different ramping rates to investigate the effect of rate dependency on the valgus and varus failure angle of the ACL. Results: Loading rate had negligible effect on the valgus or varus ACL failure angle. External femoral rotation scenarios were more susceptible to ligament failure than internal femoral rotation. The postero-lateral bundle showed more injury susceptibility compared to the anteromedial bundle. Varus simulations showed less angle needed for failure when compared to valgus simulations. Conclusions: These results have various clinical applications. In sports where ACL injuries are prevalent, training programs can be adapted to address the avoidance of harmful knee orientations.

Background and Objectives: The State of Qatar has achieved maternal, neonatal and perinatal survival rates which are comparable to many high income countries, both from the West and East. Our study aims to analyze sociodemographic, economic and cultural determinants of Qatar's neonatal mortality rate (NMR) during 2011. Methodology: PEARL study (Perinatal Neonatal Outcomes Research Study in the Arabian Gulf), a joint collaborative research project between Hamad Medical Corporation (HMC), Qatar, and University of Gloucestershire, United Kingdom, is Qatar's prospective national perinatal epidemiological study funded by Qatar National Research Fund. The study is quantifying maternal, neonatal and perinatal mortality, morbidities and their correlates by establishing a national neonatal perinatal registry for Qatar called Q-Peri-Reg. Data on live births and neonatal mortality was collected from all public and private maternity facilities in Qatar during 2011. Data on sociodemographic, economic and cultural determinants was ascertained by direct interviews from the mothers using predesigned performas. Univariate and multivariate regression analysis was done using Epi Info and SPSS-20. Results: Qatar's NMR during 2011 was 4.9. Babies born to mothers with no or less than secondary level of education were twice at risk of death as compared to mothers with secondary or more level of education (RR 2.08, 95% CI 1.23-3.53, p= 0.009). The rate of maternal literacy was 90%. Nationality, gender, consanguinity, maternal occupation, family income and status, type and ownership of housing did not have any statistically significant correlation with the risk of neonatal mortality (Table1). Conclusion: High level of maternal literacy in Qatar appears to have contributed significantly to its improved neonatal survival rates. The outcome is independent of nationality, gender, social, economic and living status which supports the notion of an equitable health system in Qatar.

Background and Objectives: Vitamin D is thought to modulate cardiometabolic disease risk through effects on an array of disease-associated pathways, such as glucose and lipid metabolism and inflammation. Although vitamin D insufficiency is thought to play a role in the susceptibility to cardiometabolic disease, the association between the two conditions is yet to be investigated in both men and women from the general population. Methods: Subjects (n= 1,928, age range 16-79 yrs) from Canada were examined. Plasma 25-hydroxyvitamin D [25(OH)D] by month of clinic visit were plotted for men and women to examine the monthly fluctuation patterns. Linear regression models adjusted for sex, waist circumference, physical activity, hormone use, and season of clinic visit examined the linear associations between 25(OH)D and cardiometabolic biomarkers [glucose, insulin, HOMA-IR, triglycerides, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total:HDL-C ratio, apolipoprotein(apo)A1, apoB] and biomarkers of inflammation [C reactive protein (CRP), fibrinogen, and homocysteine]. Analyses were repeated separately for men and women. Results: Monthly variation in 25(OH)D was noted for both men and women, with mean levels fluctuating more widely throughout the year in men (50 to 80 nmol/L) than in women (60 to 75 nmol/L). Significant inverse associations with 25(OH)D were noted for insulin (β= -0.16, p=0 .003), HOMA-IR (β= -0.18, p= 0.002), triglycerides (β= -0.21, p= 0.004), total cholesterol (β= -0.31, p= 0.046), LDL-C (β= 0.09, p= 0.02), and total:HDL-C ratio (β= -0.09, p= 0.004). When the population was stratified by sex, significant inverse associations with 25(OH)D were observed for insulin (β= -0.21, p= 0.002), HOMA-IR (β= -0.24, p= 0.002), and triglycerides (β= -0.26, p= 0.01) among men, and insulin (β= 0.13, p= 0.05), HOMA-IR (β= -0.14, p= 0.03), total cholesterol (β= -0.34, p= 0.05), LDL-C (β= -0.10, p= 0.02), and total:HDL-C ratio (β= -0.10, p= 0.001) among women. Conclusions: Plasma 25(OH)D levels vary month-to-month, and fluctuate more widely in men than in women. Vitamin D may modulate multiple processes related to the cardiometabolic biomarkers and disease risk and may, thereby, play a role in disease prevention.

Background and Objectives: Globally around half of new HIV infections occur in the age group 15-24. New reported cases of HIV/AIDS in Lebanon show that a younger age group (20-30) is being affected more than ever before. Lebanon is moving forward in implementing a national sexuality education curriculum in all public schools, as a mechanism for raising awareness and for preventing sexually transmitted infections (STI) including HIV among adolescents. However, little is known about adolescents' actual sexual behavior, the extent to which students are confident of their refusal skills, and the factors that determine these. Young adolescents, who have limited sexual knowledge and experience, may have difficulty in adopting the appropriate protective behavioral responses to prevent negative health outcomes including STI and HIV infection. Methods: This research identified the prevalence and risk factors for perceived sex refusal skills among Lebanese adolescents in grades 7-9 in public and private schools using the nationally representative Lebanon Global School Health Survey. Results: Results indicated that more than 40% (50.4% of boys and 34.8% of girls) of students did not feel confident that they could tell someone they did not want to have sexual intercourse with them. Multivariate logistic regression analysis indicated that being female, older and having close parental supervision predicted having perceived sex refusal skills among Lebanese adolescents. Moreover, having knowledge and/or education about risky behaviors was the strongest predictor for having perceived sex refusal skills among Lebanese adolescents. Conclusion: This study contributes to the limited literature on perceived sex refusal skills in Lebanon, the Middle East and North Africa. A number of recommendations for research and prevention programs can be drawn from this research. Advocacy efforts should target policy makers to ensure that policies are reviewed to support comprehensive sexuality education in Lebanese schools because of identified vulnerabilities of sexual violence, low levels of education or knowledge of risky behaviors including HIV risk, and low levels of perceived sex refusal skills.

Background and Objectives: Posaconazole (PSZ), is an antifungal prophylactic therapy that is used in hematologic cancer patients. It is approved for prophylaxis in hematologic cancer patients ≥13 years in USA, Canada, Australia, and ≥18 years in the European Union. In 2010, PSZ was added to the formulary of Al-Amal Hospital, the only adult cancer hospital in Qatar. The objective of this study is to conduct a drug use evaluation (DUE) study of posaconazole at Al-Amal Hospital. Methods: A retrospective, single centered, observational DUE study was conducted to include a convenient sample of hematologic cancer patients who used PSZ prophylactically during the year 2010. All patients, thirty-one, who received PSZ in 2010 were nominated of which 20 patients' profiles were reviewed, data were collected into a pre-prepared collection sheet and descriptive analysis was performed. Results: In Qatar, PSZ was used prophylactically in hematologic cancer patients >15 years with febrile and afebrile neutropenia. It is also planned to be used prophylactically for the upcoming bone marrow transplantation unit patients. All the 20 patients, 17 males and 3 females, received the PSZ for prophylaxis and were compliant. More than 50% of patients received proton pump inhibitors concurrently with posaconazole. Only 1 case had a recorded recommendation regarding the administration of PSZ with food. Five patients received vincristine-based chemotherapy protocol, one of which received it concurrently with PSZ for two cycles and developed seizure. Two patients developed mild breakthrough fungal infection, oral thrush, while on PSZ prophylactically. Conclusion: The PSZ regulations in Qatar are similar to the worldwide recommendations. The PSZ practice in Al-Amal hospital abides by the regulations. However, three points need to be addressed: PSZ has low bioavailability that can be enhanced by taking it with meals and by dividing the total daily dose. PSZ co-administration with proton pump inhibitors should be stopped as it may result in PSZ sub-therapeutic levels. Possible serious PSZ drug-drug interactions, seizures, in adult hematologic cancer patients should be highlighted and carefully monitored.

Background: Essential thrombocythemia (ET) is a myeloproliferative neoplasm (MPN) marked by a risk of thrombotic and hemorrhagic complications, and by a long-term risk of evolution to myelofibrosis (MF), and acute leukemia (AL). Methods: Inclusion criteria: ET diagnosed as per WHO classification 2008, age 18 to 45 year. Exclusion criteria: (hepatic and renal dysfunction, history of psychiatric disorder, in particular depression, autoimmune hepatitis, severe cardiac dysfunction, known hypersensitivity to IFN. Complete hematologic response (CR) as per ELN guidelines and molecular response (MR) was defined as 'complete' when JAK2V617F became undetectable with the technique used (i.e., V617F <1%), 'partial' when >50% decrease of baseline V617F was obtained, and 'minor' when V617F decrease was between 20% and 49%. After inclusion, Pegasys was started subcutaneously at 50 microgram weekly for 4 weeks then 135 microgramg/week from week 5. Results: Baseline characteristics of the patients are age 19-44 years, 16 males and 24 females. Five patients had a history of major thrombotic events. All the patients responded to Pegasys at the 12-month evaluation including 35 hematologic CRs (94.6%), and 5 PRs (5.4%). Cumulative incidences of hematologic CRs and PRs showed that 100% of responses were achieved within 12 months. After the first year, V617F continued to decrease without evidence of plateau, the proportion of mutant JAK2 being always similar or lower in the last sample compared with the previous one. None of the responding patients experienced an increase of V617F during follow-up. Median V617F was 58% at 12 months (P <0.001). Hematologic response was achieved in all patients within 2 months and were sustained beyond the first year. Toxicities included musculoskeletal pain in 6 patients, skin toxicity in 4, and GI symptoms in 2, none of the toxicity exceeds WHO Grade 2. Conclusion: PEGylated interferon alfa-2a is an effective and promising treatment for ET reducing the vascular risk and preventing evolution to MF, MDS, and AL. With low toxicity profiles and good hematological and molecular responses, percentage of V617F appears to be a good tool to monitor minimal residual disease and to evaluate treatment efficacy. However, further studies are needed to confirm these results.

Background and Objectives: Reference assisted assembly requires the use of a reference sequence, as a model, to assist in the assembly of novel genomes. The standard method for identifying the best reference sequence for the assembly of a novel genome aims at counting the number of reads of the novel genome that align to the reference sequences and then choosing the reference sequence which has the highest number of reads aligning to it. This work explores the use of minimum description length (MDL) principle and its two variants, the two-part MDL and sophisticated MDL, in identifying the optimal reference sequence for genome assembly. Methods: The relevance of MDL to genome assembly can be realized by understanding that genome assembly is an inference problem where the task at hand is to infer the novel genome from read data obtained from sequencing. The task of MDL is to identify the model that best describes the data and within comparative assembly framework the same meaning applies to finding the reference sequences that best describe the set of reads. This work explores the potential of three variants of MDL: two-part MDL, sophisticated MDL and minimax regret for the selection of the optimal reference sequence for comparative assembly. Results: The proposed scheme based on sophisticated MDL has been shown to work successfully for the four possible set of mutations: SNPs, insertions, inversions and deletions. The proposed scheme chooses the reference sequence which has the smaller number of SNPs, insertions and deletions. The MDL scheme is able to detect all inversions and rectify them. Conclusions: The work compared the MDL scheme with the standard method of counting the number of reads that align to the reference sequence, and found that though the standard method is a necessary condition for finding the optimal sequence, it is not the sufficient condition. Therefore, the proposed MDL scheme encompassed within itself the standard method of: counting the number of reads, by defining it in an inverted fashion as counting the number of reads that did not align to the reference sequence.

Objectives: To formulate and evaluate oral dosage forms of metformin hydrochloride (MH) having sustained-release properties that would also increase MH bioavailability and address the shortcomings in the currently marketed sustained-release tablets. Methods: MH micronized powder was dispersed in molten polymeric matrices composed of Gelucire® 50/13 and various proportions of high molecular weight hydrophilic polymers, hydrophobic oily semisolid excipients, and mucoadhesive polymeric materials. The MH loaded matrices were filled in hard gelatin capsules (HGC) each containing 500 mg MH and were subsequently characterized using differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis. The prepared HGC were subjected to content uniformity and in vitro dissolution testing according to the USP-35 compendium requirements. The dissolution data were compared to instant and sustained-release marketed tablets. The effect of incorporating various proportions of the semisolid excipients on MH dissolution release rate, were also investigated. Results: MH content of the prepared HGC ranged between 96 to 103%. All the prepared semisolid filled HGC resulted in extended-release profiles of MH that lasted between 5 to 11 hours and demonstrated a release pattern which typically follows the release from mixes of triglycerides with polyethylene glycol esters of fatty acids. The incorporation of mucoadhesive excipients like carbomer to the Gelucire® 50/13-MH semisolid matrices extended the release of MH from 5 hours initially to 9 hours as a result of the formation of a gel layer around the matrix. However, the incorporation of different hydrophilic excipients like PEG35000 and Gelucire® 44/14 along with the mucoadhesive excipients sustained the release of MH up to 11 hours. XRD analysis of the MH prepared matrices demonstrated minor changes in the crystalline nature of MH. Depending on the loading ratios and the nature of the semisolid matrices used, DSC analysis revealed the changes in MH crystallinity to be from 100 to 23%. Conclusion: HGC formulated using semisolid matrices showed promising results in extending the release of MH. However, bioavailability studies to test the ability of such Gelucire based HGC of MH to improve its bioavailability and in vivo residence times are future plans.

Background and Objectives: A pregnant female requested prenatal diagnosis for a congenital and complex eye disease segregating in her family. The three-generation pedigree of Romanian ethnic origin was suggestive of an X-linked inheritance, due to exclusively affected males and no father-to-son transmission. Affected individuals had bilateral optic nerve atrophy, microphthalmia, nystagmus, leukocoria, cataract, retinal detachment, eye tumors reported as retinoblastomas, moderate mental and motor retardation, and seizures. All efforts to obtain the detailed medical records of affected individuals were fruitless. Methods: The disease locus was mapped utilizing 78 microsatellite markers that span the X-chromosome at ~2 Mb intervals, followed by candidate genes analysis and mutations detection by Sanger sequencing. Results: Affected individuals share an ~10 Mb region between DXS1056 and GATA160B08 at Xp11.23-11.4. Candidate genes in this linkage interval included BCOR and NDP. Mutation screening identified a c.267C>A p.F89L mutation in the NPD gene in all affected individuals, previously described in a single unrelated Dutch family and speculated as causing Norrie disease. Conclusions: In retrospect, clinical symptomatology fits the Norrie disease phenotype. The reported retinoblastomas were most likely pseudogliomas characteristic of Norrie disease. Detection of the c.C267A p.F89L mutation in a second unrelated family confirms the pathogenic nature of the mutation for Norrie disease. Utilization of gene mapping through linkage analyses and candidate gene screening previously utilized exclusively for research applications, were applied at a diagnostic setting and were essential in deciphering the offending molecular defect and diagnosing a disease which due to lack of medical records and poor and misleading clinical history would have no chance of being diagnosed correctly. Clinical diagnosis and mutation identification were essential prerequisites for providing proper genetic counseling and prenatal diagnosis in this family.

Background: Metabolic syndrome is associated with the rising incidence of obesity in developed countries, particularly in urban settings and is reaching epidemic proportions. Qatar, a rapidly modernizing country, has witnessed dramatic changes in urbanization and lifestyle. Recent studies demonstrate a significant prevalence of obesity and metabolic syndrome in Qatari population. According to a New York Times article (April 2010), the International Association for the Study of Obesity has ranked Qatar sixth globally for prevalence of obesity. Epidemiological studies suggest that a poor uterine environment elicited by maternal environmental exposure may program fetus susceptibility to develop cardiovascular and metabolic disease in childhood and later in life. Several studies have demonstrated the associations between pre-pregnancy obesity, chronic hypertension and dyslipidemia, and high risk of preterm birth and intrauterine growth restriction. However, there are no studies to examine the metabolic profile of Arabian pregnant women in early pregnancy with birth outcomes. Objectives: The objective of the present proposal is to examine the association between metabolic syndrome and obesity in early pregnancy of Qatari women and the risk of delivery preterm or growth restricted singletons. Methods: Qatari mother-child cohort: a prospective cohort examining a population sample of pregnant women and their children at the prefecture of Doha, Qatar within 1 year. The pregnant mothers, at maternity clinics and hospitals of Doha, were contacted. The consented participants were invited to provide biological samples in two time points of their pregnancy. Participants were interviewed to obtain information on several environmental and dietary factors, together with anthropometric measurements and blood pressure. Biochemical analyses were performed on serum triglycerides, total cholesterol, high and low density lipoprotein cholesterol, glucose, apolipoproteins, leptin, adiponectin and Interleukins. Expected results: At the national level this will be the first mother-child cohort study in Qatar. The study shall provide evidence-based relationship between maternal occupational/environmental exposure, such as sedentary life style, obesity, and other metabolic syndromes, and the subsequent birth outcomes on the Qatari population. Conclusion: Based on the study results, the public health perspective will be addressed by policy makers at the Supreme Council of Health in Qatar.

Background and Objectives: Trauma is the leading cause of death among the young population in the Middle East, including Qatar. A significant number of trauma and injured patients will require sophisticated critical care services and a disproportionately high level of hospital resources are centered within critical care units. The introduction of an advanced ACGME structured Trauma & Critical Care (TCC) Fellowship Program at the Hamad General Hospital (HGH) in Doha, Qatar will improve patient safety and outcomes, medical care and professional satisfaction. Methods: A review of the clinical impact of the TCC Fellowship Program in the trauma service at HGH, the only tertiary and national trauma center in Qatar, was conducted after the second year of implementation to evaluate the healthcare delivery to the severely injured patients, using tools from the ICU resource, evaluation, and patient outcomes rating tool (ICU Report)®. Results: Since the implementation of the TCC Fellowship Program in 2008, structured educational and training curriculum have been implemented for physicians assigned to the trauma service at the HGH. Nine fellows have successfully completed the program and graduated. A 40% decrease of intra-hospital mortality has been observed in our trauma service. Three fellows have been promoted to consultant level and six to critical care teaching staff, enhancing staff satisfaction and the overall score of our trauma ICU. Conclusions: Advanced postgraduate medical education and training program in trauma and critical care medicine have significantly contributed to patient safety and outcome of critically injured patients in Qatar and have enhanced the overall performance of our trauma service and staff satisfaction. The fellowship program is expanding to other disciplines, and is enrolling national and international candidates in emergency medicine and anesthesia in our institution and is impacting healthcare delivery of the complex cases of injured patients in the country.

Background Consanguinity and endogamy are common in the Middle East, resulting in a higher frequency of autosomal recessive (AR) disorders. This consanguinity facilitates discovery of disease causative genes particularly after introduction of the new techniques such as Whole Exome Sequencing (WES). In order to reduce the overall socio-economic burden of such diseases, the development of diagnostic tools and prevention strategies must be a priority. To achieve these goals, the causal genes underlying human genetic diseases should be first discovered. The goal of this study is to identify novel genes causing AR diseases in Qatari population using WES. Methods Genomic DNAs were captured with the Illumina TruSeq library and sequenced with Illumina HiSeq2000. The reads were aligned to the reference genome using BWA. Variants calling and annotation were performed by SAMtools and ANNOVAR, respectively. Novel variants were defined as those having an allele frequency <0.05 in the 1000 Genomes database and predicted to be nonsynonymous substitutions. Results WES was applied on affected individuals of 3 consanguineous families with Mental Retardation (MR: 2 siblings), Peripheral Neuropathy (PN: 2 siblings) and Eye Anomaly (EA: 1 male). The mode of inheritance was assumed to be AR in MR and PN families. This led to identification of 3 and 4 homozygous candidate variants, shared by two siblings, respectively. The mode of inheritance in EA was considered to be AR and/or X-linked that led to the identification of 26 autosomal homozygous and 6 X-linked genes. The function of one of the genes on X-chromosome was related to the patient phenotype. Conclusions These preliminary interesting results highlight the power of WES, to identify potential candidate genes for AR disorders. The identified genes will be considered for functional follow-up investigation and further characterization. However, our results also highlight that the large number of population-specific variants - which may be common polymorphisms in Qatar but are so far absent from public databases - make diagnosis and discovery more difficult than in well-studied Caucasian populations. We urge genetics researchers in Qatar to actively participate in the creation of a local variant database, which will greatly facilitate such future studies.

Background and Objective: Cells exposed to stress conserve energy for the repair of cellular damage by inhibiting translational initiation. The stress stimuli can trigger several stress response pathways leading to global translational attenuation, chiefly by the phosphorylation of eIF2α and disruption of the 43S assembly, which correlates to the compartmentalization of untranslated polyadenylated mRNA in discrete cytoplasmic ribonucleoprotein complexes known as Stress Granules (SGs). In a diabetic milieu, endothelial cells (ECs) that line the lumen of the blood vessels are constantly exposed to high glucose (HG) concentration (stress), which contributes to increased oxidative stress in these cells that in turn is responsible for high rates of cardiovascular complications among diabetic individuals. However, the effect of high glucose as a stress in relation to SG assembly in ECs remains unclear. The central objective of the present study is to evaluate the role of HG-induced oxidative stress in SG assembly in ECs and whether antioxidant treatment could aid in the reversal of these effects. Methods: Mouse microvascular endothelial cells (MMECs) cultured in DMEM were exposed to normal (NG, 11mM) and high (HG, 40mM) glucose for 0-48h. DHE staining was performed to evaluate oxidative stress in ECs upon HG exposure. Fixed cells were probed for SG marker proteins (G3BP and TIAR) followed by immunofluorescence confocal microscopy. Immunoblotting was performed using protein lysate samples and was probed for p-PERK, PERK, p-eIF2α and eIF2a. N-acetyl cysteine was used as the anti-oxidant in order to study the role of oxidative stress in HG induced SG formation. Results: DHE staining indicated that oxidative stress significantly increased in HG exposed MMECs while this effect was reversed upon in NAC treatment. NAC treatment also markedly decreased HG-induced SG assembly. Conclusion: The data suggest that HG-induced oxidative stress plays a major role in SG assembly in ECs. However, more studies are warranted to evaluate the process of assembly and dis-assembly of SGs in ECs upon HG exposure, which may provide a better understanding of the role of SG formation in balancing EC apoptosis and survival in a diabetic milieu. This project was funded by QNRF-NPRP # 08-165-3-054 & 4-910-3-244.

Background & Objectives: Despite the solid foundation on epidemiological evidence and basic science research, nearly 90% of the randomized controlled trials (RCT) designed to measure the efficacy of interventions on HIV incidence failed to measure a statistically significant efficacy against HIV incidence. Here, we propose the use of computer simulations to control trials as a useful tool to overcome the difficulty of effect size estimation and outcome interpretation derived from HIV intervention RCTs. Methods: We simulated the Partners in Prevention HSV/HIV transmission study recently conducted to test the efficacy of herpes simplex virus type 2 (HSV-2) suppressive therapy by acyclovir in reducing HIV transmission. We also simulated different variations of this trial, and the Rakai male circumcision trial. We developed individual-based Monte-Carlo models parameterized by the data of these RCTs and simulated the RCTs 1000 times. To measure the efficacy of the intervention, we conducted a log-rank survival analysis for each RCT realization and estimated the statistical power as the fraction of realizations that rejected the null hypothesis. Results: Our analyses indicated that the partners in prevention RCT had only 14% likelihood to observe a statistically significant efficacy for the intervention. In contrast, a different and more potent regimen for HSV-2 suppression had 87% chance of observing a statistically significant efficacy. For the Rakai male circumcision trial simulation, 94% of the RCT realizations showed statistically significant efficacy for the intervention. Conclusions: The simulations indicate that several unexpected odds have colluded to undermine the statistical power of the partners in prevention study, and therefore it would be premature to discredit the concept of acyclovir therapy for HIV prevention based on the outcome of the partners in prevention trial. Our study highlights how in silico simulations of RCTs can provide powerful tools for optimizing the design of RCTs and predicting their outcome. Observational and biological evidence summarized in computer simulations could help us on the estimation of effect size ranges and a better understanding of the system, which might provide a powerful tool to enhance the success of any HIV intervention RCT.